Gyraulus parvus (Mollusca: Gastropoda) in the Czech Republic

Acta Soc. Zool. Bohem. 66: 81–84, 2002ISSN 1211-376XGyraulus parvus (Mollusca: Gastropoda) in the Czech RepublicLuboš BERAN 1) & Michal HORSÁK 2)1)Kokoínsko Protected Landscape Area Administration, eská 149, CZ–276 01 Mlník;e-mail: kokorinsko@proactive.cz, Czech Republic2)Department of Zoology and Ecology, Faculty of Science, Masaryk University, Kotláská 2, CZ–611 37Brno; e-mail: horsak@sci.muni.cz, Czech RepublicReceived April 12, 2001; accepted October 16, 2001Published June 28, 2002Abstract. Next non-native species of aquatic gastropod Gyraulus parvus (Say, 1817) is recorded from theCzech Republic for the first time. Distribution of the species is documented from 12 localities in Bohemia(9) and Moravia (3).Distribution, Mollusca, Gastropoda, Gyraulus parvus, Palaearctic regionINTRODUCTIONGyraulus parvus (Say, 1817) is species native in North America. In Europe this species was foundfirst time in 1973 in Germany (Glöer & Meier-Brook 1998). Recently, only in eastern part of Germanyexist about 15 localities of this species (Glöer & Meier-Brook 1998). The species is very similar tonative Gyraulus laevis (Alder, 1813). They are sibling species according to Meier-Brook (1983).The main diagnostic characters separating G. parvus from G. laevis are as follows (Figs 2, 3):elevated penultimate whorl, marked widening of the distal half of the vas deferens as comparedwith that of the proximal half, distal portion of the spermoviduct is not wider than the widestportion of the sperm duct, and euroekous character of life strategy (Meier-Brook 1983).RESULTSFirst specimens were identified by the senior author in the sample from the sandpit near Lahoš(the sandpit on the right side of the road Lahoš – Teplice, Northern Bohemia, code of mappingsquare 5348[cf. Buchar 1982], November 11, 1999, lgt. L. Beran) and from Barbora (a large waterreservoir or sandpit ? near Teplice, Northern Bohemia, 5348, November 11, 1999, lgt. L. Beran).Material of conchs of “Gyraulus laevis” from similar localities (sandpits) in collections of bothauthors was revised, and after it some localities were visited again and other suitable places wereinvestigated. Other 10 localities with occurrence of Gyraulus parvus resulted from this activity.These localities are as follows – Horka nad Moravou, the sand-pit Podbrady, Central Moravia,6369, April 25, 2000, lgt. L. Beran; Chomoutov, a big sand-pit in the Chomoutovské jezero NatureReserve, Central Moravia, 6369, April 25, 2000, lgt. L. Beran; Ostrá, a sandpit on the right side of theroad Ostrá – Kostomlaty nad Labem, Central Bohemia, 5855, May 7, 2000, lgt. L. Beran; OstrožskáNová Ves, nothern part of the biggest sandpit near Ostrožská Nová Ves, Southern Moravia, 6970,May 7, 2000, lgt. M. Horsák (in Beran & Horsák 1998 determined as G. laevis); Horní Jietín, waterreservoir at former Dolní Jietín village, Northern Bohemia, 5447, October 17, 2000, lgt. L. Beran;Louka u Litvínova, a pond between road and railway at northern border of Louka u Litvínova,Northern Bohemia, 5447, October 18, 2000, lgt. L. Beran; Mariánské Radice, a water reservoir to81

Fig. 3. Gyraulus parvus (Say), mrp – penis retractor muscle, prp – preaputium, psh – penis sheat, dh – distal halfof vas deferens, ph – proximal half of vas deferens, ut – uterus. Scale = 1 mm. Orig. M. Horsák.the south from Mariánské Radice, Northern Bohemia, 5447, October 18, 2000, lgt. L. Beran; MariánskéRadice, a small pond at left side of the road Mariánské Radice – Braany to the east fromMariánské Radice, Northern Bohemia, 5448, October, 18, 2000, lgt. L. Beran; Chomutov, a waterreservoir between Spoice and Droužkovice (nearest to Droužkovice), Northern Bohemia, 5546,October 20, 2000, lgt. L. Beran; Chomutov, small elongated water reservoir between Spoice andDroužkovice (third from Spoice), Northern Bohemia, 5546, October 20, 2000, lgt. L. Beran (see alsothe map on Fig. 1).Very abundant occurrence (at least 100 individuals per square meter) were documented atlocalities Lahoš, Horka nad Moravou and Louka u Litvínova, at other localities were documentedonly scattered (less than 10 specimens per square meter) or solitary (less than 1 specimen persquare meter) occurrence of this species. This North-American species probably inhabits moreespecially artificial (sandpits, water reservoir at reclaimed land) localities on the territory of theCzech Republic, but because of its similarity with Gyraulus laevis it is possible that this species isconfuse with this native species by most of malacozoologists especially in case, when determinationis based only on conchs.83

REFERENCESBERAN L. & HORSÁK M. 1998: Aquatic molluscs (Gastropoda, Bivalvia) of the Dolnomoravský úval lowland,Czech Republic. Acta Soc. Zool. Bohem. 62: 7–23.BUCHAR J. 1982: Publication of faunistic data from Czechoslovakia. Vst. s. Spole. Zool. 46: 317–318.GLÖER P. & MEIER-BROOK C. 1998: Süsswassermollusken (Ein Bestimmungsschlüssel für die BundesrepublikDeutschland) 12. Auflage. Hamburg: Deutscher Jugendbund für Naturbeobachtung, 136 pp.MEIER-BROOK C. 1983: Taxonomic studies on Gyraulus (Gastropoda: Planorbidae). Malacologia 24(1–2): 1–113.84

Acta Soc. Zool. Bohem. 66: 85–97, 2002ISSN 1211-376XOn the morphology and surface ultrastructure of some parasitic nematodes(Nematoda) of birds (Aves)Denisa FRANTOVÁInstitute of Parasitology, Academy of Sciences of the Czech Republic, Branišovská 31,CZ–370 05 eské Budjovice, Czech RepublicDepartment of Parasitology, Faculty of Biological Sciences, University of South Bohemia, Branišovská 31,CZ–370 05 eské Budjovice, Czech RepublicReceived February 1, 2001; accepted October 16, 2001Published June 28, 2002Abstract. The morphology of four species of avian nematodes was studied using scanning electronmicroscopy (SEM): Porrocaecum depressum (Zeder, 1800) from Buteo buteo; third- and fourth-stagelarvae of Porrocaecum semiteres (Zeder, 1800) from Larus ridibundus and Turdus philomelos; Acuariaanthuris (Rudolphi, 1819) from Corvus frugilegus; Cosmocephalus obvelatus (Creplin, 1825) from Larusridibundus. The examination of the head end of adult Porrocaecum depressum and fourth-stage larvae ofP. semiteres revealed a pattern of labial papillae that is typical of ascaridoid genera. The structure of thehead end of third- and fourth-stage larvae of P. semiteres seems to be identical with that of the related P.ensicaudatum, which occurs in the same species of intermediate and definite host. The fourth-stage larvaeof P. semiteres was redescribed. A detailed examination of the oral region of Acuaria anthuris revealed teethat the anterior end of the cordons, which may serve to host tissue damage during feeding. The deirids of A.anthuris are very small, with a bicuspid tip.Morphology, surface ultrastructure, nematode, parasite, AvesINTRODUCTIONThe taxonomy of parasitic nematodes of birds (Aves) is often based on incomplete descriptionsand drawings. During a recent study of some materials from birds (Frantová 2002) provided by theHelminthological Collection of the Institute of Parasitology, Academy of Sciences of the CzechRepublic (ASCR), in eské Budjovice, morphology of four common species of nematodes wasstudied in detail, using light microscopy and scanning electron microscopy (SEM): Porrocaecumdepressum, P. semiteres, Acuaria anthuris and Cosmocephalus obvelatus.Porrocaecum depressum is a common parasite of birds of prey (Falconiformes, Strigiformes)and its morphology had been described several times using light microscopy (see Mozgovoy1953, Hartwich 1975). P. semiteres is typical of birds of the order Charadriiformes. It frequentlyoccurs as a third- or fourth-stage larva in the digestive tract of small passerines (especially Turdidae),but is unable to mature in these atypical hosts (Iygis 1967). Descriptions and drawings ofthird-stage larvae are included in the works of Mozgovoy & Bishaeva (1959), Iygis (1967) andMoravec (1971). There are few accounts of the morphology of the fourth-stage larvae (Iygis 1967).A related species, P. ensicaudatum (Zeder, 1800), also uses small passerines (especially Turdidae)as definitive hosts, but is also found as larvae in Charadriiformes (Iygis 1967, 1970). Consequently,larvae of P. ensicaudatum and P. semiteres can occur in the same species of avian host. They canbe distinguished by the ratio of the stomach to the intestinal caecum length (Supryaga & Supryaga1971, Baruš et al. 1978b). In the genus Porrocaecum Railliet et Henry, 1912, only P. ensicaudatumwas examined using SEM (Wharton 1978, Baruš et al. 1983, McNeill & Anderson 1990a, b).85

Tab. 1. Nematodes studied with the aid of SEMnematode species host site localityPorrocaecum depressum Accipiter gentilis duodenum eské Budjovice (1980)Buteo buteoPorrocaecum semiteres, Larus ridibundus gizzard Praha (1978)3 rd - and 4th-stage larvae Turdus philomelos Klec (1982, 1983)eské Budjovice (1999)Acuaria anthuris Corvus frugilegus L. gizzard eské Budjovice (1981, 1982)Cosmocephalus obvelatus Larus ridibundus oesophagus KlecThe ultrastructure of the head end larval P. ensicaudatum was studied by McNeill & Anderson(1990a, b). The present study focuses on the head end of larval P. semiteres with the aim of findingdifferences.Acuaria anthuris is a frequent parasite in gizzards of corvids (Corvidae). There are several lightmicroscopy studies that deal with the morphology of this species (see Chabaud & Petter 1961,Skryabin et al. 1965, Baruš et al. 1972). The cordon ultrastructure was studied by Baruš & Majumdar(1975).Cosmocephalus obvelatus is a parasite of the oesophagus of various piscivorous birds. Itsmorphology has been studied using light microscopy (Skryabin et al. 1965, Baruš et al. 1978a,Anderson & Wong 1981). SEM micrographs of second- and third-stage larvae of C. obvelatuswere published by Wong & Anderson (1982).MATERIALS AND METHODSAll of the nematodes studied were provided by the Helminthological Collection of the Institute of Parasitology,ASCR, eské Budjovice. The parasites were collected from birds shot at localities in South Bohemia, CzechRepublic, from1977–1983 (Tab. 1). They were fixed and stored in 4% formaldehyde and, after being cleared inglycerine, were identified under an optical microscope. Drawings were made with the aid of a Zeiss microscopedrawing attachment. Prior to SEM examination, the specimens selected were washed in 4% formaldehyde,postfixed for 2 hours in 2% aqueous osmium tetraoxide, washed in double-distilled water, dehydrated through aseries of increasing concentrations (10–100%) of ethyl alcohol, critical point dried using CO2 and coated withgold palladium in the POLARON PS 100 sputter coater. The JEOL SEM-6300 scanning electron microscope wasused to examine and photograph the specimens.RESULTSFamily Ascarididae Baird, 1853Porrocaecum depressum (Zeder, 1800) Baylis, 1920(Fig. 1)DESCRIPTION (based on SEM examination of the head ends of 2 males and 2 females). Oral openingtriradiate, surrounded by three massive labia (one dorsal and two subventral) and three smallFig. 1. Porrocaecum depressum (Zeder) from Buteo buteo. SEM micrographs. A – head end, apical view; B – innermargin of labium with detail of teeth; C – subventral labium, ventrolateral view. Note double papilla (dp), singlepapilla (sp) and amphid (a); D – dorsal labium, dorsal view. Note double papillae (dp).Fig. 2. Porrocaecum semiteres (Zeder) from Larus ridibundus (A) and P. ensicaudatum (Zeder) from Turdusphilomelos (B). Third-stage larvae, lateral view. Scale bar in mm.86

Fig 3. Porrocaecum semiteres (Zeder) from Turdus philomelos (A, B) and Larus ridibundus (C–F). SEM micrographs.A – cephalic extremity of third-stage larva, apical view. Note subdorsal and subventral papillae (arrows),amphids (a) and ventral tooth (vt); B – cephalic extremity of 3 rd stage larva, ventral view. Note ventral tooth (vt);C – head end of 4 th stage larva, apical view; D – lateral alae of 4 th stage larva; E – dorsal labium of 4 th stage larva.Note two double papillae (dp); F – subventral labium of 4 th stage larva. Note double papilla (dp), single papilla (sp)and amphid (arrow).88

Tab. 2. The dimensions (mm) of third- and fourth-stage larvae of Porrocaecum semiteres (Zeder, 1800) fromLarus ridibundus and Turdus philomelosthird-stage larvae fourth-stage larvaemalefemalebody length 3.448 (3.376–3.520) 8.08 (6.82–9.34) 7.952 (3.712–12.192)maximum width 0.088 (0.080–0.096) 0.216 (0.144–0.288) 0.200 (0.112–0.288)nerve ring – anterior 0.217 (0.213–0.221) 0.300 (0.271–0.328) 0.289 (0.210–0.368)excretory pore – anterior 0.219 (0.218–0.220) 0.364 (0.328–0.400) 0.351 (0.253–0.448)length of muscular oesophagus 0.419 (0.410–0.428) 1.016 (0.784–1.248) 0.862 (0.435–1.288)length of ventriculus 0.054 (0.045–0.063) 0.128 (0.105–0.151) 0.113 (0.076– 0.150)intestinal caecum 0.038 (0.030–0.045) 0.059 (0.050–0.068) 0.108 (0.068–0.148)ratio of length of ventriculusto that of caecum 1:0.70 (1:0.67–0.71) 1:0.67 (1:0.48–0.85) 1:0.9 (1:0.63–1:1.23)genital primordium – anterior 1.746 (1.723–1.768)vulva – anterior 4.188 (2.096–6.280)length of spicule primordia 0.134 (0.068–0.200)tail 0.145 (0.126–0.164) 0.164 (0.126–0.202) 0.199 (0.125–0.272)interlabia (Fig. 1A). Cuticle of head end smooth. Labia hexagonal in shape, very narrow at base,widest in middle. Anterior half of labia divided by longitudinal shallow median depression into twolobes. Each lobe supported by a finger-like projection, bluntly terminating close to anterior marginof labium. Anterior margins of labia rounded, rimmed with single row of teeth (Fig. 1B). Teethconical, with rounded tips, usually longer than wide, expanding laterally and caudally to last thirdof labia. Dorsal labium with two double papillae situated mediolaterally (Fig. 1D). Each subventrallabium with one double papilla situated medioventrally, and a single papilla and amphid situatedanterolaterally (Fig. 1C). Interlabia indistinct, triangular in shape, reaching first third of labia.COMMENT. Based on light microscopy, the specimens studied showed the dimensions and morphologyconsistent with the description given by Mozgovoy (1953)Porrocaecum semiteres (Zeder, 1800) Baylis, 1920(Figs 2, 3, 4; Tab. 2)DESCRIPTION. Third-stage larvae: 10 specimens studied using light microscopy (Fig. 2, Tab. 2), 2 ofwhich were used for SEM (Figs 3A, B). Anterior end bluntly rounded, with smooth roundedcircular rim of cuticle (Fig. 3A). Labia absent. Two single cephalic papillae subdorsally and twosubventrally at base of cuticular rim. Two amphids situated laterally on either side. One ventraltooth present (Fig. 3A, B). Oral opening circular. Cuticle with distinct annulation and lateral alae.Excretory porus slightly posterior to nerve ring. Oesophagus with anterior muscular portion andposterior ventriculus. Intestine dark, with rudimentary, anteriorly directed dorsal caecum. Caecumlonger than half the length of ventriculus. Ventral genital primordium positioned slightly posteriorto mid body. Sexes not distinguishable. Anus subventral. Tail tapered and pointed.COMMENT. Based on light microscopy, the specimens showed the dimensions and morphologyconsistent with the description given by Iygis (1967) and Okulewicz (1979). Larvae of P. semitereswere distinguished from those of the related P. ensicaudatum on the basis of the ratio of ventriculusand intestinal caecum lengths by Supryaga & Supryaga (1971) and Baruš et al. (1978b) (Fig.2, Tab. 2).89

Fig. 6. Acuaria anthuris (Rudolphi) from Corvus frugilegus. SEM micrographs. A – tail of female, ventral view.Note phasmids (arrows); B – caudal extremity of male, ventral view. Single median precloacal papilla and threepairs of postcloacal papillae; C – tail of male, ventral view. Two pairs of postcloacal papillae and single pair ofterminal phasmids; D – cloacal region of male. Note distal end of spicule (sp) and single median precloacal papilla(arrow).92

Family Acuariidae Railliet, Henry et Sisoff, 1912Acuaria anthuris (Rudolphi, 1819) Railliet, Henry et Sisoff, 1912(Figs 5, 6)DESCRIPTION (based on SEM examination of 2 male and 2 female adult specimens). Medium sizednematodes. Males smaller than females, with ventrally coiled tail. Cuticle with prominent annulation.Head end with two lateral pseudolabia, separated from rest of body by a transverse groove(Figs 5B, D, E). Pseudolabia massive, with smooth cuticle, triangular in shape, widest at base,tapering anteriorly to rounded tip. Each pseudolabium medially divided by a longitudinal ridge,extending approximately to second third of length of pseudolabia, to level of shallow transversegroove separating tips. Single prominent papillae laterally on both sides of pseudolabium (Figs 5C,E). Amphid situated medially in middle of pseudolabium (Fig. 5C). Tips free, apparently mobile.Distinct curve formed dorsally and ventrally at inner edge of each tip, giving it a nose-like appearance(Fig. 5E). Cordons in form of two laterodorsal and two lateroventral longitudinal cords,slightly exceeding body surface and separated from neighbouring cuticle by a deep grooves. Eachcordon consisting of two parallel unconnected cords, transverselly divided into prominent ridgescorresponding with annulation of neighbouring cuticle; first ridge at anterior end of both cords inform of tooth (Fig. 5E). Cordons beginning dorsally (ventrally) at level of curved inner edge ofpseudolabial tip, then running between psedolabia and diverging laterodorsally (lateroventrally)at their bases; extending to second third of body. Deirids small, poorly visible, with bicuspid tip(Fig. 5F). Phasmids terminal (Figs 6A, C). Caudal extremity curved ventrally (Fig. 6B). Caudal lateralalae present, bearing four pairs of preanal and six pairs of postanal papillae. Two pairs of postanalpapillae just posterior to cloaca, a solitary pair in middle of tail and three pairs near tip of tail (Figs6B, C). Single papilla anteriomedian to cloaca (Fig. 6D).COMMENT. The specimens were larger than with the dimensions given by Baruš et al. (1972) fornematodes from the same host.Cosmocephalus obvelatus (Creplin, 1825) Seurat, 1919(Figs 7, 8)DESCRIPTION (based on SEM examination of 2 males and 1 female specimen). Medium sized nematodes.Cuticle with fine annulation. Lateral alae present, beginning at level of deirids and extendingto posterior quarter of body (Fig. 7A). Two rather small lateral pseudolabia, rhomboid in shape,widest in middle, narrowing towards tips and bases (Figs 7B, C, G). Tips rounded and protruding(Figs 7D, E); two median ridges separating dorsal and ventral pore on each tip (Fig. 7F). Medianpart of pseudolabium convex, bearing prominent amphid (Fig. 7G). Median margins of pseudolabiaprojecting dorsally and ventrally into lobes (Fig. 7C), each with indistinct papilla. Two conspicuouscordons (dorsal and ventral) restricted to head end, with dense transverse striation; beginningdorsally (ventrally) to mouth, then copying shape of pseudolabia, running posteriorly, recurrentand anastomosing (Fig. 7B). Deirids large, with bicuspid tip (Fig. 8C). Excretory porus situatedventrally, slightly posterior to level of deirids (Fig. 8A). Prominent phasmids, situated subventrallynear tip of tail (Fig. 8B). Caudal extremity of male with caudal alae bearing four pairs of preanal andfive pairs of postanal pedunculate papillae (Figs 8D, E). Two pairs of small sessile papillae presentat tip of tail, near last pair of pedunculate papillae. Two spicules, dissimilar in size and shape: rightspicule short and massive, left long and slender. Caudal extremity of female tapered and round (Fig.8B). Knob-like projection at tip of tail, clearly visible in light microscopy, indistinct (probablyshrank or damaged during preparation).93

COMMENT. These specimens were studied previously (Frantová 2002); they are smaller comparedthan the dimensions given by Anderson & Wong (1981).DISCUSSIONAscarididaeThe present SEM study of the head end ultrastructure of Porrocaecum depressum confirmed, withfew exceptions, the descriptions given by Mozgovoy (1953) and Hartwich (1975). There is adifference in the description of labial papillae, which are small and indistinct in this species,compared, for example, with P. ensicaudatum (Baruš et al. 1983, McNeill & Anderson 1990a, b).The papillae on the dorsal labium and the medioventral papilla on each subventral labium aredouble. There are a single papilla and an amphid anterolaterally on both subventral labia. Thepattern of papillae is consistent with the above mentioned light microscopy studies and similar inall ascaridoid genera.The head end ultrastructure of third- and fourth-stage larvae of P. semiteres did not differ fromthat of P. ensicaudatum (McNeill & Anderson 1990a, b). The cuticular rim with four single papillae,two ampids and a single ventral tooth in the third-stage larvae is replaced by three labia with fourdouble and two single papillae, and two amphids in the fourth-stage larvae. The fourth-stagelarvae have a pattern of labial papillae similar to that of the adults. The head end structure of adultP. semiteres, as seen in light microscope, seems to be almost identical with that of P. ensicaudatum(see Mozgovoy 1953, Hartwich 1975). Adults of both genera can be distinguished as lateral alaeare absent in adult P. ensicaudatum and the ratio of the length of the ventriculus to that of theintestinal caecum is 1: 0.64–1.2 in P. semiteres (Baruš et al. 1978b) and 1 : 0.15–0.36 in P. ensicaudatum(Supryaga & Supryaga 1971). The ratio of the length of the ventriculus to that of the intestinalcaecum seems to be the only difference between larval P. ensicaudatum and P. semiteres.AcuariidaeAcuaria anthuris has been described several times using ligth microscopy (Chabaud & Petter1961, Skryabin et al. 1965, Baruš et al. 1972). These descriptions differ mainly in the number ofpostanal papillae in the male. Chabaud & Petter (1961) stated that the number of the latter varieddepending on the host: those from Pica and Garrulus had 6 pairs of postcloacal papillae and onepair of terminal phasmids; those from Corvus mostly 7 pairs of postcloacal papillae and one pair ofphasmids, with rare specimens that have the same pattern as in Pica and Garrulus. Baruš et al.(1972) observed 7–8 pairs of postcloacal papillae in the nematodes parasitic in Corvus frugilegus.The present SEM study of specimens from C. frugilegus, revealed 6 pairs of postcloacal papillaeand one pair of terminal phasmids in males; there were no difference in the specimens examinedusing light microscopy. The ultrastructure of the cordons was studied by Baruš & Majumdar(1975). The present study completes the previous description by adding details of the organizationof the cordons at the level of the oral opening. The anterior ends of cordons project into teeth,which may serve to host tissue damage during feeding. Data on deirids is scarce, only theirposition has previously been mentioned (see Skryabin et al. 1965). The bicuspid tips are typical ofC. obvelatus and other acuariid species, but there can be intraspecific variations in the shape ofdeirids and the number of tips (Moravec, unpubl. data). The SEM study of C. obvelatus fullyconfirmed the data presented by Anderson & Wong 1981).94

