Growth dynamics of major microbial populations during biodegradation of o-phthalate in anaerobic sediment slurries under a CO2/H2 atmosphere

o-Phthalate transformers increased about five orders of magnitude (to 1.6x10(11)cells g-1 sediment) just before the onset of fast biotransformation of o-phthalate (21.6 mg l-1) and then decreased sharply when the concentration of o-phthalate became low during biodegradation of o-phthalate in anaerobic sediment slurries under CO2/H2 (4:1, v/v). In contrast, the benzoate transformers increased about four orders of magnitude (to 1.6x10(11)cells g-1 sediment) in 48 days and then increased one more order (to 1.6x10(12)cells g-1 sediment) in 60 days and then remained at that high level in those sediment slurries. When making a comparison between the growth dynamics of o-phthalate transformers, acetogens, sulfate reducers, and methanogens and the time course of o-phthalate transformation, it appears that acetogens did not initiate biotransformation of o-phthalate, and that sulfate reducers and methanogens were not directly involved in o-phthalte degradation. o-Phthalate was not transformed in sediment slurries amended with BESA plus molybdate under CO2/H2.     

Microbially mediated redox processes in natural analogues for radioactive waste

Natural analogues allow scientists to investigate biogeochemical processes relevant to radioactive waste disposal that occur on time scales longer than those that may be studied by time-limited laboratory experiments. The Palmottu U-Th deposit in Finland and the Bangombé natural nuclear reactor in Gabon involve the study of natural uranium, and are both considered natural analogues for subsurface radioactive waste disposal. The microbial population naturally present in groundwater may affect the redox conditions, and hence, the radionuclide solubility and migration. Therefore, groundwater samples from the two sites were investigated for microbial populations. The total numbers of cells ranged from 10(4) to 10(6) cells ml(-1). Iron-reducing bacteria (IRB) were the largest culturable microbial population in the Palmottu groundwater and were present at up to 1.3 x 10(5) cells ml(-1). Sulfate-reducing bacteria (SRB) and acetogens could also be cultured from the Palmottu groundwater. The numbers of IRB and SRB were largest in groundwater with the lowest uranium concentrations. Removal of dissolved U(VI) from solution was concomitant with the growth of IRB enrichment cultures and the reduction of iron. The redox buffer in the Palmottu groundwater consists of iron and uranium species, both of which are affected by IRB. IRB and aerobic heterotrophs were cultured from the Bangombé groundwater, where redox potentials are buffered by iron and organic carbon species. Microbial populations similar to those found at Palmottu and Bangombé are found throughout the Fennoscandian Shield, a potential host rock for subsurface radioactive waste disposal. These results confirm that microorganisms can be expected to play a role in stabilizing radioactive waste disposed of in the subsurface by lowering redox potential and immobilizing radionuclides.     

Column experiments to assess the effects of electron donors on the efficiency of in situ precipitation of Zn,Cd, Co and Ni in contaminated groundwater applying the biological sulfate removal technology

