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Biota<br />
Vol. 3, No. 1-2, 2002<br />
Revija za biologijo in ekologijo<br />
Journal <strong>of</strong> biology and ecology<br />
Proceedings <strong>of</strong> <strong>the</strong><br />
11th Ordinary General Meeting <strong>of</strong> Societas<br />
Euro pea Herpetologica (<strong>SEH</strong>),<br />
Zalec, Slovenija, July 13-17, 2001<br />
Drustvo za proucevanje ptic in varstvo narave<br />
Society <strong>of</strong> Bird Research and Nature Protection<br />
Drustvo varuhov okolja Radoziv<br />
Environmental Society Radoziv
Biota 3 1-2,2002<br />
11th Ordinary General Meeting <strong>of</strong> Societas<br />
Europea Herpetologica (SEN),<br />
Zalec, Slovenija, July 13-17, 2001<br />
Organised by:<br />
Environmental Society Radoziv, Zalec<br />
Organising committee:<br />
Klavdija Kac<br />
Miran Orozim<br />
Stasa Tome<br />
Milan Vogrin<br />
Gregor Vovk Petrovski<br />
Tanja Vovk Petrovski<br />
Edited by:<br />
Milan Vogrin<br />
Linguistic collaboration:<br />
Victor Kennedy<br />
Scientific committee:<br />
Dr. Wolfgang Bohme (Germany)<br />
Dr. Zoltan Korsos (Hungary)<br />
Dr. Michael R.K. Lambert (United Kingdom)<br />
Dr. Claud Miaud (France)<br />
Msc. Stasa Tome (Slovenia)<br />
Dr. Marco Zuffi (Italy)<br />
The recommended citation for particular paper is e.g.:<br />
Crnobrnja-lsailovic, J. 2003: Notes <strong>of</strong> diurnal activity in Vipera ammodytes<strong>of</strong> <strong>the</strong> central<br />
Balkans. In: Vogrin, M. (ed.). Proceedings <strong>of</strong> <strong>the</strong> 11th Ordinary General Meeting <strong>of</strong><br />
Societas Europea Herpetologica (<strong>SEH</strong>), Zalec, Slovenia, July 13-17, 2001. Biota 3 (1-2):<br />
The papers in this issue are ordered according <strong>the</strong>y alphabetical order <strong>of</strong> authors.
Vsebina/Contents<br />
3 1-2, 2OO2<br />
Milan VOGRIN<br />
Preface 7<br />
Clanki/Articles<br />
Jelka CRNOBRNJA-ISAILOVIC<br />
Notes <strong>of</strong> diurnal activity in Vipera ammodytes <strong>of</strong> <strong>the</strong> Central Balkans 9<br />
Andris CEIRANS<br />
Reptiles and amphibians <strong>of</strong> <strong>the</strong> Gauja National Park, Latvia 17<br />
Augusto GENTILLI, Stefano SCALI, Francesco BARBIERI & Franco BERNINI<br />
A three-year project for <strong>the</strong> management and <strong>the</strong> conservation <strong>of</strong> amphibians<br />
in Nor<strong>the</strong>rn Italy 27<br />
Giinter GOLLMANN, Birgit GOLLMANN, Christian BAUMGARTNER & Andrea<br />
WARINGER-LOSCHENKOHL<br />
Spawning site shifts by Rana dalmatina and Rana temporaria in response<br />
to habitat change 35<br />
Kurt GROSSENBACHER<br />
First results <strong>of</strong> a 20-year-study on Common Toad Bufo bufo in <strong>the</strong> Swiss Alps 43<br />
Daniela GUICKING, Ulrich JOGER, Michael WINK<br />
Molecular Phylogeography <strong>of</strong> <strong>the</strong> Viperine Snake Natrix maura and<br />
<strong>the</strong> Dice Snake Natrix tessellata: first results 49<br />
Adrian HAILEY & Michael R.K. LAMBERT<br />
Comparative growth patterns in Afrotropical giant tortoises (genus Geochelone) 61<br />
Miodrag JOVANOVIC, Dragana BURIC & Zoran MARKOVIC<br />
Tertiary reptiles <strong>of</strong> <strong>the</strong> central part <strong>of</strong> <strong>the</strong> Balkan peninsula 67<br />
Zoltan KORSOS & Balazs TROCSANYI<br />
Herpet<strong>of</strong>auna <strong>of</strong> Round Island, Mauritius 77<br />
Bilal KUTRUP & Nurhayat YILMAZ<br />
Preliminary data on some new specimens <strong>of</strong> Vipera barani collected from Trabzon<br />
(Nor<strong>the</strong>astern Turkey) 85<br />
A.C.AA. MEESKE, N. SCHNEEWEISS & K.J. RYBCZYNSKI<br />
Reproduction <strong>of</strong> <strong>the</strong> European Pond Turtle Emys orbicularis<br />
in <strong>the</strong> nor<strong>the</strong>rn limit <strong>of</strong> <strong>the</strong> species range 91<br />
Michele MENEGON & Sebastiano SALVIDIO<br />
Notes on habitat, egg-laying and first record <strong>of</strong> <strong>the</strong> advertising<br />
call <strong>of</strong> Hyperolius kihangensis 703<br />
Zoltan Tamas NAGY, Ulrich JOGER, Daniela GUICKING & Michael WINK<br />
Phylogeography <strong>of</strong> <strong>the</strong> European Whip Snake Coluber (Hierophis)<br />
viridiflavus as inferred from nucleotide sequences <strong>of</strong> <strong>the</strong> mitochondrial<br />
cytochrome b gene and ISSR genomic fingerprinting 109<br />
Christian PASTORELLI, Paolo LAGHI & Dino SCARAVELLI<br />
Seasonal activity and spatial distribution <strong>of</strong> a Speleomantes italicus<br />
population in a natural cave 119<br />
Christian PASTORELLI, Paolo LAGHI & Dino SCARAVELLI<br />
Speleomantes antipredator strategies: a review and new observations 127<br />
Galina V. POLYNOVA & Olga E. POLYNOVA<br />
Tail autotomy as an index <strong>of</strong> human influence on <strong>the</strong><br />
Alsophylax pipiens population in <strong>the</strong> Bogdino-Baskunchak state reserve 133
Biota 3/i-a, 2002<br />
Laura RACCA<br />
The conservation <strong>of</strong> <strong>the</strong> Agile Frog Rana dalmatina in Jersey (Channel Islands) 141<br />
Sebastiano SALVIDIO, Gabriele ALARIO, Maura Valeric PASTORINO & Mirko FERRETTI<br />
Seasonal activity and abundance <strong>of</strong> Speleomantes ambrosii in cave habitats 149<br />
Stefano SCALI & Augusto GENTILLI<br />
A comparison <strong>of</strong> main heathlands in nor<strong>the</strong>rn Italy and <strong>the</strong>ir importance<br />
for amphibian populations 155<br />
Stefano SCALI, Claudia CORTI, Augusto GENTILLI, Luca LUISELLI,<br />
Edoardo RAZZETTI & Marco A.L. ZUFFI<br />
Continental versus Mediterranean European Whip Snake<br />
Hierophis viridiflavus: a morphometric approach 161<br />
Galina. S. SUROVA<br />
The role <strong>of</strong> frog egg aggregations as a control <strong>of</strong> abiotic factors 167<br />
Judit VOROS, Zoltan KORS6S & Ferenc SZALAY<br />
A comparative morphological study <strong>of</strong> <strong>the</strong> two Hungarian<br />
discoglossid toad species Bombina spp 173<br />
Vit ZAVADIL & Arnost L. SIZLING<br />
Morphological variability in <strong>the</strong> newts <strong>of</strong> <strong>the</strong> Cristatus group 181<br />
Marco A.L. ZUFFI, Augusto GENTILLI, Edoardo RAZZETTI & Stefano SCALI<br />
Transition-hybridization areas in parapatric species<br />
<strong>of</strong> Vipera aspis group from nor<strong>the</strong>rn Italy 191<br />
Nove knjige/Book reviews<br />
Milan VOGRIN<br />
CABELA, A., GRILLITSCH, H. & TIEDEMANN (eds.) 2001:<br />
Atlas zur Verbreitung und Okologie der Amphibien und Reptilien in Osterreich 197<br />
Editor acknowledgements 198
Preface<br />
)ta 3/1-2, 2OO2<br />
The third volume <strong>of</strong> Biota is entirely devoted to herpetology. This volume includes 24 contributions<br />
presented during <strong>the</strong> 11th Ordinary General Meeting (OGM) <strong>of</strong> <strong>SEH</strong> - Societas<br />
Europaea Herpetologica.<br />
The OGM in Zalec (Slovenia) was attended by 86 participants from 24 countries who contributed<br />
37 oral presentations and 56 posters. Some <strong>of</strong> <strong>the</strong>m, as <strong>full</strong> papers, are reproduced<br />
in this volume.<br />
This volume goes to press much later than we expected. This is largely because <strong>of</strong> <strong>the</strong> sometimes<br />
extensive discussions we had (<strong>the</strong> referees and I) with <strong>the</strong> authors on how to make<br />
<strong>the</strong>ir work more comprehensible. Despite <strong>the</strong> delay, I hope that our readers (and authors as<br />
well) will find <strong>the</strong>se efforts worthwhile.<br />
I am also grateful to all who helped in different ways, both at <strong>the</strong> congress and during <strong>the</strong><br />
preparation <strong>of</strong> <strong>the</strong> <strong>proceedings</strong>. Here I wish to thank <strong>the</strong> past president <strong>of</strong> <strong>SEH</strong>, pr<strong>of</strong>. dr.<br />
Wolfgang Bohme, and <strong>the</strong> past and former secretary <strong>of</strong> <strong>SEH</strong>, dr. Michael R.K. Lambert, for<br />
<strong>the</strong>ir encouragement and work during <strong>the</strong> congress.<br />
Finally, I wish to mention that for <strong>the</strong> first time at <strong>the</strong> OGM <strong>of</strong> <strong>SEH</strong>, we started with a competition<br />
for "best student talk" and "best student poster". Congratulations Daniela and<br />
Nathalie! Daniela's paper can be found in <strong>the</strong>se <strong>proceedings</strong>.<br />
Thanks also to all referees (see <strong>the</strong> list <strong>of</strong> referees) for <strong>the</strong>ir time and help and to all participants<br />
with whom I have (almost with all) very pleasant discussions.<br />
-»»<br />
I hope that we will see you some day in Slovenia again!<br />
Milan Vogrin
CRNOBRNJA - ISAILOVIC BJOta 3/i-a. 2002 9<br />
Notes <strong>of</strong> diurnal activity in Vipera<br />
ammodytes <strong>of</strong> <strong>the</strong> Central Balkans<br />
Jelka CRNOBRNJA-ISAILOVIC<br />
Department <strong>of</strong> evolutionary biology, Institute for biological research,<br />
29. Novembra 142, 11000 Belgrade, FR Yugoslavia<br />
E-mail: jelka@ibiss.bg.ac.yu<br />
Abstract<br />
Diurnal activity in Vipera ammodytes, <strong>the</strong> most widespread viper in <strong>the</strong> Balkans, was<br />
analysed using a sample <strong>of</strong> records collected from 1981 to 2001 in Serbia and<br />
Montenegro. The inspiration for this study arose from several questions summarized as<br />
one: is it possible to success<strong>full</strong>y define activity <strong>of</strong> specimens in space and time by a<br />
combination <strong>of</strong> states <strong>of</strong> several environmental variables? A restricted number <strong>of</strong> measured<br />
environmental variables, as well as spatial/temporal heterogeneity <strong>of</strong> records,<br />
influenced <strong>the</strong> course <strong>of</strong> <strong>the</strong> study. The results pointed to significantly different diurnal<br />
activity patterns <strong>of</strong> <strong>the</strong> sexes in two seasons (spring and summer; p < 0.05). Distribution<br />
<strong>of</strong> male and female records in two seasons and three main parts <strong>of</strong> <strong>the</strong> day was also<br />
non-random. The observed differences between male and female diurnal activity patterns<br />
would be <strong>the</strong> consequence <strong>of</strong> <strong>the</strong>ir reproductive activity schedules during two seasons.<br />
Differences in diurnal activity sensu sTricto seem to be related to insolation regimes<br />
<strong>of</strong> two seasons.<br />
Key words: Vipera ammodytes, Central Balkans, diurnal activity<br />
Received 1 October 2001; accepted 4 November 2001
10 Biota 3 i-:>., ?.oo:>. CRNOBRNJA - ISAILOVIC<br />
INTRODUCTION<br />
The Balkan Peninsula comprises most <strong>of</strong><br />
<strong>the</strong> European part <strong>of</strong> <strong>the</strong> sand viper's<br />
species area (see in: Crnobrnja-lsailovic &<br />
Haxhiu 1997). In Serbia, this viper was<br />
not recorded north <strong>of</strong> <strong>the</strong> Sava and <strong>the</strong><br />
Danube river flows. Vipera ammodytes<br />
(Figure 1) is a common inhabitant <strong>of</strong><br />
canyons and gorges, as well as <strong>of</strong> areas<br />
belonging to <strong>the</strong> vegetation zone <strong>of</strong><br />
open, mostly oak xerophilous forests in<br />
Serbia and Montenegro. Syntopic reptile<br />
species in <strong>the</strong> continental part <strong>of</strong> <strong>the</strong><br />
Central Balkans frequently include:<br />
Podarcis mural/s, Lacerta viridis,<br />
Corone/la austriaca, Testudo hermanni,<br />
and in particular habitats also Lacerta<br />
pratico/a pontica, Ab/epharus kitaibelli<br />
and Coluber caspius.<br />
Figure 1. Vipera ammodytes, adult female,<br />
from Sou<strong>the</strong>astern Serbia (photo: J.<br />
Crnobrnja-lsailovic).<br />
Knowledge <strong>of</strong> <strong>the</strong> sand viper population<br />
dynamic and activity patterns in Serbia<br />
and Montenegro still does not go beyond<br />
<strong>the</strong> level <strong>of</strong> anecdotal observations. My<br />
intention in this study was to arrange diffuse<br />
data into statistically useful information<br />
and to provoke more detailed studies<br />
in <strong>the</strong> future, comparable to those<br />
already done for related species (Money<br />
1994, Zuffi 1999, Ujvari & Korsos 2000).<br />
MATERIAL AND METHODS<br />
Vipera ammodytes was recorded during<br />
<strong>the</strong> author's inventory <strong>of</strong> amphibian and<br />
reptile species in Serbia and Montenegro,<br />
from 1981 to today (Figure 2). The analyzed<br />
area broadly covers <strong>the</strong> geographic<br />
space between 42° and 44°40' North latitude.<br />
The specimens were detected within<br />
a range <strong>of</strong> altitudes 10 - 1600m above sea<br />
level. The quality <strong>of</strong> environmental data<br />
accompanying <strong>the</strong> records varied from<br />
case to case, depending on <strong>the</strong> accessibility<br />
<strong>of</strong> equipment. The terrain was checked<br />
visually during <strong>the</strong> day time by slowly<br />
walking through various habitats along <strong>the</strong><br />
planned transect route. Specimens were<br />
captured and sexed where possible. Some<br />
sand viper habitats were checked from<br />
midnight to 02h A.M., but nocturnal activity<br />
<strong>of</strong> <strong>the</strong> species was not detected.<br />
In total, 67 records were accompanied<br />
with data concerning at least one environmental<br />
variable (Table 1). Individuals were<br />
recorded during spring (1 in March, 4 in<br />
April, 14 in May and 2 in <strong>the</strong> first half <strong>of</strong><br />
June) and summer (1 in <strong>the</strong> third week <strong>of</strong><br />
June, 43 in July and 2 in August). Among<br />
<strong>the</strong>m, only mature individuals (41) were<br />
chosen for analysis. Juveniles were omitted<br />
from <strong>the</strong> study because overall sample size<br />
was too small for such a structured analysis.<br />
The sex <strong>of</strong> captured animals was recognized<br />
by presence/absence <strong>of</strong><br />
hemipenises. It is worth mentioning that<br />
15 adults were measured before being<br />
released. Their overall size varied from 404<br />
to 670 mm. The records were categorized<br />
according to accompanying variables and<br />
processed through simple (two variables)<br />
and multivariate (more than two variables)<br />
correspondence analyses (Statistica 5.0).<br />
RESULTS<br />
The results <strong>of</strong> ordinary correspondence<br />
analysis (Table 2.) point to different activity<br />
patterns <strong>of</strong> <strong>the</strong> two sexes: males were<br />
significantly more "visible" during <strong>the</strong>
CRNOBRNJA - ISAILOVIC Biota 3 1-2,2002 11<br />
Figure 2. A. European part <strong>of</strong> V. ammodytes species area. B. Location <strong>of</strong> records used in this<br />
analysis.
12 Biota 3/i-a, 2002 CRNOBRNJA - ISAILOVIC<br />
Table 1. Variables processed through various correspondence analyses.<br />
Variable<br />
Sex<br />
Season<br />
Part <strong>of</strong> <strong>the</strong> day<br />
Exposure<br />
Behaviour<br />
spring, while females were more<br />
"exposed" during <strong>the</strong> summer (Table 3).<br />
Differences in activity patterns between<br />
sexes in relation to o<strong>the</strong>r environmental<br />
variables were not detected. Also, <strong>the</strong><br />
results suggest that <strong>the</strong> activity time <strong>of</strong><br />
adult sand vipers generally differs<br />
between two seasons (p < 0.05): during<br />
<strong>the</strong> spring, <strong>the</strong> sand vipers were usually<br />
seen in <strong>the</strong> middle <strong>of</strong> <strong>the</strong> day, and during<br />
<strong>the</strong> summer, mostly in <strong>the</strong> morning hours<br />
(Table 3). The results <strong>of</strong> multiple correspondence<br />
analysis pointed to significant<br />
association <strong>of</strong> <strong>the</strong> different sexes with<br />
two seasons (spring and summer) and<br />
three main parts <strong>of</strong> <strong>the</strong> day (Table 2):<br />
males were more <strong>of</strong>ten noticed in <strong>the</strong><br />
spring middays and females in summer<br />
mornings (Figure 2). Analysis, which<br />
included sets <strong>of</strong> four as well as all five<br />
Category<br />
Male<br />
Female<br />
Spring<br />
(21stMarch-23rdJune)<br />
Summer<br />
(23rd June-20111 September)<br />
Morning<br />
(07A.M. - 1 1 A.M.)<br />
Midday<br />
(11A.M.-04P.M.)<br />
Afternoon<br />
(04PM. - 08 P.M.)<br />
South<br />
Sou<strong>the</strong>ast<br />
East<br />
Nor<strong>the</strong>ast<br />
North<br />
West<br />
Taking res tin shade<br />
Basking<br />
Moving<br />
Hiding<br />
Escaping<br />
Mating<br />
N° <strong>of</strong> cases<br />
16<br />
19<br />
21<br />
46<br />
14<br />
20<br />
7<br />
10<br />
8<br />
12<br />
1<br />
4<br />
3<br />
9<br />
8<br />
4<br />
2<br />
11<br />
2<br />
recorded variables, also showed significant<br />
non-random associations (Table 2).<br />
However, it is important to mention that<br />
no statistical significance tests are customarily<br />
applied to <strong>the</strong> results <strong>of</strong> a correspondence<br />
analysis; <strong>the</strong> primary purpose<br />
<strong>of</strong> <strong>the</strong> technique is to produce a simplified<br />
(low-dimensional) representation <strong>of</strong> <strong>the</strong><br />
information in a large frequency table.<br />
Increasing <strong>the</strong> number <strong>of</strong> variables diminishes<br />
discrimination power when <strong>the</strong><br />
sample is relatively small (Table 2). For<br />
that reason I avoided interpretation <strong>of</strong><br />
results where more than three variables<br />
were tested for associations.<br />
DISCUSSION<br />
Seasonal differences in activity patterns<br />
among sexes were observed in European<br />
populations <strong>of</strong> several viperide species
CRNOBRNJA - ISAILOVIC BJOta 3/1-2, 2002 13<br />
Table 2. A summary <strong>of</strong> simple correspondence analysis. Df = degrees <strong>of</strong> freedom; P = probability;<br />
n.s.=non-significant; *=p < 0.05; ***=p < 0.001<br />
Variables<br />
Sex X Season<br />
Sex X part <strong>of</strong> <strong>the</strong> day<br />
Sex X exposition<br />
Sex X behavior<br />
Season X part <strong>of</strong> <strong>the</strong> day<br />
Season X exposition<br />
Season X behavior<br />
part <strong>of</strong> <strong>the</strong> day X exposition<br />
Part <strong>of</strong> <strong>the</strong> day X behavior<br />
Exposition X behavior<br />
SIMPLE CORRESPONDENCE ANALYSIS<br />
X2 test value<br />
5.57<br />
5.35<br />
5.89<br />
6.68<br />
7.98<br />
9.31<br />
5.82<br />
16.05<br />
11.70<br />
26.33<br />
df<br />
1<br />
2<br />
5<br />
5<br />
2<br />
6<br />
5<br />
12<br />
10<br />
20<br />
P<br />
0.0182<br />
0.0689<br />
0.2452<br />
0.2452<br />
0.0185<br />
0.1570<br />
0.3246<br />
0.1890<br />
0.3059<br />
0.1553<br />
significance<br />
A<br />
MULTIPLE CORRESPONDENCE ANALYSIS<br />
Sex X season X part <strong>of</strong> <strong>the</strong> day<br />
Total inertia<br />
1.333<br />
Dimensions<br />
4<br />
Total X2<br />
135.345 36<br />
df<br />
P<br />
0.000<br />
significance<br />
***<br />
No. Dim. Singular<br />
value<br />
Eigenvalue %Inertia Cumulative% X2<br />
1<br />
0.751 0.564 42.26 42.26 57.20<br />
2<br />
0.613 0.376 28.21 70.48 38.18<br />
3<br />
0.462 0.214 .^ 16.03 86.50 21.69<br />
Sex X season X part <strong>of</strong> <strong>the</strong> day X exposition<br />
Total inertia<br />
2.500<br />
Dimensions<br />
10<br />
Total X2<br />
317.228<br />
df<br />
169<br />
P<br />
0.000<br />
significance<br />
***<br />
No. Dim. Singular<br />
value<br />
Eigenvalue %Inertia Cumulative% X2<br />
1<br />
0.734 0.538 21.53 21.53 68.31<br />
2<br />
0.683 0.467 18.67 40.20 59.22<br />
3<br />
0.574 0.329 13.16 53.36 41.76<br />
Sex x season X part <strong>of</strong> <strong>the</strong> day X exposition x behavior<br />
Total inertia<br />
2.600<br />
Dimensions<br />
13<br />
Total X2<br />
401.404 289<br />
Df<br />
P<br />
0.000<br />
significance<br />
***<br />
No. Dim. Singular<br />
value<br />
Eigenvalue %Inertia Cumulative% X2<br />
1<br />
0.739 0.546 21.01 21.01 84.35<br />
2<br />
0.638 0.407 15.67 36.68 62.90<br />
3<br />
0.604 0.365 14.04 50.73 56.38<br />
(recently: Biella & Volkl 1993 for Vipera<br />
berus; Bonnet & Naulleau, 1993, Saint<br />
Girons 1994, Monney 1994 and Zuffi<br />
1999 for Vipera aspis; Baron 1997 for<br />
Vipera ursinii ursinii). Generally, <strong>the</strong> earlier<br />
emergence <strong>of</strong> male vipers from hiber-<br />
n.s.<br />
n.s.<br />
n.s.<br />
*<br />
n.s.<br />
n.s.<br />
n.s.<br />
n.s.<br />
n.s.<br />
nation was explained by <strong>the</strong>ir need for<br />
completion <strong>of</strong> spermatogenesis (Nilson<br />
1980). It was also noted that pregnant<br />
females are more easily seen during <strong>the</strong><br />
summer than non-reproductive females<br />
or adult males (Andren, 1985). Having in
14 Biota 3 1-2, aooa CRNOBRNJA - ISAILOVIC<br />
Table 3. Percent <strong>of</strong> records within categories sex and time in two seasons.<br />
Sex<br />
Males<br />
Females<br />
Total<br />
part <strong>of</strong> <strong>the</strong> day<br />
Morning<br />
Midday<br />
Afternoon<br />
Total<br />
Spring<br />
75%<br />
25%<br />
100%<br />
8%<br />
64%<br />
28%<br />
100%<br />
Summer<br />
33%<br />
67%<br />
100%<br />
52%<br />
38%<br />
10%<br />
100%<br />
Figure 3. Associations <strong>of</strong> records having different categories <strong>of</strong> variables SEX, SEASON and<br />
PART OF THE DAY in <strong>the</strong> space described by <strong>the</strong> first two correspondent axes.<br />
E<br />
0<br />
1.0<br />
0.5<br />
0.0<br />
-0.5<br />
-1.0<br />
-1.5<br />
-2.0<br />
-2.5<br />
-1.5<br />
2D Plot <strong>of</strong> Column Coordinates. Dimension: 1 x 2<br />
Input Table (Rows x Columns). 7x7 (Burt Table)<br />
MALES<br />
MIDDAY<br />
AFTERNOON<br />
-1.0 -0.5 0.0 0.5<br />
mind poor quality <strong>of</strong> input data in this<br />
study (<strong>the</strong> absence <strong>of</strong> consistent longterm<br />
monitoring in <strong>the</strong> same habitat), I<br />
can only suppose that sand viper males in<br />
<strong>the</strong> Central Balkans are more active in <strong>the</strong><br />
spring searching for territory and for<br />
mates, like <strong>the</strong>ir relatives. For V.<br />
ammodytes, such behavior has been sporadically<br />
noticed in <strong>the</strong> field but still is not<br />
statistically confirmed or rejected by<br />
Dimension 1, Eigenvalue: .56349 (42.26% <strong>of</strong> Inertia)<br />
1.0 1.5<br />
appropriate experiments. Ritual male<br />
combats are also observed in this species<br />
(Steward 1971).<br />
The fact that most sand viper females<br />
were recorded in <strong>the</strong> summer logically<br />
points to <strong>the</strong>ir reproductive status: breeding<br />
in V. ammodytes in <strong>the</strong> continental<br />
part <strong>of</strong> <strong>the</strong> Central Balkans takes place in<br />
<strong>the</strong> spring (I have records <strong>of</strong> courtship at
CRNOBRNJA - ISAILOVIC Biota 3/i-a, 2002 15<br />
<strong>the</strong> end <strong>of</strong> April in 1984 in <strong>the</strong> nor<strong>the</strong>rn<br />
part <strong>of</strong> <strong>the</strong> area studied, but I have also<br />
seen courtship at <strong>the</strong> end <strong>of</strong> May 1996 in<br />
<strong>the</strong> sou<strong>the</strong>rn part); also, <strong>the</strong> female<br />
caught at <strong>the</strong> end <strong>of</strong> August 1997 at<br />
1200m above sea level had signs <strong>of</strong><br />
recent parturition. Accordingly, pregnancy<br />
<strong>of</strong> sand viper females in <strong>the</strong> study area<br />
lasts through <strong>the</strong> summer months.<br />
Parturition probably happens earlier at<br />
lower altitudes - from <strong>the</strong> beginning <strong>of</strong><br />
August in lowland areas to <strong>the</strong> end <strong>of</strong><br />
August/beginning <strong>of</strong> September in habitats<br />
near <strong>the</strong> upper altitudinal limit.<br />
Pregnant females need more energy<br />
input than males or non-reproductive<br />
females, so <strong>the</strong>y spend more time basking<br />
(Bozhanskii & Kudryavcev 1986, Andren<br />
1985 for V. berus). I collected most <strong>of</strong> <strong>the</strong><br />
summer data during July (<strong>the</strong> presumed<br />
pregnancy period) and females predominated<br />
among <strong>the</strong> specimens observed.<br />
Consequently, one can suppose that most<br />
<strong>of</strong> <strong>the</strong> sand viper females observed during<br />
<strong>the</strong> summer months in <strong>the</strong> Central<br />
Balkans were pregnant.<br />
Significant differences in time <strong>of</strong> activity<br />
among males and females <strong>of</strong> V.<br />
ammodytes in Serbia and Montenegro<br />
seem to be a direct consequence <strong>of</strong> <strong>the</strong>rmal<br />
differences between <strong>the</strong> two seasons<br />
in <strong>the</strong> temperate climatic region. A large<br />
part <strong>of</strong> <strong>the</strong> snakes' motion in space is<br />
connected with choosing <strong>the</strong> optimum<br />
insolation regime (Bozhanskii &<br />
Kudryavcev 1986). The early morning<br />
hours during <strong>the</strong> spring in this region are<br />
still too cold for helio<strong>the</strong>rmic reptile<br />
species, while summer mornings are more<br />
favorable for daily activities. On <strong>the</strong> o<strong>the</strong>r<br />
hand, midday is <strong>the</strong> most comfortable<br />
part <strong>of</strong> <strong>the</strong> day in <strong>the</strong> spring, while in <strong>the</strong><br />
summer many reptile species usually<br />
spend <strong>the</strong> warmest part <strong>of</strong> <strong>the</strong> day in<br />
<strong>the</strong>ir dens.<br />
Acknowledgements<br />
My colleagues Dr. Marco Zuffi (Pisa) and Dr Jens B. Rasmussen (Copenhagen) helped<br />
greatly with <strong>the</strong>ir suggestions about literature. I would also like to thank <strong>the</strong> anonymous<br />
reviewers for constructive comments, and Oliver Isailovic for careful preparation <strong>of</strong> illustrations.<br />
REFERENCES<br />
ANDREN, C. 1985: Risk <strong>of</strong> predation in male and female adders, Vipera berus (Linne).<br />
Amphibia-Reptilia 6: 203-206.<br />
BARON, J. - P. 1997: Demographie et dynamique d' une population francaise de Vipera<br />
ursinii ursinii (Bonaparte, 1835). These de Doctorat, Ecole Pratique des Hautes<br />
Etudes, 201 p.<br />
BIELLA, H-J. & VOLKL, W. 1993: Die Biologie der Kreuzotter (Vipera berus, L. 1758) in<br />
Mitteleuropa - ein kurzer Uberblick. Mertensiella 3: 311-318.<br />
BONNET, X. & NAULLEAU, G. 1993. Relations entre la glycemie et I' activite saisonniere<br />
chez Vipera aspis L. Amphibia-Reptilia 14: 295-306.<br />
BOZHANSKII, AT. & KUDRYAVCEV, S.V. 1986: Ecological Observations <strong>of</strong> <strong>the</strong> Rare Vipers<br />
<strong>of</strong> <strong>the</strong> Caucasus. In: Rocek, Z. (ed.) Studies in Herpetology: 495-498.<br />
CRNOBRNJA-ISAILOVIC, J. & HAXHIU, I. 1997: Vipera ammodytes. In: Case, J..-P,<br />
CABELA, A. , CRNOBRNJA-ISAILOVIC, J., DOLMEN, D., GROSSENBACHER, K.,<br />
HAFFNER, P., LESCURE, J., MARTENS, H., MARTINEZ-RICA, J.P., MAURIN, H.,<br />
OLIVEIRA, M.E., SOFIANIDOU, T.S., VEITH, M. & ZUIDERWIJK, A. (eds.) Atlas<br />
<strong>of</strong> amphibians and reptiles in Europe. <strong>SEH</strong> &MNHN (IEGP/SPN), Paris.<br />
MONNEY, J.-C. 1994: Comparaison des cycles annuels d'activite de Vipera aspis et Vipera<br />
berus (Ophidia, Viperidae) dans une station des Prealpes Bernoises (ouest de la
16 Biota 3 1-2,2002 CRNOBRNJA - ISAILOVIC<br />
Suisse). Bull.Soc.Herp. Fr. 71-72: 49-61.<br />
NILSOIM, G. 1980. Male Reproductive Cycle <strong>of</strong> European Adder, Vipera berus, and its<br />
Relation to Annual Activity Periods. Copeia 4: 729-737.<br />
SAINT-GIRONS, H. 1994: Les risques de predation lies a la reproduction chez un Viperidae<br />
ovovivipare, Vipera aspis L, d' apres les observations visuelles. Amphibia-Reptilia<br />
15:413-416.<br />
STEWARD, J.W. 1971: The snakes <strong>of</strong> Europe. Fairleigh Dickinson Univ. Press, Ru<strong>the</strong>rford-<br />
Madison-Teancek.<br />
UJVARI, B., KORSOS, Z. & PECHY, T. 2000: Life history, population characteristics and conservation<br />
<strong>of</strong> <strong>the</strong> Hungarian meadow viper (Vipera ursinii rakosiensis). Amphibia-<br />
Reptilia 21: 267-278.<br />
ZUFFI, M. A. L. 1999: Activity patterns in a viviparous snake, Vipera aspis (L.), from<br />
Mediterranean central Italy. Amphibia-Reptilia 20: 313-318.
CEIRANS Biota 3/1-2.2002 17<br />
Reptiles and amphibians <strong>of</strong> <strong>the</strong><br />
Gauja National Park, Latvia<br />
Andris CEIRANS<br />
Department <strong>of</strong> Zoology and Animal Ecology, Faculty <strong>of</strong> Biology, University <strong>of</strong> Latvia<br />
Kronvalda bulv. 4, Riga LV-1586, Latvia<br />
E-mail: andrisc@lanet.lv<br />
Abstract<br />
An inventory <strong>of</strong> <strong>the</strong> herpet<strong>of</strong>auna <strong>of</strong> <strong>the</strong> Gauja National Park, located in <strong>the</strong> north-central<br />
part <strong>of</strong> Latvia, was carried out in 1999-2000. Its objectives were to determine<br />
species composition, status, and habitat preferences. The main attention was focussed<br />
on reptiles. Data were collected along transects located throughout <strong>the</strong> territory <strong>of</strong> <strong>the</strong><br />
Park. The total length <strong>of</strong> transects was 166.2 km, and numerous separate observations<br />
<strong>of</strong> various species were also recorded. Common and widespread species were Lacerta<br />
vivipara, Bufo bufo, Rana temporaria, and Rana synklepton esculenta. Anguis fragilis<br />
was found mostly in a dry pine, pine-spruce forest on <strong>the</strong> terrace <strong>of</strong> <strong>the</strong> ancient valley<br />
<strong>of</strong> Gauja River. A large population <strong>of</strong> Matrix natrix was found in <strong>the</strong> sou<strong>the</strong>rn part <strong>of</strong> <strong>the</strong><br />
Park in deciduous and coniferous forests. A few populations <strong>of</strong> Lacerta agiliswere found<br />
in dry pine forests, and on <strong>the</strong> banks <strong>of</strong> <strong>the</strong> Gauja River. Rana atvalis was a rare species,<br />
more frequently found in high moors. There were also several records <strong>of</strong> Triturus cristatus<br />
and T. vulgaris in <strong>the</strong> Gauja National Park. The required conservation activities are<br />
discussed.<br />
Key words: reptiles, amphibians, habitats, Gauja National Park, Latvia<br />
Received 6 September 2001; accepted 28 October 2001
ta 3/1-2, 2OO2 CEIRANS<br />
INTRODUCTION<br />
Gauja National Park is located in <strong>the</strong><br />
north-central part <strong>of</strong> Latvia, about 35 km<br />
north-west <strong>of</strong> RTga. The Park was established<br />
in 1973, and it was <strong>the</strong>n <strong>the</strong> second<br />
National Park in <strong>the</strong> territory <strong>of</strong> <strong>the</strong><br />
former USSR. The area <strong>of</strong> <strong>the</strong> Park is<br />
91,745 ha; forest occupies 48,592 ha<br />
(Pilats 2000). The area is dissected by <strong>the</strong><br />
ancient valley <strong>of</strong> <strong>the</strong> Gauja River, toge<strong>the</strong>r<br />
with valleys <strong>of</strong> its numerous tributaries<br />
and side ravines. Gauja National Park is<br />
one <strong>of</strong> <strong>the</strong> florally richest regions in<br />
Latvia, with a high diversity <strong>of</strong> forests.<br />
The largest proportion <strong>of</strong> old broadleaved<br />
forests in Latvia is found near <strong>the</strong><br />
town <strong>of</strong> Sigulda (Pilats 2000). However,<br />
boreal coniferous forests dominate <strong>the</strong><br />
Park. The largest high moor is Sudas-<br />
Zviedru mire (2575 ha), found in <strong>the</strong><br />
sou<strong>the</strong>rn part <strong>of</strong> Gauja National Park.<br />
There are also several smaller mires in <strong>the</strong><br />
central and nor<strong>the</strong>rn parts <strong>of</strong> <strong>the</strong> Park.<br />
Agricultural landscapes, although occupying<br />
a large part <strong>of</strong> <strong>the</strong> Park, are fragmented<br />
by many forest stands, shrubs,<br />
and fallow lands that serve as refuges for<br />
wild animals. Gauja National Park also<br />
includes many historically significant<br />
objects and popular tourist sites.<br />
The present survey is part <strong>of</strong> a large-scale<br />
inventory <strong>of</strong> flora, fauna and habitats <strong>of</strong><br />
Gauja National Park carried out in 1999<br />
and 2000. The aim <strong>of</strong> <strong>the</strong> inventory <strong>of</strong><br />
herpet<strong>of</strong>auna was to determine <strong>the</strong><br />
species composition and status <strong>of</strong> reptile<br />
and amphibian species, and to describe<br />
<strong>the</strong>ir habitats in <strong>the</strong> Park. The main<br />
attention was focussed on reptiles,<br />
because studies <strong>of</strong> <strong>the</strong>se animals in<br />
Latvia have been very few and since<br />
information on <strong>the</strong>ir preferred habitats is<br />
limited.<br />
METHODS<br />
Data was collected mainly on transects in<br />
<strong>the</strong> field seasons <strong>of</strong> 1999 (from 06.08. to<br />
08.09.) and 2000 (from 17.04. to<br />
14.07). The total length <strong>of</strong> transects was<br />
166.2 km. Censuses were carried out in<br />
all <strong>of</strong> <strong>the</strong> main habitat groups <strong>of</strong> <strong>the</strong> Park<br />
(Table 1). The type <strong>of</strong> habitat was determined<br />
from short descriptions made in<br />
<strong>the</strong> field. This information was supplemented<br />
by data from forest plans <strong>of</strong> <strong>the</strong><br />
State Forest Service. The syntaxonomical<br />
classification <strong>of</strong> Latvian forests has not<br />
yet been <strong>full</strong>y developed (Prieditis 1999),<br />
and is <strong>the</strong>refore supplemented (Table 1)<br />
by <strong>the</strong> forest stand classification applied<br />
in <strong>the</strong> forestry industry (Buss 1997). The<br />
transects were evenly distributed<br />
throughout <strong>the</strong> area. Therefore, <strong>the</strong><br />
transect length for a particular habitat is<br />
roughly proportional to <strong>the</strong> area that <strong>the</strong><br />
habitat occupies in <strong>the</strong> Park. An exception<br />
was agricultural landscapes, which<br />
were considerably less represented in<br />
transects (31.0 % <strong>of</strong> total transect length<br />
in comparison to 45 % coverage in <strong>the</strong><br />
Park).<br />
The routes were carried out in warm and<br />
dry wea<strong>the</strong>r, as <strong>the</strong> main attention was<br />
focussed on reptiles. Each observation <strong>of</strong><br />
'a reptile or amphibian was mapped at a<br />
1:50.000 scale. For every reptile specimen<br />
observed, a brief description <strong>of</strong> <strong>the</strong><br />
site was made, and in most cases it was<br />
supplemented later with information<br />
from <strong>the</strong> data base <strong>of</strong> <strong>the</strong> State Forest<br />
Service. Transects were located mostly<br />
along sites with potentially highest reptile<br />
density (roadsides, fringes, cuttings,<br />
clearings etc.), and extrapolation <strong>of</strong> this<br />
data to <strong>the</strong> whole habitat could provide<br />
misleading results. Densities were used<br />
only for Anguis fragilis and Lacerta vivipara<br />
for comparing various habitats. To<br />
describe <strong>the</strong> occurrence <strong>of</strong> <strong>the</strong> Common<br />
Lizard Lacerta vivipara in habitats, two<br />
parameters were used: <strong>the</strong> number <strong>of</strong><br />
<strong>the</strong> individuals and <strong>the</strong> number <strong>of</strong><br />
records. As <strong>the</strong> movement range for<br />
Lacerta vivipara is up to 80 m<br />
(Zarnolodchikov & Avilova 1989), two<br />
records were considered to be separate if
CEIRANS 3/1-2, 2OO2 19<br />
Table 1. Transect length <strong>of</strong> different habitats <strong>of</strong> <strong>the</strong> Gauja National Park<br />
Plant communities after Prieditis 1999* and Kabucis 2000**, forest types after Buds 1997.<br />
Habitat<br />
Dry pine forests on poor sandy<br />
soil<br />
Dry mesotrophic pine and pinespruce<br />
forests<br />
Dry mesotrophic spruce forests<br />
Moist eutrophic broad-leaved<br />
forests<br />
High moors with pine and pine<br />
forests on wet peat<br />
Degraded high moors and pine<br />
forests on drained peat<br />
O<strong>the</strong>r wet forest types<br />
Edges <strong>of</strong> dry pine, pine-spruce<br />
forests<br />
Edges <strong>of</strong> dry spruce and leaf<br />
tree forests<br />
Agricultural landscapes,<br />
meadows and shrubs<br />
Total<br />
Plant community<br />
Cladonio-Pinetum,Vaccinio<br />
vitis-idaeae-Pinetum*<br />
Vaccinio myrtilli-Pinetum,<br />
several types not classified<br />
yet*<br />
Oxalido-Piceetum exeelsae*<br />
Querco-Tilietum*<br />
Sphagnion<br />
magellanici**,Vaccinio<br />
uliginosi-Pinetum*<br />
not classified yet*<br />
<strong>the</strong> distance between individuals was<br />
more than 100 meters. This served to<br />
reduce <strong>the</strong> effect <strong>of</strong> occasional observa- ~~*l<br />
tions <strong>of</strong> a large number <strong>of</strong> individuals at<br />
<strong>the</strong> same site due to higher activity in<br />
optimal wea<strong>the</strong>r conditions or better visibility<br />
for <strong>the</strong> observer. Juveniles were<br />
excluded from <strong>the</strong>se data. In <strong>the</strong> analysis<br />
<strong>of</strong> dominant tree species in habitats, data<br />
acquired from transect censuses were<br />
supplemented with descriptions <strong>of</strong><br />
twelve locations where <strong>the</strong> reptile<br />
species were observed outside transects.<br />
For amphibians, <strong>the</strong> type <strong>of</strong> habitat was<br />
determined later from State Forest<br />
Service forest plans and its data base.<br />
Amphibians along transects were not<br />
counted, and densities were not calculated<br />
in cases when numerous individuals<br />
were observed. The transect method was<br />
not applied for <strong>the</strong> recording <strong>of</strong> newts,<br />
and information on <strong>the</strong>se animals was<br />
collected occasionally.<br />
Species distribution maps were prepared<br />
using <strong>the</strong> Baltic Co-ordinate System,<br />
Forest type<br />
Cladinoso-callunosa,<br />
Vaccinosa<br />
Myrtillosa,<br />
Hylocomiosa<br />
Oxalidosa<br />
Aegopodiosa<br />
moor, Sphagnosa,<br />
Caricoso-phragmitosa<br />
Callunosa turf, me).,<br />
Vaccinosa turf. mel.<br />
Myrtilloso-sphagnosa,<br />
Dryopterioso-caricosa<br />
Myrtillosa,<br />
Hylocomiosa<br />
Oxalidosa,<br />
Aegopodiosa<br />
Transect length,<br />
km (%)<br />
2.8(1.7)<br />
78.1 (47.0)<br />
9.3 (5.6)<br />
2.3(1.4)<br />
8.4(5.1)<br />
1.7(1.0)<br />
1.1 (0.7)<br />
7.5 (4.5)<br />
3.4(2.0)<br />
51.6(31.0)<br />
166.2 (100.0)<br />
Transverse Mercator Projection (TM-<br />
1993). The number <strong>of</strong> 1x1 km squares <strong>of</strong><br />
this Co-ordinate System, in which<br />
species were observed, was used to estimate<br />
<strong>the</strong> occurrence <strong>of</strong> various species in<br />
<strong>the</strong> Gauja National Park. A total <strong>of</strong> 269<br />
or 27.5 % <strong>of</strong> all 1x1 km squares <strong>of</strong> <strong>the</strong><br />
Park were visited in <strong>the</strong> survey. The<br />
number <strong>of</strong> <strong>the</strong> squares is not necessary<br />
identical to <strong>the</strong> number <strong>of</strong> locations indicated<br />
for species in <strong>the</strong> results, as several<br />
locations in <strong>the</strong> same square are possible.<br />
RESULTS<br />
Six reptile and five amphibian species<br />
were found during <strong>the</strong> survey. At least<br />
one species was observed in 201 or 74.7<br />
% <strong>of</strong> <strong>the</strong> visited 1x1 km squares.<br />
Observation frequency for <strong>the</strong> various<br />
species is shown in Table 2.<br />
Reptiles<br />
The Sand Lizard Lacerta agilis was found<br />
in 2 areas: dry pine Pinus sylvestris forest
20 3/1-2, 2OO2 CEIRANS<br />
Table 2. Occurrence <strong>of</strong> reptiles and amphibians in 1x1 km squares <strong>of</strong> <strong>the</strong> Baltic Co-ordinate<br />
System that were crossed by transects in <strong>the</strong> Gauja National Park<br />
Species<br />
Reptiles<br />
Lacerta agilis<br />
Lacerta vivipara<br />
Anguis fragilis<br />
Natrix natrix<br />
Vipera bents<br />
Amphibians<br />
Triturus cristatus<br />
Triturus vulgaris<br />
Bufo bufo<br />
Rana arvalis<br />
Rana temporaria<br />
Rana synklepton esculenta<br />
in <strong>the</strong> south-western part <strong>of</strong> <strong>the</strong> Park (2<br />
locations), and banks and terraces <strong>of</strong> <strong>the</strong><br />
river Gauja and smaller tributaries in <strong>the</strong><br />
central part <strong>of</strong> <strong>the</strong> Park (5 locations).<br />
The habitats <strong>of</strong> Lacerta agilis can be<br />
grouped in two types:<br />
1. low and sparse Pinus sylvestris growing<br />
(sometimes mixed with birch Betula<br />
spp. and spruce Picea abies) on dry sand,<br />
with a tree height <strong>of</strong> 2-7 m and canopy<br />
cover <strong>of</strong> 10-30 %; herb layer dominated<br />
by grasses (Poa, Festuca, Calamagrostis)<br />
and, in some cases, with horsetail<br />
(Equisetum); herb cover <strong>of</strong> 10-50%;<br />
habitat found on <strong>the</strong> banks <strong>of</strong> <strong>the</strong> river<br />
and in some disturbed habitats (such as<br />
old sand quarries) on terrace (5 locations);<br />
2. fringes <strong>of</strong> dry and tall pine forest<br />
(Vaccinio vitis-idaeae - Pinetum,<br />
Vacdnio myrtilli - Pinetum associations)<br />
on sandy soil on terrace (2 locations).<br />
The Common Lizard Lacerta vivipara was<br />
more or less evenly distributed throughout<br />
<strong>the</strong> territory <strong>of</strong> <strong>the</strong> Gauja National<br />
No. <strong>of</strong><br />
squares<br />
6<br />
59<br />
3<br />
9<br />
1<br />
1<br />
1<br />
91<br />
8<br />
133<br />
40<br />
% <strong>of</strong> visited<br />
squares<br />
2.2<br />
21.9<br />
3.0<br />
3.3<br />
0.4<br />
0.4<br />
0.4<br />
33.8<br />
3.0<br />
49.4<br />
14.9<br />
Park. This species was observed in most<br />
<strong>of</strong> <strong>the</strong> habitats, except some types <strong>of</strong><br />
forest. The highest density was in high<br />
""moors and wet pine forests on peat,<br />
especially in drained sites, and along <strong>the</strong><br />
fringes <strong>of</strong> various dry forests (Table 3).<br />
The dominant tree species in Lacerta<br />
vivipara wet forest habitats usually was<br />
pine Pinus sylvestris, while deciduous<br />
trees (Betula spp., Populus tremula,<br />
Quercus robur, Tilia cordata) were characteristic<br />
<strong>of</strong> open habitats with separate<br />
trees or small groups <strong>of</strong> trees, such as in<br />
open agricultural landscapes.<br />
Lacerta vivipara preferred forest habitats,<br />
especially mature dry forest, where it<br />
was observed exclusively on relatively<br />
open sites such as forest clearcuts, grassy<br />
roadsides, banks, fringes and sites with<br />
large gaps in <strong>the</strong> forest canopy.<br />
The Slow Worm Anguis fragilis was<br />
found rarely, in upland pine dominated<br />
forest areas in <strong>the</strong> whole territory, and<br />
only in dry forest types. Specimens were<br />
usually observed on or near paths. Two
CEIRANS 3/1-2, 2OO2 21<br />
Table 3. Occurrence <strong>of</strong> Lacerta vivipara on transects in various habitats.<br />
Only <strong>the</strong> habitats where species were found are included in <strong>the</strong> list. The first value indicates<br />
<strong>the</strong> number <strong>of</strong> observations, <strong>the</strong> second - density <strong>of</strong> findings. For <strong>the</strong> transect<br />
length in <strong>the</strong> habitat see Table 1. Separate observations that were made outside <strong>the</strong><br />
transects are not included.<br />
Habitat<br />
Dry mesotrophic pine and pine-spruce forests<br />
High moors with pine and pine forests on wet peat<br />
Degraded high moors and pine forests on drained peat<br />
Fringes <strong>of</strong> dry pine, pine-spruce forests<br />
Fringes <strong>of</strong> dry spruce and leaf tree forests<br />
Agricultural landscapes, meadows and shrubs<br />
locations (or 0.71 records/km) were in<br />
pine forest on poor sandy soil (association<br />
Vaccinia vitis-idaeae - Pinetum),<br />
and seven locations (0.09 records/km) in<br />
pine and pine-spruce forest on<br />
mesotrophic soil. Pinus sylvestris was <strong>the</strong><br />
dominant tree species in all locations.<br />
The Grass Snake Natrix natrix was regularly<br />
observed in <strong>the</strong> south-western part<br />
<strong>of</strong> <strong>the</strong> Park, particularly near Sigulda.<br />
The species inhabited <strong>the</strong> following habitats:<br />
young and patchy Alnus incana and<br />
Betula stands and meadows with pools<br />
located on <strong>the</strong> bed <strong>of</strong> <strong>the</strong> ancient Gauja<br />
river valley (5 locations), old broadleaved<br />
forests with Fraxinus excelsior,<br />
Quercus robur, and Ulmus glabra on <strong>the</strong><br />
slopes <strong>of</strong> <strong>the</strong> valley (association Querco-<br />
Tilietum, 2 locations), and dry<br />
mesotrophic pine and pine-spruce<br />
forests on terraces <strong>of</strong> <strong>the</strong> valley (4 locations).<br />
In forest habitats, it was usually<br />
found in habitat edges and roadsides.<br />
There were also some records <strong>of</strong> Natrix<br />
natrix in 1985-1987 from o<strong>the</strong>r parts <strong>of</strong><br />
<strong>the</strong> Park, mostly along <strong>the</strong> Gauja River<br />
(Z. Bruneniece, unpublished Bachelor's<br />
<strong>the</strong>sis).<br />
The Adder Vipera berus was found in<br />
only two locations during <strong>the</strong> survey. The<br />
first was on <strong>the</strong> edge <strong>of</strong> a clay quarry<br />
with young and sparse Betula and Picea<br />
abies, with Calluna vulgaris in <strong>the</strong><br />
ground cover. This site was located on a<br />
Records<br />
12/0.15<br />
5/0.60<br />
3/1.79<br />
5/0.67<br />
2/0.59<br />
15/0.29<br />
Individuals<br />
15/0.19<br />
5/0.60<br />
8/4.71<br />
5/0.67<br />
2/0.59<br />
17/0.33<br />
slope with western exposure, within dry<br />
mesotrophic pine and spruce forests in<br />
<strong>the</strong> nor<strong>the</strong>rn part <strong>of</strong> <strong>the</strong> Park. The o<strong>the</strong>r<br />
observation <strong>of</strong> Vipera berus was made<br />
about 500 m outside <strong>the</strong> eastern border<br />
<strong>of</strong> <strong>the</strong> Park, on <strong>the</strong> grassy railway<br />
embankment near a young Betula-Salix-<br />
Alnus incana stand in a hilly landscape.<br />
Vipera berus is also present in <strong>the</strong> nor<strong>the</strong>astern<br />
part <strong>of</strong> Sudas-Zviedru mire<br />
(observed by M. Deicmane); however,<br />
<strong>the</strong> species was not detected <strong>the</strong>re during<br />
<strong>the</strong> transect counts.<br />
Amphibians<br />
Two species were common on transects,<br />
and were found in various habitats<br />
throughout <strong>the</strong> entire territory <strong>of</strong> <strong>the</strong><br />
Park - <strong>the</strong> Common Toad Bufo bufo and<br />
<strong>the</strong> Common Frog Rana temporaria.<br />
Rana temporaria was <strong>the</strong> dominant<br />
species in all natural habitats, except for<br />
<strong>the</strong> Sudas-Zviedru high moor and sometimes<br />
also <strong>the</strong> broad-leaved forests on<br />
<strong>the</strong> slopes <strong>of</strong> <strong>the</strong> Gauja River valley. Bufo<br />
bufo were fewer in number than Rana<br />
temporaria in all dry forest habitats, with<br />
<strong>the</strong> exception <strong>of</strong> a few cases in broadleaved<br />
forest on valley slopes (association<br />
Querco-Tilietum). This species was<br />
infrequent on high moors and in wet<br />
pine forests on peat. Both Bufo bufo and<br />
Rana temporaria were relatively common<br />
on agricultural landscapes, especial-
22 Biota 3/1-2,2002 CEIRANS<br />
ly Bufo bufo, which was a usual inhabitant<br />
<strong>of</strong> <strong>the</strong> local villages. Both species<br />
were absent in <strong>the</strong> driest fragments <strong>of</strong><br />
pine forests on sandy soil (association<br />
Cladinio-Pinetum).<br />
Green frogs Rana synklepton esculenta<br />
were common in aquatic and semi-aquatic<br />
habitats in forest and agricultural landscapes,<br />
especially wet habitats with many<br />
pools. Green frogs were usual inhabitants<br />
<strong>of</strong> ponds on agricultural farms. Most <strong>of</strong><br />
<strong>the</strong> examined individuals belonged to <strong>the</strong><br />
Pool Frog Rana lessonae; a few specimens<br />
<strong>of</strong> <strong>the</strong> Green Frog Rana esculenta<br />
were found in <strong>the</strong> largest lake <strong>of</strong> <strong>the</strong> Park<br />
(Ungura Lake, 394 ha).<br />
The Moor Frog Rana arvalis was a rare<br />
species; a few individuals were occasionally<br />
observed in <strong>the</strong> entire territory <strong>of</strong> <strong>the</strong><br />
Park. It was more common in <strong>the</strong> central<br />
parts <strong>of</strong> <strong>the</strong> Sudas-Zviedru mire, <strong>the</strong><br />
largest high moor <strong>of</strong> <strong>the</strong> Gauja National<br />
Park, and in <strong>the</strong> surrounding wet forest<br />
where it was <strong>the</strong> dominant amphibian<br />
species.<br />
Amphibian roadkills are fairly common in<br />
<strong>the</strong> Gauja National Park, especially <strong>of</strong><br />
Bufo bufo. The number <strong>of</strong> killed individuals<br />
<strong>of</strong> this species on country roads during<br />
spring migration in one case<br />
(26.04.2000) reached 43 toads on a<br />
0.025 km long road span, and in two<br />
o<strong>the</strong>r cases (17.04.2000 and<br />
17.05.2000), <strong>the</strong>re were 11 and 12 killed<br />
toads per 0.035 and 1.5 km, respectively.<br />
Separate killed individuals <strong>of</strong> this species<br />
were a common sight in or near human<br />
settlements. Brown frogs were seldom<br />
killed on roads, but on one day<br />
(17.05.2000), 11 dead frogs per km were<br />
recorded.<br />
Two newt species were found at one<br />
location each. A dead specimen <strong>of</strong> <strong>the</strong><br />
Great Crested Newt Triturus cristatus was<br />
found on a road on <strong>the</strong> outskirts <strong>of</strong> <strong>the</strong><br />
town <strong>of</strong> Cesis. The road is located<br />
between a wet deciduous tree forest area<br />
and a garden area. There have been also<br />
reports from zoologists (V. Pilats, A.<br />
Minde) about findings in four o<strong>the</strong>r locations<br />
in ponds near or in human settlements.<br />
Larvae <strong>of</strong> <strong>the</strong> Smooth Newt<br />
Triturus vulgaris were found in <strong>the</strong> central<br />
part <strong>of</strong> <strong>the</strong> Park in a small forest lake with<br />
swampy banks surrounded by wet pine<br />
forest. There are two reports (from M.<br />
Deicmane, M. Kalnins) about findings in<br />
human settlements.<br />
Comments on some o<strong>the</strong>r species<br />
In 1988, a few individuals <strong>of</strong> <strong>the</strong> Fire-bellied<br />
Toad Bombina bombina were<br />
observed (by S. Inberga) near Sigulda in a<br />
popular tourist area, but <strong>the</strong> species has<br />
not survived in this location. Apparently<br />
<strong>the</strong> observed individuals were released by<br />
man. Bombina bombina in Latvia is on<br />
<strong>the</strong> periphery <strong>of</strong> <strong>the</strong> distribution, and <strong>the</strong><br />
species inhabits only <strong>the</strong> sou<strong>the</strong>rn part <strong>of</strong><br />
Latvia - Bauska and Daugavpils Districts<br />
(unpublished data).<br />
The Natterjack Toad Bufo calamita was<br />
found (by M. Kalnins) in <strong>the</strong> 1990s in two<br />
locations within a few kilometres <strong>of</strong> <strong>the</strong><br />
"Sou<strong>the</strong>rn border <strong>of</strong> <strong>the</strong> Gauja National<br />
Park, and <strong>the</strong> presence <strong>of</strong> this species in<br />
<strong>the</strong> Park is highly possible.<br />
There are records <strong>of</strong> <strong>the</strong> European Pond<br />
Turtle Emys orbicularis in 1914 and 1925<br />
in <strong>the</strong> territory <strong>of</strong> <strong>the</strong> present-day Gauja<br />
National Park (Silins, Lamsters 1934).<br />
Since <strong>the</strong>n, <strong>the</strong>re have been no new<br />
records <strong>of</strong> this species, in spite <strong>of</strong> well<br />
developed tourism in <strong>the</strong> Park. In Latvia,<br />
<strong>the</strong>re have been rare but regular findings<br />
<strong>of</strong> Emys orbicularis in various regions. In<br />
most cases, <strong>the</strong>se animals were possibly<br />
released by man, but in a few locations in<br />
sou<strong>the</strong>rn Latvia (Daugavpils and Dobele<br />
. districts) this species is observed regularly<br />
(personal communications by E. Tone, M.<br />
Pupins) and small populations <strong>of</strong> this<br />
species are expected <strong>the</strong>re.
CEIRANS Biota 3/i-2, 2OO2 23<br />
DISCUSSION<br />
A survey using methods similar to those<br />
<strong>of</strong> <strong>the</strong> present study was carried out in<br />
1994-1997 in <strong>the</strong> Kemeri National Park<br />
(Ceirans, in press). That National Park is<br />
located in <strong>the</strong> central part <strong>of</strong> Latvia, but<br />
only 15 % <strong>of</strong> its territory is covered by<br />
agricultural land and human settlements<br />
(compared to 45 % in <strong>the</strong> Gauja National<br />
Park). The frequency <strong>of</strong> records (% <strong>of</strong> visited<br />
1x1 km squares where species was<br />
found) for Lacerta vivipara, L. agilis and<br />
Vipera berus was about <strong>the</strong> same in both<br />
Parks. The frequency for Anguis fragilis<br />
and Matrix natrix records was respectively<br />
about 3 and 5 times higher in <strong>the</strong><br />
Kemeri National Park. For <strong>the</strong> latter<br />
species, this difference is probably associated<br />
with climatic differences.<br />
The large proportion <strong>of</strong> agricultural landscapes<br />
in <strong>the</strong> Gauja National Park affects<br />
to a great extent <strong>the</strong> status <strong>of</strong> species.<br />
Four species <strong>of</strong> reptiles and amphibians<br />
(Lacerta vivipara, Bufo bufo, Rana ternporaria,<br />
and Rana synklepton esculenta)<br />
are common and widespread in <strong>the</strong><br />
whole territory. All <strong>of</strong> <strong>the</strong>se species are*»<br />
typical <strong>of</strong> agricultural landscapes <strong>of</strong> <strong>the</strong><br />
Park, where farming methods are not as<br />
intensive as in Western Europe or in some<br />
o<strong>the</strong>r Latvian regions. The remaining<br />
species are not adapted to <strong>the</strong>se landscapes<br />
(Anguis fragilis, Vipera berus,<br />
Triturus vulgaris, Triturus cristatus) or<br />
(and) are near <strong>the</strong> limits <strong>of</strong> <strong>the</strong>ir climatic<br />
tolerance (Lacerta agilis, Natrix natrix),<br />
which are <strong>the</strong> reasons for <strong>the</strong>ir rarity.<br />
Rana arvalis was found in <strong>the</strong> same habitats<br />
as Rana temporaria, but usually in<br />
considerably lower numbers (excepting<br />
for some high moor areas). Among frogs<br />
killed on roads in spring 1999-2000 in <strong>the</strong><br />
sou<strong>the</strong>rn part <strong>of</strong> <strong>the</strong> Gauja National Park,<br />
not far from <strong>the</strong> Sudas-Zviedru high moor<br />
(I. Valikova, unpublished Bachelor's <strong>the</strong>sis),<br />
Rana arvalis and R. temporaria individuals<br />
were found in proportions from<br />
1:5 to 1:10.<br />
The Adder Vipera berus population is<br />
probably declining in <strong>the</strong> Gauja National<br />
Park. In a survey carried out in 1985-<br />
1987, and based on verified reports from<br />
local residents, (Z. Bruneniece, unpublished<br />
Bachelor's <strong>the</strong>sis), records <strong>of</strong><br />
Anguis fragilis, Natrix natrix, and Vipera<br />
berus were present in a proportion<br />
1:0.9:0.6, compared to a proportion <strong>of</strong><br />
0.6:1:0.05 in <strong>the</strong> present survey (1999-<br />
2000) based on transect routes. As <strong>the</strong><br />
total number <strong>of</strong> findings <strong>of</strong> <strong>the</strong>se three<br />
species was considerably higher in <strong>the</strong><br />
former survey, a decrease <strong>of</strong> all three<br />
species is possible. Populations <strong>of</strong> Anguis<br />
fragilis, Natrix natrix, and Vipera berus<br />
show long-term declining trends in<br />
Finland (Terhivuo 1993), and decline <strong>of</strong><br />
Vipera berus is recorded for North<br />
European Russia (Orlov & Ananjeva<br />
1995). Long-term declining trends for<br />
<strong>the</strong>se species in Latvia is also possible.<br />
Lacerta vivipara inhabits diverse habitats<br />
in both forest and open landscapes <strong>of</strong> <strong>the</strong><br />
Park. For this species, a change <strong>of</strong> dominant<br />
tree species depending on humidity<br />
and canopy closure in <strong>the</strong> habitat was<br />
observed. There was a correlation<br />
between <strong>the</strong> occurrence <strong>of</strong> Lacerta vivipara<br />
and that <strong>of</strong> pine in wet and closed<br />
habitats, and that <strong>of</strong> deciduous trees in<br />
dry open habitats. The latter possibly<br />
reflects <strong>the</strong> dominance <strong>of</strong> deciduous trees<br />
in open landscapes <strong>of</strong> <strong>the</strong> Park, and not<br />
that deciduous trees were preferred by<br />
Lacerta vivipara. However, <strong>the</strong>se shifts<br />
can reflect <strong>the</strong> microclimatic preferences<br />
<strong>of</strong> <strong>the</strong> species. Pine stands are usually well<br />
lit, and in <strong>the</strong> case <strong>of</strong> wet stands (such as<br />
in high moor or forest on wet peat), fairly<br />
open. Deciduous tree stands in wet<br />
sites are usually shaded and may be too<br />
cool for Lacerta vivipara. In <strong>the</strong> open<br />
landscapes, summer temperatures can be<br />
considerably higher than in <strong>the</strong> forest<br />
sites. Sparse, dry and relatively open pine<br />
stands may be too dry and hot for Lacerta<br />
vivipara, and suitable ra<strong>the</strong>r to Lacerta<br />
agilis which inhabits such sites on <strong>the</strong>
24 Biota 3/1-2,2002 CEIRANS<br />
n high moors <strong>of</strong> <strong>the</strong> Moscow region <strong>of</strong><br />
Russia, <strong>the</strong> most favourable habitat for<br />
Lacerta vivipara was observed to be wet<br />
sites with sparse low pine stands and<br />
Myrica gale, Ledum palustre,<br />
Eriophorum vaginatum, and Sphagnum<br />
in <strong>the</strong> herb and moss layer<br />
(Zamolodchikov & Avilova 1989). In<br />
high moors <strong>of</strong> <strong>the</strong> Gauja National Park,<br />
Lacerta vivipara preferred sites with a<br />
stable water regime, such as sparse low<br />
pine-birch stands near old harvested<br />
peat areas or in partially drained moor<br />
areas, with characteristic swards <strong>of</strong><br />
Calluna vulgaris and grasses (Molinia<br />
caerulea etc.) in <strong>the</strong> herb layer.<br />
During <strong>the</strong> survey, large numbers <strong>of</strong><br />
killed amphibians on roads were<br />
observed in relatively few cases, mostly<br />
regarding Bufo bufo. There are data<br />
from <strong>the</strong> sou<strong>the</strong>rn part <strong>of</strong> <strong>the</strong> Park, near<br />
Ligatne (I.Valikova, unpublished<br />
Bachelors <strong>the</strong>sis), where amphibians<br />
killed on roads were counted during <strong>the</strong><br />
spawning period (April - first 10 days <strong>of</strong><br />
May) for two seasons (1999-2000). The<br />
maximum numbers <strong>of</strong> kills were 212<br />
Rana temporaria, 35 Bufo bufo, and 14<br />
Rana an/alls in one spring per 1 km <strong>of</strong><br />
road. In one case, about 15 km outside<br />
<strong>the</strong> border <strong>of</strong> <strong>the</strong> Gauja National Park,<br />
382 killed Rana temporaria and 70 Bufo<br />
bufo per km were counted. This indicates<br />
that <strong>the</strong> number <strong>of</strong> killed brown<br />
frogs recorded in <strong>the</strong> Park in <strong>the</strong> present<br />
survey - maximum 11 frogs/km - is an<br />
REFERENCES<br />
underestimate, probably due to <strong>the</strong> timing<br />
<strong>of</strong> field work after <strong>the</strong> end <strong>of</strong> <strong>the</strong><br />
amphibian spawning season.<br />
The following conservation and research<br />
activities are recommended for <strong>the</strong> Gauja<br />
National Park:<br />
- Creating a data base <strong>of</strong> records at least<br />
for rare species (Lacerta agilis, Anguis<br />
fragilis, Natrix natrix, Vipera berus,<br />
Triturus vulgaris, T. cristatus, Rana<br />
an/alls) using <strong>the</strong> 1999-2000 inventory<br />
project as a base;<br />
- More research on ecology <strong>of</strong> all species<br />
is needed to encourage <strong>the</strong> development<br />
<strong>of</strong> a management plan for reptiles and<br />
amphibians in <strong>the</strong> Gauja National Park;<br />
- Development <strong>of</strong> a network <strong>of</strong> protected<br />
microreserves for Triturus cristatus.<br />
The species is included in Appendices II<br />
<strong>of</strong> both <strong>the</strong> Bern Convention<br />
(Anonymous 1992a) and ED Directive<br />
92/43EEC (Anonymous 1992b), and in<br />
<strong>the</strong> 2nd category (vulnerable species) <strong>of</strong><br />
<strong>the</strong> Red Data Book <strong>of</strong> Latvia (Ingelog et.<br />
al. 1993);<br />
- Measures should be made to avoid<br />
"•amphibian road kills. In cases <strong>of</strong> mass<br />
road kills, conservation activities such as<br />
construction <strong>of</strong> roadside fences and<br />
underground passages should be<br />
planned;<br />
- Increasing public awareness <strong>of</strong> reptile<br />
and amphibian conservation in <strong>the</strong> Park.<br />
Fur<strong>the</strong>r tourism development could have<br />
a negative effect on some species, particularly<br />
snakes.<br />
ANONYMOUS 1992a: Appendix II, Strictly Protected Fauna Species to <strong>the</strong> Convention on<br />
<strong>the</strong> Conservation <strong>of</strong> European Wildlife and Natural Habitats, Bern, 1979.<br />
Directorate <strong>of</strong> Environment and Local Authorities, Strasbourg.<br />
ANONYMOUS 1992b: EU Directive 92/43/EEC on <strong>the</strong> Conservation <strong>of</strong> Natural Habitats and<br />
Wild Fauna and Flora, Brussels.<br />
BUSS, K. 1997: Forest ecosystem classification in Latvia. Proceedings <strong>of</strong> <strong>the</strong> Latvian Academy<br />
<strong>of</strong> Sciences. Section B 51: 204-218.<br />
CEIRANS, A. in press: Reptiles and anurans <strong>of</strong> <strong>the</strong> Kemeri National Park, Latvia. In: Heikkila,
CEIRANS Biota 3/i-a, 2002 25<br />
R. & Lindholm, T. (eds.). Biodiversity and conservation <strong>of</strong> <strong>the</strong> boreal nature.<br />
Proceedings <strong>of</strong> <strong>the</strong> 10 years anniversary symposium <strong>of</strong> <strong>the</strong> Nature Reserve<br />
Friendship. The Finnish Environment 485.<br />
INGELOG, T., ANDERSSON, R. & TJERNBERG, M. (eds.) 1993: Red Data Book <strong>of</strong> <strong>the</strong> Baltic<br />
Region. Part 1. Lists <strong>of</strong> Threatened Vascular Plants and Vertebrates. Swedish<br />
Threatened Species Unit, Uppsala.<br />
KABUCIS, I. (ed.) 2000: Biotopu rokasgramata. Eiropas Savienibas aizsargajamie biotopi<br />
Latvija [Hand<strong>book</strong> <strong>of</strong> habitats. Protected habitats by European Union in Latvia].<br />
Latvijas dabas fonds, Riga (in Latvian).<br />
ORLOV, N.L. & ANANJEVA, N.B. 1995: Distribution <strong>of</strong> amphibians and reptiles and <strong>the</strong>ir<br />
relict populations in <strong>the</strong> Gulf <strong>of</strong> Finland and Lake Ladoga. Memoranda Societatis<br />
pro Fauna et Flora Fennica 71: 109-112.<br />
PILATS, V. 2000: Emeralds <strong>of</strong> Latvia. Opportunities for nature tourism. Ministry <strong>of</strong><br />
Environmental Protection and Regional Development, Riga.<br />
PRIEDITIS, N. 1999: Latvian forest: nature and diversity. WWF, Riga.<br />
SILINS, J. & LAMSTERS, V. 1934: Latvijas rapuli un abinieki [Reptiles and amphibians <strong>of</strong><br />
Latvia]. Valters un Rapa, Riga (in Latvian).<br />
TERHIVUO, J. 1993: Provisional atlas and status <strong>of</strong> populations for <strong>the</strong> herpet<strong>of</strong>auna <strong>of</strong><br />
Finland in 1980-92. Annales Zoologici Fennici 30: 55-69.<br />
ZAMOLODCHIKOV, D.G. & AVILOVA, K.V. 1989: Materials on biology <strong>of</strong> <strong>the</strong> Common<br />
Lizard, Lacerta vivipara, in a high moor in Western Moscow region. In:<br />
Amphibians and reptiles <strong>of</strong> Moscow Region. Nauka, Moscow: 147-152 (in<br />
Russian).
