Lactate dehydrogenase from the Antarctic eelpout, Lycodichthys dearborni

As part of our studies to examine the molecular basis of cold-adaptation, we have determined the kinetic properties, thermal stability and deduced amino acid sequence of the enzyme lactate dehydrogenase (LDH) from an Antarctic zoarcid fish, Lycodichthys dearborni. Unlike Antarctic notothenioid fish which are endemic to the Southern Ocean, zoarcid fish are cosmopolitan and have a substantially longer evolutionary history as a sub-order. The A₄-LDH isoform was isolated and purified from the white muscle of L. dearborni. The kinetic parameters K_m^PYR and k_cat were determined at temperatures from 0 to 25°C. K_m^PYR was substantially higher at low temperatures than those from Antarctic and temperate notothenioid fish, whereas k_cat at these temperatures was essentially the same as those of the other fish LDH in this study. The sequence of L. dearborni A₄-LDH was determined from cDNA derived from white muscle RNA and found to be similar to, but distinct from, the A₄-LDH sequences of Antarctic notothenioid fish. Molecular modelling based on the structure of the A₄-LDH from Pagothenia borchgrevinki suggested that three conservative amino acid changes within the core of the protein that are not directly part of the active site but which might nonetheless influence the active site, may be important in cold-adaptation in L. dearborni A₄-LDH, and that several other changes on the surface of the protein might also play a role in cold-adaptation.     

Diet of Antarctic fur seals Arctocephalus gazella at the Danco Coast,Antarctic Peninsula

The diet of non-breeding male Antarctic fur seals, Arctocephalus gazella, was investigated at the Danco Coast, Antarctic Peninsula, by the analysis of 31 and 149 scats collected from January to March 1998 and 2000, respectively. Overall, fish and krill, followed by penguins and squids, were the most frequent prey and constituted the bulk of the diet. The importance of the remaining taxa represented in the samples (octopods, gastropods, bivalves, isopods, polychaetes and poriferans) was negligible. Among fish, channichthyids constituted the bulk of the diet, with Chionodraco rastrospinosus and Chaenodraco wilsoni, followed by the nototheniid, Pleuragramma antarcticum, being the main prey. The myctophid, Electrona antarctica, was the most frequent and numerous fish prey. The results are discussed and compared with those reported for the South Shetland Islands, the closest area for which similar information is available.     

A first attempt at determining larval growth in three Antarctic fish from otoliths and RNA/DNA ratios

Otolith growth and RNA/DNA ratios of larval stages of Notolepis coatsi, Gobionotothen gibberifrons and Trematomus scotti, three Antarctic fish, were studied during early 1998. RNA/DNA ratios were significantly different between the notothenioids and paralepidids (P<0.01), but similar among notothenioids (P>0.05). The ratios were independent of larval total length and water temperature. Recent otolith growth (based on the last five increments) and biochemical indices correlated slightly, but not significantly. N. coatsi inhabited deeper and colder waters of the Weddell Sea and showed less growth than the other species, which are associated with the Antarctic Peninsula. In all three species, some larvae had very good growth rates though most were rather poor. Recent growth indices might allow the detection and back-dating of growth changes in Antarctic larvae, and provide insight into a crucial yet poorly understood life-phase of these fish.     

The role of iceberg scours in niche separation within the Antarctic fish genus Trematomus

Species of the Antarctic fish genus Trematomus occupy different trophic niches. It is not clear, however, whether small-scale variability in benthic community structure affects niche separation. Therefore abundance and biomass of fish were determined and stomach content and food composition were compared in areas affected by iceberg scours and unaffected areas in the Weddell Sea. Trematomus eulepidotus, T. lepidorhinus and T. scotti dominate undisturbed areas, whereas T. nicolai and especially T. pennellii dominate disturbed areas. Total stomach content and number of prey taxa per fish are higher in preferred than in non-preferred areas. These findings indicate that small-scale horizontal patterns caused by iceberg scours play a distinct role in Trematomus niche separation.     