Fig. 7. Cosmocephalus obvelatus (Creplin) from Larus ridibundus. SEM micrographs. A – cephalic extremity,dorsoventral view. Note deirids (arrows) and lateral ala (la); B – head end, lateral view; C – detail of pseudolabium,lateral view; D – detail of pseudolabia, dorsoventral view; E – head end, dorsoventral view; F – detail of pseudolabia,apical view. Note pores (arrowhead) and amphid (a); G – head end, apical view.95

Fig. 8. Cosmocephalus obvelatus (Creplin) from Larus ridibundus. SEM micrographs. A – excretory pore, ventralview; B – caudal extremity of female, subventral view. Note phasmid (arrow); C – deirid, lateral view; D – caudalextremity of male, ventral view; E – caudal extremity of male, sublateral view. Note distal end of left (slender)spicule.96

A c k n o w l e d g e m e n t sThe author thanks F. Moravec of the Institute of Parasitology, ASCR, in eské Budjovice for providing thenematodes from the Helminthological collection and critical comments, and the workers of the Laboratory ofElectron Microscopy, ASCR, in eské Budjovice for preparing the nematodes for SEM examination.REFERENCESANDERSON R. C. & WONG P. L. 1981: Redescription of Cosmocephalus obvelatus (Creplin, 1825) (Nematoda:Acuarioidea) from Larus delawarensis Ord (Laridae). Can. J. Zool. 59: 1897–1902.BARUŠ V. & MAJUMDAR G. 1975: Scanning electron microscopic studies on the cordon structures of seven acuariidgenera (Nematoda: Acuariidae). Folia Parasitol. 22: 125–131.BARUŠ V., RYŠAVÝ B., GROSCHAFT J. & FOLK . 1972: The helminth fauna of Corvus frugilegus L. (Aves, Passeriformes)in Czechoslovakia and its ecological analysis. Acta Sci. Natur. Brno 6: 1–53.BARUŠ V., SERGEEVA T. P., SONIN M. D. & RZYHIKOV K. M. 1978a: Helminths of Fish-Eating Birds of thePalaearctic Region I. Nematoda. Praha: Academia, 318 pp.BARUŠ V., SITKO J. & TENORA F.1978b: Nematoda and Pentastomida parasiting gulls (Aves: Laridae) in Bohemiaand Moravia. Acta Sci. Natur. Brno 26: 169–189.BARUŠ V., TENORA F., RYŠAVÝ B. & WIGER R. 1983: Morphology and ultrastructure of the head end of Porrocaecumensicaudatum. Vst. s. Spole. Zool. 47: 1–5.CHABAUD A. G. & PETTER A. 1961: Nematodes du genre Acuaria de la faune de France. Ann. Parasit. 36: 409–424.FRANTOVÁ D. (2002): Some parasitic nematodes (Nematoda) of birds (Aves) in the Czech Republic. Acta Soc.Zool. Bohem. 66: 13–28.HARTWICH G. 1975: Die Tierwelt Deutschlands. 62. Teil. I. Rhabditida und Ascaridida. Jena: VEB G. Fisher Verlag,256 pp.IYGIS V. A. 1967: [Life cycle of Porrocaecum semiteres (Zeder, 1800) (Nematoda: Ascaridata)]. Parazitologiya 1:213–218 (in Russian).IYGIS V. A. 1970: [Experimental studies of the host specifity of Porrocaecum ensicaudatum (Zeder, 1800)].Parazitologiya 4: 563–568 (in Russian).MCNEILL M. A. & ANDERSON R. C. 1990a: Development of Porrocaecum ensicaudatum (Nematoda: Ascaridoidea)in terrestrial oligochaetes. Can. J. Zool. 68: 1476–1483.MCNEILL M. A. & ANDERSON R. C. 1990b: Development of Porrocaecum ensicaudatum (Nematoda: Ascaridoidea)in starlings (Sturnus vulgaris). Can. J. Zool. 68: 1484–1493.MORAVEC F. 1971: A new natural intermediate host of the nematode Porrocaecum semiteres (Zeder, 1800). FoliaParasitol. 18: 26.MOZGOVOI A. A. 1953: Askaridaty životnych i eloveka i vyzyvaemye imi zabolevanija. Kniga 2. Osnovynematodologii II. [Ascaridata of Animals and Man and Diseases Caused by Them. Part 2. Principles ofNematology II. ] Moscow: Izdat. AN SSSR, 616 pp (in Russian).MOZGOVOY A. A. & BISHAEVA L. 1959: [On the life cycle of Porrocaecum heteroura (Ascaridata, Anisakidae)].Helminthologia 1: 195–197 (in Russian).OKULEWICZ A. 1979: Threadworms of blackbird (Turdus merula L.) and mavis (Turdus philomelos Brehm) fromthe region of Wrocaw. I. Faunistic study. Wiad. Parazytol. 25: 301–331.SKRYABIN K. I., SOBOLEV A. A. & IVASHKIN V. M. 1965: Spiruraty Životnych i eloveka i Vyzyvaemye ImiZabolevanija. Kniga 3. Acuarioidea. Osnovy Nematodologii XIV [Spirurata of Animals and Man and DiseasesCaused by Them. Part 3. Acuarioidea. Principles of Nematodology XIV] Moscow: Izdat. AN SSSR, 570 pp (inRussian).SUPRYAGA V. G. & SUPRYAGA A. M. 1971: [To the differential diagnosis of the larvae of the nematode genusPorrocaecum Railliet et Henry, 1912 from the intermediate hosts – Oligochaeta.] Tr. GELAN 22: 200–203 (inRussian).WHARTON D. A. 1979: The structure of the egg shell of Porrocaecum ensicaudatum (Nematoda: Ascaridida). Int.J. Parasitol. 9: 127–131.WONG P. L. & ANDERSON R. C. 1982: The transmission and development of Cosmocephalus obvelatus (Nematoda:Acuarioidea) of gulls (Laridae). Can. J. Zool. 60: 1426–1440.97

Acta Soc. Zool. Bohem. 66: 98, 2002ISSN 1211-376XBOOK REVIEWERZINÇLIOGLU Z.: Maggots, Murder and Men (Memories and Reflections of a Forensic Entomologist).Harley Books, Colchester, Essex, UK, 2000, 256 pp.; format 232×150mm. Price 13.95; ISBN 0-946589-65-8Books written by specialists in a field are an invaluable resource. Based on the author’s personal experience, theygive a better perspective than books written as reviews, often by somebody from another field. Maggots, Murder andMen belongs to those that are especially remarkableDr. Zakaria Erzinçlioglu, commonly known in English circles of criminology as “Dr. Zak” or “maggotologue”,has been working in the field of Forensic Entomology for over ten years. He collects and identifies insects foundassociated with human corpses, most notably Dipteran larvae (maggots). His work helps investigators to determinetime of death and whether bodies have been manipulated or moved from the original scene of the crime. Vieweddispassionately, a dead human body is a magnificent and highly nutritious resource for insects as well as otherinvertebrate animals. The book relates several accounts of criminal cases in which the author participated,including the description of sometimes startling circumstances of cases, attempts to influence his reports, the useof “muddy-water” consultants by defence lawyers, e.g. unqualified “experts” without any knowledge of the entomologyinvolved etc. There is also much about the use of entomology in archaeology, the human history of myiasisand other interesting topics.From the book, it is clear there is empathy between the author and the unfortunate victims. Erzinçlioglu’s casesare far from the luxurious environment of criminal TV with detective officer Columbo or attorney Perry Mason,but rather, for example, in a dirty dwelling in a deprived area of London, where a victim’s bodies was secreted underfloorboards. Another feature that is palpable in the book is that the author is a learned and lettered man. Eachchapter begins with a citation and citations are also in the text (Sherlock Holmes!). Chapters are introduced withfine scrapeboard illustrations resembling woodcuts and the book is closed by “sources and further reading” and awell-done index. The book is clearly oriented to a popular audience, but written on the highest professional level,so clearly recommendable to every entomologist or zoologist interested in forensic practice. If it were not on sucha disturbing subject matter, it would be recommendable for recreational reading before sleeping.Josef Chalupský98

Acta Soc. Zool. Bohem. 66: 99–119, 2002ISSN 1211-376XHarpalus larvae (Coleoptera: Carabidae: Harpalina): description of severalspecies and taxonomic remarksKarel HRKA & Zdenk PAPOUŠEKDepartment of Zoology, Charles University, Vininá 7, CZ–128 44 Praha 2, Czech Republic;e-mail: hurka@natur.cuni.cz, brkous@seznam.czReceived June 15, 2001; accepted October 16, 2001Published June 28, 2002Abstract. Three larval instars of Harpalus (Harpalus) atratus Latreille, 1804, H. (H.) cisteloides hurkaiDivoký, Pulpán et Rébl, 1990, H. (H.) picipennis (Duftschmid, 1812), H. (H.) saxicola Dejean, 1829, H.(H.) servus (Duftschmid, 1812), first and third larval instars of H. (H.) luteicornis (Duftschmid, 1812) andthird larval instar of H. (H.) solitaris Dejean, 1829 and H. (H.) xanthopus winkleri Schauberger, 1923 aredescribed and illustrated. Differential diagnosis of the nominotypical subgenus Harpalus Latreille, 1802,based on larval characters, is given. Taxonomic significance of species aggregates and subgenera arediscussed. Some new larval characters (detailed additional chaetotaxy of femora) are used.Larval taxonomy, preimaginal characters, Coleoptera, Carabidae, HarpalusINTRODUCTIONThe subgenus Harpalus Latreille, 1802 consists of about 300 species world-wide (Lorenz 1998).The more or less superficial and incomplete notes on about 30 species of larvae of this subgenusare mentioned in the literature (Schiodte 1867, Gardner 1938, van Emden 1942, Chu 1945, Larsson1941, 1968, Sharova 1958, 1964, Habu & Sadanaga 1963, 1965, 1970a, b, Kirk 1972, Habu 1973, Hrka1975, Brandmayr, Ferrero & Zeto Brandmayr 1980, Arndt 1991, Luff 1993). Only the works by Habu& Sadanaga (1963, 1965, 1970a, b), Habu (1973) and Arndt (1991) deal with more than one larvalinstar and consider the chaetotaxy.The purposes of the paper are to describe to date undescribed larvae of Harpalus, discusspossible relationships between the taxa studied, and provide the differential diagnosis of thesubgenus Harpalus in the larval stage.MATERIAL AND METHODSThree larval instars of Harpalus (Harpalus) atratus (5 L1, 2 L2, 6 L3), H. (H.) cisteloides hurkai (5 L1, 5 L2, 6 L3),H. (H.) luteicornis (5 L1, 5 L3 ), H.(H.) picipennis (6 L1, 4 L2, 6 L3), H. (H.) saxicola (2 L1, 1 L2, 2 L3), H. (H.)servus (18 L1, 3 L2, 6 L3) and the third instar of H. (H.) solitaris (1 L3) and H. (H.) xanthopus winkleri (1 exuviaof L1, 1 L3) are reared ex ovo during years 1989 and 1999 following the technique described by Hrka (1996).The parental pairs are found as follows: H. (H.) atratus – Bohemia centr., Praha-Kunratice (code of mappingsquare 5952, for details see Pruner & Míka 1996), 24.4.1998, Z. Papoušek leg.; H. (H.) cisteloides hurkai –Bohemia, eské stedohoí, Písený vrch, (5548), 11.9.1993, P.Veselý leg.; H. (H.) luteicornis – Bohemia centr.,Praha-Radotín, Cikánka env. (6051), fallow, 20.4.1999, K. Hrka leg.; H. (H.) picipennis – Bohemia centr.,Tuha nr. Neratovice (5753), 21.6.1995, K. Hrka leg.; H. (H.) saxicola – Slovakia mer., Avaš hill nr. Sírnik(7497), 8.4.1989, J. Hejkal leg.; H. (H.) servus – Moravia, Bzenec-stelnice (7069), 28.9. 1997, Z. Papoušek leg.;H. (H.) solitaris Bohemia, eský les, erchov 1000m, (6642), 29.7.1993, K. Hrka leg.; H.(H.) xanthopuswinkleri – Bohemia centr., Travice (5451), 15.5.1997, K. Hrka leg.99

For comparative purpose larvae of following taxa have been studied: H.(Acardystus) flavescens (Piller etMitterpacher, 1783), H. (Harpalus) anxius (Duftschmid, 1812), H. (H.) autumnalis (Duftschmid, 1812), H. (H.)distinguendus (Duftschmid, 1812), H. (H.) froelichii Sturm, 1818, H. (H.) honestus (Duftschmid, 1812), H.(H.)latus (Linnaeus, 1758), H. (H.) pumilus Sturm, 1818, H. (H.) subcylindricus Dejean, 1829, H. (H.) tardus(Panzer, 1797) and H. (Semiophonus) signaticornis (Duftschmid, 1812).All material is deposited in the Collectio Hrka of the Charles University Praha, Department of Zoology. Thenotation of setae and pores follows the papers by Bousquet & Goulet (1984) and Bousquet (1985).DESCRIPTION OF LARVAEHarpalus (Harpalus) atratus Latreille, 1804(Figs 1–10)First instarHABITUS AND COLOR. Head capsule and pronotum conspicuously broad and transverse; bodyyellow, head and pronotum pale brown, retinaculum and apex of mandible dark brown. Width ofhead capsule 1.44–1.50 mm (x = 1.47 mm, n = 5).MICROSCULPTURE. Isodiametric microsculpture absent, granulated microsculpture on pronotal prescutumand postscutum.CHAETOTAXY. Only one additional pair of spines on anterior femora; seta PA4 very fine, but distinctlylonger than setae PA1 – PA3; setae FR1 and FR3 obviously reduced; group gMX consists of 5–6 thicker and more than 70 thinner, long setae.HEAD (Fig. 1) distinctly transverse (index width/length 1.35); nasale (Fig. 2) not or slightly prominent,distinctly toothed throughout, with 13–16 teeth, two lateral on each side larger, ventral rowwith 30–32 teeth; adnasalia moderately sloping; cervical grooves clearly bent, almost reaching thelevel of seta PA7 on dorsal side, don’t reaching the ventral side; coronal suture very long, distinctlylonger than length of antennomere II; egg burster consists of 5–6 small teeth on each side offrontale. Antennae as long as mandibles; mandible (Fig. 3) triangular, 3 times as long as basalwidth, with one larger and one smaller teeth in front of retinaculum; seta MB1 short, only a littlelonger than retinaculum, penicillus present; maxillae (Fig. 4) slender, stipes more than 3 times aslong as wide; labium (Fig. 5) very short and rounded on lateral borders, ligula reaching the level ofone quarter of palpomere I, distinctly bifid at apex.THORAX. Pronotum transverse, as wide as head, notal carina indistinct, seta PR1 reduced up topore; legs (Fig. 6) normally developed, claws unequal, anterior curved, almost twice as long asposterior on the first pair and only a little longer than posterior on the other pairs.ABDOMEN. Urogomphi subparallel, longer than width of tergum IX and of anal tube.Second and third instarsSame character states as in the first instar except for following:HABITUS. Width of head capsule 1.89 and 2.07 mm in the second and 2.45–2.59 mm (x = 2.52 mm,n = 6) in the third instar.MICROSCULPTURE. Isodiametric microsculpture on head and pronotum, granulated microsculptureon pronotal prescutum and postscutum and on postscutum of meso- and metanotum.CHAETOTAXY. Head: 2 secondary setae on inner side of antennomere II, 2 setae on outer side ofstipes, 1–2 setae near apex of palpomere I, 8 pairs of setae (10 pairs in L3) on dorso-lateral surfaceof prementum, 1–2 setae on place of inner and outer stemmatal furrows.Thorax: pronotum with 2–3 secondary setae along notal carina, several setae on lateral margin and between setae PR11 andPR12; 9–11 secondary spines on each side of anterior femora. Abdomen: abdominal tergites with 1secondary seta between TE1 and TE6, 2 setae between TE6 and TE7 and 1–2 setae between TE9100

Figs 1–10. Harpalus (Harpalus) atratus Latreille. First instar larva: 1 – head capsule (dorsal view). 2 – nasale. 3 –mandible. 4 – maxilla. 5 – labium. 6 – first leg. Third instar larva: 7 – nasale. 8 – antenna. 9 – mandible. 10 –urogomphi. Scales: 1 mm (Figs 1, 10), 0.5 mm (Figs 3, 4, 5, 6, 8, 9), 0.2 mm (Figs 2, 7).101

and TE10, 2–3 small setae on lateral margin; 4 long and many small secondary setae on surface ofurogomphi, length of URβ 0.65 times that of UR4, URγ about as long as UR4.HEAD AND NASALE (Fig. 7) similar as in L1, nasale more prominent, lateral teeth larger; cervicalgrooves reaching the level of PA5; coronal suture longer than antennomere II; seta PA4 as long asantennomere II or III; setae PA1,2,3very small; antenna (Fig. 8); mandible with only one extra tooth(Fig. 9).THORAX. Claws unequal as in first instar.ABDOMEN. Urogomphi (Fig. 10) long and subparallel, twice as long as width of tergum IX, distinctlylonger than anal tube.Harpalus (Harpalus) cisteloides hurkai Divoký, Pulpán et Rébl, 1990(Figs 11–19)First instarHABITUS AND COLOR. Body yellow white, head and pronotum pale brown, retinaculum and apex ofmandible dark brown. Width of head capsule 1.14–1.20 mm (x = 1.16 mm, n = 5).MICROSCULPTURE. Isodiametric microsculpture on frontale and parietale, granulated microsculptureon prescutum and postscutum of pronotum and on postscutum of mesonotum, pointed microsculptureon metanotum and on abdominal terga.CHAETOTAXY. Excepting ancestral chaetotaxy one additional seta on dorso-lateral surface of prementumand 4 pairs of additional spines on anterior femora; seta PA4 slightly longer than coronalsuture, as long as antennomere IV and distinctly longer than setae PA1 – PA3; the majority of setaeon frontale, except FR2, relatively short, especially setae FR8,9 and one additional neighbouringseta; group gMX consists of 4–5 thicker and more than 50 thinner, long setae.HEAD (Fig. 11) distinctly transverse (index width/length 1.3); nasale (Fig. 12) protruding, withtriangular central incision and with 5–6 teeth on each side, two lateral only a little larger, ventral rowconsists of 18–22 teeth, adnasalia moderately sloping; cervical grooves clearly bent, almost reachingthe level between setae PA7 and PA4 on dorsal side, don’t reaching the level of seta PA16 onventral side; coronal suture distinctly longer than width of antennomere I; egg burster consists of2 smaller teeth on the level of seta PA4 on each side. Antennae slightly longer than mandibles;mandible (Fig. 13) strongly curved and robust, twice as long as basal width, with only a slightswelling in front of retinaculum, penicillus present; maxillae (Fig. 14) slender, stipes almost 3.5times as long as wide; labium (Fig. 15) rather short, with slender labial palpi; ligula reaching thelevel of one third of palpomere I and distinctly bifid apically.THORAX. Pronotum transverse, wider than head, with very distinct notal carina, reaching the levelof seta PR2; legs normally developed, claws unequal, anterior twice as long as posterior in the firstpair.ABDOMEN. Urogomphi parallel and longer than width of tergum IX, with ancestral setae as long asurogomphi; anal tube long and slender, reaching the level between setae UR5 and UR6.Second and third instarsSame character states as in the first instar except for following:HABITUS. Width of head capsule 1.38–1.57 mm (x = 1.50 mm, n = 5) in the second and 1.84–1.98 mm(x = 1.90 mm, n = 6) in the third instar.MICROSCULPTURE. Granulated microsculpture also on postscuta of metanotum and abdominal terga,pointed microsculpture on urogomphi.CHAETOTAXY. Head: 2 additional setae on inner side of antennomere I, 4–6 setae on inner side ofantennomere II, 2–3 setae on outer side of stipes, 2 setae near base of mandible, 2–3 setae near102