GENTILLI, SCALI, BARBIERI & BERNINI Biota 3/i-a, 2002 27<br />
A three-year project for <strong>the</strong><br />
management and <strong>the</strong> conservation<br />
<strong>of</strong> amphibians in Nor<strong>the</strong>rn Italy<br />
Augusto GENTILLI1, Stefano SCALI2r<br />
Francesco BARBIERI1 & Franco BERNINI1<br />
1 Dip. Biologia Animale, University <strong>of</strong> Pavia, p.za Botta 9, 27100 Pavia, Italy<br />
2 Museo Civico di Storia Naturale, c.so Venezia 55, 20121 Milano-ltaly<br />
Abstract<br />
In 1998 <strong>the</strong> Lombardy District (Regione Lombardia, Nor<strong>the</strong>rn Italy), in co-operation<br />
with <strong>the</strong> Italian Ministry <strong>of</strong> Environment, started an integrated three-year project for<br />
conservation <strong>of</strong> amphibians in twelve regional parks, in both mountain and plain areas.<br />
The aims <strong>of</strong> <strong>the</strong> project were translocation <strong>of</strong> some threatened amphibian species and<br />
habitat management <strong>of</strong> some endangered environments. The target species were<br />
Salamandra salamandra, Triturus camifex, Pelobates fuscus insubricus, Bombina variegata,<br />
Hyla intermedia, Rana latastei, R. dalmatina and R. temporaria. The target habitats<br />
are natural and artificial ponds and springs located in areas where <strong>the</strong>se environments<br />
are disappearing.<br />
The translocations involved P. fuscus insubricus and R. latastei; eggs <strong>of</strong> <strong>the</strong>se species<br />
were collected in areas in Nor<strong>the</strong>rn Italy in accordance with taxonomic and conservation<br />
interests. Eggs and tadpoles were bred in semi-natural conditions by <strong>the</strong> biologists<br />
<strong>of</strong> <strong>the</strong> University <strong>of</strong> Pavia. Tadpoles were released immediately before metamorphosis.<br />
One half <strong>of</strong> bred tadpoles were released in <strong>the</strong> areas where <strong>the</strong> eggs were collected.<br />
The new and restored ponds in most cases have been spontaneously colonized by seven<br />
amphibian species: Salamandra salamandra, Bufo bufo, Hyla intermedia, Rana dalmatina,<br />
R. latastei, R. temporaria and R. synklepton esculenta. The successful metamorphosis<br />
and survival after <strong>the</strong> wintering period <strong>of</strong> <strong>the</strong> translocated specimens were also<br />
observed. These preliminary results underline <strong>the</strong> importance and <strong>the</strong> validity <strong>of</strong> <strong>the</strong>se<br />
methods <strong>of</strong> translocation and habitat management for amphibian conservation projects.<br />
Key words: Amphibians, conservation, translocation, habitat management, Nor<strong>the</strong>rn<br />
Italy<br />
Received 24 September; accepted 12 December 2001
28 Biota 3/1-2,2002 GENTILLI, SCALI, BARBIERI & BERNINI<br />
INTRODUCTION<br />
The use <strong>of</strong> habitat management and<br />
translocations for amphibian conservation<br />
has been proposed by many authors<br />
(Fog 1988, Burke 1991, Dodd & Seigel<br />
1991, Reinert 1991, Bray 1994, Langton<br />
et al. 1994, Amtkjaer 1995, Andren &<br />
Nilson 1995a, b, Cooke & Oldham 1995,<br />
Gubbels 1995, Hels & Fog 1995, Juul<br />
1995, Zvirgzds et al. 1995, Griffiths,<br />
1996, Agger 1997, Briggs 1997, Fog<br />
1997, Jensen 1997, Lacoste & Durrer<br />
1998). A three year project based on<br />
<strong>the</strong>se methods was started in 1998 to<br />
preserve amphibian species and <strong>the</strong>ir<br />
habitats in Lombardy (Nor<strong>the</strong>rn Italy).<br />
The project was made possible thanks to<br />
<strong>the</strong> appropriation <strong>of</strong> € 250.000 by <strong>the</strong><br />
Regione Lombardia and <strong>the</strong> Italian<br />
Ministry <strong>of</strong> <strong>the</strong> Environment. The<br />
authors describe <strong>the</strong> methods used during<br />
<strong>the</strong> project and its preliminary results.<br />
MATERIALS AND METHODS<br />
The project involved 12 natural parks,<br />
seven <strong>of</strong> which are located in <strong>the</strong> Po<br />
Plain (Table 1) and five in <strong>the</strong> Alps (Table<br />
2). The present or historical presence <strong>of</strong><br />
amphibian species in <strong>the</strong> parks was<br />
assessed by bibliographic and personal<br />
data (Andreone et al. 1993, Societas<br />
Herpetologica Italica 1996, Bonini et al.<br />
2000).<br />
Using <strong>the</strong>se data, we identified target<br />
species based on two main criteria:<br />
endemic or rare taxa considered important<br />
by European laws (Habitats<br />
Directive 92/43/European Community:<br />
T. carnifex, P. fuscus insubricus, B. variegata,<br />
H. intermedia, R. dalmatina, R.<br />
latastei) and species that, although not<br />
rare in Nor<strong>the</strong>rn Italy, are in decline in<br />
some areas (i.e., S. salamandra, H. intermedia,<br />
R. dalmatina and R. temporaria).<br />
The literature on <strong>the</strong>se species was<br />
examined to ga<strong>the</strong>r data on <strong>the</strong>ir ecological<br />
needs. Only Pelobates fuscus insubricus<br />
and Rana latastei were chosen for<br />
translocations, because <strong>the</strong>y are quite<br />
rare in many areas <strong>of</strong> Lombardy and <strong>the</strong>y<br />
are endemic to <strong>the</strong> Po Plain.<br />
Forty suitable areas for habitat management<br />
and translocations were identified<br />
following many field surveys with park<br />
staff. The causes <strong>of</strong> amphibian decline in<br />
<strong>the</strong>se sites were analysed and some<br />
Actions for habitat restoration were proposed,<br />
in accordance with <strong>the</strong> guidelines<br />
<strong>of</strong> <strong>the</strong> I.U.C.N., <strong>of</strong> <strong>the</strong> National Wildlife<br />
Institute (I.N.F.S.), and <strong>of</strong> <strong>the</strong> Societas<br />
Table 1. Target species and conservation measures chosen for lowland parks; HM<br />
Habitat Management, Rl = Reproduction, PR = Population reinforcement.<br />
Park<br />
Parco Pineta di Appiano<br />
Gentile e Tradate<br />
Parco Ticino<br />
Parco Agricolo Sud<br />
Parco Adda Sud<br />
Parco Serio<br />
Parco Oglio Sud<br />
Parco Mincio<br />
PLAIN PARKS<br />
Target species<br />
Salamandra salamandra<br />
Triturus carnifex<br />
Rana dalmatina<br />
Pelobates fuscus<br />
Pelobates fuscus<br />
Rana latastei<br />
Pelobates fuscus<br />
Rana latastei<br />
Rana latastei<br />
Rana latastei<br />
Rana latastei<br />
HM<br />
Conservation<br />
measures<br />
HM,RI<br />
HM,RI<br />
HM,RI<br />
HM,RI<br />
HM,PR<br />
HM,RI<br />
HM<br />
HM
GENTILLI, SCALI, BARBIERI & BERNINI Biota 3/1-2,2002 29<br />
Table 2. Target species and conservation measures chosen for alpine parks; HM =<br />
Habitat Management, Rl = Reintroduction, PR = Population reinforcement.<br />
Park<br />
Parco Monte Barro<br />
Parco Orobie Valtel lines!<br />
Parco Colli di Bergamo<br />
Parco Adamello<br />
Parco Alto Garda Bresciano<br />
Herpetologica Italica (I.N.F.S. 1995,<br />
Stanley Price & Fairclough 1997, Societas<br />
Herpetologica Italica 1997). In particular,<br />
we chose public areas that guaranteed<br />
major protection status over private<br />
ones; <strong>the</strong> sites where habitat restoration<br />
was not possible were discarded. The<br />
choice <strong>of</strong> sites was also based on ecological,<br />
habitat structural and human factors<br />
(Scali et al. 2002). In addition, <strong>the</strong> chosen<br />
sites were contiguous with o<strong>the</strong>r<br />
suitable damp areas, in order to re-create<br />
metapopulations.<br />
Many techniques were used for habitat<br />
restoration:<br />
• creation <strong>of</strong> new ponds: some pools<br />
were excavated in suitable areas to<br />
increase reproductive sites;<br />
• restoration <strong>of</strong> old ponds: some desiccated<br />
or almost desiccated existing<br />
ponds were excavated to increase water<br />
levels during <strong>the</strong> breeding period;<br />
• waterpro<strong>of</strong>ing <strong>of</strong> ponds: new and old<br />
ponds were waterpro<strong>of</strong>ed with clay or<br />
PVC, if necessary, to guarantee water<br />
permanence;<br />
• increase <strong>of</strong> laying points: some deadwood<br />
was introduced to facilitate egg<br />
anchorage;<br />
• excavation <strong>of</strong> tributary canals: when<br />
necessary, <strong>the</strong>y were dug to allow water<br />
permanence;<br />
• positioning <strong>of</strong> metallic grids: <strong>the</strong>se were<br />
used to stop predator fish entrance;<br />
• removal <strong>of</strong> fish: <strong>the</strong>se vertebrates were<br />
removed, if already present, using an<br />
ALPINE PARKS<br />
Target species<br />
Salamandra salamandra<br />
Rana temporaria<br />
Bombina variegata<br />
Rana temporaria<br />
Salamandra salamandra<br />
Hyla intermedia<br />
Rana temporaria<br />
Conservation measures<br />
HM<br />
HM<br />
HM<br />
HM,PR<br />
HM<br />
electrostunner, and <strong>the</strong>y were released in<br />
contiguous damp areas.<br />
Translocations were carried on only after<br />
habitat management and only if we considered<br />
reintroductions or population<br />
reinforcements necessary: i) when a population<br />
was extinct and re-colonization<br />
was impossible or unlikely, ii) when a<br />
population had strongly declined and its<br />
<strong>full</strong> recovery was unlikely or it required a<br />
very long time.<br />
The eggs <strong>of</strong> Pelobates fuscus insubricus<br />
and Rana latastei were collected in sites<br />
where <strong>the</strong>se species are quite abundant<br />
and located near <strong>the</strong> translocation areas,<br />
to guarantee genetic homogeneity. In<br />
fact, no geographic barriers are present<br />
in areas involved in <strong>the</strong> plan, and <strong>the</strong>re<br />
are no borders to <strong>the</strong> biogeographic<br />
regions. Fur<strong>the</strong>rmore, no information<br />
about <strong>the</strong> genetics <strong>of</strong> Rana latastei and<br />
Pelobates fuscus insubricus are available<br />
for <strong>the</strong>se areas, and no funds were<br />
appropriated for this kind <strong>of</strong> research.<br />
Eggs were transported to Bosco Negri<br />
Natural Reserve, which is managed by<br />
<strong>the</strong> Italian League for Bird Protection<br />
(LIPU), and <strong>the</strong>y were hatched in seminatural<br />
conditions by <strong>the</strong> authors. Eggs<br />
coming from different clumps were<br />
mixed to increase genetic diversity.<br />
Tadpoles were released only when <strong>the</strong>ir<br />
hind legs were developing; one half <strong>of</strong><br />
<strong>the</strong>m were released in <strong>the</strong> donor sites,<br />
because we do not want to impoverish<br />
those populations.
30 Biota 3/i-a, 2002 GENTILLI, SCALI, BARBIERI & BERNINI<br />
RESULTS<br />
To date, 56 habitat management projects<br />
have been carried out in 34 different<br />
sites. During <strong>the</strong> year 2000 a total <strong>of</strong><br />
2000 tadpoles <strong>of</strong> P. fuscus and 12,000 <strong>of</strong><br />
R. latastei were bred; in 2001, <strong>the</strong> number<br />
<strong>of</strong> R. latastei increased to 28,000. In<br />
<strong>the</strong> same year P. fuscus did not breed,<br />
due to unfavourable climatic conditions.<br />
In <strong>the</strong> former year we made six reintroductions<br />
<strong>of</strong> P. fuscus insubricus, and two<br />
reintroductions and three population<br />
reinforcements <strong>of</strong> R. latastei. During <strong>the</strong><br />
second year we made seven reintroductions<br />
and six population reinforcements<br />
<strong>of</strong> R. latastei.<br />
Metamorphosis occurred without any<br />
particular problem in both years and<br />
some juveniles <strong>of</strong> <strong>the</strong> spadefoot toad<br />
were found in spring 2001. The habitat<br />
management works led to an improvement<br />
<strong>of</strong> environmental conditions for<br />
amphibians in general. In fact, <strong>the</strong> populations<br />
<strong>of</strong> different species already living<br />
in those areas <strong>of</strong>ten colonized <strong>the</strong> new<br />
ponds and <strong>the</strong> number <strong>of</strong> egg clumps<br />
increased in <strong>the</strong> second year (Table 3 and<br />
DISCUSSION<br />
Preliminary data suggest that <strong>the</strong> project<br />
is quite successful. This result is apparent<br />
from <strong>the</strong> increase <strong>of</strong> breeding specimens<br />
<strong>of</strong> <strong>the</strong> species already existing in managed<br />
areas and by <strong>the</strong> successful metamorphosis<br />
<strong>of</strong> many tadpoles <strong>of</strong> <strong>the</strong><br />
translocated species. No information is<br />
available at <strong>the</strong> present time about<br />
future reproductive success <strong>of</strong> <strong>the</strong> managed<br />
populations. To achieve <strong>the</strong>se data,<br />
a monitoring phase is required in future<br />
years. The breeding operations, <strong>the</strong><br />
translocations, and <strong>the</strong> habitat management<br />
works will continue through 2002<br />
to improve <strong>the</strong> preliminary results.<br />
This kind <strong>of</strong> project guarantees a fast and<br />
easy way to reconstitute wild populations;<br />
in fact, amphibians can be bred at<br />
a low cost and a large number <strong>of</strong> future<br />
breeders can be achieved in a short time;<br />
moreover, <strong>the</strong> breeding and <strong>the</strong> release<br />
do not involve behavioural problems, as<br />
is <strong>of</strong>ten reported for birds and mammals<br />
(Bloxam & Tonge 1995).<br />
It is important to underline that many <strong>of</strong><br />
The increased species are protected by<br />
European laws (Habitat Directive). Our<br />
observations highlight <strong>the</strong> importance <strong>of</strong><br />
a co-ordinate project for an homogeneous<br />
line <strong>of</strong> conduct and for an optimization<br />
<strong>of</strong> economic resources.
GENTILLI, SCAU, BARBIERI & BERNINI Biota 3/i-a, 2002 31<br />
Table 3. Increase <strong>of</strong> already present species and colonization by new species in <strong>the</strong> lowland<br />
parks.<br />
Plain parks<br />
Parco Pineta di Appiano Gentile<br />
e Tradate<br />
Parco Ticino<br />
Parco Agricolo Sud<br />
Parco Adda Sud<br />
Parco Serio<br />
Parco Oglio Sud<br />
Parco Mincio<br />
Increase<br />
T. vulgaris<br />
T. camifex<br />
R dalmatina<br />
R. synklepton esculenta<br />
R. latastei<br />
R. latastei<br />
Colonization<br />
S. salamandra<br />
T. carnifex<br />
R. dalmatina<br />
T. vulgaris<br />
B. bufo<br />
H. intermedia<br />
R. latastei<br />
R. dalmatina<br />
R. synklepton esculenta<br />
H. intermedia<br />
R. synklepton esculenta<br />
H. intermedia<br />
R. synklepton esculenta<br />
R. synklepton esculenta<br />
R. synklepton esculenta<br />
R. synklepton esculenta<br />
Table 4. Increase <strong>of</strong> already present species and colonization by new species in <strong>the</strong><br />
alpine parks.<br />
Plain parks Increase Colonization<br />
Parco Monte Barro<br />
Parco Orobie Valtellinesi<br />
Parco Colli di Bergamo<br />
Parco Adamello<br />
Parco Alto Garda Bresciano<br />
"*& salamandra<br />
R. temporaria<br />
S. salamandra<br />
B. variegata<br />
R. temporaria<br />
S. salamandra R, temporaria<br />
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COILMANN, COLLMANN, BAUMGARTNER & WARINGER BJQta 3/1-2, 2002 35<br />
Spawning site shifts by Rana<br />
dalmatina and Rana temporaria in<br />
response to habitat change<br />
Gunter GOLLMANN1, Birgit GOLLMANN1,<br />
Christian BAUMGARTNER2 & Andrea<br />
WARINGER-LOSCHENKOHL2<br />
'Institut fur Zoologie, Universitat Wien, Althanstr. 14, A-1090 Wien, Austria<br />
E-mail: guenter.gollmann@univie.ac.at<br />
2lnstitut fur Okologie und Naturschutz, Universitat Wien, Althanstr. 14, A-1090 Wien,<br />
Austria<br />
Abstract<br />
Between 1995 and 2001 we investigated spatial patterns <strong>of</strong> spawn deposition by Rana<br />
dalmatina and R. temporaria in <strong>the</strong> flood retention basin <strong>of</strong> <strong>the</strong> Mauerbach stream in<br />
western Vienna (Austria). This basin was constructed in <strong>the</strong> late 19th century; until<br />
1996 it was bypassed by <strong>the</strong> stream flowing in a straight channel and contained several<br />
temporary pools. In <strong>the</strong> course <strong>of</strong> reconstruction works <strong>the</strong> basin was flooded for prolonged<br />
periods in 1997 and 1998 and a meadow close to <strong>the</strong> stream, outside <strong>the</strong> retention<br />
basin, was inundated. Since 1998 <strong>the</strong> stream has been running partly through <strong>the</strong><br />
basin, connecting all <strong>the</strong> pools that existed previously. Owing to <strong>the</strong> rise <strong>of</strong> <strong>the</strong> water<br />
table, a few new shallow pools formed in <strong>the</strong> upper part <strong>of</strong> <strong>the</strong> basin. Since 2000,<br />
beaver dams have stabilized <strong>the</strong> water level in <strong>the</strong> lower part <strong>of</strong> <strong>the</strong> basin. Clutches <strong>of</strong><br />
R. dalmatina were usually laid separately, attached to vegetation such as stalks <strong>of</strong> reed,<br />
whereas egg masses <strong>of</strong> R. temporaria were <strong>of</strong>ten aggregated in clusters. Both species<br />
readily adopted new water bodies and ceased to use some areas frequented in <strong>the</strong> early<br />
years <strong>of</strong> <strong>the</strong> study. A shift to shallower spawning sites, which was more distinct in R.<br />
temporaria than in R. dalmatina, may be interpreted as predator avoidance or a preference<br />
for warm microhabitats under generally colder conditions.<br />
Key words: amphibians, ecology, oviposition, stream restoration, Ranidae<br />
Received 10 November; accepted 15 December 2001
36 Biota 3/i-a, 2002 GOLLMANN, GOLLMANN, BAUMGARTNER & WARINGER<br />
INTRODUCTION<br />
Spawn is an important stage in <strong>the</strong> lifehistory<br />
<strong>of</strong> most amphibians. Selection <strong>of</strong><br />
oviposition sites by spawning frogs has<br />
consequences for <strong>the</strong> risks <strong>of</strong> desiccation<br />
and predation on embryos and larvae as<br />
well as for growth opportunities <strong>of</strong> <strong>the</strong><br />
tadpoles.<br />
Two species <strong>of</strong> brown frogs, Rana dalmatina<br />
and Rana temporaria, have widely<br />
overlapping distributions in Europe<br />
(Gasc et al. 1997), but syntopic occurrence<br />
is rare in many regions. Their lifehistories<br />
are similar; in Central Europe<br />
both species breed in early spring. The<br />
question whe<strong>the</strong>r <strong>the</strong>ir distributions are<br />
influenced by interspecific competition<br />
has remained unresolved (e.g. Riis 1988,<br />
Rohrbach & Kuhn 1997). Competition<br />
among amphibians is expected to occur<br />
mainly during <strong>the</strong> larval stage (Wilbur<br />
1996). Hence, spatial and temporal patterns<br />
<strong>of</strong> spawn deposition determine <strong>the</strong><br />
potential for competition among tadpoles,<br />
and may also influence <strong>the</strong> outcome<br />
<strong>of</strong> this competition.<br />
To assess possibilities for competition<br />
and conditions <strong>of</strong> co-occurrence, we<br />
have investigated oviposition, embryonic<br />
and larval development in syntopic populations<br />
<strong>of</strong> R. dalmatina and R. temporaria<br />
at <strong>the</strong> western outskirts <strong>of</strong> Vienna<br />
Figure 1. Water bodies in <strong>the</strong> study area 1995-1997. Outlines show maximal size <strong>of</strong><br />
temporary pools that mostly dried during summer.
GOLLMANN, GOLLMANN, BAUMGARTNER & WARINGER Biota 3/1-2, 2002 37<br />
since 1995 (Baumgartner et al. 1996,<br />
Gollmann et al. 1999). Reconstruction<br />
works affecting <strong>the</strong> study area, including<br />
a stream restoration project, caused pr<strong>of</strong>ound<br />
habitat change. Here, we describe<br />
<strong>the</strong> spatial patterns <strong>of</strong> spawn deposition<br />
over seven study years.<br />
STUDY AREA AND HABITAT CHANGE<br />
Observations were made in <strong>the</strong> flood<br />
retention basin <strong>of</strong> <strong>the</strong> Mauerbach stream.<br />
This basin was constructed in <strong>the</strong> late<br />
19th century to retard water during flood<br />
peaks. It measures approximately 350 x<br />
70 m and until 1997 was bypassed by <strong>the</strong><br />
stream flowing in a straight channel.<br />
Rgure 2. Water bodies in <strong>the</strong> study area since 1998.<br />
Pool dry in 2001<br />
Beaver dam<br />
Temporary pools in <strong>the</strong> central and lower<br />
parts <strong>of</strong> <strong>the</strong> basin usually dried in late<br />
spring or early summer (Figure 1). In<br />
1995, water from <strong>the</strong> stream never<br />
flowed into <strong>the</strong> basin. On 8 April 1996<br />
melting <strong>of</strong> snow in upstream areas caused<br />
a brief flooding <strong>of</strong> <strong>the</strong> basin, which led to<br />
<strong>the</strong> formation <strong>of</strong> a new ephemeral pool at<br />
<strong>the</strong> upper end <strong>of</strong> <strong>the</strong> basin.<br />
In autumn 1996 reconstruction works<br />
started, with <strong>the</strong> aims <strong>of</strong> improving flood<br />
protection (<strong>of</strong> downstream districts <strong>of</strong><br />
Vienna) and redirecting part <strong>of</strong> <strong>the</strong><br />
stream through <strong>the</strong> retention basin. In<br />
spring <strong>of</strong> 1997, <strong>the</strong> basin was flooded by<br />
<strong>the</strong> stream for prolonged periods, and a
38 Biota 3/1-2, 2002 GOLLWIANN, GOLLMANN, BAUAAGARTNER & WARINGER<br />
meadow close to <strong>the</strong> stream, directly<br />
above <strong>the</strong> retention basin, was inundated<br />
(Figure 1). Heavy rainfalls in July<br />
1997 caused deep submersion <strong>of</strong> <strong>the</strong><br />
retention basin, which lasted until<br />
February 1998, when debris from <strong>the</strong><br />
outlet was removed. Since <strong>the</strong>n, <strong>the</strong><br />
stream has been running partly through<br />
<strong>the</strong> basin, connecting all ponds that had<br />
existed previously. Owing to <strong>the</strong> rise <strong>of</strong><br />
<strong>the</strong> water table, a few new shallow<br />
ponds formed in <strong>the</strong> upper parts <strong>of</strong> <strong>the</strong><br />
basin. Since winter 1999/2000, beavers<br />
have inhabited <strong>the</strong> basin; <strong>the</strong>y have<br />
gradually been building dams, which<br />
have increased and stabilized <strong>the</strong> water<br />
level in <strong>the</strong> retention basin (Figure 2).<br />
MATERIAL AND METHODS<br />
During <strong>the</strong> brown frog breeding season<br />
<strong>the</strong> study area was usually visited daily,<br />
rarely at two day intervals. Minimummaximum<br />
<strong>the</strong>rmometers and watergauges<br />
were placed at several sites in <strong>the</strong><br />
study area.<br />
Each egg mass was individually marked<br />
Figure 3. Partition <strong>of</strong> <strong>the</strong> study area<br />
into four sections (see text and<br />
Figure 4).<br />
with a floating disk <strong>of</strong> cork tied to <strong>the</strong><br />
jelly with a string (1995-1999) or with a<br />
straw pushed through <strong>the</strong> center <strong>of</strong> <strong>the</strong><br />
spawn clump (2000-2001). Position <strong>of</strong><br />
<strong>the</strong> clutch, water depth at <strong>the</strong> spawning<br />
site and depth <strong>of</strong> <strong>the</strong> water column<br />
above <strong>the</strong> spawn clump were recorded;<br />
one egg was removed for species determination<br />
by enzyme electrophoresis<br />
(Baumgartner et al. 1996), which was<br />
performed for most clutches.<br />
RESULTS<br />
To summarize spatial patterns <strong>of</strong> spawn<br />
deposition, <strong>the</strong> study area is here divided<br />
into four sections (Figures 3, 4): partitions<br />
<strong>of</strong> <strong>the</strong> basin correspond to slight<br />
rises in <strong>the</strong> ground, whereas pools outside<br />
<strong>the</strong> basin are summarized as area D.<br />
In <strong>the</strong> first study year, only pools in <strong>the</strong><br />
lower and central parts <strong>of</strong> <strong>the</strong> basin (A,<br />
B) were available as breeding habitats.<br />
Newly created water bodies, such as <strong>the</strong><br />
meadow inundated in 1997 (area D),<br />
were readily accepted for spawning by<br />
both species.<br />
Figure 4. Number <strong>of</strong> clutches <strong>of</strong> R. dalmatina<br />
and R. temporaria per year in <strong>the</strong> four parts <strong>of</strong><br />
<strong>the</strong> study area (see Figure 3).<br />
1995 1996 1997 1998 1999 2000 2001<br />
Rana dalmatina H Rana temporaria
GOLLMANN, GOLLMANN, BAUMGARTNER & WARINCER 3/1-2, 2002 39<br />
Figure 5. Water depth at <strong>the</strong> spawning sites: Lower symbols indicate depth <strong>of</strong> <strong>the</strong> water<br />
body at <strong>the</strong> spawning site (mean and standard deviation) for each species and year,<br />
upper symbols show mean values for depth <strong>of</strong> <strong>the</strong> water column above <strong>the</strong> spawn<br />
clump.<br />
A •*•»<br />
ffl 20-<br />
30~<br />
40 -1<br />
A Sana dalmatina<br />
4» Sana temporaria<br />
\s <strong>of</strong><br />
1995 1996 1997 1998 1399 2000 2001<br />
always deposited singly, <strong>of</strong>ten attached<br />
to a central axis provided by a stalk <strong>of</strong><br />
reed or a branch. In <strong>the</strong> first four study<br />
years, <strong>the</strong> earliest clutches were deposited<br />
at <strong>the</strong> same site (<strong>the</strong> easternmost pool<br />
in area B, Figure 3). When <strong>the</strong> water<br />
level in <strong>the</strong> basin increased due to flooding<br />
(since 1997), some clutches were laid<br />
in deep water, whereas o<strong>the</strong>rs were<br />
deposited in shallow marginal areas,<br />
leading to enhanced variation in water<br />
depth at <strong>the</strong> spawning sites (Figure 5). In<br />
2001, most egg masses were laid close<br />
to <strong>the</strong> water surface.<br />
Spawn <strong>of</strong> R. temporaria was <strong>of</strong>ten<br />
aggregated in clusters, to which new egg<br />
masses were added over several days.<br />
When <strong>the</strong> pools in <strong>the</strong> basin were connected<br />
to running water, spawning <strong>of</strong><br />
this species became restricted to marginal<br />
areas; deep pools were no longer<br />
used, which resulted in an overall shift to<br />
lower mean values for water depth at<br />
<strong>the</strong> spawning site (Figure 5).<br />
In later years, both species ceased to<br />
spawn in water bodies which had<br />
become much deeper (areas A, B). Both<br />
species laid eggs in a bay <strong>of</strong> <strong>the</strong> stream<br />
outside <strong>the</strong> basin (area D) and in shallow<br />
muddy pools in <strong>the</strong> uppermost part <strong>of</strong><br />
<strong>the</strong> basin, which were not connected to<br />
<strong>the</strong> stream (area C; <strong>the</strong>se pools were<br />
mostly dry in 2001).<br />
DISCUSSION<br />
The stream restoration project took some<br />
unexpected turns, such as a catastrophic<br />
flood in summer 1997 during <strong>the</strong> construction<br />
phase, and <strong>the</strong> immigration <strong>of</strong><br />
beavers. Overall, water bodies available<br />
as frog spawning sites in "<strong>the</strong> study area<br />
became much larger, more stable, and<br />
cooler. Numbers and diversity <strong>of</strong> preda-
40 Biota 3/i-a, 2002 GOLLMANN, GOLLMANN, BAUMGARTNER & WARINGER<br />
tors such as fish (including pike, Esox<br />
lucius), birds (mallards, rails, herons) and<br />
aquatic insects have increased greatly.<br />
The effects <strong>of</strong> habitat change on population<br />
dynamics are hard to predict. In <strong>the</strong><br />
short run, connection <strong>of</strong> spawning pools<br />
to <strong>the</strong> stream was certainly advantageous.<br />
O<strong>the</strong>rwise, no tadpoles could<br />
have survived to metamorphosis in <strong>the</strong><br />
very dry years 2000 and 2001. For R.<br />
dalmatina, <strong>the</strong> expansion <strong>of</strong> <strong>the</strong> water<br />
bodies means loss <strong>of</strong> terrestrial habitat;<br />
hence, a reduction in <strong>the</strong> number <strong>of</strong><br />
breeding frogs in <strong>the</strong> study area can be<br />
expected. In <strong>the</strong> long run, decreased<br />
desiccation risk is counteracted by<br />
increased predation risk, but <strong>the</strong> outcome<br />
<strong>of</strong> <strong>the</strong>se opposing trends is uncertain,<br />
and may differ between species. In<br />
some riverine floodplains <strong>of</strong> sou<strong>the</strong>rn<br />
Germany, habitat management that<br />
increased flooded areas or <strong>the</strong> number <strong>of</strong><br />
permanent pools led to growth <strong>of</strong> R.<br />
temporaria populations (Kuhn 2001,<br />
Laufer 2001, Utschick 2001).<br />
Shifting spawning sites to <strong>the</strong> shallow<br />
margins may have two reasons: predator<br />
avoidance or a preference for warm<br />
microhabitats under <strong>the</strong> generally cooler<br />
conditions. Predation on frog eggs<br />
seemed to be negligible in our study area<br />
(unpublished data). Laurila & Aho<br />
(1997) showed that R. temporaria did<br />
not avoid spawning in pools containing<br />
fish which prey on tadpoles. If predation<br />
risk did at all influence spawning site<br />
selection at our study site, adult frogs<br />
may have evaded deep water because <strong>of</strong><br />
<strong>the</strong> presence <strong>of</strong> predatory fish such as<br />
pike.<br />
There was some opportunity for interspecific<br />
competition among brown frog<br />
tadpoles, especially in early study years<br />
with drying pools, but its importance for<br />
larval growth and survival was probably<br />
subordinate to abiotic factors (hydroperiod,<br />
water temperature, water velocity)<br />
and predation.<br />
Acknowledgements<br />
We thank <strong>the</strong> students <strong>of</strong> <strong>the</strong> University<br />
<strong>of</strong> Vienna field course in amphibian ecology<br />
for <strong>the</strong>ir help in collecting and<br />
analysing data. GG acknowledges support<br />
by <strong>the</strong> Austrian Academy <strong>of</strong><br />
Sciences, in <strong>the</strong> framework <strong>of</strong> ecological<br />
monitoring <strong>of</strong> river restructuring commissioned<br />
by <strong>the</strong> Municipality <strong>of</strong> Vienna<br />
(Magistratsabteilung 45, Wasserbau).<br />
REFERENCES<br />
BAUMGARTNER, C., BITSCHI, N., ELLINGER, N., GOLLMANN, B., GOLLMANN, G., KOCK,<br />
M., LEBETH, E. & WARINGER-LOSCHENKOHL, A. 1996: Laichablage und<br />
Embryonalentwicklung von Springfrosch (Rana dalmatina BONAPARTE, 1840)<br />
und Grasfrosch (Rana temporaria LINNAEUS, 1758) in einem syntopen<br />
Vorkommen (Anura: Ranidae). Herpetozoa9: 133-150.<br />
GASC, J.-P., CABELA, A., CRNOBRNJA-ISAILOVIC, J., DOLMEN, D., GROSSENBACHER, K.,<br />
HAFFNER, P., LESCURE, J., MARTENS, H., MARTINEZ RICA, J.P., MAURIN, H.,<br />
OLIVEIRA, M.E. , SOFIANIDOU, T. S. , VEITH, M. & ZUIDERWIJK, A. (eds.)<br />
1997: Atlas <strong>of</strong> Amphibians and Reptiles in Europe. Societas Europaea<br />
Herpetologica and Museum National d'Histoire Naturelle (IEGP/SPN), Paris.<br />
GOLLMANN, G., BAUMGARTNER, C., GOLLMANN, B. & WARINGER-LOSCHENKOHL, A.<br />
1999: Breeding phenology <strong>of</strong> syntopic frog populations, Rana dalmatina and<br />
Rana temporaria, in suburban Vienna. Verhandlungen der Gesellschaft fur<br />
Okologie 29: 357-361.<br />
KUHN, J. 2001: Amphibien in der Wildflusslandschaft der oberen Isar (Bayern):<br />
Auswirkungen der "Teilruckleitung" seit 1990 und des Spitzenhochwassers
GOLLMANN, GOLLMANN, BAUMGARTNER & WARINGER Biota 3/1-2, 2002 41<br />
1999. Zeitschrift fur Feldherpetologie 8: 43-56.<br />
LAUFER, H. 2001: Amphibien in den Poldern Altenheim (Oberrhein, Baden-Wurttemberg):<br />
Bestandsentwicklung und Auswirkung von Hochwassern. Zeitschrift fur<br />
Feldherpetologie 8: 203-214.<br />
LAURILA, A. & AHO, T. 1997: Do female common frogs choose <strong>the</strong>ir breeding habitat to<br />
avoid predation on tadpoles? Oikos 78: 585-591.<br />
RIIS, N. 1988: The present distribution <strong>of</strong> Rana dalmatina and Rana temporaria in sou<strong>the</strong>rn<br />
Scandinavia explained by a <strong>the</strong>ory <strong>of</strong> competitive exclusion. Mem. Soc. Fauna<br />
Flora Fennica 64: 104-106.<br />
ROHRBACH, T. & KUHN, J. 1997: Der Springfrosch (Rana dalmatina) im westlichen<br />
Bodenseeraum 1994-1996: Verbreitung, Bestande, Laichgewasser. In: Krone, A.,<br />
Kiihnel.K.-D. & Berger.H. (eds:). Der Springfrosch (Rana dalmatina)- Okologie<br />
und Bestandssituation. Rana / Sonderheft 2, Rangsdorf: 251-261.<br />
UTSCHICK, H. 2001: Auswirkungen der Staustufe Perach auf die Amphibienbestande der<br />
Aue (Unterer Inn, Bayern) in der Wildflusslandschaft der oberen Isar (Bayern).<br />
Zeitschrift fur Feldherpetologie 8: 119-129.<br />
WILBUR, H.M. 1996: Multistage live cycles. In: Rhodes, O.E., Jr., Chesser, R.K. & Smith,<br />
M.H. (eds.) Population dynamics in ecological space and time. The University <strong>of</strong><br />
Chicago Press. Chicago & London: 75-108.
GROSSENBACHER 3/1-2, 2OO2 43<br />
First results <strong>of</strong> a 20-year-study on<br />
Common Toad Bufo bufo in <strong>the</strong><br />
Swiss Alps<br />
Kurt GROSSENBACHER<br />
Museum <strong>of</strong> Natural History, Bernastrasse 15, CH-3005 Bern<br />
E-mail: groba@nmbe.unibe.ch<br />
Abstract<br />
From 1982 until 2001 a population <strong>of</strong> 150-200 Common Toad Bufo bufo on <strong>the</strong> Grosse<br />
Scheidegg near Grindelwald in <strong>the</strong> Bernese Alps (altitude 1850m) was registered every<br />
spring at <strong>the</strong> breeding site. The toads were marked in <strong>the</strong> first years with toe clipping,<br />
in <strong>the</strong> last 9 years with transponders. The number <strong>of</strong> first time breeders showed a deep<br />
depression in <strong>the</strong> late 80s and recovered in <strong>the</strong> nineteen-nineties. One third <strong>of</strong> all males<br />
and two thirds <strong>of</strong> all females reproduce only once. The maximum reproductive rate <strong>of</strong><br />
a female is seven times within 11 years. Most females reproduced every second year,<br />
but reproduction in two successive years is not rare. The maximum age observed is 25<br />
years, <strong>of</strong> a male marked in 1983 which was present at <strong>the</strong> breeding place during 16<br />
years. The maximum age <strong>of</strong> a female is 20 years. Reproduction success was very low in<br />
<strong>the</strong> nineteen-nineties, but an effect on <strong>the</strong> adult population has not yet appeared.<br />
Keywords: Bufo bufo, reproduction, age, longevity, population dynamics, high altitude,<br />
Swiss Alps.<br />
Received 29 October; accepted 18 December 2001
44 Biota 3/1-2,2002 GROSSENBACHER<br />
INTRODUCTION<br />
Hemelaar (1988) reported a 3-year study<br />
<strong>of</strong> <strong>the</strong> population structure <strong>of</strong> Common<br />
Toad, comparing several populations<br />
throughout Europe. All but one population<br />
were situated in lowlands, <strong>the</strong> only<br />
exception being a population above<br />
Grindelwald in <strong>the</strong> Swiss Alps. The Dutch<br />
herpetologists marked <strong>the</strong> animals with<br />
toe-clipping and analysed age by skeletochronology<br />
(Hemelaar et al. 1987).<br />
Our study picked up where Hemelaar<br />
left <strong>of</strong>f in 1985, focused on <strong>the</strong> same<br />
mountain population, and has continued<br />
through <strong>the</strong> 2001 breeding season. Here<br />
we examine aspects <strong>of</strong> 20 years <strong>of</strong> data<br />
coming from this population.<br />
STUDY AREA<br />
The site <strong>of</strong> this study <strong>of</strong> an alpine<br />
Common Toad population is situated<br />
above Grindelwald in <strong>the</strong> Bernese<br />
Oberland, below <strong>the</strong> pass Grosse<br />
Scheidegg. The altitude <strong>of</strong> <strong>the</strong> pass is<br />
1960 m and <strong>the</strong> pond, at 1850 m, is only<br />
a few meters from <strong>the</strong> road and easily<br />
accessible. The pond measures about 30<br />
x 10 m and <strong>the</strong> maximum depth is<br />
approximately one metre. The vegetation<br />
consists <strong>of</strong> Potamogeton alpinus,<br />
Sparganium angustifolium, Carex rostrata<br />
and Carex fusca and has clearly<br />
increased in cover over <strong>the</strong> last 20 years<br />
(pers. obs.). O<strong>the</strong>r resident amphibian<br />
populations include Triturus alpestris,<br />
Common Toad and a few Rana temporaria,<br />
<strong>the</strong> eggs and larvae <strong>of</strong> which are<br />
usually eliminated by <strong>the</strong> newts.<br />
METHODS<br />
As a basis (1982-1984) we had all <strong>the</strong><br />
individually marked toads <strong>of</strong> Hemelaar<br />
(1988). The initial objective was to follow<br />
<strong>the</strong> life histories <strong>of</strong> <strong>the</strong> marked toads<br />
to <strong>the</strong> end.<br />
We initially avoided individual marking<br />
with toe clips because <strong>of</strong> <strong>the</strong> number <strong>of</strong><br />
toes (4) that needed to be removed.<br />
Instead, we marked newly captured individuals<br />
by removing one toe upon first<br />
date <strong>of</strong> arrival, allowing <strong>the</strong> definition <strong>of</strong><br />
year classes (1985-1992). Later (1993-<br />
2001) we used transponders to identify<br />
individuals.<br />
We visited <strong>the</strong> pond during <strong>the</strong> evening<br />
every 4-5 days, starting each year at <strong>the</strong><br />
beginning <strong>of</strong> snow-melt. It was impossible<br />
to find all toads during daylight<br />
because most were well-hidden in <strong>the</strong><br />
Carex-vegetation. At dusk (9:30 to<br />
10:00 p.m.) we started catching all animals<br />
we observed. The length and <strong>the</strong><br />
weight <strong>of</strong> <strong>the</strong> animals were measured,<br />
<strong>the</strong> toe-code or <strong>the</strong> current transponder<br />
number recorded and first-capture animals<br />
were fitted with a transponder. This<br />
process generally lasted 2-3 hours.<br />
RESULTS<br />
Only several first results can be presented<br />
here; <strong>the</strong> amount <strong>of</strong> results is huge,<br />
and only a small part has been analyzed.<br />
The recapture rate <strong>of</strong> <strong>the</strong> toads from<br />
1985 on, marked individually in 1982-84<br />
(143 males, 121 females), was generally<br />
low in <strong>the</strong> females, never exceeding 9<br />
females per year. The last female <strong>of</strong> this<br />
group was present in 1993 (female no.<br />
411, which came to breed 7 times within<br />
11 years). The decrease <strong>of</strong> <strong>the</strong> males is<br />
slower and more regular, starting with 53<br />
recaptured males in 1985. The longest<br />
living male (No. 418) was regularly present<br />
at <strong>the</strong> breeding place for 16 years<br />
(1983-1998)! In contrast to <strong>the</strong> females,<br />
most <strong>of</strong> <strong>the</strong> long-lived males migrated<br />
every year to <strong>the</strong> site <strong>of</strong> reproduction.<br />
Table 1 presents <strong>the</strong> years <strong>of</strong> breeding <strong>of</strong><br />
those females <strong>of</strong> <strong>the</strong> group 1982-84<br />
who were present at least four times. In<br />
most cases a 2-year-cycle can be recognised.<br />
In some cases <strong>the</strong> females arrived<br />
in 2, and in rare cases 3, consecutive<br />
years. In <strong>the</strong> cases <strong>of</strong> <strong>the</strong> females no.<br />
195, 424 and 428 it is likely that <strong>the</strong>y<br />
were present in at least one year
GROSSENBACHER Biota 3/i-a, ac 45<br />
Table 1. »Life story« <strong>of</strong> individually marked females which were present at least four<br />
times at <strong>the</strong> breeding place. X = present at <strong>the</strong> breeding place, but not weighed; 74:<br />
weight <strong>of</strong> a female before spawning; (55): weight <strong>of</strong> a female after having partially<br />
spawned.<br />
Female<br />
No.<br />
411<br />
151<br />
225<br />
82<br />
195<br />
424<br />
428<br />
1982<br />
X<br />
X<br />
1983<br />
X<br />
X<br />
X<br />
X<br />
1984<br />
X<br />
X<br />
X<br />
1985<br />
72<br />
1986<br />
68<br />
61<br />
50<br />
between 1983 and 1987 or 1988 respectively,<br />
but <strong>the</strong>y escaped our control.<br />
In 1982-84 <strong>the</strong> age <strong>of</strong> 108 males and 66<br />
females was determined by skeletochronology<br />
by Hemelaar et al. (1987).<br />
The combination with our recapture<br />
results enables us to present in Figure 1<br />
<strong>the</strong> final age <strong>the</strong>y reached. Most females<br />
don't get older than 10-12 years. This is<br />
<strong>the</strong> age <strong>of</strong> sexual maturity and <strong>the</strong>y<br />
come only once to breed; few <strong>of</strong> <strong>the</strong>m<br />
get older up to a maximum <strong>of</strong> 20 years.<br />
The males' ages are much more scat-<br />
1987<br />
75<br />
56<br />
48<br />
60<br />
66<br />
1988<br />
76<br />
63<br />
59<br />
70<br />
64<br />
1989<br />
74<br />
66<br />
72<br />
1990<br />
74<br />
64<br />
70<br />
1991<br />
74<br />
73<br />
1992<br />
67<br />
(55)<br />
1993<br />
82 7x<br />
6x<br />
5x<br />
4x<br />
4x<br />
4x<br />
4x<br />
tered; some <strong>of</strong> <strong>the</strong>m reach sexual maturity<br />
at 5-7 years, normally at 8-10 years,<br />
but many get much older, with a maximum<br />
<strong>of</strong> 25 years.<br />
A very high proportion <strong>of</strong> <strong>the</strong> females<br />
(62%) were present only once at <strong>the</strong><br />
breeding place (Figure 2); one quarter<br />
came twice, almost 10% 3 times, a.s.o.<br />
up to 7 times. One third <strong>of</strong> <strong>the</strong> males<br />
were present only once at <strong>the</strong> breeding<br />
place. The number <strong>of</strong> multiple breeders<br />
in males is irregularly scattered over <strong>the</strong><br />
scale up to 16 times. 25 males (17% <strong>of</strong><br />
Figure 1. Final age <strong>of</strong> 108 males and 66 females for which <strong>the</strong> age was determined in<br />
1982-84.<br />
5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25<br />
Age in years
46 Biota 3/i-a, 2002 GROSSENBACHER<br />
Figure 2. Percentage <strong>of</strong> males and females present once, twice, three times a.s.o. at <strong>the</strong><br />
breeding place.<br />
50<br />
40<br />
20<br />
PI n „<br />
5x 7x 8x 9x 10x<br />
all individually followed males) were<br />
present in 8 to16 consecutive years. The<br />
mean value <strong>of</strong> presence for females is<br />
1.5 times, for males 3 times.<br />
DISCUSSION<br />
Most <strong>of</strong> <strong>the</strong> males with determined age<br />
originated from <strong>the</strong> two years 1973 and<br />
1974, and most <strong>of</strong> <strong>the</strong> females with<br />
known age from 1972 (Figure 3).<br />
Hemelaar et al. (1987) considered <strong>the</strong><br />
age structure <strong>of</strong> <strong>the</strong> spawning populations<br />
as unstable. This means a pulsation<br />
<strong>of</strong> <strong>the</strong> age classes occurs along <strong>the</strong> time<br />
axis, depending on <strong>the</strong> time span since<br />
<strong>the</strong> last year with high <strong>of</strong>fspring production.<br />
It seems that in several years before<br />
1972 very few or no <strong>of</strong>fspring survived<br />
(<strong>the</strong> age classes after 1974 were too<br />
young to be sexually mature in 82-84).<br />
When Hemelaar et al. (1987) analysed<br />
<strong>the</strong> population in 1982-84, <strong>the</strong>y found a<br />
relatively young population with no<br />
males older than 13 and no females<br />
older than 18. Several <strong>of</strong> <strong>the</strong>se animals<br />
got much older than <strong>the</strong>se maximum<br />
presence at breeding place<br />
ages in <strong>the</strong> following 10-14 years. An<br />
analysis <strong>of</strong> <strong>the</strong> age structure <strong>of</strong> <strong>the</strong> same<br />
population in <strong>the</strong> early 90s would have<br />
come to a very different result, with a set<br />
<strong>of</strong> old animals, both females and males,<br />
around 20 years old. Thus an age analysis<br />
<strong>of</strong> a toad population always depends<br />
on <strong>the</strong> space <strong>of</strong> time since <strong>the</strong> last year<br />
with a high recruitment. The reproductive<br />
success <strong>of</strong> such populations seems to<br />
vary from one year to <strong>the</strong> next to a very<br />
high extent. In many years <strong>the</strong> production<br />
is nearly zero, as in our case in <strong>the</strong><br />
late 60s until 1971. O<strong>the</strong>rwise, at least a<br />
few old animals should have survived<br />
until 1982-84. We observed a similar<br />
phenomenon during <strong>the</strong> 90s, with no<br />
year evidencing high recruitment. It is<br />
not yet clear if this is a normal fluctuation<br />
or <strong>the</strong> beginning <strong>of</strong> a population decline.<br />
Because <strong>of</strong> <strong>the</strong> long delay between birth<br />
and maturity (males 8-10, females 11-12<br />
years) a decline would only be observable<br />
in adults more than a decade later.<br />
Ano<strong>the</strong>r interesting point is <strong>the</strong> fact that<br />
<strong>the</strong> years <strong>of</strong> high recruitment are differ-
GROSSENBACHER Biota 3/i-a, 2002 47<br />
Figure 3. Year <strong>of</strong> birth for males and females for which <strong>the</strong> age was determined in 1982-<br />
84.<br />
40<br />
35<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
d males<br />
• females<br />
1969 1970 1971 1972 1973 1974 1975 1976 1977 1978 1979 1980<br />
ent in males and females. 1972 (best<br />
year for females) was a cold summer<br />
with temperatures below <strong>the</strong> average<br />
from April to October and a very cold<br />
September. In 1973 (best year for males)<br />
<strong>the</strong> summer was much warmer, with a<br />
very warm August and September. 1974<br />
(good year for males) was again ra<strong>the</strong>r<br />
cold with <strong>the</strong> exception <strong>of</strong> a very warm<br />
August. August is normally <strong>the</strong> time <strong>of</strong><br />
metamorphosis; in cold years it is<br />
September.<br />
According to Piquet (1930), Common<br />
Toad produces a higher proportion <strong>of</strong><br />
females under cold water conditions, in<br />
warmer water a higher proportion <strong>of</strong><br />
males. One possible explanation for our<br />
observations is a temperature-dependent<br />
sex determination. But this can only<br />
be considered as an indication in this<br />
direction. O<strong>the</strong>r explanations, such as a<br />
different survival rate, are possible too.<br />
The result that 2/3 <strong>of</strong> all females breed<br />
only once is not surprising. Kuhn (1994)<br />
observed similar or higher rates <strong>of</strong> mortality.<br />
In <strong>the</strong> lowlands <strong>of</strong> England <strong>the</strong><br />
annual mortality was much higher with<br />
70-85% (Gittins et al. 1985, Reading<br />
1989). But in contrast to <strong>the</strong>se studies,<br />
we never found dead females in <strong>the</strong><br />
pond. We presume that overwintering<br />
stress is responsible for this high mortality.<br />
The cause <strong>of</strong> <strong>the</strong> much higher life<br />
expectancy (20 years) in <strong>the</strong> mountains<br />
than in lowlands (Kuhn 1994: 9 years)<br />
can be found in <strong>the</strong> much shorter activity<br />
time per season which prevails at <strong>the</strong><br />
altitude <strong>of</strong> 1850 m a.s.l., between 3.5<br />
and 5 months. In accordance with this<br />
higher life expectancy it is not surprising<br />
that some females breed more <strong>of</strong>ten (6,<br />
even 7 times) than ever found in <strong>the</strong><br />
lowlands (Kuhn 1994: 5 times). The difference<br />
is ra<strong>the</strong>r small because a 2-yearcycle<br />
is more frequent in <strong>the</strong> mountains<br />
than in <strong>the</strong> plain. A detailed analysis <strong>of</strong><br />
<strong>the</strong> relation between 1-year- and 2-yearcycles<br />
and a possible change <strong>of</strong> it will be<br />
presented in a later paper; more data<br />
must be collected. What is really new<br />
and astonishing about <strong>the</strong>se current<br />
observations is <strong>the</strong> extremely old age<br />
and long presence at <strong>the</strong> breeding place<br />
<strong>of</strong> several males. In most o<strong>the</strong>r studies<br />
males did not reach an age <strong>of</strong> 10 years,<br />
and a presence <strong>of</strong> 8 times or more at <strong>the</strong>
48 3/1-2, 2OO2 GROSSENBACHER<br />
breeding place is consequently impossible.<br />
Because <strong>of</strong> extended longevity <strong>of</strong> toads<br />
in higher altitude populations, an analysis<br />
<strong>of</strong> possible changes in <strong>the</strong> population<br />
structure cannot yet be made. A/lore<br />
research is necessary until <strong>the</strong> age class-<br />
es marked individually by transponders<br />
between 1993 and 95 have reached <strong>the</strong><br />
end <strong>of</strong> <strong>the</strong>ir lifespan. Even after 20 years<br />
<strong>of</strong> population marking, many questions<br />
are still open, and this long-term study<br />
must go on.<br />
Acknowledgements<br />
Thanks to A. Hemelaar for beginning <strong>the</strong> study and sharing data, to Peter Pearman,<br />
who helped me improving this manuscript, and to many colleagues and friends who<br />
helped me in <strong>the</strong> field over this long period, especially Silvia Zumbach and Beatrice<br />
Luscher.<br />
REFERENCES<br />
GITTINS, S.P., KENNEDY, R.I., WILLIAMS, R. 1985: Aspects <strong>of</strong> a population age-structure <strong>of</strong><br />
<strong>the</strong> common toad (Bufo bufo) at Llandrindod Wells Lake, mid-Wales. Brit. J.<br />
Herpetol., London 6: 447-449.<br />
HEMELAAR, A. 1988: Age, growth and o<strong>the</strong>r population characteristics <strong>of</strong> Bufo bufo from<br />
different latitudes and altitudes. Journal <strong>of</strong> Herpetology 22/4: 369-388.<br />
HEMELAAR, A., CLAESSEN, V., WIJNANDS, H. 1987: Enkele karakteristieken van een<br />
voortplantingspopulatie van de gewone pad (Bufo bufo) uit het gebergte van<br />
Zwitserland. Lacerta45:130-139.<br />
KUHN, J. 1994: Lebensgeschichte und Demographie von Erdkrotenweibchen Bufo bufo<br />
bufo (L.). Zeitschrift fur Feldherpetologie 1: 3-87.<br />
PIQUET, J. 1930: Determination du sexe chez les batraciens en fonction de la temperature.<br />
Rev. Suisse Zool. 37:173-281.<br />
READING, CJ. 1989: Annual fluctuations in common toad (Bufo bufo): breeding numbers<br />
at a pond in sou<strong>the</strong>rn England (1980-1889). Abstracts First World Congress <strong>of</strong><br />
Herpetology, 11-19 September 1989, University <strong>of</strong> Canterbury, United Kingdom.
GUICKING, JOCER & WINK Biota 3/1-2,2002 49<br />
Molecular Phylogeography <strong>of</strong> <strong>the</strong><br />
Viperine Snake Natrix maura and<br />
<strong>the</strong> Dice Snake Natrix tessellata:<br />
first results<br />
Daniela GUICKING1, Ulrich JOGER2,<br />
Michael WINK1<br />
1lnstitut fur Pharmazeutische Biologic, Im Neuenheimer Feld 364, 69120 Heidelberg,<br />
Germany<br />
!Hessisches Landesmuseum, Zoologische Abteilung, Friedensplatz 1, 64283 Darmstadt,<br />
Germany<br />
E-mail: daniela.guicking@urz.uni-heidelberg.de<br />
Abstract<br />
First results on phylogeographic patterns, based on complete sequences <strong>of</strong> <strong>the</strong> mitochondrial<br />
cytochrome b gene, are presented for Viperine Snake Natrix maura and Dice<br />
Snake Natrix tessellata. Three major phylogeographic clades are identified in N. maura.<br />
The phylogenetically oldest groups <strong>of</strong> haplotypes are found in Tunisia/Sardinia, and in<br />
Morocco; a third clade comprises all European specimens. Genetic distances (uncorrected<br />
p-distances) are 3.9% to 4.6% between major clades, but do not exceed 1.4%<br />
between different European populations. Due to higher genetic diversity in sou<strong>the</strong>rn<br />
parts <strong>of</strong> <strong>the</strong> European range, it is suggested that <strong>the</strong> Iberian peninsula served as a glacial<br />
refugium, from where France and Northwestern Italy were colonized during postglacial<br />
times.<br />
In N. tessellata, genetic differences between distinct populations reach 8.2%. Major<br />
clades correspond to an apparently relict population in central Greece, to populations in<br />
Egypt/Jordan, Kasakhstan, Eastern Turkey, Crete and most <strong>of</strong> Europe. Within Europe<br />
<strong>the</strong> Balkan region is suggested to have served as a glacial refugium during <strong>the</strong><br />
Pleistocene. Apparently, a few lineages colonized central Europe in postglacial times and<br />
gave rise to <strong>the</strong> populations found today from Romania to Germany. Low but clear<br />
genetic differences between Italian and central European populations suggest that Italy<br />
was colonized by a distinct genetic lineage during <strong>the</strong> late Pleistocene and remained isolated<br />
from central European populations. These first results are consistent with general<br />
hypo<strong>the</strong>ses on phylogeographic patterns and postglacial colonization routes in<br />
European animal and plant species.<br />
Key words: phylogeography, intraspecific diversity, Natrix maura, Natrix tessellata,<br />
cytochrome b<br />
Received 28 September; accepted 26 October 2001
50 Biota 3/i-a, 2002 GUICKING, JOGER & WINK<br />
INTRODUCTION<br />
Intraspecific differentiation <strong>of</strong> a species is<br />
a complex outcome <strong>of</strong> geographic,<br />
demographic and ecological factors that<br />
have operated throughout its evolutionary<br />
history (Walker & Avise 1998). It<br />
should be particularly apparent in taxa<br />
that show only limited mobility. As this is<br />
generally true for reptiles, most studies<br />
<strong>of</strong> intraspecific variability in <strong>the</strong>se vertebrates<br />
provide evidence for <strong>the</strong> existence<br />
<strong>of</strong> distinct lineages or morphotypes that<br />
can be well correlated to geographic<br />
regions (e.g. Emys orbicularis: Fritz 1996,<br />
Lenk et al. 1999, Natrix natrix: Thorpe<br />
1984a, Thorpe 1984b, reviewed in<br />
Kabisch 1999; Elaphe obsoleta: Burbrink<br />
et al. 2000). Designation <strong>of</strong> subspecies is<br />
<strong>the</strong> traditional way to account taxonomically<br />
for high intraspecific diversity and<br />
to distinguish lineages that differ by morphology,<br />
coloration, genetics and geographic<br />
distribution.<br />
The genus Natrix has recently been confined<br />
to <strong>the</strong> water snakes <strong>of</strong> <strong>the</strong> western<br />
palaearctic. It comprises four species:<br />
Viperine Snake Natrix maura, Dice Snake<br />
N. tessellata, Grass Snake N. natrix, and<br />
Big-head European Grass Snake N.<br />
megalocephala. However, <strong>the</strong> species<br />
status <strong>of</strong> N. megalocephala, which is<br />
restricted to a small area east and nor<strong>the</strong>ast<br />
<strong>of</strong> <strong>the</strong> Black Sea (Orlow & Tunijew,<br />
1999), is controversial, due to its similarity<br />
to N. natrix (Bohme 1999). Molecular<br />
phylogenetic data <strong>of</strong> <strong>the</strong> genus Natrix<br />
suggest that N. maura is <strong>the</strong> most ancestral<br />
<strong>of</strong> <strong>the</strong> three main species. N. tessellata<br />
and N. natrix are closely related and<br />
form <strong>the</strong> more derived sister group to N.<br />
maura. A detailed study on this topic will<br />
soon be published by R. Lawson et al.<br />
Of <strong>the</strong> three main species, N. natrix<br />
shows <strong>the</strong> broadest ecological tolerance<br />
and <strong>the</strong> largest distribution area. It<br />
occurs throughout sou<strong>the</strong>rn and central<br />
Europe into England and Scandinavia,<br />
throughout great parts <strong>of</strong> Siberia east-<br />
wards to Lake Baikal, in <strong>the</strong> Middle East<br />
and in <strong>the</strong> most nor<strong>the</strong>rly parts <strong>of</strong> Africa<br />
(Kabisch 1999). N. maura and N. tessellata<br />
are ecologically more restricted and<br />
occupy similar ecological niches. Both<br />
species are highly dependent on aquatic<br />
habitats and need warmer climates than<br />
N. natrix. Accordingly, <strong>the</strong>ir distribution<br />
areas do not extend as far north (see Fig.<br />
2). N. maura is found in northwestern<br />
Africa, on <strong>the</strong> Iberian peninsula, in<br />
sou<strong>the</strong>rn France and in northwestern<br />
Italy (Schatti 1999). The range <strong>of</strong> N. tessellata<br />
extends from Italy throughout <strong>the</strong><br />
Balkan, Middle East, sou<strong>the</strong>rn Asia into<br />
China, and includes nor<strong>the</strong>rn Egypt<br />
(Gruschwitz et al. 1999). Isolated small<br />
populations <strong>of</strong> Dice Snakes are found in<br />
western Germany and in <strong>the</strong> Czech<br />
Republic. The only sympatric populations<br />
<strong>of</strong> Dice Snake and Viperine Snake are<br />
found in northwestern Italy and at Lake<br />
Geneva in Switzerland. However, at Lake<br />
Geneva only N. maura is autochthonous,<br />
whereas N. tessellata was introduced by<br />
man (Mebert 1993).<br />
Intraspecific variability has been studied<br />
to some extent at least in all three<br />
species, but only in N. natrix have different<br />
subspecies been described to<br />
account for <strong>the</strong> high diversity (Thorpe<br />
1984a, Thorpe 1984b, reviewed in<br />
Kabisch 1999). In N. maura no subspecies<br />
are currently recognized<br />
although morphological studies have<br />
found some differences in pholidosis and<br />
colouration between geographically distant<br />
populations. However, <strong>the</strong>se differences<br />
appear too small and too inconsistent<br />
to justify designation <strong>of</strong> subspecies<br />
(Schatti 1999). In N. tessellata two subspecies<br />
are distinguished. The small population<br />
<strong>of</strong> <strong>the</strong> Romanian Black Sea island<br />
<strong>of</strong> Serpilor is separated as N. t heinrothi<br />
(Hecht 1930) from <strong>the</strong> nominate form N.<br />
t. tessellata, which includes all o<strong>the</strong>r<br />
populations <strong>of</strong> <strong>the</strong> Dice snake. According<br />
to Gruschwitz et al. (1999), <strong>the</strong> sub-
GUICKING, JOGER & WINK Biota 3/i-a, 2002 51<br />
species concept <strong>of</strong> N. tessellata needs to<br />
be revised.<br />
We have chosen a phylogeographic<br />
approach based on molecular data to<br />
investigate <strong>the</strong> intraspecific differentiation<br />
<strong>of</strong> Viperine Snake and Dice Snake:<br />
1) to find out whe<strong>the</strong>r intraspecific<br />
genetic differentiation exists, 2) whe<strong>the</strong>r<br />
genetic clades correlate to geographic<br />
regions, and 3) whe<strong>the</strong>r <strong>the</strong> results can<br />
be used to reconstruct a possible<br />
microevolutionary history for <strong>the</strong> species<br />
in Europe. In <strong>the</strong> present contribution we<br />
describe first results based on mitochondria!<br />
cytochrome b DMA sequences.<br />
AAATERIAL AND METHODS<br />
132 Dice Snakes and 80 Viperine Snakes<br />
were sampled covering great parts <strong>of</strong><br />
<strong>the</strong>ir distribution ranges. As a source <strong>of</strong><br />
total genomic DNA, blood samples taken<br />
from <strong>the</strong> caudal vein <strong>of</strong> living snakes, tissue<br />
samples collected from dead animals<br />
or museum material, and shed skins<br />
were used. DNA was isolated from small<br />
aliquots <strong>of</strong> <strong>the</strong> samples following standard<br />
proteinase k and phenol chlor<strong>of</strong>orm<br />
protocols (Sambrook et al. 1989).<br />
Polymerase chain reaction (PCR) was<br />
performed to amplify <strong>the</strong> target<br />
sequence using <strong>the</strong> primers L14724NAT<br />
(5'-GAC CTG CGG TCC GAA AAA CCA-<br />
3') and H16064 (5'-CTT TGG TTT ACA<br />
AGA ACA ATG CTT TA-3'; Burbrink et al.<br />
2000) situated in <strong>the</strong> two RNA-genes<br />
flanking <strong>the</strong> cytochrome b gene in reptiles.<br />
PCR was performed in 50 ul volume<br />
containing 0.75 units <strong>of</strong> Amersham<br />
Pharmacia Biotech Taq polymerase, 0.2<br />
mM <strong>of</strong> each dNTR 50 mM KCI, 1.5 mM<br />
MgCI2, 0.5% Triton x-100, 10 mM Tris-<br />
HCI (pH 8.5) and 0.01% BSA. 10 pmol<br />
<strong>of</strong> each primer and 50-100 ng <strong>of</strong> <strong>the</strong><br />
template DNA were used. After an initial<br />
denaturing step for 4 minutes at 94° C,<br />
31 cycles were performed with denaturing<br />
50 seconds at 94° C, annealing 50<br />
seconds at 52° C, and primer extension<br />
90 seconds at 72° C. A final extension<br />
step <strong>of</strong> 5 minutes at 72° C followed.<br />
PCR products were sequenced directly<br />
with <strong>the</strong> dideoxy chain termination<br />
method (Sanger et al. 1977) using <strong>the</strong><br />
Cycle Sequencing Kit (Amersham<br />
Pharmacia Biotech, RPN 2438/RPN<br />
2538) in combination with fluorescently<br />
labelled primers. For cycle sequencing<br />
primers mt-b2 (5'-GCC CAG AAm GAT<br />
ATT TGT CCT CA, modified from Kocher<br />
et al. 1989), NATf (5'-ACT CAG ATA<br />
TyG ATA AAA TCC C-3'), eNAT (5'-TAG<br />
GCA AAT AGr AAG TAT CAT TCT GG-<br />
3'), and mt-le (5'-TCA AAC CCG AAT<br />
GAT ACT TCC TAT T-3') were used. An<br />
initial denaturing step at 94° C for 3 minutes<br />
was followed by 26 cycles at 94° C<br />
(30 sec) and at 55° C (60 sec).<br />
Fluorescently labelled fragments were<br />
analysed on an automated Sequencer<br />
(Amersham Pharmacia Biotech, ALF-<br />
Express II). The sequences were aligned<br />
manually. Complete cytochrome b gene<br />
sequences (1117 nt) were used for this<br />
study.<br />
Phylogenetic analyses were performed<br />
using <strong>the</strong> program packages MEGA<br />
(Kumar et al. 2001) and PAUP version<br />
4.0b8 (Sw<strong>of</strong>ford 2001). For outgroup<br />
rooting in N. maura <strong>the</strong> cytochrome b<br />
sequence <strong>of</strong> Nerodia fasdata (kindly provided<br />
by R. Lawson) and in N. tessellata<br />
sequences <strong>of</strong> N. maura were used.<br />
Maximum Parsimony analyses were conducted<br />
with <strong>the</strong> heuristic search<br />
approach <strong>of</strong> PAUP using <strong>the</strong> tree-bisection-and<br />
reconnection swapping algorithm.<br />
Bootstrap analyses were performed<br />
with <strong>the</strong> heuristic search<br />
approach. For comparison, Neighbor<br />
Joining and preliminary Maximum<br />
Likelihood trees with estimated parameters<br />
were calculated.