BACKGROUND, AIMS AND SCOPE: In a previous study, we explored the use of acetate, lactate, molasses, Hydrogen Release Compound (HRC, which is based on a biodegradable poly-lactate ester), methanol and ethanol as carbon source and electron donor to promote bacterial sulfate reduction in batch experiments, this with regards to applying an in situ metal precipitation (ISMP) process as a remediation tool to treat heavy metal contaminated groundwater at the site of a nonferrous metal work company. Based on the results of these batch tests, column experiments were conducted with lactate, molasses and HRCI as the next step in our preliminary study for a go-no go decision for dimensioning an on site application of the ISMP process that applies the activity of the endogenous population of sulfate-reducing bacteria (SRB). Special attention was given to the sustainability of the metal precipitation process under circumstances of changing chemical oxygen demand (COD) to [SO4(2-)] ratios or disrupted substrate supply. METHODS: To optimize the ISMP process, an insight is needed in the composition and activity of the indigenous SRB community, as well as information on the way its composition and activity are affected by process conditions such as the added type of C-source/ electron donor, or the presence of other prokaryotes (e.g. fermenting bacteria, methane producing Archaea, acetogens). Therefore, the biological sulfate reduction process in the column experiments was evaluated by combining classical analytical methods [measuring heavy metal concentration, SO4(2-)-concentration, pH, dissolved organic carbon (DOC)] with molecular methods [denaturing gradient gel electrophoresis (DGGE) fingerprinting and phylogenetic sequence analysis] based on either the 16S rRNA-gene or the dsr (dissimilatory sulfite reductase) gene, the latter being a specific biomarker for SRB. RESULTS AND DISCUSSION: All carbon sources tested promoted SRB activity, which resulted within 8 weeks in a drastic reduction of the sulfate and heavy metal contents in the column effluents. However, unexpected temporal decreases in the efficiency of the ISMP process, accompanied by the release of precipitated metals, were observed for most conditions tested. The most dramatic observation of the failing ISMP process was observed within 12 weeks for the molasses amended column. Subsequent lowering the COD/ SO4(2-) ratio from 1.9 to 0.4 did not alter the outcome of sulfate reduction and metal precipitation efficiency in this set-up. Remarkably, after 6 months of inactivity, bacterial sulfate reduction was recovered in the molasses set up when the original COD/ SO4(2-) ratio of 1.9 was applied again. Intentional disruption of the lactate and HRC supplies resulted in an immediate stagnation of the ISMP processes and in a rapid release of precipitated metals into the column effluents. However, the ISMP process could be restored after substrate amendment. 16S rDNA-based DGGE analysis revealed that the SRB population, in accordance with the results of the previously performed batch experiments, consisted exclusively of members of the genus Desulfosporosinus. The community of Archaea was characterized by sequencing amplicons of archaeal and methanogen-specific PCR reactions. This approach only revealed the presence of non-thermophilic Crenarchaeota, a novel group of organisms which is only distantly related to methane producing Euryarchaeota. DGGE on the dsrB genes was successfully used to link the results of the ISMP process to the community composition of the sulfate reducing bacteria. CONCLUSIONS: In the case of an intentional disruption of substrate supply, the ISMP process failed most likely because the growth and activity of the indigenous SRB community stopped due to a lack of a carbon and electron donor. On the other hand, the cause of the sudden temporal shortcomings of the ISMP process in the presence of different substrates was not immediately clear. It was first thought to be the result of competition between methanogenic prokaryotes (MP) and sulfate reducers, since the formation of small amounts of CH4 (0.01-0.03 ppm ml(-1) was detected. However, the results of molecular analyzes indicate that methanogens do not constitute a major fraction of the microbial communities that were enriched in the column experiments. Therefore, we postulate that the SRB population becomes inhibited by the formed metal sulfides. RECOMMENDATION AND PERSPECTIVE: Our results indicate that the ISMP process is highly dependent on SRB-stimulation by substrate amendments and suggest that this remedial approach might not be viable for long-term application unless substrate amendments are continued and environmental conditions are strictly controlled. This will include the removal of affected aquifer material from the metal precipitation zone at the end of the remediation process, or removal of metal precipitates when the microbial activity decreases. Additional tests are necessary to investigate what will happen when clear groundwater passes through the reactive zone while no more C-sources are amended and all indigenous carbon is consumed. Also, the effects of dramatic increases in sulfate- or HM-concentrations on the SRB-community and the concomitant ISMP process need to be studied in more detail.     

干式厌氧消化过程挥发酸对硫酸盐还原菌的影响

在牛粪干式厌氧消化过程中,通过添加不同挥发酸(乙酸、丙酸、丁酸),考察消化稳定阶段,挥发性脂肪酸的分布特征,挥发性脂肪酸酸组成变化对硫酸盐还原菌(SRB)的影响,微生物种群组成和种群间关系。实验结果表明,挥发性脂肪酸对SRB还原速率的贡献依次为:丙酸丁酸乙酸。相比乙酸和丁酸,添加一定量的丙酸,更有利于激活SRB的活性,从而加强SRB与产甲烷菌(MB)的种间协同,保证厌氧系统的稳定运行。     

Uptake and accumulation behaviour of angiosperms irrigated with solutions of different arsenic species