Figs 11–19. Harpalus (Harpalus) cisteloides hurkai Divoký, Pulpán et Rébl. First instar larva: 11 – head capsule(dorsal view). 12 – nasale. 13 – mandible. 14 – maxilla. 15 – labium. Third instar larva: 16 – nasale. 17 – antenna. 18– mandible. 19 – urogomphi. Scales: 1 mm (Figs 11, 19), 0.5 mm (Figs 13, 14, 15, 17, 18), 0.2 mm (Figs 12, 16).103

apex of palpomere I, 5–7 pairs of setae (8–9 pairs in L3) on lateral side of prementum; 2 setae onplace of inner and outer stemmatal furrows, many diminutive setae or sensillae on dorsal surface ofhead capsule. Thorax: Pronotum with 2–3 additional setae along notal carina, number of setae onlateral margin and between setae PR11 and PR12, more than 8 additional spines on each side ofanterior femora. Abdomen: 1 additional seta between TE1 and TE6, 2 setae between TE6 and TE7,1–2 setae between TE9 and TE10 and 2–3 small setae on lateral margin. Length of URβ 0.75 timesthat of UR4, URγ about as long as UR4.HEAD. Nasale (Fig. 16) with larger lateral teeth; coronal suture longer than antennomere II; seta PA4as long as antennomeres II or III; setae PA1 – PA3 very small; antenna (Fig. 17); mandible (Fig. 18).THORAX. Legs long and slender, claws unequal as in first instar.ABDOMEN. Urogomphi (Fig. 19) subparallel and long, twice as long as width of tergum IX andlonger than anal tube.Harpalus (Harpalus) luteicornis (Duftschmid, 1812)(Figs 20–27)First instarHABITUS AND COLOR. Body yellow-white, retinaculum and apex of mandible brown. Width of headcapsule 1.04–1.14 mm (x = 1.08 mm, n = 5).MICROSCULPTURE. Isodiametric microsculpture on anterior part of frontale, granulated microsculptureon prescutum and postscutum of pronotum.CHAETOTAXY. Head: Setae PA4, PA5 and especially PA1 – PA3 very fine and small, setae PA6 andPA8 markedly fine and reduced; group gMX consists of 4–5 thicker and more than 50 thinner andrather short setae. Thorax: Except for ancestral chaetotaxy 6 or 7 additional spines on either side ofanterior femora.HEAD (Fig. 20) transverse (index width/length 1.3); nasale (Fig. 21) protruding, with fine but distinctcentral incision and 6–7 teeth on each side, two lateral slightly longer, adnasalia moderatelysloping, ventral row consists of 22–24 teeth; cervical grooves clearly bent, almost reaching thelevel of seta PA7 on dorsal side, don’t reaching the level of seta PA16 on ventral side; coronalsuture as long as width of antennomere I; egg-burster consists of 4–5 teeth, being larger caudad,along frontal suture on each side. Antennae slightly longer than mandibles; mandible (Fig. 22)strongly curved and robust, twice as long as basal width, with a toothlike swelling in front ofretinaculum, penicillus present; maxillae (Fig. 23) slender, stipes almost 3 times as long as wide;labium (Fig. 24) short, with slender labial palpi; ligula reaching the level of one fourth or fifth ofpalpomere I and weakly bifid at apex.THORAX. Pronotum narrower than head, with indistinct notal carina; legs normally developed,claws unequal, anterior as long as or a little longer than posterior.ABDOMEN. Abdominal tergites transverse; urogomphi weakly curved, twice as long as width oftergum IX and about one third of anal tube length.Third instarSame character states as in the first instar except for following:HABITUS. Width of head capsule 1.30–1.40 mm (x = 1.34 mm, n = 5).MICROSCULPTURE. Isodiametric microsculpture weakly distinct.CHAETOTAXY. Head: 2 additional setae on inner side of antennomere II, 2 setae on outer side ofstipes, 1–2 seta near apex of palpomere I, 7–9 pairs of setae on dorso-lateral surface of prementum;2–3 small setae on place of outer stemmatal furrows. Thorax: 1 (2) secondary setae on either side of104

Figs 20–27. Harpalus (Harpalus) luteicornis (Duftschmid). First instar larva: 20 – head capsule (dorsal view). 21– nasale. 22 – mandible. 23 – maxilla. 24 – labium. Third instar larva: 25 – nasale. 26 – antenna. 27 – mandible.Scales: 0.5 mm (Fig. 20), 0.3 mm (Figs 21, 22, 23, 24, 25, 26, 27).105

anterior femora. Only diminutive setae on dorsal surface of head capsule, thoracic and abdominaltergites. Length of seta URβ about 0.5 times, of URγ 0.75 times that of UR4.HEAD similar to L1, nasale (Fig. 25) more prominent and with larger lateral teeth; cervical grooves verydistinct and bent; well developed outer stemmatal furrows; antenna (Fig. 26), mandible (Fig. 27).THORAX. Pronotum with weakly developed notal carina; legs similar to L1; claws unequal, anteriorabout one fourth longer than posterior.ABDOMEN. Urogomphi slender and long, more than twice as long as width of tergum IX and lengthof anal tube.Harpalus (Harpalus) picipennis (Duftschmid, 1812)(Figs 28–35)First instarHABITUS AND COLOR. Small larvae with transverse, angular head capsule and shortened extremities;body yellow-white, head and pronotum yellow-brown, retinaculum and apex of mandibles darkbrown. Width of head capsule 0.70–0.78 mm (x = 0.73 mm, n = 6).MICROSCULPTURE. Isodiametric microsculpture absent, granulated microsculpture on pronotal prescutumand postscutum.CHAETOTAXY. Except for ancestral chaetotaxy 4 additional setae on either side of anterior femora;seta PA4 distinctly longer than setae PA1 – PA3 and PA5.HEAD. (Fig. 28) transverse (index width/length 1.4); nasale (Fig. 29) slightly prominent, with 7–9central and 2, slightly longer, lateral teeth on each side, ventral row consists of 19–23 teeth,adnasalia moderately sloping; cervical grooves bent, almost reaching the level of seta PA3; coronalsuture very short, as long as half of width of antennomere I; egg burster consists mostly of 2teeth, one larger on the level of seta PA4, one smaller on the level of seta PA3; group gMX consistsof only 1–2 thick and more than 40 thinner, very fine and short setae. Antennae short and thickset,less than 6.5 times as long as wide; mandibles (Fig. 30) strongly curved and robust, twice as longas basal width, with a fine swelling in front of retinaculum, penicillus present; maxillae (Fig. 31)thickset, stipes only 2.3 times as long as wide; labium (Fig. 32) short with slender labial palpi, ligulareaching the level of one third of palpomere I.THORAX. Pronotum transverse, with distinct notal carina; legs short and robust.ABDOMEN. Abdominal tergites transverse; urogomphi very short and divergent, only slightly longerthan width of tergum IX and length of anal tube.Second and third instarsSame character states as in the first instar except for following:HABITUS. Width of head capsule 0.83–0.97 mm (x = 0.92 mm, n = 3) in the second and 0.94–1.20 mm(x = 1.06 mm, n = 6) in the third instar.MICROSCULPTURE. Weakly distinct isodiametric microsculptur on head capsule, granulated microsculpturealso on postscuta of meso- and metanotum.CHAETOTAXY. Head: 2 additional setae on inner side of antennomere II, 2 setae on outer side ofstipes, 2 setae near apex of palpomere I, 5–6 pairs of setae (8–9 pairs in L3) on dorso-lateral surfaceof prementum; 1–2 small setae on place of inner and outer stemmatal furrows. Thorax: 6 additionalsetae on each side of anterior femora. Abdomen: On abdominal terga 1 additional seta between TE1and TE6, 2 setae between TE6 and TE7 and 1 seta between TE9 and TE10; 4 long secondary setaeon urogomphi. Several very small setae on surface of thoracic and abdominal terga and urogomphi.Length of both URβ and URγ about 0.80 times that of UR4.106

Figs 28–35. Harpalus (Harpalus) picipennis (Duftschmid). First instar larva: 28 – head capsule (dorsal view). 29– nasale. 30 – mandible. 31 – maxilla. 32 – labium. Third instar larva: 33 – nasale. 34 – antenna. 35 – mandible.Scales: 0.5 mm (Fig. 28), 0.2 mm (Figs 29, 30, 31, 32, 33, 34, 35).107

HEAD. Similar to L1; nasale (Fig. 33) more protruding; coronal suture longer; cervical groovesdeeper; outer stemmatal furrows weakly distinct. Extremities slightly longer: antenna (Fig. 34);mandible (Fig. 35).THORAX. Pronotum with weakly determined notal carina; legs a little longer.ABDOMEN. Urogomphi a little longer than width of tergum IX and the length of anal tube.Harpalus (Harpalus) saxicola Dejean, 1829(Figs 36–44)First instarHABITUS AND COLOR. Body pale, retinaculum and apex of mandible dark brown. Width of headcapsule 1.42 and 1.46 mm.MICROSCULPTURE. Isodiametric microsculpture absent, granulated microsculpture on prescurumand postscutum of pronotum and on postscuta of mesonotum, metanotum and of abdominaltergites I–III.CHAETOTAXY. Except for ancestral chaetotaxy one additional seta on dorso-lateral surface of prementumand 7 additional setae on either side of anterior femora; group gMX consists of 10–11thick and more than 60 thinner, rather short setae; seta PA4 very long, as long as antennomere I and3–4 times as long as seta PA5, setae PA1 – PA3 very short.HEAD (Fig. 36) transverse (index width/length 1.3); nasale (Fig. 37) protruding, almost reaching thetop of adnasalia, with 8–10 central teeth and two lateral on each side twice as long as the others,ventral row consists of 25–27 teeth; cervical grooves bent, reaching the level of seta PA5; coronalsuture as long as antennomere II; egg burster consists of 1 small teeth behind the level of seta PA7.Antennae as long as mandibles; mandible (Fig. 38) strongly curved and robust, 1.8 times as longas basal width, with only a fine swelling in front of retinaculum; maxilla (Fig. 39) with stipes 3 timesas long as wide; labium (Fig. 40) slender, ligula reaching the level of one fourth of palpomere I.THORAX. Pronotum transverse, wider than head, with well developed notal carina, reaching thelevel of seta PR2; legs (Fig. 41) short and robust, claws unequal, anterior distinctly shorter thanposterior.ABDOMEN. Abdominal terga transverse with ancestral setae; urogomphi very short and weaklydivergent, only a little longer than width of tergum IX and length of anal tube. Setae UR4–8 longerthan urogomphi, shifted apicad.Second and third instarSame character states as in the first instar except for following:HABITUS. Width of head capsule 1.57 mm in L2 and 1.95 and 1.98 mm in L3.MICROSCULPTURE. Isodiametric microsculpture on head and pronotum.CHAETOTAXY. 1 additional seta on inner side of antennomere I (2 setae in L3), 3 setae on inner sideof antennomere II, 1 long and 1 short setae on outer side of stipes (2 long and 2 short setae in L3),2 setae near base of mandible, 1 seta near apex of palpomere I (2 setae in L3), 6–10 pair of setae ondorso-lateral side of prementum; 2 setae on place of inner and outer stemmatal furrows, 1–2 setaenear very long seta PA4, 2 pairs of small setae on the level of seta FR4.Thorax: 2–3 secondary setaein both anterior and posterior rows, 5–7 setae on lateral margin, 2 setae near prothoracic notalcarina; 2 secondary spines and 2–3 setae on either side of anterior femora. Numerous very smallsetae on head capsule, thoracic and abdominal terga and urogomphi. Length of seta URβ about0.75 times that of UR4, URγ about as long as UR4.108

Figs 36–44. Harpalus (Harpalus) saxicola Dejean. First instar larva: 36–head capsule (dorsal view). 37 – nasale.38 – mandible. 39 – maxilla. 40 – labium. 41 – first leg. Third instar larva: 42 – nasale. 43 – antenna. 44 –mandible. Scales: 1 mm (Fig. 36), 0.5 mm (Figs 38, 39, 40, 41, 43, 44), 0.2 mm (Figs 37, 42).109

HEAD similar to L1; nasale (Fig. 42) more protruding, overlapping top of adnasalia, with 9–10 centralteeth, two lateral on each side very large, ventral row consists of about 26 teeth, adnasalia moderatelysloping; cervical grooves more bent; antennae slender (Fig. 43); mandible (Fig. 44).ABDOBEN. Urogomphi very short and divergent, shorter than width of tergum IX and only slightlylonger than anal tube.Harpalus servus (Harpalus) (Duftschmid, 1812)(Figs 45–53)First instarHABITUS AND COLOR. Body yellow-brown, head capsule, apex of mandibles, retinaculum, thoracicand abdominal sclerites dark brown. Width of head capsule 0.84–1.02 mm (x = 0.94 mm, n = 18).MICROSCULPTURE. Isodiametric microsculpture absent, granulated microsculpture on prescutumand postscutum of pronotum and of postscuta of mesonotum, metanotum and abdominal tergitesI–III.CHAETOTAXY. Except for ancestral chaetotaxy 2 additional spines on either side of anterior femora;seta PA4 relatively long, more than twice as long as PA5 and more longer than PA1–PA3; groupgMX consists of two or three thick and more than 50 thinner, rather short setae.HEAD (Fig.45) transverse (index width/length less than 1.3); nasale (Fig. 46) protruding and weaklybifurcate at apex, with 7–8 central teeth, two lateral teeth only a little longer; ventral row consistsof 16–18 teeth; cervical grooves clearly bent, almost reaching the level of seta PA5; coronal suturevery short, as long as length of setae PA1–PA3; egg burster consists on each side of only onelarger tooth. Extremities short and robust; antennae as long as mandibles; mandible (Fig. 47)strongly curved, twice as long as basal width, with one small swelling in front of retinaculum,penicillus present; maxilla (Fig. 48) stout and robust, stipes 2.4 times as long as wide; labium (Fig.49) short with ligula reaching the level of one third of palpomere I and slightly bifid at apex.THORAX. Pronotum distinctly transverse, wider than head, with distinct notal carina, reaching thelevel of seta PR14; legs (Fig. 50) robust, claws unequal, anterior twice as long as posterior in firstpair, anterior a little longer in other pairs.ABDOMEN. Urogomphi very short and curved, a little longer than width of tergum IX and slightlylonger than anal tube.Second and third instarsSame character states as in the first instar except for following:HABITUS. Width of head capsule 1.05–1.27 mm (x = 1.18 mm, n = 4) in the second and 1.50–1.64 mm(x = 1.59 mm, n = 6) in the third instar.MICROSCULPTURE. Isodiametric microsculpture on head and pronotum.CHAETOTAXY. Head: 2 additional setae on inner side of antennomere II, 2 setae on outer side ofstipes, 1–2 setae near apex of palpomere I, more than 10 pairs of setae on dorso-lateral surface ofprementum; 1 or 2 setae on places of inner and outer stemmatal furrows. Thorax: pronotum with 2secondary setae near notal carina and 4–5 setae on lateral and 2 setae on posterior margin; 6secondary spines on either side of anterior femora. Abdomen: 1 seta between TE1 and TE6, betweenTE6 and TE7 and between TE9 and TE10. Length of seta URβ about 0.75 that of UR4, seta URγabout as long as UR4.HEAD similar to L1; nasale (Fig. 51) more prominent and with larger lateral teeth; antenna (Fig. 52);mandible (Fig. 53); maxilla longer than in first instar, stipes more than 3.5 times as long as wide.THORAX. Legs with unequal tarsal claws.ABDOBEN. Urogomphi parallel, slightly longer than width of tergum IX and distinctly longer thananal tube.110

Figs 45–53. Harpalus (Harpalus) servus (Duftschmid). First instar larva: 45–head capsule (dorsal view). 46 –nasale. 47 – mandible. 48 – maxilla. 49 – labium. 50 – first leg. Third instar larva: 51 – nasale. 52 – antenna. 53– mandible. Scales: 0.5 mm (Fig. 45), 0.3 mm (Figs 47, 48, 49, 50, 52, 53), 0.2 mm (Figs 46, 51).111

Harpalus (Harpalus) solitaris Dejean, 1829(Figs 54–61)Third instarHABITUS AND COLOR. Body yellow-white, head and pronotum yellow-brown, retinaculum and apexof mandible dark brown. Width of head capsule 1.91 mm in one specimen.MICROSCULPTURE. Isodiametric microsculpture on parietale and pronotum, granulated microsculptureon prescutum and postscutum of pronotum and on postscuta of mesonotum, metanotum andabdominal terga I–III.CHAETOTAXY. Head: 2 additional setae on inner side of antennomere II, 2 setae on outer side ofstipes, 2 setae near apex of palpomere I, 7–11 pairs of setae on dorso-lateral surface of prementum;2–3 setae on place of inner and number of setae or sensillae on place of outer stemmatal furrows,one small seta along cervical grooves, group gMX consists of only two or three thicker and morethan 70 thinner, long setae; seta PA5 and especially PA1 – PA3 small, seta PA4 three times as longas seta PA5 and as long as antennomere I, the majority of setae on frontale, except for FR2 and FR7,relatively fine. Thorax: several small secondary setae on thoracic tergites; 9–10 spines on eachside of anterior femora. Abdomen: tergites transvere (Fig. 60) with 2 secondary setae neighbouringTE1 and TE10 and one seta neighbouring TE6. Length of seta URβ 0.6 times that of UR4, seta URγabout as long as UR4.HEAD (Fig. 54) distinctly transverse (index width/length 1.3–1.4); nasale (Fig. 55) slightly protruding,with only 5–6 large central teeth and two slightly larger lateral teeth on each side, ventral rowconsists of about 25 teeth; adnasalia strongly sloping; cervical grooves long and bent, reachingthe level of seta PA7 dorsally and the level of seta PA15 on ventral side; coronal suture slightlylonger than width of antennomere I. Antennae (Fig. 56) as long as mandibles; mandible (Fig. 57)triangular and robust, 1.9 times as long as basal width, with two distinct additional teeth in front ofretinaculum, penicillus present; maxillae (Fig. 58) slender, stipes almost 3.3 times as long as wide;labium (Fig. 59), ligula reaching the level of one fourth of palpomere I.THORAX. Pronotum transverse, notal carina weakly developed; legs normally developed, clawsunequal, anterior twice as long as posterior in first pair, a little longer in other pairs.ABDOMEN. Urogomphi (Fig. 61) relatively long and weakly curved, twice as long as width of tergumIX, about one third as long as length of slender anal tube.Harpalus (Harpalus) xanthopus winkleri Schauberger, 1923(Figs 62–70)First instarHABITUS AND COLOR. Body yellow-brown, head and pronotum pale brown, retinaculum and apex ofmandible dark brown. Width of head capsule 0.98 mm in one specimen.MICROSCULPTURE. Isodiametric microsculpture on head very distinct.CHAETOTAXY. Ancestral chaetotaxy on head capsule and extremities; seta PA4 and especially setaePA5, PA6 and PA8 short and reduced.HEAD distinctly transverse; nasale (Fig. 62) only slightly protruding, with 10 central teeth and twoa little larger lateral teeth on each side, ventral row consists of 25–26 teeth, adnasalia moderatelysloping; cervical grooves short and weakly bent, almost reaching the level of seta PA5; coronalsuture as long as width of antennomere I; egg burster consists of 5 small teeth along frontal sutureon each side. Extremities, especially antennae and mandibles, short and robust; mandible (Fig. 63)triangular, 1.9 times as long as basal width, with only one small tooth in front of retinaculum,112

Figs 54–61. Harpalus (Harpalus) solitaris Dejean. Third instar larva: 54–head capsule (dorsal view). 55–nasale.56–antenna. 57–mandible. 58–maxilla. 59–labium. 60–tergum I. 61–urogomphi. Scales: 1 mm (Figs 54, 60, 61),0.5 mm (Figs 56, 57, 58, 59), 0.3 mm (Fig. 55).113

penicillus present; maxilla slender, stipes more than 3 times as long as wide; labium short and ratherrobust.THORAX AND ABDOMEN. Not available.Third instarSame character states as in the first instar except for following:HABITUS. Width of head capsule 1.37 mm in one specimen.MICROSCULPTURE. Isodiametric microsculpture on head, thoracic and abdominal tergites. Granulatedmicrosculpture on pronotal prescutum and postscutum and on postscuta of mesonotum, metanotumand abdominal tergites I–II(III).CHAETOTAXY. Head: 2 secondary setae on inner side of antennomere II, 2 setae on outer side ofstipes, 1 seta near apex of labial palpomere I, 8 pairs of setae on dorso-lateral surface of prementum,4 setae near outer stemmatal furrow. Thorax: 2 small secondary setae along notal carina on prothorax;8 additional spines on either side of anterior femora. Abdomen: seta TE11 on abdominal tergavery reduced or rudimental. Length of seta URβ only about 0.2 times that of UR4 and about 0.3times that of URγ.HEAD (Fig. 64) transverse (index weight/length 1.4); nasale (Fig. 65) more prominent, with 9–10central teeth, two lateral on each side distinctly longer, ventral row consists of 25 teeth, adnasaliamore sloping; cervical grooves more deepened and bent; coronal suture longer than width ofantennomere I; outer stemmatal furrow developed. Extremities more slender; antenna (Fig. 66);mandible (Fig. 67) with one distinct additional tooth in front of retinaculum; maxilla (Fig. 68); labium(Fig. 69).THORAX. Legs with unequal claws, anterior distinctly shorter than posterior.ABDOMEN. Urogomphi (Fig. 70) long and parallel, 1.8 times as long as width of tergum IX and aboutone third longer than anal tube.LARVAL DIAGNOSIS OF SUBGENUS HARPALUSFirst instarHABITUS. Typical Harpalini larva with great and transverse head capsule and pronotum, body beingnarrower caudad.MICROSCULPTURE. Mostly only parts of frontale and parietale and lateral sides of pronotum withisodiametric microsculpture. Granulated microsculpture on pronotal prescutum and postscutumand on the postscuta of mesonotum, metanotum and of anterior abdominal tergites. Pointed microsculpturesometimes on tergite IX, on base of urogomphi and on antero-lateral parts of abdominalscuta.CHAETOTAXY. All ancestral setae and pores, except for seta LA4 on prementum, present; one additionalseta of ancestral origin on adnasalia, and mostly one additional pair of spiniform setae orspines on all pairs of femora.HEAD. Distinctly transverse (index width/length more than 1.1), nasale diverse but frequentlyprominent and always with central protuberance and one pair of lateral teeth on each side; cervicalgrooves and coronal suture always present; egg-bursters consist of 2 rows of 1–10 teeth alongfrontal suture. Antennae mostly a little longer than mandibles; mandible more or less incurvate andstout, as a rule twice as long as wide, frequently with only one or none tooth in front of retinaculum,exceptionally with three extra teeth, penicillus present; six stemmata well-developed; maxillarystipes different in length, 1.5 to 5 times as long as wide, lacinia distinct, with stout lateral seta MX6;prementum longer than wide, ligula at least as wide as base of palpomere I, frequently bifid at apex.114

Figs 62–70. Harpalus (Harpalus) xanthopus winkleri Schauberger. First instar larva: 62 – nasale. 63 – mandible.Third instar larva: 64 – head capsule (dorsal view). 65 – nasale. 66 – antenna. 67 – mandible. 68 – maxilla. 69 –labium. 70 – urogomphi. Scales: 1 mm (Figs 64, 70), 0.5 mm (Figs 63, 66, 67, 68, 69), 0.3 mm (Fig. 65), 0.2 mm(Fig. 62).115

