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Global climate change is significantly affecting marine life off the northern tip of the Antarctic Peninsula, but little is known about microbial ecology in this area. The main goal of this study was to investigate the bacterioplankton community structure in surface waters using pyrosequencing and to determine factors influencing this community. Pelagibacterales and Rhodobacterales (Alphaproteobacteria), Oceanospirillales and Alteromonadales (Gammaproteobacteria), and Flavobacteriales (Bacteroidetes) were the core taxa in our samples, and the five most relatively abundant genera were Pelagibacter, Polaribacter, Octadecabacter, group HTCC2207 and Sulfitobacter. Although nutrients and chlorophyll a (chl a) contributed more to bacterioplankton community structure than water masses or depth, only 30.39% of the variance could be explained by the investigated environmental factors, as revealed by RDA and pRDA. No significant difference with respect to nutrients and chl a was observed among water masses or depth, as indicated by ANOVA. Furthermore, significant correlations among the dominant bacterial genera were more common than correlations between dominant genera and environmental factors, as revealed by Spearman analysis. We conclude that nutrients and chl a become homogeneous and that interpopulation interactions may have a central role in influencing the bacterial community structure in surface waters off the northern tip of the Antarctic Peninsula during the summer.

A previously uncultured cyanobacterium, strain KNUA009, was axenically isolated from a meltwater stream on Barton Peninsula, King George Island, South Shetland Islands, Antarctica. Molecular evidences showed that the isolate belongs to groups of globally distributed cryosphere cyanobacterial clones and this new isolate represents the first laboratory culture to be assigned to these groups. Strain KNUA009 was able to thrive at low temperatures ranging between 5°C and 20°C, but did not survive at temperatures of 25°C and above. As the isolate morphologically resembled Oscillatoria species, it is suggested that this cyanobacterium may represent a new species clade with cold resistance within the genus Oscillatoria. Keywords: Barton Peninsular; cryosphere cyanobacteria; King George Island; uncultured Oscillatoria species.

Affiliations of the dominant culturable bacteria isolated from Potter Cove, South Shetland Islands, Antarctica, were investigated together with their production of cold-active hydrolytic enzymes. A total of 189 aerobic heterotrophic bacterial isolates were obtained at 4°C and sorted into 63 phylotypes based on their amplified ribosomal DNA restriction analysis profiles. The sequencing of the 16S rRNA genes of representatives from each phylotype showed that the isolates belong to the phyla Proteobacteria (classes Alpha- and Gamma-proteobacteria), Bacteroidetes (class Flavobacteria), Actinobacteria (class Actinobacteria) and Firmicutes (class Bacilli). The predominant culturable group in the site studied belongs to the class Gammaproteobacteria, with 65 isolates affiliated to the genus Pseudoalteromonas and 58 to Psychrobacter. Among the 189 isolates screened, producers of amylases (9.5%), pectinases (22.8%), cellulases (14.8%), CM-cellulases (25.4%), xylanases (20.1%) and proteases (44.4%) were detected. More than 25% of the isolates produced at least one extracellular enzyme, with some of them producing up to six of the tested extracellular enzymatic activities. These results suggest that a high culturable bacterial diversity is present in Potter Cove and that this place represents a promising source of biomolecules. Keywords: Microbial enzymes; Antarctic bacteria; marine bacteria; cold enzymes; psychrophiles.

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.

There is a growing concern about the ability to produce enough nutritious food to feed the global human population in this century. Environmental conflicts and a limited freshwater supply constrain further developments in agriculture; global fisheries have levelled off, and aquaculture may have to play a more prominent role in supplying human food. Freshwater is important, but it is also a major challenge to cultivate the oceans in an environmentally, economically and energy-friendly way. To support this, a long-term vision must be to derive new sources of feed, primarily taken from outside the human food chain, and to move carnivore production to a lower trophic level. The main aim of this paper is to speculate on how feed supplies can be produced for an expanding aquaculture industry by and beyond 2050 and to establish a roadmap of the actions needed to achieve this. Resources from agriculture, fish meal and fish oil are the major components of pellet fish feeds. All cultured animals take advantage of a certain fraction of fish meal in the feed, and marine carnivores depend on a supply of marine lipids containing highly unsaturated fatty acids (HUFA, with ≥3 double bonds and ≥20 carbon chain length) in the feed. The availability of HUFA is likely the main constraint for developing carnivore aquaculture in the next decades. The availability of fish meal and oil will decrease, and the competition for plant products will increase. New harvested resources are herbivore zooplankton, such as Antarctic krill and red feed, and new produced resources are macroalgae, transgenic higher HUFA-producing plants and bacterial biomass. These products are to a limited extent components of the human food chain, and all these resources will help to move cultured carnivores to lower trophic levels and can thereby increase the production capacity and the sustainability of the production. Mariculture can only become as successful as agriculture in the coming century if carnivores can be produced at around Trophic Level 2, based mainly on plant resources. There is little potential for increasing the traditional fish meal food chain in aquaculture. KEYWORDS: Global aquaculture · Mariculture · Feed resources · Marine lipids · HUFA · Trophic level

Antarctic bacteria producing extracellular lipolytic enzymes with activity at low temperature were isolated, and the most promising strain, named G, was identified as a Psychrobacter species based on 16S rDNA sequence alignment. The genomic DNA of this bacterium was used to construct its plasmid genomic library into pUC118 plasmid vectors, and to screen the cold-active lipolytic enzyme genes. Two genes encoding for cold-active lipolytic enzymes, Lip-1452 (with an open reading frame of 1452 bp in length) and Lip-948 (with an open reading frame of 948 bp in length), were screened. The primary structure of the two lipases deduced from the nucleotide sequence showed a consensus pentapeptide containing the active serine (Lip-1452, GDSAG, and Lip-948, GNSMG) and a conserved His-Gly dipeptide in the N-terminal part of the enzyme. Protein sequence alignment and conserved regions analysis indicated that the two lipases probably belonged to family IV and family V of the bacterial lipolytic enzymes, respectively. The upstream and downstream sequences of the two lipolytic lipases were also obtained. The two lipase genes were cloned into the expression vector pCold III and integrated into Escherichia coli BL21 (DE3). The functional expression of both lipase genes by E. coli BL21 (DE3) cells was observed as the formation of clear haloes around colonies on a 1% (vol/vol) tributyrin plate upon induction with isopropyl-b-Dthiogalactopyranoside at 5°C. A lipase activity assay showed that the specific activity of the pCold III+Lip-948 expression system was up to 3.7 U ml-1, whereas that of pCold III+Lip-1452 was very low.