Total microbial activity and microbial composition of a mangrove sediment are reduced by oil pollution at a site in the Arabian Gulf

In a study carried out to determine the effect of oil pollution on the microbiota of sediment associated with mangroves in the United Arab Emirates, sediment samples were collected from oil-polluted and nonpolluted mangrove sites. The levels of the total recoverable hydrocarbons and the polycyclic aromatic hydrocarbons assayed were noticeably higher in the polluted sediment. Microbial activity as measured by the hydrolysis of fluorescein diacetate and by the total populations of the culturable aerobic and anaerobic bacteria, streptomycete and non-streptomycete actinomycetes, and filamentous fungi and yeasts was significantly (P < 0.05) lower in the polluted than in the nonpolluted sediment. The estimated total aerobic and anaerobic hydrocarbon-utilizing bacteria were significantly (P < 0.05) higher in the polluted than in the nonpolluted sediments. Four days after the addition of the water-soluble fractions of the light Arabian crude oil to the nonpolluted sediment, at 10 different concentrations, there was a significant (P < 0.05) reduction (65%) in the microbial activity of the sediment compared with that of nonamended sediment. Concentrations of water-soluble fractions at 0.1% and above significantly and progressively reduced microbial activity, with total cessation of activity recorded at levels >50%. This study is the first to evaluate the effect of oil pollution on aerobic and anaerobic microbial flora of sediment of mangrove communities.     

Different methods for extracting bacteria from freshwater sediment and a simple method to measure bacterial production in sediment samples

The efficiency of different treatments was tested to extract bacterial cells from freshwater sediment samples. The influence of sonication, density gradient centrifugation, fixation by formalin and centrifugation speed on bacterial recovery was investigated. The method developed by Smith and Azam [Mar. Microb. Food Webs 6 (1992) 107] to measure microbial activity on bacterioplankton (3H-leucine incorporation), was also evaluated in sediment samples. After 1 min of sonication bacterial abundance was reduced by about 47% in diluted sediments with tetrasodium pyrophosphate. With the addition of Percoll after sonication, bacterial counts were not significantly different (P<0.05). Fixation by formalin increased bacterial counts using sonication. However, higher bacterial abundance was estimated in non-sonicated samples. Bacterial abundance in samples centrifuged at 7000xg with and without Percoll was not significantly different (P<0.05). Highest bacterial abundance was obtained after centrifugation at low speed (750xg). Bacterial abundance decreased with higher centrifugation speed (750, 1500 and 3000xg), the difference, however, was not significant. Bacterial production ranged from 0.10 microg C cm(-3) d(-1) in autoclaved sediment to 0. 27 microg C cm(-3) d(-1) in untreated sediment. The radioactivity measured in controls of both untreated and autoclaved sediment was high (70 and 91%, respectively), indicating a high level of leucine adsorption in sediment particles. In contrast, radioactivity in control samples previously centrifuged was markedly lower (6%). Despite the high values of radioactivity in the controls, bacterial production in untreated sediment was significantly higher than in centrifuged sediment (P<0.05).     

Sediment resuspension effects on alkaline phosphatase activity

Sediment cores, including the associated lake water, were collected from a shallow hypereutrophic lake located in central Florida. Alkaline phosphatase activity (APA) was measured as an indicator of potential organic P mineralization. In both the sediment and water columns, APA was mainly associated with particulate matter; < 10% of APA was within the soluble phase. This suggests that for enzymatic hydrolysis to occur the hydrolyzable organic compounds must be in close proximity to the particle-bound enzyme complex. Both total P (TP) and APA decreased with depth in the sediment, whereas soluble reactive P increased in the 20–40 cm fraction. Resuspension of surficial sediments resulted in an immediate increase in APA, total suspended solids, TP, total Kjeldahl N, and total organic C within the overlying water column. However, these concentrations decreased rapidly following cessation of turbulence and settling of the sediments, emphasizing the close association of these parameters with the sediment.     