THORAX. Pronotum transverse, mostly wider than head, often with well developed notal carinaabove half of pronotum; legs stout, spiny, claws frequently unequal.ABDOMEN. Abdominal tergites transverse, sometimes with a fine keel separating prescutum andscutum; urogomphi parallel or divergent, mostly longer than both width of tergite IX and length ofanal tube.Second and third instarsSame character states as in the first instar except the following:HABITUS. Head capsule wider than in first instar.MICROSCULPTURE. In some species isodiametric microsculpture also on abdominal tergites. Pointedmicrosculpture on posterior abdominal tergites and on base of urogomphi.CHAETOTAXY. Secondary chaetotaxy regularly represented by these setae: Head. 2–3 (1–6) setae oninner side of antennomere II, 2–3 setae on outer side of stipes, 2–3 (1) setae near apex of palpomereI, a number pairs of setae (5–6 pairs in L2, 8–9 pairs in L3 ) on dorso-lateral surface of prementum;2 setae on place of inner and outer stemmatal furrows; many diminutive setae or sensillae on dorsalsurface of head capsule. Thorax. Some setae around notal carina, on lateral border and betweensetae PR11 and PR12; legs often with many secondary setae. Abdomen. Abdominal tergites withseveral setae; urogomphi with 4 long secondary setae and often with many diminutive setae orsensillae on the surface; seta URα small or absent, seta URβ more or less shorter than setae URγand UR4.HEAD and nasale mostly similar to L1, nasale with rather larger lateral teeth and more distinct centralprotuberance. Cervical grooves and stemmatal furrows more deepened, extremities rather longer.THORAX AND ABDOMEN similar to L1, urogomphi regularly longer.DISCUSSIONThe fundamental paper on the taxonomy of the larvae of Harpalina was published by Brandmayr,Ferrero & Zetto Brandmayr, 1980. In this paper, they separated within the genus Harpalus Latreille,1802 sensu lato, the “ophonoid” and “harpaloid” lineages, based on distinctive, morphologicallarval characters. They also presented differential diagnoses and the key of all described taxawithin the genus group, and a list of more or less well described species, including the authors ofthe descriptions.The taxonomy of the genus Harpalus is far of being stabilised at present. Based on the combinationof both adult and larval characters we consider Ophonus Dejean, 1821, CryptophonusBrandmayr & Zetto Brandmayr, 1982, Pseudoophonus Motschulsky, 1844 and NipponoharpalusHabu, 1973 as generic level taxa.The last monographic treatments of Carabidae larvae (Arndt 1991, Luff 1993) offer morphologicaldata for 17 species (11 species respectively) within the genus Harpalus in our concept. Afteradding data from additional sources (Brandmayr et al. 1980, Gardner 1938, Habu 1973, Hrka 1975,1992, Kirk 1972, Puchkov 1992, Zeto Brandmayr & Brandmayr 1978), the number of species raisesto 32, of which 29 are in the subgenus Harpalus. In this paper we are adding larval descriptions offurther 8 species of the subgenus Harpalus. Most papers published so far are based on incompleteand insufficient material that does not fully take in consideration the variability of the larvalcharacters of the studied species. With the increased number of described taxa, it is becomingobvious that the variability of larval characters is considerably higher than previously thought. Itis also necessary to increase the spectrum of the characters used in larval taxonomy, particularlyby the characters concerning the chaetotaxy of the head and appendages.116

Based on the study of our larval material, as well as on data in the literature, the genus Harpalusin our concept may be characterised mainly by the following characters: (1) Nasale deepenedbetween adnasalia; (2) Labial palpomere I in both second and third instars with 2 (1–4) short setaenear apex; (3) Mandible with 0–4 teeth in front of retinaculum, mandibular apex never bifid; (4)Antennomere II in instars II and III with (1)2–5(6) large subapical setae directed inwards; (5) Innerside of stipes with a setal group gMX consisting of more than 40 setae.A slightly more than 10 subgenera are mentioned world-wide in the genus Harpalus in ourconcept (Lorenz 1998), several tens of species aggregates are recognised in the subgenus Harpalus(Kataev 1995). In addition to the subgenus Harpalus, some of the larval characters areavailable also for species of the subgenera Acardystus Reitter, 1908 (Habu 1973), Artabas DesGozis, 1882 (Puchkov 1992), Harpalophonus Ganglbauer, 1892 (Zeto Brandmayr & Brandmayr1978), Megapangus Casey, 1914 (Kirk 1972), Plectralidus Casey, 1914 (Kirk 1972) and SemiophonusSchauberger, 1933 (Hrka 1992). Taxonomic–morphological differences between the larvaeof the species aggregates of the subgenus Harpalus are often more significant than thosebetween the larvae of conventional subgenera.The results of our study of both larval and adult characters within the subgenus Harpalussensu novo mostly quite support the Kataev’s division in species aggregates that was establishedusing adult characters, as it can be documented by the following examples:Harpalus (Harpalus) atratusKataev 1995 established the separate species aggregate “atratus” for this species. This is fullysupported by the larval characters. Unique is the form of nasale (Figs 2, 7), the shape of mandiblewith two additional teeth in front of retinaculum (Fig. 3), the shape of first instar labium (Fig. 5), aswell as the general habitus of the larva.Harpalus (Harpalus) cisteloides hurkaiA separate “cisteloides” species aggregate was established for this and three additional speciesby Kataev 1995. This is fully supported by the larval morphology, particularly by the following:details in the development of the nasale, with the wide V–shaped medial emargination (Figs 12, 16),the development of the ancestral chaetotaxy, the minute setae PA2 and PA3, and particularly thereduced setae FR8, FR9 and an additional seta near them. Some of these characters are similar tothose of H. autumnalis from the next species aggregate, in particular the similar shape of themandible and of the egg–bursters, similar arrangement of the additional chaetotaxy of the labiumand of the legs (particularly of the femora), the similar development of the secondary chaetotaxy ofthe antennae, mandibles and of the maxillae. The actual relationships are not clear at present.Harpalus (Harpalus) picipennisA well defined “pumilus” species aggregate was established for this and seven additional species(= Actephilus Stephens, 1833). This is entirely supported by the larval morphology, especiallyafter a comparison with the larva of H. pumilus. The larvae of this group are characterised particularlyby the shape of the nasale (Figs 29, 33), by the markedly shortened head appendages, by thearrangement of the egg-bursters (Fig. 28), and by the shortened urogomphi. The shortened headappendages and urogomphi and the basic shape of the nasale, are similar to those of the “anxius”species aggregate, but in the latter the arrangement of the egg-bursters, the additional chaetotaxyof the legs and the details in the shape of the nasale are different.117

Harpalus (Harpalus) saxicolaBased on adult characters, the species was assigned, together with H. angulatus Putzeys and H.distinguendus (Duftschmid), to the “distinguendus” species aggregate. This taxonomical actionis fully supported by the larval morphology, particularly after a comparison with the larva of H.distinguendus. The species aggregate is characterised by the sagittiform nasale (Figs 37, 42), theshortened urogomphi, as well as by the secondary chaetotaxy of the antennae (Fig. 43), mandibles(Fig. 44) and the maxillae. Some of the characters, e.g. the somewhat similar shape of the nasale andthe secondary chaetotaxy may suggest a possible relationships to the “affinis” and “hospes”species aggregates.Harpalus (Harpalus) servusBased on adult characters, the species was assigned by Kataev 1995, together with 8 additionalspecies, to the “anxius” species aggregate. The larval morphology of H. anxius, H. servus and H.subcylindricus fully supports this action. The larvae of the named species are very difficult todistinguish. The species aggregate is mainly defined by the shape of the nasale with a small apicalemargination (Figs 46, 51), by the arrangement of the egg-bursters (Fig. 45), the shortened headappendages and urogomphi, and by the chaetotaxy of the legs (namely of the femora). Some of thecharacters are shared with the “pumilus” species aggregate (see above).The “latus” species aggregateKataev assigned 10 Palaearctic species to this species aggregate. About half of the species aredescribed in the larval stage in more or less details: H. latus, H. luteicornis, H. marginellus, H.solitaris and H. xanthopus winkleri. The larval features of this five species indicate that thespecies aggregate is likely not homogeneous. Harpalus luteicornis and H. xanthopus winkleriare related. Harpalus latus and H. solitaris differ by the shape of the mandible and nasale, thelarva of H. latus in addition by the conspicuously transverse head. The arrangement of the eggburstersis similar in all species examined. The species aggregate requires a taxonomic revisionbased on sufficient material of all larval instars.The comparison of both larval and adult characters of species of the genus Harpalus indicatesthat it is at present more proper to assign the related species to the species aggregates rather thanto separate subgenera. However, some of the species aggregates established by Kataev 1995should be re-examined.A c k n o w l e d g e m e n tThe senior author acknowledges the support of this study by the Research Project of the Ministry of Educationof the Czech Republic No. J 13/98113130004.REFERENCESARNDT E. 1991: Familie Carabidae. Pp.: 45–141. In: KLAUSNITZER B. (ed.): Die Larven der Käfer Mitteleuropas 1.Band 1 Adephaga. Krefeld: Goecke & Evers, 237 pp.BOUSQUET Y. 1985: Morphologie comparée des larves de Pterostichini (Coleoptera: Carabidae): Descriptions ettables de détermination des espces du nord-est de l’Amérique du nord. Natur. Can. (Rev. Écol. Syst.) 112:191–251.BOUSQUET Y. & GOULET H. 1984: Notation of primary setae and pores on larvae of Carabidae (Coleoptera:Adephaga). Can. J. Zool. 62: 573–588.BRANDMAYR P., FERRERO E. & ZETTO BRANDMAYR T. 1980: Larval versus imaginal taxonomy and the systematicstatus of the ground beetle taxa Harpalus and Ophonus (Coleoptera, Carabidae: Harpalini). Entomol. Gener. 6:335–353.CHU H. F. 1945: The Larvae of Harpalinae Unisetosae (Coleoptera, Carabidae). Entomol. Amer. 25: 1–71.118

EMDEN F. I. van 1942: A key to the genera of larval Carabidae (Coleoptera). Trans. R. Entomol. Soc. Lond. 92:1–99.GARDNER J. C. M. 1938: Immature stages of Indian Coleoptera (23) (Carabidae-contd.). Indian Forest Rec. 3:149–157, Plate I, II.HABU A. 1973: Fauna Japonica. Carabidae: Harpalini (Insecta: Coleoptera). Tokyo: Keigaku Publ. Co., 430 pp.Habu A. & Sadanaga K. 1963: Illustrations for Identifications of Larvae of the Carabidae Found in CultivatedFields and Paddy-fields (II). Bull. Natl. Inst. Agr. Sci. Ser. C 16: 151–157.HABU A. & SADANAGA K. 1965: Illustrations for Identification of Larvae of the Carabidae Found in CultivatedFields and Paddy-fields (III). Bull. Natl. Inst. Agr. Sci. Ser. C 19: 81–216.HABU A. & SADANAGA K. 1970a: Descriptions of some larvae of the Carabidae found in cultivated fields and paddyfields(I). Konty 38: 9–23.HABU A. & SADANAGA K. 1970b: Descriptions of some larvae of the Carabidae found in cultivated fields and paddyfields(II). Konty 38: 24–41.HRKA K. 1975: Larval diagnosis of the tribe Stenolophini and notes on the classification of the subfamilyHarpalinae (Coleoptera, Carabidae). Acta Entomol. Bohemoslov. 77: 247–256.HRKA K. 1992: The taxonomic status of Semiophonus (Coleoptera: Carabidae, Harpalini) and description of thelarva of Harpalus (Semiophonus) signaticornis. Acta Entomol. Bohemoslov. 89: 29–34.HRKA K. 1996: Carabidae of the Czech and Slovak Republics. Zlín: Kabourek, 565 pp.KATAEV B. M. 1995: Subtribe Harpalina. Pp.: 139–153. In: KRYZHANOVSKIJ O. L. et al.: A Checklist of the Ground-Beetles of Russia and Adjacent Lands (Insecta, Coleoptera, Carabidae). Sofia-Moscow: PENSOFT Publishers,271 pp.KIRK V. M. 1972: Identification of ground beetle larvae found in cropland in South Dakota. Ann. Entomol. Soc.Amer. 65: 1349–1356.LARSSON S. G. 1941: Larver [Larvae]. Pp.: 243–360. In: HANSEN V.: Biller XI. Sandspringere og Lobebiller(Cicindelidae og Carabidae) [Coleoptera XI. Cicindelidae and Carabidae]. Danmarks Fauna 47: 1–360.LARSSON S. G. 1968: Lobebillernes larver [Larvae of Carabidae]. Pp.: 282–433. In: HANSEN V.: Biller XXIVSandspringere og Lobebiller (Cicindelidae og Carabidae) [Coleoptera XXIV Cicindelidae and Carabidae].Danmarks Fauna 76: 1–451.LORENZ W. 1998: Systematic list of extant ground beetles of the world. Tutzing: W. Lorenz, 502 pp.LUFF M. L. 1993: The Carabidae (Coleoptera) larvae of Fennoscandia and Denmark. Fauna Entomol. Scandinavica27: 1–176.PRUNER L. & MÍKA P. 1996: List of settlements in the Czech Republic with associated map field codes for faunisticgrid mapping system. Klapalekiana 32 (Suppl.): 1–115 (In Czech, Engl. Summary).PUCHKOV A. V. 1992: The larva of Harpalus (Artabas) splendens (Coleoptera, Carabidae). Vest. Zool. 1992: 70–73(In Russian, Engl. Summary).SCHIDTE J. C. 1867: De metamorphosi eleutheratorum observationes. Naturh. Tidssk. 4: 439–552, Taf. XII–XII.SHAROVA I. C. 1958: Lichinki zhukov zhuzhelic (Carabidae) poleznykh i vrednykh v selskom khozyaistve [Thelarvae of Carabidae, beneficial and noxious to agriculture]. Uchen. Zap. Mosk. Pedag. Inst. 124: 4–165 (InRussian).SHAROVA I. C. 1964: Carabidae. Pp.: 112–195. In: GHILAROV M. S. (ed.): Opredelitel obitajushchikh v pochvelichinok nasekomykh [Key for the identification of soil inhabiting insect larvae]. Moskva: Nauka, 919 pp (InRussian).ZETTO BRANDMAYR T. & BRANDMAYR P. 1978: Morfologia pre-imaginale e note bionomiche su Harpalus(Harpalophonus) circumpunctatus italus Schaum (Coleoptera, Carabidae). Boll. Ist. Entomol. Univ. Bologna34: 65–74.119

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Acta Soc. Zool. Bohem. 66: 121–140, 2002ISSN 1211-376XObservations on seasonal changes in the occurrence and maturation of fivehelminth species in the pimelodid catfish, Rhamdia guatemalensis,in the cenote (= sinkhole) Ixin-há, Yucatán, MexicoFrantišek MORAVEC 1, 2) , Edgar MENDOZA-FRANCO 2) , Clara VIVAS-RODRÍGUEZ 2) ,Joaquin VARGAS-VÁZQUEZ 2) & David GONZÁLEZ-SOLÍS 2)1)Institute of Parasitology, Academy of Sciences of the Czech Republic, Branišovská 31,CZ–370 05 eské Budjovice, Czech Republic2)Centre for Research and Advanced Studies, National Polytechnic Institute (CINVESTAV-IPN), Mérida Unit,Carretera Antigua a Progreso Km 6, A. P. 73 “Cordemex”, C. P. 973 10 Mérida, Yucatán, MexicoReceived March 23, 2001; accepted October 16, 2001Published June 28, 2002Abstract. Preliminary data on seasonal changes in the occurrence and the maturation of five fish helminths,the trematodes Genarchella tropica (Manter, 1936) and Stunkardiella minima (Stunkard, 1938), thecestode Proteocephalus brooksi García-Prieto, Rodríguez et Pérez-Ponce de León, 1996 and the nematodesRhabdochona kidderi Pearse, 1936 and Neophilometroides caudatus (Moravec, Scholz et Vivas-Rodríguez,1995), in their definitive host, Rhamdia guatemalensis (Günther, 1864), are provided, based on monthlysamples of fish (a total of 82 fish were examined) collected from the cenote (= sinkhole) Ixin-há, centralState of Yucatán, Mexico, from June – November 1994. Encysted metacercariae of Stunkardiella minimaand encapsulated larvae of Proteocephalus brooksi also occurred in Rhamdia guatemalensis. No clear-cutseasonal maturation cycles were observed in these parasites and in contrast to most helminths of freshwaterfish in the temperate zone, all these species seemed to reproduce throughout the year. High water temperaturesand quickly developing sequence of generations of invertebrate intermediate hosts appear to be the principlefactors determining this annual reproduction of fish helminths in the tropics.Seasonality, maturation cycles, fish helminths, Pimelodidae, Rhamdia guatemalensis, MexicoINTRODUCTIONIt is well-known that most helminth parasites of fish live for only a short time and in the temperatezone, in addition to the species capable of reproduction in any season, there are others that growand mature only in that period of the year when favourable climatic and other conditions occur(Chubb 1979, 1982, Moravec 1994). In spite of both the theoretical and practical importance of suchdata, the seasonal maturation cycles have been studied in few species and almost exclusively inEurope and North America. In subtropical and tropical regions such studies have not been madeand, as far as the authors know, the only published data is on the seasonal occurrence and maturationof the nematodes Rhabdochona zacconis Yamaguti, 1935 in Tribolodon hakuensis (Günther,1877) in a subtropical region of Japan (Moravec et al. 1998), of Procamallanus inopinatus Travassos,Artigas et Pereira, 1928 in Serrasalmus spilopleura Kner, 1858 in a subtropical region ofnortheastern Argentina (Hamann 1999), and of Rhabdochona kidderi in Cichlasoma nigrofasciatum(Günther, 1867) in a tropical region of central Mexico (Caspeta-Mandujano et al. 2000). Becausewater temperature is generally considered to be the principal factor controlling directly orindirectly the seasonal cycles of helminths of fish (Kennedy 1970), data on the seasonal maturationcycles of fish helminths in tropical regions are of special interest.121

Tab. 1. Monthly survey of Rhamdia guatemalensis (Günther) from the cenote Ixin-há for infection with Genarchellatropica (Manter)month no. of fish fish length mean no. of fish prevalence (%) intensity meanexamined (range) [cm] infected (range)June 20 18 (12–28) 8 40 1.6 (1–3)July 1 7 16 (12–23) 9 53 2.1 (1–4)August 14 19 (12–27) 5 36 1.6 (1–2)September 14 17 (13–19) 5 36 1.4 (1–2)October 15 16 (12–24) 2 13 1.5 (1–2)November 2 14 (11–17) 1 50 1.0 (1)The authors are aware that the material used in this work is rarther limited and that futher studies,covering all months of the year, are necessary to describe in detail the seasonal maturation cycles ofthese helminth species. Nevertheless, the present data represent, except for Rhabdochona kidderi,the first observations on the maturation cycles of these parasites in the tropical region (Yucatán),and therefore worthy of publication.MATERIALS AND METHODSThe cenote (= sinkhole) Ixin-há belongs to the so called open type (its opening is almost the same diameter as thediameter of the water surface) according to Hall (1977) and is situated in the central part of the Yucatán State(20° 37’ 14” N, 89° 06’ 40” W), southeastern Mexico. This is a deep water body of about 50 m in diametercommunicating with subterranean streams and surrounded by rich tropical vegetation. As in other parts of theYucatán Peninsula, the tropical setting, low elevation and strong maritime influences, the mean temperatures arewarm and relatively homogenous, with a mean annual temperature of 25–26 °C, and annual range of mean monthlytemperatures of about 4–6 °C (Lee 1996).The fish fauna of this cenote is insufficiently known but is dominated by the pimelodid catfish Rhamdiaguatemalensis (Günther, 1864) (Siluriformes: Pimelodidae); this fish is known to migrate from one cenote toanother through the interconnecting subterranean streams. Although no other species of fish were recordedduring this study, the rare occurrence of the two blind cave fishes, Ogilbia pearsei (Hubbs, 1938) (Ophidiiformes:Bythitidae) and Ophisternon infernale (Hubbs, 1938) (Synbranchiformes: Synbranchidae), cannot be excluded.Fish (Rhamdia guatemalensis) were caught by angling at regular monthly intervals from June until November1994 (Tab. 1). The live fish were transported to the laboratory of the CINVESTAV in Mérida, where they wereexamined for parasites. Altogether 82 specimens of R. guatemalensis were dissected. Occasional small samples ofbenthic invertebrates were examined for helminth larvae in June, July and August. This included aquatic Oligochaeta(17 specimens), Hirudinea (9), Gastropoda (12), Isopoda (1), Ephemeroptera (166), Plecoptera (2), Odonata(66), Trichoptera (19), Coleoptera (10), and Diptera (Chironomidae) (197).The freshly collected nematodes were fixed either in hot 4% formaldehyde (Rhabdochona kidderi) or hot 4%formaldehyde in physiological saline (Neophilometroides caudatus) and temporarily stored in 4% formaldehyde.For examination they were cleared in glycerine. After examination they were preserved in 70% ethanol. Trematodesand cestodes were fixed in 4% formaldehyde under a coverslip; subsequently they were stained with Schuberg’scarmine, dehydrated and mounted as permanent preparations in Canada balsam. The specimens have been depositedin the Institute of Parasitology, ASCR, in eské Budjovice and in the Parasitological Laboratory, CIN-VESTAV-IPN, in Mérida.RESULTSDuring this study, carried out at the cenote Ixin-há from June to November 1994, the following 13helminth parasites were recorded from Rhamdia guatemalensis: Trematoda: Genarchella tropica,Stunkardiella minima adults and metacercariae, Clinostomum cf. complanatum metacercariae;Cestoda: Proteocephalus brooksi adults and encapsulated larvae (= Nomimoscolex sp. and Proteocephalideagen. sp. larvae of Scholz et al. 1996), Dendrouterina pilherodiae Mahon, 1956 larvae122

and D. papillifera (Fuhrmann, 1908) larvae; Nematoda: Paracapillaria rhamdiae Moravec,González-Solís et Vargas-Vázquez, 1995, Pseudocapillaria yucatanensis Moravec, Scholz et Vivas-Rodríguez,1995, Neophilometroides caudatus (Moravec, Scholz et Vivas-Rodríguez, 1995),Rhabdochona kidderi, Contracaecum sp. Type 1 larvae and Eustrongylides sp. (cf. E. ignotusJägerskiöld, 1909) larvae; these findings were reported in the surveys of Yucatanese fish helminthspublished by Moravec et al. (1995a, b) and Scholz et al. (1995a, b, 1996).Of the above mentioned parasites, seasonal maturation was followed only in the species occurringas adults in Rhamdia guatemalensis, with the exception of the two capillariid species, Paracapillariarhamdiae and Pseudocapillaria yucatanensis, which only rarely occur in this host.The fish host and its foodIn this locality, the only definitive host of all five helminth species was the pimelodid catfish,Rhamdia guatemalensis. In the case of Rhabdochona kidderi, it cannot be excluded that the blindcave fish Ogilbia pearsei is also a host, but the importance of this fish for the local population ofRhabdochona kidderi is negligible.Since the composition of a helminth fauna (both qualitatively and quantitatively) is considerablyaffected by the diet of the host, the character of the food of R. guatemalensis was also recordedmonthly. This revealed that there were almost no changes in the composition of food over theperiod of this study, which consisted mainly of larvae of aquatic insects (largely Ephemeroptera,Plecoptera and Odonata) and other aquatic invertebrates (mainly snails), and occasionally a largeproportion of plant material; tree flowers or bird feathers were found in the stomachs and it is highlyprobable that these fish fed also on bird and bat excrement (bats occur in huge numbers in thislocality). Although no fish remains were recorded, the frequent occurrence of encapsulated Proteocephalusbrooksi larvae and encysted metacercariae of Stunkardiella minima suggests thatcannibalism frequently occurs in this species of fish in this eutrophic cenote.Genarchella tropica (Manter, 1936)This trematode species is a stomach parasite of pimelodid catfishes of the genus Rhamdia Bleeker,1858 in Mexico (Yucatán), Nicaragua and Panama (Scholz et al. 1995a, c). I was previously collectedfrom R. guatemalensis in the cenote Ixin-há by Scholz et al. (1995a).Occurrence of Genarchella tropica in Rhamdia guatemalensisPrevalence and intensity of infection: Out of 82 R. guatemalensis examined, 29 (prevalence 35%)were infected with G. tropica at an intensity of 1–4 (mean 2) trematodes per fish (see Tab. 1). Thetrematodes were all found in the host’s stomach.The number of catfish examined during the survey and the incidence of infection with G. tropicais given in Tab. 1. As is evident from Tab. 2, infection by this trematode is associated with the bodysize (age) of the fish. The smallest catfish that harboured G. tropica measured 12 cm and the largest23 cm. The trematodes occurred in all size-groups of catfish examined, which was all but the smallestsize-group. Tab. 2 shows that the values of both prevalence and mean intensity of infection generallydecreased with the body length of the host; a considerable decrease in prevalence is obviousin the largest size-group (body length 20–28 cm).Seasonal changes in prevalence and mean intensity of infectionA survey of prevalence, intensity and mean intensity of G. tropica infection in catfish in individualmonths is shown in Tab. 1. G. tropica was present in catfish during the whole period. It is obviousfrom Fig. 1 that the prevalence was usually rather high, attaining its maximum (53%) in July, with a123