Trace metals in the Antarctic soft-shelled clam Laternula elliptica: implications for metal pollution from Antarctic research stations

The concentrations of Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn were measured in several soft-tissue types of the Antarctic soft-shelled clam, Laternula elliptica, which had been collected from Rothera Research Station, Adelaide Island, Antarctic Peninsula. Concentrations of Mn, Ni, Cr and Pb were significantly higher in the kidney than in any other soft tissue and highest concentrations of Cu were observed in the digestive gland. In general, the total tissue concentrations of heavy metals in L. elliptica were considered to be at baseline levels, except for Cu in organisms near the end of the runway. Copper concentrations were an order of magnitude greater (357 µg/g dry weight) than baseline levels, suggesting anthropogenic contamination or an unidentified natural source. However, there was no indication of anthropogenic metal contamination occurring close to the sewage outfall at Rothera, which is a significant metal source. The results indicate that L. elliptica is a useful long-term biomonitor of heavy metal contamination in Antarctic coastal waters.     

Organization of neuropeptide tyrosine-like immunoreactive system in the brain of the Antarctic fish, Trematomus bernacchii

Antarctic notothenioids have developed unique freezing-resistance adaptations, including brain diversification, to survive in the subzero waters of the Southern Ocean surrounding Antarctica. In this study we have investigated the anatomical distribution of neuropeptide tyrosine (NPY)-like immunoreactive elements in the brain of the Antarctic fish Trematomus bernacchii, by using an antiserum raised against porcine NPY. Perikarya exhibiting NPY-like immunoreactivity were observed in distinct regions of the brain. The most rostral group of immunoreactive perikarya was found in the telencephalon, within the entopeduncular nucleus. In the diencephalon, three groups of NPY-like immunoreactive perikarya were found in the hypothalamus. Two groups of positive cell bodies were found in distinct populations of the preoptic nucleus, whereas the other group was found in the nucleus of the lateral recess. More caudally, NPY immunoreactivity was detected in large neurons located in the subependymal layers of the dorsal tegmentum of the mesencephalon, medially to the torus semicircularis. NPY-like immunoreactive nerve fibres were more widely distributed throughout the telencephalon to the rhombencephalon. High densities of nerve fibres and terminals were observed in several regions of the telencephalon, olfactory bulbs, hypothalamus, tectum of the mesencephalon and in the ventral tegmentum of the rhombencephalon. The distribution of NPY-like immunoreactive structures suggests that, in Trematomus, this peptide may be involved in the control of several brain functions, including olfactory activity, feeding behaviour, and somatosensory and visual information. In comparison with other neuropeptides previously described in the brain of Antarctic fish, NPY is more widely distributed. Our data also indicate the existence of differences in the brain distribution of NPY between Trematomus and other teleosts. In contrast with previous results reported in other fish, Trematomus contains positive fibres in the olfactory bulbs and immunoreactive perikarya in the nucleus of the lateral recess, whereas NPY-immunopositive cell bodies are absent in the thalamus and rhombencephalon, and no NPY immunoreactivity is present in the pituitary. These differences could be related to the Antarctic ecological diversity of notothenioids living at subzero temperatures.     

Comparison of lipid content and fatty acid composition in muscle and liver of two notothenioid fishes from Admiralty Bay (Antarctica): an eco-physiological perspective

During Antarctic summer, total lipids (g/100 g dry matter) in Notothenia coriiceps (n=18) and Lepidonotothen nudifrons (n=10) were low (6.1 and 4.7 in muscle), which is typical of Antarctic benthic species. The liver of female N. coriiceps was heavier and contained more lipids per dry weight than the liver of males. The fatty acid composition of N. coriiceps and L. nudifrons was dominated by polyunsaturated fatty acids (PUFA, respectively, 44 and 49% of total fatty acids in muscle, 31 and 46% in liver), which included primarily C20:5n-3 (18 and 19% in muscle, 13 and 18% in liver) and C22:6n-3 (15 and 19% in muscle, 12 and 20% in liver). In L. nudifrons, the levels of some unsaturated fatty acids increased with age and size. The high percent unsaturation (PUFA+MUFA, 78 and 80% in muscle, 82 and 80% in liver) is a response to low water temperature (-0.4°C). Fish fatty acid profiles reflect fatty acid profiles of the diet (amphipods, macroalgae and fish).     

