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Fourier transform infrared (FTIR) spectroscopy is a biophysical technique used for non-destructive biochemical profiling of biological samples. It can provide comprehensive information about the total cellular biochemical profile of microbial cells. In this study, FTIR spectroscopy was used to perform biochemical characterization of twenty-nine bacterial strains isolated from the Antarctic meltwater ponds. The bacteria were grown on two forms of brain heart infusion (BHI) medium: agar at six different temperatures (4, 10, 18, 25, 30, and 37°C) and on broth at 18°C. Multivariate data analysis approaches such as principal component analysis (PCA) and correlation analysis were used to study the difference in biochemical profiles induced by the cultivation conditions. The observed results indicated a strong correlation between FTIR spectra and the phylogenetic relationships among the studied bacteria. The most accurate taxonomy-aligned clustering was achieved with bacteria cultivated on agar. Cultivation on two forms of BHI medium provided biochemically different bacterial biomass. The impact of temperature on the total cellular biochemical profile of the studied bacteria was species-specific, however, similarly for all bacteria, lipid spectral region was the least affected while polysaccharide region was the most affected by different temperatures. The biggest temperature-triggered changes of the cell chemistry were detected for bacteria with a wide temperature tolerance such Pseudomonas lundensis strains and Acinetobacter lwoffii BIM B-1558.

A novel cold-adapted bacteria Arthrobacter oryzae BIM B-1663 isolated from Antarctic green snow showed keratinase activity and efficient poultry feather degradation. A. oryzae strain degraded more than 80 % of chicken feathers within 7 days of cultivation at 25 °C. The optimal keratinase activity for A. oryzae BIM B-1663 was observed at 50 °C, both for α-keratin (44.86 U/mL) and for β-keratin (94 mU/mL). The obtained results from sulfite and thiol groups tests and Fourier transform infrared spectroscopy (FTIR) showed that A. oryzae strain has a different keratin degradation mechanism than the reference strain Bacillus licheniformis CCM 2145T. FTIR fingerprinting can be used for monitoring of feather hydrolysis as it showed distinct chemical differences in feather meal hydrolysates, retentate and permeate from A. oryzae and B. licheniformis strains.

In this study, for the first time, we report the identification and characterization of culturable fast-growing bacteria isolated from the sea-affected temporary meltwater ponds (MPs) in the East Antarctica area of the Vecherny region (−67.656317, 46.175058) of the Thala Hills Oasis, Enderby Land. Water samples from the studied MPs showed alkaline pH (from 8.0 to 10.1) and highly varied total dissolved solids (86–94,000 mg/L). In total, twenty-nine bacterial isolates were retrieved from the studied MPs. The phylogenetic analysis based on 16S rRNA gene sequence similarities showed that the isolated bacteria belong to the phyla Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes and the twelve genera Pseudomonas, Shewanella, Acinetobacter, Sporosarcina, Facklamia, Carnobacterium, Arthrobacter, Brachybacterium, Micrococcus, Agrococcus, Leifsonia, and Flavobacterium. Most of the isolated bacteria were psychrotrophs and showed the production of one or more extracellular enzymes. Lipolytic and proteolytic activities were more prevalent among the isolates. Five isolates from the Actinobacteria phylum and one isolate from the Bacteroidetes phylum had strong pigmentation. Antibiotic susceptibility testing revealed that most of the isolates are resistant to at least one antibiotic, and seven isolates showed multi-resistance.

