Response of bacterial isolates from Antarctic shallow sediments towards heavy metals, antibiotics and polychlorinated biphenyls.

The response of bacterial isolates from Antarctic sediments to polychlorinated biphenyls (Aroclor 1242 mixture), heavy metal salts (cadmium, copper, mercury and zinc) and antibiotics (ampicillin, chloramphenicol, kanamycin and streptomycin) was investigated. Overall, the ability to growth in the presence of Aroclor 1242 as a sole carbon source was observed for 22 isolates that mainly belonged to Psychrobacter spp. Tolerance to the heavy metals assayed in this study was in the order of Cd > Cu > Zn > Hg and appeared to be strictly related to the metal concentrations, as determined during previous chemical surveys in the same area. With regards to antibiotic assays, the response of the isolates to the tested antibiotics ranged from complete resistance to total susceptibility. In particular, resistances to ampicillin and chloramphenicol were very pronounced in the majority of isolates. Our isolates differently responded to the presence of toxic compounds primarily based on their phylogenetic affiliation and secondarily at strain level. Moreover, the high incidence of resistance either to metal or antibiotics, in addition to the capability to grow on PCBs, confirm that bacteria are able to cope and/or adapt to the occurrence pollutants even in low human-impacted environments.

Marine bacterioplankton diversity and community composition in an antarctic coastal environment.

The bacterial community inhabiting the water column at Terra Nova Bay (Ross Sea, Antarctica) was examined by the fluorescent in situ hybridization (FISH) technique and the genotypic and phenotypic characterization of 606 bacterial isolates. Overall, the FISH analysis revealed a bacterioplankton composition that was typical of Antarctic marine environments with the Cytophaga/Flavobacter (CF) group of Bacteroidetes that was equally dominant with the Actinobacteria and Gammaproteobacteria. As sampling was performed during the decay of sea-ice, it is plausible to assume the origin of Bacteroidetes from the sea-ice compartment where they probably thrive in high concentration of DOM which is efficiently remineralized to inorganic nutrients. This finding was supported by the isolation of Gelidibacter, Polaribacter, and Psychroflexus members (generally well represented in Antarctic sea-ice) which showed the ability to hydrolyze macromolecules, probably through the production of extracellular enzymes. A consistently pronounced abundance of the Gammaproteobacteria (67.8%) was also detected within the cultivable fraction. Altogether, the genera Psychromonas and Pseudoalteromonas accounted for 65.4% of total isolates and were ubiquitous, thus suggesting that they may play a key role within the analyzed bacterioplankton community. In particular, Pseudoalteromonas isolates possessed nitrate reductase and were able to hydrolyze substrates for protease, esterase, and ß-galactosidase, thus indicating their involvement in the carbon and nitrogen cycling. Finally, the obtained results highlight the ability of the Actinobacteria to survive and proliferate in the Terra Nova Bay seawater as they generally showed a wide range of salt tolerance and appeared to be particularly competitive with strictly marine bacteria by better utilizing supplied carbon sources.

Antagonistic interactions between psychrotrophic cultivable bacteria isolated from Antarctic sponges: a preliminary analysis.

The present work was aimed at studying antagonistic interactions existing among cultivable bacteria associated with the Antarctic sponges Anoxycalyx joubini and Lissodendoryx nobilis. Overall, bacterial isolates were affiliated with the alpha- and gamma-Proteobacteria (17.3 and 65.3%, respectively), the CFB group of Bacteroidetes (10.7%) and the Actinobacteria (6.7%) by 16S rDNA sequencing. The two sponges harbored microorganisms belonging to different species/genera and previously retrieved from polar marine environments. Antagonistic interactions, assayed by the cross-streak method and statistically analyzed using the "network theory" approach, were checked among isolates associated with the same sponge as well as between isolates retrieved from the two sponge species ("cross-niche inhibition"). Results suggest that antagonism could play a significant role in shaping bacterial communities within sponge tissues. Data from this study confirm previous observations on the antibacterial activity of Antarctic microorganisms and represent a baseline for further investigation of both the ecological role and biotechnological exploitation of Antarctic sponge-associated bacteria.

Effects of a Simulated Acute Oil Spillage on Bacterial Communities from Arctic and Antarctic Marine Sediments.

BACKGROUND: The bacterial community responses to oil spill events are key elements to predict the fate of hydrocarbon pollution in receiving aquatic environments. In polar systems, cold temperatures and low irradiance levels can limit the effectiveness of contamination removal processes. In this study, the effects of a simulated acute oil spillage on bacterial communities from polar sediments were investigated, by assessing the role of hydrocarbon mixture, incubation time and source bacterial community in selecting oil-degrading bacterial phylotypes. METHODS: The bacterial hydrocarbon degradation was evaluated by gas chromatography. Flow cytometric and fingerprinting profiles were used to assess the bacterial community dynamics over the experimental incubation time. RESULTS: Direct responses to the simulated oil spill event were found from both Arctic and Antarctic settings, with recurrent bacterial community traits and diversity profiles, especially in crude oil enrichment. Along with the dominance of Pseudomonas spp., members of the well-known hydrocarbon degraders Granulosicoccus spp. and Cycloclasticus spp. were retrieved from both sediments. CONCLUSIONS: Our findings indicated that polar bacterial populations are able to respond to the detrimental effects of simulated hydrocarbon pollution, by developing into a more specialized active oil degrading community.

