Competition and coexistence of sulfate-reducing bacteria, acetogens and methanogens in a lab-scale anaerobic bioreactor as affected by changing substrate to sulfate ratio

The microbial population structure and function of natural anaerobic communities maintained in lab-scale continuously stirred tank reactors at different lactate to sulfate ratios and in the absence of sulfate were analyzed using an integrated approach of molecular techniques and chemical analysis. The population structure, determined by denaturing gradient gel electrophoresis and by the use of oligonucleotide probes, was linked to the functional changes in the reactors. At the influent lactate to sulfate molar ratio of 0.35 mol mol−1, i.e., electron donor limitation, lactate oxidation was mainly carried out by incompletely oxidizing sulfate-reducing bacteria, which formed 80–85% of the total bacterial population. Desulfomicrobium- and Desulfovibrio-like species were the most abundant sulfate-reducing bacteria. Acetogens and methanogenic Archaea were mostly outcompeted, although less than 2% of an acetogenic population could still be observed at this limiting concentration of lactate. In the near absence of sulfate (i.e., at very high lactate/sulfate ratio), acetogens and methanogenic Archaea were the dominant microbial communities. Acetogenic bacteria represented by Dendrosporobacter quercicolus-like species formed more than 70% of the population, while methanogenic bacteria related to uncultured Archaea comprising about 10–15% of the microbial community. At an influent lactate to sulfate molar ratio of 2 mol mol−1, i.e., under sulfate-limiting conditions, a different metabolic route was followed by the mixed anaerobic community. Apparently, lactate was fermented to acetate and propionate, while the majority of sulfidogenesis and methanogenesis were dependent on these fermentation products. This was consistent with the presence of significant levels (40–45% of total bacteria) of D. quercicolus-like heteroacetogens and a corresponding increase of propionate-oxidizing Desulfobulbus-like sulfate-reducing bacteria (20% of the total bacteria). Methanogenic Archaea accounted for 10% of the total microbial community

Geochemistry and geobiology of a present-day serpentinization site in California: The Cedars

Ultra-basic (pH 11–12) reducing (−656 to −585 mV) groundwater springs discharging from serpentinized peridotite of The Cedars, CA, were investigated for their geochemistry and geobiology. The spring waters investigated were of meteoric origin; however, geochemical modeling suggests that there were two sources of groundwater, a shallow source with sufficient contact with The Cedars’ peridotite body to be altered geochemically by serpentinization, and a deeper groundwater source that not only flows through the peridotite body but was also in contact with the marine sediments of the Franciscan Subduction Complex (FSC) below the peridotite body. We propose that the groundwater discharging from lower elevations (GPS1 and CS1) reflect the geochemistry of the deeper groundwater in contact with FSC, while groundwaters discharging from springs at higher elevations (NS1 and BSC) were a mixture of the shallow peridotite-only groundwater and the deeper groundwater that has been in contact with the FSC. Cell densities of suspended microbes within these waters were extremely low. In the NS1 and BSC spring fluids, cell densities ranged from 10^2 to 10^3 cells/ml, while suspended cells at GPS were lower than 10 cells/mL. However, glass slides incubated in the BSC and GPS1 springs for 2–3 weeks were colonized by cells with densities ranging from 10^6 to 10^7 cells/cm^2 attached to their surfaces. All of the springs were very low (⩽1 μM) in several essential elements and electron acceptors (e.g. nitrate/ammonium, sulfate, and phosphate) required for (microbial) growth, which is not uncommon at sites of continental serpentinization. Gases rich in N_2, H_2, and CH_4 were exsolving from the springs. The stable carbon isotope value (δ^(13)C_(CH4) = −68 ± 0.6‰) and the CH_4/C_(2+) (>10^3) of methane and other gaseous hydrocarbons exsolving from NS1 were typical of microbially sourced methane, whereas the isotope values and the CH_4/C_(2+) of BSC and CS1 springs were more enriched in ^(13)C and had CH_4/C_(2+) < 10^3, suggesting a mixture of microbial and non-microbial methane. The concentrations of aromatic compounds, and ethane, propane, iso- and n-butane were well described by simple physical mixing between the aromatic- and alkane-poor, shallow groundwater and the relatively aromatic, and alkane-rich groundwater that flows through both the peridotite and the FSC suggesting that these aromatic and alkane compounds originated in the deeper FSC groundwater and are not produced in the shallow peridotite-only groundwater. The aromatic compounds most probably originated from the diagenesis/degradation of organic matter in the marine sediments below the peridotite body, while the gaseous alkanes may have multiple sources including thermal degradation of the organic matter in the marine sediments below the peridotite body and possibly by abiogenic reactions occurring within the peridotite body. This geochemical study demonstrates the complexity of The Cedars, and the possible sources of hydrocarbons at continental sites of serpentinization