Species are likely to segregate their ecological niches to minimize competition for resources, but for centrally foraging predators that breed on sub-Antarctic islands in the Southern Ocean the possibility of niche segregation may be minimal. This study is the first to examine the spatial and trophic aspects of the foraging niche of sympatrically breeding macaroni and chinstrap penguins at the poorly-studied sub-Antarctic island Bouvetøya over multiple years. To measure at-sea movements and dive behavior, 90 breeding macaroni Eudyptes chrysolophus and 49 breeding chinstrap penguins Pygoscelis antarcticus were deployed with satellite transmitters and time-depth recorders over two austral summer breeding seasons, 2015 and 2018. In addition, tracked birds were sampled for blood for biogeochemical dietary analysis. Chinstrap penguins displayed large interannual variation in foraging behavior between the two years, and dove deeper, utilized larger foraging areas during late breeding stages and showed enriched values of δ15N in the first- compared to the second- year. Conversely, macaroni penguins dove to similar depths and displayed similar values of δ15N in both years. Our results suggest that potentially low krill abundances in the waters around Bouvetøya in 2015 forced the chinstrap penguins to search for alternative prey, like myctophid fishes, which resulted in increased overlap in the two species' foraging niche. Consequently, the chinstrap penguins may have faced increased interspecific competition for prey or catabolism from food shortage. Irrespective, our findings may partly explain the decreasing number of breeding chinstrap penguins at the world's most remote island, Bouvetøya.

It is established that haematological and biochemical parameters provide important data to assess the physiological condition and health status of wild birds. To undertake conservation physiology or ecophysiology work, it is therefore essential to establish baseline physiological parameters and how these parameters change with age and life history events. In this work, we determined and compared baseline haematology and serum biochemistry between adults and chicks of three Antarctic penguin species of the genus Pygoscelis: gentoo (P. papua), Adélie (P. adeliae) and chinstrap (P. antarcticus). Differences in adults among species were observed in haemoglobin and biochemical parameters such as total proteins, glucose and alkaline phosphatase activity. In addition, differences between adults and chicks in haematocrit, haemoglobin, total proteins and glucose concentration were determined. Moreover, we evaluated the electrophoretic protein profiles between adults and chicks of the genus Pygoscelis, and a conserved protein pattern was observed among species and ages in the genus. Altogether, the results suggest that biochemical and haematological differences among pygoscelids may be related to the nutritional status and energetic expenditure during breeding as well as their feeding habits and development stage. Keywords: Antarctic; haematology; physiology; Pygoscelis; penguins; serum biochemistry.

Physiological characteristics of inorganic C uptake were examined in Southern Ocean ice algae and phytoplankton assemblages. Ice algal and phytoplankton assemblages were largely dominated by diatoms and Phaeocystis antarctica, and showed a high capacity for HCO3- utilization, with direct HCO3- transport accounting for ~60% of total inorganic C uptake. Extracellular carbonic anhydrase (eCA) was detectable in all samples, but with significantly lower activity in sea ice algae. Neither HCO3- transport nor eCA activity was related to the in situ partial pressure of CO2 (pCO2) or taxonomic composition of samples. The half-saturation constant (KS) for inorganic C ranged from ~100 to 5000 µM, and showed significantly more variability among sea ice algae than phytoplankton assemblages. For the phytoplankton assemblages, there were significant positive correlations between in situ pCO2 and KS (higher C substrate affinity in low pCO2 waters), and also between KS and maximum C uptake rates (Vmax). In contrast, KS and Vmax in sea-ice algal assemblages were not correlated to each other, or to any other measured variables. The C isotope composition of particulate organic carbon(δ13C-POC) in the phytoplankton assemblages showed modest variability (range -30 to -24.6‰) and was significantly correlated to the ratio of inferred growth rates (derived from Vmax) and in situ CO2 concentrations, but not to any measured C uptake parameters. δ13C-POC in sea ice algal samples (range -25.7 to -12.9‰) was significantly heavier than in the phytoplankton assemblages, and not correlated to any other variables. Our results provide evidence for the widespread occurrence of carbon-concentrating mechanisms in Southern Ocean sea ice algae and phytoplankton assemblages. KEYWORDS: Phytoplankton · Sea ice algae · Inorganic carbon uptake · HCO3- · Carbonic anhydrase

We report the isolation and identification of bacteria that produce extracellular cold-active proteases, obtained from water samples collected near the Uruguayan Antarctic Base on King George Island, South Shetlands. The bacteria belonged to the genera Pseudomonas (growth between 4 and 30°C) and Flavobacterium (growth between 4 and 18°C). In all cases, extracellular protease production was evident when reaching the stationary phase at 18 and 4ºC, but was not detected at 30ºC. The zymogram revealed the secretion of one extracellular protease per isolate, each with different relative electrophoretic mobility. The extracellular proteases produced at 4ºC showed thermal activity and stability at 30ºC. Both activity and stability at temperature higher that 10ºC have no physiological meaning because the isolates do not experience such temperatures in the Antarctic environment; however, the possible ecological value of cold-active and -stable extracellular proteases is discussed. Keywords: Antarctic, cold-active enzymes, protease.

