Roles of Organohalide-Respiring Dehalococcoidia in Carbon Cycling

ABSTRACT The class Dehalococcoidia within the Chloroflexi phylum comprises the obligate organohalide-respiring genera Dehalococcoides, Dehalogenimonas, and “Candidatus Dehalobium.” Knowledge of the unique ecophysiology and biochemistry of Dehalococcoidia has been largely derived from studies with enrichment cultures and isolates from sites impacted with chlorinated pollutants; however, culture-independent surveys found Dehalococcoidia sequences in marine, freshwater, and terrestrial biomes considered to be pristine (i.e., not impacted with organohalogens of anthropogenic origin). The broad environmental distribution of Dehalococcoidia, as well as other organohalide-respiring bacteria, supports the concept of active halogen cycling and the natural formation of organohalogens in various ecosystems. Dechlorination reduces recalcitrance and renders organics susceptible to metabolic oxidation by diverse microbial taxa. During reductive dechlorination, hydrogenotrophic organohalide-respiring bacteria, in particular Dehalococcoidia, can consume hydrogen to low consumption threshold concentrations (<0.3 nM) and enable syntrophic oxidation processes. These functional attributes and the broad distribution imply that Dehalococcoidia play relevant roles in carbon cycling in anoxic ecosystems.

Acetate turnover and methanogenic pathways in Amazonian lake sediments

Lake sediments in Amazonia are a significant source of CH4, a potential greenhouse gas. Previous studies of sediments using 13C analysis found that the contribution of hydrogenotrophic versus acetoclastic methanogenesis to CH4 production was relatively high. Here, we determined the methanogenic pathway in the same sediments (n=6) by applying 14Cbicarbonate or 2-14Cacetate and confirmed the high relative contribution (50 %–80 %) of hydrogenotrophic methanogenesis. The respiratory index (RI) of 2-14Cacetate, which is 14CO2 relative to 14CH4+14CO2, divided the sediments into two categories, i.e., those with an RI < 0.2 consistent with the operation of acetoclastic methanogenesis and those with an RI > 0.4 showing that a large percentage of the acetate-methyl was oxidized to CO2 rather than reduced to CH4. Hence, part of the acetate was probably converted to CO2 plus H2 via syntrophic oxidation, thus enhancing hydrogenotrophic methanogenesis. This happened despite the presence of potentially acetoclastic Methanosaetaceae in all the sediments. Alternatively, acetate may have been oxidized with a constituent of the sediment organic matter (humic acid) serving as oxidant. Indeed, apparent acetate turnover rates were larger than CH4 production rates except in those sediments with a R<0.2. Our study demonstrates that CH4 production in Amazonian lake sediments was not simply caused by a combination of hydrogenotrophic and acetoclastic methanogenesis but probably involved additional acetate turnover.

Acetate turnover and methanogenic pathways in Amazonian lake sediments

Lake sediments in Amazonia are a significant source of CH4, a potential greenhouse gas. Previous studies of sediments using 13C analysis found that the contribution of hydrogenotrophic versus aceticlastic methanogenesis to CH4 production was relatively high. Here, we determined the methanogenic pathway in the same sediments (n = 6) by applying [14C]bicarbonate or [2-14C]acetate, and confirmed the high relative contribution (50–80 %) of hydrogenotrophic methanogenesis. The respiratory index (RI) of [2-14C]acetate, which is 14CO2 relative to 14CH4 &amp;plus; 14CO2, divided the sediments into two categories, i.e., those with an RI 0.4 showing that a large percentage of the acetate-methyl was oxidized to CO2 rather than reduced to CH4. Hence, part of the acetate was probably converted to CO2 plus H2 via syntrophic oxidation, thus enhancing hydrogenotrophic methanogenesis. This happened despite the presence of potentially aceticlastic Methanosaetaceae in all the sediments. Alternatively, acetate may have been oxidized with a constituent of the sediment organic matter (humic acid) serving as oxidant. Indeed, apparent acetate turnover rates were larger than CH4 production rates except in those sediments with a R

Syntrophic Oxidation of Propionate in Rice Field Soil at 15 and 30°C under Methanogenic Conditions

ABSTRACTPropionate is one of the major intermediary products in the anaerobic decomposition of organic matter in wetlands and paddy fields. Under methanogenic conditions, propionate is decomposed through syntrophic interaction between proton-reducing and propionate-oxidizing bacteria and H2-consuming methanogens. Temperature is an important environmental regulator; yet its effect on syntrophic propionate oxidation has been poorly understood. In the present study, we investigated the syntrophic oxidation of propionate in a rice field soil at 15°C and 30°C. [U-13C]propionate (99 atom%) was applied to anoxic soil slurries, and the bacteria and archaea assimilating13C were traced by DNA-based stable isotope probing.Syntrophobacterspp.,Pelotomaculumspp., andSmithellaspp. were found significantly incorporating13C into their nucleic acids after [13C]propionate incubation at 30°C. The activity ofSmithellaspp. increased in the later stage, and concurrently that ofSyntrophomonasspp. increased. AceticlasticMethanosaetaceaeand hydrogenotrophicMethanomicrobialesandMethanocellalesacted as methanogenic partners at 30°C. Syntrophic oxidation of propionate also occurred actively at 15°C.Syntrophobacterspp. were significantly labeled with13C, whereasPelotomaculumspp. were less active at this temperature. In addition,Methanomicrobiales,Methanocellales, andMethanosarcinaceaedominated the methanogenic community, whileMethanosaetaceaedecreased. Collectively, temperature markedly influenced the activity and community structure of syntrophic guilds degrading propionate in the rice field soil. Interestingly,Geobacterspp. and some other anaerobic organisms likeRhodocyclaceae,Acidobacteria,Actinobacteria, andThermomicrobiaprobably also assimilated propionate-derived13C. The mechanisms for the involvement of these organisms remain unclear.

Detection of active, potentially acetate-oxidizing syntrophs in an anaerobic digester by flux measurement and formyltetrahydrofolate synthetase (FTHFS) expression profiling

Syntrophic oxidation of acetate, so-called reversed reductive acetogenesis, is one of the most important degradation steps in anaerobic digesters. However, little is known about the genetic diversity of the micro-organisms involved. Here we investigated the activity and composition of potentially acetate-oxidizing syntrophs using a combinatorial approach of flux measurement and transcriptional profiling of the formyltetrahydrofolate synthetase (FTHFS) gene, an ecological biomarker for reductive acetogenesis. During the operation of a thermophilic anaerobic digester, volatile fatty acids were mostly depleted, suggesting a high turnover rate for dissolved H2, and hydrogenotrophic methanogens were the dominant archaeal members. Batch cultivation of the digester microbiota with 13C-labelled acetate indicated that syntrophic oxidation accounted for 13.1–21.3 % of methane production from acetate. FTHFS genes were transcribed in the absence of carbon monoxide, methoxylated compounds and inorganic electron acceptors other than CO2, which is implicated in the activity of reversed reductive acetogenesis; however, expression itself does not distinguish whether biosynthesis or biodegradation is functioning. The mRNA- and DNA-based terminal RFLP and clone library analyses indicated that, out of nine FTHFS phylotypes detected, the FTHFS genes from the novel phylotypes I–IV in addition to the known syntroph Thermacetogenium phaeum (i.e. phylotype V) were specifically expressed. These transcripts arose from phylogenetically presumed homoacetogens. The results of this study demonstrate that hitherto unidentified phylotypes of homoacetogens are responsible for syntrophic acetate oxidation in an anaerobic digester.