Anaerobic ammonium-oxidizing bacteria in marine environments: widespread occurrence but low diversity

Laboratory and field studies have indicated that anaerobic ammonium oxidation (anammox) is an important process in the marine nitrogen cycle. In this study 11 additional anoxic marine sediment and water column samples were studied to substantiate this claim. In a combined approach using the molecular methods, polymerase chain reaction (PCR), qualitative and quantitative fluorescence in situ hybridization (FISH), as well as (15)N stable isotope activity measurements, it was shown that anammox bacteria were present and active in all samples investigated. The anammox activity measured in the sediment samples ranged from 0.08 fmol cell(-1) day(-1) N(2) in the Golfo Dulce (Pacific Ocean, Costa Rica) sediment to 0.98 fmol cell(-1) day(-1) N(2) in the Gullmarsfjorden (North Sea, Sweden) sediment. The percentage of anammox cell of the total population (stained with DAPI) as assessed by quantitative FISH was highest in the Barents Sea (9% +/- 4%) and in most of the samples well over 2%. Fluorescence in situ hybridization and phylogenetic analysis of the PCR products derived from the marine samples indicated the exclusive presence of members of the Candidatus'Scalindua' genus. This study showed the ubiquitous presence of anammox bacteria in anoxic marine ecosystems, supporting previous observations on the importance of anammox for N cycling in marine environments.     

Heterotrophic Activity Throughout a Vertical Profile of Seawater and Sediment in Halifax Harbor, Canada

The relative heterotrophic activity of marine microorganisms was determined at two sites by the heterotrophic uptake technique throughout the water column, the sediment-water interface, and the surface layer of sediment. In the water column, uptake was greatest at the surface and steadily decreased with depth. The percentage of the substrate that was respired also decreased with depth from 69 to 56%. The activity of the sediment-water interface was several orders of magnitude greater than that of the overlying water and twice that of the sediment immediately below. Hand-collected water samples carefully taken as close as 1 cm from the sediment-water interface had the same characteristically low activity as the bottom few meters of water. Microautoradiography with ³H-labeled glucose, glutamic acid, or thymidine revealed a general decrease in the percentage of active cells with depth from 35 to <1%. The number of active cells in the interface and sediment averaged <10% of the total population. The data indicate that the sediment-water interface is the most active region in this system due to an increased number of active cells rather than an increased percentage of active cells or increased per-cell activity.     

Influence of Deep Ocean Sewage Outfalls on the Microbial Activity of the Surrounding Sediment

The microbial activity near two deep ocean sewage outfalls off the coast of the island of Oahu, Hawaii, was characterized. Water samples and sediment samples to a depth of 4.5 cm were analyzed from an area of approximately 4.5 × 10⁴ m² surrounding the outfalls. Although the effluent water at both sites exhibited heterotrophic activity that was 2 orders of magnitude greater than water from a control site, ambient water samples taken within 1 m of the discharge ports exhibited activity only twice that of the control water. The heterotrophic activity of the outfall sediment was only elevated above that of the control site for surface samples collected within 10 m of the outfall. Likewise, the rates of microbial nucleic acid synthesis and carbon production in the sediment were only elevated immediately adjacent to the outfalls. Total microbial biomass, as determined by the ATP content of the sediment, varied spatially but was generally elevated at the outfall sites. The specific growth rates calculated for the sediment microbial populations, however, were not greater at the outfall sites. At one site the rocks surrounding the diffuser pipe were covered with copious amounts of slime that appeared to be composed entirely of microbial cells and filaments. This microbial mat was extremely active with respect to heterotrophic activity and biomass production. Overall, it appears that the impact of the sewage discharge on the ambient seawater microbiota is slight and that the effect on the sediment microbiota is confined to an area immediately adjacent to the diffuser ports. In the sand itself, the effect is limited to the upper 2 cm at most.     

水体沉积物中酸可挥发性硫化物(AVS)研究进展

水体沉积物中酸可挥发性硫化物 (AVS)是总硫含量中活性最高的部分 ,是沉积物中有毒重金属的重要结合形态 ,它的含量在很大程度上影响着沉积物重金属的生物有效性 ,从而作为沉积物中有毒重金属环境污染评价的一个重要指标 ;就十多年来水体沉积物中酸可挥发性硫化物 (AVS)的研究进行了综述。概述了 AVS的测定方法及其影响因素 ;探讨了水体沉积物中 AVS含量时空变化的规律 ;同时就目前“同时可提取重金属”(SEM)与 AVS摩尔浓度比值和水体沉积物重金属生物毒性关系的研究进行了概括和分析。     