Tab. 2. Association of helminth infection with body length of host (irrespective of season) (prevalence, meanintensity)helminth species > 15 cm (n = 31) > 19 cm (n = 36) < 20 cm (n = 15)Genarchella tropica 45% 39% 13%1.9 1.6 1.5Stukardiella minima 29% 19% 20%1.9 19.3 8.3Stunkardiella minimametacercariae 13% 11% 27%37.3 15.0 5.3Proteocephalus brooksi 6% 8% 13%1.0 5.3 5.5Proteocephalus brooksiencapsulated larvae 52% 44% 7%39.8 23.3 6.0Philometroides caudata 0% 14% 13%0 1.4 2.0Rhabdochona kidderi 87% 81% 67%10.1 6.6 8.2marked decrease (13%) in October; the mean intensity was relatively low throughout the period,with the highest value (2.1) in July.Seasonal changes in maturation of Genarchella tropicaThe state of maturation of G. tropica in the monthly samples is shown in Fig. 2. The first group oftrematodes included young individuals without eggs, the second: small individuals (body lengthFig. 1. Variation of prevalence (triangle) and mean intensity of infection (square) by Genarchella tropica (Manter)in Rhamdia guatemalensis (Günther) from the cenote Ixin-há over the period June to November 1994.124

about 1 mm) with only a few eggs in their uterus, and the third: large specimens (body length 1.3–2.2 mm) with many eggs in their uterus.It is evident from Fig. 2 that G. tropica had eggs in all months, but that there were distinctseasonal differences in the composition of trematode samples from individual months. Although inJune specimens with many eggs (group III) prevailed and some with few eggs (group II) werepresent, only specimens with many eggs occurred in July; in the period from August to October theproportion of specimens with many eggs gradually decreased and that of those with few eggsgradually increased; also young, nongravid individuals were recorded in August and October.Only trematodes with few eggs were found in November, but this monthly sample consisted of onlytwo individuals.Monthly changes in the number of trematode size groups are apparent from Fig. 3, which showsthe mean numbers of individuals in each size group of G. tropica in each month. It is obvious fromthis figure that there was a sudden increase in the numbers of trematodes with many eggs in July,reaching the maximum number of 1.24 specimens per fish; from August, the numbers of individualsin this group gradually decreased to zero by November. On the other hand, the numbers of trematodeswith few eggs increased gradually from zero in July to 0.50 in October–November. Numbersof nongravid specimens per fish, in August and October, were low (0.07 and 0.10).Fig. 2. Monthly changes in the occurrence and state of maturity of Genarchella tropica (Manter) in Rhamdiaguatemalensis (Günther) from the cenote Ixin-há over the period June to November 1994. The data are expressedas percentages of the total number of trematodes found per month: small specimens without eggs – group I(unshaded), specimens with few eggs – group II (stippled) and fully mature specimens – group III (blackened).125

Tab. 3. Survey of Rhamdia guatemalensis examined from the cenote Ixin-há and their infection with Stunkardiellaminimamonth no. of fish examined no. of fish infected prevalence (%) intensity mean (range)June 20 8 40 11.0 (1–31)July 1 7 3 1 8 1.0 (1)August 14 3 21 6.3 (2–11)September 14 3 21 19.0 (1–54)October 15 2 13 7.7 (1–26)November 2 0 – –Stunkardiella minima (Stunkard, 1938)This acanthostomatid trematode is a specific intestinal parasite of Rhamdia guatemalensis in southernMexico (Pérez-Ponce de León et al. 1996); conspecific metacercariae occur mostly in fins of R.guatemalensis and some other fishes. From the cenote Ixin-há, both adults and metacercariae ofthis species were reported by Scholz et al. (1995a, b).Occurrence of adult Stunkardiella minima in Rhamdia guatemalensisPrevalence and intensity of infection: Out of 82 R. guatemalensis examined from this locality, 19(prevalence 23%) proved to be infected with S. minima, and the intensity of infection was 1–54(mean 9) trematodes per fish (Tab. 3). The trematodes were found only in the host’s intestine.Fig. 3. Monthly changes in mean numbers of specimens of three size groups of Genarchella tropica (Manter) perfish. A = small specimens without eggs (group I); B = specimens with few eggs (group II); C = fully maturespecimens with numerous eggs (group III).126

The number of catfish examined each month and the incidence of infection with S. minima isgiven in Tab. 3. They occurred in all size-groups of catfish, but it is evident from Tab. 2 thatinfection by this trematode is associated with the body size (age) of the fish; the highest prevalence(29%) and the lowest mean intensity (1.9) was in the smallest fish; the lowest prevalence and thehighest mean intensity was found in fish with a body length 16–19 cm. The smallest catfish harbouringS. minima measured 12 cm and the largest 28 cm.Seasonal changes in prevalence and mean intensity of infectionA survey of the monthly prevalence, intensity and mean intensity of S. minima infection of catfishis shown in Tab. 3. This trematode was found in catfish from June to October. The highest prevalence(40%) was in June, decreased (18–21%) in July-September, and again (13%) in October tozero in November, when only two fish were examined (Fig. 4). The mean intensity decreased (1) inJuly and then increased to attain its maximum (19) in September, followed by a decrease (13) inOctober.Seasonal changes in maturation of Stunkardiella minimaThe state of maturation of S. minima in each month is shown in Fig. 5. The first group of trematodesincludes young individuals (body length 0.4–0.7 mm) without eggs, the second group – smallindividuals (body length 0.7–0.8 mm) with only a few eggs in their uterus, and the third group –larger specimens (body length 1.3–2.2 mm) with many eggs.Fig. 4. Variation in the prevalence (triangle) and mean intensity of infection (square) of Stunkardiella minima(Stunkard) of Rhamdia guatemalensis (Günther) from the cenote Ixin-há over the period June to November1994.127

Tab. 4. Monthly survey of Rhamdia guatemalensis (Günther) from the cenote Ixin-há for infection with metacercariaeof Stunkardiella minima (Stunkard)month no. of fish examined no. of fish infected prevalence (%) intensity mean (range)June 20 2 10 60.5 (1–120)July 1 7 4 2 4 6.8 (1–15)August 14 3 21 6.3 (2–8)September 14 1 7 43.0 (43)October 15 3 20 8.0 (4–16)November 2 0 – –It is evident from Fig. 5 that S. minima containing eggs were present in all months exceptNovember, when only two fish were examined and no S. minima was found. The specimens withmany eggs (group III) made up 100% of the samples in September and October and prevailed inthose collected in June–August, in which trematode groups I and II were present. The proportionof specimens without eggs (group I) gradually increased from June to August, whereas of thosewith few eggs (group II) was somewhat greater in July compared to June and August.Monthly changes in the number of trematode groups are apparent from Fig. 6, which shows themean numbers of individuals in each of the size group of S. minima in each month. The highestFig. 5. Monthly changes in the occurrence and state of maturity of Stunkardiella minima (Stunkard) in Rhamdiaguatemalensis (Günther) from the cenote Ixin-há over the period June to November 1994. The data are expressedas percentages of the total number of trematodes found per month: small specimens without eggs – group I(unshaded), specimens with few eggs – group II (stippled) and fully mature specimens with numerous eggs – groupIII (blackened).128

number (3.65) of trematodes with many eggs (group III) per fish occurred in June, then suddenlydecreased (0.35–0.57) in July–August and considerably increased (2.86) in September, and suddenlydecreased (0.57) in October. The numbers of trematodes in groups I and II were rather low inJune–August.Occurrence of Stunkardiella minima metacercariae in Rhamdia guatemalensisOut of 82 R. guatemalensis, 13 (prevalence 16%) harboured S. minima metacercariae encysted intheir fins, with the intensity of infection being 1–120 (mean 18) cysts per fish.The number of catfish examined each month and their infection with encysted metacercariae ofS. minima is given in Tab. 4. The highest prevalence was in July–August and October, and thehighest intensity in June and September. Tab. 2 shows that the mean intensity decreased withincrease in size of fish, but the highest prevalence was in the largest fish. The data suggest anoverdispersion in the frequency distribution of infections.Proteocephalus brooksi García-Prieto, Rodríguez et Pérez-Ponce de León, 1996Adults of this cestode are specific intestinal parasites of Rhamdia guatemalensis in southernMexico (Pérez-Ponce de León et al. 1996). In the cenote Ixin-há, the adults of this species wereFig. 6. Monthly changes in mean numbers of specimens of three size groups of Stunkardiella minima (Stunkard)per fish. A = small specimens without eggs (group I); B = specimens with few eggs (group II); C = fully maturespecimens with numerous eggs (group III).129

Tab. 5. Monthly survey of Rhamdia guatemalensis (Günther) from the cenote Ixin-há for infection with Proteocephalusbrooksi García-Prieto et al. (excluding encapsulated larvae)month no. of fish examined no. of fish infected prevalence (%) intensity mean (range)June 20 3 15 3.0 (1–6)July 1 7 1 6 1.0 (1)August 14 2 14 5.5 (3–8)September 14 1 7 1.0 (1)October 15 0 – –November 2 0 – –identified as Nomimoscolex sp. by Scholz et al. (1996). The encapsulated cestode larvae occurringin R. guatemalensis in this locality, designated as Proteocephalidea gen. sp. larvae by Scholz et al.(1996), undoubtedly belong to the same species, because no other proteocephalids were found inthis locality.Occurrence of adult Proteocephalus brooksi in Rhamdia guatemalensisPrevalence and intensity of infection: Out of 82 R. guatemalensis from this locality, only 7 (prevalence9%) harboured P. brooksi in their intestines, the intensity of infection was 1–13 (mean 4)cestodes per fish.The number of catfish examined and their infection with P. brooksi is given in Tab. 5; it is evidentthat this parasite was found only from June to September. Tab. 2 shows that infection by thisFig. 7. Variation in the prevalence (diamond) and mean intensity of infection (sguare) by Rhabdochona kidderiPearse in Rhamdia guatemalensis (Günther) from the cenote Ixin-há over the period June to November 1994.130

cestode was distinctly associated with the body size of the host, and both the prevalence and theintensity increased with increasing body size of fish. Because of the low rate of infection it was notpossible to quantify the seasonal changes in prevalence and mean intensity.Seasonal changes in maturation of Proteocephalus brooksiBecause of the rarity of P. brooksi in the intestine of catfish, it was impossible to make a detailedquantitative analysis of its state of maturity each month. The following specimens were found:June: 2 gravid specimens (body length about 4 cm), 4 nongravid specimens with mature segments(1.5–3 cm) and 10 immature specimens (1–1.5 cm); July: 1 immature specimen; August: 2 gravidFig. 8. Monthly changes in the occurrence and state of maturity of Rhabdochona kidderi Pearse in Rhamdiaguatemalensis (Günther) from the cenote Ixin-há over the period June to November 1994. The data are expressedas percentages of the total number of nematodes found per month: larvae and females without eggs (unsheaded),males (stippled), females with immature eggs in uteri (obliquely hatched) and gravid females with mature eggs inuteri (blackened).131

Tab. 6. Monthly survey of Rhamdia guatemalensis (Günther) from the cenote Ixin-há for infection with encapsulatedlarvae of Proteocephalus brooksi García-Prieto et al.month no. of fish examined no. of fish infected prevalence (%) intensity mean (range)June 20 9 45 30.0 (6–79)July 1 7 7 4 1 23.0 (1–45)August 14 6 43 21.0 (3–165)September 14 4 29 10.5 (4–22)October 15 7 47 32.7 (6–72)November 2 0 – –specimens, 2 nongravid specimens with mature segments and 5 immature specimens; September: 1larva (scolex) (length about 1 mm). This suggests that oviposition occurred in June and August,and new infections were acquired by fish from June to September.Occurrence of encapsulated Proteocephalus brooksi larvae in Rhamdia guatemalensisOut of 82 R. guatemalensis, 33 (prevalence 40%) had P. brooksi larvae encapsulated in theirmesenteries, the intensity of infection being 1–165 (mean 30) larvae per fish.The number of catfish examined and their infection with encapsulated P. brooksi larvae is givenin Tab. 6. Larvae were found from June to October with high values of prevalence and meanintensity, except in September, when both values were distinctly low. No larvae were found inNovember, but this may be due to the fact that only two catfish were examined. Tab. 2 shows thatthe rate of infection is associated with the body size (age) of the fish. Values for both the prevalenceand mean intensity decreased with increase in body length of the catfish.Neophilometroides caudatus (Moravec, Scholz et Vivas-Rodríguez, 1995)This nematode is a specific parasite of the swimbladder surface of R. guatemalensis, so far onlyreported from southeastern Mexico from the cenotes Ixin-há and Xmucuy in Yucatán and thePapaloapan River in Veracruz (Moravec et al. 1995a, c, 2002).Occurrence of Neophilometroides caudatus in Rhamdia guatemalensisPrevalence and intensity of infection: Of the 82 R. guatemalensis from this locality, 7 (prevalence9%) were infected with N. caudatus, the intensity of infection being 1–2 (mean 2) nematodes perfish. All nematodes were located under the serosa of the host’s swimbladder.The number of catfish examined and their infection with N. caudatus is given in Tab. 7. Thisparasite was recorded from June to October. Because of its rarity, it was impossible to compare dataTab. 7. Monthly survey of Rhamdia guatemalensis (Günther) from the cenote Ixin-há for infection with Neophilometroidescaudatus (Moravec et al.)month no. of fish examined no. of fish infected prevalence (%) intensity mean (range)June 20 2 10 1.5 (1–2)July 1 7 1 6 2.0 (2)August 14 1 7 2.0 (2)September 14 1 7 1.0 (1)October 15 2 13 1.5 (1–2)November 2 0 – –132

from individual months. Tab. 2 shows that N. caudatus infections occurred only in large catfish,i. e., longer than 15 cm.Seasonal changes in maturation of Neophilometroides caudatusBecause of the rarity of N. caudatus, it was impossible to make a detailed quantitative analysis ofthe state of maturity of this parasite each month. The following nematodes were found: June: 3nongravid females; July: 2 nongravid females; August: 1 nongravid and 1 subgravid female;September: 1 male; October: 2 males and 1 gravid female with larvae. In a previous study carried outat this locality in 1993 (Moravec et al. 1995a, c), the only male of N. caudatus was collected from R.guatemalensis in October. These records suggest that males and gravid females occur more frequentlyin the autumn months (September, October), although it is highly probable that there areno pronounced seasonal maturation cycles in N. caudatus.Fig. 9. Monthly changes in mean numbers of specimens of three stages of Rhabdochona kidderi Pearse per fish.A = larvae and females without eggs; B = males; C = females with immature eggs; D = females with mature eggs.133

Tab. 8. Monthly survey of Rhamdia guatemalensis (Günther) from the cenote Ixin-há for infection with Rhabdochonakidderi Pearsemonth no. of fish examined no. of fish infected prevalence (%) intensity mean (range)June 20 14 70 7.6 (1–26)July 1 7 1 7 100 7.7 (1–38)August 14 12 86 12.6 (2–32)September 14 10 71 6.6 (2–17)October 15 13 87 7.7 (1–26)November 2 1 50 4.0 (4)Rhabdochona kidderi Pearse, 1936In Yucatán, this nematode (its nominotypical subspecies) mainly occurs in the intestine of R. guatemalensisand less often that of the blind cave fish, Ogilbia pearsei, in cenotes and caves (Moravec1998); from the cenote Ixin-há it was reported by Moravec et al. (1995a). R. kidderi occurs inMexico and the southern USA (Texas).Occurrence of Rhabdochona kidderi in Rhamdia guatemalensisPrevalence and intensity of infection: Out of 82 R. guatemalensis examined, 67 (prevalence 82%)were infected with R. kidderi, and the intensity of infection was 1–38 (mean 8) nematodes per fish.The number of catfish examined and their infection with R. kidderi is given in Tab. 8. Infectionby this nematode was associated with the body size (age) of the host (Tab. 2). The smallest specimenharbouring R. kidderi measured 12 cm and the largest 27 cm. The nematodes occurred in allsize-groups of catfish examined. The smallest size-groups were not examined. Tab. 2 shows that thepercentage of fish infected decreased with increase of body length of the host. The mean intensityof infection was highest in the smallest (15 cm and less) fish, and somewhat lower in the middle-sizedfish than in the the largest fish (20 cm and more).Seasonal changes in prevalence and mean intensity of infectionA survey of the monthly prevalence, intensity and mean intensity of R. kidderi infection in catfishis shown in Tab. 8. R. kidderi was present in catfish during the whole period. Prevalence wasusually high, at its maximum (100%) in July, decreased gradually in August and September (71%),increased in October (87%), and decreased again in November (Fig. 7). The mean intensity washighest (12.6) in August and lowest in September (6.6) (ignoring the data for November, when onlytwo fish were examined).Seasonal changes in maturation of Rhabdochona kidderiMonthly changes in the occurrence and the state of maturity of R. kidderi in R. guatemalensis areshown in Figs 8 and 9. The larvae and non-gravid females of this parasite occurred from June toOctober, with the highest percentages (22% and 23%) in July and September (Fig. 8). Malesoccurred from June to November, and made up a considerable part (31–51%) of all samples. Femaleswith immature eggs occurred from June to October; their proportion decreased from 31% in June to8% in August, increased slightly (14%) in September, and decreased markedly (2%) in October.Gravid females with mature eggs occurred from June to November; the proportion in samplesmarkedly increased in August and this trend continued until November (Fig. 8).The numbers of individual developmental stages of R. kidderi in R. guatemalensis each monthare shown in Fig. 9. The numbers of larvae and juvenile females per fish were approximately thesame (1.40–1.76) from June to September, but suddenly decreased (0.53) in October. The maximum134

number (5.57) of males occurred in August and the minimum (1.00) in November. Numbers offemales with immature eggs continuously decreased from 2.20 in June to 0.13 in October. Themaximum number (3.86) of females with mature eggs was found in August, the minimum (1.00) inNovember.Examination of invertebratesAn examination of aquatic invertebrates (see Material and Methods) did not reveal any larvalstages of the helminths parasitizing the fish, although the cystophorofurcocercous cercariae recordedin one of twelve aquatic snails, Pyrgophorus coronatus (Pfeiffer, 1839), examined on 13 July1994, probably belonged to G. tropica. The encysted metacercariae found in three out of the fiveunidentified dragon-fly nymphs (Odonata), examined on the same day (intensity 1–2 metacercariae),were identified as Loxogenes sp. (Trematoda: Lecithodendriidae), which as adults were foundin the intestine of the frog Rana brownorum Sanders, 1973 in the same locality (unpublished).The metacercariae were in round, thin-walled (3 µm) light-coloured cysts 30–43 µm in diameter.One specimen (Fig. 10), which was removed from a cyst and fixed in 4% formaldehyde, was oval, 490µm long and 225 µm wide. Body was densely covered by small tegmental spines visible at itsanterior part. The oral sucker was subterminal, size 55×60 µm, the ventral sucker situated in themiddle of body was slightly smaller, measuring 40×45 µm. The prepharynx was absent, the roundpharynx was 25 µm in diameter. The narrow oesophagus was 75 µm long. Caeca were short, extendingposteriorly to the level of the posterior end of the ventral sucker. The voluminous excretorypore was V-shaped, 200 µm long, and its anterior branches reached nearly to the level of the ventralsucker.DISCUSSIONAlthough the principal factor responsible for the seasonal maturation of fish helminths is watertemperature, this may operate via other factors, such as seasonal changes in the host’s physiology,availability of infective larvae (i. e. in the range of intermediate and paratenic hosts and cyclicalchanges in their abundance), and food preference and behaviour of the fish (Moravec 1998). It isFig. 10. Metacercaria of Loxogenes sp. from unidentified dragon-fly nymphs (Odonata) from the cenote Ixin-há.135

clear, therefore, that the seasonality in the occurrence and maturation of these parasites will be, inaddition to other factors, highly affected by their life-cycle patterns.The life cycle of the trematode Genarchella tropica in not known, but is likely to be similar tothat of the related congeneric species, G. astyanacis (Watson, 1976), a parasite of the characidAstyanax fasciatus (Cuvier, 1819) in Mexico and Nicaragua (Scholz et al. 1995a). According toDitrich et al. (1997) and Scholz et al. (2000), the first intermediate host of G. astyanacis in Yucatánis the aquatic snail Pyrgophorus coronatus, in which a cystophorofurcocercous cercaria develops,and the experimentally established second intermediate host is the copepod Mesocyclops chaciFiers, Reid, Iliffe et Suárez-Morales, 1996. The cystophorofurcocercous cercariae recorded duringthis study from P. coronatus (see p. 135) probably belonged to G. tropica, because no definitivehosts of other congeneric trematode species occur in this locality; apparently, various copepodsserve as the second intermediate host and a source of infection by this trematode for catfish.The present data suggest that new infections of G. tropica are acquired by catfish throughoutthe year, possibly because copepods are available all year. Although oviposition may occur all yearround, it mainly occurs in June and July. The quantitative differences may be associated withpossible seasonal changes in the abundance of the copepod intermediate hosts. The increase ininfection rate with increase in body size of the catfish may be associated with the amount of foodeaten (and consequently the numbers of infected copepods eaten) by fish, but it cannot be excludedthat small fish (including young catfish ) may serve as paratenic or paradefinitive hosts of G.tropica and be an addititional source of infection; this is suggested by the finding of a juvenile G.tropica specimen in Gambusia yucatana Regan, 1914 in the cenote Noc-ac in Yucatán by Scholzet al. (1995a).The first intermediate host of the trematode Stunkardiella minima is unknown, but it is possiblethat the oculopleurolophocercous cercariae in the snail Pyrgophorus coronatus at this locality(Ixin-há) and identified as Oligogonotylus manteri Watson, 1976 by Ditrich et al. (1997) and Scholzet al. (2000), are in fact cercariae of S. minima (T. Scholz pers. comm.). The definitive hosts of O.manteri are cichlids, which are not known to occur in the cenote Ixin-há. Metacercariae of S.minima have been reported from fish (mainly Rhamdia guatemalensis, rarely in Gambusia yucatana),which serve as the second intermediate hosts (Scholz et al. 1995b, Scholz & Aguirre-Macedo2000). The definitive host (R. guatemalensis) acquires the infection by cannibalism or predation onother fish species.The present data indicate that Stunkardiella minima metacercariae probably occur in catfish atthis locality throughout the year. Consequently, new infections are acquired by catfish as a definitivehost also all the year round, even though only nongravid or very young gravid specimens ofS. minima were recorded from June to August (this is probably due to the low numbers of fishsampled and the rapid development of the trematode in the definitive host at high water temperatures).It can be assumed that S. minima lays eggs in all months, but the highest prevalence and arelatively high mean intensity of gravid specimens were recorded in June. It is interesting that themean intensity of S. minima metacercariae was also highest in June. The frequency distribution andaggregation of infections by metacercariae indicate a considerable overdispersion of infections byyoung and adult trematodes.The life cycle of the cestode Proteocephalus brooksi is unknown, but it is assumed to be similarto that of other Proteocephalus spp., where various copepods serve as obligate intermediate hostsand the source of infection for the definitive host (Scholz 1999). Although fish paratenic hosts havebeen demonstrated for some species of this genus parasitizing piscivorous fishes, in the case of P.brooksi it is not clear whether R. guatemalensis harbouring encapsulated larvae of P. brooksi actsas a paratenic host or an obligate second intermediate host. However, even if there is only one136

