Incidence of plasmid and antibiotic resistance in psychrotrophic bacteria isolated from Antarctic sponges

A total of 297 bacterial strains were isolated from five Antarctic sponge species and tested by agarose gel electrophoresis for the presence of plasmid molecules. At least one kind of plasmid was carried by 69 isolates (about 23%). The disc diffusion susceptibility test was used to assay the resistance of plasmid-harbouring bacteria towards 11 antibiotics. A multiple resistance was observed for the 72% of strains, among which the 33% were resistant to only two antibiotics. Bacteria showed a high degree of resistance towards O/129 (71%), tetracycline (42%) and nalidixic acid (25%), whereas any isolate showed resistance to gentamicin. The 16S rDNA sequencing revealed that plasmid-harbouring strains were mainly affiliated to the Gammaproteobacteria (81%), whereas the other detected phylogenetic groups (i.e. Firmicutes, Alphaproteobacteria, Actinobacteria and CFB group of Bacteroidetes) were less abundant, each representing between 1% and 6% of the total isolates. The present study will contribute to the poor and fragmentary knowledge on plasmid incidence in natural microbial populations. In addition, monitoring antibiotic resistance in bacteria from remote areas, such as Antarctica, could also be a useful tool to evaluate the impact of anthropic pressure

Ricerche sulla biodiversità delle comunità batteriche in tre laghi antartici

The bacterial diversity in three Antarctic lakes located at Crater Cirque (CC), Inexpressible Island (INI) and Luther Peak (LH) in the Victoria Land was investigated by a combination of culture-dependent and culture-independent techniques. The in situ abundance of different bacterial groups was determined by fluorescent in situ hybridization (FISH), while bacterial diversity among the cultivable microflora was investigated by culturing and genetic fingerprinting. Differences in the composition of bacterial assemblages were observed among the lakes investigated, when using the two approaches reported above. Overall, the detection rate by FISH of DAPI-stained cells varied from 48.4% to 68.9% with the general bacterial probe EUB338. Bacteria hybridizing with the group-specific probe CF319a were found to be abundant in the three lakes. A total of 478 strains were isolated from R2A agar plates and grouped by restriction analysis technique; sequencing of representative 16S rDNAs was performed to elucidate the taxonomic positions of isolates. Overall, isolates were placed within five different taxa: ?-Proteobacteria (39%), Bacteroidetes (35%), ?-Proteobacteria (8%), Actinobacteria (7%) and ?-Proteobacteria (5%). Finally, the 5.8% of total isolates shared the highest degree of sequence identity with unclassified bacteria. Members of the ?-Proteobacteria predominated at INI, whereas isolates from CC and LH mainly belonged to the Bacteroidetes. Among the ?-Proteobacteria, the genus Pseudomonas was predominant, whereas Flavobacterium spp. were very common among the Bacteroidetes. Finally, several Actinobacteria were closely related to unknown Antarctic bacteria

Extracellular polymeric substances with metal adsorption capacity produced by Pseudoalteromonas sp. MER144 from Antarctic seawater

The EPS-producing Pseudoalteromonas sp. MER144 was selected among 606 isolates from Antarctic seawater due to its evident slimy appearance on agar plates. The production of EPSs was enhanced by a step-by-step approach varying the carbon source, substrate and NaCl concentrations, temperature, and pH. Optimal conditions for the EPS production resulted at temperature of 4 °C and pH 7, with addition of 2% sucrose (w/v) and 3% NaCl (w/v). EPSs produced under optimal conditions were chemically characterized, resulting in a moderate carbohydrate content (35%), uronic acids (14%), and proteins (12%). Monosaccharide composition was estimated to be Glu:Man:GluN:Ara:GluA:GalA:Gal (1:0.36:0.26:0.06:0.06:0.05:0.03), while the estimated molecular weight was about 250 kDa. The addition of sucrose in the culture medium, by stimulating the EPS production, allowed MER144 to tolerate higher concentrations of mercury and cadmium. This finding was probably dependent on the presence of uronic acids and sulfate groups, which can bind cations, in the extracted EPSs. Monitoring EPS production under optimal conditions at different concentrations of mercury and cadmium revealed that EPS amounts increased at increasing heavy metal concentrations, indicating an adaptation to the stress conditions tested

Production and biotechnological potential of extracellular polymeric substances from sponge-associated Antarctic bacteria

Four sponge-associated Antarctic bacteria (i.e. Winogradskyella spp. CAL384 and CAL396, Colwellia sp. GW185 and Shewanella sp. CAL606) were selected for the highly mucous appearance of their colonies on agar plates. The production of extracellular polymeric substances (EPSs) was enhanced by a step-by step approach varying the carbon source, substrate and NaCl concentrations, temperature and pH. The EPSs produced under optimal conditions were chemically characterized, resulting in a moderate carbohydrate content (range 15-28%), and the presence of proteins (range 3-24%) and uronic acids (range 3.2-11.9%). The chemical hydrolysis of the carbohydratic portion revealed galactose, glucose, galactosammine and mannose as principal constituents. The potential biotechnological applications of the EPSs were also investigated. The high protein content in the EPSs from Winogradskyella sp. CAL384 was probably responsible for the excellent emulsifying activity towards tested hydrocarbons, with stable emulsion index (E24) higher than those recorded for synthetic surfactants. All EPSs tested in this work improved the freeze-thaw survival ratio of isolates, thereby suggesting that they may be exploited as cryoprotection agents. The addition of a sugar in the culture medium, by stimulating the EPS production, also allowed isolates to grow in the presence of higher concentrations of mercury and cadmium. This finding was probably dependent on the presence of uronic acids and sulfate groups, which can act as ligands for cations, in the extracted EPSs

Isolation, characterization and optimization of EPSs produced by a cold-adapted Marinobacter isolate from Antarctic seawater

Marinobacter sp. W1-16 from Antarctic surface seawater was analysed for the production of extracellular polymeric substances (EPSs). Enhancement of the EPS biosynthesis was carried out by evaluating the influences of the carbon source (type and concentration), temperature, pH and salinity. EPS yields varied strongly depending on sugar substrate and temperature, while pH and salinity did not strongly affect levels of EPS production. Marinobacter sp. W1-16 produced the highest quantity of EPSs when growing at 15°C and pH 8, in the presence of 2% glucose and 3% NaCl. The EPS chemical characterization revealed a molecular weight of about 260 kDa. Colorimetric assays determined a higher quantity of carbohydrate than of proteins and uronic acids, as well as the presence of sulphate, in the extracted EPSs. The monosaccharidic composition resulted in Glc:Man:Gal:GalN:GalA:GlcA in relative molar proportions of 1:0.9:0.2:0.1:0.1:0.01. Some biotechnological potentialities (i.e. emulsifying and cryoprotective actions, and heavy metal binding properties) of the EPSs were proved, suggesting possible industrial and bioremediation applications