Triacylglycerol lipase activities in tissues of Antarctic fishes

. The activity of the lipolytic enzyme, triacylglycerol lipase (TAG lipase; E.C. 3.1.1.3) was measured in heart ventricle, liver, oxidative skeletal muscle, and adipose tissue of four species of Antarctic fishes (Chaenocephalus aceratus, Notothenia coriiceps, Trematomus newnesi, Gobionotothen gibberifrons). Estimates of weight-specific activities at 0°C in tissues of the Antarctic fishes were broadly comparable to those reported for temperate-zone fish species assayed at their physiological temperature. Within each species, liver tissue displayed the highest weight-specific activity of TAG lipase in C. aceratus, T. newnesi, and G. gibberifrons, while adipose tissue of N. coriiceps showed the highest activity within that species. Combining these estimates with carefully measured somatic indices of the tissues, permitted assessment of the relative importance of different tissues to overall lipolytic capacity on an organismal level. On this basis, liver tissue displays the greatest capacity of TAG lipase per 100 g body weight in each of the species studied.     

Temperature-dependent pH regulation in stenothermal Antarctic and eurythermal temperate eelpout (Zoarcidae): an in-vivo NMR study

Temperature-dependent adjustments of intracellular pH are thought to play a major role in the maintenance of protein function. Comparative studies were carried out in two species from the same fish family (Zoarcidae), the stenothermal Antarctic eelpout (Pachycara brachycephalum) and the eurythermal eelpout (Zoarces viviparus), to find out whether pH regulation is modified by temperature in the closely related species and to what extent the respective pattern differs between eurytherms and stenotherms. Previous invasive studies had compared individual animals sampled at various temperatures and suggested that a decrease in intracellular pH (pHi) values occurs at rising temperatures, as predicted by the alpha-stat hypothesis of acid-base regulation. The present study used non-invasive in vivo ³¹P-NMR spectroscopy in non-anaesthetized, unrestrained fish for long-term online recordings in individual specimens. Control spectra obtained at T=0°C for P. brachycephalum and at 12°C for Z. viviparus indicated low stress conditions, as well as a high stability of energy and acid-base status over time periods longer than 1 week. Temperature changes had no influence on the concentration of high-energy phosphates like phosphocreatine or ATP. Temperature-induced pH changes were monitored continuously in a range between 0 and 6°C for polar, and 12 and 18°C for temperate eelpout. A pHi change of around -0.015 pH units/°C was observed within both species, in accordance with the alpha-stat hypothesis; however, extrapolation to the same temperature revealed different set points of pH regulation in the two species. These findings confirm that an alpha-stat pattern of pH regulation can be found in stenothermal Antarctic animals, at set points deviating from an alpha-stat pattern, however, in a between-species comparison.     

Effects of UV on the spores of the fungal species Arthrobotrys oligospora and A. ferox

. The effects of UV irradiation (at 5>380 nm and 5>265 nm) on spore suspensions of a European strain of Arthrobotrys oligospora (hyphomycete) have been investigated and compared with those on a strain of the Antarctic species, A. ferox. Emission and excitation spectra of control and irradiated spore suspensions of both strains suggest a higher vulnerability of A. oligospora, which would be consistent with changes in membrane permeability and a lower amount of UV-protecting substances. Germination tests on UV-B irradiated spores confirm the higher resistance of A. ferox, which seems better adapted to the high UV radiation levels characterizing Antarctic environments.     

Parasites of the Antarctic rattail <Emphasis Type="Italic">Macrourus whitsoni</Emphasis> (Regan, 1913) (Macrouridae,Gadiformes)

A total of 386 Macrourus whitsoni from Antarctic waters were examined for ecto- and endoparasites. Sixty-five M. whitsoni collected near Halley Bay (Weddell Sea) and 321 specimens from the continental slope off King George Island (South Shetland Islands) were studied for sphyriid copepods directly after being caught. A subsample of 25 specimens from the Weddell Sea and of 9 specimens from King George Island were studied for the presence of other metazoan parasites. Twenty-two species were found, including one myxozoan, six digeneans, one monogenean, three cestodes, seven nematodes, one acanthocephalan and three crustacean species/taxa. While Auerbachia monstrosa and Capillaria sp. are reported for the first time from around the Antarctic, the other parasites have been recorded earlier in the Southern Ocean. Many parasite species found have a wide zoogeographical range and a low host-specificity. The parasite fauna of M. whitsoni revealed several similarities with its congeners M. carinatus and M. holotrachys from Antarctic and sub-Antarctic waters. This can be explained by a wide host range of many macrourid deep-sea parasites, together with an overlap in distribution patterns of their hosts. Other supporting factors are host migration and a close phylogenetic relationship between the hosts, which enable the parasites to infest all three macrourids. Eight new host and 14 new locality records are established.     

