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University of Essex BIODEEP-WP5 Understanding of ecological relations between the microbial communities and the functioning of DHAB environments Andrea Sass , Terry McGenity, Boyd McKew Proteobacteria: mostly from interfaces some sulfur-oxidizing bacteria many related to bacteria found in deep-sea environments all grow on sea water medium, some extremely halotolerant isolates are marine bacteria derived from the oxic water column some isolates could benefit from the conditions in the interface (particularly sulfur-oxidizing bacteria) Future: investigation of an extremely halotolerant, aerobic, aerotactic strainn from Urania basin interface Bacillus related isolates: almost all derived from sediments no correlation between phylogeny and basin of origin vast majority of tested strains only halotolerant (some very halotolerant and also facultatively anaerobic) probably derived from dormant spores six strains obligately halophilic (no growth below 3.5% salt, optimum at 5-10% salt) two strains isolated anaerobically in highly saline medium Strain AS4 0.5 Series1 3% O.D. 0.4 Series2 5% 0.3 10% Series3 0.2 15% Series4 0.1 20% Series5 0 0 5 10 15 20 25 30 Time (h) 0.6 0.5 O.D. 0.4 Series1 20% 25 % Series2 0.3 30% Series3 0.2 0.1 0 0 100 200 Time (h) 300 400 Strain US13 0.5 Series1 5% Series2 10% 0.4 O.D. Series3 15% 0.3 20% Series4 22% Series5 0.2 25% Series6 0.1 27% Series7 30% Series8 0 0 100 200 300 400 Time (h) 0.4 Series1 5% 0.3 Series2 10% O.D. 15% Series3 20% Series4 0.2 22% Series5 25% Series6 27% Series7 0.1 30% Series8 0 0 10 20 Time (h) 30 other characteristics: anaerobic growth, also at high salt concentrations favours moderate temperatures and pH substrates used: amino acids, betaine, organic acids, yeast extract possibly active in situ Future: characterization of Halobacillus-related strains growing anaerobically at 20% salt Physiological properties of anerobic isolates Halanaerobiaceae : all except one (from DS) extremely halophilic distinct species from each basin, some isolated repeatedly strains derived from BI similar to isolate from another DHAB (Red Sea) CFB: moderately halophilic Clostridia: moderately halophilic Methanogens: extremely halophilic e-Proteobacterium: moderately halophilic, reduces sulfur compounds and nitrate Halorhabdus sp.: facultative anaerobe, extremely halophilic Active in situ? Methanogens, Haloanaerobiaceae and Halobacterium possibly active in situ in the brines e-Proteobacterium and CFB could thrive in the interface Clostridia probably derived from dormant spores Sequences similar to three cultivates strains were also found in clone libraries made from the basins Archaea: AN-AI3 (Halorhabdus utahensis 99%): ! 99% identity to clone DLIA-31 AN-AS7 (Methanohalophilus mahii 99%): ! 99% identity to clones ULIA-84, DLIA-189, ALIA-123, ABBA-51 Eubacteria: AN-BI4 (CFB): ! 98% identity to clone BBBB-69 (tRFLP-fragments with lengths analogous to fragments derived from AN-BI4 occur in samples from interfaces and brines) l‘Atalante sediment: extremely halophilic fermenting organisms isolated from top layers Bannock sediment: only organisms probably not active in situ isolated t-RFLP profile like that from brine no anaerobic isolates from brines l‘Atalante basin upper sediments contain bacterial population different from brine some organisms possibly active in the brines/interfaces could be cultivated vast majority of organisms inhabiting the DHAB remain uncultivated cultivation-independent approaches needed for understanding the function of DHAB environments