52 Biota 3/i-a, 2002 GUICKING, JOGER & WINK<br />
RESULTS AND DISCUSSION<br />
Natrix maura<br />
Phylogenetic analyses<br />
Neighbor Joining, Maximum Parsimony<br />
and preliminary Maximum Likelihood<br />
analyses yielded similar phylogenetic<br />
trees. The parsimony approach found<br />
eight shortest trees for N. maura phy-<br />
logeny. The strict consensus tree is<br />
shown in Figure 1. The intraspecific comparison<br />
<strong>of</strong> different cytochrome b gene<br />
haplotypes for N. maura yielded 103<br />
variable sites (9.22% <strong>of</strong> all sites), <strong>of</strong><br />
which 82 (7.34%) are parsimony informative.<br />
99% or 100% bootstrap support was<br />
Figure 1. Natrix maura phylogeny: Strict consensus cladogram <strong>of</strong> 8 most parsimonious<br />
trees; lengths 295 steps (Cl = 0.8407, HI = 0.1593, Rl = 0.9226). Nerodia fasciata was<br />
used for outgroup rooting. Bootstrap values (500 replicates) <strong>of</strong> more than 50% are indicated<br />
at bifurcations for major lineages. Numbers following <strong>the</strong> localities indicate more<br />
than one identical sequence from <strong>the</strong> same location.<br />
Natrix<br />
maura<br />
99<br />
100<br />
100<br />
58<br />
97<br />
78<br />
87<br />
r Italy Ponte Organasco<br />
L Italy Voghera 2<br />
Switzerland Lake Geneva 6<br />
Spain Olot 2<br />
Spain Olot 2<br />
Spain Olot 2<br />
Spain Olot<br />
France Cote d'Azur<br />
Switzerland L'Allondon<br />
France Lac Bourget<br />
Italy Varzi<br />
Italy Sta. Maigheriia<br />
Italy Sta. Margherita 2<br />
Spain Amposta<br />
France Camargue<br />
France Niort 3<br />
Spain Pyrenees 2<br />
Spain Galicla 2<br />
France Dpt Herault<br />
France Belcastell<br />
France Lac Salagon 2<br />
Spain Delia de L'Ebre S<br />
p Spain Extremadura<br />
1 *- Spain Extremadura<br />
Spain Extremadura<br />
Spain Extremadura 3<br />
Spain Extremadura<br />
I Spain Extremadura<br />
Portugal Higuera<br />
r- Spain Extremadura<br />
f^- Spain Extremadura<br />
"1— Spain Benissa<br />
'— Spain Extremadura<br />
_r France Lac Salagon 2<br />
Spain Galicia<br />
Spain Extremadura<br />
Spain Benissa<br />
Tunisia Tamerza<br />
Sardinia 2<br />
Tunisia An Nafidah<br />
Tunisia Nabul<br />
Morocco Tabounaht 4<br />
Morocco Haut Atlas<br />
Morocco Haul Atlas 2<br />
Morocco Haut Atlas<br />
Morocco Tabounaht<br />
Morocco Tabounaht 2<br />
Morocco Moyen Atlas<br />
Nerodia fasdaia<br />
"European<br />
Clade"<br />
"Tunisian<br />
Clade"<br />
"Morrocan<br />
Clade"
GUICKING, JOGER & WINK Biota 3/i-a, 2002 53<br />
Figure 2. Location <strong>of</strong> sample sites and major mtDNA clades in Natrix maura (open symbols)<br />
and Natrix tessellata (black symbols). The dashed line indicates <strong>the</strong> distribution<br />
range <strong>of</strong> N. maura in <strong>the</strong> west and N. tessellata in <strong>the</strong> east. Different symbols represent<br />
different genetic clades, in N. maura: square: Moroccan clade, triangle: Tunisian clade,<br />
circle: European clade; in N. tessellata: rhomb: Egyptian clade, star: loannina clade, triangle:<br />
Turkish clade, square: Kazakhstan clade, cross: Crete clade, circle: European<br />
clade. Fat lines suggest approximate boundaries between mtDNA clades.<br />
obtained for three strictly monophyletic<br />
mtDNA clades <strong>of</strong> Viperine Snakes (Figure<br />
1). The three genetic clades correspond<br />
well to geographic locations <strong>of</strong> <strong>the</strong> samples<br />
(Figure 2). One clade is comprised<br />
<strong>of</strong> Viperine Snakes from Tunisia and<br />
Sardinia, one clade is located in<br />
Morocco, and <strong>the</strong> third clade comprises<br />
all European specimens.<br />
The two clades from nor<strong>the</strong>rn Africa are<br />
apparently phylogenetically older than<br />
<strong>the</strong> European clade. However, current<br />
data are not unambiguous in <strong>the</strong> positioning<br />
<strong>of</strong> <strong>the</strong> two African clades.<br />
Depending on <strong>the</strong> algorithm and optimality<br />
criterion used, ei<strong>the</strong>r <strong>the</strong> Tunisian<br />
clade or <strong>the</strong> Moroccan clade appears to<br />
be more ancestral. Bootstrap values <strong>of</strong> a<br />
little less than 50% for ei<strong>the</strong>r <strong>of</strong> <strong>the</strong> two<br />
possibilities fur<strong>the</strong>r indicate that <strong>the</strong> positioning<br />
<strong>of</strong> <strong>the</strong>se clades relative to each<br />
o<strong>the</strong>r cannot be reliably inferred from<br />
<strong>the</strong> cytochrome b data set.<br />
Close genetic relationship between<br />
Sardinian and Tunisian Viperine Snakes<br />
suggest that <strong>the</strong> Viperine Snake was<br />
introduced to Sardinia by man. This has<br />
already been suggested by Schatti<br />
(1999) due to morphological similarities<br />
<strong>of</strong> individuals from <strong>the</strong> two localities.<br />
Haplotypes and pairwise genetic comparisons<br />
Pairwise genetic distances are given as<br />
interclade and intraclade comparisons in<br />
Table 1. The three major clades are separated<br />
from each o<strong>the</strong>r by genetic distances<br />
<strong>of</strong> 3.9-4.6%. Intraclade distances<br />
reach 1.34% between different<br />
European samples. In all instances, <strong>the</strong><br />
interclade differences are greater than<br />
<strong>the</strong> intraclade distances, indicating that<br />
<strong>the</strong> three major clades represent independent<br />
evolutionary lineages.<br />
According to <strong>the</strong> concept <strong>of</strong> <strong>the</strong> molecular<br />
clock, genetic distances can provide<br />
information on <strong>the</strong> divergence time <strong>of</strong><br />
two distinct lineages. With an assumed<br />
evolutionary rate for snake mtDNA <strong>of</strong><br />
1.3% per one million years (de Queiroz
54 Biota 3/i-a, 2002 GUICKING, JOGER & WINK<br />
Table 1. Interclade and maximum intraclade genetic distances (p-distances) in a) Natrix<br />
maura and b) Natrix tessellata, based on complete sequences <strong>of</strong> <strong>the</strong> mitochondrial<br />
cytochrome b gene.<br />
a) Natrix maura<br />
Glade "Tunisian" "Moroccan" "European"<br />
"Tunisian" 0.45%<br />
"Moroccan" 3.9-4.5% 0.90%<br />
"European" 3.9-4.6% 3.9-4.6% 1.34%<br />
b) Nafrix tessellata<br />
Glade "loannina"<br />
"loannina" 0.27%<br />
"Egyptian" 8.0-8.1%<br />
"Kazakhstan" 7.8-8.2%<br />
"Turkish" 7.4-8.1%<br />
"Crete" 7.9-8.1%<br />
"European" 7.3-8.1%<br />
et al. in press, adopted from Macey et al.<br />
1998, who obtained sequence divergence<br />
data for some agamid lizards), we<br />
can cautiously estimate <strong>the</strong> divergence<br />
time <strong>of</strong> <strong>the</strong> three major clades in N.<br />
maura to some 3 to 3.5 million years<br />
ago, i.e., in <strong>the</strong> middle/late Pliocene.<br />
Larger sample sizes within <strong>the</strong> European<br />
clade allow some preliminary statements<br />
on <strong>the</strong> microevolutionary history <strong>of</strong> <strong>the</strong><br />
Viperine Snake in Europe. Intraclade<br />
genetic distances <strong>of</strong> 1.34% or less suggest<br />
that radiation <strong>of</strong> <strong>the</strong> European<br />
ancestor into present haplotypes started<br />
about one million years ago and corresponds<br />
well to <strong>the</strong> major Pleistocene<br />
glaciation cycles. Especially <strong>the</strong> last 700<br />
000 years were dominated by major<br />
glacials with a roughly 100 000 year<br />
cycle that alternated with relatively short<br />
interglacials (reviewed in Hewitt 1996).<br />
The climatic fluctuations <strong>of</strong> <strong>the</strong><br />
Pleistocene are known to have greatly<br />
influenced <strong>the</strong> distribution <strong>of</strong> intraspecific<br />
polymorphism in cold-sensitive species<br />
"Egyptian" "Kazakhstan" "Turkish"<br />
0.27%<br />
7.3-7.7%<br />
7.2-7.8%<br />
7.3%<br />
6.9-7.3%<br />
2.42%<br />
3.7^.9%<br />
6.6-6.8%<br />
6.4-7.0%<br />
3.13%<br />
5.6-6.8%<br />
5.6-7.0%<br />
"Crete" "European"<br />
0%<br />
2.96% 0.80%<br />
<strong>of</strong> European flora and fauna (Hewitt<br />
1996, Taberlet et al. 1998). Pleistocene<br />
climatic conditions caused extinction <strong>of</strong><br />
nor<strong>the</strong>rn populations and contraction <strong>of</strong><br />
range to sou<strong>the</strong>rn refugia during cold<br />
periods as well as northward expansion<br />
from refugia during subsequent warmings<br />
(Hewitt 1996, Taberlet et al. 1998).<br />
Such processes imply successive bottlenecks<br />
and a loss <strong>of</strong> genetic diversity in<br />
nor<strong>the</strong>rn populations, whereas highest<br />
genetic diversity is expected in sou<strong>the</strong>rn<br />
refugia (Taberlet et al. 1998). The main<br />
Pleistocene refugia in Europe were located<br />
on <strong>the</strong> Iberian peninsula, in Italy and<br />
in <strong>the</strong> Balkans (e.g. Emys orbicularis:<br />
Lenk et al. 1999). Postglacial colonization<br />
<strong>of</strong> central Europe originated from<br />
<strong>the</strong> Balkan refugium or from <strong>the</strong> Iberian<br />
peninsula, whereas northward expansion<br />
<strong>of</strong> Italian populations in most species<br />
was prevented by <strong>the</strong> barrier <strong>of</strong> <strong>the</strong> Alps<br />
(Taberlet et al. 1998).<br />
In N. maura, highest genetic diversity -<br />
indicated by distinct haplotypes occur-
GUICKING, JOGER & WINK<br />
ring in <strong>the</strong> same area (e.g. in <strong>the</strong><br />
Extremadura) or identical haplotypes<br />
occurring at geographically distant localities<br />
- is found on <strong>the</strong> Iberian peninsula,<br />
suggesting this as <strong>the</strong> main Pleistocene<br />
refugium. Samples from <strong>the</strong> nor<strong>the</strong>astern<br />
part <strong>of</strong> <strong>the</strong> distribution range, that is<br />
northwestern Italy, Switzerland, <strong>the</strong> Cote<br />
d'Azur and Olot in Spain, on <strong>the</strong> o<strong>the</strong>r<br />
hand, share closely related haplotypes<br />
(Figure 1). These regions were apparently<br />
colonized by a single lineage after <strong>the</strong><br />
last glaciation.<br />
Matrix tessellata<br />
Phylogenetic analyses<br />
In N. tessellata, six major phylogenetic<br />
clades can be distinguished, independent<br />
<strong>of</strong> <strong>the</strong> calculation method. The strict<br />
consensus tree <strong>of</strong> 96 most parsimonous<br />
trees is shown in Figure 3. Within <strong>the</strong><br />
ingroup, 218 variable sites (19.5% <strong>of</strong> all<br />
sites) were found in 1117 nucleotides <strong>of</strong><br />
<strong>the</strong> cytochrome b gene. 183 (16.4%) <strong>of</strong><br />
<strong>the</strong>se are parsimony-informative.<br />
The two most ancestral clades correspond<br />
to samples from Egypt/Jordan and<br />
to an apparently isolated population<br />
from central Greece, Lake loannina<br />
(Figures 2, 3). Most samples from<br />
Turkey, toge<strong>the</strong>r with all samples from<br />
Armenia, Azerbaijan, Georgia and<br />
Kazakhstan, form ano<strong>the</strong>r clade, which is<br />
fur<strong>the</strong>r subdivided and <strong>the</strong>refore is treated<br />
here as two different clades, <strong>the</strong><br />
Kazakhstan clade and <strong>the</strong> Turkish clade.<br />
The best represented group corresponds<br />
to samples from Europe, except for those<br />
from Lake loannina. Because <strong>of</strong> high<br />
genetic distances between European<br />
mainland samples and samples from <strong>the</strong><br />
island <strong>of</strong> Crete (Table 1), we separate <strong>the</strong><br />
Crete samples as a clade <strong>of</strong> <strong>the</strong>ir own.<br />
Approximate geographic boundaries<br />
between different clades are indicated in<br />
Figure 2.<br />
The existence <strong>of</strong> a population with<br />
ancestral haplotypes at Lake loannina is<br />
55<br />
one <strong>of</strong> <strong>the</strong> most surprising results <strong>of</strong> this<br />
study. Several scenarios can be drawn to<br />
explain <strong>the</strong> parapatric occurrence <strong>of</strong> different<br />
haplotypes in Greece. Both lineages<br />
might have descended from a common<br />
ancestor that inhabited Europe several<br />
million years ago and was <strong>the</strong>n biogeographically<br />
separated into different<br />
populations which evolved independently<br />
afterwards. The existence <strong>of</strong> a phylogenetically<br />
ancestral haplotype within<br />
<strong>the</strong> distribution area <strong>of</strong> <strong>the</strong> more derived<br />
European haplotypes might fur<strong>the</strong>r suggest<br />
that Europe was inhabited by a different<br />
genetic lineage <strong>of</strong> Dice Snakes in<br />
<strong>the</strong> far past. This scenario would assume<br />
that <strong>the</strong> early European Dice Snakes<br />
became almost entirely extinct, with <strong>the</strong><br />
only descendants <strong>of</strong> that lineage today<br />
being left at Lake loannina, whereas <strong>the</strong><br />
present European haplotype has<br />
descended from a lineage that colonized<br />
Europe much later. A third possible<br />
explanation is that <strong>the</strong> population at<br />
Lake loannina derives from an anthropogenetically<br />
introduced population.<br />
However, this possibility seems less likely,<br />
because our data do not suggest any<br />
relationship <strong>of</strong> <strong>the</strong> loannina haplotype to<br />
haplotypes found elsewhere, and <strong>the</strong><br />
region <strong>of</strong> Lake loannina is known to<br />
inhabit endemic lineages <strong>of</strong> several o<strong>the</strong>r<br />
organisms as well (P. Kyriakopoulou-<br />
Sklavounou, pers. cornm.).<br />
Haplotypes and pairwise genetic comparisons<br />
Genetic distances between <strong>the</strong> six clades<br />
are 3.0-8.2% (Table 1), suggesting independent<br />
evolution <strong>of</strong> <strong>the</strong> most distant<br />
lineages since <strong>the</strong> early Pliocene.<br />
Intraclade genetic distances are smaller,<br />
reaching a maximum <strong>of</strong> 2.42% and<br />
3.13% in <strong>the</strong> two deeply subdivided<br />
clades <strong>of</strong> Kazakhstan and Turkish samples<br />
(Tab. 1). Samples from Crete are<br />
separated from mainland European samples<br />
by genetic distances <strong>of</strong> up to
56 3/1-2, 2OO2 GUICKING, JOGER & WINK<br />
Figure 3. Matrix tessellata phylogeny: Strict consensus cladogram <strong>of</strong> 96 most parsimonious<br />
trees; lengths 517 steps (Cl = 0.6716, HI = 0.3284, Rl = 0.9197). N. maura<br />
sequences were used for outgroup rooting. Bootstrap values (100 replicates) greater<br />
than 50% are indicated at bifurcations for major lineages. Numbers after <strong>the</strong> location<br />
indicate more than one identical sequence from <strong>the</strong> same location.<br />
71<br />
81<br />
|— 99<br />
100<br />
p Italy Lazio 4<br />
*- Italy Toscana<br />
p Switzerland Lago di Lugano<br />
*- Switzerland Lago di Lugano<br />
85 (~ Hungary Balaton<br />
*— Hungary Balaton<br />
S4 ,f Romania Tulcea<br />
— P- Romania Tulcea 3<br />
*— Romania Tulcea<br />
_ . . _<br />
r Turkey Sarkale 5<br />
J""1- Georgia Batumi<br />
lOpJLr Turkey Malatya<br />
j I *- Armenia Eranus<br />
n\ Azerbaijan Kubatly<br />
92<br />
nn r-T TurKey EsMK>9a<br />
LZZ-P" Turkey Yenioaga 5<br />
|_r Turkey Sinop<br />
^ Turkey Catalzeytin<br />
Turkey Biracik<br />
innrT Kazakhstan Alma-Ata<br />
98 f^P- Kazakhstan Alma-Ata 2<br />
tesseflata 10°<br />
' Georgia Agara<br />
r Eavut<br />
*- Jordan Jarash 2<br />
* «« r Greece Lake loannina<br />
1UU P- Greece Lake loannina<br />
|_r Greece Lake loannina<br />
"- Greece Lake loSnnina<br />
r~~ Nattix maura France Lac Bourget<br />
fj— Natrix maura Italy Ponte Organasco<br />
a<br />
' — • Matrix maura Spain Extremadura<br />
2.96%; <strong>the</strong> divergence time <strong>of</strong> <strong>the</strong> two<br />
lineages could <strong>the</strong>refore be estimated at<br />
a little more than two million years ago,<br />
i.e., at <strong>the</strong> transition <strong>of</strong> Pliocene and<br />
Pleistocene.<br />
Within <strong>the</strong> European Clade, genetic distances<br />
<strong>of</strong> 0.80% suggest that radiation<br />
"European<br />
Clade"<br />
"Crete Clade"<br />
"Turkish<br />
Clade"<br />
"Kazakhstan<br />
Clade"<br />
"Egyptian Clade<br />
"loannina<br />
Clade"<br />
into present haplotypes occurred during<br />
<strong>the</strong> middle and late Pleistocene.<br />
Phylogenetic relationships are, however,<br />
only weakly resolved. Three apparently<br />
independent genetic lineages can be distinguished:<br />
one comprises samples from<br />
Lake Balaton in Hungary, one comprises
GUICKING, JOGER & WINK<br />
samples from Tulcea at <strong>the</strong> mouth <strong>of</strong> <strong>the</strong><br />
Danube river in Romania, and one<br />
includes <strong>the</strong> Italian and Swiss samples. A<br />
fourth lineage that is resolved by<br />
Neighbor Joining and Maximum<br />
Likelihood analyses, but does not appear<br />
in <strong>the</strong> Maximum Parsimony approach,<br />
includes all samples from Germany, <strong>the</strong><br />
Czech Republic and some samples from<br />
Bulgaria and Romania. The Balkan region<br />
most likely served as a Pleistocene<br />
refugium, from where <strong>the</strong> different lineages<br />
postglacially invaded <strong>the</strong> nor<strong>the</strong>rn<br />
parts <strong>of</strong> <strong>the</strong> present distribution area.<br />
A close relationship between Italian and<br />
Balkan specimens indicate that Italy did<br />
not play a major role as a glacial<br />
refugium in <strong>the</strong> Dice Snake, but was<br />
ra<strong>the</strong>r colonized only during <strong>the</strong> late<br />
Pleistocene. Genetic distinction <strong>of</strong> Italian<br />
from central European Dice Snakes<br />
apparently reflects <strong>the</strong> barrier function <strong>of</strong><br />
<strong>the</strong> Alps.<br />
Conclusions<br />
Our data show a remarkable intraspecific<br />
genetic subdivision <strong>of</strong> Viperine Snake<br />
and Dice Snake. Both species are composed<br />
<strong>of</strong> several monophyletic mtDNA<br />
clades that can be well correlated to geographic<br />
regions. Lack <strong>of</strong> co-existence <strong>of</strong><br />
major haplotypes and large genealogical<br />
gaps between most clades suggest long-<br />
Biota 3/i-a, Z, 2OO2 57<br />
term extrinsic barriers to gene flow<br />
(Walker & Avise 1998). Phylogeographic<br />
data from European specimens provide<br />
<strong>the</strong> first information on <strong>the</strong> microevolutionary<br />
history <strong>of</strong> <strong>the</strong> two species in<br />
Europe. To complement <strong>the</strong> existing<br />
data, we would like to include fur<strong>the</strong>r<br />
sample sites, especially from underrepresented<br />
regions and possible suture zones<br />
<strong>of</strong> different lineages. We are also planning<br />
to include nuclear markers in our<br />
study (ISSR-PCR genetic fingerprints, see<br />
Wink et al. 2001) to study possible introgression<br />
between adjacent lineages.<br />
Finally, <strong>the</strong> data presented here provide<br />
evidence for <strong>the</strong> existence <strong>of</strong> distinct<br />
genetic lineages that have evolved independently<br />
over several million years,<br />
both in Viperine Snake and Dice Snake.<br />
Genetic distances between different<br />
clades are comparatively high and lie<br />
within <strong>the</strong> range <strong>of</strong> distances between<br />
closely related species and subspecies in<br />
o<strong>the</strong>r reptiles (Johns & Avise 1998, de<br />
Queiroz et al. in press). Therefore, our<br />
data provide a first basis for a taxonomic<br />
revision <strong>of</strong> <strong>the</strong> two species. Distinction<br />
<strong>of</strong> different lineages, at least at <strong>the</strong> subspecies<br />
level, seems desirable to account<br />
for <strong>the</strong> high intraspecific variability.<br />
However, morphological studies need to<br />
be performed to accomplish such a taxonomic<br />
revision.<br />
Acknowledgements<br />
We are very grateful to all those who have supported and still support our study by collecting<br />
or providing sample material. These are: T. Amann, P. Lenk, H.-P. Eckstein, P.<br />
Godlay, A. Pieh, X. Bonnet, S. Ursenbacher, E. Razzetti, A. Hille, M. Gruschwitz, S. Lenz,<br />
A. Herzberg, W. Fiedler, A. Sproll, A. K. Smole-Wiener, B. Gautschi, D. Modry, M. A. L.<br />
Zuffi, N. Godetsch, G. Mantziou, N. Rastegar-Pouyani, R. Griffiths, D. Ristow, N. Orlov,<br />
S. Tome, A. Kapla, M. Vogrin, U. Utiger and many o<strong>the</strong>rs. Samples from <strong>the</strong> mouth <strong>of</strong><br />
<strong>the</strong> Ebro river in Spain were provided by X. Santos and collected with permission from<br />
<strong>the</strong> "Direccio General del Medi Natural, Generalitat de Catalunya" no. 6539<br />
(26.10.1990). The "Struktur- und Genehmigungsdirektion Nord" in Koblenz, Germany,<br />
gave permission to collect samples <strong>of</strong> German Dice Snakes. We thank R. Lawson for his<br />
constant help with emerging problems and for providing unpublished sequences. A.<br />
Rasmussen and M. Ivanov will probably note that we tried to keep in mind some <strong>of</strong><br />
<strong>the</strong>ir helpful critiques given at <strong>the</strong> <strong>SEH</strong> meeting in Slovenia. Our study is financially sup-
58 Biota 3/1-2, 2002 GUICKING, JOGER & WINK<br />
ported by <strong>the</strong> Deutsche Forschungsgemeinschaft (JO-134/7 and WI-719/18).<br />
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Staatliches Museum fur Tierkunde Dresden 51: 41-49
HAILEY & LAMBERT Biota 3/1-2,2002 61<br />
Comparative growth patterns in<br />
Afrotropical giant tortoises<br />
(genus Geochelone)<br />
Adrian HAILEY1 & Michael R.K. LAMBERT2*<br />
'Department <strong>of</strong> Biological Sciences, University <strong>of</strong> Zimbabwe, PO Box MP167, Mount<br />
Pleasant, Harare, Zimbabwe<br />
Present address: School <strong>of</strong> Biological Sciences, University <strong>of</strong> Bristol, Woodland Road,<br />
Bristol BS8 1UG, United Kingdom<br />
E-mail: adrian.hailey@bristol.ac.uk<br />
"Corresponding author: Natural Resources Institute, University <strong>of</strong> Greenwich at<br />
Medway, Central Avenue, Chatham Maritime, Kent ME4 4TB, United Kingdom<br />
Present address: Environmental Initiatives, Lydbrook House, Upper Lydbrook,<br />
Gloucestershire GL17 9LP, United Kingdom<br />
E-mail: lambertmrk@aol.com<br />
Abstract<br />
Growth to huge sizes is characteristic <strong>of</strong> giant tortoises. Growth data for leopard tortoises<br />
Geochelone pardalis in eastern and sou<strong>the</strong>rn Africa, <strong>the</strong> continental Sahelian<br />
giant tortoise Geochelone sulcata, and <strong>the</strong> insular Aldabran giant tortoise Geochelone<br />
gigantea are reanalysed, and <strong>the</strong>ir growth patterns compared. Geochelone pardalis<br />
shows great body size variation, with size in nor<strong>the</strong>rn populations (Republic <strong>of</strong><br />
Somaliland) comparable to that <strong>of</strong> insular giant tortoises. All species and populations fitted<br />
a two-phase growth curve: an initial asymptotic growth period followed by slow<br />
indeterminate linear growth. Asymptotic growth was non-Bertalanffy in all groups,<br />
most individuals fitting Gompertz or logistic-by-mass models best Juvenile growth (for<br />
ages 3-8) was also approximately linear. Variation in populations and species was in<br />
juvenile growth rate, asymptotic size, rate <strong>of</strong> subsequent indeterminate linear growth,<br />
and old age survival rate. Juvenile growth rates and asymptotic sizes <strong>of</strong> G. pardalis from<br />
Somaliland resembled o<strong>the</strong>r populations, with large mean size due to high survival rates<br />
and continued linear growth after <strong>the</strong> asymptote. Juvenile growth rates and indeterminate<br />
linear growth in Geochelone sulcata resembled those <strong>of</strong> G. pardalis, but larger<br />
asymptotic sizes were reached. Geochelone gigantea had much higher juvenile growth<br />
rates than <strong>the</strong> continental species, and asymptotic size and rate <strong>of</strong> subsequent indeterminate<br />
linear growth were lower in a high-density (Grande Terre) population - with size<br />
achieved by G. sulcata similar - than a low-density (lie Malabar) one. Juvenile growth<br />
rate was unaffected by population density, and relative to larger tortoises was higher in<br />
<strong>the</strong> Grande Terre population, <strong>the</strong> only group to approach Bertalanffy-type growth.<br />
Key words: Africa, asymptotic size, Geochelone, growth curve, growth rate, tortoise<br />
Received 1 September 2001; accepted 23 February 2002
62 Biota 3/1-2,2002 HAILEY & LAMBERT<br />
INTRODUCTION<br />
Growth to huge sizes is perhaps <strong>the</strong> most<br />
striking feature <strong>of</strong> giant tortoises. There<br />
are five Afrotropical species; two are on<br />
<strong>the</strong> mainland continent <strong>of</strong> Africa. The<br />
leopard tortoise Geochelone pardalis has<br />
an extensive north-south range in eastern<br />
and sou<strong>the</strong>rn E. & S. Africa, while <strong>the</strong><br />
African spurred or Sahelian giant tortoise<br />
Geochelone sulcata has an east-west<br />
range across <strong>the</strong> Sahelian region (Iverson<br />
1992).<br />
Geochelone pardalis in populations <strong>of</strong><br />
Somalia's North-West Zone (Republic <strong>of</strong><br />
Somaliland) (hereafter referred to as<br />
Somaliland) have been shown to be substantially<br />
larger than in those fur<strong>the</strong>r<br />
south, especially between <strong>the</strong> Equator<br />
and Tropic <strong>of</strong> Capricorn. Size frequencies<br />
indicate that on average, Somaliland tortoises<br />
even exceed those in <strong>the</strong> eastern<br />
Cape Province, South Africa (Lambert et<br />
al. 1998), where <strong>the</strong> largest individuals<br />
<strong>of</strong> this species have been recorded<br />
(Branch et al. 1990).<br />
Geochelone pardalis in Somaliland experience<br />
wet summers, with copious green<br />
vegetation available as food, and cool<br />
dry winters when refuge is sought and<br />
activity is minimal. A not dissimilar climatic<br />
regime prevails also in <strong>the</strong> Sahelian<br />
region <strong>of</strong> Africa, from Mauritania and<br />
Senegal in <strong>the</strong> west to Eritrea and<br />
Ethiopia in <strong>the</strong> east, coincident with <strong>the</strong><br />
range <strong>of</strong> C. sulcata, <strong>the</strong> world's largest<br />
mainland tortoise. The size <strong>of</strong> this<br />
Sahelian tortoise was approached by<br />
large individual G. pardalis in Somaliland.<br />
Large size could thus be due to <strong>the</strong> similarity<br />
<strong>of</strong> climatic conditions, which may<br />
be particularly favourable for growth in<br />
both <strong>the</strong> Sahelian region and nor<strong>the</strong>rnmost<br />
Somaliland.<br />
The main aim <strong>of</strong> this work, <strong>the</strong>refore,<br />
was to compare growth <strong>of</strong> G. pardalis<br />
tortoises in Somaliland with that <strong>of</strong> G.<br />
pardalis from o<strong>the</strong>r regions in E. & S.<br />
Africa, and also with Sahelian G. sulcata.<br />
In particular, this was to show whe<strong>the</strong>r<br />
large size in Somaliland was due only to<br />
a high growth rate, which could be purely<br />
environmental, or to a different<br />
growth pattern, which is more likely to<br />
reflect genetic differences associated<br />
with speciation.<br />
This alludes to an important question <strong>of</strong><br />
whe<strong>the</strong>r growth differences in tortoises<br />
are phenotypic or genetic. Growth <strong>of</strong><br />
tortoises is not suited to experimental<br />
analysis <strong>of</strong> this problem, because many<br />
differences are based on <strong>the</strong> timing <strong>of</strong><br />
reduced growth at <strong>the</strong> asymptote, and<br />
would require study over several decades<br />
to obtain <strong>the</strong> answer. Any interspecific<br />
differences, such as between G. pardalis<br />
and G. sulcata, must have a genetic<br />
basis. The possible importance <strong>of</strong> phenotypic<br />
effects can be judged by comparison<br />
<strong>of</strong> closely-related populations in different<br />
ecological conditions.<br />
Neighbouring island populations <strong>of</strong> <strong>the</strong><br />
Aldabra giant tortoise Geochelone<br />
gigantea on Aldabra Atoll, Indian Ocean,<br />
differ markedly in population density,<br />
and so growth in this species was examined<br />
in two such populations based on<br />
data in Bourn & Coe (1978). Geochelone<br />
gigantea is one <strong>of</strong> three extant giant tortoise<br />
species inhabiting islands in <strong>the</strong><br />
Indian Ocean, but <strong>the</strong> only one with<br />
substantial populations remaining.<br />
Studies <strong>of</strong> growth in G. pardalis have<br />
been made by several investigators in<br />
Somaliland and elsewhere in E. & S.<br />
Africa (Lambert 1995); Serengeti<br />
National Park, N.E. Tanzania (Lambert et<br />
al. 1998), and in <strong>the</strong> Sengwa Wildlife<br />
Research Area, W. Zimbabwe (Hailey &<br />
Coulson 1999).<br />
An outline <strong>of</strong> main results is given in this<br />
account; data in more detail have been<br />
published elsewhere (Hailey & Lambert<br />
2002).<br />
METHODS<br />
Growth measurements, based on growth
HAILEY & LAMBERT 1-2, 2O02 63<br />
rings, were made on G. pardalis in four<br />
regions <strong>of</strong> eastern and sou<strong>the</strong>rn Africa,<br />
on G. sulcata in <strong>the</strong> western Sahel, and<br />
on C. gigantea on two islets <strong>of</strong> Aldabra<br />
Atoll, Indian Ocean.<br />
The straight-line size measurement (in<br />
mm) used for all three tortoise species,<br />
ei<strong>the</strong>r from direct measurement or calculated<br />
(Bourn & Coe 1978, Lambert 1993,<br />
Lambert 1995, Lambert et al. 1998,<br />
Hailey & Coulson 1999), was <strong>the</strong> midline<br />
straight carapace length (intermarginal<br />
notch to supracaudal scute). Tortoises<br />
were also weighed to within 0.5 kg up to<br />
50 kg, and 1 kg above 50 kg. For consistency,<br />
a single relationship between mass<br />
(in g) and length was used for all populations<br />
<strong>of</strong> C. pardalis.<br />
The growth pattern in chelonians has<br />
been described by a variety <strong>of</strong> ma<strong>the</strong>matical<br />
models. As adopted by Hailey &<br />
Coulson (1999), a number <strong>of</strong> FORD-<br />
WALFORD plots may be used to fit<br />
BERTALANFFY, GOMPERTZ, logistic by<br />
length or logistic by mass growth curves.<br />
This provides a simple method for distinguishing<br />
between major types <strong>of</strong> growth<br />
model, and thus <strong>the</strong> growth pattern in<br />
animals. FORD-WALFORD plots are<br />
especially useful in chelonians, with<br />
growth histories represented by yearly<br />
deposition <strong>of</strong> keratinous rings, and<br />
depend on <strong>the</strong> availability <strong>of</strong> a large<br />
number <strong>of</strong> growth increments to give a<br />
significant negative regression for one or<br />
more growth curves. Excluding <strong>the</strong> possibly<br />
incomplete outer growth ring <strong>of</strong> all<br />
tortoises, only individuals with 10+ increments<br />
(i.e. with 11+ usable growth rings<br />
and 12+ in total) were used.<br />
Following Hailey & Coulson (1999), and<br />
using a common size measure, tortoise<br />
populations were compared by means <strong>of</strong><br />
FORD-WALFORD graphical plots to<br />
determine <strong>the</strong> growth patterns. These<br />
methods were applied to G. sulcata to<br />
show whe<strong>the</strong>r <strong>the</strong> large G. pardalis from<br />
Somaliland had growth more similar to<br />
this species (with which it has a slightly<br />
overlapping geographical range in S. E.<br />
Ethiopia; Lambert 1999) or to o<strong>the</strong>r populations<br />
<strong>of</strong> G. pardalis.<br />
Asymptotic size was estimated from <strong>the</strong><br />
intercept on <strong>the</strong> x axis <strong>of</strong> <strong>the</strong> best-fitting<br />
significant negative regression <strong>of</strong> a<br />
FORD-WALFORD plot on BERTA-<br />
LANFFY, GOMPERTZ or logistic by<br />
length or mass axes.<br />
The BERTALANFFY plot may in some<br />
cases give a significant positive slope<br />
with few data points, where only <strong>the</strong> initial<br />
increasing phase <strong>of</strong> growth is represented.<br />
This is not a fit to <strong>the</strong> BERTA-<br />
LANFFY curve as such, since this requires<br />
decreasing growth rate to an asymptotic<br />
size. A positive slope only would give <strong>the</strong><br />
nonsensical result <strong>of</strong> continuously accelerating<br />
growth to an infinite size.<br />
RESULTS<br />
The Leopard Tortoise Geochelone<br />
pardalis<br />
For G. pardalis, <strong>the</strong> BERTALANFFY<br />
model did not give <strong>the</strong> best fit for any<br />
tortoise in any <strong>of</strong> <strong>the</strong> four regions.<br />
Tortoises from Somaliland and Serengeti<br />
mostly fitted GOMPERTZ (17 individuals)<br />
or logistic by mass (15 individuals)<br />
models best, with <strong>the</strong> remaining seven<br />
individuals fitting <strong>the</strong> logistic by length<br />
model best. Tortoises from E. & S. Africa<br />
mostly fitted <strong>the</strong> GOMPERTZ model best<br />
(15/19 individuals), while most <strong>of</strong> those<br />
from Sengwa fitted <strong>the</strong> logistic by mass<br />
model best (20/23 individuals).<br />
For all regions in which G. pardalis were<br />
measured in sub-Saharan Africa, growth<br />
increments showed an early growth rate<br />
<strong>of</strong> 10-15 mm/yr in years 1-2. Compared<br />
to Somaliland tortoises, <strong>the</strong> growth rate<br />
<strong>of</strong> museum E. & S. Africa tortoises<br />
increased only slightly, to about 15<br />
mm/yr, although <strong>the</strong> variation in growth<br />
rate with age was significant (ANOVA,<br />
F1W56 = 1.89, P < 0.02) over <strong>the</strong> years 1 -<br />
16. The relatively flat growth pattern <strong>of</strong>
64 Biota 3/1-2,2002 HAILEY & LAMBERT<br />
E. & S. Africa tortoises explains why <strong>the</strong><br />
GOMPERTZ model fits <strong>the</strong>se animals<br />
best. In o<strong>the</strong>r regions, after an increasing<br />
rate <strong>of</strong> growth, <strong>the</strong> rate declined in older<br />
animals where <strong>the</strong>re were data available.<br />
For Serengeti and Sengwa tortoises,<br />
asymptotic size in males and females was<br />
significantly different, and so growth<br />
increments were examined separately for<br />
<strong>the</strong> sexes in <strong>the</strong>se two regions.<br />
From growth curves constructed for G.<br />
pardalis from different regions in sub-<br />
Saharan Africa, growth in museum specimens<br />
from E. & S. Africa appeared to be<br />
much slower. However, differences in<br />
growth in <strong>the</strong> o<strong>the</strong>r three regions were<br />
small. After 12 years, <strong>the</strong> divergent<br />
growth data among <strong>the</strong>se regions is due<br />
to small and changing sample sizes.<br />
Asymptotic sizes in G. pardalis from,<br />
respectively, Sengwa, Serengeti, E. & S.<br />
Africa and Somaliland were fairly consistent,<br />
with ascending means <strong>of</strong> 295 ±<br />
S.D. 33, 324 ± S.D. 63, 337 ± S.D. 117<br />
and 344 ± S.D. 113 mm, and <strong>the</strong>re was<br />
no significant difference between <strong>the</strong>m.<br />
Initial sizes at growth ring 0 (hatchling<br />
size), with ascending means <strong>of</strong> 31 ± S.D.<br />
6, 45 ± S.D. 7, 47 ± S.D. 8 and 55 ± S.D.<br />
4 mm in, respectively, E. & S. Africa,<br />
Somaliland, Serengeti and Sengwa tortoises,<br />
did, however, differ significantly<br />
in <strong>the</strong> four regions (F3,?9 = 52.3,<br />
P
HAILEY & LAMBERT Biota 3/i-a, 2002 65<br />
taken up to 27 years for <strong>the</strong> lie Malabar<br />
population and 17 years for <strong>the</strong> population<br />
on Grande Terre. Growth curves for<br />
<strong>the</strong> two C. gigantea populations, one <strong>of</strong><br />
low density (lie Malabar) and <strong>the</strong> o<strong>the</strong>r<br />
<strong>of</strong> high density (Grande Terre), were<br />
compared with that <strong>of</strong> G. sulcata.<br />
Juvenile growth rate <strong>of</strong> G. gigantea over<br />
years 3-6 did not differ between low<br />
density lie Malabar and high density<br />
Grande Terre populations. Juvenile<br />
growth rate at 37.6 mm/yr was almost<br />
twice <strong>the</strong> mean <strong>of</strong> 21.6 ± S.D. 7.9<br />
mm/yr for G. pardalis.<br />
Growth ring increments for G. gigantea<br />
over <strong>the</strong> first 17 years were also used to<br />
fit FORD-WALFORD plots, lie Malabar<br />
mean data fitted <strong>the</strong> GOMPERTZ curve<br />
best, and <strong>the</strong> BERTALANFFY curve least<br />
well; asymptotic size was 613 mm.<br />
Grande Terre data also fitted <strong>the</strong> GOM-<br />
PERTZ curve best, and <strong>the</strong> BERTA-<br />
LANFFY curve least well, but asymptotic<br />
size at 470 mm was much smaller.<br />
Because <strong>of</strong> reduced adult growth, but<br />
normal juvenile growth in <strong>the</strong> dense<br />
population, <strong>the</strong> Grande Terre tortoises<br />
are indeed <strong>the</strong> only ones to which <strong>the</strong><br />
BERTALANFFY curve provides a reasonable<br />
fit, although <strong>the</strong> GOMPERTZ curve<br />
is <strong>the</strong> best fit.<br />
DISCUSSION AND CONCLUSIONS<br />
All species and populations <strong>of</strong> <strong>the</strong><br />
Afrotropical tortoises under discussion<br />
fitted a two-phase growth curve, with an<br />
initial period <strong>of</strong> asymptotic growth up to<br />
about 20 years followed by indeterminate<br />
linear growth. Asymptotic growth<br />
was non-BERTALANFFY in all populations,<br />
and juvenile growth was approximately<br />
linear over years 3-8.<br />
Adult size <strong>of</strong> G. gigantea from <strong>the</strong> dense<br />
Grande Terre population was similar to<br />
that <strong>of</strong> G. sulcata, and to old Somaliland<br />
G. pardalis. Thus all can be grouped as<br />
»giant tortoises*, but <strong>the</strong>y achieve huge<br />
sizes in different ways. The differences lie<br />
in <strong>the</strong> rate <strong>of</strong> juvenile growth, <strong>the</strong><br />
asymptotic size reached, <strong>the</strong> rate <strong>of</strong> linear<br />
growth after this, and <strong>the</strong> duration <strong>of</strong><br />
this period <strong>of</strong> slow growth.<br />
Changes in <strong>the</strong>se variables accounted for<br />
all <strong>the</strong> growth curves observed:-<br />
* Slow juvenile growth - G. pardalis in E.<br />
& N. Africa<br />
* Fast juvenile growth - G. gigantea on<br />
Aldabra.<br />
* Higher asymptote than G. pardalis - G.<br />
sulcata in <strong>the</strong> western Sahel, and G.<br />
gigantea on Aldabra, especially on lie<br />
Malabar<br />
* Faster overall growth than o<strong>the</strong>r tortoises<br />
- G. gigantea on lie Malabar<br />
* Longer-continued late linear growth <strong>of</strong><br />
old animals - G. pardalis in Somaliland.<br />
The high rate <strong>of</strong> growth <strong>of</strong> G. gigantea<br />
may be due to a longer activity season<br />
with tropical maritime conditions on<br />
Aldabra compared to mainland Africa.<br />
Geochelone gigantea on lie Malabar<br />
were active throughout <strong>the</strong> year, while<br />
adults on Grande Terre, due to <strong>the</strong> lack<br />
<strong>of</strong> feeding opportunities in shade, were<br />
inactive for no more than two months<br />
during <strong>the</strong> dry season. In contrast, G.<br />
pardalis at Sengwa and G. sulcata in <strong>the</strong><br />
western Sahel were inactive for five<br />
months <strong>of</strong> <strong>the</strong> dry season.<br />
The reduced body size <strong>of</strong> tortoises in <strong>the</strong><br />
dense Grande Terre population, compared<br />
to those on lie Malabar without<br />
<strong>the</strong> same ecological constraints, is a<br />
proven case <strong>of</strong> environmental effects.<br />
With possible access to a different range<br />
<strong>of</strong> herbaceous food species from adults,<br />
juveniles on Grand Terre appear to be<br />
shielded from competition with adults.<br />
The high growth rates in early years give<br />
a closer approximation to a BERTA-<br />
LANFFY-type growth curve than in any<br />
o<strong>the</strong>r species or population.
66 Biota 3/i-a, 2002 HAILEY & LAMBERT<br />
Acknowledgements<br />
Cost <strong>of</strong> travel and subsistence for M.R.K.L to attend <strong>the</strong> 11th Ordinary General Meeting<br />
<strong>of</strong> Societas Europaea Herpetologica in 2alec (near Celje), Slovenia, 13-17 July 2001,<br />
was covered by <strong>the</strong> Higher Education Funding Council <strong>of</strong> England (HEFCE) through<br />
approval <strong>of</strong> <strong>the</strong> Natural Resources Institute, University <strong>of</strong> Greenwich at Medway<br />
(Chatham), who also provided time for meeting participation, preparation <strong>of</strong> an oral<br />
presentation and manuscript writing for <strong>the</strong> <strong>proceedings</strong> volume.<br />
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HAILEY, A. & LAMBERT, M.R.K. 2002. Comparative growth patterns in Afrotropical giant<br />
tortoises (Reptilia Testudinidae). Tropical Zoology 15: 121-139.<br />
IVERSON, J. B. 1992. A revised checklist with distribution maps <strong>of</strong> <strong>the</strong> turtles <strong>of</strong> <strong>the</strong> world.<br />
Privately printed, Richmond, Indiana, USA.<br />
LAMBERT, M. R. K. 1993. On growth, sexual dimorphism, and <strong>the</strong> general ecology <strong>of</strong> <strong>the</strong><br />
African spurred tortoise, Geochelone sulcata, in Mali. Chelonian Conservation<br />
and Biology 1: 37-46.<br />
LAMBERT, M. R. K. 1995. On geographical size variation, growth and sexual dimor- phism<br />
<strong>of</strong> <strong>the</strong> leopard tortoise, Geochelone pardalis, in Somaliland. Chelonian<br />
Conservation and Biology 1: 269-278.<br />
LAMBERT, M.R.K. 1999. On conservation <strong>of</strong> <strong>the</strong> Sahelian giant tortoise, Geochelone sulcata,<br />
pp. 255-261. In: Miaud, C. & Guyetant, R. (eds.). Current studies in herpetology,<br />
Le Bourget de Lac (<strong>SEH</strong>) [Proceedings <strong>of</strong> <strong>the</strong> 9th Ordinary General<br />
Meeting <strong>of</strong> <strong>the</strong> Societas Europaea Herpetologica, 25-29 August 1998, Le Bourget<br />
du Lac, France]. <strong>SEH</strong>, Le Bourget du Lac, France: 255-261.<br />
LAMBERT, M. R. K., CAMPBELL, K. L. I. & KABIGUMILA ,J. D. 1998. On growth and morphometrics<br />
<strong>of</strong> leopard tortoises, Geochelone pardalis, in Serengeti National Park,<br />
Tanzania, with observations on effects <strong>of</strong> bushfires and latitudinal varia- tion in<br />
populations <strong>of</strong> eastern Africa. Chelonian Conservation and Biology 3: 46-57.
JOVANOVIC, 9URIC & AAARKOVIC BJOta 3/1-2, 2002 67<br />
Tertiary reptiles <strong>of</strong> <strong>the</strong> central<br />
part <strong>of</strong> <strong>the</strong> Balkan peninsula<br />
Miodrag JOVANOVIC, Dragana DURIC &<br />
Zoran AAARKOVIC<br />
Natural History Museum, Njegoseva 51, 11 000 Beograd, Yugoslavia<br />
E-mail: geopal@ptt.yu<br />
Abstract<br />
This paper includes a list, with short remarks, <strong>of</strong> all fossils <strong>of</strong> Tertiary reptiles discovered<br />
in <strong>the</strong> period 1896-2001 in <strong>the</strong> central part <strong>of</strong> <strong>the</strong> Balkan Peninsula, composed <strong>of</strong><br />
Serbia, Republic <strong>of</strong> Srpska and FRY Macedonia. The total number <strong>of</strong> discovered Tertiary<br />
reptile fossils in <strong>the</strong> central part <strong>of</strong> <strong>the</strong> Balkan Peninsula is about 55 taxons in 22 localities.<br />
First recordings are noted for chelonians <strong>of</strong> family Chelidrydae, lizards <strong>of</strong> genus<br />
Ophisaurus and family Gerhonotidae, Mediterranean snakes <strong>of</strong> family Colubridae, large<br />
subtropic snakes <strong>of</strong> genus Vipera, and certain crocodilians.<br />
Key words: Tertiary, chelonians, lizards, snakes, crocodilians, Serbia, Republic <strong>of</strong> Srpska,<br />
Macedonia.<br />
Received 9 January 2002; accepted 15 July 2002
68 3/1-2, 2002 JOVANOVIC, BURIC & MARKOVIC<br />
INTRODUCTION<br />
A first, but incomplete, list <strong>of</strong> Tertiary<br />
reptiles discovered on <strong>the</strong> territory <strong>of</strong> former<br />
SFR Yugoslavia (whose eastern half<br />
we here present under <strong>the</strong> name <strong>of</strong> <strong>the</strong><br />
central part <strong>of</strong> <strong>the</strong> Balkan Peninsula),<br />
was published by Paunovic (1983). From<br />
1983 to <strong>the</strong> present (2001), our knowledge<br />
<strong>of</strong> Tertiary reptiles in this part <strong>of</strong><br />
Europe has become both significantly<br />
different and larger. Within <strong>the</strong> period<br />
1985-2001, Jovanovic, <strong>the</strong> curator <strong>of</strong><br />
<strong>the</strong> Natural History Museum and a biologist,<br />
has surveyed all older accessible<br />
paleoherpetological material from <strong>the</strong><br />
collections <strong>of</strong> <strong>the</strong> Natural History<br />
Museum in Belgrade and o<strong>the</strong>r institutions<br />
in Serbia. These results were published<br />
in several papers (Jovanovic 1989,<br />
1990, 1995a, b). From 1990-1999,<br />
Markovic, a geologist and curator <strong>of</strong> <strong>the</strong><br />
Natural History Museum, discovered by<br />
fieldwork a whole array <strong>of</strong> new localities<br />
with remains <strong>of</strong> Tertiary reptiles. Since<br />
1999, Jovanovic and Markovic have<br />
been joined in paleoherpetological<br />
research by Novakovic, a biologist and<br />
curator <strong>of</strong> <strong>the</strong> Natural History Museum,<br />
who initiated revision <strong>of</strong> all former findings.<br />
This work has resulted in this, <strong>the</strong><br />
most complete list <strong>of</strong> all fossil finds <strong>of</strong><br />
Tertiary reptiles in <strong>the</strong> central part <strong>of</strong> <strong>the</strong><br />
Balkan Peninsula.<br />
METHODS<br />
The authors <strong>of</strong> this paper have revised<br />
older available fossil material and preliminarily<br />
identified new fossil material.<br />
They also provide comments on previous<br />
identifications and reasons behind new<br />
determinations, wherever necessary. In<br />
cases when cited fossil specimens are<br />
known only from literature data, <strong>the</strong><br />
authors did no revision but gave <strong>the</strong>ir<br />
opinion in comments. This is explicitly<br />
stated for all such fossil remains and<br />
those which are known only from credible<br />
oral reports.<br />
Many fossils cited in this paper could not<br />
be determined to species level but only<br />
to genus, family or some higher taxonomy<br />
units. However, regardless <strong>of</strong> this<br />
fact, all <strong>the</strong>se remains were treated as<br />
real species, in both a biological and a<br />
paleontological sense. For <strong>the</strong> specimens<br />
that are presently kept in <strong>the</strong><br />
Paleontological collection <strong>of</strong> <strong>the</strong> natural<br />
history Museum in Belgrade, that was<br />
not explicitly stated since it was understood.<br />
All material was found toge<strong>the</strong>r with<br />
mammal fossils, and <strong>the</strong>refore is dated<br />
according to MN zones (Steininger<br />
1999).<br />
LIST OF LOCALITIES AND RECORDED<br />
SPECIES<br />
EOCENE<br />
Stip (FRY Macedonia):<br />
In this locality, an almost complete shell<br />
<strong>of</strong> a turtle (Temnoclemmys mazedonica<br />
Pasic & Klincarski 1959) was found. This<br />
fossil may be part <strong>of</strong> <strong>the</strong> palentological<br />
collection in <strong>the</strong> Skopje Museum <strong>of</strong><br />
Natural History. This is not only <strong>the</strong> oldest<br />
known record <strong>of</strong> turtles in<br />
Macedonia, but also <strong>the</strong> oldest known<br />
record <strong>of</strong> any Tertiary reptile on <strong>the</strong> territory<br />
<strong>of</strong> <strong>the</strong> central Balkans.<br />
EARLY MIOCENE<br />
Djacki potok. Aleksinac (Serbia) MN1:<br />
Several almost complete turtle shells<br />
were found in vicinity <strong>of</strong> <strong>the</strong> Aleksinac<br />
coal mine (Stevanovic 1969). It is not<br />
known where <strong>the</strong>se fossils are kept<br />
today.<br />
Kraljevo village. Aleksinac (Serbia) MN1:<br />
Three vertebrae, approximately five cm<br />
long (Crocodilia gen. et sp. indet), were<br />
collected by R. Stevanovic.<br />
Dubrava coal mine. Aleksinac (Serbia)<br />
MN1:<br />
According to Stevanovic (1969), at<br />
Djacki Potok and in <strong>the</strong> Dubrava coal
JOVANOVIC, BURl£ & MARKOVIC Biota 3/1-2,2002 69<br />
Table 1. List <strong>of</strong> Tertiary fossils with <strong>the</strong>ir localities and ages in MN zones (for explanation<br />
see text)<br />
fossil<br />
Testudines gen. et sp. indet<br />
Temnoclemmys mazedoniaca<br />
Trionyx sp.<br />
Emys sp.<br />
Mauremmvs serbica<br />
Testudo sp.<br />
Testudo kalksburgensis<br />
Geomyda sp.<br />
Testudinidae gen. et sp. indet<br />
Chelidridae gen. et sp. indet.<br />
Reptilia gen.et sp. indet.<br />
Crocodylia gen.et sp. inded.<br />
Crocodylus mormiensis<br />
Aligarotinae gen.et sp. indet<br />
Tomistoma egsenburgensis<br />
Sauna geaet sp. indet.<br />
Squamata gen.et sp. indet<br />
Vipera sp.<br />
Vijjeridae gen.et sp. indet.<br />
Colubridae gen.et sp. indet<br />
Alethinophidia gen.et sp. indet<br />
Natrix cf. natrix<br />
Natrix sp.<br />
Coluber cf. viridiflaviis<br />
Coluber cfgemonensis<br />
Elaphe cf.aitatorlineata<br />
Elaphe cf. longissima<br />
Elaphe sp.<br />
Chameleontidae gen.et sp. indet<br />
Anguidae gen.et sp. indet<br />
Lacertidae gen.et sp. iadet.<br />
Lacerta sp.<br />
Anguis sp.<br />
Pseudapits sp.<br />
Ophisaums sp.<br />
?Gerrhonotinac gen.et sp. indet<br />
E<br />
1<br />
1<br />
1<br />
MW<br />
2,3<br />
4<br />
i<br />
1<br />
1 3,19<br />
1<br />
1<br />
1<br />
1 5<br />
1<br />
MM4<br />
List <strong>of</strong> localities:<br />
1 - Stip, (FRY Macedonia)<br />
2 - Djacki potok, Aleksinac (Serbia)<br />
3 - Okno Dubrava, Aleksinac (Serbia)<br />
4 - Bogovina (Serbia)<br />
5 - Ugljevik (Republik <strong>of</strong> Srpska)<br />
6 - Jankova Klisura (Serbia)<br />
7 - Sibnica (Serbia)<br />
8 - Milicevo brdo, Krusevica (Serbia)<br />
9 - Jama AAorava (Serbia)<br />
10 - Popovac (Serbia)<br />
11 - Prebreza, Blace (Serbia)<br />
12 - Lazarevac, Trstenik (Serbia)<br />
mine several well preserved turtle<br />
(Testudines) shells were found, with horn<br />
plates and visible markings. Besides <strong>the</strong><br />
shells, specimens also included preserved<br />
limbs. Shell lengths were about 20 cm. It<br />
6<br />
7<br />
7<br />
7<br />
MN5<br />
9<br />
3<br />
9<br />
20<br />
mammal zones<br />
MN<br />
MN6<br />
7+8<br />
21<br />
11,13<br />
13<br />
10<br />
13<br />
10<br />
11<br />
11,12<br />
10,21<br />
10<br />
10 1<br />
10<br />
1<br />
22<br />
14<br />
14<br />
14<br />
14<br />
14<br />
14<br />
MN<br />
mn<br />
15<br />
IS<br />
IS<br />
15<br />
15<br />
MN13 MN14<br />
16<br />
1<br />
17<br />
17<br />
17<br />
17<br />
17<br />
17<br />
17<br />
17<br />
17<br />
17<br />
17<br />
MN16<br />
13 - Lestane (Serbia)<br />
14 - Vracevic, Monastery Bogovadje<br />
(Serbia)<br />
15 - Beluska and Prevalac, Veles<br />
(FRY Macedonia)<br />
16 - Grocka (Serbia)<br />
17 - Kamenjak, Bukulja (Serbia)<br />
18 - Beocin (Serbia)<br />
19 - Village Kraljevo, Aleksinac<br />
(Serbia)<br />
20 - Mala Miliva, Despotovac (Serbia)<br />
21 - Zitni potok, Prokuplje (Serbia)<br />
22 - Village Crvca, Levac (Serbia)<br />
is not known where <strong>the</strong> specimens are<br />
kept today. At <strong>the</strong> same locality limb<br />
bones <strong>of</strong> crocodilians (Crocodylia gen. et<br />
sp. indet) as well as parts <strong>of</strong> joints and<br />
one vertebra were found (Stevanovic P.<br />
IS
70 Biota 3/i-a, 2002 JOVANOVIC, BURIC & MARKOVIC<br />
viva voce, Stevanovic 1969).<br />
Bogovina (Serbia) MN1:<br />
Parts <strong>of</strong> a turtle (Trionyx sp.) shell were<br />
found in <strong>the</strong> Bogovina coal mine<br />
(Laskarev 1949). It is not known where<br />
<strong>the</strong> specimens are kept today.<br />
Ugljevik (Republic <strong>of</strong> Srpska) MN1:<br />
Lignite layers in this Neogene lake in<br />
Dinarides have originated in equivalents<br />
<strong>of</strong> ottnangian (Vujnovic et al 2000). The<br />
bottom <strong>of</strong> Ugljevik lake was gently<br />
descending during <strong>the</strong> accumulation <strong>of</strong><br />
coal, and its water was, at least in one<br />
part <strong>of</strong> <strong>the</strong> period, free-flowing (Krstic<br />
2000). Fauna <strong>of</strong> <strong>the</strong> lake and its shores<br />
was, according to our findings, abundant<br />
in amphibians, birds and mammals that<br />
were subtropical or tropical in character.<br />
Of paleoherpet<strong>of</strong>auna, remains <strong>of</strong> <strong>the</strong><br />
lower jaw <strong>of</strong> a lizard were found. The<br />
teeth resemble those <strong>of</strong> <strong>the</strong> genus<br />
Anguis or Ophisaurus.<br />
Jankova Klisura (Serbia) MN4:<br />
Osteoderms <strong>of</strong> crocodilians (Crocodylia<br />
gen. et sp. indet) were found in a coal<br />
mine (Pavlovic & Djurkovic 1962). It is<br />
not known where <strong>the</strong> specimens are kept<br />
today.<br />
Sibnica (Serbia) MN4:<br />
This locality has been known since 1967<br />
(Petronijevic 1967). Of <strong>the</strong> fossils found,<br />
several were identified: fragments <strong>of</strong><br />
intermaxillar bones and vertebras <strong>of</strong><br />
some lizards (Sauria gen. et sp. indet.), a<br />
fragment <strong>of</strong> <strong>the</strong> lower jaw <strong>of</strong> a<br />
chameleon (Chamaeleontidae gen. et sp.<br />
indet), and a thoracic vertebra <strong>of</strong> a snake<br />
(Vipera sp.).<br />
MIDDLE MIOCENE<br />
Milicevo brdo. Krusevica (Serbia) MN5:<br />
In this locality, part <strong>of</strong> a tortoise (Testudo<br />
sp.) shell was found (Laskarev, 1936). It<br />
is not known where <strong>the</strong> specimen is kept<br />
today. This fragment <strong>of</strong> a Testudo is not<br />
only <strong>the</strong> first find <strong>of</strong> this tortoise in our<br />
region but also <strong>the</strong> first registered find <strong>of</strong><br />
any Tertiary reptile in Serbia.<br />
Jama Morava. Despotovac (Serbia)<br />
MN5:<br />
There are literature data from this locality<br />
(Petronijevic 1967), recorded as part<br />
<strong>of</strong> a turtle shell (Testudines) and a tooth<br />
<strong>of</strong> a small reptile. These fossil remains are<br />
presently kept at <strong>the</strong> Geology faculty. As<br />
<strong>the</strong>re is nei<strong>the</strong>r picture not description,<br />
and <strong>the</strong> place where <strong>the</strong> specimens are<br />
kept is unknown, any kind <strong>of</strong> revision is<br />
impossible.<br />
Mala Miliva. Despotovac (Serbia) MN5:<br />
Fossil osteoderms were found belonging<br />
to certain Anguidae.<br />
Zitni Potok. Macina. Prokuplje (Serbia)<br />
MN6:<br />
In this locality plastron fragments<br />
(Testudines gen. et sp. indet) and a tooth<br />
<strong>of</strong> a crocodilian (Crocodilia) were found<br />
at a depth <strong>of</strong> 11m in <strong>the</strong> sandy-gravel<br />
sediments.<br />
Popovac (Serbia) MN6:<br />
Several fossils <strong>of</strong> various reptiles were<br />
found in a cement marl mine in Popovac.<br />
Among <strong>the</strong>m, turtle remains are <strong>the</strong><br />
most important, and include plastron<br />
fragments <strong>of</strong> Mauremmys serbica<br />
Jovanovic, 1995b and carapace fragments<br />
<strong>of</strong> a tortoise (family Testudinidae).<br />
Overall, <strong>the</strong> most important findings in<br />
reptile pale<strong>of</strong>auna in <strong>the</strong> territory <strong>of</strong> <strong>the</strong><br />
central Balkans are crocodilian remains:<br />
Tomistoma eggenburgensis Toula & Kail<br />
1885, part <strong>of</strong> a jaw with teeth (Pejovic<br />
1951). It is not known where <strong>the</strong> specimen<br />
is kept today. The fossil fragment <strong>of</strong><br />
crocodilian jaw that was identified by<br />
Pejovic through odontological analysis<br />
could not be located by <strong>the</strong> authors <strong>of</strong><br />
this paper, so revision could not be done.<br />
There is a possibility that identification<br />
done by Pejovic is inadequate, as according<br />
to Antunes (1987) and Antunes &<br />
Ginsburg (1989), fossil species <strong>of</strong> genus<br />
Tomistoma can hardly be determined<br />
just by teeth.<br />
Crocodylus moraviensis Jovanovic 1995a<br />
part <strong>of</strong> a skull with jaws. The holotype is<br />
kept at <strong>the</strong> local Museum in Paracin.
I<br />
JOVANOVld, DURIC & MARKOVIC 3/1-2, 2OO2 71<br />
Crocodylidae gen. et sp. indet, several<br />
fragments <strong>of</strong> skull bones, teeth, osteoderms,<br />
etc. (Jovanovic 1989),<br />
Crocodylus sp indet, fragment <strong>of</strong> premaxilla<br />
and nasal bones with preserved<br />
nasal opening. This fossil is presently<br />
kept at <strong>the</strong> Museum in Paracin. As this<br />
fragment shows a very convex rostrum,<br />
it may be assumed that it belonged to<br />
some crocodilian with long jaws, perhaps<br />
a gavial.<br />
Aligatorinae indet, fragments <strong>of</strong> upper<br />
and lower jaw with teeth (Jovanovic,<br />
1995a). The fossil is presently kept in <strong>the</strong><br />
Museum in Paracin. The size <strong>of</strong> <strong>the</strong> angle<br />
between <strong>the</strong> left and right branches <strong>of</strong><br />
<strong>the</strong> jaws enables us to conclude that this<br />
fossil remain belonged to a crocodilian<br />
with wide and short jaws, such as an alligator.<br />
Prebreza. vicinity <strong>of</strong> Blace (Serbia) MN6:<br />
Vegetation <strong>of</strong> <strong>the</strong> locality where reptiles<br />
lived was <strong>of</strong> savannah type. Precipitation<br />
was likely to be rare and sporadic. Also<br />
sporadic were savannah bushfires that<br />
drove animals into lakes where <strong>the</strong>y<br />
drowned and later fossilized (Milosevic<br />
1967).<br />
Turtle remains from genus Trionyx sp<br />
were identified. These were carapax<br />
fragments belonging to one small and<br />
one large turtle. They differ from each<br />
o<strong>the</strong>r not only in size but also by morphology<br />
<strong>of</strong> costalia (bone shape and<br />
sculptured look <strong>of</strong> <strong>the</strong>ir upper surfaces).<br />
Besides, <strong>the</strong>se two bones were found in<br />
two differently coloured and differently<br />
granulated sandstone fragments. This<br />
points ei<strong>the</strong>r to possible paleoecological<br />
differences in habitats <strong>of</strong> <strong>the</strong>se turtles (if<br />
<strong>the</strong> places where both specimens were<br />
found were primary ones) or to <strong>the</strong> existence<br />
<strong>of</strong> some greater time distance<br />
between <strong>the</strong>m. Because <strong>of</strong> morphological<br />
and most probably paleoecological<br />
differences, we have decided to treat<br />
<strong>the</strong>se remains as two different species.<br />
Also in this locality, remains were found<br />
<strong>of</strong> two plastrons <strong>of</strong> certain larger, most<br />
probably aquatic turtles that are presently<br />
kept at <strong>the</strong> paleontological collection<br />
<strong>of</strong> <strong>the</strong> Natural History Museum in<br />
Belgrade. The size, shape and morphology<br />
<strong>of</strong> <strong>the</strong>se plastrons are similar to those<br />
<strong>of</strong> turtles in <strong>the</strong> Chelidridae family.<br />
Turtle remains (plastron fragments, hand<br />
and fossilized eggs) described in <strong>the</strong><br />
paper by Milosevic (1967) were determined<br />
to <strong>the</strong> order level (Testudines);<br />
however, <strong>the</strong>re is some doubt regarding<br />
Milosevic's taxonomic determination.<br />
The plastron fragments were lost and<br />
revision is impossible, while morphoanatomic<br />
analysis <strong>of</strong> <strong>the</strong> fossil hand<br />
remains (number <strong>of</strong> phalangae in <strong>the</strong> fingers<br />
and size) show that it probably does<br />
not belong to a chelonian. The »turtle<br />
eggs« as <strong>the</strong>y are called by Milosevic<br />
(1967) are actually only <strong>the</strong>ir moulds,<br />
almost without any traces <strong>of</strong> shell.<br />
During <strong>the</strong> revision <strong>of</strong> all fossil material<br />
brought earlier from Blace and additional<br />
sieving <strong>of</strong> sediment, bird remains were<br />
discovered. Egg size and distribution<br />
within <strong>the</strong> sediment have repeatedly<br />
reminded <strong>the</strong> authors <strong>of</strong> bird eggs within<br />
a nest. As bird remains were found in<br />
<strong>the</strong> same layer at <strong>the</strong> same locality, it is<br />
probable that <strong>the</strong> »turtle eggs« from<br />
Blace are actually a nest <strong>of</strong> eggs <strong>of</strong> some<br />
Miocene water bird. The <strong>the</strong>ory that fossil<br />
eggs from Blace belong to a turtle was<br />
also doubted by Mikhailov id his paper<br />
on classification <strong>of</strong> fossil amniote eggs<br />
(Mikhailov 1991).<br />
Lazarevac. Trstenik (Serbia) MN6:<br />
Horn plates remains <strong>of</strong> a reptile and few<br />
osteoderms <strong>of</strong> undetermined Anguidae<br />
were found in <strong>the</strong> village <strong>of</strong> Lazarevac in<br />
2001. They are now kept in <strong>the</strong> Natural<br />
History Museum in Belgrade.<br />
Lestane (Serbia) MN6:<br />
In this locality fragments <strong>of</strong> three chelonian<br />
genera were found (Trionyx sp.,<br />
Testudo sp., Emys sp.). The fossil<br />
remains <strong>of</strong> turtles were found in <strong>the</strong> layer
72 Biota 3/i-a, 2002 JOVANOVIC, BURIC & MARKOVIC<br />
<strong>of</strong> Lower Sarmate, but <strong>the</strong>re is a possibility<br />
that <strong>the</strong>y were moved <strong>the</strong>re from<br />
some older, for example Badenian, sediments<br />
(Jovanovic 1998).<br />
Vracevici near Bogovadje Monastery<br />
(Serbia) MN7+8:<br />
This locality is situated at <strong>the</strong> place<br />
reached by Paratethys during <strong>the</strong> mid<br />
Miocene (Prisjazhnjuk et al 2000). The<br />
locality was ei<strong>the</strong>r a small lake or a<br />
waterhole. According to <strong>the</strong> mammal<br />
fossils also discovered here, it may be<br />
concluded that <strong>the</strong> climate was moist<br />
and warm, tropical or subtropical.<br />
Several jaw fragments from <strong>the</strong><br />
Lacertidae family were found, as well as<br />
osteoderms and skull parts <strong>of</strong> <strong>the</strong><br />
Anguinae subfamily or <strong>the</strong><br />
Gerrhonotinae subfamily and <strong>the</strong> vertebrae<br />
<strong>of</strong> snakes (Colubridae and<br />
Viperidae).<br />
Village Crnca. Levac (Serbia) MN7+8:<br />
Fragments <strong>of</strong> upper and lower jaws <strong>of</strong><br />
unknown reptiles (Sauria gen. et sp.<br />
indet.) were collected in 1975 by <strong>the</strong><br />
geologist Dolic.<br />
LATE MIOCENE<br />
Beluska and Prevalec. vicinity <strong>of</strong> Veles<br />
(Macedonia) MN11-J-12:<br />
The area where <strong>the</strong> reptiles found here<br />
lived was similar to <strong>the</strong> present-day<br />
African savannah. Around <strong>the</strong> lake in <strong>the</strong><br />
valley <strong>the</strong>re was a lot <strong>of</strong> grass and bushes.<br />
Vegetation was present mostly<br />
around water basins, lakes and rivers<br />
(Pavlovic & Markovic 1990).<br />
Among <strong>the</strong> fossils found were tortoise<br />
(Testudo sp, unknown where <strong>the</strong> specimen<br />
is kept), lizard Lacerta sp. (Ciiric<br />
1957 also not known where it is kept), as<br />
well as <strong>the</strong> right mandible, left maxilla<br />
and osteoderms <strong>of</strong> Pseudopus sp. Fossil<br />
remains <strong>of</strong> this lizard from Veles were<br />
described for <strong>the</strong> first time in a paper by<br />
Pavlovic & Markovic (1990). In that<br />
paper, <strong>of</strong> all <strong>the</strong> material found, only <strong>the</strong><br />
osteoderms were determined to be <strong>the</strong><br />
remains <strong>of</strong> a lizard Ophisaurus panonni-<br />
cus, or as accepted here, Pseudopus<br />
panonnicus.<br />
The right mandible <strong>of</strong> a lizard from Veles<br />
seems to have been discovered by<br />
Pavlovic and Laskarev after WWI; however,<br />
<strong>the</strong>y nei<strong>the</strong>r identified it nor mentioned<br />
it in reports. During later repeated<br />
sieving <strong>of</strong> sediments, Markovic &<br />
Pavlovic discovered <strong>the</strong> left maxilla and<br />
several osteoderms. From an odontological<br />
viewpoint, <strong>the</strong> mandible and maxilla<br />
<strong>of</strong> <strong>the</strong> fossil Pseudopus sp from Veles are<br />
very similar to each o<strong>the</strong>r and probably<br />
belonged to two individuals <strong>of</strong> <strong>the</strong> same<br />
species. On <strong>the</strong> o<strong>the</strong>r hand, <strong>the</strong> jaws and<br />
teeth <strong>of</strong> Pseudopus sp from Veles differ<br />
morphologically from those <strong>of</strong> Middle<br />
European fossil Pseudopus pannonicus<br />
as well as from <strong>the</strong> recent Mediterranean<br />
Pseudopus apodus.<br />
At <strong>the</strong> same locality, fragments <strong>of</strong> one<br />
larger and one smaller snake vertebra<br />
were found. The osteological characters<br />
point out that <strong>the</strong>se are vertebra <strong>of</strong> a<br />
viper, most probably two different individuals,<br />
and trunk vertebra <strong>of</strong> an<br />
Alethinophidia. Based on collected material<br />
and reports, during <strong>the</strong> excavations<br />
in Veles, Pavlovic and Laskarev paid most<br />
attention to larger fossil remains while<br />
neglecting smaller ones. Therefore, <strong>the</strong>ir<br />
finding and mentioning <strong>of</strong> reptile vertebra<br />
(Reptilia gen. et sp. indet, PavlovL<br />
1922) shows that it was large, that is,<br />
belonging to a very large specimen <strong>of</strong><br />
lizard or snake. Also, two vertebra fragments<br />
were found that were unusual in<br />
shape (elongated and large, with broken<br />
ventral parts) and determined to be<br />
Reptilia gen.et sp. indet. As <strong>the</strong>re was no<br />
o<strong>the</strong>r material to compare <strong>the</strong>m with,<br />
<strong>the</strong>y could not be defined more precise-<br />
ly'<br />
Grocka (Serbia) MN13: *<br />
Part <strong>of</strong> an epiplastron was found<br />
(Geoemyda sp.) This fragment is insufficient<br />
to precisely define <strong>the</strong> turtle<br />
species.
JOVANOVIC, BURld & MARKOVIC Biota 3/1-2,2002 73<br />
EARLY PLIOCENE<br />
Kamenjak (Serbia) MM 14:<br />
This locality is very rich in various fossil<br />
remains <strong>of</strong> herpet<strong>of</strong>auna. The following<br />
items were identified: fragments <strong>of</strong> parietal<br />
bones, jaws and vertebrae <strong>of</strong> Lacerta<br />
sp., many vertebra <strong>of</strong> Anguis sp, osteoderms<br />
<strong>of</strong> Ophisaurus sp, fragments <strong>of</strong><br />
right maxilla <strong>of</strong> Anguis or Ophisaurus as<br />
well as many torsal vertebra <strong>of</strong> snakes<br />
(Natrix sp., Coluber sp., Elaphe sp.,<br />
Vipera sp.).<br />
MIDDLE PLIOCENE<br />
Beocin (Serbia) MN16:<br />
In this locality, part <strong>of</strong> a turtle shell were<br />
found and identified as Testudo kalksburgensisTou\a,<br />
1896, (Mlynarski 1966).<br />
The fossil remains <strong>of</strong> this tortoise are<br />
kept at <strong>the</strong> Geological Institute in<br />
Hungary. Testudo kalksburgensis from<br />
Beocin is <strong>the</strong> first fossil find <strong>of</strong> a turtle<br />
registered in Vojvodina.<br />
DISCUSSION AND CONCLUSION<br />
At <strong>the</strong> end <strong>of</strong> this paper, we must mention<br />
that some <strong>of</strong> <strong>the</strong> fossil remains<br />
determined here to <strong>the</strong> genus level will<br />
be fur<strong>the</strong>r described in later papers and<br />
determined to <strong>the</strong> species level. Some <strong>of</strong><br />
<strong>the</strong>m represent new species for science.<br />
In some <strong>of</strong> <strong>the</strong> earlier papers it is mentioned<br />
that almost all fossil remains <strong>of</strong><br />
reptiles in this part <strong>of</strong> Europe were found<br />
accidentally, for example, while excavating<br />
mammal fauna, and that <strong>the</strong>y were<br />
never considered very important.<br />
However, reptiles and amphibians are<br />
very good ecological indicators, that is,<br />
<strong>the</strong>y are susceptible to various climatic<br />
changes (especially temperature and<br />
humidity). Better knowledge <strong>of</strong> fossil<br />
reptiles in <strong>the</strong> central Balkans would<br />
surely enable us to solve a whole array <strong>of</strong><br />
paleoecological and paleogeographic<br />
problems.<br />
In <strong>the</strong> paper "Miocene Crocodilians <strong>of</strong><br />
Serbia," read at <strong>the</strong> third symposium on<br />
fauna <strong>of</strong> Serbia (Jovanovic 1989), <strong>the</strong><br />
author shared <strong>the</strong> hypo<strong>the</strong>sis that<br />
remains <strong>of</strong> Tertiary reptiles should be<br />
looked for at <strong>the</strong> same places where<br />
remains <strong>of</strong> Tertiary mammals were<br />
found, especially on <strong>the</strong> south exposures<br />
<strong>of</strong> <strong>the</strong> localities. During <strong>the</strong> last several<br />
years, Markovic has found remains <strong>of</strong><br />
small mammals toge<strong>the</strong>r with fossils <strong>of</strong><br />
chelonians, lizards and snakes at almost<br />
all visited Tertiary and Quaternary localities<br />
in Serbia. This preliminary research<br />
has already partially confirmed <strong>the</strong><br />
above-mentioned assumptions <strong>of</strong> <strong>the</strong><br />
authors.<br />
This list includes various species <strong>of</strong><br />
Tertiary reptiles found in various Tertiary<br />
localities in <strong>the</strong> central part <strong>of</strong> <strong>the</strong><br />
Balkans. However, <strong>the</strong> realistic number<br />
<strong>of</strong> species <strong>of</strong> Tertiary reptiles that inhabited<br />
this part <strong>of</strong> Europe was certainly tens<br />
<strong>of</strong> times larger than those we have mentioned<br />
(Jovanovic 1996 a, b, 1998).<br />
Unfortunately, here we must repeat and<br />
stress that only future, systematic paleoherpetological<br />
research in <strong>the</strong> central<br />
Balkans can give a realistic picture <strong>of</strong> <strong>the</strong><br />
Tertiary reptiles <strong>of</strong> this region.