Uptake and metabolisation of arsenic as a function of both the plant type and the chemical form of arsenic were examined. For this purpose two different plant species (Silene vulgaris and Plantago major) were selected that differed in their vitality and accumulation behaviour on arsenic-loaded substrates. The plants were cultivated on soil and irrigated with aqueous solutions of an inorganic arsenic compound (arsenious acid) and an organic compound (dimethylarsinate). The arsenic species accumulated in the parts of the plants above ground were extracted by PLE and determined using IC-ICP-MS. The concentrations and metabolisation products of arsenic found in the extracts indicate different mechanisms of arsenic uptake and transformation in both angiosperms. The arsenic species pattern showed that S. vulgaris was more arsenic--tolerable than P. major which is attributed to a low arsenate to arsenite concentration ratio in the plant compartments. S. vulgaris was also able to demethylate and reduce dimethylarsinate to form arsenite in a high extent. P. major accumulated only eight times lower concentration of arsenic, and the arsenate to arsenite concentration ratio shifted to higher values. Metabolisation products of dimethylarsinate did not occur under the present experimental conditions. The vitality of the angiosperms seems to be very dependent on the ability of the plant to reduce arsenate to arsenite.     

Performance of various sub-grid scale models in large-eddy simulations of turbulent flow over complex terrain

Turbulent flow over a two-dimensional steep hill was analyzed by large-eddy simulations (LES). Here, six LES computations were carried out using four different sub-grid scale (SGS) models and two different ground surface conditions. The accuracy of these computations was assessed by comparing the results with those from an experiment by Ishihara et al. (An experimental study of turbulent boundary layer over a steep hill, Proceedings of the 15th National Symposium on Wind Engineering, 1998, pp. 61–66; J. Wind Eng. 89 (2001) 573). The results of the dynamic SGS models were in very poor agreement with those of the experiment. The poor prediction accuracy was mainly caused by the inaccurate estimation of the model coefficient near the ground surface. In order to improve the prediction accuracy of the dynamic SGS models, a hybrid SGS model, i.e., a combination of the standard Smagorinsky model and the dynamic Smagorinsky model, was introduced. The hybrid model provided very accurate predictions and produced the best results of the four SGS models compared here.     

Simulated and experimental evaluation of factors affecting the rate and extent of reductive dehalogenation of chloroethenes with glucose

Carbohydrates such as molasses are being added to aquifers to serve as electron donors for reductive dehalogenation of chloroethenes. Glucose, as a model carbohydrate, was studied to better understand the processes involved and to evaluate the effectiveness for dehalogenation of different approaches for carbohydrate addition. A simulation model was developed and calibrated with experimental data for the reductive dehalogenation of tetrachloroethene to ethene via cis-1,2-dichloroethene. The model included fermentors that convert the primary donor (glucose) into butyrate, acetate and hydrogen, methanogens, and two separate dehalogenator groups. The dehalogenation groups use the hydrogen intermediate as an electron donor and the different haloethenes as electron acceptors through competitive inhibition. Model simulations suggest first that the initial relative population size of dehalogenators and H(2)-utilizing methanogens greatly affects the degree of dehalogenation achieved. Second, the growth and decay of biomass from soluble carbohydrate plays a significant role in reductive dehalogenation. Finally, the carbohydrate delivery strategies used (periodic versus batch addition and the time interval between periodic addition) greatly affect the degree of dehalogenation that can be obtained with a given amount of added carbohydrate.     

Toxic effect of surfactants and probable products of their biodegradation on methanogenesis in an anaerobic microbial community.

Surfactants used in household and various industries, are rather toxic; therefore, the accumulation of these compounds in the environment through wastewaters has challenged the problem of their biodegradation. In this research, an attempt was made to assess the toxic effect of various surfactants and the likely products of their biodegradation on the acetoclastic methanogens of an anaerobic microbial community. Among the substances investigated, cationic surfactants were found to be most toxic to methanogens: 154 mg/l alkamon DS and 345 mg/l catamin AB induced a 50% inhibition of methanogenesis. Toxicity studies of some aromatic and cyclic compounds, as the probable products of biodegradation of alkylbenzene sulfonate surfactants, showed that methanogenesis in the microbial community under study are rather tolerant to high concentrations of these compounds.     