Inhibition of methanogenesis by ethylene and other unsaturated hydrocarbons

Abstract Ethylene (ethene) was found to inhibit methane formation in slurries from sewage sludge and sediment samples taken from freshwater and marine sources. Methane formation from sediment contents was inhibited by 50% at 0.07% ethylene concentration in the gas phase (approx. 5 μmol · 1⁻¹ in the aqueous phase) and by 94% at ≥0.05% ethylene in the gas phase (≥36 μ mol · 1⁻¹ in the aqueous phase). Sulphate reduction was not impaired. Methane formation from added acetate, hydrogen or methanol was inhibited by ≥98%, from lactate by about 90%. The inhibition was reversible, and methanogenic activity recuperated completely after ethylene removal. Cyclopentadiene and cycloheptatriene led to strong inhibition; benzene, toluene, isoprene, and 1-hexine to moderate inhibition of methanogenesis; several unsaturated linear hydrocarbons were without effect. Pure cultures of Methanospirillum hungatei, Methanothrix soehngenii , and Methanosarcina barkeri were all inhibited by 50% at 0.05–0.1% ethylene concentration in the gas phase (3.6–7.2 μmol · 1⁻¹ in the aqueous phase). Pure cultures of Acetobacterium woodii, Halobacterium halobium and Sulfolobus acidocaldarius were not significantly inhibited by either ethylene or acetylene. Ethylene is recommended as a selective inhibitor of methanogenesis for physiological and enrichment experiments with sediment and sludge samples.     

Iron:phosphorus ratio in surface sediment as an indicator of phosphate release from aerobic sediments in shallow lakes

Analysis of Danish lakes showed that both mean winter and mean summer concentrations of lake water total phosphorus in the trophogenic zone correlated negatively with the total iron to total phosphorus ratio (Fe:P) in surface sediments. No correlation was found between the water total phosphorus concentration and either the sediment phosphorus concentration alone or with sediment calcium concentration. The increase in total phosphorus from winter to summer, which is partly a function of net internal P-loading, was lowest in lakes with high Fe:P ratios in the surface sediment.A study of aerobic sediments from fifteen lakes, selected as representative of Danish lakes with respect to the sediment Fe and phosphorus content, showed that the release of soluble reactive phosphorus was negatively correlated with the surface sediment Fe:P ratio. Analysis of phosphate adsorption properties of surface sediment from 12 lakes revealed that the capability of aerobic sediments to buffer phosphate concentration correlated with the Fe:P ratio while the maximum adsorption capacity correlated with total iron. Thus, the Fe:P ratio may provide a measure of free sorption sites for orthophosphate ions on iron hydroxyoxide surfaces.The results indicate that provided the Fe:P ratio is above 15 (by weight) it may be possible to control internal P-loading by keeping the surface sediment oxidized. Since the Fe:P ratio is easy to measure, it may be a useful tool in the management of shallow lakes.     

Short-term variations in the fluxes and composition of seston in near-bottom traps in the southern North Sea

In this study we attempted to measure the potential food availability for macrobenthic interface feeders using a new device (sediment recorder) which traps bed load particles at a height of 10 cm above the sea floor. The recorder is positioned flat on the seabed and alternately exposes a shallow collector mounted in a smooth surface mimicking the sediment surface. The samples were analyzed for their content of chlorophyll a (Chl) and dry weight (DM). We compared the sediment recorder data with the results obtained with a sediment trap suspended 3.2 m above the sea floor. Measurements were carried out in spring and fall at two stations in the southern North Sea with different sediment type, maximum current velocities and biomass (sand, station B vs silty sand, station F). The recorder deployments in November yielded higher Chl fluxes and a higher quality of particles (Chl:DM ratio) than the trap at both stations. Unlike the trap samples, in which Chl and DM were always positively and strongly correlated, maximum Chl and DM fluxes in the recorder samples were out of phase, particularly at station B, pointing to a highly variable quality during the tidal cycle. Unexpectedly, Chl fluxes and Chl:DM ratios in the recorder samples at station B were higher than at station F in both seasons, although the benthic biomass at the latter station was five times higher. The highest Chl fluxes and particle quality (Chl:DM) at station B were measured during slack tides when interface feeding would be least hampered by drag forces. Station B is nevertheless characterized by a low macrobenthic biomass and a scarcity of interface feeders, specifically of suspension-feeding bivalves. Possible reasons for this apparent discrepancy are discussed. Electronic Publication     