Taxonomic studies of the Antarctic icefish genus Cryodraco Dollo, 1900 (Notothenioidei: Channichthyidae)

Based on a meristic and morphometric study of 101 specimens, we recognise 2 valid species in the Antarctic channichthyid genus Cryodraco: Cryodraco antarcticus Dollo, 1900 and C. atkinsoni Regan, 1914. Although the species overlap in most meristic and morphometric characters, we have distinguished several reliable characters for diagnosis and identification. Multidimensional scaling, a nonparametric multivariate technique, clearly separates the two species on the basis of pelvic fin length, head length, number of second dorsal fin rays and origin of the lower lateral line relative to the anal fin rays. We provide a revised identification key to the species of Cryodraco. From a zoogeographical point of view, C. antarcticus has a circum-Antarctic distribution whereas C. atkinsoni is largely confined to the East Antarctic Zoogeographic Province.     

First record of anomuran and brachyuran larvae (Crustacea: Decapoda) from Antarctic waters

Two decapod crustacean larval morphotypes belonging to the Anomura and Brachyura were found for the first time in Antarctic waters. Nine specimens were obtained from qualitative plankton hauls in Maxwell Bay (Bransfield Strait) (62°14'33S; 58°43'81W) off King George Island, Antarctic Peninsula. The anomuran morphotype belonged to the Hippidae, and apparently to the genus Emerita, whereas the brachyuran representative was assigned to the genus Pinnotheres (Pinnotheridae). At present, species determination is not possible due to lack of knowledge of larval morphology in both families. Adult forms of these reptant decapods are not known from Antarctic waters; the occurrence of the present larval forms is considered as a possible intrusion of Subantarctic water masses into the Antarctic environment. This hypothesis is supported by the additional presence of the copepod genus Acartia in the same sample material, which is exclusively known from Subantarctic waters.     

Feeding strategies and diet composition of four Antarctic cnidarian species

The diet of four species of Antarctic cnidarians, two hydroids and two anthozoans, was investigated. One hydroid, Tubularia ralphii, and one anthozoan, Anthomastus bathyproctus, seem to basically consume zooplankton whereas the other hydroid species, Oswaldella antarctica, has a diet mainly based on the fine fraction of seston. The last cnidarian investigated, the stoloniferan alcyonacean, Clavularia cf. frankliniana, feeds mainly on resuspended material. The wide range of diets of these Antarctic cnidarians indicates opportunistic behaviour by feeding on different sources and taking advantage of the available food sources. Data related to low C/N ratios in the sediment and high NO₂ concentrations in areas with dense communities of benthic suspension feeders, along with data on capture rates, lead us to hypothesize that these organisms play an important role in the recycling processes of organic matter in Antarctic benthic ecosystems.     

<Emphasis Type="Italic">Campylonotus arntzianus</Emphasis>, a new species of the Campylonotidae (Crustacea: Decapoda: Caridea) from the Scotia Sea (Antarctica)

Two specimens of Campylonotus arntzianus sp. nov. were caught in the Antarctic Scotia Sea off Saunders Island (57°40.31'S, 26°27.81'W) using an Agassiz trawl at one station (depth: 475-589 m). The new species described here is the fifth representative of the monogeneric family Campylonotidae, and the first of the family south of the Antarctic Convergence. Campylonotus arntzianus sp. nov. is a shrimp of about 5 cm in total length. Due to similarities in adult morphology, C. arntzianus sp. nov. seems to be closely related to C. capensis, a deep-sea species from the Southern Atlantic Ocean. A simple key for the species identification of the Campylonotidae is provided.     

Microscopic anatomy and ultrastructure of the digestive system of three Antarctic shrimps (Crustacea: Decapoda: Caridea)

Mandibles and stomachs of three Antarctic shrimp species (Nematocarcinus lanceopes, Notocrangon antarcticus, and Chorismus antarcticus) were investigated by means of scanning electron microscopy. Transmission electron microscopy of the midgut glands was applied to find out the nutritional status of the animals, which contained a broad variety of food items in their stomachs. In contrast to the Antarctic krill, the three carideans possess a dual filter system; primary filters in the cardia and secondary filters in the pyloric chamber. Morphologically, the mandibles and stomachs of the three species vary considerably; however, their food items are similar. We conclude that stomach contents do not really reflect the different modes of life or habitat preferences of the investigated species.     