74 Biota 3/1-2, 2002 JOVANOVld, 0UR|£ & MARKOVlC<br />
Acknowledgements<br />
The authors <strong>of</strong> this paper are especially grateful to our colleagues, mineralogist Voislav<br />
Simic and geologist Gordana Jovanovic, for shared data, suggestions, and advice.<br />
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Biota 3/1-2,2002 77<br />
Herpet<strong>of</strong>auna <strong>of</strong> Round Island,<br />
Mauritius<br />
Zoltan KORSOS1 & Balazs TROCSANYI2<br />
'Department <strong>of</strong> Zoology, Hungarian Natural History Museum, Baross u. 13, H-1088<br />
Budapest, Hungary<br />
E-mail: korsos@zoo.zoo.nhmus.hu<br />
department <strong>of</strong> Conservation, Danube-Drava National Park Directorate, Tettye ter 9,<br />
H-7625 Pecs, Hungary<br />
E-mail:balazs.trocsanyi@ktm.x400gw.itb.hu<br />
Abstract<br />
Round Island, north <strong>of</strong> Mauritius in <strong>the</strong> Indian Ocean, is inhabited by eight species <strong>of</strong><br />
reptiles (no amphibians), five <strong>of</strong> which are endemic. The original vegetation <strong>of</strong> <strong>the</strong><br />
island was clear-cut in <strong>the</strong> 18th century. Introduced goats and rabbits were eliminated<br />
relatively recently to protect <strong>the</strong> remnants <strong>of</strong> <strong>the</strong> palm savannah, <strong>the</strong>n <strong>the</strong> whole island<br />
was designated as a nature reserve. A Hungarian team visited <strong>the</strong> island in November<br />
1999 and April 2001, to carry out herpet<strong>of</strong>aunal assessments related to all eight species.<br />
Whereas no specimen <strong>of</strong> <strong>the</strong> Burrowing Boa was encountered, good populations <strong>of</strong><br />
Telfair's and Bojer's Skinks, as well as <strong>of</strong> <strong>the</strong> Ornate Day Gecko, could be estimated.<br />
Several specimens <strong>of</strong> DurrelPs Night Gecko, <strong>the</strong> Keel-scaled Tree Boa, and 33 specimens<br />
<strong>of</strong> Gun<strong>the</strong>r's Gecko were observed.<br />
Key words: Round Island, Mauritius, herpet<strong>of</strong>auna, endemic reptiles, conservation<br />
Received 26 February 2002; accepted 27 July 2002
78 Biota 3/i-a, 2002 KORSOS & TROCSANYI<br />
INTRODUCTION<br />
The tiny, volcanic Round Island with its<br />
151 hectares lies 20 km to <strong>the</strong> north <strong>of</strong><br />
Mauritius in <strong>the</strong> Indian Ocean. It is<br />
inhabited by eight species <strong>of</strong> reptiles (no<br />
amphibians), five <strong>of</strong> which are endemic,<br />
occurring nowhere else in <strong>the</strong> world.<br />
The original vegetation, hardwood<br />
ebony and teak forests, was clear-cut in<br />
<strong>the</strong> 18th century. Introduced goats and<br />
rabbits were eliminated relatively recently<br />
(in 1979 and 1986, respectively) to<br />
protect <strong>the</strong> remnants <strong>of</strong> <strong>the</strong> palm savannah,<br />
<strong>the</strong>n <strong>the</strong> whole island was designated<br />
as a nature reserve with extremely<br />
limited access for scientific research only.<br />
In 1989, a conservation management<br />
plan was set up for <strong>the</strong> island to restore<br />
its vegetation, to control <strong>the</strong> invasion <strong>of</strong><br />
alien species, and to monitor <strong>the</strong> population<br />
changes <strong>of</strong> its native and endemic<br />
flora and fauna.<br />
The aim <strong>of</strong> this paper is to summarize in<br />
brief <strong>the</strong> information presently available<br />
on <strong>the</strong> remarkable herpet<strong>of</strong>auna <strong>of</strong><br />
Round Island, to give an illustrative list <strong>of</strong><br />
<strong>the</strong> eight known reptile species, and to<br />
provide a comprehensive literature<br />
(without references) to initiate fur<strong>the</strong>r<br />
herpetological studies. The scientific<br />
results <strong>of</strong> our expeditions will be published<br />
elsewhere.<br />
MATERIAL AND METHODS<br />
With <strong>the</strong> help <strong>of</strong> <strong>the</strong> local authorities, <strong>the</strong><br />
National Parks and Conservation Service<br />
(NPCS) <strong>of</strong> <strong>the</strong> Ministry <strong>of</strong> Agriculture,<br />
Food, Technology & Natural Resources,<br />
and <strong>the</strong> Mauritian Wildlife Fund (MWF),<br />
a Hungarian team visited <strong>the</strong> island in<br />
November 1999, and later, with <strong>the</strong> support<br />
<strong>of</strong> Fauna and Flora International, in<br />
April 2001. Five and ten days were spent<br />
on Round Island, respectively, <strong>the</strong> necessary<br />
material being carried over by governmental<br />
helicopters, following <strong>the</strong><br />
strict expedition protocol set up previously<br />
by NPCS and MWF. There is no<br />
permanent research station on <strong>the</strong> island<br />
at present, so expedition members have<br />
to bring everything <strong>the</strong>mselves (incl.<br />
food, fresh water, tents, sleeping bags,<br />
etc.), and, similarly, have to remove all<br />
waste material when leaving <strong>the</strong> island.<br />
Usually <strong>the</strong>re are four management<br />
expeditions organized by NPCS every<br />
year, with <strong>the</strong> main purpose being to<br />
monitor plant reintroduction and vegetation<br />
recovery.<br />
Surveys and censuses <strong>of</strong> herpet<strong>of</strong>auna<br />
were made every <strong>full</strong> day during <strong>the</strong><br />
early morning, late afternoon and<br />
evening hours; midday was left out<br />
because <strong>of</strong> <strong>the</strong> extremely hot period<br />
when all reptiles appeared to rest.<br />
Gun<strong>the</strong>r's geckos were caught and<br />
measured on all occasions if possible; <strong>the</strong><br />
same applied for Casarea dussumieri,<br />
whereas o<strong>the</strong>r geckos and skinks<br />
observed were only noted during <strong>the</strong><br />
census. In <strong>the</strong> second visit to <strong>the</strong> island,<br />
special methods were used to rediscover<br />
Bolyeria multocarinata; <strong>the</strong> results will be<br />
discussed in ano<strong>the</strong>r paper.<br />
RESULTS AND DISCUSSION<br />
Round Island reptiles include three geckos<br />
(Gekkonidae), three skinks<br />
(Scincidae), and two snake species (<strong>the</strong><br />
only known members) <strong>of</strong> <strong>the</strong> family<br />
Bolyeriidae, according to <strong>the</strong> following<br />
list (endemic forms are marked by an<br />
asterisk):<br />
Gekkonidae:<br />
Gun<strong>the</strong>r's Gecko Phelsuma guen<strong>the</strong>ri<br />
Boulenger, 1885*<br />
Ornate Day Gecko Phelsuma ornata<br />
Gray, 1825<br />
Durrell's Night Gecko Nactus serpensinsula<br />
durrelli Arnold & Jones, 1994*<br />
Scincidae:<br />
Telfair's Skink Leiolopisma telfairii<br />
(Desjardins, 1831)*<br />
Bojer's Skink Congylomorphus bojerii<br />
(Desjardins, 1831)<br />
Bouton's Skink Cryptoblepharus boutonii
KORSOS & TR6CSANYI BJOta Ti-a. 2002 79<br />
(Desjardins, 1831)<br />
Bolyeriidae:<br />
Keel-scaled Tree Boa Casarea dussumieri<br />
(Schlegel, 1837)*<br />
Burrowing Boa Bolyeria multocarinata<br />
(Boie, 1827)*<br />
Whereas no specimen <strong>of</strong> <strong>the</strong> Burrowing<br />
Boa was encountered (<strong>the</strong> last observation<br />
<strong>of</strong> this species dates back to 1974)<br />
during our two expeditions, relatively<br />
good populations <strong>of</strong> Telfair's and Bojer's<br />
Skinks, as well as <strong>of</strong> <strong>the</strong> Ornate Day<br />
Gecko could be estimated. Several specimens<br />
<strong>of</strong> Durrell's Night Gecko, <strong>the</strong> Keelscaled<br />
Tree Boa, and 33 specimens <strong>of</strong><br />
Gun<strong>the</strong>r's Gecko - exceeding <strong>the</strong> results<br />
<strong>of</strong> all previous expeditions - were<br />
observed and measured accordingly.<br />
Bouton's Skinks, in accordance with <strong>the</strong><br />
former observations, seemed to be confined<br />
to <strong>the</strong> rocky shoreline surfaces. The<br />
Mauritian nature conservation authorities<br />
put enormous efforts into <strong>the</strong><br />
restoration <strong>of</strong> <strong>the</strong> native vegetation <strong>of</strong><br />
Round Island, which could help to stabilize<br />
<strong>the</strong> different reptile populations.<br />
During <strong>the</strong> second expedition (April<br />
2001) our aim was to search for evidence<br />
<strong>of</strong> <strong>the</strong> presence or extinction <strong>of</strong><br />
<strong>the</strong> Burrowing Boa (Bolyeria multocarinata<br />
(Boie, 1827)). This animal has been<br />
seen only four times in <strong>the</strong> 20th century,<br />
and since its last observation in 1974 no<br />
signs <strong>of</strong> its survival have been discovered.<br />
Because our searches, too, were<br />
unsuccessful in this respect, we consider<br />
this snake species to be extinct.<br />
In our detailed reports (available through<br />
<strong>the</strong> National Parks and Conservation<br />
Service, Reduit, Mauritius) conclusions<br />
were drawn about <strong>the</strong> status <strong>of</strong> and <strong>the</strong><br />
possible threats to <strong>the</strong> unique herpet<strong>of</strong>aunal<br />
assemblage <strong>of</strong> Round Island. We<br />
also proposed to <strong>the</strong> Mauritian nature<br />
conservation authorities to organize<br />
more regular and standardized surveys<br />
to follow <strong>the</strong> changes in <strong>the</strong> different<br />
reptile populations.<br />
Acknowledgements<br />
We would like to extend our sincere thanks to <strong>the</strong> following persons for <strong>the</strong>ir assistance<br />
during our visit to <strong>the</strong> island: Youssef Mungroo, Vishnu Bachraz, Krishna Puttoo,<br />
Shyamduth Ramrekha, Vishal Nundlaul (all from <strong>the</strong> Ministry <strong>of</strong> Agriculture, Food,<br />
Technology & Natural Resources, National Parks and Conservation Service, Reduit,<br />
Mauritius), Carl Jones and Ashok Khadun (Mauritian Wildlife Appeal Fund, Port Louis,<br />
Mauritius). Our travel could not have been accomplished without <strong>the</strong> financial help <strong>of</strong><br />
Comp Travel Ltd. (Budapest, Hungary), <strong>the</strong> Fauna & Flora International (Great Britain),<br />
and <strong>the</strong> Mauritius Travel & Tourist Bureau Ltd. (Floreal, Mauritius). Special thanks are<br />
due to our wives, Zita Zachar & Agnes Varga, for helping and encouraging us in <strong>the</strong> field<br />
(as well as before and after <strong>the</strong> expedition).<br />
REFERENCES<br />
ARNOLD, E. N. 1980: Recently extinct reptile populations from Mauritius and Reunion,<br />
Indian Ocean. Journal <strong>of</strong> Zoology, London 191: 33-47.<br />
ARNOLD, E. N. & JONES, C. G. 1994: The night geckos <strong>of</strong> <strong>the</strong> genus Nactus in <strong>the</strong><br />
Mascarene islands with a description <strong>of</strong> <strong>the</strong> distinctive population on Round<br />
Island. Dodo, Jersey Wildlife Preservation Trust 30: 119-131.<br />
BLOXAM, Q. & VOKINS, M. 1978: Breeding and maintenance <strong>of</strong> Phelsuma guen<strong>the</strong>ri<br />
(Boulenger, 1985) at <strong>the</strong> Jersey Zoological Park. Dodo, Jersey Wildlife<br />
Preservation Trusts 15: 82-91.<br />
BULLOCK, D. J. 1977: Round Island - A tale <strong>of</strong> destruction. Oryx 14: 51-58.<br />
BULLOCK, D. J. 1986: The ecology and conservation <strong>of</strong> reptiles on Round Island and
80 Biota 3/1-2,2002 KORSOS & TROCSANYI<br />
Gunner's Quoin, Mauritius. Biological Conservation 37: 135-156.<br />
BULLOCK, D. & NORTH, S. 1975: Report <strong>of</strong> <strong>the</strong> Edinburgh University Expedition to Round<br />
Island, Mauritius, July and August 1975. Unpublished report, University <strong>of</strong><br />
Edinburgh.<br />
BULLOCK, D. & NORTH, S. 1984: Round Island in 1982. Oryx 18: 36-41.<br />
DASZAK, P. 1994: The 1993 Raleigh International Round Island Expedition, including a survey<br />
<strong>of</strong> intestinal parasites collected from animals <strong>of</strong> Round Island, Mauritius and<br />
Rodrigues and observations on Round Island reptiles. Unpublished report,<br />
Kingston University.<br />
DULLOO, M. E., BULLOCK, D. J. & NORTH, S. 1996: Report <strong>of</strong> <strong>the</strong> expedition to Round<br />
Island and Gunner's Quoin, Mauritius, July/August 1996. Unpublished report,<br />
Mauritius Wildlife Foundation.<br />
GARBUTT, N. 1992: The reptiles <strong>of</strong> Round Island, Mauritius. Herptile 17: 157-170.<br />
GARBUTT, N. 1992: The reptiles <strong>of</strong> Round Island, Mauritius. Vivarium 4: 14-18, 32, 33.<br />
GARBUTT, N. 1992: The Round Island gecko: A most unusual Phelsuma. Dactylus 1: 17-21.<br />
JONES, C. G. 1988: Round Island boa eats Serpent Island gecko. Oryx 22: 180.<br />
JONES, C. G. 1993: The ecology and conservation <strong>of</strong> Mauritian skinks. Royal Society <strong>of</strong> Arts<br />
and Sciences, Mauritius 5: 71-95.<br />
JONES, C. G. & HARTLEY, J. 1995: A conservation project on Mauritius and Rodrigues: An<br />
overview and bibliography. Dodo, Jersey Wildlife Preservation Trust 31: 40-65.<br />
KORS6S, Z. & TR6CSANYI, B. 2000: Report on <strong>the</strong> Hungarian Herpetological Expedition to<br />
Round Island, Mauritius, November 1999. Unpublished report, Hungarian<br />
Natural History Museum & University <strong>of</strong> Pecs.<br />
KORSOS, Z. & TROCSANYI, B. 2001: Population assessment <strong>of</strong> Gun<strong>the</strong>r's Gecko in its natural<br />
habitat, Round Island, Mauritius. 15th Annual Meeting <strong>of</strong> <strong>the</strong> Society <strong>of</strong><br />
Conservation Biology, University <strong>of</strong> Hawaii, Hilo, USA, July 29-August 1, 2001<br />
MERTON, D. V., ATKINSON, I. A. E., STRAHM, W, JONES, C., EMPSON, R. A.,<br />
MUNGROO, Y, DULLOO, E. & LEWIS, R. 1989: A management plan for <strong>the</strong><br />
restoration <strong>of</strong> Round Island, Mauritius. Jersey Wildlife Preservation Trust.<br />
NORTH, S. G. & BULLOCK, D. J. 1986: Changes in vegetation and populations <strong>of</strong> introduced<br />
mammals <strong>of</strong> Round Island and Gunner's Quoin, Mauritius. Biological<br />
Conservation 37: 99-117.<br />
NORTH, S. G., BULLOCK, D. J. & DULLOO, M. E. 1994: Changes in <strong>the</strong> vegetation and reptile<br />
populations on Round Island, Mauritius, following eradication <strong>of</strong> rabbits.<br />
Biological Conservation 67: 21-28<br />
OWADALLY, A. W. & LAMBERT, M. 1988: Herpetology in Mauritius. A history <strong>of</strong> extinction,<br />
future hope for conservation. Bulletin <strong>of</strong> <strong>the</strong> British Herpetological Society 23:11-<br />
20.<br />
RAMREKHA, S. 1999: Expedition on Round Island, 17th-25th August 1999. Unpublished<br />
report, National Park and Conservation Service, Mauritius.<br />
TONGE, S. 1990: The past, present and future <strong>of</strong> <strong>the</strong> herpet<strong>of</strong>auna <strong>of</strong> Mauritius. Bulletin <strong>of</strong><br />
<strong>the</strong> Chicago Herpetological Society 25: 220-226.<br />
TR6CSANYI, B. 1997: The artificial environment and activity: Some aspects <strong>of</strong> <strong>the</strong> captive<br />
maintenance <strong>of</strong> <strong>the</strong> Round Island Gecko (Phelsuma guen<strong>the</strong>ri). Thesis, University<br />
<strong>of</strong> Kent & Jersey Wildlife Preservation Trust.<br />
VINSON, J. 1953: Some recent data on <strong>the</strong> fauna <strong>of</strong> Round and Serpent Islands. Proceedings<br />
<strong>of</strong> <strong>the</strong> Royal Society <strong>of</strong> Arts and Sciences, Mauritius 1: 253-257.<br />
VINSON, J. & VINSON, J.-M. 1969: The saurian fauna <strong>of</strong> <strong>the</strong> Mascarene islands. Bulletin <strong>of</strong><br />
<strong>the</strong> Mauritius Institute 6: 203-320.<br />
VINSON, J.-M. 1975: Notes on <strong>the</strong> reptiles <strong>of</strong> Round Island. Bulletin <strong>of</strong> <strong>the</strong> Mauritius<br />
Institute 8: 49-67.
KORSOS<br />
& TROCSANYI Bio la 31-3, 2003 81<br />
2. Palm savannah: typical<br />
vegetation <strong>of</strong> <strong>the</strong><br />
western part <strong>of</strong> Round<br />
Island<br />
1. Round Island from <strong>the</strong> air<br />
3. Gun<strong>the</strong>r's Gecko<br />
Pheisuma guen<strong>the</strong>ri<br />
Boulenger, 1885
82 Biota 3/1-2, 2002 KORSOS & TROCSANYI<br />
5. Ornate Day Gecko<br />
Phelsuma ornata Gray,<br />
1825<br />
- .<br />
4. Gun<strong>the</strong>r's Gecko<br />
Phelsuma guen<strong>the</strong>ri<br />
Boulenger, 1885<br />
6. Durrell's Night<br />
Gecko Nactus serpensinsula<br />
durrelli<br />
Arnold & Jones, 1994
KORSOS & TROCSANYI BJOla 3/1-2, 2002 83<br />
8. Bojer's Skink<br />
Congylomorphus<br />
bojerii (Desjardins,<br />
1831)<br />
7. Telfair's Skink<br />
Leioiopisma telfairii<br />
(Desjardins, 1831)<br />
9. Bouton's Skink<br />
Cryptoblepharus boutonii<br />
(Desjardins,<br />
1831)
84 Biota 3/1-2, 2002 KORSOS & TROCSANYI<br />
11. Keel-scaled Tree<br />
Boa Casarea dussumieri<br />
(Schle gel, 1837).<br />
Adult's head<br />
10. Burrowing Round Island Boa Bolyeria multocarinata (Bole,<br />
1827)<br />
The only museum specimen in Mauritius<br />
12. Keel-scaled Tree<br />
Boa Casarea dussumieri<br />
(Schlegel, 1837).<br />
Juvenile colouration
KUTRUP & YILMAZ Biota 3/i-a, 2002 85<br />
Preliminary data on some new<br />
specimens <strong>of</strong> Vipera barani<br />
collected from Trabzon<br />
(Nor<strong>the</strong>astern Turkey)<br />
Bilal KUTRUP1 & Nurh ay at YILMAZ2<br />
1Karadeniz Technical University, Faculty <strong>of</strong> Arts and Sciences, Dept. <strong>of</strong> Biology, 61080,<br />
Trabzon, Turkey<br />
E-mail: kutrup@ktu.edu.tr<br />
2Karadeniz Technical University, Faculty <strong>of</strong> Rize, Arts&Sciences, Dept. <strong>of</strong> Biology, 53100,<br />
Rize, Turkey<br />
E-mail: nurhayat@ktu.edu.tr<br />
Abstract<br />
This study was carried out to find viper specimens in Trabzon, situated in nor<strong>the</strong>astern<br />
Turkey. During <strong>the</strong> study period, a total <strong>of</strong> 9 viper specimens were collected from new<br />
locations in Arpaozu (Caykara), Balhca, Sugeldi (Of), and Camlik (Vakfikebir). For morphometric<br />
studies, <strong>the</strong>se specimens were examined, and for all <strong>of</strong> <strong>the</strong>se specimens, 16<br />
different items <strong>of</strong> data were collected. In addition, collected data were compared with<br />
data from o<strong>the</strong>r vipers (Vipera barani and Vipera pontica) which are described from<br />
Rize, Artvin and Adapazan. Trabzon viper specimens are characterised by partial fragmentation<br />
<strong>of</strong> frontal and parietals, high ventrals and loreals, fewer subcaudals and yellow<br />
green tail tips than in Vipera barani and Vipera pontica. When comparing <strong>the</strong> subalpine<br />
population (Arpaozu) with o<strong>the</strong>r lowland ones (Of, Yomra and Vakfikebir), differences<br />
in apical and dorsal patterns could be detected. The Arpaozu population had<br />
one apical, which is normally found only in Vipera ursiniiand occasionally in <strong>the</strong> Vipera<br />
kaznakovi complex (V. kaznakovi, V. dinniki and V. darevski). On <strong>the</strong> o<strong>the</strong>r hand, <strong>the</strong><br />
number <strong>of</strong> apicals was 2 or 3 in Vipera barani and V. pontica. Also, <strong>the</strong> number <strong>of</strong> zigzag<br />
bands was low in <strong>the</strong> Arpaozu specimens (48-49 instead <strong>of</strong> 62-67 in <strong>the</strong> lowland<br />
specimens). As a result, it is clearly indicated that <strong>the</strong> new viper specimens from Trabzon<br />
show some similarities in colour patterns as well as scalation characters with V. barani<br />
and V. pontica. However, <strong>the</strong> number <strong>of</strong> circumoculars, loreals and crown scales is lower<br />
in our specimens than in both species. Taking into consideration <strong>the</strong> differences and<br />
similarities, <strong>the</strong> Trabzon specimens are most similar to V. barani<br />
Key words: Reptilia, Viperidae, Vipera barani, Trabzon<br />
Received 30 August; accepted 20 December 2001
86 3/1-2, 2OO2 KUTRUP & YILMAZ<br />
INTRODUCTION<br />
Recent studies <strong>of</strong> <strong>the</strong> systematics <strong>of</strong><br />
vipers in <strong>the</strong> Caucasus have shown that<br />
<strong>the</strong> taxonomy <strong>of</strong> this group is ra<strong>the</strong>r complex.<br />
It has been reported that <strong>the</strong>re are<br />
two viper groups in this region, Vipera<br />
kaznakovi and Vipera ursinii (Vedmederja<br />
et al. 1986, Nilson et al. 1995). Vipera<br />
barani has been evaluated to be an<br />
important species in <strong>the</strong> Vipera kaznakovi<br />
group as Vipera pontica. Only Vipera<br />
kaznakovi is well defined and restricted in<br />
distribution and geographically separated<br />
from <strong>the</strong> o<strong>the</strong>r species in this region.<br />
Among <strong>the</strong>se species, Vipera pontica<br />
shows clear similarities with Vipera barani<br />
in having high fragmentation <strong>of</strong> <strong>the</strong><br />
frontal and parietals, in <strong>the</strong> number <strong>of</strong><br />
ventrals as well as by <strong>the</strong>ir yellow green<br />
tail (Billing et al. 1990).<br />
Initially, Vipera barani Bohme and Joger,<br />
1983, was originally described from a single<br />
specimen - a melanistic female from<br />
Sapanca (Adapazan) which is situated on<br />
<strong>the</strong> southwest coast <strong>of</strong> <strong>the</strong> Black Sea.<br />
Then, Frazen and Heckes identified and<br />
described Vipera barani from Rize<br />
(Nilson, pers. comm.). Recently Baran et<br />
al. (1997) published information about<br />
three viper specimens that <strong>the</strong>y considered<br />
as being V. pontica from<br />
Camlyhemsin (Rize). Never<strong>the</strong>less, some<br />
German herpetologists (Frazen and<br />
Heckes) believe that <strong>the</strong> Camlyhemsin<br />
population should be evaluated as V.<br />
barani<br />
The taxonomy <strong>of</strong> <strong>the</strong> Vipera barani and<br />
Vipera pontica has been confusing and<br />
contradictory (Hoggren et al. 1993,<br />
Nilson et al. 1994, 1995, Joger et al.<br />
1997). The original question <strong>of</strong> whe<strong>the</strong>r<br />
<strong>the</strong> specimens belonging to <strong>the</strong> Eastern<br />
Black Sea Region (Rize and Trabzon) and<br />
those <strong>of</strong> Adapazan are identical must be<br />
answered. For this purpose, DNA<br />
sequences <strong>of</strong> <strong>the</strong> vipers caught from<br />
Adapazan and Rize have been studied by<br />
Frazen and Heckes. There is ano<strong>the</strong>r<br />
question; could it be that what we have is<br />
a series <strong>of</strong> populations within <strong>the</strong> subgenus<br />
pelias along <strong>the</strong> Anatolian Black<br />
Sea coast? The new specimens caught<br />
from Trabzon could fit into such a series.<br />
On <strong>the</strong> o<strong>the</strong>r hand, Camlyhemsin and<br />
Adapazan specimens were evaluated as<br />
two different groups (V. barani in berus<br />
group and V. pontica in aspisfgroup) by<br />
Joger et al. (1997). They also stated that<br />
<strong>the</strong> melanistic specimens belonging to V.<br />
barani show similarities with <strong>the</strong> aspis<br />
group. Recently, <strong>the</strong>re has been a trend<br />
to evaluate both species as <strong>the</strong> same<br />
taxon.<br />
Despite intensive searching by several<br />
herpetologists, no additional Vipera<br />
barani specimen was captured.<br />
Examining more specimens belonging to<br />
Viper from new localities in Trabzon is<br />
aimed at gaining a broad perspective<br />
including colour pattern and pholidosis<br />
characteristics.<br />
MATERIAL AND METHODS<br />
The study was conducted from April to<br />
September <strong>of</strong> 1999 and 2000, and joint<br />
field trips were made in different parts <strong>of</strong><br />
Trabzon to capture <strong>the</strong> samples <strong>of</strong> viper.<br />
A total <strong>of</strong> nine specimens (5 females and<br />
4 males) were captured from four different<br />
locations. Two adult females were<br />
caught from <strong>the</strong> locality <strong>of</strong> Arpaozu, situated<br />
approximately 20 km south <strong>of</strong><br />
Uzungol (Trabzon). This habitat was separated<br />
from <strong>the</strong> o<strong>the</strong>rs by having steep<br />
wooded mountain slopes with many big,<br />
rocky outcrops at high altitudes (more<br />
than 2000m).<br />
The o<strong>the</strong>r specimens were caught in <strong>the</strong><br />
lowland Black Sea coast locations <strong>of</strong><br />
Ballica, Sulgeldi (Of), Cinarh (Yomra) and<br />
Camhk (Vakfikebir). One adult male was<br />
captured from Ballica, situated approximately<br />
63 km east <strong>of</strong> Trabzon and ano<strong>the</strong>r<br />
adult male was encountered in Sugeldi,<br />
situated 5 km to <strong>the</strong> eastern part <strong>of</strong><br />
Ballica. One adult female was caught in
KUTRUP & YILMAZ 3/1-2, 2O02 87<br />
Cmarli, which is nearby in <strong>the</strong> east <strong>of</strong><br />
Trabzon. Two adult males, one adult and<br />
young females were retrieved from<br />
Camlik which is 68 km west <strong>of</strong> Trabzon.<br />
The locality <strong>of</strong> Cumhuriyet is approximately<br />
one km from <strong>the</strong> sea with an altitude<br />
<strong>of</strong> 80 m., while <strong>the</strong> Ballica and<br />
Sugeldi localities are two or three km<br />
from <strong>the</strong> sea, with an altitude <strong>of</strong> 250-330<br />
m. Although Ballica and Sugeldi specimens<br />
were captured near <strong>the</strong> tea plant<br />
Camellia sirennis, <strong>the</strong> Cmarli and<br />
Cumhuriyet specimens were captured on<br />
short grassland populated by Oak<br />
Quercussp., Alder Alnus sp. and hazelnut<br />
Corylus sp. trees.<br />
Pattern and coloration were examined<br />
and noted for five adult and three immature<br />
specimens. In addition, black and<br />
white slides were taken. Then specimens<br />
were fixed with 70% ethanol. For morphometric<br />
studies, 9 vipers were examined,<br />
and for all <strong>of</strong> <strong>the</strong>se specimens 16<br />
different items <strong>of</strong> data were obtained.<br />
Total and tail length were measured. In<br />
addition, <strong>the</strong> number <strong>of</strong> ventrals, subcaudals,<br />
anterior, mid-body and posterior<br />
dorsal scale rows, apical plates, supralabials,<br />
sublabials, circumocular scales, loreals,<br />
chanthals, crown scales and zig-zag<br />
spirals in <strong>the</strong> dorsal band were counted. A<br />
fur<strong>the</strong>r rostral index (height/breadth) was<br />
calculated. The division <strong>of</strong> parietals and<br />
frontals was noted. This information was<br />
used in morphological description, taxonomical<br />
analyses and comparison with<br />
<strong>the</strong> o<strong>the</strong>r Vipera barani and Vipera pontica<br />
captured from different locations.<br />
RESULTS AND DISCUSSION<br />
The new specimens from lowland populations<br />
<strong>of</strong> Yomra, Of and Vakfikebir show<br />
some differences in terms <strong>of</strong> scalation and<br />
colour pattern in comparison to <strong>the</strong> subalpine<br />
population (Arpaozu). Also, our<br />
specimens were compared to Vipera<br />
barani and Vipera pontica, which were<br />
described from Adapazan and Rize, in<br />
order to clarify <strong>the</strong>se morphological similarities<br />
and differences within both<br />
species. In addition, <strong>the</strong>y were compared<br />
to <strong>the</strong> Camlyhemsin specimens, which<br />
had not yet received a clear taxonomic<br />
position (Table 1).<br />
Table 1. External morphology <strong>of</strong> <strong>the</strong> Trabzon viper specimens collected from <strong>the</strong><br />
Arpaozu, Of, Yomra and Vakfikebir and related taxa (V. barani and V. pontica)<br />
Characters<br />
Tail length (mm)<br />
Total length (mm)<br />
Ventral<br />
Subcaudal<br />
Dorsal scales<br />
Neck<br />
Mid-body<br />
Pasterior<br />
Apicals<br />
Chantals<br />
Rostral index<br />
Loreals<br />
Supralabials<br />
Sublabiais<br />
Circumoculars<br />
Crown scales<br />
Zig-zag band<br />
Arpaozu<br />
2(2)<br />
52-73<br />
444-590<br />
142<br />
28-29<br />
25-26<br />
21<br />
17<br />
1<br />
2-2<br />
1.12-1.24<br />
4-5<br />
8-9<br />
9-11<br />
8-9<br />
18-24<br />
48(49)<br />
Of<br />
2(6")<br />
82-83<br />
518-543<br />
140-141<br />
33-37<br />
24-25<br />
21<br />
17-18<br />
2<br />
2-2<br />
1.05-1.18<br />
5-5<br />
9-9<br />
11-11<br />
10-13<br />
22-25<br />
67-*<br />
Yomra<br />
1(2)<br />
78<br />
634<br />
148<br />
29<br />
26<br />
21<br />
17<br />
2<br />
2-2<br />
1.10<br />
4-5<br />
9-8<br />
10-11<br />
11<br />
26<br />
56<br />
2(2)<br />
22-69<br />
176-627<br />
149<br />
30-31<br />
23-25<br />
21<br />
17<br />
2<br />
2-2<br />
1.05-1.07<br />
5-4<br />
8-9<br />
10-11<br />
9-10<br />
22-25<br />
62-65<br />
= Character was not visible because <strong>of</strong> damaged specimen<br />
Vakfikebir<br />
2(
88 Biota 3 -2, 2OO2 KUTRUP & YILMAZ<br />
As can be seen in Table 1, <strong>the</strong> number <strong>of</strong><br />
ventrals in <strong>the</strong> specimens caught from<br />
Trabzon falls between 141 and 145,<br />
except for two specimens collected from<br />
Yomra and Vakfikebir. These specimens<br />
have a high number <strong>of</strong> ventrals (148-<br />
149), a feature which is not counted in V.<br />
barani and V. pontica. The number <strong>of</strong><br />
subcaudals in males is higher than in<br />
females (33-38 instead <strong>of</strong> 28-31 in<br />
females). Also, it is obvious that males<br />
have longer tails than females.<br />
Consequently, <strong>the</strong> rate <strong>of</strong> tail length to<br />
total length, which is 0.11-0.13 in<br />
females and 0.15-0.17 in males, confirms<br />
this result. The Arpaozu population (two<br />
females) shows remarkable differences<br />
from <strong>the</strong> o<strong>the</strong>r lowland populations in<br />
having one apical (Figure 1), a feature<br />
which is normally found only in Vipera<br />
ursinii, and occasionally in <strong>the</strong> V. kaznakovicomplex<br />
(V. kaznakovi, V. dinniki<br />
and V. darevski). In contrast, one apical<br />
was not reported ei<strong>the</strong>r in V. barani or in<br />
Figure 1. Dorsal view <strong>of</strong> <strong>the</strong> head <strong>of</strong> <strong>the</strong><br />
adult female from Arpaozu<br />
patterns <strong>of</strong> <strong>the</strong> Arpaozu specimens are<br />
very different from <strong>the</strong> members <strong>of</strong> this<br />
group (V. kaznakovi complex).<br />
The number <strong>of</strong> crown scales (interchantals<br />
+intersupraoculars ) is much lower in<br />
Trabzon specimens (18-26) than in <strong>the</strong> V.<br />
pontica (34), V. barani (25-40) and<br />
Camlyhemsin specimens (25-33). But<br />
only a young male from Vakfikebir differs<br />
from <strong>the</strong> o<strong>the</strong>rs in having higher crown<br />
scales (33) than in <strong>the</strong> V. barani and V.<br />
pontica specimens.<br />
Additionally, <strong>the</strong> Trabzon specimens have<br />
lower loreal counts (4-5, 5-4 and 5-5)<br />
(Figure 2). Four <strong>of</strong> <strong>the</strong> specimens examined<br />
have 5-5 loreals like <strong>the</strong><br />
Camlyhemsin ones (5-5), while <strong>the</strong> o<strong>the</strong>rs<br />
show similarities with V. barani (3-6).<br />
Fur<strong>the</strong>rmore, our specimens differ from<br />
Figure 2. Lateral view <strong>of</strong> <strong>the</strong> head <strong>of</strong> <strong>the</strong><br />
adult male from Balhca<br />
<strong>the</strong> V. pontica, V. barani and<br />
Camlyhemsin specimens in having fewer<br />
circumoculars (9-11 instead <strong>of</strong> 10-14 in<br />
\. pontica V. (Table barani, 1). 11-12 However, in V. pontica colour and 10-12<br />
in <strong>the</strong> Camlyhemsin specimens) On <strong>the</strong><br />
o<strong>the</strong>r hand, <strong>the</strong> Balhca specimens show<br />
clear similarities in circumocular counts<br />
(12-13) with V. barani (figure 2).<br />
Head scales between <strong>the</strong> rostral and posterior<br />
end <strong>of</strong> <strong>the</strong> parietal area, as well as<br />
temporal, are not keeled as in V. barani<br />
and V. pontica. The head morphology <strong>of</strong><br />
<strong>the</strong> Trabzon specimens differs from that<br />
<strong>of</strong> V. pontica in having much more blunt<br />
and expanded snouts than any member
KUTRUP & YILMAZ Biola '31-2,2002 89<br />
Figure 3. Dorsal view <strong>of</strong> <strong>the</strong> adult Ballica<br />
male<br />
<strong>of</strong> <strong>the</strong> berus complex (Figure 1).<br />
According to Billing et al. (1990), V. pontica<br />
has a pronounced snout Also our<br />
specimens do not have <strong>the</strong> markedly<br />
raised snouts which are normally found in<br />
several Caucasian viper populations such<br />
as V. pontica (Nilson et al. 1995).<br />
A great number <strong>of</strong> different colour<br />
morphs was expressed in <strong>the</strong> Caucasian<br />
vipers (Billing et al. 1990, Hoggren et al.<br />
1993, Nilson et al. 1995, Kutrup 1999).<br />
Although <strong>the</strong> Arpaozu locality, which was<br />
situated in high subalpine mountain belts<br />
(2000m) such as <strong>the</strong> Vipera dinniki locality<br />
(Nilson et al. 1994, 1995, Hoggren et<br />
al. 1993), has very different climatic factors<br />
and plant species, specimens belong-<br />
ing to Arpaozu did not show any significant<br />
differences in terms <strong>of</strong> colour and<br />
pattern from <strong>the</strong> lowland specimens.<br />
Only <strong>the</strong> number <strong>of</strong> spirals in <strong>the</strong> dorsal<br />
zig-zag band was lower in <strong>the</strong> Arpaozu<br />
specimens (48-49 instead <strong>of</strong> 62-67 in <strong>the</strong><br />
Of, Yomra, and Vakfikebir specimens,<br />
respectively) (Figure 3, Table 1). This high<br />
number <strong>of</strong> zig-zag bands (67) was normally<br />
found in <strong>the</strong> high populations <strong>of</strong> V.<br />
dinniki (Nilson et al. 1995, Joger et al.<br />
1997). On <strong>the</strong> o<strong>the</strong>r hand, <strong>the</strong> number <strong>of</strong><br />
zig-zag bands which was observed in<br />
adult samples was lower than in young<br />
ones. This could be given an ontogenetic<br />
explanation, as pattern <strong>of</strong>ten fades with<br />
age (Nilson et al. 1995).<br />
Only two melanistic specimens (one<br />
female from Arpaozu and one male from<br />
Sugeldi) were observed during <strong>the</strong> study<br />
period (Figure 1). In <strong>the</strong>se samples, a<br />
black coloration with high melanin production<br />
covered all o<strong>the</strong>r colour patterns.<br />
In contrast, dorsal patterns belonging to<br />
unmelanistic specimens consisted <strong>of</strong> light<br />
brown in adults and dark brown in <strong>the</strong><br />
young, and broad, black-bordered zigzag<br />
spirals with a total <strong>of</strong> 48-67 in both<br />
adults and young. Also, many dark<br />
blotches, which were vertically placed<br />
and not connected to <strong>the</strong> zig-zag band,<br />
were observed along <strong>the</strong> body sides.<br />
Head patterns <strong>of</strong> two black bands<br />
extended from <strong>the</strong> posterior end <strong>of</strong> <strong>the</strong><br />
parietal area to <strong>the</strong> sides <strong>of</strong> <strong>the</strong> neck and<br />
<strong>the</strong>y had wide, black-bordered bands,<br />
which ran from <strong>the</strong> eyes to <strong>the</strong> neck<br />
along <strong>the</strong> upper sides <strong>of</strong> <strong>the</strong> supralabials<br />
(Figure 2). All unmelanistic specimens<br />
had a blotch which was approximately<br />
elliptical on <strong>the</strong> neck, just as in V. barani<br />
from Adapazan (Joger et al. 1997) . The<br />
edges <strong>of</strong> <strong>the</strong> supralabials, rostral , chanthals<br />
and supraoculars were white. All <strong>of</strong><br />
<strong>the</strong> samples had black ventrals. Although<br />
bigger white spots were only seen on <strong>the</strong><br />
ventral <strong>of</strong> <strong>the</strong> head in <strong>the</strong> melanistic samples,<br />
many large and small white spots
90 Biota 3/1-2,2002 KUTRUP & YILMAZ<br />
extended from mentale to mid body area<br />
in <strong>the</strong> unmelanistic samples. These white<br />
spots also ran along <strong>the</strong> sides <strong>of</strong> <strong>the</strong> ventrals<br />
to <strong>the</strong> anale in young samples. The<br />
yellow coloration, which was seen at <strong>the</strong><br />
posterior area <strong>of</strong> <strong>the</strong> subcaudals, was fairly<br />
distinct in all specimens.<br />
An examination <strong>of</strong> <strong>the</strong> morphology <strong>of</strong> <strong>the</strong><br />
different specimens clearly indicate that<br />
Trabzon vipers show some differences in<br />
terms <strong>of</strong> colour patterns as well as scalation<br />
characteristics from V. barani and V.<br />
pontica. On <strong>the</strong> o<strong>the</strong>r hand, <strong>the</strong>se vipers<br />
show similarities with V. barani and V.<br />
pontica in scalation as well as colour patterns.<br />
According to <strong>the</strong> scalation, <strong>the</strong><br />
specimens <strong>of</strong> Trabzon vipers may be<br />
belong to ei<strong>the</strong>r V. barani or V. pontica.<br />
But <strong>the</strong> colour patterns <strong>of</strong> our melanistic<br />
specimens are more similar to those in V.<br />
barani than to those in V. pontica.<br />
However, <strong>the</strong> number <strong>of</strong> crown scales<br />
(interchanthals + intersupraoculars), loreals<br />
and circumoculars is lower in our specimens<br />
than in both o<strong>the</strong>r species.<br />
Taking into account <strong>the</strong> differences and<br />
similarities as indicated above, Trabzon<br />
samples have more similarities with V.<br />
barani than with V. pontica. Ultimately,<br />
we evaluate our vipers as Vipera barani.<br />
Fur<strong>the</strong>rmore, our viper samples can be<br />
evaluated as belonging to a subspecies <strong>of</strong><br />
V. barani (Joger, pers. comm.).<br />
It must be borne in mind that we would<br />
need a larger series <strong>of</strong> molecular data to<br />
find out more.<br />
REFERENCES<br />
BARAN, I., TOSUNOGLU, M., KAYA, U. & KUMLUTAS, Y. 1997: Camlyhemsin (Rize<br />
Civarmm Herpeto- faunasi Hakkinda. Tr. j. <strong>of</strong> Zoology 21: 409-416.<br />
BILLING, H., NILSON, G. & SATTER, U. 1990: Vipera pontica sp. n., a new species in <strong>the</strong><br />
kaznakovi group (Reptia-Viperidae) from north-eastern Turkey and adjacent<br />
Transcaucasiana. Zool. Scripta 19: 231-237.<br />
B6HME, W. & JOGER, U. 1983: Eine neue Arts des Vipera berus -Komlexes aus der Turkei.<br />
Amphibia-Reptilia 4: 265-271.<br />
HOGGREN, M., NILSON, G., ANDREN, C, ORLOV, N. L & TUNIEV, B. S. 1993: Vipers <strong>of</strong><br />
<strong>the</strong> Caucasus. Natural History and Systematic Review. Herpetological Natural<br />
History 1: 11-19.<br />
JOGER, U., LENK, P., BARAN, I., BOHME, W., ZEIGLER, T, HEIDRICH, P. & WINK, M. 1997:<br />
The phylogenetic position <strong>of</strong> Vipera barani and <strong>of</strong> V. niloskii within <strong>the</strong> Vipera<br />
berus complex. Herpetologica Bonnensis: 185-194.<br />
KUTRUP, B. 1999: The Morphology <strong>of</strong> Vipera ammodytes transcaucasiana (Reptilia,<br />
Viperidae) Specimens Collected from Murgul (Artvin. Turkey), Tr. J. <strong>of</strong> Zoology<br />
23: 433-438.<br />
NILSON, G., ANDREN, C. & SZYDLAR, Z. 1994: The systematic position <strong>of</strong> <strong>the</strong> Common<br />
Adder, Vipera berus (L.) (Reptilia, Viperidae), in north Korea and adjacent regions.<br />
Bonn. Zool. Beitr. 45: 49-56.<br />
NILSON, G., TUNIEV, B., ORLOV, N. , HOGREN, M. & ANDREN, C. 1995. Systematics <strong>of</strong><br />
<strong>the</strong> Vipers <strong>of</strong> <strong>the</strong> Caucasus: Polymorphism or Sibling Species. Asiatic<br />
Herpetological Research 6: 1 -26.<br />
VEDMEDERJA, V. L., ORLOV, N. L. & TUNIEV, B. S. 1986: On <strong>the</strong> taxonomy <strong>of</strong> three viper<br />
species <strong>of</strong> <strong>the</strong> Vipera kaznakovi complex. In: Ananjeva, N. & Borkin, L. (eds).<br />
Systematics and Ecology <strong>of</strong> Amphibians and Reptiles. Proceedings <strong>of</strong> <strong>the</strong><br />
Zoological Institute, Leningrad: 55-65. In Russian.
MEESKE, SCHNEEWEISS & RYBCZYNSKI Biota 3/i-a, 2002 91<br />
Reproduction <strong>of</strong> <strong>the</strong> European<br />
Pond Turtle Emys orbicularis in<br />
<strong>the</strong> nor<strong>the</strong>rn limit<br />
<strong>of</strong> <strong>the</strong> species range<br />
A.CM. MEESKE1, N. SCHNEEWEISS2 & KJ.<br />
RYBCZYNSKi2<br />
1Zentrum fuer Naturschutz, Von-Siebold-Str.2r D-37075 Goettingen<br />
E-mail: mmeeske@gwdg.de<br />
2Naturschutzstation Rhinluch, Nauener Str. 68, D-16833 Linum<br />
E-mail: agena@t-online.de<br />
Abstract<br />
Incubation conditions and ecological requirements <strong>of</strong> nesting areas <strong>of</strong> <strong>the</strong> European<br />
Pond Turtle Emys orbicularis were investigated in <strong>the</strong> summers <strong>of</strong> 1997-2001 in one<br />
local population in southwest Lithuania. Females were trapped and each individual was<br />
weighed, measured and colourmarked. Additionally, some females were tagged with<br />
radio transmitters to find out nest sites. Nesting started between <strong>the</strong> middle <strong>of</strong> May and<br />
<strong>the</strong> first week <strong>of</strong> June. The nesting period extended over 14-20 days. Open areas on<br />
sandy dry grassland were used as nesting areas. Nest sites had expositions <strong>of</strong> 80-270°<br />
and were found on flat, slight or strongly inclined ground. The average vegetation cover<br />
amounted to 50%. 77% <strong>of</strong> females which were observed in at least three years nested<br />
in <strong>the</strong> same nesting area and 23% <strong>of</strong> females in different nesting areas. The mean nest<br />
size averaged 12,5 eggs. Females with larger body sizes frequently deposited a larger<br />
number <strong>of</strong> eggs. From captures <strong>of</strong> juveniles, successful reproduction for several years<br />
could be proved in <strong>the</strong> 90s. Suitable temperatures in summer and <strong>the</strong> following winter<br />
allow successful incubation.<br />
Keywords: Testudines, Emydidae, Emys orbicularis, reproduction, Lithuania<br />
Received 8 February2002; accepted 28 July 2002
92 Bio la 3 i-a, 2002 MEESKE, SCHNEEWEISS & RYBCZYNSKI<br />
INTRODUCTION<br />
The European Pond Turtle Emys orbicularis<br />
is a threatened species, especially in<br />
<strong>the</strong> nor<strong>the</strong>rn range <strong>of</strong> its distribution<br />
(Juszczyk 1987, Zemanek 1988, 1991,<br />
Schiemenz & Gun<strong>the</strong>r 1994, Fritz 1996,<br />
Fritz & Gun<strong>the</strong>r 1996, Fritz 2000,<br />
Schneeweiss & Fritz 2000). In particular,<br />
during <strong>the</strong> last two centuries a heavy<br />
decline <strong>of</strong> European Pond Turtle was<br />
noticed. Lithuania represents <strong>the</strong> nor<strong>the</strong>rn<br />
border <strong>of</strong> <strong>the</strong> species range (Fritz<br />
1996), where <strong>the</strong> subspecies orbicularis<br />
lives (Fritz 1992, 1998). Along this nor<strong>the</strong>rn<br />
border <strong>the</strong> climate, including short<br />
summers and cold winters, is very stressful<br />
to reproduction (Schneeweiss &<br />
Jablonski 2000).<br />
Up to <strong>the</strong> present, different sites with<br />
occurrences <strong>of</strong> turtles were found mainly<br />
in <strong>the</strong> sou<strong>the</strong>rn parts <strong>of</strong> <strong>the</strong> country<br />
(Balciauskas et al. 1997), but <strong>the</strong>re is a<br />
Figure 1. Geographic position <strong>of</strong> <strong>the</strong> study area<br />
60° N<br />
BAlClC SGA<br />
lack <strong>of</strong> information about reproduction.<br />
The actual distribution and sizes <strong>of</strong> <strong>the</strong><br />
local populations is still unknown.<br />
Never<strong>the</strong>less, young turtles were noticed<br />
in different locations in Lazdijai District in<br />
southwestern Lithuania, which is a pro<strong>of</strong><br />
<strong>of</strong> reproduction success in <strong>the</strong>se areas.<br />
The present study examines incubation<br />
conditions and ecological requirements <strong>of</strong><br />
nesting areas in <strong>the</strong> view <strong>of</strong> incubation<br />
success under natural conditions.<br />
STUDY AREA<br />
The study area is located in southwestern<br />
Lithuania near <strong>the</strong> nor<strong>the</strong>astern border<br />
with Poland (40 km SSW Alytus, Lazdijai<br />
District; 23°90'E, 54°40'N) (see Figure 1).<br />
This area experiences a cold continental<br />
climate, including warm summers and<br />
cold winters (Pearce & Smith 1993). The<br />
area contains different kinds <strong>of</strong> water<br />
bodies, partly seasonally flooded wet-
MEESKE,<br />
SCHNEEWEISS & RYBCZYNSKI Biota 3/i-a, 2002 93<br />
Figure 2. Map <strong>of</strong> <strong>the</strong> study area<br />
lands and sandy, dry areas, deciduous<br />
forests, pine forests and extensively used<br />
agricultured land. The local population<br />
inhabits two main ponds (A, G) and also<br />
uses several smaller and mainly temporary<br />
water bodies (B, C, D, E, F, H, I, K,<br />
M). Four nesting areas (I-IV) are situated<br />
in front <strong>of</strong> pine forests (see fig. 2).<br />
MATERIAL AND METHODS<br />
Before <strong>the</strong> start <strong>of</strong> <strong>the</strong> nesting period,<br />
females were captured in aquatic baited<br />
traps (method by Servan 1986) and in<br />
land traps and were weighed, measured<br />
(method by Fritz 1989, 1992, 1995) and<br />
colourmarked for identification. Some<br />
females were tagged with radio transmitters<br />
glued on <strong>the</strong> carapace. Localizations<br />
<strong>of</strong> turtles using a receiver (Stabo XR 100)<br />
connected to a hand operated unidirec-<br />
nesting area<br />
field<br />
way<br />
border <strong>of</strong> reserve<br />
tional antenna were recorded to find out<br />
nesting sites. Females were located by<br />
walking in <strong>the</strong> direction indicated by <strong>the</strong><br />
antenna (Homing-in-on-<strong>the</strong>-Animal)<br />
(White & Garrott 1990). During <strong>the</strong> nesting<br />
period <strong>the</strong> nesting areas were controlled<br />
every afternoon and evening to<br />
notice <strong>the</strong> number <strong>of</strong> females, <strong>the</strong>ir nesting<br />
behaviour and <strong>the</strong> nest locations.<br />
The degree <strong>of</strong> vegetation cover was estimated<br />
(1m2 around <strong>the</strong> nest). Expositions<br />
were noticed with a compass and inclinations<br />
with a protractor.<br />
RESULTS<br />
97% <strong>of</strong> <strong>the</strong> females (30 <strong>of</strong> 31) were<br />
mature and 94% <strong>of</strong> <strong>the</strong> females (29 <strong>of</strong> 31)<br />
were observed nesting at least once.<br />
The body sizes and body masses <strong>of</strong> mature<br />
females turtles are shown in Table 1.
94 3/1-2, 2OO2 MEESKE, SCHNEEWEISS & RYBCZYNSKI<br />
Figure 3. During nesting period 2000 daily observations <strong>of</strong> females and <strong>the</strong>ir nests in<br />
<strong>the</strong> investigation area in comparison with <strong>the</strong> daily maximum air temperature<br />
Date<br />
Beginning, duration and course <strong>of</strong> oviposition<br />
period<br />
In four years (1997r 1999-2001) <strong>the</strong><br />
nesting season started between <strong>the</strong> middle<br />
<strong>of</strong> May and <strong>the</strong> first week <strong>of</strong> June and<br />
continued 14-20 days.<br />
On various days during <strong>the</strong> nesting period<br />
<strong>the</strong> number <strong>of</strong> females wandering in<br />
<strong>the</strong> investigation area was higher than<br />
<strong>the</strong> number <strong>of</strong> observed ovipositions (see<br />
Figure 3). Some females could not be<br />
observed nesting in <strong>the</strong> known nesting<br />
areas. O<strong>the</strong>r females did not nest <strong>the</strong> first<br />
day <strong>of</strong> emergence from <strong>the</strong> water body.<br />
Table 1. Body sizes and body masses <strong>of</strong> mature females<br />
23<br />
26<br />
24<br />
22<br />
20<br />
18 ?<br />
• 16 £<br />
14 g<br />
12 |<br />
10 £<br />
S<br />
6<br />
4<br />
2<br />
131 observed Females<br />
E23 observed Nests<br />
Maximum Air Temperature °c<br />
Nesting Site Fidelity<br />
Animals inhabiting home pond A mainly<br />
used nesting areas II and III, whereas turtles<br />
inhabiting home pond G preferred<br />
nesting areas I and IV and unknown<br />
areas. Female turtles showed nesting site<br />
fidelity. 77% <strong>of</strong> females observed over<br />
three years (10 <strong>of</strong> 13) nested in <strong>the</strong> same<br />
nesting area (nesting site fidelity within<br />
300 m). 31% <strong>of</strong> <strong>the</strong> females (4 <strong>of</strong> 13)<br />
showed fidelity within 20 m. 23% <strong>of</strong> <strong>the</strong><br />
females (3 <strong>of</strong> 13) changed <strong>the</strong>ir nesting<br />
area.<br />
During five observation years, female<br />
Mean Range SD n<br />
Carapace length 170mm 146-189 mm 10.19 30<br />
Carapace width 136mm 120-152 mm 8.064 30<br />
Carapace height 72mm 54-86 nun 5.69 30<br />
Body mass 916 g 630-1252 g 365.1 25
MEESKE, SCHNEEWEISS & RYBCZYNSKI Biota 3/1-2,2002 95<br />
Table 2. Variation <strong>of</strong> nest sizes <strong>of</strong> female turtles with at least two nests in different years.<br />
Female<br />
KUC-22<br />
KUC-5<br />
KUC-12<br />
KUC-3<br />
KUC-6<br />
KUC-9<br />
KUC-16<br />
KUC-34<br />
KUC-33<br />
KUC-10<br />
KUC-17<br />
KUC-4<br />
KUC-21<br />
KUC-23<br />
Length <strong>of</strong><br />
carapace (mm)<br />
156<br />
160<br />
163<br />
169<br />
170<br />
173<br />
176<br />
176<br />
177<br />
181<br />
181<br />
182<br />
187<br />
188<br />
1997<br />
8<br />
13<br />
12<br />
11<br />
-<br />
12<br />
?<br />
-<br />
-<br />
13<br />
12<br />
9<br />
20<br />
12<br />
Number o<br />
1998<br />
-<br />
?<br />
10<br />
13<br />
9<br />
11<br />
-<br />
9<br />
12<br />
15<br />
11<br />
14<br />
16<br />
-<br />
"eggs laid<br />
1999<br />
-<br />
-<br />
9 9<br />
Figure 4. Comparison <strong>of</strong> females' carapace sizes with <strong>the</strong>ir clutch sizes in<br />
four observation years<br />
155 160 165 170 175 180 185 , 190 r 195 „<br />
Carapaxlength (mm)<br />
KUC-17 nested in three different nesting<br />
areas (nesting area I: 1997/98; nesting<br />
area II: 1999/2000; nesting area III:<br />
2001). Nesting area II included most <strong>of</strong> all<br />
nests. Before and after <strong>the</strong> first change <strong>of</strong><br />
<strong>the</strong> nesting area, KUC-17 showed nesting<br />
site fidelity and laid eggs within 10 m in<br />
two subsequent years. Female KUC-12<br />
searched every year for <strong>the</strong> same nesting<br />
area (III), which is divided into two suitable<br />
main parts. KUC-12 regularly<br />
9<br />
9<br />
14<br />
16<br />
10<br />
-<br />
12<br />
14<br />
17<br />
?<br />
2000<br />
10<br />
11<br />
12<br />
13<br />
11<br />
13<br />
15<br />
17<br />
-<br />
-<br />
11<br />
14<br />
-<br />
15<br />
• Number <strong>of</strong> laid Eggs 1997<br />
- Number <strong>of</strong> laid Eggs 1998<br />
" Number <strong>of</strong> laid Eggs 1999<br />
Number <strong>of</strong> laid Eaas 2000<br />
changed <strong>the</strong>se locations and consequently<br />
used part I in 1997, 1999 and 2001<br />
and part II in 1998 and 2000.<br />
Clutch size<br />
The females deposited an average <strong>of</strong> 12,5<br />
eggs (range = 7-20, SE = 2,59, n = 46).<br />
Larger females frequently laid a larger<br />
number <strong>of</strong> eggs (see figure 4). Variation<br />
<strong>of</strong> nest sizes within individuals in different<br />
years is shown in Table 2.
96 Biota 3/1-2,2002 MEESKE, SCHNEEWEISS & RYBCZYNSKI<br />
Nesting females were older than 25<br />
years, up to very old females for which<br />
age was not determinable. The smallest<br />
females had <strong>the</strong> smallest clutch sizes<br />
(female KUC-22: 8 eggs in 1997; female<br />
KUC-19: 7 eggs in 2000), while one <strong>of</strong><br />
<strong>the</strong> largest and oldest females, KUC-21,<br />
laid <strong>the</strong> maximum number <strong>of</strong> 20 eggs in<br />
1997 (see Table 2).<br />
Nest site locations<br />
The females nested in open areas on<br />
spongy sandy ground on sandy dry grassland<br />
(n = 57) and in five cases on loamy<br />
ground. Around <strong>the</strong> nests, vegetation<br />
was found which showed sunny and dry<br />
conditions or sandy ground for this area.<br />
Index species are Hieradum pilosella,<br />
Artemisia scoparia and Sedum acre. The<br />
degree <strong>of</strong> vegetation cover ranged<br />
between 30-90% (mean = 55%, n = 16).<br />
The females chose places with inclinations<br />
between 0-20°. 45% <strong>of</strong> <strong>the</strong> nests<br />
(14 <strong>of</strong> 31) were on flat ground (0-5°),<br />
19% (6 <strong>of</strong> 31) on slight inclined (5-10°),<br />
and 36% (11 <strong>of</strong> 31) on strongly inclined<br />
ground (10-20°).<br />
The expositions <strong>of</strong> 50 nest sites amounted<br />
to 80-270°.<br />
Reproduction success<br />
Young turtles from different years in <strong>the</strong><br />
90s were recorded in <strong>the</strong> study area. They<br />
hatched before 1997 or after <strong>the</strong> summer<br />
<strong>of</strong> 1999. These results in successful incubation<br />
were noticed only for <strong>the</strong> nests<br />
Table 3. Results <strong>of</strong> incubation success for nests without predation from August. All nests<br />
were protected against predators, controls <strong>of</strong> nests in May 2000 (*nest controlling in<br />
October 1999).<br />
Female<br />
KUC-6*<br />
KUC-12<br />
KUC-14<br />
KUC-16<br />
KUC-20<br />
KUC-23<br />
KUC-24<br />
Clutch size<br />
9<br />
12<br />
>7<br />
14<br />
12<br />
?<br />
?<br />
Unfertilized<br />
eggs%<br />
44<br />
0<br />
0<br />
0<br />
0<br />
0<br />
1 egg<br />
Dead er<br />
before<br />
hatching<br />
0<br />
0<br />
0<br />
0<br />
0<br />
0<br />
1 embryo<br />
from 1999 during 1997-2000.<br />
After <strong>the</strong> warm summer in 1999, some<br />
nests showed signs <strong>of</strong> hatching in<br />
September and at <strong>the</strong> beginning <strong>of</strong><br />
October, but most <strong>of</strong> <strong>the</strong> juveniles left<br />
<strong>the</strong> nests during <strong>the</strong> following spring <strong>of</strong><br />
2000. 7 nests were checked after hatching<br />
and showed a high hatching rate up<br />
to 100% (see Table 3).<br />
8 nests from 1997 were dug out in <strong>the</strong><br />
middle <strong>of</strong> September during possible<br />
hatching time. Only 1 juvenile <strong>of</strong> a controlled<br />
79 eggs success<strong>full</strong>y left <strong>the</strong> egg<br />
shell before digging. Nests from 1998<br />
were not checked. Generally, no hatch-<br />
ibryo%<br />
after hatching<br />
11-22<br />
0<br />
0<br />
0<br />
8<br />
0<br />
0<br />
Remarks<br />
33% hatched success<strong>full</strong>y<br />
100% hatched success<strong>full</strong>y<br />
100% hatched success<strong>full</strong>y<br />
100% hatched success<strong>full</strong>y<br />
92% hatched success<strong>full</strong>y<br />
100% hatched success<strong>full</strong>y<br />
> 70% hatched success<strong>full</strong>y<br />
ing was observed. The next spring after<br />
<strong>the</strong> snow melted, 60% <strong>of</strong> <strong>the</strong> nests were<br />
destroyed by predators. 7 nests from <strong>the</strong><br />
year 2000 (controlled in 2001) contained<br />
78% fertilized eggs (range: 30-100%),<br />
but no hatchling left <strong>the</strong> egg shell or <strong>the</strong><br />
breeding chamber respectively. The<br />
young turtles died before hatching in<br />
summer and autumn or during winter.<br />
DISCUSSION<br />
Beginning and duration <strong>of</strong> oviposition<br />
period<br />
Studies in central Poland showed that<br />
wea<strong>the</strong>r conditions influence <strong>the</strong> onset
MEESKE, SCHNEEWEISS & RYBCZYNSKI Biota 3/1-2,2002 97<br />
<strong>of</strong> <strong>the</strong> nesting period (Zemanek & Mitrus<br />
1997) and <strong>the</strong> duration <strong>of</strong> <strong>the</strong> nesting<br />
period (Zemanek 1988)r which explains<br />
<strong>the</strong> differences in beginning and duration<br />
<strong>of</strong> nesting periods in Lithuania.<br />
Generally, females <strong>of</strong> <strong>the</strong> European Pond<br />
Turtle deposited eggs in East Germany<br />
(Breitenstein 1973, Andreas & Paul 1998,<br />
Schneeweiss et al. 1998) and in Poland<br />
(Jablonski 1992, Jablonski & Jablonska<br />
1998, Zemanek & Mitrus 1997, Mitrus &<br />
Zemanek 1998, 2000) at <strong>the</strong> end <strong>of</strong> May<br />
and in June. The results <strong>of</strong> this study support<br />
<strong>the</strong>se findings. The short summer in<br />
<strong>the</strong> nor<strong>the</strong>rn ranges does not allow later<br />
nesting (e.g. in July), because <strong>the</strong> time for<br />
incubation becomes too short. In sou<strong>the</strong>rn<br />
regions, nesting in July is not unusual<br />
(Fritz & Gun<strong>the</strong>r 1996).<br />
Nesting Site Fidelity<br />
77% <strong>of</strong> <strong>the</strong> females had nests within <<br />
300 m distance from <strong>the</strong> previous nest.<br />
31 % <strong>of</strong> <strong>the</strong> females nested within 20 m.<br />
23% <strong>of</strong> <strong>the</strong> females searched for different<br />
nesting areas, but presumably, more<br />
specimens shifted nesting areas. In two<br />
subsequent years, female KUC-10<br />
deposited eggs in <strong>the</strong> same nesting area.<br />
In <strong>the</strong> third year KUC-10 showed searching<br />
behaviour in <strong>the</strong> same area but was<br />
not observed nesting <strong>the</strong>re.<br />
Several Lithuanian females showed nesting<br />
site fidelity. Nesting site fidelity for<br />
European Pond Turtle is mentioned by<br />
Wermuth (1952) and Schneeweiss &<br />
Steinhauer (1998) for Germany, by<br />
Jablonski & Jablonska (1998) and Mitrus<br />
& Zemanek (2000) for Poland, and by<br />
Rovero & Chelazzi (1996) for Italy,<br />
although <strong>the</strong>re are no standardized definitions<br />
and no figures about nesting site<br />
fidelity. In Austria, European Pond Turtle<br />
females laid eggs within 14.5 m in 2 years<br />
(Roessler 2000). Various females <strong>of</strong><br />
Chrysemys picta marginata in Canada<br />
laid eggs within 10 m <strong>of</strong> <strong>the</strong>ir previous<br />
nest (Christens & Bider 1987). Congdon<br />
et al. (1983) investigated an inter-annual<br />
fidelity <strong>of</strong> 73% <strong>of</strong> Emydoidea blandingi<br />
females to <strong>the</strong> nest area in Michigan and<br />
Loncke & Obhard (1977) <strong>of</strong> 92% <strong>of</strong><br />
Chelydra serpentina females in Canada.<br />
The Lithuanian results are approximately<br />
coincident with <strong>the</strong> observations <strong>of</strong> o<strong>the</strong>r<br />
species.<br />
During five years <strong>of</strong> observation, female<br />
KUC-17 nested in three different nesting<br />
areas. The substrate in <strong>the</strong> first nesting<br />
area (I) seemed to be very unsuitable<br />
because <strong>of</strong> <strong>the</strong> very hard loamy ground.<br />
The reason for her next change from a<br />
suitable nesting area (II) to ano<strong>the</strong>r nesting<br />
area (III) is unknown. Regular shifting<br />
<strong>of</strong> nesting areas could be a kind <strong>of</strong> reproduction<br />
strategy in comparison with nesting<br />
site fidelity. In various nesting areas<br />
different predation pressure on nests or<br />
different incubation conditions could<br />
occur. The shifting strategy also helps in<br />
searching for new nesting areas if old<br />
places were disturbed by overgrowth <strong>of</strong><br />
bushes and trees or were destroyed by<br />
man. The known nesting areas <strong>of</strong> <strong>the</strong> 80s<br />
in <strong>the</strong> same study area are partly unsuitable<br />
because <strong>of</strong> overgrowth today<br />
(Gruodis, verbal information, 2001), so<br />
<strong>the</strong> turtles were forced to search for o<strong>the</strong>r<br />
places. Schneeweiss & Steinhauer (1998)<br />
and Mitrus & Zemanek (2000) also<br />
assume that females change nesting<br />
areas when old places are destroyed. But<br />
Schneeweiss & Steinhauer (1998) also<br />
observed that if suitable places were not<br />
on females' migration routes, <strong>the</strong> females<br />
used unsuitable places.<br />
Clutch size<br />
In Lithuania <strong>the</strong> mean clutch size averaged<br />
12.5 eggs (range: 7-20 eggs) over 4<br />
years and was independent <strong>of</strong> <strong>the</strong> beginning<br />
or <strong>the</strong> duration <strong>of</strong> nesting periods. In<br />
East Germany Etnys orbicularis females<br />
deposited an average number <strong>of</strong> 12.7<br />
eggs (Schneeweiss et al. 1998). This<br />
result agrees with <strong>the</strong> Lithuanian data.
98 Biota 3/1-2,2002 MEESKE, SCHNEEWEISS & RYBCZYNSKI<br />
Roessler (2000) found average clutch<br />
sizes <strong>of</strong> 12.4 eggs in Austria, but <strong>the</strong>re<br />
some females nested twice a year. The<br />
result <strong>of</strong> <strong>the</strong> Lithuanian range <strong>of</strong> clutch<br />
sizes is similar to observations <strong>of</strong> clutch<br />
sizes <strong>of</strong> 10-18 eggs in East Germany<br />
(Andreas & Paul 1998) and in Poland<br />
(Jablonski 1992, Zemanek & Mitrus<br />
1997, Mitrus & Zemanek 1998).<br />
Nest site locations<br />
In Lithuania similar results for nest site<br />
locations were found in comparison with<br />
o<strong>the</strong>r regions. The Lithuanian females<br />
mainly used open areas with sandy dry<br />
grassland for nesting (57 <strong>of</strong> 62 nests). In<br />
East Germany 18 <strong>of</strong> 32 nests were on<br />
xero<strong>the</strong>rmic sandy grassland and all nests<br />
were on sandy or sandy-loamy areas<br />
(Schneeweiss et al. 1998). In <strong>the</strong> study<br />
area <strong>the</strong> vegetation around <strong>the</strong> nests<br />
describes typical characteristics for nesting<br />
areas <strong>of</strong> European Pond Turtle<br />
(Meeske 1997). The degree <strong>of</strong> vegetation<br />
cover ranged between 30-90% (mean =<br />
55%, n = 16). A vegetation cover <strong>of</strong> nest<br />
sites <strong>of</strong> between 5-80% was observed in<br />
East Germany (Schneeweiss et al. 1998)<br />
and in Austria between 80-90% (Roessler<br />
2000). In Lithuania, in cases <strong>of</strong> high vegetation<br />
cover <strong>the</strong> plants were mainly<br />
species with short growth which do not<br />
impair <strong>the</strong> sun's radiation on <strong>the</strong> nest.<br />
The ecological requirements <strong>of</strong> nesting<br />
areas do not clearly differ in <strong>the</strong> nor<strong>the</strong>rn<br />
species range.<br />
Isberg (1929) mentions that <strong>the</strong> substrate<br />
type is an important incubation factor. If<br />
<strong>the</strong>re is a spongy substrate as in<br />
Lithuania, <strong>the</strong> heat <strong>of</strong> sunshine can better<br />
permeate through upper strata and<br />
humidity can ooze away or evaporate<br />
easily.<br />
The factors <strong>of</strong> solar exposure and inclination<br />
have a strong impact on <strong>the</strong> vegetation<br />
and consequently on <strong>the</strong> ecoclimate<br />
(Schaefer 1992). The nests were in places<br />
with flat, slight or strongly inclined<br />
ground. The degree <strong>of</strong> slope has an effect<br />
on <strong>the</strong> intensity <strong>of</strong> radiation. Additionally,<br />
sloping areas have <strong>the</strong> advantage that<br />
longer rain periods will do little harm to<br />
<strong>the</strong> clutches.<br />
Reproduction success<br />
In spring 2000 most <strong>of</strong> <strong>the</strong> young turtles<br />
left <strong>the</strong> nests from 1999. In Lithuania, <strong>the</strong><br />
main hatching time is in spring, as in<br />
o<strong>the</strong>r nor<strong>the</strong>rn regions where European<br />
Pond Turtle lives (East Germany: Andreas<br />
& Paul 1998; Poland: Zemanek 1992,<br />
Zemanek & Mitrus 1997, Mitrus &<br />
Zemanek 1998, 2000). After melting <strong>of</strong><br />
<strong>the</strong> snow in spring, <strong>the</strong> situation <strong>of</strong> water<br />
bodies in size and number is clearly more<br />
favourable than in autumn. The hatchlings<br />
have shorter ways to <strong>the</strong> next water<br />
body and can choose between various<br />
kinds <strong>of</strong> water bodies with diverse depth,<br />
temperature, hiding sites and food selection.<br />
Fritz & Giin<strong>the</strong>r (1996) describe<br />
"spring hatching" in comparison with<br />
"autumn hatching" as a second kind <strong>of</strong><br />
reproduction strategy, because after leaving<br />
<strong>the</strong> nests, juveniles have a long period<br />
<strong>of</strong> growth before hibernating.<br />
In different years in <strong>the</strong> 90s juveniles<br />
hatched in Lithuania, but during <strong>the</strong><br />
study only <strong>the</strong> warm summer 1999 and<br />
<strong>the</strong> gentle winter 1999/2000 guaranteed<br />
successful reproduction. Consequently,<br />
reproduction success strongly depends on<br />
wea<strong>the</strong>r conditions. The results <strong>of</strong> this<br />
study support <strong>the</strong> findings from nor<strong>the</strong>ast<br />
Germany and eastern Poland that in <strong>the</strong><br />
nor<strong>the</strong>rn species range, favourable climatic<br />
conditions in <strong>the</strong> summer and <strong>the</strong><br />
following winter are necessary for reproduction<br />
(Schneeweiss & Jablonski 2000).<br />
In <strong>the</strong> nor<strong>the</strong>rn range successful reproduction<br />
does not happen every year.<br />
That <strong>the</strong> actual reproduction success is<br />
sufficient for a survival <strong>of</strong> <strong>the</strong>se populations<br />
in Lithuania may become clear in<br />
ano<strong>the</strong>r study.
MEESKE, SCHNEEWEISS & RYBCZYNSKI Biota 3/1-2,2002 99<br />
Acknowledgements<br />
Thanks to all people, institutes, and organisations who helped to render <strong>the</strong>se investigations<br />
possible, especially Dr. Linas Balciauskas (Vilnius), Dr. Uwe Fritz (Dresden), Dr.<br />
Pranas Mierauskas (Vilnius), Pr<strong>of</strong>. Dr. Miihlenberg (Goettingen), Richard Podloucky<br />
(Hildesheim), Dr. Arunas Pranaitis (Meteliai), and to DAAD (German Academic<br />
Exchange Service), DGHT (German Society for Herpetology), Institute <strong>of</strong> Ecology<br />
(University Vilnius), Meteliai Regional Pare Service, Ministry <strong>of</strong> Environment (Vilnius),<br />
Universitaetsbund Goettingen, Center for Nature Conservation (University Goettingen).<br />
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Srodkowej w warunkach naturalnych. Przeglad Zoologiczny XXXII: 405-417.<br />
ZEMANEK, A/I. 1991: Wystepowanie zolwia blotnego, Emys orbicularis (L.) w Polsce i<br />
zagadnienia jego ochrony. Przyglad Zoologiczny. XXXV: 337-347.<br />
ZEMANEK, M. & MITRUS, S. 1997: Biologia i ochrona Zollwia blotnego Emys orbicularis w<br />
Wojewodztwie radomskim. Chronmy Przyrode Ojczysta 53/1: 67-83.
