Microbes are known to affect ecosystems and communities as decomposers, pathogens, and mutualists. However, they also may function as classic consumers and competitors with animals if they chemically deter larger consumers from using rich food-falls such as carrion, fruits, and seeds that can represent critical windfalls to both microbes and animals. Microbes often use chemicals (i.e., antibiotics) to compete against other microbes. Thus using chemicals against larger competitors might be expected and could redirect significant energy subsidies from upper trophic levels to the detrital pathway. When we baited traps in a coastal marine ecosystem with fresh vs. microbe-laden fish carrion, fresh carrion attracted 2.6 times as many animals per trap as microbe-laden carrion. This resulted from fresh carrion being found more frequently and from attracting more animals when found. Microbe-laden carrion was four times more likely to be uncolonized by large consumers than was fresh carrion. In the lab, the most common animal found in our traps (the stone crab Menippe mercenaria) ate fresh carrion 2.4 times more frequently than microbe-laden carrion. Bacteria-removal experiments and feeding bioassays using organic extracts of microbe-laden carrion showed that bacteria produced noxious chemicals that deterred animal consumers. Thus bacteria compete with large animal scavengers by rendering carcasses chemically repugnant. Because food-fall resources such as carrion are major food subsidies in many ecosystems, chemically mediated competition between microbes and animals could be an important, common, but underappreciated interaction within many communities.

Coastal salt marshes are highly sensitive wetland ecosystems that can sustain long-term impacts from anthropogenic events such as oil spills. In this study, we examined the microbial communities of a Gulf of Mexico coastal salt marsh during and after the influx of petroleum hydrocarbons following the Deepwater Horizon oil spill. Total hydrocarbon concentrations in salt marsh sediments were highest in June and July 2010 and decreased in September 2010. Coupled PhyloChip and GeoChip microarray analyses demonstrated that the microbial community structure and function of the extant salt marsh hydrocarbon-degrading microbial populations changed significantly during the study. The relative richness and abundance of phyla containing previously described hydrocarbon-degrading bacteria ( Proteobacteria, Bacteroidetes, and Actinobacteria ) increased in hydrocarbon-contaminated sediments and then decreased once hydrocarbons were below detection. Firmicutes, however, continued to increase in relative richness and abundance after hydrocarbon concentrations were below detection. Functional genes involved in hydrocarbon degradation were enriched in hydrocarbon-contaminated sediments then declined significantly (p<0.05) once hydrocarbon concentrations decreased. A greater decrease in hydrocarbon concentrations among marsh grass sediments compared to inlet sediments (lacking marsh grass) suggests that the marsh rhizosphere microbial communities could also be contributing to hydrocarbon degradation. The results of this study provide a comprehensive view of microbial community structural and functional dynamics within perturbed salt marsh ecosystems.

A molecular phylogenetic approach was used to characterize the composition of microbial communities from two gas hydrate sedimentary systems in the Gulf of Mexico. Nucleic acids, extracted from sediments directly overlying surface-breaching gas hydrate mounds collected from a research submersible (water depth 550-575 m), were amplified with nine different 16S rDNA gene primer sets. The polymerase chain reaction primers targeted microorganisms at the domain-specific (Bacteria and Archaea) and group-specific (sulfate-reducing bacteria (SRB) and putative anaerobic methane-oxidizing (ANME) archaea) level. Amplicons were obtained with five of the nine primer sets including two of the six SRB Groups (SRB Group 5 and Group 6) and used to generate five different clone libraries. Analysis of 126 clones from the Archaea library revealed that the sediments associated with naturally occurring gas hydrate harbored a low diversity. Sequence analysis indicated the majority of archaeal clones were most closely related to Methanosarcinales, Methanomicrobiales and distinct phylogenetic lineages within the ANME groups. The most frequently recovered phylotypes in the ANME library were related to either ANME-2 or Methanomicrobiales. In contrast to the two archaeal libraries, bacterial diversity was higher with the majority of the 126 bacterial clones most closely related to uncultured clones dominated by the delta- and epsilon-Proteobacteria. Interestingly, while 82% of the clones in the SRB Group 5 library were affiliated with delta-Proteobacteria, the vast majority (83%) of clones in the SRB Group 6 library was affiliated with the Firmicutes. This is the first phylogenetic-based description of microbial communities extant in methane-rich hydrate-associated sediments from a hydrocarbon seep region in the Gulf of Mexico.