Precipitation of Zn(II), Cu(II) and Pb(II) at bench-scale using biogenic hydrogen sulfide from the utilization of volatile fatty acids

Biological production of hydrogen sulfide (H(2)S) using sulfate-reducing bacteria (SRB) has important potential within environmental biotechnology. The aim of this work was to study the possibility of using SRB for the treatment of an acid mine drainage (AMD) at bench-scale. This process involved three stages: the optimization of H(2)S production through the utilization of total volatile fatty acids (TVFAs) by SRB, the establishment of a biofilm reactor for sulfide production, and the precipitation of metals by using the biologically produced H(2)S. The substrates used for TVFAs production consisted of papaya, apple and banana. The H(2)S produced from the degradation of TVFAs was utilized for the precipitation of a metal-contaminated effluent collected from Bolivar mine (Oruro, Bolivia). The maximum concentration of H(2)S obtained was approximately 16mM. Removal efficiencies of ca. 100% for copper, above 94% for zinc, and above 92% for lead were achieved.     

Selenium oxyanion inhibition of hydrogenotrophic and acetoclastic methanogenesis

Inhibitory effects of selenite and selenate towards hydrogenotrophic and acetoclastic methanogenesis were evaluated in anaerobic toxicity assays. The 50% inhibitory concentration (IC(50)) of both selenium oxyanions was below 6.1x10(-5)M in hydrogenotrophic assays, whereas acetoclastic methanogens were less inhibited: IC(50)=8.3x10(-5)M and 5.5x10(-4)M for selenite and selenate, respectively. Selenite completely inhibits methanogenesis from both substrates tested at concentrations 10(-3)M selenite, while only marginal methanogenic activities occur at equimolar concentrations of selenate. Selenite becomes irreversibly inhibitory upon a single exposure, whereas selenate inhibits methanogens upon repeated exposure. Consequently, methane recovery can be seriously hampered or even impossible during anaerobic treatment of highly selenium contaminated waste streams.     

Anaerobic treatment of high-saline wastewater using halophilic methanogens in laboratory-scale anaerobic filters.

The presence of a high concentration of sodium in wastewater is considered inhibitory for anaerobic biological treatment. This research was designed to investigate the potential use of halophilic methanogens and a mixed culture of halophilic methanogens and digester sludge, in anaerobic filters, for treatment of organic pollutants in high-saline wastewater at 35 degrees C. Data related to startup of the filters are presented. Both halophilic and mixed-culture anaerobic filters were able to operate at a sodium chloride concentration of 35 g/L, at organic loading rates (OLRs) of 6.2 and 5 kg chemical oxygen demand (COD)/m(3) x d, respectively. The COD removal efficiency was as high as 80%, and the systems were able to maintain a low volatile fatty acids concentration of 500 mg/L. No significant difference in COD removal was observed between the halophilic filter and the mixed-culture filter. Increasing the salt concentration to 37 g/L at an OLR of 3 kg/m(3) x d caused system failure.     

Contributions of fermentative acidogenic bacteria and sulfate-reducing bacteria to lactate degradation and sulfate reduction

The roles of fermentative acidogenic bacteria and sulfate-reducing bacteria (SRB) in lactate degradation and sulfate reduction in a sulfidogenic bioreactor were investigated by traditional chemical monitoring and culture-independent methods. A continuously stirred tank reactor fed with synthetic wastewater containing lactate and SO(2-)(4) at 35 degrees C, 10h of hydraulic retention time was used. The results showed that sulfate removal efficiency reached 99%, and sulfide and acetate were the main end products after 20 d of operation. 16S rRNA gene based clone libraries and single-strand conformation polymorphism profiles demonstrated that the proportion of SRB increased from 16% to 95%, and that Desulfobulbus spp., Desulfovibrio spp., Pseudomonas spp. and Clostridium spp. formed a stable, dominant community structure. The decreasing COD/SO(2-)(4) ratio had little effect on the community pattern except that Pseudomonas spp. and Desulfobulbus spp. increased slightly. The addition of molybdate to the influent significantly changed the microbial community, sulfate removal efficiency and the pattern of end products. Clostridium spp., Bacteroides spp. and Ruminococcus spp. became the dominant community members. The main end products switched from acetate to ethanol and then to propionate with the oxidation-reduction potentials increasing from -420 to -290 mV. A lactate degradation pathway was deduced: lactate served as the electronic donor for Desulfovibrio spp., or was fermented by Clostridium spp. and Bacteroides spp. to produce propionate or ethanol, which were subsequently utilized by Desulfobulbus spp. and Desulfovibrio spp. The acidotrophic SRB oxidized part of the acetate finally.     

