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HNAN Abundance (s 105 L- 1) 0 0 50 100 150 200 I 2 3 4 Depth (m) 5 6 7 8 9 10 Figure 3.12 - HNAN abundance (x 105 L-) for three detailed study lakes. Ace Lake (a), Pendant Lake (b) and Triple Lake (c) were all depth sampled over several sampling dates. Deep Lake and Club Lake are not included because virtually no flagellates were observed in the samples. Ace Lake sampling date key: 14/01/00 (-"-), 04/03/00 15/07/00 ( ), 07/09/00 () and 15/11/00 ( ). Pendant Lake sampling date key: 23/02/00 (-"-), 17/07/00 (-s ), 07/09/00 () and 13/11/00 ( ). Triple Lake sampling date key: 23/01/00 (-4-), 17/07/00 (-s-), 07/09/00 () and 10/11/00 ( ). 0 PNAN Abundance (s 105 L- 1) 0 500 1000 1500 2 3 4 Dept15 (m) 6 7 ýJ- 9 10 Is Figure 3.13 PNAN - abundance (x 105 L-') for three detailed study lakes. Ace Lake (a), Pendant Lake (b) and Triple Lake (c) were all depth sampled over several sampling dates. Deep Lake and Club Lake are not included because no flagellates were observed in the samples. Ace Lake sampling date key: 14/01/00 (-"-), 04/03/00 (- -), 15/07/00 ( ), 07/09/00 () and 15/11/00 ( ). Pendant Lake sampling date key: 23/02/00 (-"-), 17/07/00 (-"-), 07/09/00 () and 13/11/00 ( ). Triple Lake sampling date key: 23/01/00 (-"-), 17/07/00 (- -), 07/09/00 () and 10/11/00 ( ). 80
3.3 - Discussion 3.3.1 - Summer sampled lakes The forty one lakes investigated displayed a wide range of characteristics. including extremely variable salinities, temperatures and inorganic nutrient conditions. The majority of these lakes have been sampled to some extent in the past as shown in the literature (Table 3.2). Seven of the sampled lakes are not mentioned in any available literature, these are Trident, Pointed, Calendar, Hand, Nicholson, Medusa and Cemetery. Ice cover was assessed in all the lakes. During the summer (1999/2000) ambient temperatures ranged from -6.2°C to 3.3°C (Tables 3.1 a, b and c). Sustained periods of ambient temperature above zero combined with high incident solar radiation caused ice melt leading to partial or complete loss of ice cover. Saline and hypersaline lakes (Tables 3.1 a and b) showed a large reduction in the extent of ice cover or complete loss of cover (e. g. Oval, Lebed, and Cemetery, see table 1 a). Salinity alters the freezing point of water (Williams and Sherwood, 1994), for example sea water has a salinity of 3.8% or 38ppt and does not freeze until the temperature falls to -1.9°C. This is because salinity increases the solute potential and allows better thermal conductivity within water (Williams and Sherwood, 1994), therefore extremely saline water will heat up or cool down much faster than freshwater (Ferris & Burton, 1988). Hence, the saline (4-35ppt) and hypersaline (>35ppt) lakes (Tables 3.1a and b) are more prone to melt out during `warm' spells in the Antarctic summer (Franzmann, 1991; Gore, 1996a). Oval Lake at the time of sampling had -85% ice cover and a water temperature of -1.9°C: this temperature was probably an isolated event due to a combination of high thermal conductivity (salinity) and loss of thermal insulation (ice cover). Ice cover has an extensive impact on the water chemistry of a lake. Maintenance of ice cover throughout most of the year as seen in many of the freshwater lakes (Laybourn-Parry et al., 1992; Laybourn-Parry & Marchant, 1992b; Bayliss & Laybourn-Parry, 1995) isolates the water body from most external influences, such as wind induced mixing (Laybourn-Parry et al., 2001 a) and allochthonous input of, for example animal faeces and melt water (Bell & Laybourn-Parry, 1999a). Surface water temperatures (Table 3.1 a, b& c) are directly dependant on air temperature where ice cover is lacking. Periods of maintained high air temperature lead 81
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COLD ADAPTATION STRATEGIES AND DIVE
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, 71 "If you march your Winter Jour
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1.6.3. Molecular typing techniques
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3.2.1.2.3. DOC and chlorophyll a ..