intermediate host, copepod, catfish undoubtedly play an important role as a source of P. brooksiinfection for the definitive host, which it acquires by cannibalism.The present study indicates that there are large numbers of encapsulated larvae of P. brooksi incatfish throughout the year and are thus available for the definitive host. Consequently, it can beassumed that new infections are acquired by Rhamdia guatemalensis all the year round, but few ofthe acquired cestode larvae can establish and mature in the definitive host’s intestine, a phenomenonwell-known for other species of this genus (Scholz 1999). Even though gravid specimens of P.brooksi were only found in June and August, evidently as a result of a low rate of infection, it ishighly probable that the oviposition occurs throughout the year, in contrast to the majority ofcongeneric palaearctic species, which have pronounced seasonal maturation cycles and whereoviposition is restricted to 1–2 months in a year (Scholz 1999). This difference probably results fromthe source of infection (fish harbouring encapsulated cestode larvae are available throughout theyear vs. seasonal occurrence of infected copepods with a life of a few months) and relatively highwater temperatures. The decreasing prevalence and mean intensity of encapsulated P. brooksilarvae with increase in body length of R. guatemalensis reflects the fact that these fish acquire theinfection by feeding on copepod intermediate hosts.The life cycle of the nematode Neophilometroides caudatus is unknown, but other philometridsare known to utilize copepods as obligate intermediate hosts. Therefore, it is likely that the life cycleof N. caudatus also involves a copepod intermediate host, but the existence of a fish paratenic host(probably young catfish) as in some other philometrids parasitizing piscivorous fish (e. g., Philometraobturans (Prenant, 1886) parasitic in Esox lucius Linnaeus, 1758 in Europe – see Moravec &Dyková 1978) can be expected. The presence of a fish paratenic host of N. caudatus is suggestedby the fact that this parasite was recorded only in large R. guatemalensis.Even though males and the gravid female with larvae were recorded from catfish only in Septemberand October (apparently due to a low infection rate), it is highly probable that there is nopronounced seasonal maturation cycle in N. caudatus and that both acquiring new infections bythe definitive host and the production of larvae by gravid females take place throughout the year.As in the foregoing species, this may be associated with a relatively constant and high watertemperature, and a fish paratenic host that enables new infections to be acquired throughout theyear. All Philometra spp. from European cyprinids, where the only source of infection are copepods,have markedly pronounced seasonal maturation cycles with a short-term production of larvaemostly in late spring, but the principal source of Philometra obturans infection of pike, Esoxlucius, is fish paratenic hosts and, consequently, the production of larvae of this parasite occursthroughout the year (Moravec 1994).Intermediate hosts of various Rhabdochona spp. are larvae of aquatic insects (Ephemeroptera,Plecoptera, Trichoptera). The life cycle of R. kidderi texensis Moravec et Huffman, 1988 wasrecently studied by Moravec & Huffman (2001) who found the nymphs of the mayfly Tricorythodescurvatus Allen, 1977 to be the natural intermediate host of this parasite in Texas, USA, andalso successfully infected a European mayfly species. Seasonal cycles in the occurrence and maturationof R. kidderi in Cichlasoma nigrofasciatum (Günther, 1867) in the Amacuzac River incentral Mexico was studied by Caspeta-Mandujano et al. (2000). Although some unidentifiedephemeropteran and other aquatic insect larvae were examined from Ixin-há during this study (seeMaterials and Methods), no Rhabdochona larvae were recorded from them.The present data suggest that Rhabdochona kidderi occurs in Rhamdia guatemalensis in thecenote Ixin-há throughout the year, with the highest prevalence in July. Similar to what Caspeta-Mandujano et al. (2000) found for this species in central Mexico, both juveniles and gravid femaleswith mature eggs were recorded nearly all months, indicating that both new infections and ovipo-137

sition occurred throughtout the year. The main reason for this may be the annual presence ofsuitable ephemeropteran intermediate hosts in the locality and a rapid development of the nematodeboth in the intermediate and the definitive host. Quantitative differences in the monthlysamples of R. kidderi may be due to seasonal changes in populations of the mayfly intermediatehosts.Some European species of Rhabdochona (R. hellichi (Šrámek, 1901), R. phoxini Moravec, 1968)exhibit highly pronounced maturation cycles, where the oviposition is restricted to a short period inlate spring and summer (Moravec 1977, Moravec & Scholz 1995), whereas oviposition in R. denudata(Dujardin, 1843) occurs all the year round (Moravec 1989); the main reason for these differencesis the seasonal changes in the availability of infective larvae of these nematodes due to theseasonal occurrence of mayfly intermediate hosts in addition to the direct effect of temperature(Moravec 1994). An indistinct seasonal maturation cycle is found in R. zacconis Yamaguti, 1935from the subtropical region of Japan (Moravec et al. 1998).It is evident from the results of this study that none of the five helminth species from R. guatemalensisin the cenote Ixin-há exhibit a clear-cut seasonal maturation cycle with reproductionrestricted to a certain season. In this they differ from the majority of adult helminths parasitizingfreshwater fish in the temperate zone. Similar results were obtained by Caspeta-Mandujano et al.(2000) studying the seasonality of the nematode Rhabdochona kidderi in Cichlasoma nigrofasciatumin another tropical region of Mexico; in the Amacuzac River in the State of Morelos.It appears that in the tropics the main factor enabling fish helminths to reproduce throughout theyear is a relatively high water temperature all the year round, which enables the parasites to rapidlydevelop, and the quickly changing generations of invertebrate intermediate hosts. Some quantitativeseasonal differences in the populations of fish helminths are, apparently, associated with theecology and ethology of their intermediate, paratenic and definitive hosts.The association of infection rates with body size of host seems to depend on the way of acquiringinfection by the host. In those species where infection is acquired by feeding on small invertebrateintermediate hosts, i.e. Genarchella tropica, Rhabdochona kidderi and Proteocephalusbrooksi larvae, the rate of infection decreases with increasing body length of the host. In contrast,those species acquiring infection by feeding on fish intermediate or paratenic hosts, e. g. P. brooksi,the rate of infection increases with increasing body length of the host. This relationship is notapparent in Stunkardiella minima and Neophilometroides caudatus, where other ecological factorsmight operate.A c k n o w l e d g e m e n t sWe wish to thank fellow-workers at the Laboratory of Parasitology and Histopathology of the CINVESTAV-IPNMérida, namely R. Simá-Alvarez, J. Güemez Ricalde and G. Arjona-Torres for their assistance in collecting fish,and I. Husáková from the Institute of Parasitology, ASCR, in eské Budjovice for her technical help with thepreparation of the manuscript. This study was supported by grant No. P099 from the Comisión Nacional paraConocimiento y Uso de la Biodiversidad (CONABIO), Mexico and by grant No. A6022901 from the Grant Agencyof the Academy of Sciences of the Czech Republic.REFERENCESCASPETA-MANDUJANO J. M., MORAVEC F., DELGADO-YOSHINO M. A. & SALGADO-MALDONADO G. 2000: Seasonalvariations in the occurrence and maturation of the nematode Rhabdochona kidderi in Cichlasoma nigrofasciatumof the Amacuzac River, Mexico. Helminthologia 37: 29–33.CHUBB J. C. 1979: Seasonal occurrence of helminths in freshwater fishes. Part II. Trematoda. Adv. Parasitol. 17:141–313.138

CHUBB J. C. 1982: Seasonal occurrence of helminths in freshwater fishes. Part IV. Adult Cestoda and Nematoda.Adv. Parasitol. 20: 1–292.DITRICH O., SCHOLZ T., AGUIRRE-MACEDO L. & VARGAS-VÁZQUEZ J. 1997: Larval stages of trematodes fromfreshwater molluscs of the Yucatan Peninsula, Mexico. Folia Parasitol. 44: 109–127.HALL F. G. 1977: Cenotes y aguadas. Pp.: 67–80. In: Enciclopedía Yucatanense. Tomo I. Gobierno del Estado deYucatán.HAMANN M. I. 1999: Population biology of Spirocamallanus inopinatus (Travassos, Artigas et Pereira, 1928)(Nematoda: Camallanidae) in Serrasalmus spiropleura Kner, 1860 (Pisces: Characidae) from Corrientes,Argentina. Res. Rev. Parasitol. 59: 1–6.KENNEDY C. R. 1970: The population biology of helminths of British freshwater fish. In: TAYLOR A. E. R (ed.):Symp. Br. Soc. Parasitol. 8: 145–159.LEE J.C. 1996: The amphibians and reptiles of the Yucatán Peninsula. Ithaca: Cornell University Press, 500 pp.MORAVEC F. 1977: Life history of the nematode Rhabdochona phoxini Moravec, 1968 in the Rokytka Brook,Czechoslovakia. Folia Parasitol. 24: 97–105.MORAVEC F. 1989: Observations on the occurrence and maturation of the nematode Rhabdochona denudata(Dujardin, 1845) in chub, Leuciscus cephalus (L.), of the Rokytná River, Czechoslovakia. Parassitologia 31:25–35.MORAVEC F. 1994: Parasitic nematodes of freshwater fishes of Europe. Praha and Dordrecht, Boston, London:Academia and Kluwer Acad. Publishers, 473 pp.MORAVEC F. 1998: Nematodes of freshwater fishes of the Neotropical Region. Praha: Academia, 464 pp.MORAVEC F. & DYKOVÁ I. 1978: On the biology of the nematode Philometra obturans (Prenant, 1886) in thefishpond system of Mácha Lake, Czechoslovakia. Folia Parasitol. 25: 231–240.MORAVEC F. & HUFFMAN D. G. 2001: Observations on the biology of Rhabdochona kidderi texensis, a parasite ofNorth American cichlids. J. Helminthol. 75: 197–203.MORAVEC F., NAGASAWA K. & URUSHIBARA Y. 1998: Observations on the seasonal maturation of the nematodeRhabdochona zacconis in Japanese dace, Tribolodon hakonensis, of the Okitsu River, Japan. Acta Soc. Zool.Bohem. 62: 45–60.MORAVEC F., SALGADO-MALDONADO G. & AGUILAR-AGUILAR R. 2002: Neophilometroides n. gen. (Nematoda:Philometridae) for Philometroides caudatus Moravec et al., 1995, with erection of Neophilometrinae n.subfam. J. Parasitol. (in press).MORAVEC F. & SCHOLZ T. 1995: Life history of the nematode Rhabdochona hellichi, a parasite of the barbel in theJihlava River, Czech Republic. J. Helminthol. 69: 59–64.MORAVEC F., SCHOLZ T. & VIVAS-RODRÍGUEZ C. 1995c: Philometroides caudata sp. n. (Nematoda: Philometridae)from Rhamdia guatemalensis (Pisces) in Yucatan, Mexico. Folia Parasitol. 42: 293–298.MORAVEC F., VIVAS-RODRÍGUEZ C., SCHOLZ T., VARGAS-VÁZQUEZ J., MENDOZA-FRANCO E. & GONZÁLEZ-SOLÍS D.1995a: Nematodes parasitic in fishes of cenotes (= sinkholes) of the Peninsula of Yucatan, Mexico. Part 1.Adults. Folia Parasitol. 42: 115–129.MORAVEC F., VIVAS-RODRÍGUEZ C., SCHOLZ T., VARGAS-VÁZQUEZ J., MENDOZA-FRANCO E., SCHMITTER-SOTO J. J. &GONZÁLEZ-SOLÍS D. 1995b: Nematodes parasitic in fishes of cenotes (= sinkholes) of the Peninsiula of Yucatan,Mexico. Part 2. Larvae. Folia Parasitol. 42: 199–210.PÉREZ-PONCE DE LEÓN G., GARCÍA-PRIETO L., OSORIO-SARABIA D. & LEÓN-RGAGNON V. 1996: Listados faunísticode México VI. Helmintos parásitos de peces de aguas continentales de México. México: Universidad NacionalAutónoma de México, Instituto de Biología, 100 pp.SCHOLZ T. 1999: Life cycles of species of Proteocephalus, parasites of fishes in the Palearctic Region: A review. J.Helminthol. 73: 1–19.SCHOLZ T. & AGUIRRE-MACEDO M. L. 2000: Metacercariae of trematodes parasitizing freshwater fish in Mexico:A reappraisal and methods of study. Pp.: 101–115. In: SALGADO-MALDONADO G., GARCÍA ALDRETE A. N. &VIDAL-MARTÍNEZ V. M. (eds.): Metazoan parasites in the neotropics: A systematic and ecological perspective.Mexico: Instituto de Biología, Universidad Nacional Autónoma de México, 310 pp.SCHOLZ T., AGUIRRE-MACEDO M. L., SABASFLORES DÍAZ DE LEÓN A. T. & DITRICH O. 2000: Larval stages oftrematodes in Mexican freshwater molluscs: A review of present state and methodology for future research.Pp.: 77–100. In: SALGADO-MALDONADO G., GARCÍA ALDRETE A. N.& VIDAL-MARTÍNEZ V. M. (eds.): Metazoanparasites in the neotropics: A systematic and ecological perspective. Mexico: Instituto de Biología, UniversidadNacional Autónoma de México, 310 pp.SCHOLZ T., VARGAS-VÁZQUEZ J., MORAVEC F., VIVAS-RODRÍGUEZ C. & MENDOZA-FRANCO E. 1995a: Cenotes (sinkholes)of the Yucatan Peninsula, Mexico, as a habitat of adult trematodes of fish. Folia Parasitol. 42: 37–47.139

SCHOLZ T., VARGAS-VÁZQUEZ J., MORAVEC F., VIVAS-RODRÍGUEZ C. & MENDOZA-FRANCO E. 1995b: Metacercariaeof trematodes of fishes from cenotes (= sinkholes) of the Yucatan Peninsula, Mexico. Folia Parasitol. 42:173–192.SCHOLZ T., VARGAS-VÁZQUEZ J., MORAVEC F., VIVAS-RODRÍGUEZ C. & MENDOZA-FRANCO E. 1996: Cestoda andAcanthocephala of fishes from cenotes (= sinkholes) of Yucatan, Mexico. Folia Parasitol. 43: 141–152.SCHOLZ T., VARGAS-VÁZQUEZ J. & SALGADO-MALDONADO G. 1995c: Revision of Genarchella species (Digenea:Derogenidae) parasitizing freshwater fishes in Mexico and Central America. J. Natur. Histor. 29: 1403–1417.140

Acta Soc. Zool. Bohem. 66: 141–150, 2002ISSN 1211-376XNew Tertiary dragonflies from Lower Oligocene of the eské stedohoíMts and Lower Miocene of the Most Basin in the Czech Republic(Odonata: Anisoptera)Jakub PROKOP 1, 2) & André NEL 3)1)Department of Zoology, Charles University, Vininá 7, CZ–128 44, Praha 2, Czech Republic;e-mail: jprokop@natur.cuni.cz2)Department of Palaeontology, Charles University, Albertov 6, CZ–128 43 Praha 2, Czech Republic3)Laboratoire d’Entomologie et CNRS UMR 8569, Museum national d’Histoire naturelle, 45 rue Buffon,F–75005 Paris, France; e-mail: anel@mnhn.frReceived January 30, 2002; accepted March 23, 2002Published June 28, 2002Abstract. Two new representatives of the clade Aeshnoptera are described from the Lower Oligocene andLower Miocene of northern Bohemia (Czech Republic), i. e. Kvacekia infuscata gen. n. et sp. n. (Aeshnidae)and Gomphaeschna miocenica sp. n. (Gomphaeschnidae). Kvacekia gen. n. seems to be closely related tothe Cenozoic genus Oligaeschna Piton et Théobald, 1939 and the recent genus Oplonaeschna Selys, 1883.Gomphaeschna miocenica sp. n. wing venation has particular wing coloration and distinctly differentcharacters from all previously described species of the genus. A holarctic distribution in fossil history isproposed for both Oplonaeschninae and Gomphaeschninae.Taxonomy, fossil, description, Insecta, Anisoptera, Odonata, Aeshnoptera, Gomphaeschnidae,Aeshnidae, Kvacekia infuscata gen. n. et sp. n., Gomphaeschna miocenica sp. n., Tertiary, LowerOligocene, Lower Miocene, Central Europe, Palaearctic regionINTRODUCTIONTertiary fossil dragonflies have been recorded from several sites such as Pochlovice near Kynšperk,Sokolov, Jehliná near Sokolov belonging to Cypris Shale and from Bílina mine of Most Formationin northwestern Bohemia (Bachmayer 1952, Heer 1849, Handlirsch 1907, Kukalová 1962, íha 1977,1979, Prokop & Nel 2000). Up till now all these recorded fossils, both larvae and adults, belong toLibellulidae and Aeshnidae (Prokop 2002).Cenozoic insect fauna of northwestern Bohemia is preserved in fluvio-lacustrine deposits ofthe Krušné hory piedmont basins and the eské stedohoí Mts. About 16 localities representingseveral different palaleoenvironments dated from Upper Eocene to Lower Miocene are investigated.The relatively diverse insect fauna from Lower Oligocene (Ruppelian/Chattian) of Kundraticenear Litomice is preserved in several interbeds of the brownish diatomite of Ústí Formation (Fig.1b). The site is well known because of studied palaeobotanical and palaeoichthyological records(Kvaek & Walther 1998, Obrhelová 1979). Another famous site is the Bílina mine (the formerMaxim Gorkij mine) situated in the Most Basin near the town of Bílina (Fig. 1b). The stratigraficalattribution is to the Most Formation of the Lower Miocene (Eggenburgian/Ottnangian) and insectsare preserved in three fossiliferous horizons (Clayey Superseam Horizon, Delta Sandy Horizon,Lake Clayey Horizon) (Prokop 2002). The site has been observed from other paleoscientificaspects as palaeobotany, palynology and sedimentology (Bžek et al. 1992, Dašková 2000, Kvaek1998, Rajchl & Uliný 1999, Sakala 2000).141

In the following study we use the wing venation nomenclature of Riek & Kukalová-Peck (1984),amended by Kukalová-Peck (1991), Nel et al. (1993), Bechly (1996) and Bechly et al. (2001). Wefollow the phylogenetic classification of Anisoptera proposed by Bechly et al. (1996, 2002).SYSTEMATIC PALAEONTOLOGYAeshnoptera Bechly, 1996Family Aeshnidae Leach, 1815Genus Kvacekia gen. n.TYPE SPECIES. Kvacekia infuscata sp. n.DIAGNOSIS. The new genus very similar and probably closely related to recent genus OplonaeschnaSelys, 1883 and fossil genus Oligaeschna Piton et Théobald, 1939. It differs from these generaby the following combination of characters (fore wing structures): (1) pterostigma elongate, covering5–6 cells; (2) cubito-anal area very broad, with more than 9 rows of cells between CuA andposterior wing margin (possible autapomorphy, but character unknown in some Oligaeschnaspecies); (3) 5 rows of cells in area between IR2 and RP2; (4) hypertriangle with 4 or more crossveins;(5) wing dark infuscate; (6) about 18–20 postnodal cross-veins (14 to 16 in the Oplonaeschnaand Oligaeschna species, but character state unknown in Oligaeschna lapidaria). Characters(2) and (5) are probably autapomorphies of Kvacekia gen. n., but no relative autapomorphy ofOligaeschna is known. Only the discovery of better-preserved specimens of both taxa will permitto solve the problem of the possible paraphyly of Oligaeschna relatively to Kvacekia gen. n.ETYMOLOGY. Kvacekia gen. n. (feminine in gender), named in honour of Prof. Zlatko Kvaek,palaeobotanist from Charles University in Prague.Kvacekia infuscata sp. n.(Figs 2, 3)DIAGNOSIS. That of the genus.DESCRIPTION. A nearly complete fore wing, broken in nodal region and with part basal of arculusmissing; whole wing surface dark fuscous with some lighter zones (see Fig. 2); fore wing, about52 mm long (assumed from fragment) and 12 mm wide; distance from estimate base to nodus, about25 mm; distance from nodus to wing apex, about 27 mm; nodus nearly midway between base andapex; distance from nodus to pterostigma, 12.4 mm; distance from pterostigma to apex, 5.6 mm;pterostigma rather long, 5.3 mm long and 0.9 mm wide, covering approximately 5–6 cells; pterostigmalbrace slightly obliquely aligned with proximal side of pterostigma; 19 visible antenodal crossveinsof first row between C and ScP not aligned with the 16 visible corresponding antenodalcross-veins of second row between ScP and RA; 18 visible postnodal cross-veins, not well alignedwith 19 visible subpostnodal cross-veins, maybe 1–2 missing postnodal cross-veins close tonodus; more than 12 secondary antenodal cross-veins visible between Ax2 and nodus; more than4 antenodal cross-veins of second row between Ax2 and Ax1; Ax1 being 6.2 mm and Ax2 12.9 mmfrom wing base; hypertriangle crossed by 4–5 cross-veins; arculus, anal area, median and submedianspace, and subdiscoidal triangle not preserved; discoidal triangle elongate and divided into7 smaller cells, its costal side being 9.2 mm long, distal side 8.3 mm long and proximal side 3.1 mmlong; width of postdiscoidal area just behind discoidal triangle, 4 mm, probable width along posteriorwing margin, 11.4 mm; 3 rows of cells in postdiscoidal area just distal of discoidal triangle; ashort convex supplementary sector (trigonal planate) in postdiscoidal area, beginning one cell142

efore distal angle of discoidal triangle, and aligned with concave Mspl; Mspl well defined, nearlystrait in its basal half but slightly curved in distal part; 2 rows of cells between Mspl and MP and5 rows between Mspl and MA; 2 rows of cells and a zigzagged supplementary vein in distal part ofarea between MA and RP3/4; bulge in distal part of MA (“aeshnid bulla”) weak; 2 preserved Bqcross-veins; oblique vein ’O’ not preserved; Rspl well defined and slightly anteriorly curved in itsdistal part; area between Rspl and IR2 very wide, with about 5 rows of cells in its widest part; spacebetween IR2 and Rspl basally divided by oblique intercalary veinlets; IR2 smoothly curved distallybut not forked; a long and rather straight intercalary vein, close and well parallel to RP2, proximallynot branching on IR2 but vanishing in area between RP2 and IR2; RP2 strongly curvedposteriorly opposite proximal side of pterostigma; 5 rows of cells between RP2 and IR2 in widestpart; IR1 present but zigzagged, 6.8 mm long, beginning just below proximal side of pterostigma;one row of cells between MP and CuAa; CuAa with more than 8 posterior branches directedtowards posterior wing margin; cubito-anal area very broad.TYPE MATERIAL. Holotype: specimen P1193 (National Museum coll., Praha, Czech Republic), medio-distal part offore wing (imprint) with venation well preserved, only posterior part of wing preserved in counter imprint.TYPE LOCALITY. Kundratice near Litomice, Czech Republic.TYPE STRATA. Lower Oligocene (Ruppelian/Chattian), Ústí Formation (Stedohoí Complex).ETYMOLOGY. Named after the dark infuscate wing of the type specimen.DISCUSSION. Kvacekia gen. n. fits into the Aeshnidae because of the presence of several potentialautapomorphies (after Bechly 1996): “Rspl and Mspl distinctly curved, with more than 3 rows ofFig. 1. Geographical position of northwestern Bohemia within Europe (A), detailed map of the Most Basin andthe eské stedohoí volcanic areas (B), 1 – Bílina mine, 2 – Kundratice near Litomice.143

