Microbial diversity and methanogenic activity of Antrim Shale formation waters from recently fractured wells.

The Antrim Shale in the Michigan Basin is one of the most productive shale gas formations in the U.S., but optimal resource recovery strategies must rely on a thorough understanding of the complex biogeochemical, microbial, and physical interdependencies in this and similar systems. We used Illumina MiSeq 16S rDNA sequencing to analyze the diversity and relative abundance of prokaryotic communities present in Antrim shale formation water of three closely spaced recently fractured gas-producing wells. In addition, the well waters were incubated with a suite of fermentative and methanogenic substrates in an effort to stimulate microbial methane generation. The three wells exhibited substantial differences in their community structure that may arise from their different drilling and fracturing histories. Bacterial sequences greatly outnumbered those of archaea and shared highest similarity to previously described cultures of mesophiles and moderate halophiles within the Firmicutes, Bacteroidetes, and δ- and ε-Proteobacteria. The majority of archaeal sequences shared highest sequence similarity to uncultured euryarchaeotal environmental clones. Some sequences closely related to cultured methylotrophic and hydrogenotrophic methanogens were also present in the initial well water. Incubation with methanol and trimethylamine stimulated methylotrophic methanogens and resulted in the largest increase in methane production in the formation waters, while fermentation triggered by the addition of yeast extract and formate indirectly stimulated hydrogenotrophic methanogens. The addition of sterile powdered shale as a complex natural substrate stimulated the rate of methane production without affecting total methane yields. Depletion of methane indicative of anaerobic methane oxidation (AMO) was observed over the course of incubation with some substrates. This process could constitute a substantial loss of methane in the shale formation.

Novel strains isolated from a coastal aquifer suggest a predatory role for flavobacteria.

Three newly isolated strains of flavobacteria from coastal aquifer sediments have been found to be predatory, lysing a range of live and pasteurized microbial prey. The three strains have been classified on the basis of 16S rRNA gene phylogeny as belonging to the recently described Olleya (strains VCSA23 and VCSM12) and Tenacibaculum (strain VCSA14A) genera. Two of the closest cultured relatives to the strain VCSA14A, Tenacibaculum discolor and Tenacibaculum gallaicum, were also found to be bacteriolytic. These five predatory strains exhibit gliding motility and have been observed to lyse prey cells after surrounding them with social swarms, similar to known predatory bacteria such as myxobacteria and members of the genus Lysobacter. Flavobacteria are often numerically significant in a wide variety of freshwater and marine environments, particularly in association with particles, and are thought to be involved in the degradation of biopolymeric substances. If predatory capability is widespread among flavobacteria, they may be a previously unrecognized source of 'top-down' bacterial mortality with an influence on the composition and activity of surrounding microbial communities.

Abundance and diversity of microbial life in ocean crust.

Oceanic lithosphere exposed at the sea floor undergoes seawater-rock alteration reactions involving the oxidation and hydration of glassy basalt. Basalt alteration reactions are theoretically capable of supplying sufficient energy for chemolithoautotrophic growth. Such reactions have been shown to generate microbial biomass in the laboratory, but field-based support for the existence of microbes that are supported by basalt alteration is lacking. Here, using quantitative polymerase chain reaction, in situ hybridization and microscopy, we demonstrate that prokaryotic cell abundances on seafloor-exposed basalts are 3-4 orders of magnitude greater than in overlying deep sea water. Phylogenetic analyses of basaltic lavas from the East Pacific Rise (9 degrees N) and around Hawaii reveal that the basalt-hosted biosphere harbours high bacterial community richness and that community membership is shared between these sites. We hypothesize that alteration reactions fuel chemolithoautotrophic microorganisms, which constitute a trophic base of the basalt habitat, with important implications for deep-sea carbon cycling and chemical exchange between basalt and sea water.