渤海沉积物产脂肪酶细菌的筛选及其多样性分析

【目的】从渤海沉积物中分离筛选产脂肪酶细菌,分析其物种多样性,增加人们对渤海生态系统中产脂肪酶菌多样性的认识,获取高效产脂肪酶菌株,为海洋产脂肪酶微生物的挖掘提供菌群资源。【方法】分别将8个渤海沉积物样品梯度稀释涂布至吐温-80筛选平板和三丁酸甘油酯筛选平板,选择性分离产脂肪酶细菌;分析基于16SrRNA基因序列的系统发育关系,揭示这些细菌的分类地位和遗传多样性;利用对硝基苯酚法测定胞外脂肪酶活性,筛选出高效产脂肪酶菌株。【结果】从8个渤海沉积物样品中分离获得51株产脂肪酶细菌,这些菌株隶属于Bacteroidetes、Proteobacteria和Firmicutes三个门的8个属,其中Pseudoalteromonas(35.2%)、Marinobacter(23.5%)和Sulfitobacter(17.6%)是优势菌群;脂肪酶酶活性实验表明所有测定菌株都能够分泌脂肪酶,菌株70623分泌的脂肪酶酶活最高,为42.4 U/m L。【结论】渤海沉积物中可培养产脂肪酶细菌类群较为丰富,Pseudoalteromonas、Marinobacter和Sulfitobacter菌株是优势菌群,测定菌株所产胞外脂肪酶能力不同,获得了一株高效产脂肪酶菌株Marinobacter sp.70623。     

Prokaryotic metabolic activity and community structure in Antarctic continental shelf sediments

The prokaryote community activity and structural characteristics within marine sediment sampled across a continental shelf area located off eastern Antarctica (66 degrees S, 143 degrees E; depth range, 709 to 964 m) were studied. Correlations were found between microbial biomass and aminopeptidase and chitinase rates, which were used as proxies for microbial activity. Biomass and activity were maximal within the 0- to 3-cm depth range and declined rapidly with sediment depths below 5 cm. Most-probable-number counting using a dilute carbohydrate-containing medium recovered 1.7 to 3.8% of the sediment total bacterial count, with mostly facultatively anaerobic psychrophiles cultured. The median optimal growth temperature for the sediment isolates was 15 degrees C. Many of the isolates identified belonged to genera characteristic of deep-sea habitats, although most appear to be novel species. Phospholipid fatty acid (PLFA) and isoprenoid glycerol dialkyl glycerol tetraether analyses indicated that the samples contained lipid components typical of marine sediments, with profiles varying little between samples at the same depth; however, significant differences in PLFA profiles were found between depths of 0 to 1 cm and 13 to 15 cm, reflecting the presence of a different microbial community. Denaturing gradient gel electrophoresis (DGGE) analysis of amplified bacterial 16S rRNA genes revealed that between samples and across sediment core depths of 1 to 4 cm, the community structure appeared homogenous; however, principal-component analysis of DGGE patterns revealed that at greater sediment depths, successional shifts in community structure were evident. Sequencing of DGGE bands and rRNA probe hybridization analysis revealed that the major community members belonged to delta proteobacteria, putative sulfide oxidizers of the gamma proteobacteria, Flavobacteria, Planctomycetales, and Archaea. rRNA hybridization analyses also indicated that these groups were present at similar levels in the top layer across the shelf region.     

Optimization of direct cell counting in sediment

This study reports a method for optimizing direct counts of bacteria in sediment, designed to reduce the masking by sediment particles. The protocol was designed to determine appropriate dilution factors by incorporating counting statistics and was used to measure depth-associated changes in microbial abundance in metal-impacted freshwater sediments. We demonstrated a direct method to determine appropriate sample dilution for accurate counting by adding a known amount of cells to the sediment. For accurate counting in our sediment samples, we determined that the average number of bacteria per microscope ocular field must be between 8.5 and 10. This is well below the 30 bacteria/field previously suggested for accurate counting. These results indicate that an optimal dilution rate must be determined before accurate direct counts in sediment can be achieved.     