AAENEGON & SALVIDIO Biota 3/1-2,2002 103<br />
Notes on habitat, egg-laying and<br />
first record <strong>of</strong> <strong>the</strong> advertisement<br />
call <strong>of</strong> Hyperolius kihangensis<br />
Michele MENEGON1 & Sebastiano<br />
SALVIDIO2<br />
'Museo Tridentino di Storia Naturale, Via Calepina 14, C. P. 393, 1-38100 Trento,<br />
Italia,<br />
2Dipartimento per lo Studio del Territorio e delle sue Risorse (DIP.TE.RIS), Corso<br />
Europa26, Genova, 1-16132 Italia.<br />
Abstract<br />
Hyperolius kihangensis is a small treefrog endemic to <strong>the</strong> Udzungwa Scarp Forest <strong>of</strong> SE<br />
Tanzania. It breeds only in a restricted area characterised by flooded swamps with shallow<br />
water within dense highland forests. During a herpetological survey conducted in<br />
1998-1999, some ecological data were obtained and <strong>the</strong> male advertisement call was<br />
recorded at <strong>the</strong> type locality. The call <strong>of</strong> H. kihangensis is composed <strong>of</strong> a series <strong>of</strong> four<br />
discrete squeaks <strong>of</strong> low amplitude, with <strong>the</strong> fundamental frequency <strong>of</strong> <strong>the</strong> note at about<br />
2,72 kHz, and with a dominant frequency <strong>of</strong> about 3.4 kHz. The advertisement call <strong>of</strong><br />
H. kihangensis is unusual within <strong>the</strong> genus because <strong>of</strong> its peculiar tonal quality. A few<br />
Hyperolius species show similar features in <strong>the</strong>ir breeding call, for example, H. kachalo-<br />
/aefrom Zambia and H. viridigulosus fram West Africa.<br />
Key words: advertisement call, Eastern Arc forests, Hyperolius kihangensis, Tanzania<br />
Received 3 September 2001; accepted 1 April 2002
104 Biota 3/i-a, 2002 MENEGON & SALVIDIO<br />
INTRODUCTION<br />
Hyperolius kihangensis has recently been<br />
described in a restricted area in<br />
Udzungwa Scarp Forest Reserve, Eastern<br />
Arc Mountains, Tanzania (preliminarily<br />
described by Schi0tz & Westergaard in<br />
Schi0tz 1999, <strong>the</strong>n formally by Schi0tz &<br />
Westergaard 2000). This endemic<br />
treefrog is small, <strong>the</strong> male reaching 19<br />
mm, and <strong>the</strong> female 26 mm in total<br />
length. A brown hourglass (i.e. eightshaped)<br />
dorsal spot characterizes its dorsal<br />
colouration in both sexes (Schi0tz<br />
1999, Schi0tz & Westergaard 2000).<br />
Males have granular dorsal skin and a<br />
gular flap. Swollen white spots, consisting<br />
mainly <strong>of</strong> iridophores, are also present in<br />
a consistent proportion (i.e. about 25%)<br />
<strong>of</strong> <strong>the</strong> population, both in males and<br />
females (Westergaard et al. 2000).<br />
Little is known about <strong>the</strong> biology <strong>of</strong> this<br />
species (Schi0tz & Westergaard in Schi0tz<br />
1999) and <strong>the</strong> breeding call <strong>of</strong> <strong>the</strong> male<br />
has not been recorded or described.<br />
During two field expeditions in <strong>the</strong><br />
Udzungwa Scarp Forest Reserve, conducted<br />
between December 1998 and<br />
January 1999, we had <strong>the</strong> opportunity to<br />
collect more specimens <strong>of</strong> this treefrog, to<br />
observe egg clutches, and to record its<br />
call directly in <strong>the</strong> field. The aim <strong>of</strong> this<br />
paper is to describe some aspects <strong>of</strong> <strong>the</strong><br />
reproductive behaviour and <strong>the</strong> advertisement<br />
call <strong>of</strong> this little known<br />
Tanzanian treefrog.<br />
MATERIAL AND METHODS<br />
Thirty days <strong>of</strong> field search were conducted<br />
in Udzungwa Scarp Forest Reserve,<br />
Iringa Region, between December 1998<br />
and January 1999. The site's geographical<br />
coordinates were obtained with a<br />
Magellan 3000 global position system<br />
(GPS) navigator.<br />
The advertisement call <strong>of</strong> Hyperolius<br />
kihangensis was recorded with a Sony<br />
"TCM 1000a" tape recorder and a Shure<br />
prologue directional microphone. Sound<br />
analysis was carried out on an Apple<br />
Macintosh G4 personal computer. The<br />
signal was sampled at 44.100 Hertz and<br />
16 bit resolution. Canary 1.2.4 s<strong>of</strong>tware<br />
(Charif et al. 1993) was used for <strong>the</strong><br />
measurement <strong>of</strong> temporal and spectral<br />
parameters <strong>of</strong> <strong>the</strong> call, and to generate<br />
audiospectrograms. Frequency information<br />
was obtained through Fast Fourier<br />
Transform (FFT size: 4096 pt.). The call<br />
variables analysed were note duration,<br />
internote duration, fundamental frequency,<br />
dominant frequency, and changes in<br />
dominant frequency. For each parameter<br />
mean, standard deviation and range are<br />
given.<br />
RESULTS<br />
Habitat<br />
The study site is located in <strong>the</strong> Udzungwa<br />
Scarp at about 1780 m a.s.l (08°22'24"S,<br />
35°58'34"E) and is characterised by a<br />
typical highland vegetation (Moreau<br />
1935). As reported by Schi0tz (1999) and<br />
Schi0tz & Westergaard (2000),<br />
Hyperolius kihangensis was found only in<br />
a small swampy area inside <strong>the</strong> forest in<br />
proximity to <strong>the</strong> Kihanga stream. During<br />
our field research about 30 specimens <strong>of</strong><br />
both sexes were observed, and 9 (7 males<br />
and 2 females) were collected. Treefrogs<br />
were found on leaves between 30 and<br />
about 200 cm above <strong>the</strong> ground. The<br />
small swamp where <strong>the</strong> frogs were found<br />
covered a surface <strong>of</strong> about 1500 square<br />
metres. This area is more open than <strong>the</strong><br />
surrounding canopy forest, and is completely<br />
covered by grasses and shrubs.<br />
During <strong>the</strong> day <strong>the</strong> swamp received a<br />
greater quantity <strong>of</strong> sunlight than <strong>the</strong><br />
neighbouring shaded areas, and some<br />
random air temperature measurements<br />
showed relatively higher mean temperatures<br />
in <strong>the</strong> swamp (27.1 °C) in comparison<br />
with <strong>the</strong> surrounding forest (25°C).<br />
Several specimens, mating pairs and egg<br />
clutches, <strong>of</strong> which two contained tadpoles,<br />
were observed. Two egg clutches
MENEGON & SALVIDIO Biota 3/i-a, 2002 105<br />
were adherent to leaves and grasses<br />
above or not far from shallow, stagnant<br />
water and contained 47 to 76 eggs,<br />
respectively. The eggs had a black and a<br />
white pole and <strong>the</strong> slime was clear.<br />
During <strong>the</strong> day, Hyperolius kihangensis<br />
remained on branches or leaves in shade<br />
with <strong>the</strong> limbs close to <strong>the</strong> body; fingers<br />
and toes were kept between <strong>the</strong> body<br />
and <strong>the</strong> substrate. This diurnal sleeping<br />
posture is functional in reducing <strong>the</strong> surface<br />
area exposed to <strong>the</strong> air and <strong>the</strong>reby<br />
<strong>the</strong> evaporative water loss (Duellman &<br />
Trueb 1986).<br />
Six treefrog species have been listed by<br />
Schi0tz & Westergaard (2000) for this<br />
forest. These are: Hyperolius kihangensis,<br />
Hyperolius puncticulatus, Hyperolius<br />
spinigularis, Afrixalus uluguruensis,<br />
Leptopelis parkeri and Leptopelis barbouri.<br />
During our field researches we<br />
observed all <strong>of</strong> <strong>the</strong>se species, and in addition<br />
two specimens <strong>of</strong> Leptopelis uluguruensis<br />
were collected, thus confirming<br />
<strong>the</strong> previous doubtful record for <strong>the</strong><br />
Udzungwa Mountains (Schi0tz 1999).<br />
Description <strong>of</strong> <strong>the</strong> advertisement<br />
call<br />
During <strong>the</strong> month in <strong>the</strong> field, only once<br />
did we hear <strong>the</strong> breeding call <strong>of</strong> H.<br />
kihangensis, in spite <strong>of</strong> several hours <strong>of</strong><br />
night listening. The call <strong>of</strong> a Hyperolius<br />
kihangensis male (SVL 18.2 mm) was<br />
recorded after almost an hour <strong>of</strong> attempts<br />
during <strong>the</strong> night (January 2nd 1999, 9.45<br />
p.m., air temperature = 19,7°C, relative<br />
air humidity 97%). Just two sequences <strong>of</strong><br />
four notes were recorded from this<br />
voucher specimen that is now conserved<br />
in <strong>the</strong> herpetological collection <strong>of</strong> <strong>the</strong><br />
Museo Tridentino di Scienze Natural! <strong>of</strong><br />
Trento (collection number MTSN 086TA),<br />
as well as all original pictures regarding<br />
<strong>the</strong> species.<br />
The advertisement call <strong>of</strong> <strong>the</strong> male was a<br />
low pitched voice, composed <strong>of</strong> a<br />
Figure 1. Audiospectrogram and waveform <strong>of</strong> a complete sequence <strong>of</strong><br />
four notes <strong>of</strong> <strong>the</strong> breeding call <strong>of</strong> Hyperolius kihangensis.<br />
0.2 0.4 0.6 0.9 1£ IjB<br />
S 0-0 02 0.4 Q& J.4 1j6
106 Biota 3/1-2,2002 AAENEGON & SALVIDIO<br />
Figure 2. Frequency spectrum and audiospectrogram <strong>of</strong> a single<br />
note <strong>of</strong> <strong>the</strong> breeding call <strong>of</strong> Hyperolius kihangensis.<br />
Table 1. Summary <strong>of</strong> numerical parameters <strong>of</strong> Hyperolius<br />
kihangensis advertisement call recorded at 19.7°C. kHz =<br />
kiloHertz, ms = milliseconds, S.D. = standard deviation.<br />
Call feature<br />
Fundamental frequency (kHz)<br />
Dominant frequency (kHz)<br />
Changes in dominant frequency<br />
(kHz)<br />
Note duration (ms)<br />
Internote duration (ms)<br />
sequence <strong>of</strong> four discrete notes. Every<br />
note had a slight variation in frequency<br />
(i.e., frequency modulation) and a peculiar<br />
tonal quality. The audiospectrogram<br />
depicts <strong>the</strong>se very characteristic features<br />
<strong>of</strong> <strong>the</strong> note (Figure 1). Two consecutive<br />
calls were recorded and analyzed; each is<br />
a sequence <strong>of</strong> four discrete notes, similar<br />
Mean ± S.D.<br />
2.076 ±0.1 96<br />
3.368 ±0.175<br />
782.5 ±113.7<br />
57 ± 6.23<br />
35 1.97 ±24.54<br />
Range<br />
2.225 - 3.208<br />
3.048 - 3.578<br />
666 - 956<br />
48.9 - 65.2<br />
321.9-381<br />
in fundamental and dominant frequency<br />
as well as note duration and frequency<br />
modulation. The intercall duration was<br />
about 20 seconds. As depicted in Figure<br />
2, <strong>the</strong> fundamental frequency <strong>of</strong> <strong>the</strong><br />
note was about 2.72 kHz and <strong>the</strong> dominant<br />
frequency 3.38 kHz. Each note had<br />
three to four peaks <strong>of</strong> intensity. The
MENEGON & SALVIDIO Biota 3/1-2,2002 107<br />
power spectrum and <strong>the</strong> audiospectrogram<br />
<strong>of</strong> a single note <strong>of</strong> <strong>the</strong> call were<br />
analyzed and are shown in Figure 2. The<br />
fundamental and dominant frequencies<br />
were highlighted and it is possible to<br />
appreciate that <strong>the</strong> note duration was<br />
about 56.5 ms. A summary <strong>of</strong> several call<br />
parameters is given in Table 1.<br />
DISCUSSION<br />
During our herpetological research in <strong>the</strong><br />
Udzungwa Scarp Forest Reserve, several<br />
mating pairs and egg clutches <strong>of</strong> H.<br />
kihangensis were observed. However,<br />
only one single male was heard calling<br />
and recorded. The very low frequency <strong>of</strong><br />
calling, combined with <strong>the</strong> brevity and<br />
quietness <strong>of</strong> <strong>the</strong> advertisement call <strong>of</strong> this<br />
species, could be reasons for <strong>the</strong> difficulty<br />
<strong>of</strong> recording vocalisations <strong>of</strong> H.<br />
kihangensis males. Thus, <strong>the</strong>re is a possibility<br />
that o<strong>the</strong>r Hyperolius species (H.<br />
tanneri and H. spinigularis) with welldeveloped<br />
gular sacs but with unknown<br />
voices (Schi0tz 1999, Schi0tz &<br />
Westergaard 2000) could emit short and<br />
very low-pitched calls, not easily recognizable.<br />
With respect to its breeding habitat and<br />
reproductive behaviour, H. kihangensis is<br />
quite different from all o<strong>the</strong>r Hyperolius<br />
species known for <strong>the</strong> Eastern Arc forests<br />
<strong>of</strong> Africa. Instead, it is comparable to<br />
some small, West African forest treefrogs<br />
such as H. zonatus and <strong>the</strong> larger H.<br />
bobirensis (Schi0tz 1999).<br />
The advertisement call <strong>of</strong> Hyperolius<br />
kihangensis is unusual within <strong>the</strong> genus,<br />
because <strong>of</strong> its peculiar tonal quality. A<br />
few species, such as H. kachalolae from<br />
Zambia and <strong>the</strong> West African H. viridigulosus,<br />
show similar features in <strong>the</strong>ir<br />
breeding calls. The calls <strong>of</strong> <strong>the</strong>se species<br />
are, however, clearly formed <strong>of</strong> a series <strong>of</strong><br />
harmonics. In <strong>the</strong> former, <strong>the</strong> fundamental<br />
and <strong>the</strong> dominant frequency are <strong>the</strong><br />
same, while in <strong>the</strong> latter <strong>the</strong> harmonics<br />
have a complex pattern <strong>of</strong> intensity maxima<br />
(Schi0tz .1999).<br />
Acknowledgements<br />
These data are part <strong>of</strong> a herpetological research project authorized by COSTECH<br />
(research permit # 98-028-CC-98-13). Special thanks are due to David Mover (Iringa)<br />
and Pr<strong>of</strong>. Kim Howell (University <strong>of</strong> Dar es Salaam) for helping us to obtain research<br />
permits and for many useful suggestions. We wish to thank Paolo Pedrini, Ivan<br />
Farronato and Martin Pickersgill for <strong>the</strong>ir comments on <strong>the</strong> preliminary version <strong>of</strong> <strong>the</strong><br />
manuscript<br />
REFERENCES<br />
CHARIF, R. A., MITCHELL, S. & CLARK, C. W. 1996: Canary 2.0 Users' manual. Cornell<br />
Laboratory <strong>of</strong> Ornithology, Ithaca, New York, USA.<br />
DUELLMAN, W.E. & TRUEB, L. 1986: Biology <strong>of</strong> Amphibians. Johns Hopkins University<br />
Press, Baltimore & London.<br />
MOREAU, R. E. 1935: A synecological study <strong>of</strong> Usambara, Tanganyka Territory, with particular<br />
reference to birds. Journal <strong>of</strong> Ecology 23: 1-43.<br />
SCHI0TZ, A. 1975: The Treefrogs <strong>of</strong> Eastern Africa. Steenstrupia, Copenhagen, Denmark.<br />
232 pp.<br />
SCHI0TZ, A. 1999: Treefrogs <strong>of</strong> Africa. Edition Chimaira, Frankfurt am Mein, Germany. 350<br />
PP-<br />
SCHI0TZ, A. & Westergaard, M. M. 2000: Notes on some Hyperolius (Anura: Hyperoliidae)
108 BJOla 3 1-2, 2002 MENEGON & SALVIDIO<br />
from Tanzania, with supplementary information on two recently described<br />
species. Steenstrupia 25: 1-9<br />
WESTEGAARD, M. M., BRESCIANI, J. & BUDTZ, P. E. 2000: Structural aspects <strong>of</strong> white spots<br />
on dorsal skin <strong>of</strong> Hyperolius kihangensis (Amphibia: Hyperoliidae). African<br />
Journal <strong>of</strong> Herpetology 4: 73-77.
NAGY JOGER, GUICKING & WINK 3/1-2, 2OO2 109<br />
Phylogeography <strong>of</strong> <strong>the</strong> European<br />
Whip Snake Coluber (Hierophis)<br />
viridiflavus as inferred from<br />
nucleotide sequences <strong>of</strong> <strong>the</strong><br />
mitochondrial cytochrome b gene<br />
and ISSR genomic fingerprinting<br />
Zoltan Tamas NAGY1, Ulrich JOGER2, Daniela<br />
GUICKING1 & Michael WINK1<br />
1lnstitut fur Pharmazie und Molekulare Biotehnolgie, Universitat Heidelberg, Im<br />
Neuenheimer Feld 364, D-69120 Heidelberg, Germany<br />
2Hessisches Landesmuseum Darmstadt, Zoologische Abteilung, Friedensplatz 1, D-<br />
64283 Darmstadt, Germany<br />
Abstract<br />
The intraspecific phylogeography <strong>of</strong> European Whip Snake Coluber (Hierophis) viridiflavus<br />
was reconstructed using complete sequences (1117 bp) <strong>of</strong> <strong>the</strong> protein-coding<br />
mitochondrial cytochrome b gene. C. (H.) gemonensis (Laurenti) and C. (H.) caspius<br />
Gmelin were used as outgroups. Additionally, a microsatellite-based genomic fingerprint<br />
method, ISSR, was employed to check for gene flow between populations. Two clearly<br />
different genetic clades could be identified within European Whip Snake, a western one<br />
occurring in France, Switzerland and Italy west <strong>of</strong> <strong>the</strong> Apennines, and an eastern one<br />
found in Croatia, eastern and sou<strong>the</strong>rn Italy. The latter one could be fur<strong>the</strong>r subdivided<br />
into three subgroups, two <strong>of</strong> which occur in sou<strong>the</strong>rnmost Italy only (sou<strong>the</strong>rn Calabria<br />
and Sicily). These two distinct entities were probably formed during a long continuous<br />
settlement <strong>of</strong> <strong>the</strong> European Whip Snake in <strong>the</strong>se climatically favourable areas, whereas<br />
<strong>the</strong> more nor<strong>the</strong>rn populations experienced enormous shrinking <strong>of</strong> <strong>the</strong>ir ranges during<br />
cold periods in <strong>the</strong> Pleistocene. Potential glacial refuge areas are discussed.<br />
Key words: European Whip Snake, Coluber (Hierophis) viridiflavus, phylogeography<br />
Received 16 November 2001; accepted 5 February 2002
110 >ta 3/1-2, 2002 NAGY ,JOGER, GUICKING & WINK<br />
INTRODUCTION<br />
The taxonomy and systematics <strong>of</strong> <strong>the</strong><br />
European Whip Snake, Coluber<br />
(Hierophis) viridiflavus Lacepede, 1789,<br />
and <strong>of</strong> <strong>the</strong> o<strong>the</strong>r species <strong>of</strong> <strong>the</strong> genus<br />
Coluber (sensu lato), have been intensively<br />
debated in <strong>the</strong> last centuries.<br />
Although <strong>the</strong> European whip snake exists<br />
exclusively in Europe (Heimes 1993,<br />
Naulleau 1997), it shows remarkable<br />
morphological variation. Many <strong>of</strong> <strong>the</strong>se<br />
characteristics can change from population<br />
to population (Schatti & Vanni<br />
1986), and <strong>the</strong>refore a multitude <strong>of</strong> morphological<br />
forms (insular and colour<br />
forms first <strong>of</strong> all) at species and subspecies<br />
levels have been reported in <strong>the</strong><br />
literature (Suckow 1798, Bonaparte<br />
1833, De Betta 1874, Boulenger 1893,<br />
Mertens & Muller 1928, Mertens &<br />
Wermuth 1960, Kramer 1971,<br />
Capolongo 1984, etc.). While Bruno<br />
(1975 & 1980) considers that two separate<br />
taxa ("viridiflavus" and "carbonarius"<br />
- on <strong>the</strong> basis <strong>of</strong> melanism) at species<br />
level might be possible, Schatti & Vanni<br />
(1986) among o<strong>the</strong>rs question <strong>the</strong> existence<br />
<strong>of</strong> subspecies - due to lack <strong>of</strong> evidence<br />
in pholidotic and hemipenis-morphological<br />
data. The occurence <strong>of</strong> <strong>the</strong><br />
Table 1. Tissue samples included in <strong>the</strong> present study.<br />
* - no proper data available<br />
Sample Species Country <strong>of</strong> origin<br />
P7 (1) C. (Hjviridifloms Croatia<br />
J19(5) " France<br />
J36(3) " Italy<br />
J39(5) " France<br />
MO (5)<br />
DOS (7)<br />
DG9(7)<br />
DG16(9)<br />
DG17
NAGY JOGER, GUICKING & WINK Biota 3/i-a, 2002 111<br />
Figure 1. Map with localities <strong>of</strong> specimens used for <strong>the</strong> analysis.<br />
Switzerland, see Table 1 and Figure 1 for<br />
details). In most cases a blood sample<br />
was taken from <strong>the</strong> tail <strong>of</strong> living specimens<br />
at <strong>the</strong> capture site (Joger & Lenk<br />
1997); in addition, different tissue samples<br />
were taken (a small piece <strong>of</strong> liver,<br />
heart, or tail) from recently killed animals<br />
(snakes that were run over by cars). The<br />
distribution area <strong>of</strong> <strong>the</strong> European whip<br />
snake (Heimes 1993, Naulleau 1997) was<br />
more or less covered by sampling - with<br />
<strong>the</strong> exception <strong>of</strong> <strong>the</strong> Western islands <strong>of</strong><br />
<strong>the</strong> Mediterranean Sea, such as Corsica,<br />
Sardinia etc. Samples from <strong>the</strong> two closely<br />
related species (Schatti 1988): Coluber<br />
(Hierophis) gemonensis (Laurenti, 1768)<br />
and Coluber (Hierophis) caspius (Gmelin,<br />
1789), were analyzed as outgroups.<br />
2. Isolation <strong>of</strong> DMA<br />
Tissue samples were stored until processing<br />
ei<strong>the</strong>r in 80% ethanol or in EDTA<br />
buffer (Arctander 1988). The isolation <strong>of</strong><br />
total genomic DMA was carried out with<br />
proteinase K digestion, followed by<br />
cleaning steps with phenol-chlor<strong>of</strong>orm<br />
and guanidine-thiocyanate (Sambrook et<br />
al. 1989). The extracted DNA was dissolved<br />
and stored in Tris-EDTA (10 mM<br />
Tris and 1mM EDTA).<br />
3. PCR and nucleotide sequencing<br />
The whole cytb gene was amplified with<br />
polymerase chain reaction in an end volume<br />
<strong>of</strong> 50 ul. PCR primers are listed in<br />
Table 2. PCR was carried out under standard<br />
conditions: initial denaturating (4<br />
min at 94°C), 31-33 cycles with denaturating<br />
(45 s, 94°C), elongation (2 min,<br />
72°C) and annealing step (50 s, 41-<br />
45°C), followed by a final elongation step<br />
for 10 min at 72°C. PCR products were<br />
stored at 4°C, and were directly used as<br />
templates in a cycle sequencing reaction<br />
(Lenk et al. 1999). Typical conditions<br />
were: 3 min at 94°C, 26 cycles with<br />
denaturating (30 s, 94°C) and annealing<br />
and elongation step (45 s, 60°C). The<br />
sequencing products were analyzed in an<br />
automated sequencer ALFExpress II<br />
(Amersham Pharmacia Biotech).<br />
Sequences were checked for errors and
112 oiola 3/1-2,2002 NAGY rJOGERr GUICKING & WINK<br />
Table 2. Primers used for PCR amplification and sequencing (cy: primer used in cycle<br />
sequencing only).<br />
Primer<br />
L14724<br />
smiA (L-14S46)<br />
mtE (H-15556)<br />
smtF (H-16060)<br />
L-15570cy<br />
H-15305cy<br />
Source<br />
Meyer eial. (1990)<br />
after Kocher et al. ( 1989),<br />
Leak &Wink (1997)<br />
Lenketal. (2001)<br />
modified after Wink (1995)<br />
This study<br />
This study<br />
aligned manually.<br />
The statistical analyses <strong>of</strong> <strong>the</strong> nucleotide<br />
sequences were carried out with <strong>the</strong><br />
PAUP* 4.0b8 (Sw<strong>of</strong>ford 2001) programme<br />
package using maximum parsimony<br />
method with heuristic search and<br />
bootstrap analysis.<br />
4. ISSR-PCR fingerprinting<br />
ISSR (Inter simple sequence repeat)<br />
genomic fingerprinting was performed to<br />
verify results obtained from mitochondrial<br />
cytochrome b sequences and to evaluate<br />
potential hybridisation. ISSR-PCR<br />
uses single primers designed from short<br />
tandem repeats to amplify stretches <strong>of</strong><br />
DNA between adjacent microsatellites.<br />
Given that microsatellites are scattered<br />
throughout <strong>the</strong> genome (Tautz & Renz<br />
1984) and in such density that adjacent<br />
microsatellites lie within <strong>the</strong> limits <strong>of</strong> Taq<br />
polymerase processing, during ISSR-PCR<br />
a large number <strong>of</strong> fragments is generated.<br />
Once separated on polyacrylamide<br />
gel, ISSR-PCR products appear as polymorphic<br />
multiband fingerprints.<br />
Five ISSR-primers that are known to generate<br />
polymorphic fingerprints in several<br />
vertebrate taxa were tested under varying<br />
PCR conditions on Coluber<br />
(Hierophis) samples. (CA)io- and<br />
(GACA)4-primers gave most useful<br />
results. The reactions were carried out<br />
according to <strong>the</strong> protocol <strong>of</strong> Wink et al.<br />
Sequence (S'-3')<br />
CGA AGC TTG ATA TGA AAA ACC ATC<br />
CAA CAT CTC AGC ATG ATG AAA CTT CG<br />
AAT AGG AAG TAT CAT TCT GGT TTG AT<br />
TCA GTT TTTGGT TTA CAA GAC CAA TG<br />
GAY AAA ATC CCA TTY CAC CC<br />
AAT GAT ATT TGT CCT CAT GG<br />
(2000). In a PCR volume <strong>of</strong> 25 ul, 50-100<br />
ng <strong>of</strong> total DNA were used as template,<br />
plus 6 pmol (GACA)4 or 20 pmol (CA)io<br />
primer, 1,5 mM MgCb, 0,1 mM <strong>of</strong> dCTP,<br />
dGTP, and dTTP, 0,045 mM dATP, 1 uCi<br />
33P-alpha-dATP, 2,5 ul 10x amplification<br />
buffer and 1 unit Taq polymerase<br />
(Amersham Pharmacia Biotech). PCR<br />
programmes were set for 5 min at 94°C,<br />
followed by 26 cycles <strong>of</strong> 30 s at 94°C, 20<br />
s at annealing-temperature, and 50 s at<br />
72°C. After completion, <strong>the</strong> temperature<br />
was set to 72°C for 5 min and <strong>the</strong>n lowered<br />
to 4°C for fur<strong>the</strong>r storage. Optimal<br />
annealing-temperatures, as found out by<br />
a 2°C temperature gradient PCR (40°C-<br />
60°C), were 40°C for (CA)io-primer and<br />
55°C for (GACA)4-primer. PCR products<br />
were separated on a denaturing Sequagel<br />
matrix at 65 W for 4 h and visualised by<br />
autoradiography. Fingerprint patterns<br />
were evaluated visually.<br />
RESULTS<br />
1. Analysis <strong>of</strong> cytochrome b sequences<br />
The complete mitochondrial gene cytb<br />
(1117 base pairs) was amplified from 24<br />
samples, including <strong>the</strong> outgroups.<br />
The phylogeographic analysis <strong>of</strong> Coluber<br />
(Hierophis) viridiflavus revealed four haplotype<br />
groups in two main clades, which<br />
are clearly separated genetically and by<br />
geographic areas (Figure 2). These clades<br />
are supported with significant bootstrap
NAGY ,JOGERr GUICKING & WINK 3/1-2, 2002 113<br />
values (96-100%).<br />
Among our sequences 57 variable<br />
nucleotides (5.10% <strong>of</strong> <strong>the</strong> cytb gene,<br />
excluding outgroups) were found (Figure<br />
3); <strong>the</strong> two main groups are separated by<br />
- at least - 2.95% (33 different<br />
nucleotides). They represent a Western<br />
and an Eastern clade. The Western group<br />
(clade W in Figure 2) includes animals<br />
from France, Switzerland and Italy west<br />
<strong>of</strong> <strong>the</strong> Apennines (i.e. Tuscany province<br />
and Riviera coast). This group possesses a<br />
quite uniform haplotype - only single<br />
nucleotide mutation sites were detected -<br />
which appears to be monomorphic without<br />
fur<strong>the</strong>r subdivisions.<br />
The Eastern group is divided into fur<strong>the</strong>r<br />
subgroups: 1. Italy, east <strong>of</strong> <strong>the</strong> Apennines<br />
and <strong>the</strong> island Krk (clade E on Figure 2),<br />
2. sou<strong>the</strong>rn Calabria, and 3. Sicily (both<br />
<strong>of</strong> <strong>the</strong>m represented in clade S). At <strong>the</strong><br />
Eastern border <strong>of</strong> <strong>the</strong> distribution area live<br />
- sympatrically with C. (H.) gemonensis -<br />
<strong>the</strong> Slovenian and Croatian populations<br />
<strong>of</strong> C. (H.) viridiflavus. Our sample from<br />
<strong>the</strong> island Krk shows <strong>the</strong> closest relationship<br />
with <strong>the</strong> eastern Italian samples. In<br />
contrast, <strong>the</strong> Sou<strong>the</strong>rn (South-Eastern)<br />
populations belong to a clearly separated<br />
group: <strong>the</strong> South Calabrian and Sicilian<br />
populations differ not only from <strong>the</strong><br />
nor<strong>the</strong>astern ones, but also from each<br />
o<strong>the</strong>r.<br />
2. ISSR-PCR patterns<br />
ISSR-primers (GACA)4 and (CA)io produced<br />
informative fingerprints with several<br />
polymorphic bands. ISSR pr<strong>of</strong>iles<br />
were more diverse with <strong>the</strong> (GACA)4primer<br />
(Figure 4) than with <strong>the</strong> (CA)ioprimer<br />
(not shown). Both primers generate<br />
a series <strong>of</strong> species-specific bands as<br />
Figure 2. Phylogeny <strong>of</strong> <strong>the</strong> cytochrome b gene <strong>of</strong> Coluber (Hierophis) viridiflavus. Strict<br />
consensus tree resulted from maximum parsimony analysis, tree length=251. Bootstrap<br />
proportions >50 (100 replications) are presented.<br />
100<br />
cyt6-MP<br />
100<br />
W<br />
99<br />
Krk, CRO (P7)<br />
Canossa, I (155}<br />
F. Staffora, I (DG77)<br />
T. Platano, I (DG66)<br />
T. Platano, I (DG65)<br />
N-Calabria, I (DG5S}<br />
Torre S. Genaro, 1 (DG22)<br />
Mte St. Angelo, I (DG21)<br />
Lago di Lesina, I (DG17}<br />
Lago di Lesina, 1 (DG16)<br />
Grado, \(t>G9)<br />
Grade, I (DCS;<br />
9gj- Sicily, I (DG38)<br />
63 PL Sicily. I (DG45)<br />
I S-Calabria, S-Cala I (DG53)<br />
Quincy, F (J19)<br />
Canguers, F (Al)<br />
R. L'AIlondon, CH (DG86)<br />
Rapallo, I (DG7I)<br />
Quincy, F (J40)<br />
Quincy, F (J39)<br />
Tuscany, I (J36)<br />
C. (H.) gemonensis, CRO<br />
C. (H.) cospius, OR
114 Biota 3 1-2. 3002 NAGY ,JOCER, GUICKING & WINK<br />
Figure 3. Variable nucleotides <strong>of</strong> C. (H.) viridiflavus samples (cytochrome b gene with<br />
position numbering). E: eastern group; W: western group; Sic: Sicily; Cal: South-<br />
Calabria; var: variable sequences with single nucleotide mutations.<br />
*c<br />
*c<br />
#C<br />
*c_<br />
SC<br />
K<br />
vi<br />
vi<br />
vi<br />
vi<br />
vi<br />
vi<br />
id<br />
id<br />
id<br />
id<br />
id<br />
id<br />
1111<br />
111 1111222233 3344444445 5555666666 7778888999 9990000<br />
2244458122 4569056811 3834566671 3457146889 0261447011 5790467<br />
4527814709 7658487828 0127302577 1929829341 8934366658 5492721<br />
E<br />
E var<br />
E~Sic<br />
E Cal<br />
W<br />
W<br />
var<br />
GCACACGCGA TTTGGCTCGG<br />
. . .G.TRT AT.T . .AC<br />
. . .G. . .TA- .C...A.C. . .AC<br />
ATGGG.A. .G<br />
ATGGG.A.<br />
TGTTATAGTC<br />
R<br />
GGACCAATTC<br />
Y W<br />
T.G.<br />
G.C<br />
ACGAGGCAAT ATGAGTC<br />
D<br />
Y<br />
. . . GA. . . . . AG . . .<br />
. G.A .C.<br />
. .CAA.C.AA CA.CGA.AC. AA.T CT GT . .AA.G.C ... .A.T<br />
G Y. CAA.C.AA CA.CGA.AC . AA.T. . CT . . GT. .AAYGMC . . . .A.T<br />
... . irrn n/-n ,. . , ...<br />
Figure 4. ISSR PCR fingerprint made with<br />
, ° ,-.,„ . .. , , , , . .,<br />
(GACA)4 primer. See text for details.<br />
'<br />
well as several group-specific bands<br />
.... . ,. ° £, K , . ,<br />
(Figure 4. indicated with numbers) that<br />
s . ,' ,.„<br />
support <strong>the</strong> differentiation into a western<br />
(Figure 4, bands No. 1, 2, 3, 4) and an<br />
eastern clade (bands No. 5: several bands<br />
in a specific size range, to a certain extent<br />
also No. 6, 7). Western and eastern<br />
clades, as suggested by ISSR-analysis,<br />
conform to those suggested by<br />
cytochrome b sequences.<br />
Fur<strong>the</strong>rmore, ISSR patterns suggest a<br />
\ f--•- * hybrid status for some samples. This is<br />
evident in samples J55 and DG77, both<br />
from Northwestern Italy. According to<br />
i cytochrome b sequences, both samples<br />
7 belong to <strong>the</strong> eastern clade, whereas<br />
_ _. ,. _^__ „ ISSR-pr<strong>of</strong>iles show both eastern clade<br />
"* specific bands (Figure 4, no. 5) as well as<br />
• "" rzim;^_ 6 western clade specific bands (in sample<br />
•^^V^"-—-^35— 4 DG77 No 2 and 3^ in samp|e J55 No 1;<br />
^"^•"•-^--"""^ '-. ' 2, and 3). These results suggest recent<br />
gene flow between <strong>the</strong> eastern and western<br />
clade in Northwestern Italy.<br />
The separation <strong>of</strong> <strong>the</strong> Sicilian and sou<strong>the</strong>rn<br />
Caiabrian samples (DG38, DG45 and<br />
DG53, respectively) from <strong>the</strong> rest <strong>of</strong> <strong>the</strong><br />
eastern clade is less evident according to<br />
*•==»* ~ ISSR pr<strong>of</strong>iles than it is according to<br />
cytochrome b sequences. The band pattern<br />
<strong>of</strong> <strong>the</strong> Sicilian sample DG45 may<br />
show genetic introgression from <strong>the</strong><br />
west, too (Figure 4, bands No. 2, 3).<br />
DISCUSSION<br />
The sequence analysis (confirmed by
NAGY ,JOGER, GUICKING & WINK Biota 3/i-a, 2002 115<br />
ISSR-PCR fingerprinting) unambiguously<br />
shows that <strong>the</strong>re are discrete phylogeographic<br />
units within <strong>the</strong> distribution range<br />
<strong>of</strong> <strong>the</strong> European Whip Snake. Spatial and<br />
historic factors might be responsible for<br />
<strong>the</strong>ir isolation.<br />
Two bigger groups are apparently separated<br />
on <strong>the</strong> genetic level: The Western<br />
and Eastern populations are divided by<br />
<strong>the</strong> mountain ranges <strong>of</strong> <strong>the</strong> Alps and <strong>the</strong><br />
Apennines acting as strict geographic<br />
barriers (Figure 1). Apparently this species<br />
does not cross mountain ranges easily<br />
(though it has been reported from elevations<br />
above 1,800 m [Heimes 1993]).<br />
Results based on cytb sequences support<br />
<strong>the</strong> conclusion that <strong>the</strong> Western group<br />
might be monotypic; however, no samples<br />
from <strong>the</strong> Western Mediterranean<br />
islands (see above), or from <strong>the</strong> Nor<strong>the</strong>rn<br />
border <strong>of</strong> <strong>the</strong> distribution zone (e.g.<br />
Luxembourg) were available for our<br />
research. An important aspect is that <strong>the</strong><br />
distribution <strong>of</strong> this colubrid species is continuous<br />
between France and <strong>the</strong> Western<br />
foot <strong>of</strong> <strong>the</strong> Apennines; it includes a recent<br />
connection along <strong>the</strong> narrow coastal strip<br />
<strong>of</strong> Cote d'Azur and Riviera di Ponente.<br />
The mitochondria! sequences confirm a<br />
close relationship between Coluber<br />
(Hierophis) gemonensis and Coluber<br />
(Hierophis) viridiflavus. This result suggests<br />
also a Western-Eastern divergence:<br />
Coluber (Hierophis) viridiflavus is representative<br />
<strong>of</strong> <strong>the</strong> central Mediterranean,<br />
while Coluber (Hierophis) gemonensis <strong>of</strong><br />
<strong>the</strong> eastern Mediterranean, respectively.<br />
In addition, it is also possible that both <strong>of</strong><br />
<strong>the</strong>m share a common ancestor with C.<br />
(H.) caspius and C. (H.) jugularis, as<br />
Schatti (1988) believes.<br />
As <strong>the</strong> sister species are distributed in <strong>the</strong><br />
Balkans, it is justified to assume <strong>the</strong> geographic<br />
origin <strong>of</strong> C. (H.) viridiflavus at <strong>the</strong><br />
eastern edge <strong>of</strong> its distribution range.<br />
Many eastern European species that<br />
reach Italy do not cross <strong>the</strong> Apennines; a<br />
vertebrate example is <strong>the</strong> Hooded Crow<br />
Corvus corone comix. In <strong>the</strong> turtle Emys<br />
orbicularis, genetically different subspecies<br />
inhabit eastern and western Italy<br />
(Lenketal. 1999).<br />
The population living on Krk island - at<br />
<strong>the</strong> eastern border <strong>of</strong> <strong>the</strong> distribution<br />
zone - shows a petty divergence compared<br />
to Italian populations which belong<br />
to <strong>the</strong> eastern group. A continuous presence<br />
<strong>of</strong> <strong>the</strong> species during <strong>the</strong> Pleistocene<br />
is, however, unlikely in this nor<strong>the</strong>rn part<br />
<strong>of</strong> its range. Therefore it is probably<br />
derived from a sou<strong>the</strong>rn Italian stock (The<br />
Balkans' own glacial refugia were probably<br />
occupied by C. (H.) gemonensis).<br />
North-South movements <strong>of</strong> such reptiles<br />
along <strong>the</strong> coasts <strong>of</strong> Italy are unhindered.<br />
The existence <strong>of</strong> South(east)ern subgroup^)<br />
is, however, again paralleled by<br />
Emys orbicularis (Lenk et al. 1999) as well<br />
as by <strong>the</strong> snakes <strong>of</strong> <strong>the</strong> Elaphe longissima<br />
complex (Lenk & Wuster 1999). Such<br />
common zoogeographical patterns can<br />
be traced back to <strong>the</strong> glacial zoogeographic<br />
history in all likelihood (Lenk et<br />
al. 1999, Schatti 1988).<br />
The sou<strong>the</strong>rn Italian and Sicilian regions<br />
could have functioned as refugia during<br />
cold periods <strong>of</strong> <strong>the</strong> Pleistocene. A refuge<br />
for <strong>the</strong> eastern group could have been at<br />
<strong>the</strong> Gulf <strong>of</strong> Taranto and in Calabria. Our<br />
two Calabrian samples belong to slightly<br />
different haplotypes: DG53 ("South"),<br />
and DG55 ("North"), respectively. Both<br />
are different from <strong>the</strong> Sicilian sample. This<br />
variation could be explained by a longer<br />
continuous evolutionary history in <strong>the</strong>se<br />
possible glacial refuges. A possible refuge<br />
for <strong>the</strong> western group could have been in<br />
<strong>the</strong> region <strong>of</strong> Naples-Salerno. However,<br />
we could not check samples from that<br />
area.<br />
The evolution <strong>of</strong> <strong>the</strong> sou<strong>the</strong>rn populations<br />
could also continue unhindered during<br />
glacial periods. This may have resulted<br />
in higher genetic differences, in o<strong>the</strong>r<br />
words, in a mosaic <strong>of</strong> genetically different<br />
forms. A few <strong>of</strong> <strong>the</strong>se forms (in fact only
116 Biota 3/i-a, 2002 NACY ,JOGER, GUICKING & WINK<br />
one on each side <strong>of</strong> <strong>the</strong> Apennines)<br />
would have used <strong>the</strong> climatically favorable<br />
interglacials to spread into more<br />
nor<strong>the</strong>rn areas. Pleistocene fossils attributed<br />
to C. (H.) viridiflavus have been<br />
found in sou<strong>the</strong>rn Germany and Austria;<br />
in <strong>the</strong> Pliocene, this species reached <strong>the</strong><br />
territories <strong>of</strong> today's Czech Republic and<br />
Poland (Ivanov 1997, Schatti 1988).<br />
Climatic conditions during <strong>the</strong> glacial<br />
periods forced it to retreat into <strong>the</strong> above<br />
named refuges in sou<strong>the</strong>rn Italy. After <strong>the</strong><br />
last glacial period, re-immigration to <strong>the</strong><br />
nor<strong>the</strong>rn Adriatic region (eastern group)<br />
as well as to France and Belgium (western<br />
group) would have happened in a comparatively<br />
short time, which may not<br />
have allowed a significant genetic differentiation<br />
<strong>of</strong> nor<strong>the</strong>rn populations to<br />
occur.<br />
ISSR-PCR has useful applications for<br />
examining group specificity and<br />
hybridization events. Because <strong>of</strong> its<br />
detection <strong>of</strong> nuclear genetic rearrangements,<br />
it provides a valuable complement<br />
to data obtained from <strong>the</strong> maternally<br />
inherited mitochondrial genome. It is<br />
especially powerful in detecting<br />
hybridization events (Wink et al. 2000).<br />
In our study, it allowed us to re-examine<br />
<strong>the</strong> separation <strong>of</strong> Coluber (Hierophis)<br />
viridiflavus samples into a western and an<br />
eastern clade, which is strongly supported<br />
by ISSR-analysis, as well as suggesting a<br />
gene flow between both groups in<br />
Northwestern Italy. However, ISSR-fingerprints<br />
provided less evidence for <strong>the</strong><br />
separation <strong>of</strong> Sicilian and South Calabrian<br />
samples from o<strong>the</strong>r eastern clade samples<br />
than is suggested by cytochrome b<br />
sequence data.<br />
Because <strong>of</strong> <strong>the</strong> evident, but not complete,<br />
genetic isolation, we recommend once<br />
more <strong>the</strong> recognition <strong>of</strong> two subspecies<br />
<strong>of</strong> <strong>the</strong> European whip snake sensu<br />
Mertens & Wermuth (1960). If fur<strong>the</strong>r<br />
studies in <strong>the</strong> contact zones <strong>of</strong> both subspecies<br />
prove that an intrinsic genetic<br />
barrier exists, <strong>the</strong> two main clades could<br />
even be raised to species level (see Joger<br />
et al. 1998). The applicable name for <strong>the</strong><br />
western populations is Coluber<br />
(Hierophis) viridiflavus viridiflavus<br />
Lacepede, 1789 (terra typica: sou<strong>the</strong>rn<br />
France). The Eastern stocks were traditionally<br />
assigned to <strong>the</strong> subspecies<br />
Coluber (Hierophis) viridiflavus carbonarius<br />
Bonaparte 1833 (nomen conservandum,<br />
terra typica restricta Mertens &<br />
Muller 1928: Monti Euganei, Padua,<br />
Italy); however, <strong>the</strong> subspecific name has<br />
no more (or has far less) ethymological<br />
importance (an allusion to <strong>the</strong> melanism).<br />
The border between both subspecies is<br />
<strong>the</strong> Apennines. The South Italian populations<br />
are less differentiated within <strong>the</strong><br />
Eastern group (1.16% [13 different<br />
nucleotides] and 1.34% [15] by<br />
Calabrian and Sicilian samples, respectively,<br />
at cytb level), so in our view taxonomical<br />
consequences are not justified.<br />
This, however, would mean that older<br />
names apply to <strong>the</strong> eastern subspecies:<br />
Coluber (Hierophis) pr<strong>of</strong>ulax Costa, 1828<br />
(terra typica Aspromonte, sou<strong>the</strong>rn<br />
Calabria) or C. (H.) xanthurus<br />
Rafinesque-Schmaltz, 1810 (terra typica<br />
Sicily). An even older name is C. (H.) sardus<br />
Suckow 1798 (Terra typica Sardinia).<br />
As we have not studied Sardinian material,<br />
we cannot decide to which form it<br />
applies. In all cases, <strong>the</strong> morphology <strong>of</strong><br />
<strong>the</strong> European whip snake has to be studied<br />
in more detail in order to find reliable<br />
characters to define subspecies.<br />
Acknowledgements<br />
We would like to thank Toni Amann, Anna Hundsdorfer, Luc Legal, Peter Lenk and<br />
Werner Mayer for providing tissue samples, as well as Edoardo Razzetti for helpful comments.<br />
This study was financially funded by DAAD (research scholarship for <strong>the</strong> first<br />
author) and DFG (Jo-134-7 and Wi-719/18).
NAGY JOGER, GUICKING & WINK Biota 3/1-2, 2002 117<br />
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1<br />
PASTOREILI, LAGHI & SCARAVELLI Biota 3/1-2. 2002 119<br />
Seasonal activity and spatial<br />
distribution <strong>of</strong> a Speleomantes<br />
italicus population in a natural<br />
cave<br />
Christian PASTORELLI1, Paolo LAGHI2 & Dino<br />
SCARAVELLI3<br />
Y Cerchia di S. Egidio 2205, 47023 Cesena (FC), Italy<br />
E-mail: pastorellic@libero.it<br />
2v. Bruno C. Garibaldi 22, 47100 Forli, Italy<br />
E-mail: spelerpes@libero.it<br />
3Riserva Naturale Orientata e Museo di Onferno, p. Roma 1, 47855 Gemmano (RN),<br />
Italy<br />
E-mail: rnoonf@tin.it<br />
Abstract<br />
The biology <strong>of</strong> a Speleomantes italicus population inhabiting <strong>the</strong> natural cave "Grotta<br />
del Tritone" has been studied since 1998. The cave opens at 810 m a.s.l. and it is located<br />
in <strong>the</strong> nor<strong>the</strong>rn Apennine slope (Forli-Cesena province, Emilia-Romagna region,<br />
Italy). Data on environmental parameters (inside and outside temperature, relative<br />
humidity and light intensity) and activity <strong>of</strong> Speleomantes italicus (number, size and<br />
location <strong>of</strong> <strong>the</strong> individuals) have been collected from February 1999 to January 2001 by<br />
monthly surveys. The hypogean observable activity <strong>of</strong> 5. italicus in <strong>the</strong> study site<br />
showed a strong seasonal pattern, with <strong>the</strong> highest number <strong>of</strong> specimens in late spring<br />
(May) and early fall (September). The authors also analysed seasonal variation <strong>of</strong> spatial<br />
distribution as a function <strong>of</strong> temperature, humidity and light intensity. Salamanders<br />
were observed only in <strong>the</strong> first part <strong>of</strong> <strong>the</strong> cave (up to about 25 meters from <strong>the</strong><br />
entrance); <strong>the</strong> animals also seemed to prefer <strong>the</strong> lighted parts <strong>of</strong> <strong>the</strong> cave, while juveniles<br />
were found at a shorter distance from <strong>the</strong> entrance than adults. During summer,<br />
salamanders were active deeper in <strong>the</strong> cavity than in fall or spring, and in winter <strong>the</strong>y<br />
were found near <strong>the</strong> entrance and sometimes outside. Superficial activity was scarce<br />
and close to cave entrance.<br />
Key words: population ecology, seasonal activity, spatial distribution, Plethodontidae,<br />
Speleomantes italicus<br />
Received 3 August 2002; accepted 26 September 2002
120 Biota 3/i-a, 2002 PASTORELLI, LAGHI & SCARAVELLI<br />
INTRODUCTION<br />
European Plethodontid salamanders <strong>of</strong><br />
<strong>the</strong> genus Speleomantes are <strong>full</strong>y terrestrial<br />
and live in moist and cool environments<br />
such as caves and soil interstices.<br />
They are found from south-eastern<br />
France to central Italy and on <strong>the</strong> island<br />
<strong>of</strong> Sardinia (Lanza et al. 1995). To date,<br />
seven species (three continental and four<br />
insular) and one insular subspecies <strong>of</strong><br />
Speleomantes have been recognised on<br />
<strong>the</strong> basis <strong>of</strong> morphological and genetic<br />
data (Lanza et al. 1986, 1995, 2001,<br />
Nascetti et al. 1996). Notwithstanding<br />
<strong>the</strong> great interest that this genus holds,<br />
studies on eco-ethology, population<br />
dynamics and life history <strong>of</strong><br />
Speleomantes started only recently and<br />
regarded few species (Cimmaruta et al.<br />
1999, Mutz 1998, Pastorelli et al. 2001,<br />
Salvidio 1990, 1991, 1992, 1993a,<br />
1993b, 1996, 1998; Salvidio et al. 1994,<br />
Voesenek et al. 1987).<br />
Speleomantes italicus is a species endemic<br />
to <strong>the</strong> nor<strong>the</strong>rn and central Apennines<br />
(Lanza et al. 1995) and is found in natural<br />
and artificial caves and in interstitial<br />
habitats.<br />
The biology <strong>of</strong> a Speleomantes italicus<br />
population inhabiting a natural cave in<br />
<strong>the</strong> nor<strong>the</strong>rn Apennines has been studied<br />
since 1998 (Pastorelli et al. 2001). This<br />
paper reports data about seasonal activity<br />
and spatial distribution <strong>of</strong> <strong>the</strong> salamanders<br />
as a function <strong>of</strong> environmental<br />
parameters studied from February 1999<br />
to January 2001.<br />
MATERIALS AND METHODS<br />
The study site is a natural cave called<br />
"Grotta del Tritone" [43°53'52" N -<br />
0°29'02"W (Rome)] located on <strong>the</strong><br />
nor<strong>the</strong>rn Apennine slope (Forli-Cesena<br />
province, Emilia-Romagna Region, Italy).<br />
The cave opens north-eastward at 810 m.<br />
a.s.l., in marl and sandstones. It reaches a<br />
total extension <strong>of</strong> about 50 meters, on a<br />
sub-horizontal horizon. (Figure 1). The<br />
dominant vegetation around <strong>the</strong> study<br />
site is oak-tree woodland, mainly composed<br />
<strong>of</strong> coppices <strong>of</strong> Quercus cerris and<br />
Ostrya carpinifolia with a poor leaf soil.<br />
Since September 1998 <strong>the</strong> study site was<br />
sampled monthly (except June 1999) and<br />
records included individual data as well as<br />
environmental data.<br />
Salamanders were searched for by two<br />
persons outside <strong>the</strong> cave near its<br />
Figure 1. The cave "Grotta del Tritone". On <strong>the</strong> left <strong>the</strong> black circle indicates <strong>the</strong> location<br />
<strong>of</strong> <strong>the</strong> study site in Italy and in grey is pointed out Speleomantes italicus range.<br />
GROTTA DHL TRiTONE
PASTORELLI, LAGHI & SCARAVELLI Biota 3 1-2, 2OO2 121<br />
entrance, and inside on <strong>the</strong> walls and<br />
floor and, where possible, on <strong>the</strong> cave<br />
vault for about one hour per session. For<br />
each specimen captured <strong>the</strong> following<br />
data were recorded: date and hour <strong>of</strong><br />
observation, sex, distance from cave<br />
entrance and from cave floor (in meters).<br />
All specimens with a body length wider<br />
than <strong>the</strong> smallest male (41 mm) with a<br />
mental body gland (cf. Lanza 1959) were<br />
considered to be adult salamanders.<br />
Environmental data include wea<strong>the</strong>r conditions,<br />
temperatures and relative humidity.<br />
The latter two were measured at <strong>the</strong><br />
entrance (Te1, URe1) and at <strong>the</strong> following<br />
distances from it: 5 meters outside <strong>the</strong><br />
cave (Te2, URe2) and 7 (Ti1, URi1), 15<br />
012, URi2) and 21 metres (Ti3, URi3)<br />
inside. Two minimum-maximum mercury<br />
<strong>the</strong>rmometers were placed to record temperatures<br />
between consecutive samples,<br />
respectively at Te2 and Ti1 sampling locations.<br />
Finally, in July 1999, March, June<br />
and December 2000 maximum light<br />
intensity was recorded for each linear<br />
meter inside <strong>the</strong> cave to <strong>the</strong> point at<br />
which light reached <strong>the</strong> 0.0 lux instrumental<br />
value.<br />
RESULTS<br />
Environmental parameters<br />
During <strong>the</strong> study period, <strong>the</strong> mean minimum<br />
temperature outside <strong>the</strong> cave varied<br />
from -11 °C to 15°C, while <strong>the</strong> mean minimum<br />
internal temperature varied from<br />
3°C to 10°C. During <strong>the</strong> same period<br />
maximum temperatures varied from 10°C<br />
to 37°C outside and from 8°C to 11°C<br />
inside. The highest variation was<br />
observed for Te 2 (mean = 13.129; SD =<br />
7.467) followed by Te 1 (mean = 12.000;<br />
SD = 3.949), Ti 3 (mean = 9.574; SD =<br />
1.878), Ti 1 (mean = 8.973; SD = 1.489),<br />
and Ti 2 (mean = 8.914; SD = 1.269).<br />
Inside <strong>the</strong> cave <strong>the</strong> relative humidity varied<br />
from 80% to 100%, while outside it<br />
Figure 2. Light intensity variations recorded inside <strong>the</strong> cave (deep zone)<br />
VD r-- oo<br />
« •— — — — cs cs<br />
Distance <strong>of</strong> <strong>the</strong> sampling point from <strong>the</strong> entrance (m)<br />
• 12-Mar-OO: overcast sky -02-Jun-OO: clear sky
122 Biota 3/1-2,2002 PASTORELLI, LAGHI & SCARAVELLI<br />
fluctuated between 30 % and 100%. The<br />
highest variation was recorded for URe 2<br />
(mean = 68.714; SD = 21.504) followed<br />
by URe 1 (mean = 78.579; SD = 15.222),<br />
URi 3 (mean = 94.389; SD = 4.680), URi<br />
2 (mean = 94.762; SD = 4.230), and URi<br />
1 (mean = 95.143; SD = 3.525).<br />
Light intensity decreased to about 10 lux<br />
at 7 metres inside <strong>the</strong> cave, <strong>the</strong>n it gradually<br />
reached 0.0 lux at 21 m from <strong>the</strong><br />
entrance (Figure 2).<br />
Seasonal activity<br />
The total number <strong>of</strong> salamanders captured<br />
was 204 in <strong>the</strong> year 1999 and 181<br />
in <strong>the</strong> year 2000.<br />
The highest number <strong>of</strong> animals was captured<br />
in May (74 and 45 specimens in<br />
1999 and 2000 respectively) and<br />
September (45 and 36 specimens in 1999<br />
and 2000 respectively). The lowest num-<br />
ber <strong>of</strong> active salamanders was recorded in<br />
December 2000 and January 2001 (only<br />
3 active specimens in both cases) and no<br />
salamanders were found in January 2000.<br />
This activity pattern did not differ significantly<br />
between <strong>the</strong> two years (Mann-<br />
Whitney test T: 135.5 and P: 0.854).<br />
Activity <strong>of</strong> females, males, and both sexes<br />
pooled did not vary meaning<strong>full</strong>y<br />
between <strong>the</strong> two years (Mann-Whitney<br />
test respectively with P>0.5 in all cases).<br />
Salamanders activity was correlated with<br />
environmental parameters recorded during<br />
each sampling session. The number <strong>of</strong><br />
active juveniles, females, males, and<br />
adults was not significantly correlated to<br />
<strong>the</strong> relative humidity recorded inside and<br />
outside <strong>the</strong> cave (Spearman's correlation<br />
coefficient with P > 0.05 in all cases). The<br />
abundance <strong>of</strong> active salamanders was<br />
significantly correlated to internal tern-<br />
Figure 3. Regression between number <strong>of</strong> specimens captured and temperature measured<br />
7 meters far from <strong>the</strong> entrance.<br />
8 _ 9 . 1
PASTORELLI, LAGHI & SCARAVELLI Biota 3/1-2,2002 123<br />
peratures (Figure 3), except for males, for<br />
which abundance was significantly correlated<br />
with Ti 2 and Ti 3 only (P < 0.05 in<br />
all cases). A meaningful correlation was<br />
also found between <strong>the</strong> number <strong>of</strong> active<br />
salamanders and external temperatures<br />
(P< 0.05) except for males (P > 0.05);<br />
juvenile abundance was also not significantly<br />
correlated to Te2 (P > 0.05).<br />
Spatial distribution<br />
Taking into account <strong>the</strong> whole body <strong>of</strong><br />
captures, <strong>the</strong> animals were found at a<br />
mean distance <strong>of</strong> 6.5 m from <strong>the</strong><br />
entrance, and 51.3% <strong>of</strong> captures were<br />
made within <strong>the</strong> first 7 m <strong>of</strong> cave extension.<br />
Only 5 captures (1.4%) were made<br />
1 m outside <strong>the</strong> cave and only one<br />
(0.3%) more than 25 metres inside. This<br />
distribution pattern did not vary qualitatively<br />
between <strong>the</strong> two years.<br />
Juveniles were found at a mean distance<br />
<strong>of</strong> 3.9 m from <strong>the</strong> entrance, and 88.5%<br />
<strong>of</strong> observations were made within <strong>the</strong><br />
first 7 m. Only 2.3% <strong>of</strong> observations (4<br />
captures) were recorded 1 m outside <strong>the</strong><br />
cave.<br />
The mean distance from <strong>the</strong> entrance <strong>of</strong><br />
adults was 8.6 m; 66.7% <strong>of</strong> observations<br />
were recorded within 4 to 12 m from <strong>the</strong><br />
entrance and only 8.5% were found<br />
within <strong>the</strong> first 4 m from <strong>the</strong> entrance.<br />
Only one adult salamander (0.6%) was<br />
found more than 25 m from <strong>the</strong> entrance<br />
and only one male specimen (0.6%) 1 m<br />
outside <strong>the</strong> cave.<br />
The mean distance from <strong>the</strong> entrance<br />
recorded for females was 8.6 meters,<br />
while it was 8.8 meters for males.<br />
Significant variations were recorded<br />
between female and male distribution in<br />
<strong>the</strong> year 1999 (P < 0.05), but not in 2000<br />
(P > 0.05) or when years were pooled (P<br />
> 0.05). Superficial activity outside <strong>the</strong><br />
cave was observed only in December<br />
1999 and November 2000, close to <strong>the</strong><br />
entrance, when temperatures inside and<br />
outside <strong>the</strong> cave were very similar. There<br />
Figure 4. Seasonal variations <strong>of</strong> spatial distribution. Standard errors are reported.<br />
13,0<br />
12,0<br />
11,0<br />
10,0<br />
9,0<br />
8,0<br />
7,0<br />
6,0<br />
5,0<br />
4,0<br />
3,0<br />
2,0<br />
',0<br />
0,0<br />
Spring Summer FaH Winter<br />
D Juveniles d Females D Males
124 Biota 3/i-a, 2002 PASTORELLI, LAGHI & SCARAVELLI<br />
was no significant correlation between<br />
<strong>the</strong> spatial distribution <strong>of</strong> females and<br />
males and light intensity. However, <strong>the</strong><br />
spatial distribution <strong>of</strong> all specimens captured<br />
inside <strong>the</strong> cave was meaning<strong>full</strong>y<br />
correlated to light intensity (Spearman's<br />
correlation coefficient, P < 0.01 in all<br />
cases except for females: P < 0.05).<br />
The highest mean distance from <strong>the</strong><br />
entrance was recorded in summer (7.6<br />
m), followed by fall (6.6 m), spring (6.4<br />
m), and winter (3.9 m) respectively. This<br />
pattern did not show variations between<br />
females and juveniles, while males were<br />
found deeper in <strong>the</strong> cave in spring (Figure<br />
4).<br />
DISCUSSION<br />
Environmental parameters<br />
Inside <strong>the</strong> study cave, environmental variations<br />
were lower than those recorded<br />
outside; <strong>the</strong>rmal and hygrometric differences<br />
between <strong>the</strong> various sampling<br />
points inside <strong>the</strong> cave were on average<br />
low and generally due to different microhabitat<br />
conditions. During <strong>the</strong> study period,<br />
relative humidity inside <strong>the</strong> cave<br />
remained high throughout <strong>the</strong> year.<br />
Because <strong>of</strong> <strong>the</strong> direction <strong>of</strong> <strong>the</strong> cave's<br />
exposure, light penetrated only <strong>the</strong> first<br />
part <strong>of</strong> <strong>the</strong> cavity; <strong>the</strong>n its intensity gradually<br />
decreased, becoming 0.0 at 21<br />
meters inside.<br />
Seasonal activity<br />
Speleomantes italicus hypogeal observable<br />
activity showed great variations during<br />
<strong>the</strong> year, with <strong>the</strong> highest number <strong>of</strong><br />
salamanders captured in May and<br />
September and <strong>the</strong> lowest in January and<br />
December. These data are in contrast<br />
with those recorded for Speleomantes<br />
strinatii in an artificial tunnel. In fact, this<br />
latter population was more active in July<br />
and August (Salvidio et al. 1994).<br />
Salamander activity pattern was similar<br />
between years, but in 2000 a smaller<br />
number <strong>of</strong> salamanders was captured.<br />
This could be due to <strong>the</strong> variation in environmental<br />
factors between years, or perhaps<br />
to human disturbance; fur<strong>the</strong>r data<br />
are needed to clarify this observation.<br />
Our data show a direct influence <strong>of</strong> inside<br />
temperatures on salamander activity;<br />
again, <strong>the</strong>se results appear different from<br />
those obtained for S. strinatii, in which<br />
<strong>the</strong> influence <strong>of</strong> internal temperatures<br />
seemed to be indirect, as <strong>the</strong> highest correlation<br />
coefficients were obtained for<br />
temperatures recorded outside <strong>the</strong> tunnel<br />
or near its entrance (Salvidio et al. 1994).<br />
Spatial distribution<br />
Plethodontid salamanders are usually<br />
found in <strong>the</strong> twilight tract <strong>of</strong> a cave near<br />
its entrance (Cimmaruta et al. 1999,<br />
Lanza 1946, 1999, Pastorelli et al. 2001,<br />
Salvidio et al. 1994). As already observed<br />
for S. strinatii (Salvidio et al. 1994), in S.<br />
italicus juveniles concentrate near <strong>the</strong><br />
entrance <strong>of</strong> <strong>the</strong> cave and adult specimens<br />
in deeper zones.<br />
The hypo<strong>the</strong>sis <strong>of</strong> a spatial segregation<br />
between juveniles and adults due to<br />
behavioural interference is persuasive,<br />
and it has already been demonstrated in<br />
<strong>the</strong> American plethodontid genus<br />
Desmognathus (Colley et al. 1989) and<br />
verified by Salvidio & Pastorino (2002)<br />
even for Speleomantes strinatii.<br />
The overall spatial distribution <strong>of</strong> salamanders<br />
was positively correlated with<br />
light intensity. According to Roth (1976),<br />
Speleomantes are able to feed in complete<br />
darkness using only chemical cues.<br />
Notwithstanding this statement, is not<br />
unlikely that, when possible, cave salamanders<br />
prefer to forage in <strong>the</strong> twilight<br />
by means <strong>of</strong> visually guided prey catching<br />
behaviour.<br />
Similar to S. strinatii (Salvidio et al. 1994),<br />
even in a S. italicus population external<br />
activity was scarce and in proximity to <strong>the</strong><br />
cave entrance; <strong>the</strong> poor leaf soil produced<br />
by <strong>the</strong> coppiced wood near <strong>the</strong> cave<br />
probably cannot provide enough mois-
PASTORELLI, LAGHI & SCARAVELLI Biota 3/i-a, 2002 125<br />
ture to allow salamanders to move outside,<br />
except during very humid periods.<br />
Salamander distribution inside <strong>the</strong> cave<br />
varies seasonally, as already observed by<br />
Lanza (1946). In winter, when external<br />
conditions are similar to internal ones,<br />
salamanders are active at a short distance<br />
from <strong>the</strong> entrance, while in fall and spring<br />
<strong>the</strong> animals are found at a greater distance.<br />
In summer <strong>the</strong> animals are active<br />
deeper inside <strong>the</strong> cave, where environmental<br />
variations are lower.<br />
Acknowledgements<br />
The authors thank Benedetto Lanza and Sebastiano Salvidio for useful suggestions.<br />
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(Amphibia: Caudata: Plethodontidae). Atti del III Congresso Nazionale S.H.I.,<br />
Pianura 13: 83-84.<br />
MUTZ, T. 1998: Haltung und Zuchtdes Sardischen Hohlensalamanders Hydromantes imperialis<br />
(Stefani, 1969) und einige Beobachtungen zur Okologie der Europaischen<br />
Hohlensalamander. Salamandra 34: 167-180.<br />
NASCETTI, G., CIMMARUTA, R., LANZA, B. & BULLINI L. 1996: Molecular Taxonomy <strong>of</strong><br />
European Plethodontid Salamanders (Genus Hydromantes). Journal <strong>of</strong><br />
Herpetology 30: 161-183.<br />
PASTORELLI, C, LAGHI, P. & SCARAVELLI, D. 2001: Studi preliminari sull'ecologia di<br />
Speleomantes italicus (Dunn, 1923) nell'Appennino Tosco-Romagnolo. Atti del<br />
III Congresso Nazionale SHI (Pavia, 2000), Pianura 13: 347-351.<br />
ROTH G., 1976: Experimental analysis <strong>of</strong> <strong>the</strong> prey catching behaviour <strong>of</strong> Hydromantes italicus<br />
Dunn (Amphibia, Plethodontidae). J. Comp. Physiol. 109: 47-58<br />
SALVIDIO, S. 1990: Regime alimentaire d'une population epigee de Speleomantes ambrosii<br />
(Caudata, Plethodontidae) de la Ligurie Centrale (Italie septentrionale). Bulletin<br />
de la Societe Herpetologique de France 54: 69-72.<br />
SALVIDIO, S. 1991: Habitat ed attivita stagionale delle popolazioni interstiziali di<br />
Speleomantes ambrosii nell'alta Val Bisagno (Liguria Centrale). Rivista
126 Biota 3/1-2,2002 PASTORELLI, LAGHI & SCARAVELLI<br />
Piemontese di Storia Naturale 12: 69-74.<br />
SALVIDIO, S. 1992: Diet and food utilization in a rock-face population <strong>of</strong> Speleomantes<br />
ambrosii (Amphibia, Caudata, Plethodontidae). Vie Milieu 44: 35-39.<br />
SALVIDIO, S. 1993a: Life history <strong>of</strong> <strong>the</strong> European Plethodontid Salamander Speleomantes<br />
ambrosii (Amphibia, Caudata). Herpetological Journal 3: 55-59.<br />
SALVIDIO, S. 1993b: Struttura di popolazione del geotritone Speleomantes ambrosii. Suppl<br />
Ric. Biol. Selvaggina XXI: 517-520.<br />
SALVIDIO, S.r LATTES, A., TAVANO, M., MELODIA, F. & PASTORINO, M. V. 1994: Ecology<br />
<strong>of</strong> a Speleomantes ambrosii population inhabiting an artificial tunnel. Amphibia-<br />
Reptilia, 15:35-45.<br />
SALVIDIO, S. 1996: L'ecologia del Pletodontidi europei: stato delle ricerche sul geotritone<br />
Speleomantes ambrosii. Atti del 1 ° Convegno italiano di Erpetologia montana.<br />
Studi trentini di Scienze Naturali, Acta Biologica 71: 133-136.<br />
SALVIDIO, S. 1998: Estimating abundance and biomass <strong>of</strong> a Speleomantes strinatii<br />
(Caudata, Plethodontidae) population by temporary removal sampling.<br />
Amphibia-Reptilia 19: 113-124.<br />
SALVIDIO, S. & PASTORINO, M.V. 2002: Spatial segregation in <strong>the</strong> European plethodontid<br />
Speleomantes strinatii in relation to age and sex. Amphibia-Reptilia 23: 505-<br />
510.<br />
THORN, R. 1969: Les salamandres d'Europe d'Asie et d'Afrique du Nord. Paul Lechevalier,<br />
Paris.<br />
VOESENEK, L. A. C. J., ROOY (VAN), P. T. J. C. & STRIJBOSCH, H. 1987: Some autoecological<br />
data on <strong>the</strong> Urodeles <strong>of</strong> Sardinia. Amphibia-Reptilia 8: 307-314.
PASTORELLI, LAGHI & SCARAVELLI Biota 3/1-2.2002 127<br />
Speleomantes antipredator<br />
strategies: a review and new<br />
observations<br />
Christian PASTORELLI1, Paolo LAGHI2 & Dino<br />
SCARAVELLI3<br />
Y Cerchia di S. Egidio 2205, 47023 Cesena (FC), Italy<br />
E-mail: pastorellic@libero.it<br />
2v. Bruno C. Garibaldi 22, 47100 Forli, Italy<br />
E-mail: spelerpes@libero.it<br />
3Riserva Naturale Orientata e Museo di Onferno, p. Roma 1, 47855 Gemmano (RN),<br />
Italy<br />
E-mail: rnoonf@tin.it<br />
Abstract<br />
In <strong>the</strong> genus Speleomantes some antipredator adaptations are known, such as aposematic<br />
coloration, noxious skin secretions, body elevation and tail undulation, and immobility.<br />
During a long-term ecological study <strong>of</strong> Italian Cave Salamander in <strong>the</strong> cave<br />
"Grotta del Tritone" in <strong>the</strong> nor<strong>the</strong>rn Apennine mountains (Forll-Cesena province,<br />
Emilia-Romagna region, Italy), <strong>the</strong> authors observed some adult Speleomantes italicus<br />
displaying biting behaviour when taken by hand or forceps at <strong>the</strong> tail tip. Biting, as a<br />
defensive behaviour, is known for a number <strong>of</strong> plethodontid salamanders such as<br />
Aneides, Desmognathus, Gyrinophilus and Plethodon, but it has never before been<br />
observed and described in European plethodontid salamanders. Moreover, we observed<br />
<strong>the</strong> first case <strong>of</strong> damage to human skin by tail base gland secretions <strong>of</strong> Speleomantes<br />
supramontis, supporting <strong>the</strong> hypo<strong>the</strong>sis <strong>of</strong> a defensive role for <strong>the</strong> tail base glands in <strong>the</strong><br />
genus Speleomantes.<br />
Key words: Antipredator strategies, defensive biting behaviour, noxious skin secretions,<br />
Speleomantes, Plethodontidae.<br />
Received 3 August 2002; accepted 26 September 2002
128 Biota 3/1-a, -M PASTORELLI, LAGHI & SCARAVELLI<br />
INTRODUCTION<br />
Prey may respond evolutionary to predator<br />
pressure ei<strong>the</strong>r by removing <strong>the</strong>mselves<br />
from <strong>the</strong> foraging microhabitat <strong>of</strong><br />
<strong>the</strong> predators (predator avoidance mechanisms)<br />
or by reducing <strong>the</strong> probability <strong>of</strong><br />
successful predation when <strong>the</strong>y are within<br />
<strong>the</strong> perceptual field <strong>of</strong> <strong>the</strong> predators<br />
(antipredator mechanisms) (Brodie et al.<br />
1991). Antipredator mechanisms are<br />
widely found in plethodontid salamanders.<br />
Aposematic coloration, noxious skin<br />
secretions, body elevation, tail undulation,<br />
immobility, biting, rapid escape<br />
movements, tail autotomy, and o<strong>the</strong>r<br />
mechanisms have been extensively studied<br />
among <strong>the</strong> American members <strong>of</strong> this<br />
family (Brodie 1977, 1983, Brodie et al.<br />
1989, Garcia-Paris & Deban 1995,<br />
Labanick 1984). However, antipredator<br />
strategies <strong>of</strong> European plethodontid<br />
Speleomantes are still poorly known.<br />
MATERIALS AND METHODS<br />
A long-term ecological study <strong>of</strong> Italian<br />
Cave Salamander has been carried out<br />
since 1998 in <strong>the</strong> cave "Grotta del<br />
Tritone" [43°53'52" N - 0°29'02"W<br />
(Rome)], in <strong>the</strong> nor<strong>the</strong>rn Apennine<br />
mountains (Forli-Cesena province, Emilia-<br />
Romagna region, Italy). The aim <strong>of</strong> this<br />
research is to investigate severa aspects<br />
<strong>of</strong> <strong>the</strong> life history <strong>of</strong> Speleomantes italicus,<br />
such as seasonal activity, spatial distribution,<br />
individual displacements, population<br />
structure and size, diet, growth<br />
rates and reproductive behaviour<br />
(Pastorelli et al. 2001). During field work<br />
<strong>the</strong> authors observed some adult<br />
Speleomantes italicus displaying biting<br />
behaviour when taken by hand or forceps<br />
at <strong>the</strong> tail tip. This defensive behaviour<br />
Figure 1. The observed sequence <strong>of</strong> <strong>the</strong> defensive biting behaviour displayed by<br />
Speleomantes italicus (explanation in <strong>the</strong> text).