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cells between them and IR2 or MA”; “space between IR2 and Rspl basally divided by obliqueintercalary veinlets”;”more than 2 rows of cells in basal part of postdiscoidal area between level ofdistal angle of discoidal triangle and level of midfork”; “hypertriangles traversed by at least 3cross-veins in forewings”. As all these characters are convergently present in some other aeshnopteridgroups, it is difficult to consider them as very accurate characters. Nevertheless, theircombination is only present in Aeshnidae. Within this family, Kvacekia gen. n. shares with theOplonaeschninae Bechly, 1996 the unique autapomorphy of this subfamily, “IR2 unbranched”,that Bechly (1996, 2002) considered as a reversion. Note that some Aeshna spp. (among others, A.coerulea (Ström, 1783); A. septentrionalis Burmeister, 1839; A. ollivieri Nel, 1986) have a veryrudimentary division of IR2 into a strong posterior branch and a weak more or less zigzaggedanterior branch (Nel et al. 1994). Kvacekia gen. n. has no trace of such an anterior branch of IR2.Bechly (1996, 2002) included in this group the recent genus Oplonaeschna Selys, 1883 (plusmaybe Basiaeschna Selys, 1883). Kvacekia gen. n. differs from Basiaeschna and Oplonaeschnain the following characters: “presence of 4 cross-veins in fore wing hypertriangle, instead of 2 inBasiaeschna and 3 in Oplonaeschna’, “broader postdiscoidal area with 3 rows of cells just distalof discoidal triangle, instead of 2 in both Basiaeschna and Oplonaeschna”, “broader areas betweenMspl and MA and between Rspl and IR2, with 5 rows of cells, instead of 3–4 rows inOplonaeschna and 2–3 rows in Basiaeschna”. Furthermore, it differs from Oplonaeschna in thefollowing characters: “pterostigma distinctly longer covering 5–6 cells instead of 3 in Oplonaeschna”.The pterostigma of Basiaeschna is covering 4 cells. Nel et al. (1994) revised the Oligocenegenus Oligaeschna Piton and Théobald, 1939 (type species O. jungi Piton et Théobald, 1939) andconsidered it as closely related to Oplonaeschna (maybe its sister genus). They also transferred 4fossil species from Oplonaeschna into Oligaeschna, i. e. O. separata (Scudder, 1890), O. lapidaria(Cockerell et Counts, 1913), O. ashutasica (Martynov, 1929) and O. palaeocoerulea (Timon-David, 1946). Kvacekia gen. n. shares with Oligaeschna its long pterostigma (plesiomorphy), andits postdiscoidal area with 3 rows of cells just distal of discoidal triangle (synapomorphy) (Nel etal. 1994). It differs from Oligaeschna jungi Piton et Théobald, 1939 and O. separata in its distinctlybroader cubito-anal area, with about 9 rows of cells between CuA and posterior fore wing margin,instead of 5 in O. jungi and O. separata (Scudder 1890: Pl. 13, Fig. 15). Unfortunately, this structureis missing in the type specimen of O. palaeocoerulea. Nevertheless, this last species is characterizedby its very broad area between IR2 and RP2, with 9 rows of cells along posterior wing margin,instead of 5 rows in Kvacekia gen. n. and O. jungi (direct exam of type specimen, Timon-David,1946). Oligaeschna lapidaria is based on a hind wing. Thus, it is not possible to compare thewidth of its cubito-anal area to that of the fore wing of Kvacekia gen. n. Nevertheless, it differsfrom Kvacekia gen. n. in its pterostigma covering only 3 cells, instead of 5–6 in Kvacekia gen. n.and in its hyaline wing (Cockerell 1913). Oligaeschna ashutasica is based on a hind wing. Neverthelessits pterostigma is covering 4 cells instead of 5–6 in Kvacekia gen. n., and it has 3 rows ofcells between IR2 and RP2, instead of 5 in Kvacekia gen. n. (Martynov 1929: text and fig. 4).Nothing is known about the wing coloration of Oligaeschna jungi because no trace of colorationis preserved in all known specimens, and of O. ashutasica, as Martynov (1929) indicatednothing about this point. All other Oligaeschna species have hyaline wings, as many recentAeshnidae. Some recent Aeshnidae (among others, Subaeschna francesca Martin, 1909 and Agyrt-Figs 2–5. 2, 3 – Kvacekia infuscata gen. n. et sp. n., holotype specimen P1193 – photo and line drawing offorewing venation. Scale 5 mm. 4–5 – Gomphaeschna miocenica sp. n., holotype specimen ZD0184 – photo andline drawing of forewing venation. Scale 3 mm.145

acantha disrupta (Karsch, 1889) have a wing coloration very similar to that of Kvacekia gen. n.(Martin 1908).NOTE. Three fossil species Oplonaeschna vectensis Cockerell et Andrews, 1916, Oplonaeschnastaurophlebioides Henriksen, 1922 and Oplonaeschna metis (Heer, 1849) are based on ratherpoorly preserved specimens. Nel et al. (1994) considered them as Aeshnidae of uncertain position.Family Gomphaeschnidae Tillyard et Fraser, 1940 (sensu Bechly 1996 and Bechly et al. 2001)Gomphaeschna miocenica sp. n.(Figs 4, 5)DIAGNOSIS. This species differs from all other described Gomphaeschna in the following points: (1)wing not hyaline but with all veins underlined by dark coloration and central part of each cellhyaline; (2) undulated IR2; (3) area between IR2 and RP2 not narrowed near wing margin, with 3rows of cells; (4) only 3 rows of cells between RP1 and RP2 along wing margin; (5) pterostigmarelatively long, covering 2 cells.DESCRIPTION. Distal part of a forewing (wing rather narrow below nodus); with all veins underlinedby dark coloration and central part of each cell hyaline; probable length, about 40 mm; distancebetween nodus and apex, 19.0 mm, between nodus and pterostigma, 11.1 mm, between pterostigmaand apex, about 4.3 mm; pterostigma, 3.3 mm long and 1.1 mm wide, with its costal and posteriorsides slightly widened in middle; pterostigmal brace not elongate, straight and only slightly oblique;pterostigma covering 2 cells; 6 postnodal cross-veins not aligned with the correspondingpostsubnodal cross-veins; 5 antenodal cross-veins of first row preserved, not aligned with thoseof second row, last antenodal cross-vein of first row incomplete, i. e. with no corresponding crossveinof second row; 2 preserved cross-veins between RA and RP basal of subnodus, presence ofthe antesubnodal gap; 2 Bq cross-veins between IR2 and RP2; MP curved and reaching posteriorwing margin opposite nodus; 3 rows of cells in postdiscoidal area just basal of Mspl; Mspl slightlycurved, distally weakly zigzagged, with 1–2 row of cells between it and MA; RP3/4 and MA arebasally nearly straight but distally curved and nearly perpendicular to posterior wing margin; ashort zigzagged intercalary vein between RP3/4 and MA in their distal part, but not ending onposterior wing margin; RP2 aligned with subnodus; oblique cross-vein ‘O’ 1.1 mm distal of subnodus;IR2 nearly straight; RP2 with 2 weak but distinct undulations; IR2 and RP2 distally curvedand nearly perpendicular to posterior wing margin; area between IR2 and RP2 rather widened in itsdistal half, with 2–3 rows of cells between them; a nearly straight vein Rspl, slightly curved in itsdistal end, with one row of cells between it and IR2; area between RP1 and RP2 rather wide, with 3rows of cells between them, but incompletely preserved, with a rather short and zigzagged IR1.TYPE MATERIAL. Holotype: specimen ZD0184 (Bílina mine coll., Czech Republic), distal half of fore wing (imprint)with venation well preserved.TYPE LOCALITY. Bilina mine, Czech Republic.TYPE STRATA. Lower Miocene (Eggenburgian/Ottnangian), Most Formation, Clay Superseam Horizon.ETYMOLOGY. Named after the Miocene age of the fossil.DISCUSSION. The specimen is clearly attributable to the clade Aeshnoptera because of the presenceof a clear vein Rspl, undulation of RP3/4 and MA and the narrow area between RP1 and RP2 basalof pterostigma. It can be included in Gomphaeschnidae Tillyard et Fraser, 1940 (sensu Bechly 1996and Bechly et al. 2001) because it has one of its main synapomorphies, viz. “the most distal part ofthe antesubnodal area between RA and RP is free of antesubnodal cross-veins”, although this146

character is less pronounced in Oligoaeschna Selys, 1889 (in other Gomphaeschna Selys, 1871species and Gomphaeschna miocenica sp. n., the most distal cross-vein of this area is oppositethe base of IR2, but there is one more in a slightly distal position at least in Oligoaeschna pryeri(Martin, 1909) and Oligoaeschna modigliani (Selys, 1889).Within this family, the autapomorphy of the fossil subfamily Gomphaeschnaoidinae Bechly etal., 2001 that could be visible in G. miocenica sp. n. is absent in this taxon: its pterostigmal bracevein is not very oblique. The other autapomorphies of the Gomphaeschnaoidinae are unknown inG. miocenica sp. n.: “only a single secondary antenodal cross-vein between Ax1 and Ax2 alignedlike a primary antenodal bracket” and “in forewing, Ax2 shifted basally on level of basal angle ofdiscoidal triangle”. As G. miocenica sp. n. has a plesiomorphic state of character, it is not sufficientto exclude it from this subfamily. Also, the other subfamily “Gomphaeschninae” (sensu Bechly etal. 2001) has no known autapomorphy and could be paraphyletic. Nevertheless, G. miocenicasp. n. is very different from all Gomphaeschnaoidini in its pterostigmal brace not undulating, theabsence of any basally widened cell below pterostigma, its cross-veins between RP2 and RP1 allsimilar, not oblique. It also differs from the unique representative of the Sinojagorini Bechly et al.,2001 (Sinojagoria Bechly et al., 2001) in its less numerous postnodal cross-veins and in the basalpart of area between RP2 and RP1 (6 against 10–12). Lastly, it differs from the Lower CretaceousAnomalaeschna berndschusteri Bechly et al., 2001 in its distinctly longer pterostigma, covering 2cells instead of only one.Within “Gomphaeschninae”, the recent genus Linaeschna Martin, 1909 is very different fromG. miocenica sp. n. in its very long postnodal area with numerous cross-veins and its absence ofgap in the distal part of antesubnodal area (Martin 1909). Bechly et al. (2001) considered that thisgenus could belong to a different family of the clade Aeshnoptera. The remaining genera areGomphaeschna, Oligoaeschna, Alloaeschna Wighton et Wilson, 1986 and Cretalloaeschna Jarzembowskiet Nel, 1996.The Lower Cretaceous Cretalloaeschna differs from G. miocenica sp. n. in the following characters:(1) presence of one row of cells between RP3/4 and MA, (2) a straight IR2, (3) only 2 rowsof cells between IR2 and RP2, (4) 4–5 rows of cells between RP2 and RP1, instead of 3 in G.miocenica sp. n. (Bechly et al. 2001). These authors indicated that character (1) could represent asynapomorphy of Cretalloaeschna with Gomphaeschna, Gomphaeschnoides Carle et Wighton,1990 (in Gomphaeschnaoidini), and Alloaeschna, but some specimens attributed to Alloaeschnapaskapooensis Wighton et Wilson, 1986 have 2 rows of cells defining a long zigzagged veinbetween RP3/4 and MA. Thus, this character is at least not so important, if subject to intraspecificvariation.The Late Paleocene genus Alloaeschna (at least A. paskapooensis and maybe A. marklaeWighton et Wilson, 1986) shares the autapomorphies proposed by Bechly et al. (2001) for theGomphaeschnaoidini, i. e. “presence of a characteristic elongate distal paranal cell in hindwing,longer than wide, directly basal of the anal loop” (see Wighton & Wilson 1986: Figs 1–10). InOligoaeschna and Gomphaeschna, the corresponding cell is quadrate or transverse, wider thanlong; “pterostigmal brace vein somewhat undulating”; “basally widened cell bellow pterostigma,caused by a curvature of RP1 at pterostigmal brace”; “posterior branches of CuAa relativelyweakly defined in hind wing” (at least in some specimens of Alloaeschna paskapooensis). Thus,we propose to transfer Alloaeschna to this tribe, previously only known in Lower Cretaceous. Itwould be its most recent representative in Early Cenozoic. As noted above, the preserved correspondingcharacters of pterostigmal region are different in Gomphaeschna miocenica sp. n.It is extremely difficult to separate the two genera Gomphaeschna and Oligoaeschna on thesole basis of the wing venation characters. Madsen & Nel (1997) proposed to use the presence of147

only 1–2 Bq cross-veins in Gomphaeschna instead of 3 in Oligoaeschna. Bechly et al. (2001: 197)attributed the two Cretaceous species ?Gomphaeschna inferna Pritykina, 1977 and ?G. sibiricaBechly et al., 2001 to the genus Gomphaeschna on the basis of “a reduced wing venation withfewer rows of cells between the main veins, and a short pterostigma with only one or two cellsbeneath it.” G. miocenica sp. n. has 2 Bq cross-veins and a short pterostigma covering only 2 cells.Thus, we tentatively attribute it to Gomphaeschna rather than to Oligoaeschna.Gomphaeschna miocenica sp. n. differs from ?G. inferna in its undulated IR2 and area betweenIR2 and RP2 not narrowed near wing margin, with 3 rows of cells, instead of 2 in ?G. inferna. G.miocenica sp. n. differs from ?G. sibirica in the presence of only 3 rows of cells between RP1 andRP2 along wing margin, instead of 7. G. miocenica sp. n. differs from recent Gomphaeschnafurcillata (Say, 1839) in its longer pterostigma covering 2 cells, instead of one cell and a half, thepresence of 3 rows of cells between IR2 and RP2, instead of 1–2 rows and its IR2 distinctlyundulated. G. miocenica sp. n. also differs from the two Paleocene/Eocene Gomphaeschna paleocenicaMadsen et Nel, 1997 and ?Gomphaeschna danica Madsen et Nel, 1997 in its IR2 undulated,instead of being straight. Another important difference with all species of Gomphaeschna isthe particular wing coloration of G. miocenica sp. n. All recent and fossil Gomphaeschna havehyaline wings (maybe except ?G. sibirica only known after a fossil with no preserved coloration).PALAEOGEOGRAPHIC REMARKSRecent species of Oplonaeschna and Basiaeschna are North American. The fossil genus Oligaeschnais known from the Oligocene of France, USA and Siberia. This suggests a Holarctic distributionof the Oplonaeschninae but little is known about the Cenozoic record of Gondwanian Aeshnoptera.Recent species G. furcillata (Say, 1839) and G. antillope Hagen, 1874 are distributed in theNorth America (USA), fossil members are known from Lower Cretaceous (?G. inferna – Baissa inBuryat Republic, Sibieria and ?G. sibirica – Chita region, Siberia, Russia) and Upper Paleocene/Lower Eocene (G. paleocenica, ?G. danica – Moclay in Denmark). Other related genus Oligoaeschnais recently living in South-east Asia and fossil representatives are known from UpperEocene: O. anglica Cockerel et Andrews, 1916 – Gurnet Bay, Isle of Wight, UK; Lower Oligocene:O. needhami (Cockerell, 1913) – Florissant, Colorado, USA, O. oligocenica (Nel et Papazian, 1983)– Aix-en-Provence, France, O. conjuncta (Martynov, 1929) – Ashutas Mountains, Zaisan, Siberia;Lower Miocene: ‘Projagoria conjuncta Martynov, 1929’ (sensu Zeuner 1938) – “Mainzer Hydrobienkalks”,Germany, Oligoaschna sinica (Zhang, 1989) comb. nov. – Miocene, Shanwang, Shandong,China, and Upper Pliocene: Oligoaeschna sp. (Esaki & Asahina 1957, Nel et al. 1994) –Nagasaki Prefecture, Japan. The fossil genus Alloaeschna is known from Paleocene (A. paskapooensis,A. marklae Wington et Wilson, 1986, A. quadrata Wighton et Wilson, 1986 – Blackfaldsin Alberta, Canada). All other representatives of the Gomphaeschnaoidinae are from the LowerCretaceous of China, Mongolia, UK and Brazil. From these palaeogeographical data we can assumea very wide, maybe almost cosmopolitan distribution of the family during the Lower Cretaceous,a wide Holarctic distribution of the subfamily Gomphaeschninae during the Tertiary and amore marginal occurrence of the taxa present today.A c k n o w l e d g e m e n t sThe authors are grateful to Zdenk Dvoák (Doly Bílina) and Kamil Zagoršek (National Museum Prague) for theloan of the material. The name of Kvacekia gen. n. is dedicated to 65 birthday of Prof. Zlatko Kvaek, leader ofCzech palaeobotany at Charles University. The research was supported by grants of the Ministry of Schools J13/98-113100004, GAUK 197/2000 and GAR 205/01/0639.148

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Acta Soc. Zool. Bohem. 66: 151–160, 2002ISSN 1211-376XMonogyny in Leptothorax slavonicus (Hymenoptera: Formicidae)Klára TICHÁ 1) & Pavel ŠTYS 2)1)Muzeum Vysoiny, Masarykovo námstí 55, CZ–58601 Jihlava, Czech Republic2)Department of Zoology, Charles University, Vininá 7, CZ–12844 Praha 2, Czech RepublicReceived August 15, 2001; accepted October 6, 2001Published June 28, 2002Abstract. Monogyny in Leptothorax slavonicus Seifert, 1995 (Hymenoptera: Formicidae: Myrmicinae)was studied under field and laboratory conditions. None of 238 colonies from the Czech Republic andSlovakia were polygynous. Experiments based on mixing two monogynous colonies and offering twoqueens to an orphaned colony showed that this species is capable of forming genetically rather heterogeneousbut never polygynous colonies. Fusion of unrelated colonies and the adoption of alien queens occurred, butthe workers always regulated number of queens to one. The workers can recognise their nestmates inexperimentally mixed colonies.Ethology, monogyny, fusion of colonies, recognition of nestmates, Hymenoptera, Formicidae,Leptothorax slavonicus, Central EuropeINTRODUCTIONThis paper presents the results of an experimental study of monogyny in a recently establishedleptothoracine ant species Leptothorax (Myrafant) slavonicus Seifert, 1995 (Myrmicinae).The number of queens (see Terminology) present in a colony of eusocial insects markedlyaffects its genetic composition. About 50% of ant species form monogynous colonies (one queen)while the others may form polygynous colonies (several queens) (Buschinger 1974). There areobligatorily monogynous species, e. g. Lasius (Lasius) niger (Linnaeus, 1758), Camponotus (Camponotus)ligniperda (Latreille, 1802), Leptothorax (Myrafant) unifasciatus (Latreille, 1798); facultativelypolygynous species which form both monogynous and polygynous colonies, e. g. Leptothorax(Leptothorax) acervorum (Fabricius, 1793), Leptothorax (Leptothorax) muscorum (Nylander,1846), and obligatorily polygynous species, e. g. Formica (Formica) polyctena Förster,1850 (all cf. Buschinger 1967, 1968, 1974).Several unusual cases have been described in Leptothorax Mayr, 1855. The functionally monogynouscolony contains several inseminated females only one of which is the queen and laysfertilised eggs in a facultatively polygynous species Leptothorax (Leptothorax) gredleri Mayr,1855 (cf. Buschinger 1967, 1968, 1974, Heinze & Buschinger 1988) and some North Americanspecies of Leptothorax s. str. (cf. Heinze & Buschinger 1986, 1988). Regular monogyny is a switchof monogynous colonies to polygyny by fusing colonies (usually observed in polygynous species,cf. Hölldobler & Wilson 1990) when suitable nest sites are scarce; this phenomenon wasobserved in the usually monogynous species Leptothorax (Myrafant) nylanderi (Förster, 1850)(cf. Buschinger 1967, 1968, 1974). Fusion of colonies in this species was observed also by Foitzik& Heinze (1998, 2000); however, polygyny is only temporary and results in intraspecific slavery(Foitzik & Heinze 1998, 2000) – one of the queens is subsequently eliminated, and workers of hercolony serve in the alien colony.151

