Variation of culturable bacteria along depth in the East Rongbuk ice core,Mt.Everest

Ice melt water from a 22.27 m ice core which was drilled from the East Rongbuk Glacier, Mt. Everest was incubation in two incubation ways： plate melt water directly and enrichment melt water prior plate, respectively. The abundance of cultivable bacteria ranged from 0-295 CFU mL-I to 0--1720 CFU mL-1 in two incubations with a total of 1385 isolates obtained. Comparing to direct cultivation, enrichment cultivation recovered more bacteria. Pigment-producing bacteria accounted for an average of 84.9% of total isolates. Such high percentage suggested that pigment production may be an adaptive physiological feature for the bacteria in ice core to cope with strong ultraviolet radiation on the glacier. The abundances of cultivable bacteria and pigment-producing isolates varied synchronously along depth： higher abundance in the middle and lower at the top and bottom. It indicated that the middle part of the ice core was hospitable for the microbial survival. Based on the physiological properties of the colonies, eighty-nine isolates were selected for phylogenetic analysis. Obtained 16S rRNA gene sequences fell into four groups： Firmicutes, Alpha-Proteobacteria, Gamma-Proteobacteria, and Actinobacteria, with the Firmicutes being dominant. Microbial compositions derived from direct and enrichment cultivations were not overlapped. We suggest that it is a better way to explorethe culturable microbial diversity in ice core by combining the approaches of both direct and enrichment cultivation.

Antimicrobial activities of novel cultivable bacteria isolated from marine sponge Tedania anhelans

Marine sponge Tedania anhelans distributes throughout the intertidal zone of Fujian, southeastern China, and is a potential source of natural bioactive products. The sponge harbors a large number of bacterial groups that have been identified using various techniques, including fluorescent in situ hybridization （FISH）. Fractionation of dissociated sponge allowed isolation of 25 bacterial species. Based on 16S rRNA gene sequencing, phylogenetic analysis attributed most of these eubacteria to cL-Proteobacteria, y-Proteobacteria, Cytophaga/Flavobacterium/Bacteroidetes （CFB group）, and the family Bacillaceae of Gram-positive bacteria. In sequence similarity, five putatively novel species were identified with less than 98% similarity to other strains in the NCBI database. Tests for antimicrobial activities were performed against Gram-positive bacteria, Gram-negative bacteria, fungi, antitumor indicators Escherichia coli 343/591 （with DNA repair deficiency）, regular E. coli 343/636 （with different DNA repair capacity）, and 10 bacterial isolates exhibited inhibitory bioactivities. Among these strains, three isolates were detected involving function gene NRPS-A domains, which were most closely related to the amino acid sequences of linear gramicidin synthetase and pyoverdine synthetase. These results contribute to our knowledge of the microbes associated with marine sponges and further reveal novel bacterial resources for the screening of bioactive marine natural products.

Phylogenetic analysis of cultivable bacteria isolated from Arctic sea-ice

Phylogenetic analysis based on 16S rDNA of 8 strains of cultivable bacteria isolated from Arctic sea-ice was studied.The results showed that strain BJ1 belonged to genus Planococcus,which was a genus of low mole percent G+C gram-positive bacteria;strain BJ6 belonged to genus Burkholderia of β-proteobacteria and the rest 6 strain all belonged to γ-proteobacteria,of which strain BJ8 was a species of Pseudoalteromonas,strain BJ2-BJ5 and BJ7 were members of genus Psychrobacter.Phylogenetic analysis also indicated that bacteria of genus Psychrobacter of the isolates formed a relatively independent phylogenetic cluster in comparison with other bacteria belonged to genus Psychrobacter.

Analysis of bacterial community in bulking sludge using culture-dependent and -independent approaches

The bacterial community of a bulking sludge from a municipal wastewater treatment plant with anoxic-anaerobic-oxic process was investigated by combination of cultivation and 16S rRNA gene clone library analysis for understanding the causes of bulking.A total of 28 species were obtained from 63 isolates collected from six culture media.The most cultivable species belonged to γ-Proteobacteria including Klebsiella sp.,Pseudomonas sp.,Aeromonas sp.and Acinetobacter sp.Further analysis of these strains by repetitive sequence based on polymerase chain reaction （rep-PCR） technology showed that rep-PCR yielded discriminatory banding patterns within the same genus using REP and BOX primer sets.While the culture-independent assessment revealed that β-Proteobacteria was the dominant group in the bulking sample.Sequence analysis revealed that the highest proportion （14.7%） of operational taxonomic units was 98% similar to Candidatus Accumulibacter phosphatis,which is used to remove phosphorous from wastewater.Our results indicated that combining different approaches can produce complementary information,thus generate a more accurate view of microbial community in bulking sludge.