PASTORELLI, LAGHI & SCARAVELLI Biota 3/i-a, 2002 129<br />
was filmed with a digital video camera.<br />
During ano<strong>the</strong>r sampling session conducted<br />
in Sardinia in 1999, <strong>the</strong> authors<br />
observed <strong>the</strong> first case <strong>of</strong> damage to<br />
human skin by tail base gland secretions<br />
<strong>of</strong> Speleomantes supramontis.<br />
RESULTS AND DISCUSSION<br />
The biting behaviour was observed in<br />
four Speleomantes italicus adult females.<br />
(A) The animal, caught at <strong>the</strong> tail tip, was<br />
hanging from forceps or hands and rested<br />
immobile for a few seconds.<br />
(B) Then it tried to free itself with rapid<br />
coiling-uncoiling movements, sometimes<br />
while urinating or producing noxious skin<br />
secretions mainly from its tail base.<br />
(C: 1, 2, 3) The salamander <strong>of</strong>ten opened<br />
its mouth, and sometimes bit its own tail<br />
or <strong>the</strong> forceps.<br />
(D) After some seconds <strong>the</strong> stressed animal<br />
returned to its initial position and<br />
remained stationary or, more rarely,<br />
repeated <strong>the</strong> biting sequence after a<br />
while.<br />
Biting as defensive behaviour is known<br />
for a number <strong>of</strong> plethodontid salamanders,<br />
such as Aneides, Desmognathus,<br />
Gyrinophilus and Plethodon (Brodie et al.<br />
1989), but it has never before been<br />
described for European plethodontid<br />
salamanders. Desmognathus quadramaculatus<br />
use biting to repulse attacks <strong>of</strong><br />
snakes, such as Thamnophis sirtalis<br />
(Brodie etal. 1989). Speleomantes italicus<br />
may take some advantage by biting<br />
behaviour against Anguis fragilis and<br />
Natrix snakes (Lanza, 1999a). Also, <strong>the</strong><br />
adhesive nature <strong>of</strong> skin secretions <strong>of</strong> Cave<br />
Salamanders can be used against snakes,<br />
as quoted for Plethodon and Ensatina by<br />
Arnold (1982).<br />
Only adult females displayed <strong>the</strong> biting<br />
behaviour. Although <strong>the</strong>se are preliminary<br />
data, <strong>the</strong> hypo<strong>the</strong>sis <strong>of</strong> a correlation<br />
between biting and nest defence in<br />
Speleomantes should be taken into<br />
account and verified, as it has been<br />
experimentally demonstrated in some<br />
American plethodontids (Bachmann<br />
1984, Horn etal. 1990).<br />
Body coiling has been also observed in<br />
several plethodontids, such as Aneides,<br />
Batrachoseps, Desmognathus, Ensatina,<br />
Gyrinophilus, Hydromantes, Plethodon,<br />
Pseudotriton, and Bolitoglossa (Garcia-<br />
Paris & Deban 1995). In Hydromantes<br />
platycephalus, which is closely related to<br />
Speleomantes, this generalized escape<br />
strategy, associated with tucking limbs<br />
close to <strong>the</strong> body, results in a peculiar<br />
rolling antipredator escape behaviour,<br />
similar to that displayed by <strong>the</strong> anuran<br />
genus Oreophrynella; this may represent<br />
a convergence between two distantly<br />
related, taxa that both occur on rocky<br />
slopes (Garcia-Paris & Deban 1995). A<br />
similar antipredator behaviour was also<br />
observed in Speleomantes italicus.<br />
Indeed, this species displayed limb tucking<br />
when coiling as a consequence <strong>of</strong> disturbance,<br />
and sometimes dropped down<br />
from <strong>the</strong> cave walls to <strong>the</strong> floor; thus, <strong>the</strong><br />
occurrence <strong>of</strong> rolling escape or falling<br />
escape strategies in this genus should be<br />
verified. In S. italicus we also observed<br />
coiling, abundant skin secretions in <strong>the</strong><br />
tail base region, immobility (Lanza<br />
1999a), and escape by rapid snake-like or<br />
coiling-uncoiling movements.<br />
During sampling session, conducted by<br />
<strong>the</strong> authors in Sardinia in spring 1999,<br />
several specimens <strong>of</strong> all four Sardinian<br />
Speleomantes species were observed. In<br />
Speleomantes imperialis <strong>the</strong> authors<br />
observed abundant skin secretions at <strong>the</strong><br />
tail base region, body coiling, body elevation,<br />
and tail undulation, similar to <strong>the</strong><br />
antipredator strategies reported in Lanza<br />
(1999a).<br />
During <strong>the</strong> handling <strong>of</strong> a Speleomantes<br />
supramontis adult specimen that was<br />
taken by forceps at <strong>the</strong> trunk region, its<br />
tail base secretions mixed with urine were<br />
accidentally sprayed on <strong>the</strong> eyelid <strong>of</strong> one<br />
<strong>of</strong> <strong>the</strong> authors, causing irritation and
130 Biota PASTORELLI, LAGHI & SCARAVELLI<br />
scald-like dermatological symptoms<br />
(Figure 2). Human skin responses to salamander<br />
toxins began only few seconds<br />
after <strong>the</strong> contact and lasted about ten<br />
days. It is worth noting that absorption <strong>of</strong><br />
salamander venom was reduced by<br />
promptly washing <strong>the</strong> eyelid with fresh<br />
water.<br />
The skin secretions <strong>of</strong> plethodontid salamanders<br />
are known to be very irritating<br />
to human mucous membranes. Hansen<br />
(1990) reported a severe reaction, includ-<br />
ing temporary blindness, suffered by a<br />
human after handling an adult<br />
Hydromantes platycephalus. Toxicology<br />
<strong>of</strong> <strong>the</strong>se secretions has been studied only<br />
for S. italicus (Benedicenti & Polledro<br />
1899) and S. strinatii (Ph\sa\\x 1918). This<br />
is <strong>the</strong> first observed case <strong>of</strong> damage to<br />
human skin by tail base gland secretions<br />
(although mixed with urine) <strong>of</strong> S. supramontis,<br />
supporting <strong>the</strong> hypo<strong>the</strong>sis <strong>of</strong> its<br />
effective defensive function (Brizzi et al.<br />
1991).<br />
Figure 2. Human skin's reaction to Speleomantes supramontis tail base secretion include<br />
scald-like symptoms (in <strong>the</strong> rectangle).<br />
Acknowledgements<br />
We thank Pr<strong>of</strong>. Benedetto Lanza for useful suggestions and literature, Franca Monti and<br />
Luciano Cicognani who realized <strong>the</strong> digital film, and ST.E.R.N.A. company for technical<br />
support.<br />
REFERENCES<br />
ARNOLD, S. J. 1982: A quantitative approach to antipredator performance: Salamander<br />
defence against snake attack. Copeia 1982: 247-253.
PASTORELLI, LAGHI & SCARAVELLI Biota 3/1-2.2002 131<br />
BACHMANN, M. D. 1984: Defensive behaviour <strong>of</strong> brooding female red-backed salamanders<br />
(Plethodon cinereus). Herpetologica 40: 436-443.<br />
BENEDICENTI, A. & POLLEDRO ,O. 1899: Sulla natura e sulla azione fisiologica del veleno<br />
dello Spelerpes fuscus. Atti della regia Accademia dei Lincei, 296 Rendiconti 8<br />
(1st semester): 413-418.<br />
BRIZZI, R.r CALLONI, C. & DELFINO, G. 1991: Tail base glands in European plethodontid<br />
salamanders, with some comments on <strong>the</strong>ir biological and phylogenetic significance.<br />
Amphibia-Reptilia 12: 357-372.<br />
BRODIE, E. D. JR. 1977: Salamander antipredator postures. Copeia 1977: 523-535.<br />
BRODIE, E. D. JR 1983: Antipredator adaptations <strong>of</strong> salamanders: evolution and convergence<br />
among terrestrial species. In: Margaris N. S., Arianoutsou-Faraggitaki M. &<br />
Reiter R. J. (eds). Plant, animal, and microbial adaptations to terrestrial environment.<br />
Plenum Publishing Corporation; New York.<br />
BRODIE, E. D. JR., DOWDEY, T. G. & ANTHONY, C. 1989: Salamander antipredator strategies<br />
against snake attack: biting by Desmognathus. Herpetologica 45:167-171.<br />
BRODIE, E. D. JR., FORMANOWICZ, D. R. JR. & BRODIE III, E: D. 1991: Predator avoidance<br />
and antipredator mechanisms: distinct pathways to survival. Ethology<br />
Ecology & Evolution 3: 73-77.<br />
GARCIA-PARIS, M. & DEBAN, S. M. 1995: A novel antipredator mechanism in salamanders:<br />
rolling escape in Hydromantes platycephalus. Journal <strong>of</strong> Herpetology 29: 149-<br />
151.<br />
HANSEN, R. W. 1990: Hydromantes platycephalus (Mount Lyell Salamander). Toxicity.<br />
Herpetological Review 21: 91.<br />
HOM, C. L, WILLITS, N. H. & CLARK, C. W. 1990: Fitness consequences <strong>of</strong> nest defence in<br />
plethodontid salamanders: predictions <strong>of</strong> a dynamic optimisation model.<br />
Herpetologica 46: 304-319.<br />
LABANICK, G. M. 1984: Anti-predator effectiveness <strong>of</strong> autotomized tails <strong>of</strong> <strong>the</strong> salamander<br />
Desmognathus ochrophaeus. Herpetologica 40: 110-118.<br />
LANZA, B. 1999a: Speleomantes ambrosii (Lanza, 1955) - Ambrosis Holensalamander. In:<br />
Grossenbacher K. & Thiesmeier B. (eds). Handbuck der Reptilien und Amphibien<br />
Europas, Band 4/1, Schwanzlurche (Urodela) I (Hynobiidae, Proteidae,<br />
Plethodontidae, Salamandridae I: Pleurodeles, Salamandrina, Euproctus,<br />
Chioglossa, Mertensiella). AULA Verlag, Wiesbaden: 91-135.<br />
LANZA, B. 1999b: Speleomantes imperialis (Stefani, 1969) - Duftender Holensalamander.<br />
In: Grossenbacher K. & Thiesmeier B. (eds). Handbuck der Reptilien und<br />
Amphibien Europas, Band 4/1, Schwanzlurche (Urodela) I (Hynobiidae,<br />
Proteidae, Plethodontidae, Salamandridae I: Pleurodeles, Salamandrina,<br />
Euproctus, Chioglossa, Mertensiella). AULA Verlag, Wiesbaden: 155-163.<br />
PASTORELLI, C., LAGHI, P. & SCARAVELLI, D. 2001: Studi preliminari sull'ecologia di<br />
Speleomantes italicus (Dunn, 1923) nell'Appennino Tosco-Romagnolo. Atti del<br />
III Congresso Nazionale SHI (Pavia, 2000). Pianura 13: 347-351.<br />
PHISALIX, M. 1918: Les venins cutanes du Spelerpes fuscus Gray. Bulletin du Museum<br />
national d'Histoire naturelle 24: 92-96.<br />
STEFANI, R. 1969: La distribuzione geografica e I'evoluzione del geotritone sardo<br />
(Hydromantes genei Schleg.) e del geotritone continentale europeo<br />
(Hydromantes italicus Dunn). Archo zoologico italiano 53: 207-243.
POLYNOVA & POLYNOVA Biota 3/1-2.2002 133<br />
Tail autotomy as an index <strong>of</strong><br />
human influence on <strong>the</strong><br />
AlsophySax pipiens population in<br />
<strong>the</strong> Bogdino-Baskunchak state<br />
reserve<br />
Galina V. POLYNOVA1 & Olga E. POLYNOVA2<br />
'Russian Peoples' Friendship University, Department <strong>of</strong> Ecology, Pavlova 8/5, Moscow,<br />
113093, Russia<br />
2Moscow State University, Department <strong>of</strong> Geography, Vorobyovy Gory, MOSCOW,<br />
119899, Russia<br />
E-mail: olgapol@aport.ru<br />
Abstracts<br />
We started our investigations in <strong>the</strong> Alsophylax pipiens Pall, population in August 1998<br />
and we concluded it in August 2000. The first part <strong>of</strong> our research dealt with <strong>the</strong> undisturbed<br />
part <strong>of</strong> <strong>the</strong> population, and <strong>the</strong> second in a place where human influence is very<br />
high because <strong>of</strong> frequent excursions around <strong>the</strong> reserve territory. We collected <strong>the</strong> main<br />
population data in both territories and our comparison gave interesting results. We<br />
compared several sex-age groups in both parts <strong>of</strong> <strong>the</strong> population and counted <strong>the</strong> percentage<br />
<strong>of</strong> tail autotomy in each group. In almost all groups in <strong>the</strong> disturbed part <strong>of</strong> <strong>the</strong><br />
population this percentage is high. Therefore, tail autotomy can be an index <strong>of</strong> human<br />
pressure on ecosystems.<br />
Keywords: population, sex-age groups, tail autotomy<br />
Received 15 March 2002; accepted 3 August 2002
134 Biota 3/1-2,2002 POLYNOVA & POLYNOVA<br />
INTRODUCTION<br />
Alsophylax pipiens Pall, is <strong>the</strong> only species<br />
<strong>of</strong> <strong>the</strong> Gekkonidae family inhabiting<br />
Russia. The whole <strong>of</strong> <strong>the</strong> species' area in<br />
Russia is represented by <strong>the</strong> territory <strong>of</strong><br />
<strong>the</strong> isolated population inhabiting <strong>the</strong><br />
Bolshoe Bogdo mountain in <strong>the</strong> nor<strong>the</strong>astern<br />
part <strong>of</strong> <strong>the</strong> Astrakhan region,<br />
which is situated beside a beach with <strong>the</strong><br />
largest salt deposits in <strong>the</strong> world, on <strong>the</strong><br />
shore <strong>of</strong> Lake Baskunchak. This is where<br />
Alsophylax pipiens was described for <strong>the</strong><br />
first time in 1813 by <strong>the</strong> great Russian scientist<br />
and traveller, Peter Simeon Pallas.<br />
In 1997 <strong>the</strong> environs <strong>of</strong> Lake Buskunchak<br />
and <strong>the</strong> mountain Bolshoe Bogdo<br />
acquired <strong>the</strong> status <strong>of</strong> <strong>the</strong> Bogdino-<br />
Baskunchak state reserve.<br />
Our investigations <strong>of</strong> <strong>the</strong> Alsophylax pipiens<br />
population started in 1995 as <strong>the</strong><br />
part <strong>of</strong> <strong>the</strong> Astrakhan Herpetological<br />
expedition, financed by <strong>the</strong> MacArthur<br />
and ISAR Foundations (Polynova &<br />
Bozshansky 1995, 1998a, 1998b,<br />
Bozshansky & Polynova 1997,<br />
Bozshansky & Polynova 1998, Polynova<br />
& Polynova 2000). We continued our<br />
work in August 1998 and in August 2000<br />
with <strong>the</strong> Student Ecological Expedition <strong>of</strong><br />
Figure 1. Size by groups (1-5) <strong>of</strong> males and subadults (1998).<br />
«<br />
6-,<br />
5 -<br />
I ^<br />
B 3<br />
d o<br />
Z z "<br />
1 -<br />
Q<br />
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•<br />
.<br />
• - -<br />
' • :<br />
5<br />
*,<br />
i<br />
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11<br />
j<br />
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3<br />
<strong>the</strong> Russian Peoples' Friendship<br />
University, Ecological Department. The<br />
main purpose <strong>of</strong> <strong>the</strong> expedition was to<br />
investigate human influence on <strong>the</strong><br />
reserve's ecosystems. The mountain<br />
Bolshoe Bogdo is <strong>the</strong> centre <strong>of</strong> <strong>the</strong> recreational<br />
burden <strong>of</strong> <strong>the</strong> reserve's territory.<br />
This article deals with <strong>the</strong> data ga<strong>the</strong>red<br />
during <strong>the</strong> 1998 and 2000 field seasons<br />
and presents some interesting results.<br />
MATERIAL AND METHODS<br />
We used <strong>the</strong> following methods: catching,<br />
measuring, sex determining, marking<br />
with colour and finger cutting <strong>of</strong>f; mapping<br />
<strong>of</strong> lizards' meetings and movements;<br />
biotopes description and daily activity<br />
registration. The last two data will form<br />
<strong>the</strong> basis <strong>of</strong> fur<strong>the</strong>r publications.<br />
The young lizards that we couldn't distinguish<br />
by sex rate were put into <strong>the</strong><br />
subadult group (Figure 1, group no. 1).<br />
All <strong>of</strong> <strong>the</strong>m spent one winter. We based<br />
our division <strong>of</strong> adult males and females<br />
into size-age groups on <strong>the</strong>ir body length.<br />
It is well known that adult lizards grow<br />
very slowly. That is why <strong>the</strong> variation line<br />
<strong>of</strong> <strong>the</strong>ir body length is continuous.<br />
Never<strong>the</strong>less, <strong>the</strong> male data from 1998<br />
?<br />
\<br />
s<br />
><br />
J<br />
"<br />
4<br />
,<br />
"<br />
-••H<br />
|" ' - -<br />
-<br />
. .<br />
t<br />
i K<br />
t °<br />
.< ^^<br />
21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41<br />
Body size (mm)
POLYNOVA & POLYNOVA Biota 3/1-2,2002 135<br />
Figure 2. Size groups <strong>of</strong> females (1998)<br />
34 -^' 35 -" 36 -: !-37, •*•*' 38 39<br />
*«,, ";v Body size (mm)<br />
show markedly distinguished groups<br />
(Figure 1, no. 2-5). The division <strong>of</strong> female<br />
groups is based on <strong>the</strong>ir variation line<br />
(Figure 2) and by analogy with <strong>the</strong> distinguished<br />
size groups <strong>of</strong> males. This<br />
method is imperfect to some extent, but<br />
Table 1. Body length <strong>of</strong> sex and age groups (public data)<br />
Sex and age groups<br />
Juveniles<br />
Juveniles<br />
Juveniles<br />
Subadults<br />
Subadults<br />
Just adults<br />
Just adults<br />
Just adults<br />
Adult males<br />
Adult males<br />
Adult females<br />
Adult females<br />
Body length (mm)<br />
16-18<br />
15-17<br />
18-20<br />
20-26.6 (22.41±0.60)<br />
19-24 (21. 5±0.2)<br />
25-28<br />
31-32<br />
34<br />
33-36 (35.0±0.3)<br />
22.5-33-0(31.310.5)<br />
31-33 (33.110.5)<br />
30-40(36.110.5)<br />
as <strong>the</strong> species is so small and rare, we<br />
considered it to be impossible to investigate<br />
lizards' age by <strong>the</strong> classic methods <strong>of</strong><br />
comparing annual bone rings. Table 1<br />
shows public data on this species' body<br />
length. The differences between subadult<br />
Authors<br />
Scherbak & Golubev, 1986<br />
Smirnovetal, 1985<br />
Chernov 1947<br />
Brushko 1995<br />
Shammakov 1981<br />
Andrushkol9555 Scherbak<br />
& Golubev 1986<br />
Brushko 1995<br />
Smirnovetal. 1985<br />
Kubykin 1975<br />
Brushko 1995<br />
Brushko 1995<br />
Kubykin 1975
136 Biota 3/i-a, 2002 POLYNOVA & POLYNOVA<br />
Table 2. Sex-age groups and tail autotomy percentage (August 1998).<br />
No. <strong>of</strong><br />
group<br />
1<br />
2<br />
3<br />
4<br />
5<br />
6<br />
7<br />
8<br />
Sex and age<br />
Subadults<br />
S3<br />
33<br />
•/ *//v5
POLYNOVA & POLYNOVA Biota 3/1-2,2002 137<br />
Figure 4. Size by groups (4-7) <strong>of</strong> females (2000).<br />
females (Figure 1,2). The average sizes <strong>of</strong><br />
groups are presented in Table 2. O<strong>the</strong>r<br />
authors (Scherbak & Golubev 1986) also<br />
show <strong>the</strong> presence <strong>of</strong> several sex-age<br />
groups in this species.<br />
The average male/female sex-ratio is<br />
1.4:1. The same-size male/female sexratio<br />
(groups no. 4 and no. 6) is 2:1. The<br />
older sex groups do not coincide, but on<br />
<strong>the</strong> whole we see <strong>the</strong> prevalence <strong>of</strong><br />
females over males. The prevalence <strong>of</strong><br />
males in <strong>the</strong> whole population is due to<br />
young adult males (groups no. 2 and no.<br />
3). These males are probably from two<br />
clutches <strong>of</strong> <strong>the</strong> same year. We base this<br />
probability on <strong>the</strong> information about two<br />
clutches known in this species (Scherbak<br />
& Golubev 1986).<br />
We see that in <strong>the</strong> youngest part <strong>of</strong> <strong>the</strong><br />
population <strong>the</strong>re is a prevalence <strong>of</strong> males,<br />
<strong>the</strong>n <strong>the</strong> sex-ratio changes to 1:1. As time<br />
goes by, <strong>the</strong> number <strong>of</strong> <strong>the</strong> oldest females<br />
prevails over <strong>the</strong> number <strong>of</strong> <strong>the</strong> oldest<br />
males. Our data are quite representative,<br />
because we took <strong>the</strong>m from <strong>the</strong> wide territory<br />
<strong>of</strong> <strong>the</strong> population.<br />
The next body data - <strong>the</strong> length <strong>of</strong> <strong>the</strong> tail<br />
- needs special attention. It includes little<br />
information about <strong>the</strong> lizard's age<br />
because <strong>of</strong> <strong>the</strong> high tail autotomy percent,<br />
but it has ano<strong>the</strong>r value. Tail autotomy<br />
percentage increases from subadult<br />
animals (14%) to mature females (100%)<br />
(Table 2). This increase is quite natural:<br />
those lizards who enjoy a longer life have<br />
more accidents. There are some public<br />
data about tail autotomy levels in <strong>the</strong><br />
Alsophylax pipiens populations (16.7% -<br />
Shammakov 1981, 30% - Anan'eva &<br />
Muhtabayar 1997).<br />
The next stage <strong>of</strong> our investigations in<br />
August 2000 was devoted to <strong>the</strong> most<br />
disturbed part <strong>of</strong> <strong>the</strong> species' population.<br />
It lies in <strong>the</strong> nor<strong>the</strong>astern territory, at <strong>the</strong><br />
top <strong>of</strong> <strong>the</strong> mountain. There we caught,<br />
measured, and marked 70 lizards (Table<br />
3, Figure 3, 4).<br />
As we can see, <strong>the</strong> nor<strong>the</strong>astern part <strong>of</strong><br />
<strong>the</strong> population also has 7 sex-age groups:<br />
1 subadult group, 2 groups <strong>of</strong> adult males<br />
and 4 groups <strong>of</strong> adult females. Subadults<br />
were longer than in 1998, and adult<br />
males formed only two distinguished<br />
groups: <strong>the</strong> younger male group No. 2<br />
unites two groups from 1998 (no. 2, no.<br />
3, Table 2). The next size-age male group<br />
No. 3 was <strong>the</strong> same as in 1998 (No.4,<br />
Table 2). Finally, we couldn't find <strong>the</strong> oldest<br />
male group in 2000. At <strong>the</strong> same<br />
time, females also formed a continuous<br />
variation line, which we divided approximately<br />
into 4 groups (by analogy with<br />
previous data and due to several maximums).<br />
It was more than in 1998. We
138 Biota 3/i-a, 2002 POLYNOVA & POLYNOVA<br />
found younger females No.4, and o<strong>the</strong>r<br />
female groups were approximately <strong>the</strong><br />
same in both seasons.<br />
It is obvious that <strong>the</strong>re were some<br />
changes in <strong>the</strong> male/female sex-ratio.<br />
The average sex -ratio was 1.6:1 (males :<br />
females), <strong>the</strong> same-age youngest<br />
male/female sex - ratio was 4.3:1 (males<br />
no. 2 and females no. 4, Table 3), <strong>the</strong>n it<br />
changed to 1:1 (males no. 3 and females<br />
no. 5 and no. 6) and, finally, <strong>the</strong>re were<br />
no oldest males corresponding to <strong>the</strong> oldest<br />
females.<br />
We can now see that <strong>the</strong> data <strong>of</strong> 1998<br />
and 2000 demonstrate <strong>the</strong> prevalence <strong>of</strong><br />
males in <strong>the</strong> whole population. This is<br />
because <strong>of</strong> <strong>the</strong> prevalence <strong>of</strong> males in <strong>the</strong><br />
youngest group. Then we see <strong>the</strong> increasing<br />
proportion <strong>of</strong> females in older groups<br />
and <strong>the</strong> prevalence <strong>of</strong> females over males<br />
in <strong>the</strong> oldest part. We couldn't find such<br />
detailed information in public data. Some<br />
authors (Andrushko 1955, Scherbak &<br />
Golubev 1986) point out 1:1 sex -ratio in<br />
this species.<br />
Sex-age groups and sex-ratio data from<br />
<strong>the</strong> 1998 and 2000 field seasons have<br />
much in common, but tail autotomy<br />
materials differ. As we have shown, <strong>the</strong><br />
tail autotomy percentage <strong>of</strong> <strong>the</strong> undisturbed<br />
sou<strong>the</strong>astern population group<br />
naturally increases from <strong>the</strong> youngest to<br />
<strong>the</strong> oldest animals (Table 2). At <strong>the</strong> same<br />
time, <strong>the</strong> tail autotomy percentage <strong>of</strong> <strong>the</strong><br />
northwestern part is very high in all sexage<br />
rate groups, with exception <strong>of</strong> <strong>the</strong><br />
youngest females: no. 4 (Table 3). It is<br />
well known that tail autotomy in natural<br />
lizard populations can indicate natural<br />
predation pressure (Pianka 1970, Medel<br />
et. al., 1988 and o<strong>the</strong>rs), lizard density,<br />
aggressive interactions <strong>of</strong> competitive<br />
individuals, and o<strong>the</strong>r factors.<br />
The comparison <strong>of</strong> <strong>the</strong>se two population<br />
groups shows that: 1) <strong>the</strong> density <strong>of</strong><br />
lizards in <strong>the</strong> northwestern disturbed area<br />
is lower than in <strong>the</strong> sou<strong>the</strong>astern undisturbed<br />
area, so aggressive interactions<br />
between competitive individuals must be<br />
lower too; 2) natural predation (our<br />
observations) is <strong>the</strong> same; 3) <strong>the</strong> main<br />
difference between <strong>the</strong>se areas that can<br />
influence tail autotomy is <strong>the</strong> recreational<br />
burden. The high recreational burden <strong>of</strong><br />
this region consists <strong>of</strong> dozens <strong>of</strong> tourists<br />
and <strong>the</strong>ir cars. On weekends <strong>the</strong> number<br />
<strong>of</strong> visitors increases by three or four<br />
times. Lizards practically live under<br />
tourists' feet and our experience shows<br />
<strong>the</strong> ease <strong>of</strong> lizards' tail loss.<br />
The man-made pressure level can be<br />
undoubtedly measured for almost every<br />
animal and plant species that suffers it.<br />
But we know some indicator species that<br />
are most suitable for this aim. To our<br />
minds, <strong>the</strong> tail autotomy level in lizard<br />
populations may be one <strong>of</strong> <strong>the</strong>m. This<br />
index can be <strong>of</strong> value in factoring <strong>the</strong> disturbance<br />
caused by <strong>the</strong> recreational burden.<br />
We think that such an index is more<br />
important, especially in reserves, than<br />
investigations in already disturbed<br />
ecosystems, but it needs fur<strong>the</strong>r research.
POLYNOVA & POLYNOVA Biota 3/1-2,2002 139<br />
REFERENCES<br />
ANAN'EVA, N.B. & MUNHBAYAR, K. 1997: Pisklivyy gekkonchik. In: Zemnovodnye i presmykauschiesya<br />
Mongolii. Presmykauschiesya. Kollektivnaya monografiya.<br />
Sovmestnaya rossiysko-mongorskaya ekspediciya. RAN, AN Mongolii. KMK Ltd.<br />
M.: 14-27.<br />
ANDRUSHKO, A.M. 1955: Presmykauschiesya Kazahskogo nagor'ya i ih hozyaystvennoe<br />
znachenie. Uchen. zap. Leningr. un-ta. Ser. biol. nauk 181: 19-43.<br />
BOZHANSKIY, AT. & POLYNOVA, G.V. 1998: Proekt regional'nogo spiska reptiliy Krasnoy<br />
knigi Astrahanskoy oblasti. In: Problemy sohraneniya bioraznoobraziya aridnyh<br />
regionov Rossii. Materialy mezhdunarodnoy nauchno-prakticheskoy konferencii.<br />
Volgograd, Rossiya, 11-17 sentyabrya 1998: 57-59.<br />
BRUSHKO, Z.K. 1995: Yaschericy pustyn' Kazahstana. Izd-vo Konzhyk, Almaty: 231 p.<br />
KUBYKIN, R.A. 1975: Ekologo-faunisticheskiy obzor reptiliy ostrovov oz. Alakol' (Vost.<br />
Kazahstan). Izv. AN KazSSR. Ser. biol. 3: 10-16.<br />
POLYNOVA, G.V. & POLYNOVA, O.E. 2000: Problemy sohraneniya gerpet<strong>of</strong>auny<br />
Astrahanskoy oblasti. In: Aktual'nye problemy ekologii i prirodopoPzovaniya. Izdvo<br />
RUDN, M.: 65-70.<br />
SMIRNOV, S.I., SHKUNOV, V.F., KUDAKINA, E.I. 1985: Gekkony Severnogo Prikaspiya. In:<br />
Voprosy gerpetologii. Nauka, L.: 195-196.<br />
CHERNOV, S.A. 1947: Materialy k gerpet<strong>of</strong>aune Kazahskogo nagor'ya, severnogo<br />
poberezh'ya Balhasha i gor. Kin-Tau. Izv. AN KazSSR. Ser. zool., vyp. 6.: 120-124.<br />
SHAMMAKOV, S. 1981: Presmykauschiesya ravninnogo Turkmenistana. Ylym, Ashhabad:<br />
311 p.
RACCA Biota 3/1-2.2002 141<br />
The conservation <strong>of</strong> <strong>the</strong> Agile Frog<br />
Rana dalmatina in Jersey<br />
(Channel Islands)<br />
Laura RACCA<br />
The Durrell Institute <strong>of</strong> Conservation and Ecology, University <strong>of</strong> Kent, Canterbury, Kent,<br />
CT2 7NS, UK<br />
E-mail: Iauracca1@yahoo.it<br />
Abstract<br />
The Jersey population <strong>of</strong> <strong>the</strong> Agile Frog has been declining in both range and number<br />
since <strong>the</strong> early 1900s. At <strong>the</strong> present time, <strong>the</strong>re is only one site on <strong>the</strong> island that supports<br />
a natural agile frog population. Since <strong>the</strong> 1980s, a captive breeding, reintroduction<br />
and habitat management programme involving various organisations has been trying<br />
to arrest this dramatic decline in agile frog numbers in <strong>the</strong> wild. All parties involved<br />
agreed that without a clearer understanding <strong>of</strong> <strong>the</strong> ecology and habitat requirements <strong>of</strong><br />
<strong>the</strong> agile frog in Jersey, it was difficult to develop management techniques for <strong>the</strong> protection<br />
and improvement <strong>of</strong> key habitat areas. The objectives <strong>of</strong> <strong>the</strong> study are to (1)<br />
estimate <strong>the</strong> number <strong>of</strong> adult frogs through a mark - recapture programme; (2) monitor<br />
hatching success and recruitment; (3) study <strong>the</strong> interactions with o<strong>the</strong>r species present<br />
in <strong>the</strong> pond (toads, newts, invertebrates and grass snakes) and in <strong>the</strong> surrounding<br />
area (grass snakes, small mammals and birds); (4) determine <strong>the</strong> frog's overwintering<br />
preferences; (5) monitor <strong>the</strong> captive breeding programme and carry out an investigation<br />
<strong>of</strong> potential reintroduction sites; (6) develop optimal conditions for captive rearing<br />
<strong>of</strong> tadpoles and consider a health monitoring protocol prior to release <strong>of</strong> captive raised<br />
frogs; (7) compare <strong>the</strong> habitat requirements <strong>of</strong> <strong>the</strong> Agile Frog in Jersey and in nor<strong>the</strong>rn<br />
France. Preliminary results <strong>of</strong> <strong>the</strong> study obtained from <strong>the</strong> observations collected during<br />
<strong>the</strong> first fieldwork season are presented here.<br />
Key words: conservation, decline, ecology, Jersey, Rana dalmatina, Species Action Plan.<br />
Received 6 September 2001; accepted 24 October 2001
142 Biota 3 i-->, RACCA<br />
INTRODUCTION<br />
The island <strong>of</strong> Jersey is situated 21 kilometres<br />
<strong>of</strong>f <strong>the</strong> north-west coast <strong>of</strong> France<br />
and 128 kilometres from <strong>the</strong> English<br />
coast. Jersey is <strong>the</strong> largest <strong>of</strong> <strong>the</strong> Channel<br />
Islands with a surface area <strong>of</strong> 117 square<br />
kilometres and an <strong>of</strong>ficial population <strong>of</strong><br />
approximately 85,000. The island has its<br />
own government, called <strong>the</strong> States <strong>of</strong><br />
Jersey, which comprises 53 elected members.<br />
Jersey also has its own system <strong>of</strong><br />
local administration, fiscal and legal systems,<br />
and courts <strong>of</strong> law. Jersey is nei<strong>the</strong>r<br />
part <strong>of</strong> <strong>the</strong> United Kingdom nor a colony.<br />
It is not represented in <strong>the</strong> United<br />
Kingdom Parliament, whose Acts extend<br />
to Jersey only if <strong>the</strong> island expressly<br />
agrees that <strong>the</strong>y should do so. The island<br />
has allegiance to <strong>the</strong> British Crown. Jersey<br />
is not part <strong>of</strong> <strong>the</strong> European Union (States<br />
<strong>of</strong> Jersey 2001).<br />
The Environmental Services Unit (ESU) is<br />
<strong>the</strong> department within <strong>the</strong> States <strong>of</strong><br />
Jersey Planning and Environment<br />
Committee involved in almost every<br />
aspect <strong>of</strong> environmental management in<br />
Jersey. On <strong>the</strong> island, certain ecosystems<br />
are protected by a designation under <strong>the</strong><br />
planning law. They are called Sites <strong>of</strong><br />
Special Interest (SSI) and are monitored<br />
by <strong>the</strong> ESU to ensure that <strong>the</strong> conditions<br />
that make <strong>the</strong>m »special« are not lost<br />
(States <strong>of</strong> Jersey 2001).<br />
The following amphibians and reptiles are<br />
found in Jersey (Tonge 1986): Agile Frog<br />
Rana dalmatina Common Toad Bufo<br />
bufo, Palmate Newt Triturus helveticus,<br />
Grass Snake Matrix natrix, Slow Worm<br />
Anguis fragilis, Green Lizard Lacerta bilineata<br />
and Wall Lizard Podarcis muralis.<br />
They are all protected under schedule 1<br />
<strong>of</strong> <strong>the</strong> Conservation <strong>of</strong> Wildlife (Jersey)<br />
Law 2000 (Jersey Legal Information<br />
Board 2001).<br />
Figure 1. North Slack, Ouaisne' Common. The only site in <strong>the</strong> island that supports a<br />
wild population <strong>of</strong> Rana dalmatina. The terrestrial habitat around <strong>the</strong> slack is a developing<br />
heathland.
RACCA Biota 3/i-a, 2002 143<br />
DISTRIBUTION AND DECLINE OF THE<br />
AGILE FROG<br />
Apart from Jersey, <strong>the</strong> agile frog is not<br />
found anywhere else in <strong>the</strong> British Isles<br />
(Grossenbacher 1997). The Jersey population<br />
<strong>of</strong> <strong>the</strong> agile frog has been declining<br />
in both range and numbers since <strong>the</strong><br />
early 1900s (Gibson & Freeman 1997). In<br />
<strong>the</strong> 1970s, <strong>the</strong> frog could be found in<br />
only seven localities, and by <strong>the</strong> 1980s<br />
this had dropped to only two sites (Tonge<br />
1986). In 1987, a spill <strong>of</strong> aldicarb (an<br />
agricultural pesticide) in one <strong>of</strong> <strong>the</strong> sites<br />
caused <strong>the</strong> loss <strong>of</strong> one <strong>of</strong> <strong>the</strong> two remaining<br />
populations <strong>of</strong> frogs (Gibson &<br />
Freeman 1997). Therefore, at <strong>the</strong> present,<br />
<strong>the</strong>re is only one site in <strong>the</strong> island, a<br />
coastal heathland, which supports a population<br />
<strong>of</strong> Rana dalmatina in <strong>the</strong> wild<br />
(Figure 1).<br />
Possible causes <strong>of</strong> agile frog decline in<br />
Jersey are <strong>the</strong> following (Agile Frog<br />
Group 2001):<br />
1) Habitat loss / fragmentation. Jersey is<br />
a densely populated island and habitat<br />
fragmentation continues through development<br />
for housing and <strong>the</strong> associated<br />
services required. In addition, small-scale<br />
turnover <strong>of</strong> semi-natural habitats, including<br />
conversion <strong>of</strong> heathland and marginal<br />
farmland to agricultural land, mean<br />
that important habitat areas are continually<br />
being lost.<br />
2) Water quality. Jersey, not being part <strong>of</strong><br />
<strong>the</strong> European Union, does not have to<br />
follow its guidelines for <strong>the</strong> maintenance<br />
<strong>of</strong> water quality. In Jersey, pollution <strong>of</strong><br />
groundwater is caused by two main<br />
sources, agriculture and domestic wastes.<br />
Agricultural nutrients and sewage from<br />
defective or poorly located soakaways are<br />
implicated in <strong>the</strong> pollution <strong>of</strong> <strong>the</strong> groundwater<br />
by <strong>the</strong> abnormally high nitrate concentrations<br />
observed. In addition, pesticide<br />
residue has <strong>of</strong>ten been detected in<br />
run-<strong>of</strong>f from arable farmland.<br />
3) Pollution events. As previously mentioned,<br />
a spillage <strong>of</strong> aldicarb in Noirmont<br />
pond in 1987 had a lethal effect on many<br />
frog and toad eggs and adults, and basically<br />
halved <strong>the</strong> agile frog's range<br />
overnight. Ano<strong>the</strong>r pollution event was a<br />
recent contamination <strong>of</strong> an area close to<br />
<strong>the</strong> airport by run-<strong>of</strong>f from airport firefighting<br />
practice. In addition, <strong>the</strong>re are<br />
countless small scale pollution events<br />
occurring on a frequent basis throughout<br />
Jersey which threaten both aquatic and<br />
terrestrial habitats.<br />
4) Water shortage. Agriculture is <strong>the</strong> principal<br />
land use in Jersey and covers 58%<br />
<strong>of</strong> <strong>the</strong> total land area <strong>of</strong> <strong>the</strong> island. Land<br />
is intensively farmed due to <strong>the</strong> excellent<br />
soil quality, and <strong>the</strong> scarcity <strong>of</strong> land frequently<br />
leads to double cropping. The<br />
industry exacts a high demand from <strong>the</strong><br />
island's water supply and <strong>of</strong>ten improves<br />
land drainage to ensure rapid run-<strong>of</strong>f.<br />
Additionally, many houses are still not<br />
connected to a main water supply, and<br />
are instead reliant on <strong>the</strong> ground water.<br />
These agricultural and domestic factors<br />
may have caused <strong>the</strong> lowering <strong>of</strong> <strong>the</strong><br />
water table that is now being experienced<br />
in Jersey, which has resulted in <strong>the</strong> loss <strong>of</strong><br />
many ephemeral ponds.<br />
5) Predation pressures. Apart from <strong>the</strong>ir<br />
natural predators (palmate newts, some<br />
aquatic macro-invertebrates, small mammals,<br />
grass snakes, etc), frogs and toads<br />
are now also being predated by pets,<br />
which are increasing in numbers with <strong>the</strong><br />
growth in human population. Domestic<br />
cats and polecat-ferrets are both plentiful<br />
as feral pests. More recently, large numbers<br />
<strong>of</strong> feral ducks have invaded many<br />
available water bodies, including <strong>the</strong> agile<br />
frog's only natural breeding site.<br />
6) Small population effects. The Jersey<br />
population <strong>of</strong> <strong>the</strong> agile frog has, and still<br />
is, going through a severe Cbottleneckx<br />
effect. At <strong>the</strong> present time, <strong>the</strong> factors<br />
listed above are likely to be <strong>the</strong> most<br />
influential in determining <strong>the</strong> immediate<br />
survival <strong>of</strong> <strong>the</strong> agile frog. In <strong>the</strong> longer<br />
term, however, any surviving population
144 Biota 3/1-2,2002 RACCA<br />
could become exposed to genetic risk<br />
factors, such as inbreeding depression<br />
(Caughley 1994).<br />
CAPTIVE BREEDING PROGRAMME OF<br />
THE AGILE FROG IN JERSEY<br />
A collaborative programme incorporating<br />
captive-breeding, reintroduction and<br />
habitat management started in <strong>the</strong> late<br />
1980s in order to try to arrest <strong>the</strong> potentially<br />
terminal decline in agile frog numbers<br />
in Jersey. The organisations involved<br />
are <strong>the</strong> Environmental Services Unit<br />
(ESU), <strong>the</strong> Herpetology Department <strong>of</strong><br />
<strong>the</strong> Durrell Wildlife Conservation Trust,<br />
and <strong>the</strong> Zoology Section <strong>of</strong> <strong>the</strong> Societe<br />
Jersiaise. The main objectives <strong>of</strong> <strong>the</strong> programme<br />
are (Agile Frog Group 2001) (1)<br />
to maintain self-sustaining captive safety<br />
net populations in <strong>the</strong> event <strong>of</strong> <strong>the</strong> extirpation<br />
<strong>of</strong> <strong>the</strong> species from <strong>the</strong> remaining<br />
natural breeding site and (2) to generate<br />
surplus tadpoles and froglets for reintroduction<br />
into existing and potential new<br />
sites.<br />
Figure 2 shows <strong>the</strong> total number <strong>of</strong> egg<br />
clumps found in <strong>the</strong> wild and number <strong>of</strong><br />
clumps taken for <strong>the</strong> captive breeding<br />
programme from 1990 to 2001. It is not<br />
clear if <strong>the</strong> lack <strong>of</strong> clumps in <strong>the</strong> wild dur-<br />
ing 1994, 1995, and 1996 was real or if<br />
spawn were missed during <strong>the</strong> surveys<br />
because <strong>of</strong> <strong>the</strong> unavailability <strong>of</strong> surveyors.<br />
REINTRODUCTION AND INTRODUC-<br />
TION SITES<br />
Grosnez pond<br />
This site (Figure 3) has never supported<br />
populations <strong>of</strong> agile frogs. It was chosen<br />
as an introduction site because it had<br />
good quality water and was a breeding<br />
site for toads and newts (Agile Frog<br />
Group 2001). The introduction started in<br />
1994 and <strong>the</strong> first clump <strong>of</strong> frog eggs was<br />
found two years later. Breeding has reoccurred<br />
in 1998, 1999, and 2000.<br />
Tadpoles were released in <strong>the</strong> pond for<br />
<strong>the</strong> last time in 2000 (Racca 2000).<br />
Noirmont pond<br />
This site (Figure 3) was an historic breeding<br />
pond for <strong>the</strong> agile frog until 1987,<br />
when a spill <strong>of</strong> an agricultural pesticide<br />
polluted its water. It was chosen as a reintroduction<br />
site after <strong>the</strong> results <strong>of</strong> water<br />
analyses indicated that <strong>the</strong> pollution<br />
problem had been solved and after<br />
observing that introduced toad tadpoles<br />
developed success<strong>full</strong>y to metamorpho-<br />
Figure 2. Total number <strong>of</strong> egg clumps found in <strong>the</strong> wild and numer <strong>of</strong> clumps taken for<br />
<strong>the</strong> captive rearing programme, from 1990 to 2001.<br />
a, 12-]<br />
1 10-<br />
1 8<br />
? 4-<br />
-i 2 -<br />
i 0<br />
0 No <strong>of</strong> chimps found at Ouaisne'<br />
D No <strong>of</strong> clumps taken for captive rearing programme
RACCA Biota 3/1-2, 2002 145<br />
Figure 3. Grosnez pond (A) and enclosure in Noirmont pond (B).<br />
sis. Frog tadpoles, taken from captive<br />
breeding sites, were introduced last year<br />
for <strong>the</strong> first time (Racca 2000) and <strong>the</strong>n<br />
again this year.<br />
Population status and reproductive success<br />
<strong>of</strong> <strong>the</strong> Agile Frog in Jersey<br />
In order to produce an effective conservation<br />
programme for <strong>the</strong> agile frog in<br />
Jersey, it is necessary to investigate <strong>the</strong><br />
threats to this species and its ecology and<br />
behaviour. For this reason, on February<br />
2001, I started a three year study on <strong>the</strong><br />
ecology and conservation <strong>of</strong> Rana dalmatina<br />
in Jersey. The objectives <strong>of</strong> <strong>the</strong><br />
study are (1) to estimate <strong>the</strong> number <strong>of</strong><br />
adult frogs through a mark - recapture<br />
programme; (2) monitor hatching success<br />
and recruitment; (3) study interactions<br />
with o<strong>the</strong>r species present in <strong>the</strong> pond<br />
(toads, newts, invertebrates, and grass<br />
snakes) and in <strong>the</strong> surrounding area<br />
(grass snakes, small mammals, and birds);<br />
(4) determine <strong>the</strong> frog's overwintering<br />
preferences; (5) monitor <strong>the</strong> captive<br />
breeding programme and carry out an<br />
investigation <strong>of</strong> potential reintroduction<br />
sites; (6) develop optimal conditions for<br />
captive rearing <strong>of</strong> tadpoles and consider a<br />
health monitoring protocol prior to<br />
release <strong>of</strong> captive raised frogs; (7) compare<br />
<strong>the</strong> habitat requirements <strong>of</strong> <strong>the</strong> agile<br />
frog in Jersey and in nor<strong>the</strong>rn France.<br />
These first, preliminary results were<br />
obtained from <strong>the</strong> data collected during<br />
this year's fieldwork season, from<br />
February to July 2001. Surveys were carried<br />
out every night between 22.00-<br />
24.00hrs during <strong>the</strong> breeding season (mid<br />
January to mid March). In total, 10 male<br />
frogs were caught and PIT-tagged<br />
(Camper & Dixon 1988). No females<br />
were caught. However, from <strong>the</strong> number<br />
<strong>of</strong> egg clumps found in <strong>the</strong> breeding<br />
Figure 4. Estimated mortality <strong>of</strong> eggs during embryonic phase, in <strong>the</strong> wild (left) and in<br />
captivity (right). North Slack, South Slack and Sump are <strong>the</strong> water bodies where egg<br />
clumps were found at Ouaisne' (left) . Site A and site B are captive breeding sites; <strong>the</strong><br />
first is a private garden pond and <strong>the</strong> latter is an enclosure at <strong>the</strong> Jersey Zoo (right).<br />
1 111 ULLi<br />
North Slack South Slack<br />
• % hatched eggs D% d*ad oggs at hatching
146 Biota 3/1-2,2002 RACCA<br />
Table 1. Summary <strong>of</strong> development <strong>of</strong> wild spawn and tadpole.<br />
Number <strong>of</strong> clumps<br />
Estimated number <strong>of</strong> eggs<br />
First clump laid<br />
Minimum embryonic developmental time (days)<br />
First transformation<br />
Minimum tadpole developmental time (days)<br />
Total number <strong>of</strong> froglets caught<br />
ponds (n = 7), it can be assumed that at<br />
least this number <strong>of</strong> breeding females this<br />
year visited <strong>the</strong> site. This year, only two<br />
clumps were laid in captivity. The highest<br />
egg mortality during <strong>the</strong> embryonic<br />
phase was 40% in <strong>the</strong> wild and 50% in<br />
captivity (Figure 4). The main predators<br />
<strong>of</strong> <strong>the</strong> tadpole phase in <strong>the</strong> wild were<br />
aquatic macro-invertebrates such as<br />
Water Boatmen Notonecta glauca, Water<br />
Spiders Agryroneta aquatica, Great<br />
Diving Beetles Dytiscus marginalis, Silver<br />
Water Beetles Hydriphilus piceus, and<br />
dragonfly nymphs Odonata. These data<br />
were collected by doing monthly standardised<br />
netting sessions in <strong>the</strong> water<br />
bodies where <strong>the</strong> tadpoles were found.<br />
Palmate newts were also caught in <strong>the</strong><br />
ponds, but in very small numbers. Only<br />
three sightings <strong>of</strong> grass snakes were<br />
noted during <strong>the</strong> whole period <strong>of</strong> fieldwork.<br />
Drift fences and pitfall traps were<br />
used in order to catch newly metamorphosed<br />
frogs. Once caught, <strong>the</strong>y were<br />
marked with waterpro<strong>of</strong> ink scratched<br />
with a needle on <strong>the</strong> skin <strong>of</strong> one <strong>of</strong> <strong>the</strong>ir<br />
back legs. Sixty-eight froglets were<br />
caught and most <strong>of</strong> <strong>the</strong>m were recaptured<br />
at least once.<br />
Table 1 summarises <strong>the</strong> data collected<br />
about wild spawn clump development<br />
during this year's fieldwork.<br />
7<br />
2950<br />
23/2/01<br />
27<br />
15/6/01<br />
66<br />
68<br />
Conclusions<br />
A Species Action Plan for <strong>the</strong> agile frog<br />
has just been published in Jersey. It was<br />
written and published by <strong>the</strong> Jersey Agile<br />
Frog Group and its main objectives can be<br />
summarised as follows:<br />
1) To ensure that all existing natural,<br />
introduction and reintroduction sites are<br />
actively protected.<br />
2) To increase <strong>the</strong> number <strong>of</strong> sites<br />
through introductions / reintroductions.<br />
3) To maintain a self-sustaining captive<br />
population <strong>of</strong> frogs at a minimum <strong>of</strong><br />
three locations.<br />
4) To fur<strong>the</strong>r investigate <strong>the</strong> threats to<br />
and ecology and behaviour <strong>of</strong> <strong>the</strong> agile<br />
frog in Jersey.<br />
5) To increase <strong>the</strong> conservation pr<strong>of</strong>ile<br />
and level <strong>of</strong> awareness <strong>of</strong> <strong>the</strong> agile frog's<br />
plight in Jersey.<br />
The situation <strong>of</strong> <strong>the</strong> agile frog in Jersey is<br />
certainly worrying. On <strong>the</strong> o<strong>the</strong>r hand, an<br />
effort to try and change this situation is<br />
being made, with <strong>the</strong> publication <strong>of</strong> <strong>the</strong><br />
Species Action Plan and <strong>the</strong> involvement<br />
<strong>of</strong> <strong>the</strong> ESU, <strong>the</strong> Durrell Institute <strong>of</strong><br />
Conservation and Ecology (DICE), <strong>the</strong><br />
Durrell Wildlife Conservation Trust, <strong>the</strong><br />
Zoology Section <strong>of</strong> <strong>the</strong> Societe Jersiaise<br />
and, in <strong>the</strong> near future, with financial help<br />
from local businesses.
RACCA Biota 3/i-a, 2002 147<br />
Acknowledgements<br />
I would like to thank my supervisor Dr. Richard Griffiths for his constant help. I am also<br />
grateful to all <strong>the</strong> members <strong>of</strong> <strong>the</strong> Agile Frog Group for <strong>the</strong>ir enthusiastic involvement<br />
in this project. A grant from <strong>the</strong> Jersey Ecology Trust Fund and contributions from <strong>the</strong><br />
firm Mourant de Feu & Jeune and <strong>the</strong> Environmental Services Unit (States <strong>of</strong> Jersey<br />
Planning & Environment Committee) funded this year's PhD study.<br />
REFERENCES<br />
AGILE FROG GROUP 2000: Species Action Plan: <strong>the</strong> agile frog (Rana dalmatina) in Jersey.<br />
Environmental Services Unit, Jersey, 26 pp.<br />
CAMPER, J.D & DIXON, J.R 1988: Evaluation <strong>of</strong> a microchip marking system for amphibians<br />
and reptiles. Texas Parks and Wildlife Department, Research Publications 7100-<br />
159. Austin, Texas.<br />
CAUGHLEY, G. 1994: Directions in conservation biology. Journal <strong>of</strong> Animal Ecology, 63 (2):<br />
215-244.<br />
GROSSENBACHER, K. Rana dalmatina. In: Case, J.R, Cabela, A., Crnobrnja-lsailovic, J.,<br />
Dolmen, D., Grossenbacher, K., Haffner, P., Lescure, J., Martens, H., Martinez<br />
Rica, J.P., Maurin, H., Oliveira, M.E., S<strong>of</strong>ianidou, T.S., Veith, M. & Zuiderwijk, A.<br />
(eds.). 1997: Atlas <strong>of</strong> Amphibians and Reptiles in Europe. SHE & SPN-MNHN,<br />
Paris.<br />
GIBSON, R. & FREEMAN, M. 1997: Conservation at home; recovery programme for <strong>the</strong><br />
agile frog Rana dalmatina in Jersey. Dodo Journal <strong>of</strong> Wildlife Preservation Trust<br />
33:91-104.<br />
Jersey Legal Information Board 2001: http://www.jerseylegalinfo.je, accessed July 2001.<br />
RACCA, L. 2000: Agile frog in Jersey: a field study (March-June 2000). Environmental<br />
Services Unit, States <strong>of</strong> Jersey Planning and Environment Committee (unpublished).<br />
States <strong>of</strong> Jersey 2001: http://www.gov.je, accessed July 2001.<br />
TONGE, S. 1986: The herpet<strong>of</strong>auna in Jersey. British Herpetological Society Bulletin 17: 18-<br />
19.
SALVIDIO, ALARIO, PASTORINO & FERRETTI Biota 3/1-2, 2002 149<br />
Seasonal activity and abundance<br />
<strong>of</strong> Speleomantes ambrosii in cave<br />
habitats<br />
Sebastiano SALVIDIO1f Gabriele ALARIO1,<br />
Mauro Valeric PASTORINO2 &<br />
Mirko FERRETTI3<br />
1DIP.TE.RIS-Universita di Geneva, Corso Europa 26, 1-16132 Geneva, Italia<br />
E-mail: salvidio@dipteris.unige.it<br />
2Gruppo Speleologico Ligure "A. Issel" - Busalla (GE), Italia<br />
3Gruppo Speleologico Lunense, Sarzana (SP), Italia<br />
Abstract<br />
In this study, seasonal activity, population structure and abundance <strong>of</strong> three S. ambrosii<br />
populations were studied in three natural caves in <strong>the</strong> Province <strong>of</strong> La Spezia, NW Italy.<br />
The annual pattern <strong>of</strong> activity showed two peaks, one in spring and <strong>the</strong> o<strong>the</strong>r in<br />
autumn. The demographic structure was analysed on <strong>the</strong> basis <strong>of</strong> body-size frequency<br />
distributions, and was composed <strong>of</strong> three well separated body-size groups. The smallest<br />
component corresponded to immatures in <strong>the</strong>ir first year <strong>of</strong> life, <strong>the</strong> intermediate<br />
one to immatures aged two, and <strong>the</strong> largest comprised subadults and adults aged three<br />
years or more. Temporary removal experiments were used to estimate population abundance.<br />
Results showed that S. ambrosii populations had relatively high probability <strong>of</strong><br />
capture, ranging from 0.51 to 0.88. These results are in good accordance with previous<br />
data obtained on S. strinatii living inside an artificial cavity. Thus, cave salamanders that<br />
concentrate inside underground habitats are highly exposed to disturbance and collection.<br />
Key words: Cave habitat, population structure, Speleomantes ambrosii, removal<br />
method.<br />
Received 3 September 2001; accepted 20 February 2002
150 Biota 3/1-2,2002 SALVIDIO, ALARIO, PASTORINO & FERRETTI<br />
INTRODUCTION<br />
The cave salamander Speleomantes<br />
ambrosii (Lanza 1955) is endemic to a<br />
small karstic area in Eastern Liguria and<br />
North-western Tuscany. It displays a wide<br />
altitudinal distribution, from 30 m a.s.l. to<br />
about 1700 m in <strong>the</strong> Apuan Alps, and it is<br />
found in extremely diverse habitats from<br />
coniferous forests and maquis to rocky<br />
areas, and it usually occurs in <strong>the</strong> leaf litter,<br />
under stones along streams, and in<br />
underground habitats (Lanza et al. 1995,<br />
Lanza, 1999). The species is listed in<br />
Annex II <strong>of</strong> <strong>the</strong> "Habitat Directive"<br />
92/43/ECC and is also protected by<br />
national (DPR 357/97) and regional legislation<br />
(L.R. Liguria 4/92 and L.R.<br />
Toscana 56/00). However, little is known<br />
<strong>of</strong> <strong>the</strong> biology and ecology <strong>of</strong> this<br />
endemic species. Forti et al. (1997) and<br />
Cimmaruta et al. (1999) studied <strong>the</strong> distribution<br />
<strong>of</strong> S. ambrosiiand S. strinatii'm a<br />
parapatric area where <strong>the</strong>se species live in<br />
close contact. In this area, S. ambrosii<br />
seems confined to <strong>the</strong> more arid zones, in<br />
contrast to <strong>the</strong> more competitive S. strinatii<br />
(Cimmaruta et al. 1999).<br />
The aim <strong>of</strong> this study was to collect basic<br />
data on <strong>the</strong> seasonal activity, abundance,<br />
and population structure <strong>of</strong> S. ambrosii<br />
living in underground habitats.<br />
MATERIAL AND METHODS<br />
Activity and abundance <strong>of</strong> S. ambrosii<br />
were studied in three natural karstic caves<br />
in <strong>the</strong> Province <strong>of</strong> La Spezia, Eastern<br />
Liguria: Grotta del Papero (near Ricco del<br />
Golfo) at 205 m a.sl, Grotta Lunga di S.<br />
Antonio (near Pignone) at 230 m a.s.l.,<br />
and Grotta di Cassana (near Cassana) at<br />
190 m a.s.l. Salamanders were surveyed<br />
monthly, from September 2000 to August<br />
2001 in Grotta del Papero and Grotta<br />
Lunga di S. Antonio, while in Grotta di<br />
Cassana surveys were conducted from<br />
April to August 2001.<br />
Animals active on <strong>the</strong> cave walls were<br />
spotted with <strong>the</strong> aid <strong>of</strong> a headlamp and<br />
counted without fur<strong>the</strong>r disturbance.<br />
Population abundance was estimated by<br />
temporary removal methods (Bruce<br />
1995, Salvidio 1998, 2001). In temporary<br />
removal experiments, all animals captured<br />
are physically removed from <strong>the</strong><br />
population, kept in a holding area, and<br />
relocated when <strong>the</strong> study is completed.<br />
When basic assumptions are met, (e.g.,<br />
population closure, equal sampling effort,<br />
equal catchability, and effective reduction<br />
<strong>of</strong> <strong>the</strong> population after each search), <strong>the</strong><br />
statistics calculated from removal data are<br />
reliable. Salamanders were measured to<br />
<strong>the</strong> nearest millimetre, from <strong>the</strong> tip <strong>of</strong> <strong>the</strong><br />
snout to <strong>the</strong> posterior part <strong>of</strong> <strong>the</strong> cloaca<br />
(SVL). These SVL measurements generated<br />
polymodal frequency distribution histograms<br />
that were analysed with <strong>the</strong><br />
FAO-ICLARM Stock Assessment Tools<br />
(FiSAT) computer programme (Gayanilo<br />
et al. 1996) which enables <strong>the</strong> decomposition<br />
<strong>of</strong> mixed length-frequency distributions<br />
into <strong>the</strong>ir Gaussian components by<br />
means <strong>of</strong> Bhattacharya's (1967) log-differences<br />
method. Abundance was estimated<br />
by means <strong>of</strong> <strong>the</strong> programme CAP-<br />
TURE routine Mbh (White et al. 1982).<br />
This s<strong>of</strong>tware gives <strong>the</strong> population estimate<br />
Ni, its standard error (S.E.), and, if at<br />
least three removal samplings are used, it<br />
allows a test for homogeneity <strong>of</strong> capture<br />
probabilities with a x2 goodness <strong>of</strong> fit test.<br />
During this study no direct mortality <strong>of</strong><br />
<strong>the</strong> removed salamanders occurred, and<br />
at <strong>the</strong> end <strong>of</strong> <strong>the</strong> third removal sampling<br />
all salamanders were released at <strong>the</strong>ir<br />
capture sites.<br />
RESULTS<br />
Seasonal activity<br />
The activity patterns in <strong>the</strong> three caves<br />
were similar (Figure 1), and annual trends<br />
were highly correlated in Grotta del<br />
Papero and Grotta di S. Antonio<br />
(Spearman rank correlation coefficient: rs<br />
= 0.90, n =12; P < 0.01), suggesting that<br />
similar extrinsic (e.g., climate, food abun-
SALVIDIO, ALARIO, PASTORINO & FERRETTI Biota 3/1-2,2002 151<br />
Figure 1. Seasonal pattern <strong>of</strong> underground activity in three cave populations <strong>of</strong><br />
Speleomantes ambrosii.<br />
Number <strong>of</strong> individuals<br />
Sep Oct Nov Dec Jan Feb Mar Apr May Jun Jul Aug<br />
dance) and/or intrinsic (e.g., physiological<br />
cycle) factors were acting in all sites<br />
simultaneously.<br />
Abundance <strong>of</strong> cave populations<br />
The statistics resulting from <strong>the</strong> temporal<br />
removal experiments are given in Table 1.<br />
Capture probabilities were always higher<br />
than 0.20, a value considered adequate in<br />
estimating population abundance in<br />
removal experiments (White et al. 1982).<br />
These results indicate that <strong>the</strong> assumption<br />
<strong>of</strong> closure were met. However, <strong>the</strong> numbers<br />
<strong>of</strong> removed salamanders from both<br />
Grotta del Papero and Grotta Lunga di S.<br />
Antonio were too low to represent well<br />
structured populations. On <strong>the</strong> contrary,<br />
70 specimens were captured inside<br />
Papero<br />
S. Antonio<br />
Cassana<br />
Grotta di Cassana, suggesting that this<br />
latter cave was a more favourable habitat.<br />
Population structure<br />
All <strong>the</strong> salamanders captured in July 2001<br />
removal experiments (n = 106) were<br />
pooled to obtain an adequate sample to<br />
be analysed. Body-size measurements<br />
generated a polymodal frequency distribution<br />
histogram that was success<strong>full</strong>y<br />
decomposed into three separated gaussian<br />
components (Figure 2). The two<br />
smallest body-size groups (mean SVL =<br />
25.99, S.D. 2.67, and mean SVL = 41.11,<br />
S.D. 4.13) were composed <strong>of</strong> immature<br />
salamanders, while <strong>the</strong> largest one (mean<br />
SVL = 60.28, S.D. 5.44) was a mixture <strong>of</strong><br />
Table 1. Speleomantes ambrosii removal data and population size estimates. Ni =<br />
Number estimated; S.E. = standard error; (Chi-square test for homogeneity <strong>of</strong> capture<br />
probabilities (P).<br />
Year 2000<br />
Grotta di S. Antonio<br />
Grotta del Papero<br />
Year 2001<br />
Grotta di S. Antonio<br />
Grotta del Papero<br />
Grotta di Cassana<br />
1st capture<br />
June 25<br />
19<br />
24<br />
1st capture<br />
July?<br />
10<br />
13<br />
42<br />
2M capture<br />
June 27<br />
3<br />
3<br />
2 capture<br />
July 9<br />
2<br />
8<br />
15<br />
3"" capture<br />
June 29<br />
0 1<br />
31 capture<br />
July 11<br />
2<br />
1<br />
13<br />
Ni<br />
22<br />
28<br />
14<br />
22<br />
79<br />
S.E.<br />
0.21<br />
0.34<br />
0.82<br />
1.25<br />
6.27<br />
X2 P Capture<br />
probability<br />
0.52 >0.05 0.88<br />
0.58 >0.05 0.85<br />
2.10<br />
2.77<br />
2.35<br />
>0.05<br />
>0.05<br />
>0.05<br />
0.70<br />
0.67<br />
0.51
152 3/1-2, 2OO2 SALVIDIO, ALARIO, PASTORINO & FERRETTI<br />
Figure 2. Speleomantes ambrosii population structure. The curves separating each<br />
Gaussian component were calculated by <strong>the</strong> FiSAT s<strong>of</strong>tware (see text).<br />
20 24 28 32 36 40<br />
subadults (i.e.r large immatures) and<br />
adults (i.e., gravid females and males with<br />
mental glands). If females reach sexual<br />
maturity at about 58 mm in SVL, as in S.<br />
strinatii (Salvidio 1993), <strong>the</strong>n <strong>the</strong> sex ratio<br />
<strong>of</strong> <strong>the</strong> adult population was not different<br />
from 1 (17 males/9 females; Chi-square =<br />
2.46, P > 0.05).<br />
DISCUSSION<br />
In <strong>the</strong> study caves, seasonal activity was<br />
clearly bimodal, peaking in summer and<br />
autumn, and during winter few active<br />
salamanders were observed on <strong>the</strong> cave<br />
walls. Overall, this pattern <strong>of</strong> activity was<br />
similar to that <strong>of</strong> a S. strinatii population<br />
inhabiting a rock-face habitat (Salvidio<br />
1993). This activity pattern was in agreement<br />
between sites, suggesting that <strong>the</strong><br />
same extrinsic or intrinsic factors were<br />
acting in all caves, which were in fact at<br />
similar altitudes and <strong>the</strong>refore exposed to<br />
similar climatic conditions.<br />
The demographic structure was composed<br />
<strong>of</strong> three Gaussian components,<br />
N-106<br />
48 52 53 60 64 68<br />
and two cohorts <strong>of</strong> immatures in <strong>the</strong>ir<br />
first and second year <strong>of</strong> life were<br />
observed. The growth increment<br />
between <strong>the</strong>se two components was<br />
about 15 mm, a higher value in relation<br />
to that reported for a rock-face population<br />
<strong>of</strong> S. strinatii, in which juvenile<br />
growth was about 10-13 mm in <strong>the</strong> first<br />
year (Salvidio 1998). Within <strong>the</strong> largest<br />
body size group, it was not possible to<br />
separate o<strong>the</strong>r age classes, as growth rate<br />
decreases in larger (and older) individuals<br />
having reached sexual maturity.<br />
In <strong>the</strong> caves Grotta del Papero and Grotta<br />
Lunga di S. Antonio, few salamanders<br />
were removed from <strong>the</strong> cave walls, suggesting<br />
that <strong>the</strong>se sites hosted only a limited<br />
proportion <strong>of</strong> salamanders living in<br />
<strong>the</strong> surrounding karst system. On <strong>the</strong><br />
contrary, in <strong>the</strong> cave Grotta di Cassana,<br />
<strong>the</strong> abundance <strong>of</strong> salamanders was adequate<br />
to obtain effective estimates <strong>of</strong><br />
population abundance. Overall, 70 salamanders<br />
were removed and capture<br />
probabilities were relatively high (i.e.,
SALVIDIO, ALARIO, PASTORINO & FERRETTI BJOta 3/1-2, 2002 153<br />
0.51) suggesting that about half <strong>of</strong> <strong>the</strong><br />
population was exposed to capture during<br />
each capture session. This value was<br />
similar to that obtained for S. strinatii living<br />
inside an artificial tunnel (mean value<br />
0.639; Salvidio 2001). Thus, Grotta di<br />
Cassana was a favourable habitat for<br />
salamanders that were highly exposed to<br />
disturbance and collection, at least during<br />
<strong>the</strong>ir underground peaks <strong>of</strong> activity.<br />
These results have implications for conservation,<br />
as those caves hosting large<br />
numbers <strong>of</strong> salamanders should be closed<br />
or monitored to avoid disturbance and<br />
overcollection <strong>of</strong> <strong>the</strong> endemic S. ambrosii.<br />
Acknowledgements<br />
Capture permits were obtained from <strong>the</strong> Italian A/linistero deH'Ambiente - Servizio<br />
Conservazione della Natura (prat. n. SCN/2D/98/14345 and SCN/2D/2001/378). We<br />
thank Edoardo Razzetti for his field help during <strong>the</strong> July 2000 sampling.<br />
REFERENCES<br />
BRUCE, R. C. 1995: The use <strong>of</strong> temporary removal sampling in a study <strong>of</strong> population dynamics<br />
<strong>of</strong> <strong>the</strong> salamander Desmognathus monticola. Australian Journal <strong>of</strong> Ecology<br />
20: 403-412.<br />
CIMMARUTA, R., FORTI, G., NASCETTI, G. & BULLINI, L. 1999: Spatial distribution and<br />
competition in two parapatric sibling species <strong>of</strong> European plethodontid salamanders.<br />
Ethology Ecology and Evolution 11: 383-398.<br />
FORTI, G., CIMMARUTA, R., NASCETT, G. & BULLINI, L. 1997: Parapatria e competizione<br />
in pletodontidi del genere Hydromantes. S.ltE Atti 18: 121-124.<br />
GAYANILO, Jr., F.C., SPARRE, P. & PAULY, D. 1996: The FAO-ICLARM Stock Assessment<br />
Tools (FiSAT) User's Guide. FAO Computerized Information Series (Fisheries), 8.<br />
Food and Agriculture Organisation <strong>of</strong> <strong>the</strong> United Nations. Rome.<br />
LANZA, B. 1999: Plethodontidae - Lungenlose Salamander. In: Grossenbacher, K. &<br />
Thiesmeier, B. (eds.). Handbuch der Reptilien und Amphibien Europas. Aula<br />
Verlag, Wiesbaden: 77-204.<br />
LANZA, B., CAPUTO, V., NASCETTI, G. & BULLINI, L. 1995: Morphological and genetic<br />
studies <strong>of</strong> <strong>the</strong> European plethodontid salamanders: taxonomic inferences (genus<br />
Hydromantes). Torino, Mus. reg. Sci. nai., Monografie 16, Torino.<br />
SALVIDIO, S. 1993: Life history <strong>of</strong> <strong>the</strong> European plethodontid salamander Speleomantes<br />
ambrosii. Herpetological Journal 3: 55-59.<br />
SALVIDIO, S. 1998: Estimating abundance and biomass <strong>of</strong> a Speleomantes strinatii<br />
(Caudata: Plethodontid) population by temporary removal sampling. Amphibia-<br />
Reptilia19: 113-124.<br />
SALVIDIO, S. 2001: Estimating terrestrial salamander abundance in different habitats: efficiency<br />
<strong>of</strong> temporary removal methods. Herpetological Review 32: 21-24.<br />
SALVIDIO, S. LATTES, A., TAAVANO, M., MELODIA, F. & PASTORINO, M.V. 1994: Ecology<br />
<strong>of</strong> a Speleomantes ambrosii population inhabiting an artificial tunnel. Amphibia-<br />
Reptilia 15: 35-45.<br />
WHITE, G.C., ANDERSON, D.R., BURNHAM, K.P. & OTIS, D.L. 1982: Capture-recapture<br />
and Removal Methods for Sampling Closed Populations. Los Alamos National<br />
Laboratory 8787 NERP, Los Alamos, New Mexico. 235 pp.