Study of the above phenomene is important for understanding the role of kin-selection andother factors shaping the evolution and maintenance of eusociality.The existence of monogynous/polygynous strategies in Leptothorax (Myrafant) slavonicus, aspecies only recently (Seifert 1995) separated from L.(M.) nylanderi, has not been studied. Ourinvestigations began with a study of populations of “L. (M.) nylanderi” from Bohemia and Slovakia.As preliminary observations (Kašpárek unpubl.) suggested, and subsequent field and laboratorystudies confirmed (Tichá 1992) these populations are less tolerant of several queens in acolony than West-European populations (Donisthorpe 1927, Chauvin 1947, Buschinger 1967,1968). Seifert (1995, 1996) studied the morphology of East-European populations of “L. nylanderi”,and concluded (supported by genetic research carried out at the University of Lund-Douwespers. comm. in Seifert 1995) that the populations on the eastern side of the line Schwerin–Magdeburg–Leipzig–Döbeln (Germany) belong to an unrecognised parapatric species, Leptothoraxslavonicus. It was first assigned subspecific rank, Leptothorax nylanderi slavonicus in 1995,and that of a distinct species in 1996. The populations we studied belong to this species.This discovery may account for the differences in behaviour of Czech, Moravian and Slovakpopulations of the above species, and have stimulated other ethological and sociobiologicalstudies of L. slavonicus, a species not previously studied in this way (except the seminar study ofTichá 1992).MATERIAL AND METHODS1. Terminology (see the Introduction as well)Queen – dealate, fully fertile, egg-laying female, parent of next generations of females, strongly attractive forher nestmates, dominant in the colony (in Leptothorax slavonicus the queen is located in a centre of the colonyon a heap of brood, and does not take part in the labour);worker (in L. slavonicus) – wingless female; if egg-laying, then producing non-fertilised eggs only; co-operatingwith nestmates and performing basic tasks necessary for the well-being of the colony and maintenance of thenest;brood – juvenile stages;orphaned colony – colony lacking a queen;regulation towards monogyny – reduction in the number of queens in a colony resulting in monogyny;recruitment – increased motor activity of some individuals, which touch the other ants with their antennae andlegs, leading to their mobilisation;tugging – mutual pulling in opposite directions by two (or more) workers, or pulling the queen by a group(groups) of workers;accepted queen – the female chosen as a queen after fusion of colonies and fully satisfying the above criteria ofa queen;neutral formicarium – clean formicarium uninhabited before the experiment.2. Origin of the colonies used in this studyLeptothorax slavonicus – 238 colonies were collected in June to September over the period 1989–2000.Czech Republic – 176 colonies: Mlník env. (14° 32’ E, 50° 28’ N, Nat. grid No 5553 [after Pruner & Míka1996]); Praha (14° 22’ E, 50° 03’ N, No 5952); Mohelno env. (16° 11’ E, 49° 07’ N, No. 6863), Znojmo env.(15° 54’ E, 48° 59’ N, No. 7061). Slovakia – 62 colonies: Piešany env. (18° 53’E, 48° 18’ N, No. 7769) (Fig. 1).Colonies of this small ant usually contain only several dozen individuals, and inhabit small cavities, mainlyhollow, dead tree twigs and acorns. They were collected mainly in deciduous (Quercus, Carpinus), mixed and pinewoods. Complete colonies, mostly still in their nest-sites, were placed into plastic bags (when this was notpossible, the ants were collected in vials by using an aspirator), and then transferred to the laboratory.3. CulturesNumber of individuals (queens, alate females, workers, males, brood) in each colony was counted, and the antswere then transferred into tripartite plastic formicaria. Here they were bred (according to Buschinger 1967) andmanipulated. Dimensions of the formicaria: cylindrical nest-site, r – 75 mm, l – 135 mm; three types of arenas– 120×80×30 mm, 170×90×70 mm, 270×90×70 mm – each type was used for ten experiments of Series 1 (see152

elow) and a 170×90×70 type was used in the Series 3 experiments. For the Series 2 experiments an arena sized120×80×30 mm was used; the flat glass chambers (dimensions 20×20×3 mm) with a millimetre grid were used asthe nest-site – this arrangement facilitated observation of ant movements.4. Marking antsAll the queens in Series 1 were marked by amputation of the last tarsal segment of some of the legs, the queens andthe workers used in the Series 2 experiments were marked with acetone nail varnish applied to the thorax orabdomen (manipulation during marking ants as described by Heinze 1993, 1996).5. ExperimentsSERIES 1. Two monogynous colonies (usually of different size) were placed in the arena of a neutral formicarium.Behaviour of the ants was monitored visually, continuously for the first 4 hours, over the next 10 hours at 15minutes intervals, and than at intervals of 6 hours up to the termination of the experiment. This experiment wasreplicated 30 times and each lasted 6 weeks. Questions addressed: When nest-sites are scarce do colonies ofLeptothorax slavonicus fuse (as described for L. nylanderi by Buschinger 1967, 1968 and Foitzik & Heinze 1998,2000)? If they do, does queen regulation occur?SERIES 2. Ten workers and their queens were marked and used instead of the colonies in experiment identical withSeries 1 experiments. There were 5 replicates of this experiments. Timing of the observations was as in Series 1.Duration of each experiment was 6 weeks. Questions addressed: Does the original nestmates continue torecognise one another, and does any hierarchy develop in the mixture?SERIES 3. Two foreign queens were added to an orphaned colony. This was repeated ten times. Observations wereas in Series 1 and 2. Duration of each experiment was six weeks. Questions addressed: What is the behaviouralresponse of orphaned colonies to foreign queens and is there any regulation towards monogyny when foreignqueens were adopted?6. Statistical assessment of resultsFig. 1. Localities in the Czech Republic (CZ) and Slovakia (SK) where the colonies of Leptothorax slavonicusSeifert used in the experiments were collected.153

Tab. 1. Summary of experimental results. Explanations: N – number of experiments, NRM – number of regulationsto monogyny (= elimination of supernumerary queens), NF – number of fusions of two monogyous colonies,N2M – number of originations of two monogynous colonies, N0 – number of experiments resulting in orphancyof the colony, NAQ – number of adoptions of an alien queen; * – adoption before orphancy, ** – adoption of awinning by workers from the colony of the eliminated queen after the fusiondesign N = 100% NRM NF N2M NO NAQseries 1 30 24 24 6 1* 24**series 2 5 0 0 5 0 0series 3 10 6 0 0 4 6Fisher exact test from Statistica 93 was used in the statistical evaluation of the choice of a queen.RESULTS1. Monogyny of Leptothorax slavonicus in the fieldAll of the 238 colonies examined (100%) had only one queen.2. Experiments (Tab. 1, Fig. 2)SERIES 1. The mixing of two monogynous colonies (30 experiments) resulted in the colonies fusingto form one colony, and the workers reducing the number of queens to one in 24 cases (monogynouscolonies formed in all cases except one, in which the accepted queen died before the terminationof the experiment, and an orphaned colony resulted). The colonies separated and formed twomonogynous colonies, each in a different part of the formicarium in 6 cases. A digynous colonywas never formed.SERIES 2. The mixing of two marked groups of ants (5 experiments) resulted in the ants of thedifferent colonies living in one formicarium, but as two autonomous and distinct fractions. No caseof fusion was observed.SERIES 3. When two foreign queens were offered to an orphaned colony (10 experiments) onequeen was adopted, the other eliminated in 6 cases, and both queens eliminated, i. e. the coloniesremained without queen, in 4 cases.3. Behaviour of ants during experiments3. 1. Experimental Series 1The ants first explored the arena and nest-site, which was accompanied by aggressive interactionsbetween the workers and attacks on both queens, then formed a cluster around each queen, andeither retained this separation up to the termination of the experiment, or the clusters inhabited thenest-site, formed a fused colony, and regulated the number of queens to one. Six successivebehavioural phases (unless stated otherwise) were identified in nearly all the experiments : a-b-cd-e-f,or a-b-c-d-g-h.(a) Exploration. During this phase the ants became acquainted with the new situation, arena andstrangers. This was associated with increased mobility with exploratory behaviour prevailing overother activities. Ritualised attacks in which the mandibles were opened wide against strangers,but there was no body contact, were common. Grooming of the queens by workers lasted until oneof the queens was eliminated. Duration of this phase was about 15 minutes.(b) Aggregation of workers around the alien queen during which the workers seized the legs,antennae, head of the queen and dragged it across the arena or transported it by holding it by itspetiole and attacked it with their mandibles. The queens characteristically remained passive andrigid during transportation. At the same time the workers explored the nest-site and transported154

the brood, first into heaps near the shaded part of the arena and later into the nest-site, andtransported their queens to the heaps of brood. Trophallaxis among workers was observed untilthe experiment was terminated. Both queens were fed prior to regulation, when only the acceptedone was fed. In cases when the colonies separated, feeding of both queens continued. Recruitmentoccurred in 18 experiments. Individual conflicts took the form of tugging (27 experiments)and fighting (24 experiments); group conflicts also involved tugging (15 experiments) and fighting(7 experiments), but rarely stinging (3 experiments). In two cases both the queens used theirsting in mutual fights that lasted 20 or 40 seconds (no evident relation with any other factor).Duration of this phases was 10 minutes to 2.5 hours.(c) Formation of two monogynous colonies occurred when the brood was divided into two heaps,each of which was occupied by one of the two queens accompanied by a group of workers. Thiswas followed by the subdivision of the remaining workers in two groups. Duration of this phasewas 10 minutes to 2.5 hours.(d) Decreased activity occurred when the ants in both groups remained immobile for 25 minutes to24 hours.The next two successive phases occurred in one of the two variants.VARIANT 1 (e, f)(e) Occupation of the nest-site by both the colonies. This was accomplished by the movement ofindividual ants, or their passive transport in the supine position (head or the petiole held by otherants) along with the transfer of both the queens. The resulting colony was seemingly digynous,however egg-laying by the queens was never observed. This took 23 hours to 22 days to complete.(f) Return to monogyny by the elimination of one of the queens at the periphery of the arenaoccurred in 24 of 30 experiments, usually within 23 days of the start of the experiment. This wasFig. 2. Results of experiments series 1–3. Explanations: N – number of experiments, NRM – number of regulationsto monogyny (= eleimination of supernumerary queens), NF – numbers of fusions of two monogynouscolonies, N2M – number of originations of two monogynous colonies, N0 – number of experiments resulting inorphancy of the colony, NAQ – number of adoptions of an alien queen.155

achieved by workers attacking one of the queens with their mandibles, never with their stings.The eliminated “queen” died while being transported (3 out of 24) or immediately after (13 out of24), or was kept in isolation at the periphery of the arena, without grooming and trophallaxis (8 outof 24). The workers action was co-ordinated, with no mutual conflicts, and the rejected “queen”remained passive. The accepted queen was adopted by the workers and monogyny persisted untilthe termination of the experiment. This phase took between 10 minutes to 8 hours to complete.VARIANT 2 (g, h)(g) Occupation of the nest-site by one colony while the other remained in the arena, which wasaccepted as a “nest-site” on 6 occasions. This phase lasted for between 10 minutes and 4 hours.(h) Return to monogyny. The two colonies functioned independently and behaved as two distinctmonogynous colonies until the termination of the experiment.Choice of queenThere was a relation between the actual sizes of the experimental colonies and choice of acceptedqueen (Fig. 3). After fusion of the colonies in the experiments of Series 1 (24 occasions) the queenof the larger colony was chosen in 18 cases, and that of the smaller colony in 6 cases (c 2 = 6, p =0.0143, df = 1).3. 2. Experimental Series 2The behaviour of ants was very similar to that in Series 1, but both the colonies retained theirautonomy and never fused. Conflicts only occurred between members of different colonies, andthe ants were always able to recognise their nestmates (interactions: allogrooming, trophallaxis,etc.) from non-nestmates (interactions: both ritualised and true attacks). The recognition was notimpaired even immediately after marking the ants with nail varnish, the odour of which probablyinterfered with nest scents. The resulting two monogynous colonies were formed exclusively fromthe original nestmates in all five experiments.3. 3. Experimental Series 3These experiments resulted either in adoption of one of the queens or elimination of both, never indigyny. Even in these experiments 5 successive behavioural phases were regularly observedeither in sequence a-b-c-d-e, or a-b-c-d-f.(a) Identification of the queen. Workers approached a queen, touched it with their antennae,departed, etc. This phase lasted about 30 minutes.(b) Aggregation of workers around queens. Workers formed clusters around quite passive queens,seized them, transported and attacked them with their mandibles etc., as in the previous experiments,including the same kind of conflicts between the workers (sting never used), i.e. ritualisedattacks (6 out of 10 experiments), tugging (4 out of 10) and fighting (3 out of 10 experiments), andgroup tugging (1 out of 10) involving maximally 4 individuals. Duration of this phase was 30minutes to 3 hours.(c) Decreased activity. The majority of the workers joined one of the groups around the queens,and remained stationary. This phase lasted far from 55 minutes to 8 hours.(d) Intermediate phase with two “queens”. Formation of a seemingly digynous colony in the nestsite,but the queens did not lay eggs. Duration of this phase was from 5 hours to 6 days.VARIANT 1(e) The establishment (6 experiments out of 10) of monogyny. One of the two “queens” in thecolony was attacked in the nest-site and transferred to the periphery and eliminated. This queendied in 4 cases (out of 6) or was kept in isolation at the periphery of arena without grooming and156

trophallaxis until the termination of the experiment in 2 cases (out of 6 cases) . This occurred notlater than 7 days after the start of the experiment. Duration of this phase was from 30 minutes to 6hours. The remaining queen was adopted and the colony functioned as a monogynous colonyuntil the end of the experiment.VARIANT 2(f) Elimination of both “queens” and return to orphancy (4 experiments out of 10). Within 6 daysboth “queens” were attacked, eliminated and died. This phase lasted far from 3 hours to 2 days.The orphaned colony remained in this state up to the end of the experiment.DISCUSSIONBoth field observations and laboratory experiments have shown that Leptothorax slavonicus isan obligatory monogynous species, as are many other species of the subgenus Myrafant Smith,1950. The regulatory mechanisms restor monogyny reliably in cases when in a colony occur twoqueens. Monogynous colonies may fuse, regulate number of queens and form genetically heterogeneouscolonies. This occurs in Leptothorax nylanderi (cf. Foitzik & Heinze 1998, 2000) whereit is regarded as intraspecific slavery.Frequent finds of wood cavities resembling empty nest-sites of L. slavonicus in winter (Kašpárek& Tichá unpubl.) may suggest colony fusion occurs in this species under natural conditions.Hence we cannot exclude existence of intraspecific slavery in this species. Moreover, our experimentsindicate that nest-site limitation plays a role in the fusion of colonies of L. slavonicus (as inL. nylanderi) – first the two colonies attempt to live independently, but after some time elapsesthey occasionally fuse.However, L. slavonicus seems less tolerant than L. nylanderi of even the temporary presence ofseveral queens in a colony. This is supported by a faster reduction in the number of queens thanin the latter species. Foitzik & Heinze (1998) recorded the survival of several “queens” in onecolony of L. nylanderi for more than 64 days, while in our experiments on L. slavonicus theregulation towards monogyny took place within 23 days.Fig. 3. Choice of the queen. Explanations: NE – number of experimental colonies, NMC – number of members ofthe colony; negative values = the winning queen from smaller colony.157

This difference was observed by Kašpárek (pers. comm.) and Tichá (1992) prior to the taxonomicseparation of the two species, and we then hypothesised that “L. nylanderi” consists of two ormore genetically and ethologically distinct populations. Independently of this hypothesis, thiswas proved by morphological and genetic studies, which revealed it consisted of two parapatricspecies. The West European L. nylanderi (type locality Aachen, Germany fide Radchenko 2000)and the Central and East European L. slavonicus (type locality Kr. Görlitz, Schönau-Berzdorf,Hutberg; cf. Seifert 1995, Radchenko 2000). The uncertainty of several authors (Donisthorpe 1927,Chauvin 1947, Buschinger 1967, 1968, Foitzik & Heinze 1997, 1998, 2000, Plateaux, pers. comm.)concerning monogyny/polygyny and monoandry/polyandry of colonies of West European populationsof L. nylanderi (Chauvin 1947, Plateaux 1970, 1978, 1981, Foitzik et al. 1997) indicate theneed for morphological and genetic studies of metapopulations of this species in regions not yetinvestigated and to re-investigate its ethology.The second series of experiments showed that the workers of L. slavonicus consistently recognisenestmates from non-nestmates, even under conditions that probably impair recognition ofnest scent, in this case the presence of acetone vapours. Nestmate recognition in ants is based onpresence of nest-specific odour which may be individually borne or collectively shared and transferred(Gestalt) (Crozier & Dix 1979). This odour involves discriminators recognisable by all themembers of a colony, e. g. cuticular hydrocarbons (e. g. Bonavita et al. 1996, 1997, Meskali et al.1995a, b), polar cuticular lipids (Franks et al. 1990), food quality (Jutsum et al. 1979), a nest material(Breed et al. 1995, Heinze et al. 1996) etc. Leptothorax (Myrafant) lichtensteini Bondroit, 1918, aspecies closely related to L. slavonicus, has nestmate recognition probably based on a Gestalttype model (Provost 1985, 1989, 1990, Provost et al. 1993). Strong influence has a nest material oncolony odour in another related species, Leptothorax nylanderi (Heinze et al. 1996). This factormay explain the ease with which colonies of this species fuse (Foitzik & Heinze 1998). The natureof the recognition in L. slavonicus is still unknown. However, its ability to recognise nestmatesconfirms the general opinion (see, e. g., Holldöbler & Wilson 1977) that monogynous speciesshow a higher degree of closeness than polygynous ones.The marked individuals from different colonies did not mix, probably because of the smallnumber of individuals involved, and hence the large amount of normal space available. Consequently,we could not determine whether nestmate recognition persists for a long period aftercolony fusion or whether a hierarchy associated with colony origin exists in mixed colonies.The series of third experiments showed that orphaned workers of L. slavonicus readily adopt analien queen. Although this has previously been recorded by Foitzik & Heinze (1998) in L. nylanderi,it is strange, because the presence of a queen in leptothoracine ants usually inhibits thelaying of unfertilised eggs by workers, which is rather common in orphaned colonies (Heinze1997). The situation seems to be in conflict with the basic tenets of kin selection: there should exista conflict of interests between the workers and the queen concerning maternal origin of males(Hamilton 1964, Trivers & Hare 1976). An important task for future research is to determine whetherthe inhibition of egg-laying by workers occurs in the presence of an adopted alien queen.CONCLUSIONS1. The species Leptothorax slavonicus is monogynous; no case of polygyny was found in naturalor experimental conditions.2. The maintenance of monogyny was accomplished by behavioural regulation which resulted inthe elimination of supernumerary queens by workers.3. Recognition of nestmates occurred even under conditions of impaired scent communication.158

4. The queen that was accepted was usually that of the larger of the two colonies used in eachexperiment.5. Mixing two monogynous colonies usually resulted in their fusion, formation of a mixed colonyand reduction in the number of queens (monogyny).6. Genetically heterogeneous colonies may persist under experimental conditions.7. In the laboratory orphaned colonies will adopt an alien queen.ADDENDUMLeptothorax (Myrafant) slavonicus is probably a junior subjective synonym of Leptothorax (Myrafant)crassispinus Karawajev, 1926, a species described originally from Russia (Radchenko 2000,Seifert pers. comm., October 2001).A c k n o w l e d g e m e n t sThe initial and final phases of this study were carried out in the Department of Zoology, Charles University,Prague; we gratefully acknowledge the financial support of this Research project by the Ministry of Schools andEducation of Czech Republic Grant No. J 13/98113100004. We are obliged to R. Kašpárek (Praha) for continuousadvice, assistance in maintenance of cultures and carrying out the experiments and to A. Buschinger (Darmstadt),D. Frynta (Praha), L. Plateaux (Nancy), E. Provost (Marseilles), B. Seifert (Görlitz), P. Werner (Praha), J.Žárek (Praha) for valuable information.REFERENCESBONAVITA-COUGOURDAN A., RIVIERE G., PROVOST E., BAGNERES A.-G., ROUX M., DUSTICIER G. & CLEMENT J.-L.1996: Selective adaptation of the cuticular hydrocarbon profiles of the slave-making ants Polyergus rufescensLatr. and their Formica rufibarbis Fab. and F. cunicularia Latr. slaves. Comp. Biochem. Physiol. 113B: 313–329.BONAVITA-COUGOURDAN A., BAGNERES A.-G., PROVOST E., DUSTICIER G., & CLEMENT J.-L. 1997: Plasticity of thecuticular hydrocarbon profile of the slave-making ants Polyergus rufescens depending on the socialenvironment. Comp. Biochem. Physiol. 116B: 287–302.BREED M. D., GARRY M. F., PEARCE A. N., HIBBARD B. E. & RAGE R. E. J. 1995: The role of wax comb in honey beenestmate recognition. Anim. Behav. 50: 489–496.BUSCHINGER A. 1967: Verbreitung und Auswirkungen von Mono- und Polygynie bei Arten der Gattung LeptothoraxMayr (Hymenoptera, Formicidae). Unpubl. Inaugural-Dissertation, Würzburg, 115 pp.BUSCHINGER A. 1968: Mono- und Polygynie bei Arten der Gattung Leptothorax Mayr (Hymenoptera, Formicidae).Insectes Soc. 15: 217–226.BUSCHINGER A. 1974: Monogynie und Polygynie in Insektensozietäten. Pp.: 862–896. In: SCHMIDT G. H. (ed.):Sozialpolamorphismus bei Insekten. Stuttgart: Wiss. Verlagsges MBH, xxiv+974 pp.CHAUVIN R. 1947: Sur l’elevage du Leptothorax nylanderi (Hymenoptere Formicide) et sur l’essaimage in vitro.Bull. Soc. Zool. Fr. 81: 151–165.CROZIER R. H. & DIX M. W. 1979: Analysis of two genetic models for the innate components of colony odor insocial Hymenoptera. Behav. Ecol. Sociobiol. 4: 217–224.DONISTHORPE H. 1927: British Ants. Their Life-history and Classification. London: G. Routledge & Sons, 436 pp.FOITZIK S., HABERL M., GADAU J. & HEINZE J. 1997: Mating frequency of Leptothorax nylanderi ant queensdetermined by mikrosatelite analysis. Insectes Soc. 44: 219–227.FOITZIK S. & HEINZE J. 1998: Nest site limitation and colony takeover in ant Leptothorax nylanderi. Behav. Ecol.4: 367–375.FOITZIK S. & HEINZE J. 2000: Intraspecific parasitism and split sex ratios in a monogynous and monandrous ant(Leptothorax nylanderi). Behav. Ecol. Sociobiol. 47: 424–431.FRANKS N. R., BLUM M.S., SMITH R. & ALLIES A.B. 1990: Behaviour and chemical disguise of cockoo ant Leptothoraxkutteri in relation to its host Leptothorax acervorum. J. Chem. Ecol. 16: 1431–1444.HAMILTON W. D. 1964: The genetical evolution of social behaviour. J. Theor. Biol. 7: 1–52.HEINZE J. 1993: How to immobilize an ant? Insectes Soc. 40: 231–232.HEINZE J. & BUSCHINGER A. 1986: Queen polymorphism in a non-parasitic Leptothorax species (Hymenoptera,Formicidae). Insectes Soc. 34: 1, 28–43.159

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