Facing multiple enemies: parasitised hosts respond to predator kairomones

During their lifetime most organisms are exposed to various enemies influencing their victims in multiple direct and indirect ways. Most studies concentrate on the effects of one enemy at a time, thereby not taking into account that in nature organisms are often simultaneously exposed to more than one enemy. We conducted a life-history experiment to investigate the simultaneous effects of predators (fish, Leuciscus idus) and parasites (microparasite, Caullerya mesnili) on their victim (Daphnia galeata). D. galeata were exposed to predator kairomones, parasites or both. D. galeata are able to sense the presence of fish predators via chemical cues (= kairomones). Both fish predator kairomones and microparasite infections influence the life history of Daphnia. Some of the effects of fish predator kairomones are directly opposed to microparasite effects; fecundity, for example, is increased in the presence of fish kairomones and decreased in Daphnia parasitised with C. mesnili. We investigated the influence of both threats on age at maturity, body size at different adult instars, fecundity and survival of one D. galeata clone. In the presence of fish kairomones, all D. galeata matured significantly earlier and increased the number of eggs in the second brood significantly. Parasitised D. galeata matured significantly earlier than non-parasitised ones in the absence and presence of fish kairomones. An infection with the microparasite C. mesnili led to significantly lower clutch sizes at the second adult instar, to significantly smaller body sizes from adult instar three onwards and to significantly reduced survival. No significant interaction effect between the responses to fish presence and to parasite infection was found for any of the investigated life-history traits. The lack of interaction effects between the exposure to predator kairomones and parasite infection was most likely due to the different timing of the effects. Fish kairomones affected D. galeata early in its life history whereas C. mesnili increased in its effects over time. Our results show that parasitised D. galeata are able to exhibit life-history responses to fish predator presence early in their lives. Thus, D. galeata parasitised with C. mesnili have a similar chance as non-parasitised D. galeata to escape from fish predation via life-history changes. Since older parasitised D. galeata are smaller, they may have an even better chance to escape visual predators under actual predation.     

The diet of the South Georgia shag Phalacrocorax georgianus at South Orkney Islands in five consecutive years

The diet of the South Georgia shag Phalacrocorax georgianus at Laurie Island, South Orkney Islands, was studied by the analysis of 420 pellets (regurgitated casts) collected from the 1995 to 1999 breeding seasons. Demersal-benthic fish were by far the main prey, followed by molluscs (mainly octopods and bivalves) and polychaetes. Harpagifer antarcticus (in 1995, 1998, 1999) and Lepidonotothen nudifrons (1996, 1997) were the most frequent prey, followed by Gobionotothen gibberifrons or Trematomus newnesi. Harpagifer antarcticus, followed by L. nudifrons or T. newnesi, was the most numerous prey in all of the seasons and predominated in mass in 1995 and 1996, followed by Notothenia coriiceps and L. nudifrons. In 1997 and 1999, G. gibberifrons and N. coriiceps were the species that most contributed to the diet whereas N. coriiceps and H. antarcticus contributed most in 1998. Our results differ from those reported for the South Orkney Islands in previous studies. These differences could be due to the use of different diet-analysis methods and to shags-related and/or fisheries-related changes in the food availability around the colonies. These alternative explanations are analysed and discussed.     

Chemo-tactile predator avoidance responses of the common Antarctic limpet Nacella concinna

The Antarctic limpet, Nacella concinna, has the ability to escape predation by the predatory sea star, Neosmilaster georgianus. When the sea star comes into contact with the limpet, the limpet undergoes characteristic behaviors that include extension of its pallial tentacles, raising its shell in a mushroom-like fashion, rotation, and flight. These behaviors were not observed until the sea star physically contacted the limpets and were not induced by the omnivorous sea star, Odontaster validus, which occupies the same habitats as Ne. georgianus. Crude hydrophilic extracts from Ne. georgianus, but not from O. validus, induced the escape behaviors, indicating that this response is chemical as well as tactile. These putative escape behaviors displayed by Na. concinna may contribute to avoidance of predation by Ne. georgianus.