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5.3.2.5. The Sphingomonas group, is
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LIST OF FIGURES Figure I. I. Diagra
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Figure 3.17a Mean bacterial abundan
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LIST OF TABLES Table 1.1. Types of
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List of Abbreviations AFP AFGP AFLP
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ACKNOWLEDGEMENTS This study was fun
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Chapter 1: INTRODUCTION 1.1 - Tempe
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1999 ; Hand & Burton, 1981) or iden
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Cold adaptation is dependant on the
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not at 25°C is a function of the a
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The most prolific literature on col
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process known as thermal hysteresis
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dominant, role in the ice-growth in
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explanation for this diversity is t
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1.5.3.5 - Membrane bound solute tra
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een well established (Russell & Nic
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& Bowman, 2001; Labrenz & Hirsch, 2
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The reaction mix contains both deox
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gene mixtures and hence can be used
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displaying a high number of common
Page 49 and 50: AACGTCGAAA AACCTCGAAA (Organism A)
Page 51 and 52: particular set of environmental par
Page 53 and 54: Lake Maximum Depth (m) Approximate
Page 55 and 56: Larsemann Hills __ rp3rtrnent of th
Page 57 and 58: Sampling trips took two days during
Page 59 and 60: Sigma, Sigma-Aldrich Company Ltd.,
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Page 65 and 66: min to ensure complete extension of
Page 67 and 68: Figure 2.3 - Lane delineation image
Page 69 and 70: Figure 2.6b - Molecular weights are
Page 71 and 72: RNA 41F (5'-GCT CAG ATT GAA CGC TGG
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Page 79 and 80: information regarding the cold adap
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Page 83 and 84: M N N 't M M \p \p 00 kf) M kf) bA
Page 85 and 86: as would normally be expected, as b
Page 87 and 88: 1444 6ý L CC vý O Q O O NO u 'c z
Page 89 and 90: 5 - 250 3 245 240 G (, -3 235 a 0-
Page 91 and 92: 3.2.2.2.5 - Dissolved organic carbo
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Page 95 and 96: SRP ('g L- 05 10 15 20 2 3 Depth4 5
Page 97 and 98: 3.2.2.3 - Biological characteristic
Page 99: a 0 Bacterial Abundance (x 106 ml-1
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Page 105 and 106: chemocline (Rankin et al., 1999). M
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Page 109 and 110: The temperature (Fig 3.2a) through
Page 111 and 112: et al., 1999). No melt out was obse
Page 113 and 114: increase in ammonium is probably du
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Page 117 and 118: main pigments were chlorophylls a a
Page 119 and 120: Inorganic nitrogen (NO2, NO3 and NH
Page 121 and 122: 744 -- 7 644 6 19 ü SE 400-- 4 2s
Page 123 and 124: salinity decreased from July to Sep
Page 125 and 126: a Bacterial abundance (x 106 1.1) 0
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Page 129 and 130: undoubtedly true of Club Lake. The
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Page 133 and 134: Pendant Lake is an unstratified, sa
Page 135 and 136: non-AFP active peptides from inhibi
Page 137 and 138: . ter rý :. 'ý Figure 4.2 -Fryka
Page 139 and 140: -ý- ---- ,. r. __-_-. ___. r. ýý
Page 141 and 142: 4.2 - Results 4.2.1 - Sampling and
Page 143 and 144: SPLAT score. It has been shown that
Page 145 and 146: 3'E t ý _d M! .PJ r rn 15 3.5 4 3
Page 147 and 148: 4.3 - Discussion Of the 866 bacteri
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Chapter 5: BACTERIAL MOLECULAR TYPI
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Fig 5.1 - Photographs of ARDRA gel
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Coefficient: Dice Algorithm: lPGMAA
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The closest published relative for
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identification was made based on 15
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Isolate number Closest phylogenetic
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e included due to its small sequenc
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5.3.2 - Identification of the AFP a
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explains the fluctuation in related
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contamination was lower as the PCR
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seen to a certain extent in the ARD
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American Type Culture Collection (A
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Phylogenetic cluster alignment of t
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et al., 1994) and the Antarctic SIM
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Chapter 6: BACTERIAL COMMUNITY ANAL
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6.2.1 - Detection of AFP active iso
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using the same PCR protocol as orig
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Figure 6.1 - Denaturing gradient ge
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6.2.2 - Temporal and spatial variat
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Figure 6.4c - Gel I image showing b
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Coefficient: Dice Algorithm: UPGMA
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etween Om and 2m was obviously the
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All PCR-based rRNA molecular techni
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6.3.2.3 - Cellular lysis and DNA pu
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hybrids should be equal. However, S
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contain contaminants. If the profil
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(Section 6.3.2.4.3 ) could not be r
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most dominant species in the commun
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study was characterised as M protea
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11 species) showed AFP activity. Th
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that AFP activity could have evolve
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7.2 - Future Work The 19 AFP active
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REFERENCES ABYZOV, S. S. (1993). Mi
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BELL, E. M. and LAYBOURN-PARRY, J.
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Symposium on Environmental Biogeoch
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Research. 5,477-493. CLESCERI, L. S
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DRICKAMER, K. (1999). C-type lectin
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antifreeze proteins from Atlantic s
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FRANZMANN, P. D. and DOBSON, S. J.
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GRIFFITH, M., ALA, P., YANG. D. S.
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IVANOVA, E. P., KIPRIANOVA, E. A.,
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Antarctic Ecosystems. G. A. Llano (
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Howard-Williams and I. Hawes). Balk
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W. (1998). Rep-PCR-mediated genomic
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putida strains from Antarctica. Mic
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NICHOLS, D., BOWMAN, J., SANDERSON,
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PEARCE, D. A. and BUTLER, H. G. (20
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(1999). Palaeohydrological modellin
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tracers of biogeochemical processes
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R. G. Landes Company, Austin, Texas
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partial 16S rDNA sequencing. Applie
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YABUUCHI, E., WANG, L., ARAKAWA, M.
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APPENDICES Al. Media Tryptic Soya A
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"D C 'O I. .. r .. r 6 rr E , 4mo .
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. + + + I I I I I I + I + + I I I I
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Ö ON N M 1 O - N 'zt "o l'" 00 C N
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1 T 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
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1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
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1 1 1 1 I 1 + I I 1 I 1 1 I 1 1 1 I
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1 1 1 1 1 1 1 I I 1 I 1 I 1 1 I T I
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Cl M 'tt tn '. p t- 00 O\ N M O - (
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d' O - (N M 'nt V) \O N. 00 O> N M
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1 1 T 1 1 1 T 1 1 1 1 1 1 1 1 1 1 1
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+ + i i i i i i " i " . + + + + " +
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e O --ý N M t Vn 1,0 [-- 00 O> N M
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FT ce + I I r. to \, O t- 00 O\ N M
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kn 110 r- 00 Oý N M O - N M Itt vn
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, It N M "t N M Iýt N M M M M Oý
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gj a - - - - - - - - - - - - - - -
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