caroline toney* and costantino vetriani
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Characterization of Enrichment Cultures of Nitrate-Reducing Bacteria from Deep-Sea Hydrothermal Vents. Caroline Toney [email protected] (828) 429-9476. Caroline Toney* and Costantino Vetriani. Conclusions. Introduction. - PowerPoint PPT PresentationTRANSCRIPT
Characterization of Enrichment Cultures of Nitrate-Reducing Bacteria from Deep-Sea
Hydrothermal VentsCaroline Toney* and Costantino Vetriani
Deep-sea vents are one of the last frontiers of the unexplored world. These unique environments spew forth super-heated fluid that is laden with metals. The base of life which allow other organisms to live in this ecosystem are microorganisms which undergo chemosynthesis as a means of primary production.
Introduction
Figure 2. Sampling sites of deep-sea hydrothermal vents on the East Pacific Rise
Objectives• To grow cultures from samples taken at vent sites at 9°N and 13°N. •To isolate anaerobic, nitrate-reducing bacteria using different techniquesand media.•To compare the 16S rRNA gene sequences of these organisms to the databaseusing BLAST (Basic Local Alignment Search Tool) to assess theirrelatedness to other species.
Media Carbon Source Electron Donor
Electron Acceptor
Temp (°C)
HB1 CO2 H2 NO3 50 & 75
Sulfurovum CO2 N2, thiosulfate NO3 35
Sulfurimonas CO2 H2, thiosulfate NO3 35, 50
ASW Organic N2 NO3 30
Caroline [email protected](828) 429-9476
Figure 1. Illustration of deep-sea hydrothermal vent flow. (Brock et al., 2006).
Site Media Temp (°C) Identity (%) Description
Alvinella Tubes
HB1 50 99,97 Caminibacter mediatlanticus; Nautilia profundicula, abyssi
Sulfurimonas 35, 50 86 Sulfurospirillum carboxydovorans
Alvinella sulfide
HB1 50 97, 96 Caminibacter mediatlanticus
HB1 75 98 Desulfobacterium pacificum
Crab Spa Sulfurovum 35 93;96;93 Thioprofundum lithotrophica; Rhodobacter capsulatus; Marinobacter hydrocarbonoclasticus
Sulfurimonas 35, 50 97 Shewanella haliotisAnaerobic ASW 30 100 Klebsiella pneumoniae
Bottom Seawater
Anaerobic ASW (B)
30 99 Shewanella haliotis
Samples were taken during Nautile Dive (ND) #:ND 1726 Alvinella pompejana tubesND 1727 Alvinella pompejana encrusted sulfidesND 1729 Fluid from Crab Spa; Bottom Seawater
AcknowledgementsWe want to thank the crew of the R/V L’Atlante and the crew and pilots of the DSV/Nautile for sample collection We would like to offer our thanks to the members of the Deep Sea Microbiology Lab at Rutgers University their expertise, as well.This work was supported by an NSF grant.
Table 2. Results of sequencing and BLAST.
Conclusions•The molecular characterization of the organisms cultured in the samples taken from the dives reveals that there may be new species among them.•The sequences of the 16S rRNA gene from the organisms grown show that further isolation is needed for some samples in order to have a pure culture from which further investigation of the species can be done.• Future work would include characterization of isolated organisms. Morphological observation and studies of optimal growth in temperature, salinity and pH will be executed. Metabolism will be studied through growth under different conditions (i.e. electron donors and acceptors, pressure, etc.), in organic substrates, and resistance to antibiotics will also be tested.
Table 1. Characteristics of anaerobic media used.
Alvinella pompejana. (noc.soton.ac.uk)
Crab Spa (whoi.edu)