SCALI & GENTILLI Biota 3/1-2.2002 155<br />
A comparison <strong>of</strong> main heathlands<br />
in nor<strong>the</strong>rn Italy and <strong>the</strong>ir<br />
importance for amphibian<br />
populations<br />
Stefano SCALP & Augusto GENTILLI2<br />
'Museo Civico di Storia Naturale di Milano, C.so Venezia 55 - 1-20121 Milano<br />
E-mail: scaliste@iol.it<br />
2Dipartimento di Biologia Animale, Universita di Pavia, P.zza Botta 9-10, 1-27100 Pavia<br />
E-mail: augusto.gentilli@unimib.it<br />
Abstract<br />
The importance <strong>of</strong> heathlands as habitat for reptiles has been stressed by many authors.<br />
Few studies about <strong>the</strong>ir importance for amphibians, on <strong>the</strong> o<strong>the</strong>r hand, are available<br />
and <strong>the</strong> significance <strong>of</strong> <strong>the</strong>se habitats is probably underestimated. Heathlands are considered<br />
priority natural habitats by <strong>the</strong> European Union in <strong>the</strong> Habitats Directive and so<br />
<strong>the</strong>ir ecology is particularly important to understand.<br />
The authors conducted some herpetological research in <strong>the</strong> main heathlands in <strong>the</strong> Po<br />
Plain (Nor<strong>the</strong>rn Italy) and combined new data with bibliographic information. Six<br />
heathland areas were investigated in Piedmont and Lombardy, and three to seven<br />
amphibian species were found; <strong>the</strong> complete list <strong>of</strong> taxa recorded in all <strong>the</strong> heathlands<br />
11 is <strong>the</strong> following: Triturus carnifex, T. vulgaris meridionalis, Pelobates fuscus insubricus,<br />
* f Bufo bufo, B. viridis, Hyla intermedia, Rana dalmatina, R. latastei, R. temporaria, R. synklepton<br />
esculenta.<br />
Some information about water presence and permanence, vegetation structure and<br />
coverage, soil type and distance from <strong>the</strong> Alps is also presented. Data were analysed<br />
using multivariate techniques. Two areas were very different from <strong>the</strong> o<strong>the</strong>rs: <strong>the</strong> first<br />
one is a deteriorating heathland, devoid <strong>of</strong> woods and permanent water, with a low<br />
amphibian species diversity; <strong>the</strong> second one is <strong>the</strong> sou<strong>the</strong>rnmost area and it is characterised<br />
by <strong>the</strong> presence <strong>of</strong> many scattered trees. The o<strong>the</strong>r four zones were quite similar,<br />
even if some differences could be observed in amphibian species composition and<br />
habitat structure.<br />
Key words: Amphibians, conservation, heathland, Nor<strong>the</strong>rn Italy<br />
Received 2 October; accepted 8 February 2002
156 Biota 3 1-5 SCALI & GENTILLI<br />
INTRODUCTION<br />
Heathlands are considered priority natural<br />
habitats by <strong>the</strong> European Union in <strong>the</strong><br />
Habitat Directive 92/43/CEE, Enclosure<br />
1, CORINE habitat code 31.2 because<br />
<strong>the</strong>y have strongly declined throughout<br />
Europe (Beebee 1996). Therefore, knowledge<br />
<strong>of</strong> <strong>the</strong>ir fauna is particularly important<br />
for conservation projects.<br />
The importance <strong>of</strong> heathlands as habitat<br />
for reptiles has been stressed by many<br />
authors (Spellerberg 1989, Stumpel<br />
1992, Scali 1995, Zuffi 1987a, b). Few<br />
studies about <strong>the</strong>ir importance for<br />
amphibians, on <strong>the</strong> o<strong>the</strong>r hand, are available<br />
(Scali 1993, Beebee 1996, Denton &<br />
Beebee 1996) and <strong>the</strong> significance <strong>of</strong><br />
<strong>the</strong>se habitats is probably underestimated.<br />
Many amphibian species that live in<br />
heathlands are considered very important<br />
taxa in <strong>the</strong> Habitats Directive: Triturus<br />
earn if ex, Pelo bates fuscus insubricus,<br />
Bufo calamita, Bufo viridis, Hyla arborea,<br />
Hyla intermedia, Rana arvalis, Rana dalmatina,<br />
Rana latastei and Rana lessonae.<br />
MATERIALS AND METHODS<br />
The authors conducted some herpetological<br />
research in <strong>the</strong> main heathlands <strong>of</strong><br />
<strong>the</strong> western Po Plain (Nor<strong>the</strong>rn Italy) during<br />
<strong>the</strong> last ten years, and <strong>the</strong>y combined<br />
new data with bibliographic information<br />
(Pozzi 1980). Six heathland areas were<br />
investigated in Piedmont and Lombardy:<br />
1. Piano Rosa, 2. Rovasenda, 3. Lonate<br />
Pozzolo, 4. Mombello, 5. Ca' del Re, and<br />
6. Salvadorino (Figure 1).<br />
We prepared a matrix using <strong>the</strong> following<br />
variables: area, presence <strong>of</strong> <strong>the</strong> different<br />
taxa, total number <strong>of</strong> taxa, altitude, distance<br />
from <strong>the</strong> Alps, presence and number<br />
<strong>of</strong> permanent and temporary ponds,<br />
presence <strong>of</strong> some plant species (Pinus<br />
sylvestris, Betu/a pendu/a, Populus sp.,<br />
Quercus sp., Pteridium aquilinum), presence<br />
<strong>of</strong> woods in or around <strong>the</strong> area, and<br />
type <strong>of</strong> soil (clay or gravel). A summary <strong>of</strong><br />
all <strong>the</strong> variables and <strong>the</strong>ir abbreviations<br />
(in paren<strong>the</strong>ses) is as follows: i) heathland<br />
ID (Area), ii) distance from <strong>the</strong> Alps<br />
(Alps), iii) number <strong>of</strong> permanent ponds<br />
Figure 1. Location <strong>of</strong> <strong>the</strong> study areas in Nor<strong>the</strong>rn Italy. Heathlands are assigned by same<br />
numbers as in tables 1 and 2.<br />
__ Chattllon<br />
VALLFD.MlSTft<br />
TraverseHi. "*ji<br />
'Atogna Campo<br />
y<br />
- Breia \ Angara •.<br />
'• '"' Gozzano'1<br />
D Vv V^Sesto Sesto Catencte<br />
A8argcise-;ia , . '-*-.--<br />
.c\*ne<br />
v-.^<br />
DCantV<br />
n SonmaLambaVdO: ' Tr8dale Mar^nDC«ne<br />
"Cogsfale . 1 V: ...•., ° >JGallarete<br />
; _0urttengo<br />
" i -;/ * ~ *i' Pralungo.<br />
Peninefiflo<br />
Gattinaran<br />
•;<br />
Ghemme<br />
n .<br />
I1 '-ffusK Arsizio, .'-<br />
N :;,, i '<br />
°0lerao .nhonado'Lognalio .<br />
^_<br />
3_<br />
09ena90<br />
^Bislla _ --UJsona* -suano caston!)Primb ' Nerviam,<br />
:ambgijano •* ... :•. . '1-1^1 \n Jnveruno ° °<br />
% "a'vrea<br />
J^eroJo Canevese<br />
Socca Cana»t5« ; ^CaluSD<br />
.<br />
FelBto"<br />
J<strong>of</strong>razjo Vterron* ^h4maleiata " .Undwna<br />
.CV.L.biano<br />
" ° a<br />
^S^t^N ^ r , r-Cassolnovo<br />
TrOnzano VerceSese Bbrgo.Vercelli '•; °<br />
D - - ..c ..-'•-".«!. . "7 vigevano
SCALI & GENTILLI Biota 3/i-a, 2002 157<br />
Table 1. Habitat characteristics <strong>of</strong> <strong>the</strong> six analysed heathlands. Presence <strong>of</strong> plant species<br />
is coded as follows: 0=absent, 1=present; soil is coded as 1=clay, 2=gravel.<br />
Area<br />
1<br />
2<br />
3<br />
4<br />
5<br />
6<br />
Alps<br />
(Km)<br />
5<br />
15<br />
30<br />
25<br />
25<br />
65<br />
Permsites<br />
(N)<br />
1<br />
1<br />
0<br />
2<br />
1<br />
1<br />
Tempsites<br />
(N)<br />
2<br />
1<br />
6<br />
4<br />
3<br />
0<br />
(Permsites), iv) Number <strong>of</strong> temporary<br />
ponds (Tempsites), v) presence <strong>of</strong> permanent<br />
water (Permwater), vi) presence <strong>of</strong><br />
temporary water (Tempwater), vii) presence<br />
<strong>of</strong> Pinus sylvestris (Pinus), viii) presence<br />
<strong>of</strong> Betula pendula (Betula), ix) presence<br />
<strong>of</strong> Populus sp. (Populus), x) presence<br />
<strong>of</strong> Quercus sp. (Quercus), xi) presence<br />
<strong>of</strong> Pteridium aquilinum (Pteridium),<br />
xii) percentage <strong>of</strong> woods in <strong>the</strong> area<br />
(Intwoods), xiii) percentage <strong>of</strong> woods<br />
around <strong>the</strong> area (Extwoods), xiv) type <strong>of</strong><br />
soil (Soil). The six areas were coded as<br />
wet or dry heathlands: <strong>the</strong> former ones<br />
have clayey and waterpro<strong>of</strong> soils, with<br />
hygrophilous vegetation, while <strong>the</strong> latter<br />
ones have gravelly and permeable soils<br />
and plants typical <strong>of</strong> dry areas. Two types<br />
<strong>of</strong> degradation can be found in <strong>the</strong>se<br />
heathlands: A) a lack <strong>of</strong> management<br />
that leads to a succession through to<br />
woodland mainly composed <strong>of</strong> exotic<br />
species, such as Robinia pseudoacada<br />
and Prunus serotina, B) a heavy human<br />
disturbance deriving from many kinds <strong>of</strong><br />
activities (i.e. road traffic, pollution, water<br />
drainage, buildings, etc.).<br />
The six studied areas (Figure 1) are<br />
described briefly below and in Table 1:<br />
1. Piano Rosa (300 m a.s.l.): it is a medium-sized,<br />
well preserved, wet heathland<br />
near <strong>the</strong> Alps, with sparse woods and<br />
large open areas. It is surrounded by<br />
woods and agricultural lands. A small<br />
number <strong>of</strong> permanent and temporary<br />
ponds are present.<br />
2. Rovasenda (220 m a.s.l.): it is a large,<br />
well preserved, wet heathland. Human<br />
Permwater Tempwater Pinus<br />
(N) (N)<br />
1<br />
1<br />
0<br />
1<br />
0<br />
0<br />
0<br />
1<br />
1 ( ) 0<br />
Betula<br />
1<br />
1<br />
0<br />
1<br />
1<br />
1<br />
Populus<br />
1<br />
0<br />
0<br />
1<br />
1<br />
1<br />
Quercus<br />
disturbance is low and many different<br />
microhabitats are present. A stream and<br />
some permanent and temporary ponds<br />
are present.<br />
3. Lonate Pozzolo (220 m a.s.l.): it is a<br />
large, deteriorating, wet area with small,<br />
well preserved portions. Human disturbance<br />
is quite heavy, due to military<br />
activities and road construction; a progressive<br />
growth <strong>of</strong> wood is also present.<br />
A thin woodland belt is present around<br />
<strong>the</strong> area. Only a small number <strong>of</strong> temporary<br />
ponds are present.<br />
4. Mombello (210 m a.s.l.): it is a medium-sized,<br />
deteriorating, wet heathland<br />
once used as a clay quarry. Human disturbance<br />
is quite low and caused by quarry<br />
activities on <strong>the</strong> borders and wood<br />
growth. The area is characterised by <strong>the</strong><br />
presence <strong>of</strong> many little rises covered by<br />
shrubs and woods. Two large, permanent<br />
and many temporary ponds are present.<br />
5. Ca' del Re (210 m a.s.l.): it is a medium-sized,<br />
well preserved, wet heathland<br />
located near <strong>the</strong> previous area. Human<br />
disturbance is low. Some sparse woods<br />
are present in <strong>the</strong> area and a deteriorating<br />
woodland belt surrounds <strong>the</strong> heathland.<br />
A large permanent pond and some temporary<br />
ponds are present.<br />
6. Salvadorino (70 m a.s.l.): it is a very<br />
small, well preserved, dry area with some<br />
isolated trees and shrubs. It is surrounded<br />
by typical plain woods and channels.<br />
Only one small permanent pond is present.<br />
Data were analysed using two multivariate<br />
methods (cluster analysis with <strong>the</strong><br />
0<br />
1<br />
0<br />
0<br />
0<br />
I
158 Biota 3/1-2,2002 SCALI & GENTILLI<br />
Table 2. Presence <strong>of</strong> <strong>the</strong> ten Amphibian species in <strong>the</strong> six areas.<br />
Area<br />
1<br />
2<br />
3<br />
4<br />
5<br />
6<br />
Tricar<br />
1<br />
1<br />
0 1<br />
1<br />
0<br />
Trivul<br />
1<br />
1<br />
0<br />
0<br />
1<br />
1<br />
Pelftis<br />
0 1<br />
0<br />
0<br />
0<br />
0<br />
Bufbuf<br />
J<br />
0<br />
0<br />
0<br />
0 1<br />
Bufvir Hylint<br />
0<br />
0 1<br />
1<br />
0<br />
0<br />
UPGMA method and correspondence<br />
analysis) with <strong>the</strong> MVSP 3.1 for Windows<br />
package. These two techniques were<br />
chosen to obtain some groups <strong>of</strong> related<br />
data; cluster analysis was used to group<br />
<strong>the</strong> heathlands in relation to habitat characteristics<br />
and presence <strong>of</strong> different<br />
species. Correspondence analysis allowed<br />
us to group different variables and<br />
species to understand <strong>the</strong> relationships<br />
between <strong>the</strong>m.<br />
Figure 2. Dendrogram <strong>of</strong> <strong>the</strong> grouped<br />
heathlands in relation to habitat characteristics<br />
and species composition<br />
(UPGMA method).<br />
0,52 0,6 0,68 0,76 0,84 0,92 1<br />
-2<br />
-5<br />
-4<br />
-1<br />
RESULTS<br />
Three to seven amphibian species were<br />
found in <strong>the</strong> six heathlands (Table 2); <strong>the</strong><br />
complete list <strong>of</strong> taxa recorded in all <strong>the</strong><br />
heathlands is as follows: Triturus carnifex,<br />
T. vulgaris meridionalis, Pelobates fuscus<br />
insubricus, Bufo bufo, B. viridis, Hyla<br />
intermedia, Rana dalmatina, R. latastei, R.<br />
Randal<br />
1<br />
0<br />
0 1<br />
1<br />
1<br />
Ranlat<br />
0<br />
0<br />
0<br />
0<br />
1<br />
1<br />
Ranesc<br />
1<br />
1<br />
1<br />
1<br />
1<br />
1<br />
Rantem<br />
J<br />
0<br />
0<br />
0<br />
0<br />
0<br />
No. <strong>of</strong> species<br />
7<br />
5<br />
3<br />
5<br />
6<br />
6<br />
temporaria, and R. synklepton esculenta.<br />
A cluster analysis was conducted on <strong>the</strong><br />
basis <strong>of</strong> <strong>the</strong> data reported in Tables 1 and<br />
2 and <strong>the</strong> results are showed in Figure 2.<br />
Two areas (3 and 6) are well separated by<br />
a group <strong>of</strong> similar areas (1,2,4 and 5).<br />
The presence <strong>of</strong> all species was related to<br />
environmental variables using <strong>the</strong> correspondence<br />
analysis to highlight habitat<br />
preferences <strong>of</strong> amphibians in <strong>the</strong>se<br />
heathlands. The results <strong>of</strong> this analysis are<br />
reported in Figure 3. Three main clusters<br />
were highlighted: i) Bufo viridis was<br />
grouped with <strong>the</strong> presence and <strong>the</strong> number<br />
<strong>of</strong> temporary sites, ii) Rana latastei<br />
and Bufo bufo were grouped with natural<br />
woodlands internal to studied areas, iii)<br />
Triturus vulgaris, Hyla intermedia and<br />
Rana synklepton esculenta were associated<br />
with permanent sites and surrounding<br />
woodlands. The o<strong>the</strong>r species appear to<br />
be less associated with <strong>the</strong> considered<br />
variables.<br />
DISCUSSION<br />
The six heathland areas <strong>of</strong> <strong>the</strong> western Po<br />
Plain comprise good habitat for amphibians<br />
and <strong>the</strong> presence <strong>of</strong> ten species was<br />
recorded altoge<strong>the</strong>r. The minimum<br />
recorded number <strong>of</strong> species was three<br />
and <strong>the</strong> maximum was seven. It is particularly<br />
interesting that some endangered<br />
taxa - such as Pelobates fuscus insubricus<br />
and Rana latastei ~ are present. These<br />
species usually live in wood land areas,<br />
but can also colonize some well-preserved<br />
heathlands.<br />
Our analyses showed a certain similarity<br />
in environmental characteristics and
SCALI & GENTILLI )ta 3/1-2, 2OO2 159<br />
amphibian species composition between<br />
<strong>the</strong> six considered areas.<br />
Only two areas were very different from<br />
<strong>the</strong> o<strong>the</strong>rs: <strong>the</strong> first one, (3), is a deteriorating<br />
heathland, devoid <strong>of</strong> woodland<br />
and permanent water, with a low<br />
amphibian species diversity. The scarcity<br />
<strong>of</strong> amphibians in this zone confirms <strong>the</strong>ir<br />
sensitivity to habitat degradation. The<br />
second area, (6), is <strong>the</strong> sou<strong>the</strong>rnmost area<br />
and it is characterised by <strong>the</strong> presence <strong>of</strong><br />
many scattered trees; <strong>the</strong> different habitat<br />
structure probably creates some<br />
microclimatic conditions that increase<br />
amphibian diversity. The o<strong>the</strong>r four zones<br />
are quite similar, even if some differences<br />
could be observed in amphibian species<br />
composition and habitat structure.<br />
In particular, <strong>the</strong> presence <strong>of</strong> <strong>the</strong> Green<br />
Toad is correlated to temporary ponds, in<br />
accordance with <strong>the</strong> information available<br />
for this species (Nollert & Nollert<br />
1992, Scali 1995). The Common Toad<br />
and <strong>the</strong> Italian Agile Frog are present only<br />
in heathlands with internal woodlands,<br />
characterized by typical lowland trees (i.e.<br />
Quercus robur) (Borkin & Veith 1997,<br />
Grossenbacher 1997). The Smooth Newt,<br />
<strong>the</strong> Italian Treefrog and <strong>the</strong> Edible Frog<br />
need external woodlands, with large and<br />
sunny permanent ponds (Lanza 1983).<br />
Rana temporaria is only influenced by <strong>the</strong><br />
proximity to <strong>the</strong> Alps, because it usually<br />
lives in mountainous areas in Italy (Lanza<br />
1983). The Italian Spadefoot Toad was<br />
found only once by Pozzi (1980) in<br />
Rovasenda heathland; its presence should<br />
be considered occasional, because this<br />
species usually prefers areas with sandy<br />
soils (Lanza 1983). It is quite difficult to<br />
explain <strong>the</strong> habitat preferences <strong>of</strong> <strong>the</strong><br />
Italian Crested Newt; in particular, those<br />
preferences which do not appear clearly<br />
related to <strong>the</strong> variables used in <strong>the</strong> present<br />
study.<br />
Our results confirm that heathlands could<br />
be important for <strong>the</strong> preservation <strong>of</strong> some<br />
amphibian populations and for <strong>the</strong> maintenance<br />
<strong>of</strong> high amphibian diversity.<br />
Figure 3. Association between <strong>the</strong> variables calculated with correspondence analysis<br />
(Axis 1: Eigenvalue = 0.160, Percentage <strong>of</strong> variance = 74.7; Axis 2: Eigenvalue = 0.029,<br />
Percentage <strong>of</strong> variance = 13.5).<br />
CA variable scores
160 Biota 3/1-2,2002 SCALI & GENTILLI<br />
REFERENCES<br />
BEEBEE, TJ.C. 1996: Ecology and Conservation <strong>of</strong> Amphibians. Chapman & Hall, London:<br />
214 pp.<br />
BORKIN, L.J. & VEITH, M. 1997: Bufo bufo (Linnaeus, 1758). In: Case, J.P., Cabela, A.,<br />
Crnobrnja-lsailovic, J., Haffner, P., Lescure, J., Martens, H., Martinez Rica, J.P.,<br />
Maurin, H., Oliveira, T., S<strong>of</strong>ianidou, T.S., Veith, M. & Zuiderwijk, A. (eds.), Atlas<br />
<strong>of</strong> Amphibians and Reptiles in Europe. Societas Europea Herpetologica &<br />
Museum National d'Histoire Naturelle (IEGP/SPN), Paris: 118-119.<br />
DENTON, J.S. & BEEBEE, TJ.C. 1996: Habitat occupancy by juvenile natterjack toads (Bufo<br />
calamita) on grazed and ungrazed heathland. Herpetological Journal 6: 49-52.<br />
GROSSENBACHER, K. 1997: Rana latastei Boulenger, 1879. In: Case, J.P., Cabela, A.,<br />
Crnobrnja-lsailovic, J., Haffner, P., Lescure, J., Martens, H., Martinez Rica, J.P.,<br />
Maurin, H., Oliveira, T., S<strong>of</strong>ianidou, T.S., Veith, M. & Zuiderwijk, A. (eds.), Atlas<br />
<strong>of</strong> Amphibians and Reptiles in Europe. Societas Europea Herpetologica &<br />
Museum National d'Histoire Naturelle (IEGP/SPN), Paris: 146-147.<br />
LANZA, B. 1983: Anfibi e Rettili. Guide per il riconoscimento delle specie animali delle acque<br />
interne italiane. 27. Anfibi, Rettili (Amphibia, Reptilia) [Collana del progetto finalizzato<br />
"Promozione della qualita deH'ambiente". AQ/1/205]. Roma; Consiglio<br />
Nazionale delle Ricerche.<br />
NOLLERT, A. & NOLLERT, C. 1992: Die Amphibien Europas: Bestimmung - Gefahrdung -<br />
Schutz. Franckh-Kosmos, Stuttgart.<br />
POZZI, A. 1980: Anfibi e Rettili della brughiera di Rovasenda (Piemonte). AQ/1756-67.<br />
Quaderni sulla "Struttura delle Zoocenosi terrestri". C.N.R., Roma: 467-472.<br />
SCALI, S. 1993: Osservazioni su Rana latastei e Triturus vulgaris meridionals nel Parco<br />
delle Groane (Lombardia, Italia). In: Ferri, V. (ed.). Atti del Primo Convegno<br />
Italiano sulla Salvaguardia degli Anfibi, Quaderni della Civica Stazione<br />
Idrobiologica Milano 20: 109-116.<br />
SCALI, S. 1995: Amphibians and reptiles <strong>of</strong> Groane Regional Park (Lombardy, NW Italy).<br />
First census and ecological notes. In: Llorente, G.A., Montori, A., Santos, X. &<br />
Carretero, M.A. (Eds.). Scientia Herpetologica. Asociacion Herpetologica<br />
Espanola, Barcelona: 307-311.<br />
SPELLERBERG, I.F. 1989: An assessment <strong>of</strong> <strong>the</strong> importance <strong>of</strong> heathlands as habitats for reptiles.<br />
Botanical Journal <strong>of</strong>,<strong>the</strong> Linnean Society 101: 313-318.<br />
STUMPEL, A.H.P. 1992: Reptile management problems in Ne<strong>the</strong>rlands heathlands. In:<br />
Korsos, Z. & Kiss, I. (eds.), Proc. Sixth Ord. Gen. Meet. S.E.H, Budapest, 1991:<br />
421-424.<br />
ZUFFI, M. 1987a: Anfibi e Rettili del Parco lombardo della Valle del Ticino: risultati preliminari<br />
e proposte gestionali. Quaderni Civica Stazione Idrobiologica, Milano 14: 7-<br />
65.<br />
ZUFFI, M. 1987b: Su alcune stazioni di Podarcis sicula campestris (De Betta, 1857) della<br />
Lombardia occidentale (Reptilia Lacertidae). Atti della Societa italiana di Scienze<br />
natural! e del Museo civico di Storia naturale, Milano 128: 169-172.
SCALI, CORTI, GENTILLI, LUISELLI, RAZZETTI & ZUFFI Biota 3/i-a, 2002 161<br />
Continental versus Mediterranean<br />
European Whip Snake Hierophis<br />
viridiflavus: a morphometric<br />
approach<br />
Stefano SCALI1, Claudia CORTPf Augusto<br />
GENTILLI3, Luca LUISELLI4, Edoardo<br />
RAZZETTI5 & Marco A.L. ZUFFI5*<br />
'Museo Civico di Storia Naturale, c.so Venezia 55, 20121 Milano-ltaly<br />
2Dip. Biologia Animale e Genetica, University <strong>of</strong> Florence, via Romana 17, 50125<br />
Florence-Italy<br />
3Dip. Biologia Animale, University <strong>of</strong> Pavia, p.za Botta 9, 27100 Pavia, Italy<br />
"Istituto di Studi Ambientali "Demetra" (E.N.I. S.p.A.), via dei Cochi 48/B, 1-00133<br />
Rome, Italy; and F.I.Z.V., via Cleonia 30, 1-00152 Rome, Italy; and Department <strong>of</strong><br />
Biological Sciences, Rivers State University <strong>of</strong> Science and Technology, P.M.B. 5080, Port<br />
Harcourt (Rivers State)-Nigeria<br />
^Corresponding author: Museo di Storia Naturale e del Territorio, University <strong>of</strong> Pisa, via<br />
Roma 79, 1-56011 Calci (Pisa)-ltaly<br />
E-mail: marcoz@museo.unipi.it.<br />
Abstract<br />
Hierophis viridiflavus, a wide distributed European colubrid, has been subjected to taxonomic<br />
studies, with particular reference to morphological aspects <strong>of</strong> subspecific variation.<br />
O<strong>the</strong>r aspects <strong>of</strong> biology, ecology, and ethology are still anecdotal. Because <strong>of</strong> its<br />
wide distribution and extreme abundance around <strong>the</strong> Tyrrhenian and Ligurian Seas, it<br />
could be <strong>of</strong> great interest to test if any eventual geographic variation <strong>of</strong> morphological<br />
features could be related to different ecological adaptations (i.e.: dietary habits, reproductive<br />
strategies). We focused our attention on <strong>the</strong> occurence <strong>of</strong>: a) larger body sized<br />
Hierophis viridiflavus in peninsular and continental (i.e. <strong>the</strong> Po valley area, pre-Alpine<br />
areas) Italy, b) smaller body sized Hierophis viridiflavus on small Mediterranean islands<br />
if compared to those with larger body size on large Mediterranean islands and mainland<br />
areas. The main set <strong>of</strong> results clearly indicate a strong differentiation among <strong>the</strong> considered<br />
populations: Hierophis viridiflavus <strong>of</strong> continental and north peninsular Italy do<br />
not differ from <strong>the</strong> populations <strong>of</strong> <strong>the</strong> Ligurian and Tyrrhenian islands. On islands, <strong>the</strong><br />
Whip snakes <strong>of</strong> Corsica and <strong>of</strong> small islands in <strong>the</strong> Tuscan Archipelago display a significantly<br />
high number <strong>of</strong> ventrals while in <strong>the</strong> Sardinian and Sicilian populations a lower<br />
number <strong>of</strong> ventrals has been found; <strong>the</strong> lowest number <strong>of</strong> ventrals is displayed by <strong>the</strong><br />
Sicilian population. These morphological differences suggest <strong>the</strong> occurrence <strong>of</strong> separate
162 BJOta 3/i-a, 2002 SCALI, CORTI, GENTILLI, LUISELLI, RAZZETTI & ZUFFI<br />
evolutionary patterns. Therefore, as a consequence we propose to assign <strong>the</strong> Sardinian<br />
populations to Hierophis viridiflavus sardus (Suckow, 1798) and <strong>the</strong> Sicilian populations<br />
to Hierophis viridiflavus xanthurus (Rafinesque Schmaltz, 1810).<br />
Key words: morphometrics, snakes, Hierophis viridiflavus, Mediterranean distribution<br />
Received 19 October 2001; accepted 6 April 2002<br />
INTRODUCTION<br />
The European Whip snake, Hierophis<br />
viridiflavus, has a southwestern European<br />
distribution, inhabiting a great variety <strong>of</strong><br />
habitats from sea level up to 2000 m a.s.l.<br />
(Heimes 1993, Naulleau 1997). Formerly<br />
divided into three different subspecies,<br />
Hierophis viridiflavus viridiflavus, H.v.carbonarius<br />
and H.v. kratzeri, it has been<br />
quite recently subjected to a taxonomic<br />
revision by Schatti & Vanni (1986): <strong>the</strong>se<br />
authors suggested that most <strong>of</strong> <strong>the</strong> variability<br />
observed is due to very high phenotypic<br />
variation, and <strong>the</strong>refore this<br />
species must be considered monotypic<br />
(Schatti & Vanni 1986, Heimes, 1993).<br />
On <strong>the</strong> o<strong>the</strong>r hand, very recent (in <strong>the</strong><br />
present issue) genetic studies carried out<br />
by Joger and co-workers show that two<br />
clearly different genetic groups could be<br />
identified, a western one occurring in<br />
France, Switzerland and Italy west <strong>of</strong> <strong>the</strong><br />
Apennines, and an eastern one found in<br />
Croatia, Slovenia and eastern Italy (Nagy<br />
et al. 2001). Also, an increasing interest<br />
regarding ecology and biology <strong>of</strong> <strong>the</strong><br />
European whip snake is also now evident<br />
(Ci<strong>of</strong>i & Chelazzi 1991, Capizzi et al.<br />
1995, Capula et al. 1997, Springolo &<br />
Scali 1998, Scali & Montonati 2000, Zuffi<br />
2001 a), but o<strong>the</strong>r aspects <strong>of</strong> life history<br />
traits are still anecdotal (Zuffi 2001 b,<br />
Zuffi et al. 2000). Because <strong>of</strong> <strong>the</strong> wide<br />
distribution <strong>of</strong> this species and its abundance<br />
around <strong>the</strong> Tyrrhenian and<br />
Ligurian Sea coasts, it could be <strong>of</strong> great<br />
interest to test whe<strong>the</strong>r any geographic<br />
variation <strong>of</strong> morphological features could<br />
be related to different ecological adaptations<br />
(i.e.: dietary habits, reproductive<br />
strategies; Zuffi et al. 2000), as in <strong>the</strong><br />
case <strong>of</strong> mainland versus large or small<br />
island populations. Recently, new interest<br />
has been addressed to morphological<br />
variation and ecological plasticity <strong>of</strong> some<br />
Mediterranean reptiles (Delaugerre &<br />
Cheylan 1992, Corti et al 2001, Luiselli &<br />
Zuffi 2002). In particular we focused our<br />
attention on: a) larger body sized H. viridiflavus<br />
<strong>of</strong> continental and peninsular Italy,<br />
b) smaller body sized H. viridiflavus <strong>of</strong><br />
small Mediterranean islands if compared<br />
to those larger body sized specimens <strong>of</strong><br />
large islands and continental areas, in<br />
order to understand which biological<br />
structures could be related to life history<br />
traits, such as reproductive patterns or<br />
dietary habits (Delaugerre & Cheylan<br />
1992, Zuffi 2001 a).<br />
MATERIALS AND METHODS<br />
Whip snakes were examined i) from <strong>the</strong><br />
mainland (Lombardy and Tuscany, continental<br />
and peninsular Italy respectively);<br />
ii) from large islands (Corsica, Sardinia,<br />
Sicily); iii) from small islands (Elba,<br />
Montecristo, Pianosa, Capraia-Tuscan<br />
Archipelago; Tavolara, Caprera, Mai di<br />
Ventre-Sardinian satellite islands) (Figure<br />
1). Standard measurements on 326 specimens<br />
<strong>of</strong> H. viridiflavus from herpetological<br />
collections <strong>of</strong> <strong>the</strong> Universities <strong>of</strong><br />
Florence, Milan and Pisa and from private<br />
collections were considered (± 1 mm):<br />
snout-vent length (SVL), tail length (tL),<br />
head length (HL), head width (HW),<br />
interorbital length (INT-ORB), number <strong>of</strong><br />
ventrals (VS), and number <strong>of</strong> subcaudals<br />
(SCS). Data are presented as average ± 1<br />
SD. The measurements were natural log-
SCALI, CORTI, GENTILLI, LUISELLI, RAZZETTI & ZUFFI Biota 3/1-2, 2002 163<br />
Figure 1. Study area and localities. 1 =<br />
Tuscany; 1bis= Lombardy; 2=Elba;<br />
3=Pianosa; 4=Montecristo; 5=Corsica;<br />
6=Sardinia; 7=Sicily.<br />
transformed to reach linearity and tested<br />
for normality. We used <strong>the</strong> residuals <strong>of</strong><br />
head size parameters against SVL to<br />
avoid any potential effect caused by<br />
snake size; Student t tests were used for<br />
<strong>the</strong> means between sexes. MANOVA was<br />
used for (i) island vs. continent, and (ii)<br />
between all studied localities (only those<br />
with sample size > 4). ANOVA (with<br />
post-hoc multirange tests, LSD test,<br />
Lesser Significant Differences), to highlight<br />
<strong>the</strong> significant group <strong>of</strong> variation<br />
(only those with sample size > 4). Tests<br />
were two-tailed, and set at ( = 0.05, with<br />
SPSS 8.0.<br />
RESULTS<br />
Intersexual comparison<br />
Whip snakes showed strong sexual size<br />
dimorphism in all characters (all with P <<br />
0.01), but HL (P > 0.05), with males evidently<br />
larger and longer (SVL: 819.4 ±<br />
147.5 mm, n = 179; HL: 25 ± 6 mm, n =<br />
116; HW: 16.6 ± 3.6 mm, n = 105; INT-<br />
ORB: 10 + 1.5 mm, n = 91) than females<br />
(SVL: 728.5 ± 147.9 mm, n = 77; HL:<br />
24.4 ± 6.1 mm, n = 54; HW: 13.5 ± 3.5<br />
mm, n = 52; INT-ORB: 8.5 + 1.4 mm, n =<br />
34), and with a lower number <strong>of</strong> VS<br />
(203.8 ± 21.3 vs. 210.5 ± 14.6). TL and<br />
SCS were excluded because <strong>of</strong> <strong>the</strong> high<br />
frequency <strong>of</strong> damaged or injured tails.<br />
Mainland-island comparison<br />
Whip snake females were too scarce<br />
(sample size < 4) from many localities for<br />
any valid statistical approach to be carried<br />
out Whip snake males appeared significantly<br />
different in VS (F = 5.278, P <<br />
0.01, df = 4, R2 = 0.315, Radj = 0.255)<br />
and in one covariate (HL, F = 10.581, P <<br />
0.01, df = 1) between mainland and<br />
island (large plus small islands) categories,<br />
but not because <strong>of</strong> geographic area (F =<br />
3.834, P > 0.05).<br />
All localities comparison (with N > 4)<br />
Whip snake males showed clearcut differences<br />
in VS (F = 4.931, P < 0.001, df = 9;<br />
R2 = 0.52, Radj = 0.414), with no variation<br />
<strong>of</strong> covariates, but a significant influence<br />
<strong>of</strong> considered areas (F = 3.733, P <<br />
0.01). Among <strong>the</strong>se, snakes from<br />
Montecristo, Pianosa, Elba and Corsica<br />
have a significantly higher number <strong>of</strong><br />
ventral scales if compared to those from<br />
Sardinia and Sicily (Figure 2).<br />
Fur<strong>the</strong>rmore, <strong>the</strong> latter show <strong>the</strong> lowest<br />
number <strong>of</strong> ventral scales <strong>of</strong> <strong>the</strong> Whip<br />
snake populations considered.<br />
DISCUSSION<br />
Sexual dimorphism is well documented in<br />
Hierophis viridiflavus, with larger and<br />
longer males than females (Springolo &<br />
Scali 1998, Scali & Montonati 2000). It<br />
has been long questioned whe<strong>the</strong>r <strong>the</strong>re<br />
are possible advantages in having sexes<br />
differ in head or body size. Shine (1994)<br />
argued that <strong>the</strong> larger male is a common<br />
pattern in species with evident male-male<br />
combats; Bonnet et al. (1999) found that<br />
Whip snake males are favoured in movement<br />
and long displacements by having a<br />
higher muscle mass if compared to
Biota 3/1-2, 2002 SCALI, CORTI, GENTILLI, LUISELLI, RAZZETTI & ZUFFI<br />
Figure 2. Geographical variation <strong>of</strong> ventral scales (average ± 1 SD) <strong>of</strong> Hierophis viridiflavus<br />
males.<br />
o<br />
M<br />
220<br />
210<br />
200<br />
190<br />
N- 12 5 25 13 16 5 S 16<br />
Tuscany Montecristo Sardinia Pianosa<br />
Elba Corsica Sicily Lombardy<br />
females. The occurrence <strong>of</strong> geographical<br />
variation in Whip snake males in <strong>the</strong><br />
Mediterranean islands considered shows<br />
that j) <strong>the</strong> high number <strong>of</strong> VS found in<br />
Montecristo, Pianosa and Corsica and <strong>the</strong><br />
low number <strong>of</strong> VS found in Sardinia and<br />
Sicily populations respectively are not<br />
correlated to <strong>the</strong> area size, ii) <strong>the</strong><br />
observed variability <strong>of</strong> Hierophis viridiflavus<br />
follows a north-south oriented pattern.<br />
This variability in Hierophis viridiflavus<br />
shares <strong>the</strong> same zoogeographical<br />
history <strong>of</strong> many Mediterranean animal<br />
populations (Corti et al. 1991, Masseti<br />
1993). Island snakes could have colonised<br />
<strong>the</strong> whole area during <strong>the</strong> Messinian<br />
salinity crisis (6 MY). They may have differentiated<br />
during several climatic<br />
changes according to independent<br />
trends. Sicilian whip snakes resemble <strong>the</strong><br />
snakes <strong>of</strong> sou<strong>the</strong>rn Italy, such as those in<br />
Calabria and Apulia (unpubl. data), with<br />
constant melanism, reduced body size<br />
and significant low number <strong>of</strong> ventral and<br />
caudal scales. The pattern <strong>of</strong> scale variation<br />
underlines an actual status <strong>of</strong> differ-<br />
ent body size structure (Saint Girons<br />
1978, Padoa 1981), a genetically determined<br />
character (Shine 2000). This significant<br />
difference throughout <strong>the</strong> species<br />
distributive area, particularly evident in<br />
<strong>the</strong> island populations considered, in<br />
accordance with preliminary data sets<br />
and suggestions presented by Nagy et al.<br />
(2001), leads us to reconsider that some<br />
<strong>of</strong> <strong>the</strong> Mediterranean populations <strong>of</strong><br />
Hierophis viridiflavus may be separated<br />
taxonomic units. We <strong>the</strong>refore suggest<br />
that <strong>the</strong> Sardinian populations could be<br />
tentatively assigned to Hierophis viridiflavus<br />
sardus (Suckow, 1798) and <strong>the</strong><br />
Sicilian populations to Hierophis viridiflavus<br />
xanthurus (Rafinesque Schmaltz,<br />
1810) (see Table 1 for morphometric<br />
data), even if a fur<strong>the</strong>r interdisciplinary<br />
approach <strong>of</strong> both morphometric and<br />
molecular techniques (see Nagy et al.<br />
2001) should be a necessary step <strong>of</strong><br />
investigation.
SCALI, CORTI, GENTILLI, LUISELLI, RAZZETTI & ZUFFI 3/1-2, 2OO2 165<br />
Table 1. Morphometric data <strong>of</strong> Sardinian and Sicilian proposed subspecies.<br />
Hierophis viridiflavus sardus<br />
Hierophis viridiflavus xanthurus<br />
sex<br />
male<br />
female<br />
male<br />
SVL<br />
n=16<br />
749.1±96.4<br />
558-860<br />
n=3<br />
656.7±55.9<br />
620-721<br />
n=5<br />
545±215.3<br />
372-860<br />
tL<br />
n=ll<br />
278.2±51.3<br />
192-334<br />
n=3<br />
217.7±27.6<br />
189-244<br />
n=5<br />
176.4±56.6<br />
121-271<br />
VS<br />
n=16<br />
200.2±4.2<br />
193-207<br />
n=3<br />
215.3±4.5<br />
211-220<br />
N=5<br />
195.4±2.3<br />
193-199<br />
Acknowledgements<br />
We thank <strong>the</strong> Italian Ministry <strong>of</strong> Agriculture and <strong>the</strong> National Park "Arcipelago<br />
Toscano" for permission to collect data on <strong>the</strong> Natural Reserve "Isola di A/lontecristo",<br />
<strong>the</strong> EU INTERREGG II project Corse-Toscana (Province <strong>of</strong> Livorno and University <strong>of</strong> Pisa)<br />
for granting most <strong>of</strong> <strong>the</strong> research; <strong>the</strong> Parco Naturale di Migliarino, S.Rossore<br />
Massaciuccoli (Pisa) for permission to enter most protected areas; <strong>the</strong> Parco Naturale<br />
del Ticino and <strong>the</strong> Parco Naturale di Groane in Lombardy for permission to collect data;<br />
and <strong>the</strong> Herpetological Department <strong>of</strong> <strong>the</strong> California Academy <strong>of</strong> Sciences (CAS) <strong>of</strong> San<br />
Francisco (USA), <strong>the</strong> Museums <strong>of</strong> Natural History <strong>of</strong> Milan, Morbegno (Sondrio),<br />
Genua, Florence and Pisa. We also wish to thank all colleagues, friends, co-workers and<br />
students who provided useful information on preserved snakes in private collections.<br />
Finally, we wish to thank <strong>the</strong> Museum <strong>of</strong> Natural History and Territory (University <strong>of</strong><br />
Pisa) for grants to MALZ.<br />
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Martens, H., Martinez Rica, J.P., Maurin, H., Oliveira, M.E., S<strong>of</strong>ianidou, T.S.,<br />
Veith, M., and Zuiderwijk, A.(eds.). Atlas <strong>of</strong> Amphibians and Reptiles in Europe.<br />
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(IEGP/SPN), Paris: 342-343.<br />
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e piante della Sicilia, con varie osservazioni sopra i medesimi. Sanfilippo,<br />
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Europe (Reptilia, Viperidae). Rev. suisse Zool. 85: 565-595.<br />
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di colubrini italiani (Coluber viridiflavus ed Elaphe longissima: Reptilia,<br />
Serpentes, Colubridae) su basi biometriche. Atti I Congresso Nazionale della<br />
Societas Herpetologica Italica, Torino 1996. Mus. reg. Sci. nat. Torino: 429-434.<br />
SCHATTI, B., & VANNI, S. 1986: Intraspecific variation in Coluber viridiflavus Lacepede,<br />
1789, and validity <strong>of</strong> its subspecies (Reptilia, Serpentes, Colubridae). Rev. suisse<br />
Zool., 93: 219-232.<br />
SHINE, R, 1994: Sexual size dimorphisem in snake revisited, Copeia 1994: 326-346.<br />
SHINE, R. 2000: Vertebral numbers in male and female snakes: <strong>the</strong> roles <strong>of</strong> natural, sexual<br />
and fecundity selection. J. Evol. Biol. 13: 455-465.<br />
SPRINGOLO, M. AND SCALI, S. 1998: Sexual dimorphism and ontogenetic changes in<br />
Coluber viridiflavus: a head morphometric analysis. In: Miaud, C. & Guyetant, R.<br />
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Herpetologica. Le Bourget du Lac-France: 413-417.<br />
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der Thiere. Weidmannischen Buchandlung, Leipzig<br />
ZUFFI, M.A.L. 2001 a: Diet and morphometrics <strong>of</strong> Coluber (=Hierophis) viridiflavus on <strong>the</strong><br />
island <strong>of</strong> Montecristo (Tyrrhenian Sea, Italy). Herpetological Journal 11:123-125.<br />
ZUFFI, M.A.L. 2001 b: Morphological and functional variability <strong>of</strong> <strong>the</strong> whip snake (Hierophis<br />
viridiflavus) from Corsica: a comparative approach to o<strong>the</strong>r Mediterranean<br />
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Unione Zoologica Italiana, S. Benedetto del Tronto (AP), 24-28 settembre 2000,<br />
Abstracts, 1 pag.
SUROVA Biota 3/i-a, 2002 167<br />
The role <strong>of</strong> frog egg aggregations<br />
as a control <strong>of</strong> abiotic factors<br />
Galina. S. SUROVA<br />
Moscow State University, Biological Faculty, Department Theory <strong>of</strong> Evolution<br />
E-mail: avs@avs.bio.msu.su<br />
Abstract<br />
Differences in oxygen concentration, temperature and pH between clutches <strong>of</strong> frog egg<br />
aggregations in Rana temporaria and ambient water were estimated. It was suggested<br />
that <strong>the</strong>se parameters could have an effect on egg mortality. We found that oxygen<br />
concentration, temperature and pH inside <strong>the</strong> clutch aggregations are different from<br />
those in <strong>the</strong> ambient water. Usually <strong>the</strong> temperature is higher within clutches so <strong>the</strong><br />
accumulation <strong>of</strong> heat by eggs may have adaptive importance. Oxygen concentration is<br />
lower and pH becomes a little higher at <strong>the</strong> end <strong>of</strong> embryonic life; this last phenomenon<br />
should be <strong>the</strong> result <strong>of</strong> natural physiological processes. Never<strong>the</strong>less, investigative<br />
parameters reflect <strong>the</strong> singularity <strong>of</strong> a particular locality in <strong>the</strong> pond.<br />
Key words: Rana temporaria, clutches; aggregations; temperature; concentration <strong>of</strong><br />
oxygen<br />
Received: 28 February 2002; accepted 17 May 2002
168 Biota 3/i-a, 2002 SUROVA<br />
INTRODUCTION<br />
In <strong>the</strong> middle latitudes brown frogs lay<br />
<strong>the</strong>ir eggs in large aggregations. It is a<br />
current opinion that such aggregations<br />
are condensers <strong>of</strong> heat (Seale 1982) and<br />
protect embryos and tadpoles from<br />
morning frost. The aggregation diminishes<br />
<strong>the</strong> effect <strong>of</strong> daily temperature fluctuations<br />
and prevents <strong>the</strong> overcooling <strong>of</strong><br />
larvae (Waldman 1982). The thick layer<br />
<strong>of</strong> slime protects embryos and still inactive<br />
tadpoles from invertebrate predators<br />
(Rough 1976) and <strong>the</strong> pernicious influence<br />
<strong>of</strong> acid marsh waters (Freda 1986)<br />
Thus, aggregations are a perfect protection<br />
against external factors <strong>of</strong> <strong>the</strong> environment.<br />
The larger <strong>the</strong> aggregation, <strong>the</strong><br />
more strongly expressed its protective<br />
effect is (Waldman 1982).<br />
On <strong>the</strong> o<strong>the</strong>r hand, <strong>the</strong> negative effect <strong>of</strong><br />
aggregation is also well known: individuals<br />
from aggregated clutches die more<br />
<strong>of</strong>ten than those from solitary ones,<br />
because <strong>of</strong> high density (Surova &<br />
Severtzov 1985). The factors affecting<br />
mortality in tadpole aggregations when<br />
<strong>the</strong> larvae become free-swimming are<br />
now well known as an "effect <strong>of</strong> group",<br />
i.e. <strong>the</strong>y have an exclusively biogene origin<br />
(Schwartz et al. 1976). O<strong>the</strong>r factors<br />
<strong>of</strong> tadpole and egg mortality during early<br />
ontogeny have been very poorly investigated.<br />
There is only supposition that<br />
death can be caused by hypoxia and possible<br />
poisoning by <strong>the</strong> eggs' own metabolism<br />
products in <strong>the</strong> limited space <strong>of</strong><br />
aggregation (Surova & Severtzov 1985),<br />
as updating <strong>of</strong> <strong>the</strong> environment occurring<br />
here only by diffusion (Burggren 1985)<br />
can be insufficient. There are no concrete<br />
data in this area.<br />
We undertook an attempt to characterize<br />
temperature, oxygen and acid modes in<br />
large aggregations <strong>of</strong> clutches <strong>of</strong> <strong>the</strong><br />
Common Frog Rana temporaria in two<br />
constant ponds at <strong>the</strong> Biological station <strong>of</strong><br />
Moscow State University near<br />
Zvenigorod, Moscow region, Russia.<br />
MATERIAL AND METHODS<br />
Characteristics <strong>of</strong> habitat<br />
Pond 1 is situated in a water-meadow<br />
near <strong>the</strong> Moscow river and is sunny practically<br />
all <strong>the</strong> time. The shallow places<br />
where <strong>the</strong> clutch aggregations were<br />
located were usually very warm. The<br />
study was carried out on two aggregations<br />
(N1 and N2), located near each<br />
o<strong>the</strong>r. Pond 2 is smaller than 1. It is a part<br />
<strong>of</strong> a wood ravine which retains water and<br />
is very shady. The aggregations <strong>of</strong> clutches<br />
here (N3) were only occasionally<br />
warmed by <strong>the</strong> sun during <strong>the</strong> daytime.<br />
Measurements <strong>of</strong> parameters <strong>of</strong><br />
environment<br />
Measurements began at <strong>the</strong> end <strong>of</strong> <strong>the</strong><br />
spawning period, when most frogs have<br />
already laid <strong>the</strong>ir eggs (stage <strong>of</strong> development<br />
up to <strong>the</strong> middle <strong>of</strong> <strong>the</strong> gastrula).<br />
The data were taken over a period <strong>of</strong> two<br />
weeks. Measurements in both ponds<br />
were stopped when <strong>the</strong> tadpoles reached<br />
<strong>the</strong> stage <strong>of</strong> free-swimming larva (stages<br />
<strong>of</strong> development according to Dabagjan &<br />
Sleptsova (1975)).<br />
The measuring instruments were positioned<br />
at different points <strong>of</strong> an aggregation:<br />
at <strong>the</strong> centre and <strong>the</strong> edges, on <strong>the</strong><br />
surface and at various depths (from 3 up<br />
to,. 9 points .for one measurement).<br />
Simultaneous parameters were measured<br />
in ambient water at a distance 15-20 cm<br />
away from an aggregation on <strong>the</strong> surface<br />
and at <strong>the</strong> same depth (also 3-9 points).<br />
All measurements were taken at approximately<br />
<strong>the</strong> same points every time. The<br />
final results are given on <strong>the</strong> diagrams as<br />
average values on all points for one<br />
measurement. Differences were tested<br />
with a t-test. The acidity was measured<br />
by a portable field pH-meter, temperature<br />
and content <strong>of</strong> oxygen (in mg/l) - by<br />
a portable oxymeter.<br />
RESULTS<br />
In pond 1 spawning began on April 10.
SUROVA Biota 3/1-2,2002 169<br />
Table 1. Characteristics <strong>of</strong> development and pH in <strong>the</strong> aggregations.<br />
Stages <strong>of</strong> development according to Dabagjan & Sleptsova (1975):<br />
0 - beginning <strong>of</strong> cleavage; 7 - middle <strong>of</strong> cleavage; 20 - neurula; 26 - 28 - tail bud; 29<br />
- 30 - hatching; 33 - 37 - external gills; 38 - 39 - beginning <strong>of</strong> development buds <strong>of</strong> <strong>the</strong><br />
legs; 39 - free swimming larvae<br />
N<strong>of</strong><br />
Aggregation<br />
1<br />
2<br />
3<br />
Measurement<br />
date<br />
IV<br />
3V<br />
5V<br />
10V<br />
14V<br />
IV<br />
3V<br />
5V<br />
10V<br />
14V<br />
IV<br />
3V<br />
5V<br />
10 V<br />
14V<br />
Age (days)<br />
17<br />
19<br />
22<br />
28<br />
32<br />
17<br />
19<br />
22<br />
28<br />
32<br />
5<br />
7<br />
9<br />
14<br />
18<br />
By <strong>the</strong> 17th <strong>of</strong> April several aggregations<br />
<strong>of</strong> clutches had formed. Two <strong>of</strong> <strong>the</strong>m, situated<br />
in <strong>the</strong> warmest part <strong>of</strong> <strong>the</strong> pond,<br />
were taken for study. Aggregation 1 had<br />
a total area <strong>of</strong> 1.25 m2, number <strong>of</strong> clutches<br />
- 484, density - 387.2 clutches/m2.<br />
Aggregation 2 differed little in size (1.05<br />
m2), but it had a smaller number <strong>of</strong><br />
clutches (174), as a near-bottom layer<br />
was absent, and consequently <strong>the</strong>ir density<br />
was much smaller: (165.7 cl./m2). In<br />
<strong>the</strong> colder pond #2 spawning began on<br />
April 15, and a single, but very large<br />
aggregation was formed only by May 1,<br />
with an area <strong>of</strong> about 6 m2. (1.5m _ 4m),<br />
number <strong>of</strong> clutches - 611, and density -<br />
101.8 cl./m2.<br />
The water in pond 1 was neutral all <strong>the</strong><br />
time; in pond 2 it was alkaline (Table 1).<br />
The pH value in aggregations follows <strong>the</strong><br />
same pattern, i.e. <strong>the</strong> acid mode in aggregations<br />
<strong>of</strong> clutches corresponds to <strong>the</strong><br />
PH<br />
in <strong>the</strong> water<br />
6.9<br />
7.0<br />
7.0<br />
7.0<br />
-<br />
-<br />
7.1<br />
7.0<br />
6.9<br />
-<br />
-<br />
7.8<br />
7.8<br />
7.7<br />
-<br />
pH in <strong>the</strong><br />
aggregation<br />
8.2<br />
6.9<br />
6.8<br />
-<br />
-<br />
-<br />
7.7<br />
6.9<br />
6.7<br />
-<br />
-<br />
7.9<br />
7.7<br />
7.6<br />
-<br />
Stages <strong>of</strong><br />
development<br />
27-28<br />
29-30<br />
33-37<br />
38-39<br />
39<br />
27<br />
29-30<br />
33-37<br />
38-39<br />
39<br />
0-13<br />
7-20<br />
26-28<br />
29-30<br />
33-37<br />
acid mode <strong>of</strong> <strong>the</strong> pond. However, after<br />
hatching, <strong>the</strong> medium in aggregations<br />
became a little bit more acid. This is obviously<br />
connected with <strong>the</strong> beginning <strong>of</strong><br />
active feeding and excretion <strong>of</strong> products<br />
<strong>of</strong> metabolism. pH values in <strong>the</strong> pond and<br />
in aggregations are favourable and are far<br />
from <strong>the</strong> threshold value <strong>of</strong> acidity conducive<br />
to deviations <strong>of</strong> development and<br />
embryo mortality (pH <strong>of</strong> 5.0 and lower;<br />
Freda 1986). The temperature and oxygen<br />
mode changes are given on <strong>the</strong> diagrams<br />
(Figure 1 - 3). In both ponds, temperature<br />
in aggregations was 1-3<br />
degrees higher than in ambient water.<br />
This is especially appreciable in aggregation<br />
2, where <strong>the</strong> difference is significant<br />
through <strong>the</strong> whole ontogeny period<br />
(Figure 2). In aggregation 1 <strong>the</strong> significant<br />
differences were observed only at<br />
<strong>the</strong> stage <strong>of</strong> hatching and at <strong>the</strong> stage <strong>of</strong><br />
free-swimming larva in a very dense,
170 'Biota 3/1-2,2002 SUROVA<br />
Figure 1. Temperature and oxygen content<br />
in ambient water and in <strong>the</strong> aggregation<br />
N 1.<br />
Legend: temperature; 1- in aggregation,<br />
2 - in <strong>the</strong> water;<br />
Oxygen; 3 - in aggregation, 4 - in <strong>the</strong><br />
water, * - p < 0.05; ** - p < 0.01.<br />
May. I May. 3 May. 5 May. 7 May.9 May II May. 13<br />
Figure 2. Temperature and oxygen content<br />
in ambient water and in <strong>the</strong> aggregation<br />
N 2.<br />
For legend see figure 1.<br />
May, I May. 3 May, 5 May. 7 May. 9 May 11 May. 13<br />
Figure 3. Temperature and oxygen content<br />
in ambient water and in <strong>the</strong> aggregation<br />
N 3.<br />
For legend see figure 1.<br />
May. 1 May. 3 May, 5 May, 7 May, 9 May II May. 13<br />
obviously <strong>the</strong>rmal aggregation <strong>of</strong> tadpoles<br />
(Figure 1). In <strong>the</strong> cooler pond 2, <strong>the</strong><br />
temperature in <strong>the</strong> aggregation was significantly<br />
higher than in ambient water<br />
(Figure 3). This relation was reversed on<br />
May 10. It happened because <strong>the</strong> temperature<br />
fell dramatically in a short time.<br />
The temperature in <strong>the</strong> ponds became<br />
lower, but in pond 2 it cooled down much<br />
more strongly (up to 8.7°C, but in pond 1<br />
upto10.5°C).<br />
The change <strong>of</strong> oxygen mode in aggregations<br />
also has a general tendency: <strong>the</strong><br />
level <strong>of</strong> oxygen in aggregations <strong>of</strong> clutches<br />
is always lower than in ambient water.<br />
In aggregation 2 <strong>the</strong>se differences are<br />
significant for all measurements (Figure<br />
2). In aggregation 1, at <strong>the</strong> egg stage and<br />
at hatching <strong>the</strong> level <strong>of</strong> oxygen in clutches<br />
and in <strong>the</strong> surrounding water has no<br />
significant differences, although <strong>the</strong><br />
means are very different (see Figure 1).<br />
After hatching <strong>the</strong> deviation becomes<br />
less, and <strong>the</strong> difference in oxygen content<br />
in aggregations and ambient water<br />
becomes significant. In aggregation 3 <strong>the</strong><br />
oxygen level was different at all times<br />
except at <strong>the</strong> stage <strong>of</strong> tail budding (May<br />
5), when <strong>the</strong> level <strong>of</strong> oxygen in <strong>the</strong><br />
aggregation and in <strong>the</strong> water was practically<br />
equal (Figure 3). On <strong>the</strong> same diagram<br />
it is clearly visible that at a practically<br />
constant level <strong>of</strong> oxygen saturation<br />
in <strong>the</strong> water from May 5 up to May 14,<br />
<strong>the</strong> oxygen level in an aggregation <strong>of</strong><br />
clutches, and <strong>the</strong>n tadpoles, is steadily<br />
reduced, reflecting increasing consumption<br />
<strong>of</strong> oxygen during <strong>the</strong> process <strong>of</strong><br />
development.<br />
DISCUSSION<br />
The consumption <strong>of</strong> oxygen by embryo<br />
and larvae considerably reduces its level<br />
in <strong>the</strong> aggregation. On one hand, our<br />
study has obviously shown that <strong>the</strong> living<br />
conditions <strong>of</strong> aggregation are closely connected<br />
with ambient values. On <strong>the</strong> o<strong>the</strong>r<br />
hand, <strong>the</strong> aggregations <strong>of</strong> clutches form-
SUROVA 3/1-2, 2OO2 171<br />
ing thick layers <strong>of</strong> mucilage, and after<br />
hatching - aggregations <strong>of</strong> larvae with<br />
high density, are an independent system<br />
with temperature higher than ambient, a<br />
little more acidic medium and a lower<br />
content <strong>of</strong> oxygen. The accumulation <strong>of</strong><br />
heat by eggs and dark coloured tadpoles<br />
has obvious adaptive importance, as was<br />
already mentioned above.<br />
Such a strong downturn <strong>of</strong> temperature<br />
as occurred in pond 2 on May 10 could<br />
probably not be compensated for by any<br />
accumulation <strong>of</strong> heat in an aggregation.<br />
This indicates that <strong>the</strong> heat-sink ability <strong>of</strong><br />
an egg mass is not absolute; it has some<br />
limits which depend on ambient temperature.<br />
In o<strong>the</strong>r words, <strong>the</strong> higher <strong>the</strong> temperature,<br />
<strong>the</strong> better expressed is <strong>the</strong> ability<br />
<strong>of</strong> an aggregation to accumulate and<br />
to keep heat.<br />
The o<strong>the</strong>r parameters we tested probably<br />
have no direct adaptive importance but<br />
most likely reflect natural physiological<br />
processes in embryos and larvae. Such a<br />
small increase <strong>of</strong> acidity as we observed<br />
can not render oppressive action upon<br />
growth and development. If this does<br />
occur, it is more likely because <strong>of</strong> excretion<br />
specific metabolites under a high<br />
density <strong>of</strong> tadpoles (Schwartz et al.<br />
1976), but <strong>the</strong> pH level is not affected by<br />
this factor.<br />
The essential decrease <strong>of</strong> ambient oxygen<br />
level - especially just after hatching - can<br />
result in hypoxia and delay <strong>of</strong> ontogeny.<br />
However, it is necessary to note that<br />
hypoxia, naturally arising in aggregations,<br />
serves as a physiological trigger for hatching<br />
(Petranka et al. 1982). Certainly, oxygen<br />
level and pH which conduce to<br />
oppression can only be shown in fur<strong>the</strong>r<br />
laboratory and field experiments. But <strong>the</strong><br />
data given here show that <strong>the</strong> fluctuations<br />
<strong>of</strong> temperature, oxygen and pH in<br />
aggregations do not go beyond <strong>the</strong> limits<br />
capable <strong>of</strong> causing <strong>the</strong> death <strong>of</strong> larvae.<br />
Never<strong>the</strong>less, our earlier study showed<br />
that <strong>the</strong> presence <strong>of</strong> aggregations could<br />
result in 70% larvae mortality (Surova &<br />
Severtzov 1985). Thus, <strong>the</strong> role <strong>of</strong> aggregation<br />
in <strong>the</strong> life <strong>of</strong> embryos and larvae is<br />
ambiguous and encourages us to continue<br />
to study <strong>the</strong> balance between <strong>the</strong><br />
adaptive protective effect <strong>of</strong> aggregations<br />
and <strong>the</strong> high price for selection <strong>of</strong> resistance<br />
to high density.<br />
Conclusions<br />
1. In aggregations <strong>the</strong> temperature is<br />
always higher than in ambient water.<br />
However, during a long and strong<br />
enough decrease <strong>of</strong> temperature, <strong>the</strong><br />
aggregation can not effectively retain<br />
heat.<br />
2. The level <strong>of</strong> oxygen in an aggregation<br />
is always lower than in ambient water.<br />
The content <strong>of</strong> oxygen in an aggregation<br />
is constantly reduced with <strong>the</strong> passage <strong>of</strong><br />
time. This is not always connected to parallel<br />
changes in environment, but more<br />
likely by its consumption by larvae.<br />
3. The acidity in an aggregation does not<br />
differ greatly from <strong>the</strong> acidity in water,<br />
only increasing slightly during late stages<br />
<strong>of</strong> development.<br />
4. The changes <strong>of</strong> investigated parameters<br />
in an aggregation are closely connected<br />
to <strong>the</strong>ir changes in ambient water.<br />
They have <strong>the</strong>ir own character for <strong>the</strong><br />
particular pond and place <strong>of</strong> aggregation<br />
locality.<br />
5. Never<strong>the</strong>less, an aggregation is a more<br />
or less independent system. It has <strong>the</strong><br />
original pr<strong>of</strong>ile <strong>of</strong> parameters <strong>of</strong> internal<br />
abiotic factors that, in a number <strong>of</strong> cases,<br />
can have adaptive importance.
172 Biota 3/i-2, SUROVA<br />
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PETRANKA, J.W., INST, J.I. & CRAWFORD, E.G. 1982: Hatching <strong>of</strong> amphibian embryos: <strong>the</strong><br />
physiological trigger. Science 217: 257-259.<br />
POUCH, F.H. 1976: Acid precipitation and embryonic mortality <strong>of</strong> spotted salamanders,<br />
Ambystoma maculatum. Science 192: 68-70.<br />
SEALE, D.B. 1982: Physical factors influencing oviposition by <strong>the</strong> woodfrog, Rana sylvatica,<br />
in Pennsylvania. Copeia 3: 627-635.<br />
WALDMAN, B. 1982: Adaptive significance <strong>of</strong> communal oviposition in woodfrogs (Rana<br />
sylvatica). Behav. Ecol. and Sociobiol. 10: 169-174.