The venom of Antarctic octopus remains completely unstudied. Here, a preliminary investigation was conducted into the properties of posterior salivary gland (PSG) extracts from four Antarctica eledonine (Incirrata; Octopodidae) species (Adelieledone polymorpha, Megaleledone setebos, Pareledone aequipapillae, and Pareledone turqueti) collected from the coast off George V’s Land, Antarctica. Specimens were assayed for alkaline phosphatase (ALP), acetylcholinesterase (AChE), proteolytic, phospholipase A2 (PLA2), and haemolytic activities. For comparison, stomach tissue from Cirroctopus sp. (Cirrata; Cirroctopodidae) was also assayed for ALP, AChE, proteolytic and haemolytic activities. Dietary and morphological data were collected from the literature to explore the ecological importance of venom, taking an adaptive evolutionary approach. Of the incirrate species, three showed activities in all assays, while P. turqueti did not exhibit any haemolytic activity. There was evidence for cold-adaptation of ALP in all incirrates, while proteolytic activity in all except P. turqueti. Cirroctopus sp. stomach tissue extract showed ALP, AChE and some proteolytic activity. It was concluded that the AChE activity seen in the PSG extracts was possibly due to a release of household proteins, and not one of the secreted salivary toxins. Although venom undoubtedly plays an important part in prey capture and processing by Antarctica eledonines, no obvious adaptations to differences in diet or morphology were apparent from the enzymatic and haemolytic assays. However, several morphological features including enlarged PSG, small buccal mass, and small beak suggest such adaptations are present. Future studies should be conducted on several levels: Venomic, providing more detailed information on the venom compositions as well as the venom components themselves; ecological, for example application of serological or genetic methods in identifying stomach contents; and behavioural, including observations on capture of different types of prey.

Two strains of psychrotolerant Antarctic marine bacteria were isolated and characterized using biochemical and molecular techniques. Sequencing of 16S rRNA gene showed that UVvi strain belongs to the genus Arthrobacter whereas UVps strain is related to the Flexibacter-Cytophaga-Bacteroides (FCB) group. Response of the strains to solar radiation was studied during the summer of 1999 in Potter Cove, near Jubany station (South Shetland Island, Antarctica). The effect of photosynthetically available radiation (PAR, 400-700 nm), ultraviolet-A (UV-A, 320-400 nm) and ultraviolet-B radiation (UV-B, 280-320 nm) on cell viability was studied using mixed cultures in quartz bottles covered with interferential filters and exposed to solar radiation. In all experiments, four treatments were used: dark (with light screened out), PAR (with UV radiation screened out), PAR+UV-A (UV-B screened out) and PAR+UV-A+UV-B. Under the assayed conditions, PAR+UV-A and PAR+UV-A+UV-B radiation showed similar negative effects on the viability of the studied strains. However, at the end of the exposure time, mortality values in PAR+UV-A+UV-B treatments were higher than those observed under PAR+UV-A treatments. In both PAR+UV-A and PAR+UV-A+UV-B treatments we observed high levels of hydrogen peroxide compared with the dark control. The Arthrobacter UVvi strain showed significant recovery in dark conditions after exposure to the PAR+UV-A but not after the PAR+UV-A+UV-B treatment. This strain proved to be more resistant to UV radiation than the FCB group-related UVps strain. The results showed that UV radiation has a deleterious effect on these Antarctic marine bacteria and also revealed that the analysed components of the Antarctic bacterioplankton may have different responses when they are exposed to the same irradiance conditions.