不同基质在人工湿地脱氮中的应用及其研究进展

在比较了不同基质脱氮效率的基础上,认为基质作为人工湿地的重要组成部分,在为植物和微生物提供生长介质的同时,也能通过沉淀、过滤和吸附等作用直接去除污染物质,其中基质的类型、级配等因素会影响基质作用的发挥;不同基质对脱氮性能存在较大差异,沸石和蛭石是目前研究中脱氮效率较高的两种基质。在归纳了脱氮机制和影响因素的基础上,认为不同基质由于脱氮机制不同,脱氮性能和脱氮效率也存在较大差异;人工湿地基质所有理化性状都可能影响到它对污水的脱氮效率。最后,对今后的相关研究方向进行了展望。     

Kinetics of phenol and chlorophenol utilization by Acinetobacter species

Although microbial transformations via cometabolism have been widely observed, the few available kinetic models of cometabolism have not adequately addressed the case of inhibition from both the growth and nongrowth substrates. The present study investigated the degradation kinetics of self-inhibitory growth (phenol) and nongrowth (4-chlorophenol, 4-CP) substrates, present individually and in combination. Specifically, batch experiments were performed using an Acinetobacter isolate growing on phenol alone and with 4-CP present. In addition, batch experiments were also performed to evaluate the transformation of 4-CP by resting, phenol-induced Acinetobacter cultures. The Haldane kinetic model adequately predicted the biodegradation of phenol alone, although a slight discrepancy was noted in cases of higher initial phenol concentrations. Similarly, a Haldane model for substrate utilization was also able to describe the trends in 4-CP transformation by the resting cell cultures. The 4-CP transformation by the Acinetobacter species growing on phenol was modeled using a competitive kinetic model of cometabolism, which included growth and nongrowth substrate inhibition and cross-inhibition terms. Excellent agreement was obtained between the model predictions using experimentally estimated parameter values and the experimental data for the synchronous disappearance of phenol and 4-CP.     

湿地稻-鸭复合生态种养对甲烷菌种群数量影响的研究

采用厌氧培养箱技术．用最大或然数计数(MPN)法测定稻一鸭复合种养对水稻不同生育期土壤中的产甲烷菌种群数量的影响。结果表明,稻田土壤中产甲烷菌数量的变化规律与稻田甲烷排放的季节变化规律基本一致,早稻田土壤中菌数高峰期出现在孕穗期．晚稻田土壤中菌数高峰期出现在分蘖盛期。实行稻-鸭复合种养,显著降低了稻田土壤中甲烷细菌种群的数量。特别是减少了甲烷排放高峰期的甲烷细菌憨量。早稻田和晚稻田产甲烷菌种群数量分别比对照田降低20%-96．9％和94．3％～95．8％,且两系统土壤中甲烷细菌数量的差异达1％的显著水平。     

微氧折流反应器中产甲烷菌的分布与群落结构分析

为了研究微氧条件下折流反应器处理生活污水的微生物学机理,针对产甲烷菌的MCR基因采用实时荧光定量PCR和构建克隆文库的方法,对反应器中的关键功能微生物产甲烷菌的分布特点与群落结构变化进行了分析。结果表明,微氧条件下折流反应器的环境条件可以保证产甲烷菌的增长,不同格室的产甲烷菌丰度均要高于接种污泥(1 720 copies/ng),其中经过微氧曝气处理的1#(2 864 copies/ng)和2#(2 282 copies/ng)格室产甲烷菌丰度低于厌氧状态的3#(3 508copies/ng)格室;微氧处理后折流反应器产甲烷菌的群落结构发生了明显的变化,接种污泥的产甲烷菌多属于Methanomicrobiales目的 Methanoregulaceae属(6/9),而保持微氧状态的1#格室中产甲烷菌多隶属于Methanomicrobiales目(6/10)的其他类群,2#格室的产甲烷菌则多隶属于Methanobacteriales目(5/10),保持厌氧状态的3#格室的产甲烷菌主要属于Methanomicrobiales目(3/10)和Methanosarcinales目(3/10);产甲烷菌MCR基因的多样性指数表明微氧曝气的引入降低了活性污泥中产甲烷菌的生物多样性。     