YOROS, KORSOS & SZALAY _ BlOta 3/1-2, 2003 _ 173<br />
A comparative morphological<br />
study <strong>of</strong> <strong>the</strong> two Hungarian<br />
discoglossid toad species<br />
Bombina spp<br />
Judit VOROS1, Zoltan KORSOS1 &<br />
Ferenc SZALAY2<br />
'Hungarian Natural History Museum, Baross u. 13, H-1088 Budapest, Hungary<br />
E-mail: jvoros@zoo.zoo.nhmus.hu, korsos@zoo.zoo.nhmus.hu<br />
2Szent Istvan University, Faculty <strong>of</strong> Veterinary Science, Department <strong>of</strong> Anatomy and<br />
Histology, Istvan u. 2, H-1077 Budapest, Hungary<br />
E-mail: fszalay@univet.hu<br />
Abstract<br />
In order to study <strong>the</strong> hybridization <strong>of</strong> <strong>the</strong> two Hungarian discoglossid toad species (Firebellied<br />
Toad, Bombina bombina and Yellow-bellied Toad, Bombina variegata), external<br />
morphological features (body length, head length, head width, forelimb length, femur<br />
length, tibia length, and foot length) <strong>of</strong> 271 specimens from 18 different localities <strong>of</strong><br />
Hungary were examined. In <strong>the</strong> hybrid zone (located in <strong>the</strong> nor<strong>the</strong>rn region <strong>of</strong><br />
Hungary) between <strong>the</strong> characteristic ranges <strong>of</strong> <strong>the</strong> two species, intermediate individuals<br />
can be found showing intermediate morphological characters. A new videomorphological<br />
method was developed to quantify <strong>the</strong> external morphological characters, mainly<br />
<strong>the</strong> colour pattern on <strong>the</strong> belly (circularity, mean area, and mean circumference <strong>of</strong><br />
patches, patch density, area ratio, ratio <strong>of</strong> mean area and mean circumference, colour<br />
density), in order to facilitate reliable identification <strong>of</strong> pure and hybrid specimens. With<br />
<strong>the</strong> examination <strong>of</strong> specimens collected from different regions <strong>of</strong> <strong>the</strong> country, a hierarchical<br />
phenetic classification was performed to demonstrate <strong>the</strong> situation <strong>of</strong> <strong>the</strong><br />
Bombina hybrid zones in Hungary.<br />
Key words: Bombina bombina, Bombina variegata, hybridization, morphological characters,<br />
videoanalysis, colour pattern, phenetic classification<br />
Received 30 November 2001'; accepted 12 February 2002
174 3/1-2, 2OO2 VOROS, KORSOS & SZALAY<br />
INTRODUCTION<br />
Natural hybridization between <strong>the</strong> two<br />
European toad species, Fire-bellied Toad<br />
Bombina bombina and Yellow-bellied<br />
Toad Bombina variegata, was first reported<br />
in Hungary by Mehely (1892). The<br />
ranges <strong>of</strong> Fire-bellied Toad and Yellowbellied<br />
Toad meet in a wide front extending<br />
from Austria along <strong>the</strong> sou<strong>the</strong>rn edge<br />
<strong>of</strong> <strong>the</strong> Danube Valley to <strong>the</strong> Black Sea and<br />
surrounding <strong>the</strong> Carpathian Mountains<br />
along <strong>the</strong>ir foothills (Szymura 1993).<br />
Hungary is an interesting part <strong>of</strong> <strong>the</strong><br />
Bombina hybrid zones. In <strong>the</strong> Great Plain,<br />
Fire-bellied Toad is very common, occuring<br />
in most <strong>the</strong> ponds. However, only isolated<br />
Yellow-bellied Toad populations can<br />
be found in <strong>the</strong> hilly regions <strong>of</strong> <strong>the</strong> country<br />
(above 300 m a.s.l.). In <strong>the</strong> regions<br />
where <strong>the</strong> ranges <strong>of</strong> <strong>the</strong> two species<br />
meet, <strong>the</strong>y interbreed and form hybrid<br />
populations. Bombina hybridization in<br />
Hungary has been investigated in detail in<br />
<strong>the</strong> Matra Mountains and <strong>the</strong> Aggtelek<br />
Karst by Gollmann (1987 and 1986,<br />
respectively). It has been reported from<br />
<strong>the</strong> Mecsek Mountains (Mehely 1904),<br />
<strong>the</strong> Biikk Mountains (Dely 1996), and<br />
observed in <strong>the</strong> Zemplen Mountains as<br />
well (Voros 2000).<br />
In our study we investigated individual<br />
species identification <strong>of</strong> Bombina populations<br />
throughout Hungary, using external<br />
morphological characters. We compared<br />
samples from populations <strong>of</strong> both <strong>the</strong><br />
typical habitats and from those with<br />
intermediate characters.<br />
We examined whe<strong>the</strong>r hybrid populations<br />
can be found in <strong>the</strong> presumed<br />
hybrid zones and what percentage <strong>of</strong> <strong>the</strong><br />
measured specimens can be considered<br />
as hybrid. We also determined <strong>the</strong> most<br />
reliable character among 12 for identification<br />
<strong>of</strong> <strong>the</strong> two Bombina species.<br />
MATERIAL AND METHODS<br />
Our study area contained 18 different<br />
study localities in Hungary (Figure 1).<br />
These were both typical Fire-bellied Toad<br />
and Yellow-bellied Toad habitats.<br />
Localities where <strong>the</strong> ranges <strong>of</strong> <strong>the</strong> two<br />
species meet, and hence interbreed, were<br />
preferred when selecting sampling sites.<br />
Altoge<strong>the</strong>r 271 Bombina specimens were<br />
studied, using two morphological methods.<br />
Seven morphometric characters<br />
were measured in <strong>the</strong> field, with an accuracy<br />
<strong>of</strong> 0.1 millimetre: body length, head<br />
length, head width, forelimb length,<br />
femur length, tibia length, and foot<br />
length.<br />
A short video recording was shot in <strong>the</strong><br />
field <strong>of</strong> <strong>the</strong> ventral surface <strong>of</strong> every specimen,<br />
which was later analysed by a<br />
newly developed computer program to<br />
describe details <strong>of</strong> <strong>the</strong> colour pattern <strong>of</strong><br />
<strong>the</strong> belly. This method was chosen<br />
because, superficially, <strong>the</strong> two species can<br />
be distinguished most easily by observations<br />
<strong>of</strong> <strong>the</strong> colour and pattern <strong>of</strong> <strong>the</strong><br />
ventral surface. Video tapes were played<br />
and paused at each individual. A rectangle<br />
on <strong>the</strong> belly <strong>of</strong> <strong>the</strong> specimens<br />
(between <strong>the</strong> four limbs) was selected by<br />
<strong>the</strong> program, and this part <strong>of</strong> <strong>the</strong> ventral<br />
surface was analysed. The exact characters<br />
calculated by <strong>the</strong> computer program<br />
were: circularity, mean area, and mean<br />
circumference <strong>of</strong> patches, patch density,<br />
area ratio (ratio <strong>of</strong> <strong>the</strong> colour<br />
patches/background), ratio <strong>of</strong> mean area<br />
and mean circumference, and colour density.<br />
The computer program will be<br />
described in detail and published separately,<br />
and will be available in <strong>the</strong> future.<br />
Multivariate discriminant analyses were<br />
carried out for morphometric and videomorphological<br />
characters, first separated,<br />
<strong>the</strong>n combined toge<strong>the</strong>r. Before <strong>the</strong><br />
analysis we grouped all specimens into<br />
Fire-bellied Toad or Yellow-bellied Toad<br />
species according to <strong>the</strong> classical morphological<br />
methods (Mehely 1891). There<br />
were geographical regions <strong>of</strong> Hungary<br />
where we could find clearly identifiable<br />
individuals, such as <strong>the</strong> Hortobagy,
VORC3S, KORSOS & SZALAY Biota 3/i-a, 2002 175<br />
Tihany, and Dinnyes for Fire-bellied Toad,<br />
and <strong>the</strong> Mecsek Mts. for Yellow-bellied<br />
Toad, and we could compare <strong>the</strong> populations<br />
from <strong>the</strong>se regions to <strong>the</strong> o<strong>the</strong>rs. A<br />
phenetic classification (unweighted pairgroup<br />
average method with Euclidean<br />
distances) <strong>of</strong> <strong>the</strong> 18 sampling sites was<br />
applied to define <strong>the</strong> hierarchical relation<br />
<strong>of</strong> <strong>the</strong> populations. All analyses were carried<br />
out using Statistica for Windows<br />
(Version 5).<br />
RESULTS<br />
Discriminant analysis<br />
A discriminant analysis was applied for<br />
<strong>the</strong> two feature group (morphometric<br />
and videomorphological), first separated<br />
and <strong>the</strong>n toge<strong>the</strong>r. The first part <strong>of</strong> <strong>the</strong><br />
results shows <strong>the</strong> significance <strong>of</strong> <strong>the</strong> characters<br />
in question affecting <strong>the</strong> segregation<br />
<strong>of</strong> <strong>the</strong> groups.<br />
ed in <strong>the</strong> mean circumference <strong>of</strong> <strong>the</strong><br />
patches, area ratio, colour density, forelimb<br />
length, and tibia length (Table 1).<br />
The o<strong>the</strong>r result <strong>of</strong> <strong>the</strong> discriminant analysis<br />
was <strong>the</strong> misclassification probability, in<br />
case <strong>of</strong> identification based on classical<br />
morphological features, into two species.<br />
The classification results show that preidentification<br />
with <strong>the</strong> classical morphological<br />
method corresponds by 92.46%<br />
to identification with <strong>the</strong> computer program<br />
based on <strong>the</strong> analysis <strong>of</strong> <strong>the</strong> colour<br />
patterns <strong>of</strong> <strong>the</strong> belly (Figure 2), but by<br />
only 87.44% with morphometric measurements<br />
(Figure 3.). If we take into consideration<br />
all 12 features, <strong>the</strong> correspondence<br />
is 95.48% (Figure 4).<br />
At <strong>the</strong> same time, <strong>the</strong> misclassification<br />
probabilities show <strong>the</strong> overlap between<br />
<strong>the</strong> two pre-identified groups (i.e. <strong>the</strong><br />
two species). These percentages (7.54%,<br />
Table 1. The significance and <strong>the</strong> Wilks' Lambda values <strong>of</strong> <strong>the</strong> 12 characters.<br />
Characters<br />
Mean area<br />
Mean circumference<br />
Patch density<br />
Area ratio<br />
Ratio <strong>of</strong> <strong>the</strong> area and circ.<br />
Colour density<br />
Circularity<br />
Head length/ head width<br />
Forelimb length<br />
Femur length<br />
Tibia length<br />
Foot length<br />
According to <strong>the</strong> significance and <strong>the</strong><br />
Wilks' Lambda values (Wilks' Lambda is a<br />
statistic for <strong>the</strong> differentiation <strong>of</strong> two statistical<br />
populations; its value is 1 if <strong>the</strong> centroids<br />
<strong>of</strong> <strong>the</strong> two groups are indistinguishable,<br />
and 0 at maximal divergence (Podani<br />
1997), significant differences were detect-<br />
p-values<br />
0.326<br />
0.0001<br />
0.136<br />
0.0003<br />
0.224<br />
0.0003<br />
0.143<br />
0.259<br />
0.001<br />
0.027<br />
0.01<br />
0.117<br />
Wilks' lambda values<br />
0.246<br />
0.269<br />
0.248<br />
0.262<br />
0.247<br />
0.262<br />
0.248<br />
0.247<br />
0.259<br />
0.252<br />
0.254<br />
0.248<br />
12.56%, 4.52%) indicate <strong>the</strong> misclassification<br />
<strong>of</strong> <strong>the</strong> videomorphological method<br />
(Figure 2-4).<br />
Phenetic classification<br />
To understand <strong>the</strong> relationships between<br />
<strong>the</strong> populations, we made a phenetic
176 Biota 3/1-2,2002 VOROS, KORS6S & SZALAY<br />
Figure 1. Sampling localities: 1. Aggtelek,<br />
2. Zemplen, 3. Beregi Plains, 4.<br />
Hortobagy, 5. Matra, 6. Kiskunsag, 7.<br />
Pilis, 8. Vertes, 9. Lake Velence, 10. River<br />
Drava, 11. Mecsek, 12. Lake Kis-Balaton,<br />
13. Balatonfenyves, 14. Tihanyi<br />
Peninsula, 15. Bakony, 16. Orseg, 17.<br />
Hansag, 18. Szigetkoz.<br />
classification <strong>of</strong> <strong>the</strong> samples according to<br />
locality (samples from 38 populations).<br />
The phenogram (Figure 5) gave a hierarchical<br />
connection clearly defining <strong>the</strong> two<br />
Bombina species. There are specimens<br />
from closely situated study plots which<br />
belong to different species.<br />
DISCUSSION<br />
Among <strong>the</strong> videomorphological characters,<br />
<strong>the</strong> mean circumference <strong>of</strong> patches,<br />
<strong>the</strong> area ratio, and colour density<br />
(p=0.0001, 0.0003, 0.0003), and among<br />
<strong>the</strong> morphometric characters, <strong>the</strong> forelimb<br />
length and tibia length (p= 0.0027,<br />
0.01) were found to be most reliable for<br />
<strong>the</strong> differentiation <strong>of</strong> <strong>the</strong> two species.<br />
Regarding a single individual, consideration<br />
<strong>of</strong> all 12 characters proved to be <strong>the</strong><br />
most reliable method for species identification.<br />
According to our study <strong>of</strong> <strong>the</strong> Aggtelek<br />
region (north-eastern Hungary) <strong>the</strong>re are<br />
hybrid populations. The samples <strong>of</strong><br />
Aggtelek were taken from closely situated<br />
sites, and because <strong>the</strong> specimens<br />
belong to both groups (species), <strong>the</strong>y<br />
have intermediate morphological characters.<br />
The ranges <strong>of</strong> <strong>the</strong> two species meet<br />
here and individuals interbreed in <strong>the</strong>se<br />
habitats. The Pilis Hill (close to Budapest)<br />
also appeared to be an interesting locality,<br />
since specimens from this site had an<br />
uncertain position in <strong>the</strong> phenogram.<br />
To be able to define <strong>the</strong> hybrid zones <strong>of</strong><br />
Bombina species, we would like to study<br />
<strong>the</strong> genetic background <strong>of</strong> <strong>the</strong> morphological<br />
characters in <strong>the</strong> future.<br />
Figure 2. The overlap in case <strong>of</strong> <strong>the</strong> analysis <strong>of</strong> colour patches <strong>of</strong> <strong>the</strong> belly. The striped<br />
columns show Fire-bellied Toad Bombina bombina, <strong>the</strong> blank columns show Bombina<br />
variegata. Overlap (misclassification probability) 7.54%.<br />
30<br />
o 15<br />
N N<br />
-4,5 -4,0 -3,5 -3,0 -2,5 -2,0 -1,5 -1,0 -0,5 0,0 0,5 1,0 1,5 2,0 2,5 3.0 3.5 4,0
VOROS, KORSOS & SZALAY Biota 3/i-a, 2002 177<br />
Figure 3. The overlap in case <strong>of</strong> <strong>the</strong> morphometric measurements. The striped columns<br />
show Fire-bellied Toad Bombina bombina, <strong>the</strong> blank columns show Bombina variegata.<br />
Overlap (misclassification probability) 12.56%.<br />
-5,0 -5,0 -4,0 -3,0 -2,0 -1,0 0,0 1,0 2,0 3,0 4,0<br />
-5,5 -4,5 -3,5 -2,5 -1,5 -0,5 0,5 1,5 2,5 3,5 4,5<br />
Figure 4. The overlap in case <strong>of</strong> analysis by all 12 features. The blank columns show<br />
Fire-bellied Toad Bombina bombina, <strong>the</strong> striped columns show Bombina variegata.<br />
Overlap (misclassification probability) 4.55%.<br />
-5.5 -4.5 -3,5 -2,5 -1,5 -0.5 0,5 1,5 2,5 3,5<br />
-5,0 -4,0 -3,0 -2,0 -1,0 0,0 1,0 2,0 3,0<br />
Expected<br />
Normal<br />
Expected<br />
Normal
178 Biota 3/1-2,2002 VOROS, KORSOS & SZALAY<br />
Figure 5. Phenogram (UPGMA, Euclidean distances) <strong>of</strong> <strong>the</strong> samples according to localities.<br />
The upper group corresponds to Fire-bellied Toad Bombina bombina, <strong>the</strong> lower<br />
group to Bombina variegata.<br />
Uertes<br />
Plaiul<br />
Hortobag<br />
T ihany<br />
Dinnyes<br />
ftggtell<br />
T iszahat<br />
Szerenle<br />
Hortob2<br />
Baja<br />
Kisbal<br />
Mariaf<br />
Hortobl<br />
Ki skunsa<br />
Taljand<br />
Sz iget 1<br />
Bakony<br />
Hansag<br />
Sz iget 2<br />
Pilis<br />
Ronania<br />
Matra<br />
Ujhutal<br />
UjhutaS<br />
Ujhuta4<br />
Regec<br />
flggtelS<br />
Ohuta<br />
Ujhuta2<br />
Gone 2<br />
Goncl<br />
Zenplenl-<br />
Mecsek i<br />
Mecsek 2<br />
Ujhuta3<br />
UjhutaG<br />
Acknowledgements<br />
We would like to thank <strong>the</strong> Nature Conservation Directorates <strong>of</strong> <strong>the</strong> Balaton Upland<br />
and <strong>the</strong> Duna-lpoly National Parks for granting study permits in <strong>the</strong>ir respective territories.<br />
We are grateful to numerous persons not listed here who have made possible <strong>the</strong><br />
collection <strong>of</strong> field data, and, fur<strong>the</strong>rmore, especially to Giinter Gollmann (University <strong>of</strong><br />
Vienna) for sharing his literature, and to Gabor Herczeg and Mihaly Foldvari (Budapest)<br />
for technical support.<br />
REFERENCES<br />
DELY, O. GY. 1996: Amphibians and reptiles <strong>of</strong> <strong>the</strong> Biikk Mountains. In: Mahunka, S. (ed):<br />
The fauna <strong>of</strong> <strong>the</strong> Bukk National Park. Vol. 2. Hungarian Natural History Museum,<br />
Budapest: 535-572.<br />
GOLLMANN, G. 1987: Bombina bombina and Bombina variegata in <strong>the</strong> Matra mountains:<br />
New data on distribution and hybridization. Amphibia-Reptilia 8: 213-224.<br />
f GOLLMANN, G., ROTH, P. & HODL, W. 1986: Hybridization between <strong>the</strong> Fire-bellied toads<br />
Bombina bombina and Bombina variegata in <strong>the</strong> karst regions <strong>of</strong> Slovakia and<br />
Hungary: morphological and allozyme evidence. Journal <strong>of</strong> Evolutionary Biology<br />
1:3-14.<br />
MEHELY, L. 1891: A magyar fauna Bombinatorjai es egy uj Triton (Molge) faj hazankbol.<br />
MTA Ma<strong>the</strong>matikai es Termeszettudomanyi Kozlemenyek 24: 553-574. (in
VOROS, KORSOS & SZALAY BJOta 3/1-2,2002 179<br />
Hungarian)<br />
MERELY, L. 1892: Beitrage zur Kenntnis der Bombinator-Arten, sowie deren Standorte und<br />
Verbreitung in Ungarn. Ma<strong>the</strong>matische und Naturwissenschaftliche Berichte aus<br />
Ungarn 10: 55-79.<br />
MF.HELY, L. 1904: A Mecsekhegyseg es a Kapela herpetologiai viszonyai. Allattarvi<br />
Kozlemengek 3: 241-289. In Hungarian.<br />
PODANI, J. 1997: Bevezetes a tobbvaltozos biologiai adatfeltaras rejtelmeibe. -Budapest,<br />
412 pp. In Hungarian<br />
STATISTICA FOR WINDOWS, Version 5., StatS<strong>of</strong>t, Inc.<br />
SZYMURA, J. M. 1993: Analysis <strong>of</strong> hybrid zones with Bombina. In: Harrison, R. G. (ed.).<br />
Hybrid zones and <strong>the</strong> evolutionary process. Oxford University Press, New York,<br />
Oxford, pp. 261-289.<br />
VOROS, J. 2000: A comparative morphological study on Hungarian Fire-bellied and Yellowbellied<br />
toad (Bombina spp.) populations. M. Sc. Thesis, Szent Istvan University,<br />
Faculty <strong>of</strong> Veterinary Science, Institut for Zoology, Budapest, 38 pp. In Hungarian.
ZAVADIL & SIZLING Biota 3/1-2. 2002 181<br />
Morphological variability in <strong>the</strong><br />
newts <strong>of</strong> <strong>the</strong> Cristatus group<br />
Vit ZAVADIL1 & Arnost L SIZLING1,2<br />
Agency for Nature Conservation and Landscape Protection <strong>of</strong> <strong>the</strong> Czech Republic,<br />
Kalicnicka 4-6, Praha 3, 130 00, Czech Republic<br />
E-mail: zavadil@nature.cz,<br />
2Faculty <strong>of</strong> Science at Charles' University, Department <strong>of</strong> Philosophy and History <strong>of</strong><br />
Science, Vinicna 7, Praha 2, 128 44, Czech Republic<br />
E-mail: arnost.l.sizling@seznam.cz<br />
Abstract<br />
Between 1993 and 2000, a sample <strong>of</strong> 646 individuals belonging to <strong>the</strong> T. cristatus<br />
group were collected and measured at sites in <strong>the</strong> Czech and Slovak Republics.<br />
Additionally, 26 individuals <strong>of</strong> T. carnifex from Italy and Slovenia, 8 <strong>of</strong> T. dobrogicus<br />
from Hungary and Rumania, and 6 <strong>of</strong> T. karelinifrom Turkey were collected for <strong>the</strong> purposes<br />
<strong>of</strong> comparison with <strong>the</strong> newts determined as T. carnifex and T. dobrogicus in <strong>the</strong><br />
Czech and Slovak Republics. All <strong>of</strong> <strong>the</strong>se individuals were measured with manual callipers<br />
under anaes<strong>the</strong>sia by <strong>the</strong> same person to <strong>the</strong> nearest 0.05 mm in <strong>the</strong> standardised<br />
manner.<br />
The quantities measured were W (weight), L (snout-vent-length), Led (length <strong>of</strong> tail),<br />
L.c. (length <strong>of</strong> head), Pa (average length <strong>of</strong> forelimbs), Pp (average length <strong>of</strong><br />
hindlimbs), LiE (interlimb distance), and Ltc (width <strong>of</strong> head). The colours <strong>of</strong> <strong>the</strong> body<br />
and <strong>the</strong> existence <strong>of</strong> <strong>the</strong> crest were also recorded. The hypo<strong>the</strong>sis that <strong>the</strong> Wolterstorff<br />
Index (Wl) - which is counted as 100*Pa/LiE - separates <strong>the</strong> crested newt group into<br />
single species has been tested. As expected, T. dobrogicus is <strong>the</strong> only species that Wl is<br />
able to discriminate. However, we received <strong>the</strong> most significant results using modified<br />
Wl calculated as 100*Pa/(LiE+6) for males and 100*Pa/(LiE+22) for females.<br />
Key words: news, Cristatus group, Wolterstorff Index<br />
Received 28 February 2002; accepted 10 May 2002
182 Biota "5/i-a, aooa ZAVADIL & SIZLING<br />
INTRODUCTION<br />
Until recently, Triturus cristatus newts<br />
were supposed to be <strong>the</strong> only representatives<br />
<strong>of</strong> <strong>the</strong> T. cristatus group in <strong>the</strong><br />
Czech Republic (Rocek 1992). In 1993,<br />
Triturus dobrogicus was morphologically<br />
determined in south Moravia at <strong>the</strong> confluence<br />
<strong>of</strong> <strong>the</strong> Morava and Dyje rivers;<br />
pro<strong>of</strong> in accordance with ELFO followed<br />
almost immediately (Zavadil et al. 1994).<br />
In 1997, newts showing features typical<br />
Table 1. Measured values.<br />
<strong>of</strong> T. carnifex were found near <strong>the</strong> town<br />
<strong>of</strong> Znojmo, at a site close to <strong>the</strong> border<br />
with Austria (Zavadil & Reiter unpubl.).<br />
Three years later, morphological analysis<br />
conducted by Pialek et al. (2000) proved<br />
that <strong>the</strong> observed population belongs to<br />
T. carnifex. ELFO analysis by Horak &<br />
Pialek (1999) determined <strong>the</strong> subpopulations<br />
as hybrids T. carnifex x T. cristatus<br />
and T. cristatus x T. dobrogicus.<br />
This evidence <strong>of</strong> morphological variability<br />
Short Name Fool Name Unit<br />
W Weight grams<br />
L snout-vent-length millimetres<br />
Led length <strong>of</strong> tail millimetres<br />
L.c.l length <strong>of</strong> head millimetres<br />
L.c. 2 length <strong>of</strong> head millimetres<br />
Pa average length <strong>of</strong> forelimbs millimetres<br />
Pp average length <strong>of</strong> hindlimbs millimetres<br />
LiE interlimb distance millimetres<br />
Ltc width <strong>of</strong> head millimetres<br />
Sp. oc. inter eyes distance millimetres<br />
Cr. max. cd. maximally width <strong>of</strong> tail millimetres<br />
Lc2<br />
Lc 1<br />
Figure 1. Measured values. See also Table 1.<br />
Led
ZAVADIL & SIZLING BJOLa 3/1-2, aooa 183<br />
motivated us to start research at multiple<br />
sites in <strong>the</strong> Czech and Slovak Republics.<br />
In this paper we try to answer <strong>the</strong> following<br />
questions, which can help resolve<br />
issues concerning <strong>the</strong> status <strong>of</strong> T. cristatus<br />
behind <strong>the</strong> Alps and T. dobrogicus at <strong>the</strong><br />
edge <strong>of</strong> its range:<br />
Are <strong>the</strong> Czech populations <strong>of</strong> T. carnifex<br />
and T. dobrogicus morphologically different<br />
from <strong>the</strong> Czech population <strong>of</strong> T.<br />
cristatus?<br />
Are <strong>the</strong> Czech populations <strong>of</strong> T. carnifex<br />
and T. dobrogicus (which are considered<br />
as marginal or isolated) morphologically<br />
different from <strong>the</strong> populations <strong>of</strong> T.<br />
carnifex and T. dobrogicus populations in<br />
core areas <strong>of</strong> <strong>the</strong>ir distributions?<br />
Is <strong>the</strong>re any geographic variability in body<br />
shape within <strong>the</strong> T. cristatus group?<br />
MATERIAL AND METHODS<br />
Between 1993 and 2000, 646 individuals<br />
belonging to <strong>the</strong> T. cristatus group were<br />
measured at sites in <strong>the</strong> Czech and Slovak<br />
Republics. For <strong>the</strong> purposes <strong>of</strong> comparison<br />
with <strong>the</strong> newts determined <strong>the</strong>re as<br />
T. carnifex and T. dobrogicus, an additional<br />
26 individuals <strong>of</strong> T. carnifex from<br />
Slovenia and Italy, 8 <strong>of</strong> T. dobrogicus<br />
from Hungary and Rumania, and 6 <strong>of</strong> T.<br />
karelinii from Turkey were examined.<br />
Since inaccuracy may be caused when<br />
measurements are taken by different persons<br />
or on animals conserved in alcohol<br />
(Arntzen & Wallis 1994), all <strong>the</strong> individuals<br />
were measured with manual callipers<br />
by <strong>the</strong> same person to <strong>the</strong> nearest 0.05<br />
mm in <strong>the</strong> standardised manner and<br />
under anaes<strong>the</strong>sia. The anaes<strong>the</strong>tic used<br />
was 0.8% water solution <strong>of</strong> phenoxyethanol<br />
(0.8 millilitre <strong>of</strong> phenoxyethanol<br />
in 1 litre <strong>of</strong> water).<br />
In accordance with a suggestion <strong>of</strong><br />
Klepsch (1994), measurements were<br />
taken <strong>of</strong> weight (W), snout-vent-length<br />
(L), length <strong>of</strong> tail (Led), length <strong>of</strong> head<br />
Figure 2. The localities; large rings show sites on which <strong>the</strong> large samples were collected.
184 Biota 3/1-2,2002 ZAVADIL & SIZLING<br />
(I.e.), average length <strong>of</strong> forelimbs (Pa),<br />
average length <strong>of</strong> hindlimbs (Pp), interlimb<br />
distance (LiE), width <strong>of</strong> head (Ltc),<br />
distance between eyelids (Sp. oc) and<br />
maximum width <strong>of</strong> tail (cr. max. cd.) (see<br />
Table 1, Figure 1). Fur<strong>the</strong>rmore, <strong>the</strong><br />
colouring <strong>of</strong> <strong>the</strong> body and <strong>the</strong> existence<br />
<strong>of</strong> <strong>the</strong> crest were recorded.<br />
During <strong>the</strong> first three years, <strong>the</strong> studied<br />
sites in <strong>the</strong> Czech Republic were chosen<br />
randomly. Later on, as we focused on differences<br />
between <strong>the</strong> T. cristatus and T.<br />
dobrogicus species, we concentrated on<br />
localities <strong>of</strong> T. cristatus remote from areas<br />
inhabited by T. dobrogicus and on localities<br />
situated at higher altitudes.<br />
Since 1997, we have focused on sites<br />
around <strong>the</strong> town <strong>of</strong> Znojmo, where <strong>the</strong> T.<br />
carnifex had been found; since 1998 we<br />
have examined sites in Slovakia situated<br />
on <strong>the</strong> border between <strong>the</strong> ranges <strong>of</strong> T.<br />
cristatus and T. dobrogicus (see Figure 2).<br />
We used <strong>the</strong> Analysis <strong>of</strong> Principal<br />
Component (PCA) for comparison <strong>of</strong><br />
samples from locations within <strong>the</strong> former<br />
Yugoslavia (Kalezic 1990) and <strong>the</strong><br />
Wolterstorff Index (Wl) for comparison<br />
with <strong>the</strong> results presented by Wolterstorff<br />
(1924), Arntzen & Wallis (1994) and<br />
Pialek et al. (2000).<br />
Since nei<strong>the</strong>r <strong>of</strong> <strong>the</strong> methods could satisfactorily<br />
show <strong>the</strong> existence <strong>of</strong> a morphometric<br />
difference between T. cristatus<br />
and T. carnifex, we decided to use <strong>the</strong><br />
Discriminant Analysis (DA) and <strong>the</strong> Cube<br />
Analysis (dividing <strong>the</strong> W-L-Lcd-L.c.-Pa-<br />
Pp-LiE-Ltc-Sp.oc.-cr.max.cd. space into<br />
three coherent areas consisting <strong>of</strong> bond<br />
cubes; see Appendix).<br />
However, <strong>the</strong> best results were reached<br />
by a modification <strong>of</strong> Wl, which we<br />
named Intensified Wolterstorff Index<br />
(i-WI; see below).<br />
RESULTS AND DISCUSSION<br />
Multivariate analyses<br />
Three accomplished multivariate analyses<br />
(PCA, Discriminant Analysis and Cube<br />
Analysis - see Appendix) were used to<br />
measure morphological and geographical<br />
differences between studied species. The<br />
results <strong>of</strong> PCA are concordant with <strong>the</strong><br />
works <strong>of</strong> Kalezic et al. (1990) and Pialek<br />
& Zavadil (1997); however, <strong>the</strong> clusters<br />
shown in our study were larger and consequently<br />
overlapped. To obtain <strong>the</strong> best<br />
result using Discriminant Analysis we<br />
reduced measured variables so that <strong>the</strong>y<br />
were linearly independent (we used all<br />
variables except W and L) and used a<br />
quadratic model because <strong>of</strong> inequality <strong>of</strong><br />
covariance matrices and transformed<br />
axes to normalize its distribution and<br />
eliminate allometry. However, even when<br />
all assumptions <strong>of</strong> DA were met as accurately<br />
as possible, we did not obtain significantly<br />
better results (measured in percentage<br />
<strong>of</strong> successful determination) than<br />
we obtained using only Pa and LiE variables.<br />
This result suggests that <strong>the</strong>se two<br />
variables are adequate, but in <strong>the</strong> end we<br />
constructed and carried out <strong>the</strong> Cube<br />
Analysis to guarantee this result.<br />
Therefore, we concentrate on <strong>the</strong>se two<br />
variables in <strong>the</strong> following text.<br />
Wl analysis<br />
Wolterstorff Index (Wl) analysis is a classical<br />
method used to identify species<br />
belonging to <strong>the</strong> T. cristatus group. It is<br />
defined as 100*Pa/LiE. Though <strong>the</strong> Wl is<br />
very helpful for <strong>the</strong> determination <strong>of</strong> T.<br />
dobrogicus, it is not satisfactory for distinguishing<br />
between T. cristatus and T.<br />
carnifex (Kalezic et al. 1990, Arntzen &<br />
Wallis 1994, 1999). The problem is that<br />
<strong>the</strong> Wl decreases with growing body<br />
length (see Figure 3 where <strong>the</strong> Wl corresponds<br />
to <strong>the</strong> slope <strong>of</strong> <strong>the</strong> dotted lines).<br />
When studying <strong>the</strong> Pa-LiE space (see<br />
Figure 3), we realised that <strong>the</strong> decrease in<br />
observed Wl was related to ineffective<br />
distinction between <strong>the</strong> two species.<br />
There is linearity between Pa and LiE, but<br />
<strong>the</strong> borderlines/regression lines do not<br />
attain zero. This could be caused by a dif-
ZAVADIL & SIZLING 3/1-2, 2OO2 185<br />
Figure 3. Males-top, Females-bottom. There is a difference between Wl dotted lines<br />
and i-WI <strong>full</strong> lines in Figs, a, b: rings correspond to T. carnifex, filled circles to T. cristatus,<br />
triangles to T. dobrogicus and sharps to T. karelinii; sharps inside <strong>the</strong> circles mark<br />
external individuals <strong>of</strong> T. ca. (I, SLO) and diamonds external individuals <strong>of</strong> T. do. (H,<br />
RO).<br />
[interlimb Distance (LIE) in mms<br />
<strong>full</strong> lines (i-WI) does not attain zero<br />
dotted lines (Wl) attain zero<br />
ferent rate <strong>of</strong> growth in <strong>the</strong> larvae stage.<br />
It is shown on Figure 3 where <strong>the</strong> dotted<br />
lines correspond to Wl and <strong>the</strong> coloured<br />
lines correspond to <strong>the</strong> intensified Wl,<br />
which is calculated as 100*Pa/(LiE+22)<br />
for females and 100*Pa/(LiE+6) for males<br />
(see Appendix). The classical way to eliminate<br />
allometry helping <strong>the</strong> logarithm<br />
Interlimb Distance (LiE) in mms<br />
function appears to be less powerful.<br />
Box and Whisker plot <strong>of</strong> <strong>the</strong> i-WI is<br />
shown in Figure 4 and <strong>the</strong> effectiveness<br />
<strong>of</strong> discrimination in Table 2.<br />
If <strong>the</strong> i-WI should turn out to increase<br />
with growing length measures, it is probably<br />
caused by <strong>the</strong> simplification <strong>of</strong> its<br />
original form, which is ATN(Pa/(LiE+22))
186 Biota 3/i-a, 2002 ZAVADIL & SIZLING<br />
Figure 4. Males-top, Females-bottom. Box and whisker plot <strong>of</strong> i-WI for different localities.<br />
The size <strong>of</strong> sample "n" is for males 12 for site 1041, 21 for1161, 13for1171,31<br />
for 1181, 15 for 1211, 12 for 1291, 6 for 1351, 8 for 1401, 22 for 1411, 4 for 1421, 6<br />
for 1451, 15 for 1491, 6 for 2021, 7 for 2051, 7 for 2061, 11 for 2071, 1 for 3011, 1<br />
for 4011, 11 for 5011, 2 for 6021 and 4 for 7011. For females <strong>the</strong> size <strong>of</strong> sample "n"<br />
is 4 for 1041, 17 for 1171, 22 for 1181, 14 for 1211, 10 for 1291, 9 for 1351, 8 for<br />
1391, 4 for 1401, 30 for 1411, 9 for 1451, 9 for 1481, 5 for 1491, 9 for 2021, 9 for<br />
2061, 6 for 2071, 3 for 3011, 3 for 4011, 10 for 5011, 2 for 6021 and 2 for 7011.<br />
cu<br />
1041 1171 1211 1351 1411 1451 2021 2061 3011 5011 7011<br />
1161 1181 1291 1401 1421 1491 2051 2071 4011 6021<br />
1041 1181 1291 1391 1411 1481 2021 2071 4011 6021<br />
1171 1211 1351 1401 1451 1491 2061 3011 5011 7011
ZAVADIL & SIZLING Biota 3 i-2,2002 187<br />
Table 2. Effectiveness <strong>of</strong> i-WI Analysis.<br />
MALES; i-WI = 100*Pa/(LiE+6) % <strong>of</strong> Successful<br />
Determination<br />
Species T. ca. T. cr. T.do.<br />
T. ca. 45 9 0<br />
83<br />
T. cr. 7 135 0<br />
95<br />
T.do. 0 4 68<br />
94<br />
FEMALES; i-WI = 100*Pa/(LiE+22)<br />
T. ca. T. cr. T.do.<br />
T. ca. 35 7 0<br />
83<br />
T. cr. 1 154 8<br />
94<br />
T.do. 0 0 67<br />
100<br />
Table 3. Threshold values for Wl and i-WI according to different authors; many <strong>of</strong> <strong>the</strong>se<br />
were taken from graphs<br />
WI - Threshold value<br />
Males<br />
Females<br />
T. karelinii<br />
T. carnifex<br />
T. cristatus<br />
T. dobrogicus<br />
T. karelinii<br />
T. carnifex<br />
T. cristatus<br />
T. dobrogicus<br />
Our Study i-WI<br />
from to<br />
55.5<br />
45.5<br />
32<br />
35.3<br />
28.7<br />
22<br />
63<br />
55.5<br />
45.5<br />
40<br />
35.3<br />
28.7<br />
Our Study Wl<br />
from to<br />
70 75<br />
60 75<br />
50 69<br />
38 55<br />
60 65<br />
50 65<br />
32 61<br />
30 ' 45<br />
for females and ATN(Pa/(LiE+6)) for<br />
males, where ATM is <strong>the</strong> ma<strong>the</strong>matical<br />
function arc tangent. However, <strong>the</strong>re is<br />
good statistical evidence that <strong>the</strong> i-WI<br />
increases with growing width sizes (Pa,<br />
Pp, Ltc, P < 0.05). All results remained<br />
unchanged when <strong>the</strong> data for subadult<br />
individuals had been added.<br />
The thresholds <strong>of</strong> Wl and i-WI can be<br />
found in Table 3. The evidence that <strong>the</strong><br />
Wl is higher for males than for females is<br />
much stronger for samples collected<br />
within single sites than for samples collected<br />
on more sites (see Figure 4).<br />
Geographical variability<br />
We have no evidence for morphological<br />
difference between <strong>the</strong> Czech population<br />
<strong>of</strong> T. carnifex and populations <strong>of</strong> T.<br />
Wolterstorff 1924<br />
From to<br />
69 82<br />
63 73<br />
59.8 65<br />
45 52<br />
67.5 72<br />
52 64<br />
49 54<br />
34 45<br />
Kalezicetal.1990 Arntzen & Walis 1994<br />
from to from to<br />
69 81 57 67<br />
63 73 49 64<br />
55 65 44 62<br />
45 51 35 46<br />
61 72 57 67<br />
54 65 49 64<br />
45 54 44 62<br />
34 45 35 46<br />
carnifex south <strong>of</strong> <strong>the</strong> Alps and T. dobrogicus<br />
in <strong>the</strong> Czech and Slovak Republics<br />
and <strong>the</strong> same species abroad. This is<br />
because we have no evidence that for <strong>the</strong><br />
same species, <strong>the</strong> difference between <strong>the</strong><br />
Czech population and populations<br />
abroad is bigger than <strong>the</strong> difference<br />
among Czech populations (see Figure 4).<br />
Although <strong>the</strong>re are sites where <strong>the</strong> same<br />
species <strong>of</strong> <strong>the</strong> T. carnifex group show a<br />
statistically different body shape (see<br />
Figure 4), it would not be correct to speak<br />
about geographical variability. The reason<br />
is that no correspondence was found<br />
between <strong>the</strong> shape (i-WI and <strong>the</strong> results<br />
<strong>of</strong> <strong>the</strong> Discriminant Analyses) and any<br />
geographical attribute (altitude, <strong>the</strong> distance<br />
between sites or any direction, as,<br />
for example, East, West, SE, etc.).
188 Biota 3/i-a, 2002 ZAVADIL & SIZLING<br />
A sound hypo<strong>the</strong>sis seems to be that <strong>the</strong><br />
morphological differences between particular<br />
sites can be caused by different<br />
densities <strong>of</strong> larvae predators (Schmidt &<br />
Van Buskirk2001).<br />
Appendix<br />
The Analysis <strong>of</strong> Principal Components<br />
(PCA) transforms <strong>the</strong> axes <strong>of</strong> <strong>the</strong> examined<br />
orthogonal space in order to show<br />
clusters <strong>of</strong> <strong>the</strong> studied samples. If it is not<br />
possible to see any clusters, <strong>the</strong>re is no<br />
evidence for <strong>the</strong>ir absence. The PCA was<br />
used for data analyses in <strong>the</strong> works <strong>of</strong><br />
Kalezic et al. (1990) and Pialek & Zavadil<br />
(1997).<br />
The Discriminant Analysis calculates a<br />
number D for a case (a single newt), and<br />
for each group <strong>of</strong> a studied sample (TV<br />
cristatus, T. carnifex, T. dobrogicus). The<br />
group with <strong>the</strong> lowest D is assumed to be<br />
<strong>the</strong> group into which <strong>the</strong> case (newt)<br />
belongs.<br />
The purpose <strong>of</strong> <strong>the</strong> Cube Analysis is to<br />
divide morphometric space into two continuous<br />
parts, which contain ei<strong>the</strong>r T.<br />
cristatus or T. carnifex individuals. In <strong>the</strong><br />
case <strong>of</strong> ten-dimensional space it would be<br />
very difficult, so in <strong>the</strong> beginning we<br />
reduced <strong>the</strong> data with <strong>the</strong> help <strong>of</strong> PCA<br />
(<strong>the</strong> axes with <strong>the</strong> lowest variability are<br />
eliminated). After data reduction, we<br />
divided <strong>the</strong> reduced space into optimal<br />
size cubes, in <strong>the</strong> same way that biologists<br />
divide maps into quadrants for<br />
quadrant mapping. The optimal size <strong>of</strong><br />
<strong>the</strong> cubes was taken empirically by testing<br />
various sizes. The cubes create three<br />
continuous areas: <strong>the</strong> first one contains<br />
only T. carnifex, <strong>the</strong> second one only T.<br />
cristatus, and <strong>the</strong> third one consists <strong>of</strong><br />
cubes in which both species could be<br />
found. This algorithm was applied subsequently<br />
on <strong>the</strong> third area.<br />
Wl and i-WI analyses. In Figure 1 in<br />
Arntzen & Wallis (1994) <strong>the</strong> T. cristatus<br />
and T. marmoratus are plotted in Pa-LiE<br />
state space. Making <strong>the</strong> best line for distinguishing<br />
between <strong>the</strong> two species<br />
(error <strong>of</strong> determination in percent is <strong>the</strong><br />
lowest) and extrapolating it, we realized<br />
that <strong>the</strong> line does not attain zero and so it<br />
can not represent a threshold for <strong>the</strong> Wl.<br />
This is because <strong>the</strong> formula Wl =<br />
1QO*Pa/LiE is <strong>the</strong> o<strong>the</strong>r form for a line<br />
which attains zero. The form <strong>of</strong> this line<br />
can be written as Pa = (WI/100)*LiE.<br />
This phenomenon is also clearly apparent<br />
in our data (see Figure 3). It means that<br />
threshold lines in <strong>the</strong> Pa-LiE state space<br />
do not attain zero, and <strong>the</strong> common form<br />
<strong>of</strong> <strong>the</strong>se lines must be Pa =<br />
(i-WI/100)*LiE+b, where b represents<br />
<strong>the</strong> point <strong>of</strong> intersection with <strong>the</strong> Pa axis.<br />
This equation can be written as i-WI =<br />
100*Pa/(LiE+c) where c is computed for<br />
males and females separately (b =<br />
c*[i-WI/100]). The number i-WI we call<br />
intensified Wolterstorff index.<br />
Acknowledgements<br />
Special thanks to J. Pialek, D. Weber-Obdrzalkova, Jan Sula and Lukas Kratochvfl for<br />
<strong>the</strong>ir assistance and to P. Belansky, D. Cogalniceanu, P. Dolezal, A. Horak, M. Kaftan, J.<br />
Kautman, S. Koukal, A/I. Macholan, K. Poboljsaj, Puky M., A. Reitr, K. Rozfnek, R.<br />
Rozinek, A. Ruxova, R. Zajfcek and J. Zima for <strong>the</strong>ir help during <strong>the</strong> collection <strong>of</strong> <strong>the</strong><br />
data sample.<br />
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ZUFFI, CENTILUI, RAZZETTI & SCALI Biota 3/1-2. 2002 191<br />
Transition-hybridization areas in<br />
parapatric species <strong>of</strong> Vipera aspis<br />
group from nor<strong>the</strong>rn Italy<br />
Marco A.L ZUFFI1*P Augusto GENTILLI2,<br />
Edoardo RAZZETTI2 & Stefano SCALP<br />
'"Corresponding author: Museo di Storia Naturale e del Territorio, University <strong>of</strong> Pisa, via<br />
Roma 79, 1-56011 Calci (Pisa)-Italy<br />
E-mail: marcoz@museo.unipi.it<br />
2Dip. Biologia Animale, University <strong>of</strong> Pavia, p.za Botta 9, 27100 Pavia, Italy<br />
3Museo Civico di Storia Naturale, c.so Venezia 55, 20121 Milano-ltaly<br />
Abstract<br />
There is evidence <strong>of</strong> some contact between <strong>the</strong> parapatric species <strong>of</strong> Vipera aspis, Italian<br />
V. aspis and Vipera atra Meisner, 1820 along a North-South oriented sector, from about<br />
8°45' to 9° 20' Longitude East. V. atra from north-western Italy is characterized by a<br />
high number <strong>of</strong> ventral scales and dorsal bars, and by a distinct morphological pattern<br />
<strong>of</strong> <strong>the</strong> hemipenes that has elongated lobes, mostly covered by short and small spines,<br />
while V. aspis has a relatively lower number <strong>of</strong> ventral scales and dorsal bars, and<br />
hemipenes that show large lobes with <strong>the</strong> upper part completely covered by a calyculate<br />
area. The intermediate specimens appear to be more similar to V.aspis than to V.<br />
atra.<br />
Key words: Vipera aspis (species group), nor<strong>the</strong>rn Italy, contact zones.<br />
Received: 3 October 2001; accepted 12 April 2002
192 Biota 3/1-2,2002 ZUFFI, GENTILLI, RAZZETTI & SCALI<br />
INTRODUCTION<br />
Paleontological and molecular biology<br />
data indicate that <strong>the</strong> small vipers <strong>of</strong> <strong>the</strong><br />
western Palaearctic region, including <strong>the</strong><br />
Sand viper, Vipera ammodytes, and <strong>the</strong><br />
Asp viper, Vipera aspis, represent a complex<br />
<strong>of</strong> phylogenetically closely related<br />
taxa (Herrmann & Joger 1997, Lenk et al.<br />
2001).<br />
The Asp viper, with regards to colour<br />
morphs and dorsal markings, is one <strong>of</strong> <strong>the</strong><br />
most variable snakes <strong>of</strong> <strong>the</strong> Palaearctic<br />
region (Physalix 1968, Brodmann 1987).<br />
Among <strong>the</strong> several subspecies which<br />
have been described in <strong>the</strong> past (cf. Saint<br />
Girons 1997), only atra, frandsciredi,<br />
hugyi, zinnikeri and <strong>the</strong> nominal aspis<br />
have been accepted (Zuffi & Bonnet<br />
1999) (Figure 1). More recently, some <strong>of</strong><br />
<strong>the</strong>se taxa have been recognised as good<br />
species (Zuffi 2002) and <strong>the</strong>y form part <strong>of</strong><br />
interest <strong>of</strong> this contribution. Inter and<br />
intra-population analysis <strong>of</strong> life history<br />
traits represents one <strong>of</strong> <strong>the</strong> most fascinating<br />
approaches towards <strong>the</strong> description<br />
and <strong>the</strong> comprehension <strong>of</strong> phenotypic<br />
plasticity (Seigel & Fitch 1985, Bonnet &<br />
Naulleau 1996, Shine 2000, Luiselli &<br />
Zuffi 2002). The high morphological variability<br />
<strong>of</strong> <strong>the</strong> Asp viper, and <strong>the</strong> wide<br />
range <strong>of</strong> habitats in which this species<br />
lives (Saint Girons 1997), <strong>of</strong>fer an opportunity<br />
for multidisciplinary studies in evolutionary<br />
biology.<br />
The analysis <strong>of</strong> <strong>the</strong> number <strong>of</strong> ventral and<br />
caudal scales <strong>of</strong> nor<strong>the</strong>rn Italian Vipera<br />
aspis shows, within a North-South oriented<br />
sector (from about 8°45 to 9°20'<br />
Longitude East), that V. aspis specimens<br />
display intermediate characters between<br />
those <strong>of</strong> V. atra and <strong>of</strong> Italian V. aspis.<br />
This geographic sector extends from <strong>the</strong><br />
western and eastern sides <strong>of</strong> <strong>the</strong> Ticino<br />
river, from <strong>the</strong> Maggiore lake to <strong>the</strong> confluence<br />
with <strong>the</strong> Po river; from <strong>the</strong> Ticino-<br />
Po confluence southwards to <strong>the</strong> begin-<br />
Figure 1. Present distribution <strong>of</strong> European Vipera aspis subspecies.<br />
40° lat N<br />
frandsciredi<br />
500 Km
ZUFFI, GENTILLI, RAZZETTI & SCALI Biota 3/1-2,2002 193<br />
Figure 2. Contact zone between Vipera atra and Italian V. aspis. Circles = atra; Boxes =<br />
Italian aspis; Crosses = intermediate individuals<br />
ning <strong>of</strong> <strong>the</strong> Apennines; and along a relatively<br />
large area that connects, along a<br />
virtual line, <strong>the</strong> towns <strong>of</strong> Voghera,<br />
Tortona, Novi Ligure, and Genoa.<br />
This contribution presents <strong>the</strong> first available<br />
morphological data on V. aspis populations<br />
<strong>of</strong> two distinct species, i.e. Vipera<br />
atra and Italian Vipera aspis from northwestern<br />
Italy, along a western-eastern<br />
transect, passing through <strong>the</strong> border<br />
between <strong>the</strong> species (Figure 2). We present<br />
data documenting <strong>the</strong> parapatric<br />
occurrence <strong>of</strong> two species <strong>of</strong> V. aspis<br />
along with a limited number <strong>of</strong> intermediates.<br />
MATERIALS AND METHODS<br />
173 vipers (55 V. atra. 104 V. aspis, 14<br />
intermediate specimens), (Zuffi & Bonnet<br />
1999, Zuffi 2002) were studied for <strong>the</strong><br />
following characteristics: snout-vent<br />
length (SVL), tail length (tL), total length<br />
(TL), ventral scales (VS), subcaudal scales<br />
(CS), total scales (TOTS), bars or blotches<br />
or spots on <strong>the</strong> dorsal left side (BARS).<br />
Because <strong>of</strong> strong sexual dimorphism,<br />
analyses were considered for each sex<br />
separately. 22 vipers were analysed for<br />
hemipenial morphology as described in<br />
Zuffi (2002). Moreover, 35 individuals<br />
from <strong>the</strong> Enrica Calabresi herpetological<br />
collection (University <strong>of</strong> Florence) were<br />
examined for differences in skull morphology.<br />
Calabresi (1924) pointed out<br />
that <strong>the</strong> shape <strong>of</strong> parietal and basisphenoid<br />
bones are quite characteristic in<br />
Italian V. aspis subspecies. We used differences<br />
in <strong>the</strong> parietal and basisphenoid<br />
morphology to evaluate <strong>the</strong> skulls.<br />
Parietal bone shows a different pattern<br />
for each <strong>of</strong> <strong>the</strong> two species: i) in V. atra<br />
<strong>the</strong> parietal does not show any bony<br />
crest; ii) in V. aspis <strong>of</strong> Italy lateral bony<br />
crests are typically present, and <strong>the</strong>y are<br />
convergent posteriorly contacting each<br />
o<strong>the</strong>r in "V" shaped crests. Basisphenoid<br />
bones show different patterns in each<br />
species: i) in V. atra <strong>the</strong> proximal portion<br />
<strong>of</strong> <strong>the</strong> bone, at <strong>the</strong> level <strong>of</strong> <strong>the</strong> postorbitals,<br />
is a romboid shaped crest that con-
194 Biota 3/1-2,2002 ZUFFI, GENTILLI, RAZZETTI & SCALI<br />
tinues posteriorly as a longitudinal narrow<br />
crest, forming a single process, ending at<br />
<strong>the</strong> basioccipital suture; in ii) V. aspis<br />
such a ventral pattern is generally absent;<br />
<strong>the</strong> proximal part <strong>of</strong> <strong>the</strong> basisphenoid in<br />
V. aspis possesses a narrow longitudinal<br />
crest that continues, in <strong>the</strong> distal part <strong>of</strong><br />
<strong>the</strong> basisphenoid bone, as a small romboid<br />
shaped crest. Body size parameters<br />
were analysed to evaluate <strong>the</strong> variation<br />
along <strong>the</strong> transect, without regard to taxonomic<br />
status <strong>of</strong> each specimen. After<br />
this we used additional characteristics to<br />
assign taxonomic status as V. atra or V.<br />
aspis or as intermediate. The original data<br />
set was log-transformed to avoid <strong>the</strong><br />
effect <strong>of</strong> allometry; data were <strong>the</strong>n tested<br />
for normality. We used SPSS 8.0 PC to<br />
analyse our data. All tests were two tailed<br />
and we chose P = 0.05 as our level <strong>of</strong> significance.<br />
RESULTS<br />
Body size features<br />
There was no significant difference in SVL<br />
or in TL between V. atra and V. aspis<br />
males and females. Vipera atra males differed<br />
in CS and TOTS when compared to<br />
females (males CS: 43.7 ± 3.1, N=23,<br />
males TOTS: 198 ± 5, N = 23; vs. females<br />
CS: 37.6 ± 3.4, N = 28, females TOTS:<br />
192.3 ± 5, N = 27; Student t test = 6.569,<br />
49 df, P < 0.001, and t = 4.001, 48 df, P<br />
< 0.0001). Vipera aspis males differed<br />
significantly from females in VS (146.6 ±<br />
3.4, N = 35, vs. 150.4+ 3.7, N = 66,<br />
Student t test = -4.813, 103 df, P =<br />
0.0001), CS (42 ± 2.5, N = 32 vs. 33.9 ±<br />
2.3, N = 65, Student t test = 15.894, 99<br />
df, P = 0.0001, p), TOTS (188.4 ± 3.9, N<br />
= 31, vs. 184.3 ± 4.1, N = 63, Student t<br />
test = 4.726, 96 df, P = 0.0001), and<br />
BARS (40.7 ± 2.1, N = 24, vs. 43.7 ± 3.8,<br />
N = 36, Student t test = -2.435, 62 df, P<br />
= 0.018) respectively. Vipera aspis males<br />
<strong>of</strong> <strong>the</strong> intermediate form differed from<br />
females in CS (44 ± 2.5, N = 5, vs. 36.1 ±<br />
2.1, N = 8, Student t test = 3.8, 15 df, P<br />
= 0.002).<br />
Vipera atra males possessed a significantly<br />
higher number <strong>of</strong> ventrals, subcaudals,<br />
totals and dorsal bars ei<strong>the</strong>r <strong>of</strong> Italian V.<br />
aspis specimens (VS, TOTS, BARS, P =<br />
0.0001; CS, P = 0.031; One-Way<br />
ANOVA, LSD multirange test), or those<br />
<strong>of</strong> intermediate specimens (VS, TOTS,<br />
BARS, P = 0.0001). On <strong>the</strong> o<strong>the</strong>r hand,<br />
V. aspis <strong>of</strong> Italy and <strong>the</strong> individuals <strong>of</strong> <strong>the</strong><br />
intermediate form were close to each<br />
o<strong>the</strong>r for VS (P = 0.095); CS (P = 0.786);<br />
TOTS (P = 0.259) and BARS (P = 0.22).<br />
Vipera atra females had a significantly<br />
higher number <strong>of</strong> ventrals, subcaudals,<br />
totals and dorsal bars than V. aspis specimens<br />
(VS, CS, TOTS.BARS, P = 0.0001),<br />
as well as <strong>the</strong> number <strong>of</strong> ventrals, totals<br />
and dorsal bars <strong>of</strong> <strong>the</strong> intermediate specimens<br />
(VS, P = 0.004; TOTS, P = 0.002;<br />
BARS, P = 0.016).<br />
Skull morphology<br />
We did not observe any appreciable sexual<br />
differences in <strong>the</strong> skull morphology <strong>of</strong><br />
ei<strong>the</strong>r <strong>the</strong> parietal or basisphenoid bones.<br />
In <strong>the</strong> study area we found that <strong>the</strong> V. a.<br />
atra pattern <strong>of</strong> <strong>the</strong> parietal bone occurs in<br />
87.5% <strong>of</strong> <strong>the</strong> aspis classified individuals<br />
(N = 8); <strong>the</strong> V. aspis pattern occurs in<br />
76% <strong>of</strong> V. aspis classified individuals (N =<br />
21). The pattern <strong>of</strong> <strong>the</strong> intermediate<br />
specimens (N = 6) was similar to V. atra<br />
in 33.3% <strong>of</strong> <strong>the</strong> cases, and to V. aspis in<br />
66.6% <strong>of</strong> <strong>the</strong> cases.<br />
Hemipenial morphology<br />
Hemipenes <strong>of</strong> French V. aspis show well<br />
separated lobes, relatively longer than <strong>the</strong><br />
basal segment; <strong>the</strong> basal segment possesses<br />
large basal spines reaching <strong>the</strong><br />
bifurcation <strong>of</strong> <strong>the</strong> sperm groove and<br />
medium to small sized spines up to last<br />
third to fourth <strong>of</strong> <strong>the</strong> lobe with a relatively<br />
large calyculate area (Domergue 1962:<br />
98, Fig. 10, Case 1968: 98, Fig. 3, Joger<br />
et al., 1997, Zuffi 2002). In comparison,<br />
V. atra hemipenes have well separated
ZUFFI, GENTILLI, RAZZETTI & SCALI 3/1-2 195<br />
lobes that are relatively longer than <strong>the</strong><br />
basis, with large basal spines and many<br />
small spines covering both asulcated and<br />
sulcated sides up to <strong>the</strong> apex, with a very<br />
reduced calyculate area. Italiann Vipera<br />
aspis shows strong similarities to French<br />
V. aspis: it has similar basal segment and<br />
spines, medium to small spines and a relatively<br />
well defined calyculate area.<br />
However, its hemipenes are, on average,<br />
larger than those <strong>of</strong> <strong>the</strong> nominal and <strong>the</strong><br />
o<strong>the</strong>r subspecies. The only one intermediate<br />
specimen examined for this characteristic<br />
shows an intermediate hemipenial<br />
morphology, sharing <strong>the</strong> features <strong>of</strong> both<br />
V. atra and V. aspis <strong>of</strong> Italy.<br />
DISCUSSION<br />
Vipera aspis Is highly polymorphic regarding<br />
dorsal patterns <strong>of</strong> colouration and<br />
markings (Brodmann 1987, Zuffi &<br />
Bonnet 1999). Our investigation indicates<br />
that V. atra and Italian V. aspis are distin-<br />
guishable by <strong>the</strong> analysis <strong>of</strong> ventrals, dorsal<br />
bars, morphology <strong>of</strong> skull bones, and<br />
hemipenial morphology. It is only in <strong>the</strong><br />
contact zone that some <strong>of</strong> <strong>the</strong>m are individuals<br />
showing an intermediate pattern,<br />
more like V. aspis <strong>of</strong> Italy.<br />
Based on <strong>the</strong> fact that few individuals are<br />
intermediates in morphology, our results<br />
suggest some restriction in gene flow<br />
between V. atra and Italian V. aspis.<br />
Previous works seem to indicate a strong<br />
genetic separation between <strong>the</strong>se two<br />
forms (Pozio 1980) as well as at a morphological<br />
level (Zuffi 2002) as suggested<br />
by <strong>the</strong> marked difference <strong>of</strong> hemipene<br />
morphology between <strong>the</strong>se taxa. Genetic<br />
and inter-fertility studies have not been<br />
done yet but will provide additional evidence<br />
to help determine <strong>the</strong> systematic<br />
status <strong>of</strong> <strong>the</strong>se two taxa.<br />
REFERENCES<br />
BONNET, X. & NAULLEAU, G. 1996: Catchability in snakes: consequences for estimates <strong>of</strong><br />
breeding frequency. Can. J. Zool. 74: 233-239.<br />
BRODMANN, P. 1987: Die Giftschlangen Europas und die Gattung Vipera in Afrika und<br />
Asien. Kummerly-Frey.<br />
CALABRESI, E. 1924: Ricerche sulle variazioni della Vipera aspis Auct. in Italia. Boll. 1st. Zool.<br />
R. Univ. Roma, 2: 78-127.<br />
DOMERGUE, Ch. A. 1962: Observations sur le penis des Ophidiens. Bull. Soc. Sci. nat phys.<br />
Maroc42:87-105.<br />
GASC, J.-P. 1968: Morphologic des hemipenis chez Vipera ursinii ursinii (Bonaparte) et discussion<br />
biogeograpique sur la repartition des especes du genre Vipera en Europe<br />
occidentale. Bull. Mus. Nat. Hist. Nat. 40: 95-101.<br />
HERRMANN, H.-W. & JOGER, U., 1997: Evolution <strong>of</strong> viperine snakes. In: Thorpe, R.S.,<br />
Wuster, W. & Malhotra, A. Eds. Venomous Snakes: Ecology, Evolution and Snake<br />
Bite. Symp. zool. Soc. London 70: 43-61.<br />
JOGER, U., LENK, P., BARAN, I., BOHME, W., ZIEGLER, T, HEIDRICH, P. & WINK, M. 1997:<br />
The phylogenetic position <strong>of</strong> Vipera barani and V. nikolskii within <strong>the</strong> Vipera<br />
berus complex. In: Bohme, W., Bisch<strong>of</strong>f, W. & Ziegler, T. (eds). Herpetologia<br />
Bonnensis. Bonn (<strong>SEH</strong>), Germany: 185-194.<br />
LENK, P., KALYABINA, S., WINK, M. & JOGER, U. 2001: Evolutionary relationships among<br />
<strong>the</strong> true vipers (Reptilia: Viperidae) inferred from Mitochondria! DNA sequences.<br />
Molecular Phylogenetics Evolution 19: 94-104.
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LUISELLI, L. & ZUFFI, M.A.L. 2002: Female life-history traits <strong>of</strong> <strong>the</strong> aspic viper (Vipera aspis)<br />
and sand viper (V. ammodytes) from <strong>the</strong> Mediterranean region. In: Schuett<br />
G.W., Hoggren M., Douglas M.E. & Greene H.W. (eds.). Biology <strong>of</strong> Vipers.<br />
CPG/Biological Sciences Press, Carmel, Indiana: 279-284.<br />
PHISALIX, M. 1968: La livree des viperes de France. Bull. Mus. Nat. Hist. Nat., Paris 4: 661-<br />
676.<br />
POZIO, E. 1980: Contributo alia sistematica di Vipera aspis (L.) mediante analisi elettr<strong>of</strong>oretica<br />
delle proteine contenute nel veleno. Natura. Soc. ital. Sci. nat., Museo civ.<br />
Stor. nat. e Acquario civ., Milano, 71: 28-34.<br />
SAINT GIRONS, H., 1997: Vipera aspis (Linnaeus, 1758). In: Case, J.P., Cabela, A.,<br />
Crnobrnja-lsailovic, J., Dolmen, D., Grossenbacher, K.r Haffner, P., Lescure, J.,<br />
Martens, H., Martinez Rica, J.P., Maurin, H., Oliveira, M.E., S<strong>of</strong>ianidou, T.S.,<br />
Veith, M. & Zuiderwijk, A. (eds). Atlas <strong>of</strong> Amphibians and Reptiles in Europe.<br />
Societas Europaea Herpetologica and Museum National d'Histoire Naturelle<br />
(IEGP/SPN), Paris: 386-387.<br />
SHINE, R. 2000: Vertebral number in male and female snakes: <strong>the</strong> roles <strong>of</strong> natural, sexual<br />
and fecundity selection. J. Evol. Biol. 13: 49-86.<br />
SEIGEL, R.A., & FITCH, S. 1985: Annual variation in reproduction in snakes in a fluctuating<br />
environment. Journal <strong>of</strong> Animal Ecology 54: 497-505.<br />
ZUFFI, M.A.L. 2002: A critique <strong>of</strong> <strong>the</strong> systematic position <strong>of</strong> <strong>the</strong> asp viper subspecies [Vipera<br />
aspis aspis (Linne, 1758), Vipera aspis atra Meisner, 1820, Vipera aspis francisciredi<br />
Laurenti, 1768, Vipera aspis hugyi Schinz, 1833, Vipera aspis zinnikeri<br />
Kramer, 1958]. Amphibia-Reptilia 23: 191-213.<br />
ZUFFI, M.A.L. & BONNET, X. 1999: Italian subspecies <strong>of</strong> <strong>the</strong> asp viper, Vipera aspis: patterns<br />
<strong>of</strong> variability and distribution. Ital. Journal <strong>of</strong> Zoology 66: 87-95.
NOVE KNJIGE/BOOK REVIEWS<br />
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(eds.)2001:<br />
Atlas zur Verbreitung und Okologie der<br />
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880 pages.<br />
Hardback, ISBN 3-85457-586-6<br />
Copies can be ordered by e-mail:<br />
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This massive <strong>book</strong>, measuring 21 x 27 cm<br />
and weighing much more than 2 kg,<br />
reflects <strong>the</strong> efforts <strong>of</strong> a team <strong>of</strong> three<br />
main researchers at <strong>the</strong> Natural History<br />
Museum <strong>of</strong> Vienna who directed <strong>the</strong><br />
fieldwork <strong>of</strong> more than 400 field workers,<br />
edited <strong>the</strong> copy <strong>of</strong> <strong>the</strong> 12 authors who<br />
wrote particular chapters (18), and syn<strong>the</strong>sized<br />
all <strong>the</strong> cartographic information.<br />
The most substantial part <strong>of</strong> <strong>the</strong> atlas,<br />
pages 164-610, includes accounts <strong>of</strong> 37<br />
species (21 amphibian and 16 reptile<br />
species), from Salamandra atra (p. 164-<br />
174) to Vipera ursinii (605-610). On<br />
average, about 12 pages is devoted to<br />
each species.<br />
The result <strong>of</strong> <strong>the</strong>se intensive labours is an<br />
extremely handsome <strong>book</strong>, illustrated<br />
with beautiful colour photographs (157!)<br />
<strong>of</strong> each species and <strong>the</strong>ir habitats, numerous<br />
graphs and tables, and, <strong>of</strong> course,<br />
innumerable maps that include not only<br />
details <strong>of</strong> species distribution in Austria,<br />
but also topography. For each species<br />
extensive ecological and phenological<br />
data is shown in various graphs.<br />
Moreover, vertical distribution and sympatric<br />
species are also given.<br />
An unusual, but very useful, feature is<br />
that this Atlas contains some special<br />
chapters, such as an identification key for<br />
groups <strong>of</strong> reptiles and amphibians, even<br />
for eggs and larvae <strong>of</strong> amphibians with,<br />
again, excellent black and white drawings<br />
<strong>of</strong> amphibian larvae. At <strong>the</strong> end a special<br />
Biota '31-2, 197<br />
chapter is devoted to strategies for protection<br />
<strong>of</strong> both groups. There are also<br />
some very interesting chapters about fossils<br />
and archaeological finds, and probably<br />
<strong>the</strong> most complete list <strong>of</strong> references<br />
ever seen in 39 pages!<br />
The Atlas is a valuable source <strong>of</strong> information<br />
for all those who are interested in<br />
amphibians and reptiles. It is not an ordinary<br />
atlas; it is much more! With much<br />
ecological data presented in various<br />
ways, it is also extremely useful as an<br />
ecological hand<strong>book</strong>. All European herpetologists<br />
should read this <strong>book</strong>, since it<br />
includes so much information and so<br />
many ideas.<br />
I must point out that <strong>the</strong> Atlas is a real<br />
craftsman's tool. I regret, however, that<br />
at least a few brief English summaries<br />
were not included.<br />
Milan Vogrin
198 Biota 3/i-a,<br />
EDITOR ACKNOWLEDGEMENTS<br />
I would like to make <strong>full</strong> acknowledgement <strong>of</strong> <strong>the</strong> generosity to <strong>the</strong> members <strong>of</strong><br />
Editorial Board and <strong>of</strong> <strong>the</strong> referees in both <strong>the</strong>ir time and expertise. I should like to thank<br />
all those who have sent in colour slides for possible reproduction on <strong>the</strong> covers.<br />
List <strong>of</strong> referee (alphabetical order) for volume 1, 2 and 3<br />
1. Maurizio Biondi (Coppito, Italy)<br />
2. Luigi Boitani (Rome, Italy)<br />
3. Massimo Capula (Rome, Italy)<br />
4. Dan Cogalniceanu (Bucharest, Romania)<br />
5. Jelka Crnobrnja-lsailovic (Belgrade, FR Yugoslavia)<br />
6. Helmut Faber (Graz, Austria)<br />
7. Simone Fattorini (Rome, Italy)<br />
8. Paolo Galeotti (Pavia, Italy)<br />
9. Justin Gerlach (Cambridge, United Kingdom)<br />
10. Giinter Gollmann (Wien, Austria)<br />
11. Richard Griffiths (Kent, United Kingdom)<br />
12. Kurt Grossenbacher (Bern, Switzerland)<br />
13. Ulrich Joger (Darmstadt, Germany)<br />
14. Janusz Kloskowski (Lublin, Poland)<br />
15. Zoltan Korsos (Budapest, Hungary)<br />
16. Luca Luiselli (Rome, Italy)<br />
17. Jean Christophe de Massary (Paris, France)<br />
18. Valentin Perez- Mellado (Salamanca, Spain)<br />
19. Juha Merila (Helsinki, Finland)<br />
20. Claude Miaud (Le Bourget du Lac, France)<br />
21. David Mifsud (Basel, Switzerland)<br />
22. Deanna H. Olson (Corvallis, Oregon, USA)<br />
23. Zbynek Rocek (Prague, Czech Republic)<br />
24. Luca Salvati (Rome, Italy)<br />
25. Sabastian Salvido (Genova, Italy)<br />
26. Josef Schmidler (Miinchen, Germany)<br />
27. Andrej Sorgo (Race, Slovenia)<br />
28. Piotr Tryjanowski (Poznan, Poland)<br />
29. Reuven Yosef (Eilat, Israel)<br />
30. Marco Zuffi (Pisa, Italy)<br />
Milan Vogrin
INSTRUCTIONS<br />
TO CONTRIBUTORS<br />
Biota publishes papers from all fields <strong>of</strong><br />
biology and ecology in <strong>the</strong>ir widest<br />
sense.<br />
It is open for authors from all countries.<br />
The language <strong>of</strong> papers is English or<br />
Slovene (with English summary).<br />
Types <strong>of</strong> papers<br />
The journal publishes original scientific<br />
papers, short communications, review<br />
articles, <strong>book</strong> reviews, special issues containing<br />
selected and edited papers dealing<br />
with a specific <strong>the</strong>me or based on a<br />
conference or workshop.<br />
The submission <strong>of</strong> a paper obliges <strong>the</strong><br />
author(s) not to submit <strong>the</strong> same paper<br />
elsewhere. Manuscripts are submitted to<br />
reviewers for evaluation <strong>of</strong> <strong>the</strong>ir significance<br />
and soundness. Authors will generally<br />
be notified <strong>of</strong> acceptance, rejection,<br />
or need for revision within three months.<br />
The referees remain anonymous.<br />
Decisions <strong>of</strong> <strong>the</strong> editor are final.<br />
The form <strong>of</strong> <strong>the</strong> manuscript<br />
The paper should consist <strong>of</strong> title, author's<br />
name and address, abstract (not exceed<br />
250 words), key words (no more than six)<br />
main text (i.e. introduction, material and<br />
methods, results, discussion) followed by<br />
acknowledgements, references, tables<br />
and figures. Authors <strong>of</strong> scientific taxa<br />
should be omitted. Manuscripts should<br />
be double-spaced on one side <strong>of</strong> <strong>the</strong><br />
paper, with wide margins all round. The<br />
text <strong>of</strong> a manuscript should be without<br />
special style settings and footnotes.<br />
Headings should be on separate line, and<br />
<strong>the</strong>ir hierarchy should not exceed three.<br />
Authors are urged to have <strong>the</strong> manuscript<br />
revised by "native speaker", if necessary.<br />
Tables and figures should be numbered<br />
consecutively according to <strong>the</strong> text. Each<br />
should be on a separate sheet <strong>of</strong> paper.<br />
Biota 3/1-2,2002 199<br />
Each caption to a figure or table should<br />
start with <strong>the</strong> number <strong>of</strong> <strong>the</strong> figure/table,<br />
e.g. Figure 1., Table 2. Tables and figures<br />
should be numbered in order <strong>of</strong> appearance<br />
in <strong>the</strong> text.<br />
Illustrations must be submitted cameraready<br />
for copying, usually as laser printouts.<br />
All internal structures, letters,<br />
graphic symbols must be well readable (at<br />
least 2 mm high) after size reduction.<br />
References. In <strong>the</strong> list <strong>of</strong> references, <strong>the</strong><br />
following usage should be followed:<br />
Journal: SURNAME, A.B. Year: Title. Full<br />
Journal name, Volume: pagination.<br />
Book: SURNAME, A.B. & SURNAME, C.<br />
Year: Title. Publisher, Place.<br />
Chapter: SURNAME, A., SURNAME & B.<br />
SURNAME, C. Year: Chapter title. In:<br />
Editors. Book or Proceeding title.<br />
Publisher, Place: pagination.<br />
In <strong>the</strong> text, citations should give <strong>the</strong><br />
author's name and <strong>the</strong> year <strong>of</strong> publication,<br />
e.g. Surname (1998) or (Surname<br />
1998) or Surname & Surname (1998).<br />
Where <strong>the</strong>re are three or more authors,<br />
<strong>the</strong> first authors name plus "et al" must<br />
be given, e.g. Surname et al. (1998,<br />
1999). Where two or more papers abbreviate<br />
to <strong>the</strong> same citation (i.e. two or<br />
more papers produced by <strong>the</strong> same<br />
authors in <strong>the</strong> same year), use "a", "b",<br />
"c", etc. in <strong>the</strong> order <strong>of</strong> <strong>the</strong>ir first appearance,<br />
e.g. (Surname 1998a, b).<br />
Reprints and pro<strong>of</strong><br />
Pro<strong>of</strong> will be send to <strong>the</strong> main author<br />
only once and it should be returned to<br />
<strong>the</strong> Editor without delay. Corrections<br />
should be limited to typographical errors.<br />
The editors reserve <strong>the</strong> right to correct<br />
<strong>the</strong> pro<strong>of</strong>s <strong>the</strong>mselves, using <strong>the</strong> accepted<br />
version <strong>of</strong> <strong>the</strong> typescript, if <strong>the</strong><br />
author's corrections are overdue and <strong>the</strong><br />
journal would o<strong>the</strong>rwise be delayed.<br />
Pro<strong>of</strong>s should be checked very care<strong>full</strong>y. It<br />
is <strong>the</strong> correspondence author's responsi-
NAVODILO AVTORJEM<br />
Biota objavlja prispevke s podrocja<br />
biologije in ekologije v najsirsem pomenu<br />
besede. Prispevki so lahko v angleskem<br />
ali v slovenskem jeziku z daljsim<br />
angleskim povzetkom (enako naslovi<br />
tabel in grafov), ki ga pripravi avtor sam.<br />
Vrste prispevkov<br />
V Bioti je mogoce objavljati izvirne<br />
znanstvene clanke, kratke notice, pregledne<br />
clanke in predstavitve novih knjig.<br />
Biota sprejema tudi prispevke z razlicnih<br />
konferenc in posvetov. Z objavo se avtorji<br />
obvezejo, da ne bodo nikjer objavili<br />
enakega prispevka. Vsi prispevki bodo<br />
predlozeni v recenzijo dvema recenzentoma.<br />
Recenzenti ostanejo anonimni.<br />
Avtorji bodo obvesceni o sprejemu, zavrnitvi<br />
ali reviziji prispevka predvidoma v<br />
treh mesecih. Odlocitev urednika je<br />
dokoncna.<br />
Oblika prispevka<br />
Prispevek mora vsebovati naslov, imena<br />
avtorjev in njihove naslove, izvlecek (do<br />
250 besed), kljucne besede (do sest) ter<br />
glavni tekst (uvod, material in metode,<br />
rezultati, diskusija) ki mu sledi zahvala,<br />
literatura, tabele in grafi. Prispevek predlozite<br />
v dveh izvodih, z dvojnim medvrstnim<br />
razmakom in s sirokimi robovi.<br />
Odstavki naj bodo med seboj loceni s<br />
prazno vrstico. V kolikor je prispevek<br />
pisan v angleskem jeziku, avtorju priporocamo,<br />
da ga pregleda "native<br />
speaker".<br />
Tabele in slike (grafi, fotografije, risbe)<br />
naj bodo ostevilcene po zaporedju, kot se<br />
pojavljajo v besedilu. Vsaka tabela in graf<br />
morata biti na svojem listu. Tabela ali graf<br />
se morata priceti z zaporedno stevilko,<br />
npr. Slika 1, Tabela 2.<br />
llustracije naj bodo tiskane na laserski<br />
tiskalnik. Risbe so lahko narisane tudi s<br />
Biota 3/1-2,2002 201<br />
crnim tusem na paus papirju. Crke,<br />
stevilke in simboli morajo biti velike vsaj 2<br />
mm.<br />
Literatura<br />
Vsi uporabljeni viri morajo biti citirani<br />
med tekstom. Literaturo uredite po<br />
abecednem redu prvega avtorja in glede<br />
na letnico izdaje:<br />
Revija: PRIIMEK, A.B. Leto: Naslov. Polno<br />
ime revije, letnik: strani.<br />
Knjiga: PRIIMEK, A.B. & PRIIMEK, C.<br />
Leto: Naslov. Izdajatelj, kraj.<br />
Poglavje: PRIIMEK, A., PRIIMEK, B. &<br />
PRIIMEK, C. Leto: Naslov poglavja. V:<br />
Urednik(i). Naslov knjige ali zbornika.<br />
Izdajatelj, kraj: strani.<br />
V tekstu citiramo na naslednji nacin:<br />
Priimek (1998) ali (Priimek 1998) ali<br />
Priimek & Priimek (1998). Ce so vec kot<br />
trije avtorji pa: Priimek et al. (1998,<br />
1999). V primeru, ce citiramo vec del<br />
istega avtorja, objavljenih v enem letu,<br />
posamezno delo oznacimo s crkami a, b,<br />
c, itd., npr. (Priimek 1998a, b).<br />
Korektura in separati<br />
Prvi odtis prispevka urednik poslje<br />
glavnemu avtorju v korekturo. Avtor je<br />
dolzan vrniti popravljeno besedilo v<br />
najkrajsem moznem casu. Sirjenje obsega<br />
besedila ob korekturah ni dovoljeno.<br />
Glavni avtor prejme 30 separatov in izvod<br />
revije, kjer je bil objavljen prispevek, brezplacno.<br />
Original in dve kopiji prispevka vkljucno s<br />
tabelami, slikami in grafi posljite na<br />
naslov (v kolikor prispevek posiljate po<br />
elektronski posti, datoteko shranite kot<br />
"obogateno besedilo" - Rich Text Format<br />
(rtf.):<br />
BIOTA<br />
Milan Vogrin, Zg. Hajdina 83c,<br />
SI-2288 Hajdina, Slovenia<br />
Fax: 02 788 30 51<br />
E-mail: milan.vogrin@guest.ames.si
Klub svetnic in svetnikov LDS Zalec<br />
POKRAJINSKI PODJETNISKI FORUM<br />
Ul. Ivanke Uranjek 1, Zalec
Drustvo za proucevanje ptic<br />
in varstvo narave<br />
Society <strong>of</strong> bird research<br />
and nature protection<br />
Ptujska c. 91<br />
SI - 2327 Race<br />
Slovenia
V P<br />
Drustvo varuhov okoija Radoziv<br />
Environmental society Radoziv<br />
Ul. Ivanjke Uranjek 1<br />
SI-3310 Za/ec<br />
Slovenia
forum<br />
Danube Forum<br />
Hanulova 5/D<br />
844 40 Bratislava<br />
www.de-forum.org
TO ADVERTISE IN THIS JOURNAL<br />
SEND AN E-MAIL TO<br />
milan.vogrin@guest.arnes.si