Bacterium-bacterium inhibitory interactions among psychrotrophic bacteria isolated from Antarctic seawater (Terra Nova Bay, Ross Sea).

One hundred and forty bacteria isolated from Antarctic seawater samples were examined for their ability to inhibit the growth of indigenous isolates and their sensitivity to antibacterial activity expressed by one another. On the basis of 16S rRNA gene sequencing and analysis, bacterial isolates were assigned to five phylogenetically different taxa, Actinobacteria, alpha and gamma subclasses of Proteobacteria, Bacillaceae, and Bacteroidetes. Twenty-one isolates (15%), predominantly Actinobacteria, exhibited antagonistic properties against marine bacteria of Antarctic origin. Members of Bacteroidetes and Firmicutes did not show any inhibitory activity. Differences were observed among inhibition patterns of single isolates, suggesting that their activity was more likely strain-specific rather than dependent on phylogenetic affiliation. A novel analysis based on network theory confirmed these results, showing that the structure of this population is probably robust to perturbations, but also that it depends strongly on the most active strains. The determination of plasmid incidence in the bacterial strains investigated revealed that there was no correlation between their presence and the antagonistic activity. The data presented here provide evidence for the antagonistic interactions within bacterial strains inhabiting Antarctic seawater and suggest the potential exploitation of Antarctic bacteria as a novel source of antibiotics.

Biodegradative potential and characterization of psychrotolerant polychlorinated biphenyl-degrading marine bacteria isolated from a coastal station in the Terra Nova Bay (Ross Sea, Antarctica).

Antarctic marine bacteria were screened for their ability to degrade polychlorinated biphenyls (PCB) as the sole carbon and energy source at both 4 degrees C and 15 degrees C. PCB-degrading isolates (7.1%) were identified by sequencing their 16S rDNA as Pseudoalteromonas, Psychrobacter and Arthrobacter members. One representative isolate per genera was selected for evaluating the biodegradative potential under laboratory scale and phenotypically characterized. Removal of individual PCB congeners was between 35.6% and 79.8% at 4 degrees C and between 0.4% and 82.8% at 15 degrees C. Differences in the removal patterns of PCB congeners were observed in relation to the phylogenetic affiliation: Arthrobacter isolate showed similar biodegradation efficiencies when growing at 4 degrees C and 15 degrees C, while Pseudoalteromonas better degraded PCBs at 15 degrees C. No biodegradation was detected for Psychrobacter isolate at 4 degrees C. Results obtained highlight the occurrence of PCB-degrading bacteria in Antarctic seawater and suggest the potential exploitation of autochthonous bacteria for PCB bioremediation in cold marine environments.

Biodiversity of cultivable psychrotrophic marine bacteria isolated from Terra Nova Bay (Ross Sea, Antarctica).

A set of 146 Antarctic marine isolates from the Ross Sea was characterized by a combination of molecular techniques in order to determine the degree of inter- and intraspecific variability. Isolates were analyzed by amplified rDNA restriction analysis (ARDRA) using the tetrameric enzyme AluI, resulting in 52 different groups, corresponding to at least 52 different bacterial species, indicating a high degree of interspecific variability. The phylogenetic position of bacteria belonging to some ARDRA groups was obtained by sequencing of 16S rDNA. Random amplified polymorphic DNA (RAPD) analysis, carried out on the largest ARDRA groups, revealed a high intraspecific genetic variability, too. The analysis of plasmid content revealed the existence of horizontal gene transfer between strains belonging to the same and to different species. A comparison of the whole body of morphological, physiological and biochemical data was finally carried out.

The biodegradation efficiency on diesel oil by two psychrotrophic Antarctic marine bacteria during a two-month-long experiment.

Two psychrotrophic bacterial strains isolated from Antarctic seawaters were investigated for their capability to degrade commercial diesel oil. The efficiency of hydrocarbon utilization was studied at 4 and 20 degrees C over a period of two-months. Strains were cultured in a mineral liquid medium supplemented with diesel oil as the sole source of carbon and energy. The viable counts for the bacterial abundance estimation and the culture extractions for the subsequent gas-chromatographic analysis were carried out simultaneously. The biodegradation efficiency was higher at 20 degrees C than at 4 degrees C for both strains and the decrease in hydrocarbon concentrations reached more than 85% after 60 days of incubation at 20 degrees C. Our results suggest the possible exploitation of these two bacterial strains in future biotechnological processes, directly as field-released micro-organisms both in cold and temperate contaminated marine environments.

Predominance of Flavobacterium, Pseudomonas, and Polaromonas within the prokaryotic community of freshwater shallow lakes in the northern Victoria Land, East Antarctica.