Preconcentration of pharmaceuticals residues in sediment samples using microwave assisted micellar extraction coupled with solid phase extraction and their determination by HPLC-UV

An analytical method combining microwave assisted micellar extraction (MAME) and solid phase extraction (SPE) has been developed to extract and preconcentrate a selected group of eight pharmaceutical compounds in sediment samples prior to their determination using liquid chromatography with an UV-DAD detector. A non-ionic surfactant, Polyoxyethylene 10 lauryl ether (POLE) was used for the MAME extraction and the different parameters for the optimization process were studied. Then, SPE was used to clean-up and preconcentrate the target analytes in the extract, prior to their determination using HPLC-UV. The method was applied to the determination of the selected pharmaceuticals compounds in several kinds of sediment samples with different characteristics. Relative recoveries for spiked sediment samples were over 70% and relative standard deviations (RSDs) were under 11% for all recoveries tested. Detection limits between 4 and 167ngg(-1) were obtained. The method was validated using Soxhlet extraction procedure.     

Assessing microbial diversity using recent lake sediments and estimations of spatio‐temporal diversity

Aim Recent papers have used large palaeolimnological datasets to reveal the biodiversity patterns of aquatic microorganisms. However, scant attention has been paid to the influence of time on these patterns. Where lake surficial sediment samples are used as integrals of diversity, the time interval of each sample varies according to differences in sediment accumulation rates. This paper aims to test the reliability of using lake surface sediments to measure and to compare microbial diversity when the potential influences of the species–time relationships are taken into account. Location Alpine lakes in Europe. Methods We analysed microorganism (siliceous microalgae) assemblages in three European Alpine lakes using short sediment cores (²¹⁰Pb‐dated) and annual sediment trap samples from 12 UK lakes. The same number of individuals was pooled for each sample 500 times to avoid sampling effort effects and to standardize species diversity estimation. The influence of time on the diversity score was assessed by simulating an increase of time span for surface sediment samples by cumulatively adding in successive sediment core samples (from the most recent to the oldest). We used species richness (S) and the exponential of the bias‐corrected Shannon entropy index (exp(H_b‐c)) to estimate diversity. Results Increasing the time interval represented by a surficial sediment sample did not affect the diversity results. The estimation of diversity was similar for cumulative and non‐cumulative samples. Diversity estimation was only altered in lakes experiencing high community turnover due to strong environmental forcing during the time period spanned by the cumulative sample. Main conclusions The use of surface lake sediments is suitable for estimating the average site diversity of free‐living microorganisms. Diversity is integrated in a single sample and species assemblage composition is derived from microbial communities living in distinct lake microhabitats. Species remains, accumulated in a single sample over several years of environmental variability, represent a diversity integral that captures a spatio‐temporal component equivalent to the γ‐diversity measure.     

A Sensitive Method Using 4-Methylumbelliferyl-beta-Cellobiose as a Substrate To Measure (1,4)-beta-Glucanase Activity in Sediments

A sensitive method to measure (1,4)-beta-glucanase activity in organic matter-rich sediments, using 4-methyl-umbelliferyl-beta-cellobiose as a substrate, is described. beta-Glucosidases, which were also able to hydrolyze this substrate, were inhibited with d-glucono-delta-lactone. The produced 4-methylumbelliferone was recovered quantitatively out of the sediment by an extraction with 80% ethanol. An inhibition experiment with known substrates or inhibitors suggested that at least 59% of the measured activity could be explained by enzymes of the exo-(1,4)-beta-glucanase type and that the contribution of endo-(1,4)-beta-glucanases was minor. Results of the inhibition experiment also suggested that the measured activity was of bacterial origin in the sediment used. First results of field measurements are given for the sediment from the reed bed of Lake Gooimeer.     

Temporal Variations in Heterotrophic Mesophilic Bacteria from a Marine Shallow Hydrothermal Vent off the Island of Vulcano (Eolian Islands, Italy)

Abstract The fluctuations of the total microbial abundance, the culturable heterotrophic bacterial population, and the composition of heterotrophic bacteria were investigated in relation to environmental parameters in a shallow, marine hydrothermal vent off the Island of Vulcano (Eolian Islands, Italy). Standing stock dynamics were studied by measuring the total population of picoplankton by direct count and the population of viable heterotrophic bacteria in water and sediment samples collected monthly. The environmental factors most strongly linked to the total microbial abundance and heterotrophic bacterial populations were pH and H₂S content in water and C/N ratio in sediment samples. The pattern of variation of microbial populations associated with water was different from those associated with sediment. Assessment of the qualitative composition of aerobic heterotrophic bacterial communities was based on 30 morphological and biochemical characteristics for each strain. Numerical analysis was used for an initial survey of the similarity among the isolates. The data were successively used to determine the structure and the metabolic potential of water and sediment bacteria. Metabolic properties varied between water- and sediment-isolated bacteria. Bacteria from water were structurally more diverse, and active in the use of carbohydrates, than those from sediment. Moreover, most of the sediment bacteria were able to grow at a high temperature (60 and 70°C). The fluctuations of bacterial characteristics in relation to environmental parameters present an evident temporal variation in water, but not in the sediment habitat. Received: 13 January 1997; Accepted: 7 August 1997     