Iron(III) photoreduction and the responses of phytoplankton under ultraviolet (UV) and photosynthetically available radiation (PAR) were investigated with the presence of hydroxycarboxylic acid (glucaric acid (GA), a model compound for organic acids excreted by phytoplankton). The incubation experiments were carried out on board using seawater samples collected in the location of the winter ice edge (WIE) and the spring ice edge (SIE) of the Southern Ocean. In this paper, we focus on the results of experiment in WIE. Throughout the experiments, dissolved Fe(II), major nutrients and in vivo fluorescence were monitored regularly. In addition, Chl-a, POC/PON, cell densities of phytoplankton and bacteria, bacterial production, organic peroxide, hydrogen peroxide and total CO2 were measured. The results from the WIE show that iron enrichment had a substantial effect on phytoplankton growth rate. Fe(III) addition in the presence of GA (FeGA) gave higher Fe(II) concentration and higher growth rate of phytoplankton than those in controls. Our results suggest that hydroxycarboxylic acid had a significant chemical and biological impact. The presence of GA influenced iron photochemistry and iron availability to phytoplankton. Phytoplankton growth responses to iron enrichments in incubations under UV and PAR were completely dissimilar. It seems that FeGA addition prominently changes the harmful effect of UV on the phytoplankton population. This study provides preliminary information on how the photoreduction of iron(III) and the phytoplankton growth are affected by iron enrichment in the presence of hydroxycarboxylic acid.

The role of iron and light in controlling photosynthate production and allocation in phytoplankton populations of the Atlantic sector of the Southern Ocean was investigated in April–May 1999. The 14C incorporation into five biochemical pools (glucan, amino acids, proteins, lipids and polysaccharides) was measured during iron/light perturbation experiments. The diurnal Chl a-specific rates of carbon incorporation into these pools did not change in response to iron addition, yet were decreased at 20 μmol photons m−2 s−1, an irradiance comparable with the one at 20–45 m in situ depth. This suggests that the low phytoplankton biomass encountered (0.1–0.6 μg Chl a L−1) was mainly caused by light limitation in the deep wind mixed layer (>40 m). Regional differences in Chl a-specific carbon incorporation rates were not found in spite of differences in phytoplankton species composition: at the Antarctic Polar Front, biomass was dominated by a diatom population of Fragilariopsis kerguelensis, whereas smaller cells, including chrysophytes, were relatively more abundant in the Antarctic Circumpolar Current beyond the influence of frontal systems. Because mixing was often in excess of 100 m in the latter region, diatom cells may have been unable to fulfil their characteristically high Fe demand at low average light conditions, and thus became co-limited by both resources. Using a model that describes the 14C incorporation, the consistency was shown between the dynamics in the glucan pool in the field experiments and in laboratory experiments with an Antarctic diatom, Chaetoceros brevis. The glucan respiration rate was almost twice as high during the dark phase as during the light phase, which is consistent with the role of glucan as a reserve supplying energy and carbon skeletons for continued protein synthesis during the night.

The climatic features of Antarctic waters are more extreme and constant than in the Arctic. The Antarctic has been isolated and cold longer than the Arctic. The polar ichthyofaunas differ in age, endemism, taxonomy, zoogeographic distinctiveness and physiological tolerance to environmental parameters. The Arctic is the connection between the Antarctic and the temperate-tropical systems. Paradigmatic comparisons of the pathways of adaptive evolution of fish from both poles address the oxygen-transport system and the antifreezes of northern and southern species, (i) Haemoglobin evolution has included adaptations at the biochemical, physiological and molecular levels. Within the study of the molecular bases offish cold adaptation, and taking advantage of the information on haemoglobin amino acid sequence, we analysed the evolutionary history of the ? and ? globins of Antarctic, Arctic and temperate haemoglobins as a basis for reconstructing phylogenetic relationships. In the trees, the constant physico-chemical conditions of the Antarctic waters are matched by clear grouping of globin sequences, whereas the variability typical of the Arctic ecosystem corresponds to high sequence variation, reflected by scattered intermediate positions between the Antarctic and non-Antarctic clades. (ii) Antifreeze (glyco)proteins and peptides allow polar fish to survive at sub-zero temperatures. In Antarctic Notothenioidei the antifreeze gene evolved from a trypsinogen-like serine protease gene. In the Arctic polar cod the genome contains genes which encode nearly identical proteins, but have evolved from a different genomic locus–a case of convergent evolution.