The Deepwater Horizon oil spill in the northern Gulf of Mexico represents the largest marine accidental oil spill in history. It is distinguished from past spills in that it occurred at the greatest depth (1,500 m), the amount of hydrocarbon gas (mostly methane) lost was equivalent to the mass of crude oil released, and dispersants were used for the first time in the deep sea in an attempt to remediate the spill. The spill is also unique in that it has been characterized with an unprecedented level of resolution using next-generation sequencing technologies, especially for the ubiquitous hydrocarbon-degrading microbial communities that appeared largely to consume the gases and to degrade a significant fraction of the petroleum. Results have shown an unexpectedly rapid response of deep-sea Gammaproteobacteria to oil and gas and documented a distinct succession correlated with the control of the oil flow and well shut-in. Similar successional events, also involving Gammaproteobacteria, have been observed in nearshore systems as well.

The characterization of microbial assemblages within solid gas hydrate, especially those that may be physiologically active under in situ hydrate conditions, is essential to gain a better understanding of the effects and contributions of microbial activities in Gulf of Mexico (GoM) hydrate ecosystems. In this study, the composition of the Bacteria and Archaea communities was determined by 16S rRNA phylogenetic analyses of clone libraries derived from RNA and DNA extracted from sediment-entrained hydrate (SEH) and interior hydrate (IH). The hydrate was recovered from an exposed mound located in the northern GoM continental slope with a hydrate chipper designed for use on the manned-submersible Johnson Sea Link (water depth, 550 m). Previous geochemical analyses indicated that there was increased metabolic activity in the SEH compared to the IH layer (B. N. Orcutt, A. Boetius, S. K. Lugo, I. R. Macdonald, V. A. Samarkin, and S. Joye, Chem. Geol. 205:239-251). Phylogenetic analysis of RNA-and DNA-derived clones indicated that there was greater diversity in the SEH libraries than in the IH libraries. A majority of the clones obtained from the metabolically active fraction of the microbial community were most closely related to putative sulfate-reducing bacteria and anaerobic methane-oxidizing archaea. Several novel bacterial and archaeal phylotypes for which there were no previously identified closely related cultured isolates were detected in the RNA-and DNA-derived clone libraries. This study was the first phylogenetic analysis of the metabolically active fraction of the microbial community extant in the distinct SEH and IH layers of GoM gas hydrate.Marine gas hydrates, which are ice-like crystalline solids, are composed of rigid water molecules with trapped gas molecules, primarily methane and other hydrocarbons. Gas hydrate reservoirs, which are distributed in the sediments of active and passive continental slope margins, as well as in terrestrial (i.e., permafrost) regions (38), are a proposed fossil fuel energy source (10). Additionally, the estimated global volume of submarine methane hydrates exceeds 10 16 m 3 (7, 10), highlighting the impact of hydrates on global carbon cycling, climate conditions, and seafloor stability (16,18,28,31,35). The formation of gas hydrates is dependent upon suitable gas, temperature, and pressure conditions (reviewed in reference 38). Geological and chemical conditions in the northern continental slope of the Gulf of Mexico (GoM) promote the formation of gas hydrates where seepage of hydrocarbon gases forms extensive surface-breaching mounds on the seafloor, as well as vast veinfilling hydrates in hemipelagic sediments (27).Geochemical characteristics, including gas composition and isotopic ratios of surface breaching hydrate, in the GoM have been well documented (19,33,34,38). Growth and dissolution of GoM hydrate mounds have also been observed, with changes in mound size and shape evident over a period of months (19). Such hydrate growth patterns increase fluid and solid (i.e....

Uranium contamination is an environmental concern at the Department of Energy's Field Research Center in Oak Ridge, Tennessee. In this study, we investigated whether phosphate biomineralization, or the aerobic precipitation of U(VI)-phosphate phases facilitated by the enzymatic activities of microorganisms, offers an alternative to the more extensively studied anaerobic U(VI) bioreduction. Three heterotrophic bacteria isolated from FRC soils were studied for their ability to grow and liberate phosphate in the presence of U(VI) and an organophosphate between pH 4.5 and 7.0. The objectives were to determine whether the strains hydrolyzed sufficient phosphate to precipitate uranium, to determine whether low pH might have an effect on U(VI) precipitation, and to identify the uranium solid phase formed during biomineralization. Two bacterial strains hydrolyzed sufficient organophosphate to precipitate 7395% total uranium after 120 h of incubation in simulated groundwater. The highest rates of uranium precipitation and phosphatase activity were observed between pH 5.0 and 7.0. EXAFS spectra identified the uranyl phosphate precipitate as an autunite/meta-autunite group mineral. The results of this study indicate that aerobic heterotrophic bacteria within a uranium-contaminated environment that can hydrolyze organophosphate, especially in low pH conditions, may play an important role in the bioremediation of uranium.