Mariprofundus ferrooxydans PV-1 the First Genome of a Marine Fe(II) Oxidizing Zetaproteobacterium

© The Author(s), 2011. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS One 6 (2011): e25386, doi:10.1371/journal.pone.0025386.Mariprofundus ferrooxydans PV-1 has provided the first genome of the recently discovered Zetaproteobacteria subdivision. Genome analysis reveals a complete TCA cycle, the ability to fix CO2, carbon-storage proteins and a sugar phosphotransferase system (PTS). The latter could facilitate the transport of carbohydrates across the cell membrane and possibly aid in stalk formation, a matrix composed of exopolymers and/or exopolysaccharides, which is used to store oxidized iron minerals outside the cell. Two-component signal transduction system genes, including histidine kinases, GGDEF domain genes, and response regulators containing CheY-like receivers, are abundant and widely distributed across the genome. Most of these are located in close proximity to genes required for cell division, phosphate uptake and transport, exopolymer and heavy metal secretion, flagellar biosynthesis and pilus assembly suggesting that these functions are highly regulated. Similar to many other motile, microaerophilic bacteria, genes encoding aerotaxis as well as antioxidant functionality (e.g., superoxide dismutases and peroxidases) are predicted to sense and respond to oxygen gradients, as would be required to maintain cellular redox balance in the specialized habitat where M. ferrooxydans resides. Comparative genomics with other Fe(II) oxidizing bacteria residing in freshwater and marine environments revealed similar content, synteny, and amino acid similarity of coding sequences potentially involved in Fe(II) oxidation, signal transduction and response regulation, oxygen sensation and detoxification, and heavy metal resistance. This study has provided novel insights into the molecular nature of Zetaproteobacteria.Funding has been provided by the NSF Microbial Observatories Program (KJE, DE), NSF’s Science and Technology Program, by the Gordon and Betty Moore Foundation (KJE), the College of Letters, Arts, and Sciences at the University of Southern California (KJE), and by the NASA Astrobiology Institute (KJE, DE). Advanced Light Source analyses at the Lawrence Berkeley National Lab are supported by the Office of Science, Basic Energy Sciences, Division of Materials Science of the United States Department of Energy (DE-AC02-05CH11231)

Co-existence of physiologically similar sulfate-reducing bacteria in a full-scale sulfidogenic bioreactor fed with a single organic electron donor

A combination of culture-dependent and independent methods was used to study the co-existence of different sulfate-reducing bacteria (SRB) in an upflow anaerobic sludge bed reactor treating sulfate-rich wastewater. The wastewater was fed with ethanol as an external electron donor. Twenty six strains of SRB were randomly picked and isolated from the highest serial dilution that showed growth (i.e. 108). Repetitive enterobacterial palindromic polymerase chain reaction and whole cell protein profiling revealed a low genetic diversity, with only two genotypes among the 26 strains obtained in the pure culture. The low genetic diversity suggests the absence of micro-niches within the reactor, which might be due to a low spatial and temporal micro-heterogeneity. The total 16S rDNA sequencing of two representative strains L3 and L7 indicated a close relatedness to the genus Desulfovibrio. The two strains differed in as many as five physiological traits, which might allow them to occupy distinct niches and thus co-exist within the same habitat. Whole cell hybridisation with fluorescently labeled oligonucleotide probes was performed to characterise the SRB community in the reactor. The isolated strains Desulfovibrio L3 and Desulfovibrio L7 were the most dominant SRB, representing 30–35% and 25–35%, respectively, of the total SRB community. Desulfobulbus-like bacteria contributed for 20–25%, and the Desulfobacca acetoxidans-specific probe targeted approximately 15–20% of the total SRB. The whole cell hybridisation results thus revealed a consortium of four different species of SRB that can be enriched and maintained on a single energy source in a full-scale sulfidogenic reactor

Selective enrichment of facultatively chemolithotrophic thiobacilli and related organisms in continuous culture

The current opinion is that facultatively chemo-lithotrophic thiobacilli, sometimes referred to as “versatile ” thiobacilli [ 11, often may have a signifi-cant ecological advantage over their more specialize