ABSTRACT: Hydrography, chlorophyll <i>a</i>, phytoplankton and zooplankton dynamics and the vertical flux of particulate organic carbon (POC) and pigments in the upper 200 m were investigated for 12 consecutive days during a drogue study conducted in the open waters of the ice-edge zone of the Lazarev Sea during the austral summer (December/January) 1994/95. Results of the study indicate that during the experiment, primary production, although variable, increased from ~300 to ~800 mgC m^-2d^-1. This increase could likely be related to development of a shallow pycnocline. Analysis of sediment trap data showed that the vertical carbon flux resulting from sedimentation and grazing activity was greatest in the upper water column (<80 m). The importance of grazers to total POC flux was highest at the beginning and the end of the investigation and accounted for up to 15% of total carbon flux. The contribution of grazers to vertical flux was negligible (<2%) during the intermediate part of the Southern Ocean Drogue study. Lower contribution of grazers to sedimentation of POC at depth can likely be related to community composition of zooplankton. Sedimentation of phytoplankton cells from the upper water column increased during the study. Here, downward POC flux resulting from sedimentation of microphytoplankton was equivalent to 15-75% of the total. Increase in sedimentation of phytoplankton during the study can be related to an increase in the average size of phytoplankton cells. Transport of POC from surface waters to deeper depths resulting from sedimentation or grazing activity was equivalent to <48% of total daily primary production, measured at 50 m, while the same value for phytoplankton flux did not exceed 27% of the total. Zooplankton density was insufficient to exert either a positive (via faecal pellets) or negative (via reducing suspended phytoplankton concentration) effect on particulate carbon sedimentation. This resulted in algal sink being the most important mechanism in downward POC flux during the onset of the phytoplankton bloom period in the Marginal Ice Zone, even in the presence of pelagic tunicates.

Zooplankton samples were collected in January 1993 off Dronning Maud Land along a transect from open waters to the marginal ice zone close to the Antarctic ice shelf. Thysanoessa macrura was caught in open waters while Calanoides acutus and Calanus propinquus were mainly sampled between ice floes in the marginal ice zone. The “ice-krill”Euphausia crystallorophias was found over the shelf directly associated with ice floes. T. macrura had a lipid content up to 36% of its dry weight with the dominant lipid class, wax ester, accounting for 45–50% of the total lipid. The predominance of 18:1 fatty alcohols is the striking characteristic of the wax esters. Small specimens of E. crystallorophias had lipid levels up to 26% of their dry weight with, unexpectedly, triacylglycerols being the dominant lipid (up to 41% of total lipid). The small levels of wax esters in these animals (3–6% of total lipid) had phytol as a major constituent. Large specimens of E. crystallorophias had up to 34% of their dry weight as lipid, with wax esters (47% of total lipid) dominated by 16:0 and 14:0 fatty alcohols as the major lipid. Calanus propinquus had lipid levels of up to 34% of their dry weight, with triacylglycerols (up to 63% of total lipid) being the dominant lipid. High levels of 22:1 (n-9) fatty acid were present in the triacylglycerols. Calanoides acutus had lipid levels up to 35% of the dry weight with wax esters accounting for up to 83% of total lipid. High levels of (n-3) polyunsaturated fatty acids were recorded with 20:5(n-3), 22:6(n-3) and 18:4(n-3) being the dominant moieties. On the basis of their lipid compositions we deduce that: (1) Calanoides acutus is the strictest herbivore among the four species studied, heavily utilizing the typical spring bloom; (2) T. macrura is essentially omnivorous, probably utilizing the less defined bloom situations found in oceanic waters; (3) E. crystallorophias is an omnivore well adapted to utilize both a bloom situation and to feed on ice algae and micro-zooplankton associated with the ice; (4) Calanus propinquus seems to be the most opportunistic feeder of the four species studied, probably grazing heavily on phytoplankton during a bloom and, during the rest of the year, feeding on whatever material is available, including particulates, flagellates and other ice-associated algae. We conclude that the different biochemical pathways generating large oil reserves of different compositions, enabling species to utilize different ecological niches, are major determinants of biodiversity in polar zooplankton.