A polyphasic approach that included PCR-dependent and PCR-independent molecular techniques was applied to analyze the prokaryotic community in surface waters of shallow Antarctic lakes. The in situ abundance of different bacterial groups was determined by the fluorescence in situ hybridization, whereas bacterial diversity was investigated by 16S rRNA gene sequencing of bacterial clones and isolates. The different approaches allowed the identification of the significant microbial components of the lake bacterioplanktonic communities, indicating a predominance of Flavobacterium, Pseudomonas, and Polaromonas (up to about 56% of total sequences). These genera also appear to be important in freshwater systems elsewhere in the world. Interestingly, closest blast matches to our sequences were predominantly from polar lakes and ponds, in addition to streams and glaciers, suggesting a bipolar distribution of freshwater lake bacterioplankton. Bacteria that are more traditionally associated with the marine environment were also detected, thus indicating an external input by atmospheric deposition and/or seabird excreta. Finally, a slightly different microbial community occurred in the lake at Inexpressible Island that was characterized by low N  :  P ratio and very high conductivity value, reinforcing the idea that physicochemical and trophic status may affect the structure and composition of the bacterioplankton assemblages in Antarctic lakes.

Predation impact of ciliated and flagellated protozoa during a summer bloom of brown sulfur bacteria in a meromictic coastal lake.

Anaerobic phagotrophic protozoa may play an important role in the carbon flux of chemically stratified environments, especially when phototrophic sulfur bacteria account for a high proportion of the primary production. To test this assumption, we investigated the vertical and temporal distribution of microbial heterotrophs and of autotrophic picoplankton throughout the water column of the meromictic coastal lake Faro (Sicily, Italy), in the summer of 2004, coinciding with a bloom of brown-colored green sulfur bacteria. We also assessed the grazing impact of ciliated and flagellated protozoa within the sulfur bacteria plate using a modification of the fluorescently labeled bacteria uptake approach, attempting to minimize the biases intrinsic to the technique and to preserve the in situ anoxic conditions. Significant correlations were observed between ciliate biomass and bacteriochlorophyll e concentration, and between heterotrophic nanoflagellate biomass and chlorophyll a concentration in the water column. The major predators of anaerobic picoplankton were pleuronematine ciliates and cryptomonad flagellates, with clearances of 26.6 and 9.5 nL per cell h(-1), respectively, and a cumulative impact on the picoplankton gross growth rate ranging between 36% and 72%. We concluded that protozoan grazing channels a large proportion of anaerobic picoplankton production to higher trophic levels without restraining photosynthetic bacteria productivity.

Characterization of Antarctic psychrotrophic bacteria with antibacterial activities against terrestrial microorganisms.

Five-hundreds and eighty bacterial strains, isolated from various Antarctic marine sources and locations, were screened for antimicrobial activity against terrestrial microorganisms. Twenty-two Antarctic isolates (3.8%), mainly retrieved from the water column at Terra Nova Bay (Ross Sea), expressed antagonistic activity against one to three indicator organisms. Escherichia coli and Proteus mirabilis resulted as the more susceptible, followed by Micrococcus luteus and Bacillus subtilis. None of the isolates inhibited the growth of Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella enterica and the eukaryotic fungus Candida albicans. Active Antarctic isolates, identified by 16S rDNA sequencing and phenotypically characterized by classical methods, were phylogenetically affiliated to the Actinobacteria (16 strains) and the gamma-Proteobacteria (6 strains). Inhibition patterns, as well as phenotypic characteristics, highly vary for different isolates, even though they were affiliated to the same genus or closely related to the identical microorganism retrieved from the database, suggesting that these features were more likely strain-rather than species-specific.Results obtained from the present study confirm previous observations and highlight the potentiality of Antarctic marine bacteria as novel source of antibacterial substances.

Crude oil-induced structural shift of coastal bacterial communities of rod bay (Terra Nova Bay, Ross Sea, Antarctica) and characterization of cultured cold-adapted hydrocarbonoclastic bacteria.

For preliminary screening of human impact on Antarctic coasts, the compositions of microbial communities were analyzed in seawater at two sites located in the Terra Nova Bay of Antarctica (Ross Sea) by a combination of 16S rRNA gene sequencing and culture techniques. The bacterial community in the sample from the Rod Bay site, located at the proximity to the Italian Station, was characterized by a high abundance of 16S rRNA gene sequences belonging to the microflora typically found in soil and freshwater environments. In contrast, the seawater sample from the Adelie Cove station, a pristine reference site, contained 16S rRNA gene sequences typically found in marine areas affected by algal blooms and sea ice decay. The addition of crude oil to the Rod Bay seawater sample rapidly induced a shift in the composition of the bacterial community with appearance of novel taxonomic groups and a dramatic increase in the relative abundance of gamma-Proteobacteria sequences, whereas no significant changes were detected in the bacterial community of the Adelie Cove sample under the same conditions. Bacteria-exhibiting features with potential interest for industrial and environmental applications were isolated from the Rod Bay oil-enriched sample. In particular, hydrocarbon-degrading, cold-adapted bacteria were selectively enriched, isolated and screened for their ability to synthesize polyunsaturated fatty acids. Twenty two bacterial strains were isolated from the oil enrichment culture and identified. Eighteen isolates were found to be members of gamma-Proteobacteria, while the remainder were representatives of alpha-Proteobacteria, CFB and high G + C divisions.