Aerobic heterotrophs were isolated from subsurface soil samples obtained from the U.S. Department of Energy's (DOE) Field Research Center (FRC) located at Oak Ridge, Tenn. The FRC represents a unique, extreme environment consisting of highly acidic soils with cooccurring heavy metals, radionuclides, and high nitrate concentrations. Four hundred isolates obtained from contaminated soil were assayed for heavy metal resistance, and a smaller subset was assayed for tolerance to uranium. The vast majority of the isolates were gram-positive bacteria and belonged to the high-G؉C-and low-G؉C-content genera Arthrobacter and Bacillus, respectively. Genomic DNA from a randomly chosen subset of 50 Pb-resistant (Pb r ) isolates was amplified with PCR primers specific for P IB -type ATPases (i.e., pbrA/cadA/zntA). A total of 10 pbrA/cadA/zntA loci exhibited evidence of acquisition by horizontal gene transfer. A remarkable dissemination of the horizontally acquired P IB -type ATPases was supported by unusual DNA base compositions and phylogenetic incongruence. Numerous Pb r P IB -type ATPase-positive FRC isolates belonging to the genus Arthrobacter tolerated toxic concentrations of soluble U(VI) (UO 2 2؉ ) at pH 4. These unrelated, yet synergistic, physiological traits observed in Arthrobacter isolates residing in the contaminated FRC subsurface may contribute to the survival of the organisms in such an extreme environment. This study is, to the best of our knowledge, the first study to report broad horizontal transfer of P IB -type ATPases in contaminated subsurface soils and is among the first studies to report uranium tolerance of aerobic heterotrophs obtained from the acidic subsurface at the DOE FRC.The remediation of hazardous mixed-waste sites, particularly those cocontaminated with heavy metals and radionuclides, is one of the most costly environmental challenges currently faced by the United States and other countries. Interactions between microorganisms, radionuclides, and metals that promote their precipitation and immobilization in situ are promising strategies for treatment and cleanup of the contaminated subsurface (1, 15). At mixed-waste sites where the concentrations of metal contaminants can reach toxic levels, the metal resistance of indigenous microbial populations could be critical for the success of in situ biostimulation efforts. For example, while a number of microbes can carry out reductive precipitation of radionuclides (e.g., Desulfovibrio sp., Geobacter sp., and Shewanella sp.) (28,44,63), the sensitivity of these organisms to heavy metals could possibly limit their in situ activities. Thus, the metal sensitivity of some radionuclide-reducing microbes suggests that the acquisition of metal resistance traits (e.g., P IB -type ATPases that regulate the transport of heavy metals) might be conducive to facilitating and/or enhancing microbial metabolism during subsequent biostimulation activities in metal-and radionuclide-contaminated subsurface environments.The P-type ATPases represent a chromosomally en...

In this study, the composition of the metabolically active fraction of the microbial community occurring in Gulf of Mexico marine sediments (water depth, 550 to 575 m) with overlying filamentous bacterial mats was determined. The mats were mainly composed of either orange-or white-pigmented Beggiatoa spp. Complementary 16S ribosomal DNA (crDNA) was obtained from rRNA extracted from three different sediment depths (0 to 2, 6 to 8, and 10 to 12 cm) that had been subjected to reverse transcription-PCR amplification. Domainspecific 16S PCR primers were used to construct 12 different 16S crDNA libraries containing 333 Archaea and 329 Bacteria clones. Analysis of the Archaea clones indicated that all sediment depths associated with overlying orange-and white-pigmented microbial mats were almost exclusively dominated by ANME-2 (95% of total Archaea clones), a lineage related to the methanogenic order Methanosarcinales. In contrast, bacterial diversity was considerably higher, with the dominant phylotype varying by sediment depth. An equivalent number of clones detected at 0 to 2 cm, representing a total of 93%, were related to the ␥ and ␦ classes of Proteobacteria, whereas clones related to ␦-Proteobacteria dominated the metabolically active fraction of the bacterial community occurring at 6 to 8 cm (79%) and 10 to 12 cm (85%). This is the first phylogenetics-based evaluation of the presumptive metabolically active fraction of the Bacteria and Archaea community structure investigated along a sediment depth profile in the northern Gulf of Mexico, a hydrocarbon-rich cold-seep region.