黄河三角洲滨岸潮滩湿地碱蓬对水沙条件及氮输入的适应性

调水调沙工程的实施为黄河三角洲带来大量淡水恢复退化湿地的同时,也改变了湿地的水沙状况并带来大量的外源物质。本研究采用3因素4水平的正交试验,以黄河口滨岸潮滩湿地典型的先锋物种碱蓬为研究对象,探讨了其对淹水深度、泥沙埋深及外源氮输入的适应机制。结果表明: 泥沙埋深对碱蓬叶片蛋白含量及超氧化物歧化酶(SOD)活性的影响较大;氮输入对过氧化物酶(POD)活性的影响较大,而3种处理对过氧化氢酶(CAT)活性的影响无显著差异。淹水深度对叶、茎及总干重具有显著影响,且随淹水深度的增加呈现减小趋势,最大值(25.70、40.86、69.73 g)均出现在2 cm淹水处理;而氮输入及泥沙埋深对碱蓬干重的影响不显著。极差分析表明,淹水深度对碱蓬叶、茎、根及总干重的影响最为显著,其次为氮输入和泥沙埋深,2 cm淹水深度+12 cm埋深深度+9 g·m^-2氮输入量最有利于碱蓬的生长。可见,淹水深度的变化对碱蓬的生长起到决定性的作用,在调水调沙等工程的实施过程中应注重淹水深度的把控。     

Sulfhydrolase Activity in Sediments of Wintergreen Lake, Kalamazoo County, Michigan

The hydrolysis of p-nitrophenyl sulfate, p-nitrocatechol sulfate, and [³⁵S]sodium dodecyl sulfate was examined in anoxic sediments of Wintergreen Lake, Michigan. Significant levels of sulfhydrolase activity were observed in littoral, transition, and profundal sediment samples. Rates of sulfate formation suggest that the sulfhydrolase system would represent a major source of sulfate within these sediments. Sulfate formed by ester sulfate hydrolysis can support dissimilatory sulfate reduction as shown by the incorporation of ³⁵S from labeled sodium dodecyl sulfate into H₂³⁵S. Sulfhydrolase activity varied with sediment depth, was greatest in the littoral zone, and was sensitive to the presence of oxygen. Estimations of ester sulfate concentrations in sediments revealed large quantities of ester sulfate (~30% of total sulfur). Both total sulfur and ester sulfate concentrations varied with the sediment type and were two to three orders of magnitude greater than the inorganic sulfur concentration.     

Hydrogenase Activity in Deeply Buried Sediments of the Arctic and North Atlantic Oceans

The goal of this study was to measure hydrogenase activity in oceanic sediments of the Lomonosov Ridge (Arctic Ocean) and of the Porcupine Seabight (North Atlantic), and to understand how its distribution varied depending on geochemical and lithological characteristics of the sediment. Hydrogenase activity was found at all sites, and absolute values and downhole distributions varied widely within and between sites. At the Lomonosov Ridge, hydrogenase was below detection in the top 190 meters of sediment, but high levels were measured in the organic carbon-rich layers below this depth. Activity at Challenger Mound, Porcupine Seabight (Site 1317) was one to two orders of magnitude higher than the upslope or downslope sites (1318 and 1316, respectively), and was higher in the mound than below the mound base. Consistent with the interpretation of hydrogenase as an indicator of microbial activity, cells were present in all hydrogenase-positive samples or in nearby horizons. Cell-specific activity values were as much as 1000 fold lower than those of cultured Clostridium pasteurianum, with the values from the highest activity sediments approaching those of C. pasteurianum more closely. This suggests that little adaptation in hydrogenase activity might be necessary to support H₂ metabolism in deeply buried sediments that contain relatively high levels of substrates.