The digestive enzyme trypsin is among the most extensively studied proteins, and its structure has been reported from a large number of organisms. This article focuses on the trypsins from vertebrates adapted to life at low temperatures. Cold-adapted organisms seem to have compensated for the reduced reaction rates at low temperatures by evolving more active and less temperature-stable enzymes. We have analyzed 27 trypsin sequences from a variety of organisms to find unique attributes for the cold-adapted trypsins, comparing trypsins from salmon, Antarctic fish, cod, and pufferfish to other vertebrate trypsins. Both the "cold" and the "warm" active trypsins have about 50 amino acids that are unique and conserved within each class. The main unique features of the cold-adapted trypsins attributable to low-temperature adaptation seem to be (1) reduced hydrophobicity and packing density of the core, mainly because of a lower (Ile + Leu)/(Ile + Leu + Val) ratio, (2) reduced stability of the C-terminal, (3) lack of one warm trypsin conserved proline residue and one proline tyrosine stacking, (4) difference in charge and flexibility of loops extending the binding pocket, and (5) different conformation of the "autolysis" loop that is likely to be involved in substrate binding.

The unique predominance of oleyl alcohols (18:1) is the striking characteristic of the lipids of the Antarctic euphausiid <i>Thysanoessa macrura</i>. The 2 isomers 18:1(n-9) and 18:1(n-7) occurred in similar proportions in the wax esters of <i>T. macrura</i> and comprised up to 80% of the total fatty alcohols. The remainder consisted mostly of the 20:1(n-9) alcohol along with small amounts of the 22:1(n-11) alcohol. No marine zooplankton species has previously been reported which produces wax esters with significant amounts of 18 carbon fatty alcohols. <i>T. macrura</i> specimens were collected in the high Antarctic Weddell Sea during autumn 1992 and summer 1993. Their lipid levels were high, about 40 to 50% of the dry mass with up to 70% of the total lipid as wax esters. The wax ester fatty acids were dominated by the saturates 14:0 and 16:0, which, along with the monounsaturate 18:1(n-9), accounted for more than 80% of the total fatty acids. Phospholipids contained high levels of (n-3) polyunsaturated fatty acids (20:5 and 22:6) typical of membrane lipids from marine zooplankton. The precise significance of the unique wax ester composition in <i>T. macrura</i> is not clear but this discovery underscores the biochemical adaptability of Antarctic zooplankton species to a constantly cold and highly seasonal polar environment.

The potential of universal calibration using capillary gas chromatography combined with atomic emission detection was evaluated for several chlorine- and sulphur-containing compounds of environmental interest. The elemental responses obtained for C, H, Cl and S were found to be affected by the molecular structure. Rapid determination of several analytes was accomplished using a single calibration graph based on one randomly selected reference compound, but 10–30% errors relative to the actual concentrations were observed in several instances. The results were significantly improved by using reference compounds with structures similar to those of the analytes of interest. the practical utility of universal calibration is illustrated by the determination of pesticides in stomach oil deposits from birds living in Antarctica.

The article discusses the new concept for surgery of necrotic ulcers using the krill enzymes. The krill peptide hydrolases represent a new and important alternative to mammalian or microbial enzymes for distinctive medical applications, such as debridemen.

Development of a comprehensive picture of the genetic population structure of the Antarctic krill (Euphausia superba) has been hampered by a lack of genetic data from two major areas of the species' distribution, the Bellingshausen Sea and the Ross Sea. Evidence from earlier studies of a discrete “Bellingshausen Sea” population was based on anomalous allele frequencies in two sample sets that were collected near the west coast of the Antarctic Peninsula rather than in the Bellingshausen Sea proper. In this paper we describe the first biochemical genetic data obtained on krill from the central Bellingshausen Sea and from the Ross Sea. Analyses of eight polymorphic loci in samples from these two areas have failed to provide any evidence of population structuring within the Pacific sector of the Southern Ocean, and have indicated that Pacific sector krill cannot be genetically discriminated from Atlantic sector krill or Indian Ocean sector krill. These findings further support the hypothesis of a single circumpolar breeding population of Antarctic krill.