Characterization and reactivity assessment of organic substrates for sulphate-reducing bacteria in acid mine drainage treatment

Acid mine drainage (AMD), which contains high concentrations of sulphate and dissolved metals, is a serious environmental problem. It can be treated in situ by sulphate reducing bacteria (SRB), but effectiveness of the treatment process depends on the organic substrate chosen to supply the bacteria's carbon source. Six natural organic materials were characterized in order to investigate how well these promote sulphate reduction and metal precipitation by SRB. Maple wood chips, sphagnum peat moss, leaf compost, conifer compost, poultry manure and conifer sawdust were investigated in terms of their carbon (TOC, TIC, DOC) and nitrogen (TKN) content, as well as their easily available substances content (EAS). Single substrates, ethanol, a mixture of leaf compost (30% w/w), poultry manure (18% w/w), and maple wood chips (2% w/w), and the same mixture spiked with formaldehyde were then tested in a 70-day batch experiment to evaluate their performance in sulphate reduction and metal removal from synthetic AMD. Metal removal efficiency in batch reactors was as high as 100% for Fe, 99% for Mn, 99% for Cd, 99% for Ni, and 94% for Zn depending on reactive mixtures. Early metal removal (0-12d) was attributed to the precipitation of (oxy)hydroxides and carbonate minerals. The lowest metal and sulphate removal efficiency was found in the reactor containing poultry manure as the single carbon source despite its high DOC and EAS content. The mixture of organic materials was most effective in promoting sulphate reduction, followed by ethanol and maple wood chips, and single natural organic substrates generally showed low reactivity. Formaldehyde (0.015% (w/v)) provided only temporary bacterial inhibition. Although characterization of substrates on an individual basis provided insight on their chemical make-up, it did not give a clear indication of their ability to promote sulphate reduction and metal removal.     

Degradation of nitrogen containing organic compounds by combined photocatalysis and ozonation

The combination of TiO2-assisted photocatalysis and ozonation in the degradation of nitrogen-containing substrates such as alkylamines, alkanolamines, heterocyclic and aromatic N-compounds has been investigated. A laboratory set-up was designed and the influence of the structure of the N-compound, the TiO2 and ozone concentration on the formation of breakdown products were examined. The experimental results showed that a considerable increase in the degradation efficiency of the N-compounds is obtained by a combination of photocatalysis and ozonation as compared to either ozonation or photocatalysis only. The mineralization of the model substances was monitored by measurements of the TOC and ion-chromatographic determinations of the formed NO2- and NO3-. The temporal changes of concentrations of breakdown products, such as NH4+, short chain alkyl- and alkanolamines were determined by single column ion chromatography (SCIC) and as well as by electrospray mass spectrometry (EI-MS).     

Control of metal toxicity, effluent COD and regeneration of gel beads by immobilized sulfate-reducing bacteria

Over the last few decades, the use of sulfate-reducing bacteria (SRB) in the treatment of heavy-metal containing wastewaters including acid mine drainage has become a topic of scientific and commercial interest. However, technical difficulties such as the sensitivity of SRB to toxic metals and high effluent COD limit the widespread use of SRB in high heavy-metal containing wastewater. The aim of this study was to clarify the reasons why the immobilized SRB sludge with inner cohesive carbon source (ISIS) process can endure high metal toxicity and decrease effluent COD. The ISIS process can physically set apart SRB and free the system of external influences such as the surrounding toxic metallic ions, as well as form inner carbon sources to avoid high effluent COD. Metal toxicity and bead durability are the two major factors which influence the regeneration and reuse of gel beads. Reuse of suspended SRB sludge and beads crosslinked with boric acid were unsuccessful due to metal toxicity and agglomeration of beads, respectively. However, beads crosslinked with ammonium sulfate prevented agglomeration of beads allowing successful bead regeneration and reuse. The result of four cyclic trials showed that over 99% of zinc was removed in each trial using these beads.     

Alkane biodegradation and dynamics of phylogenetic subgroups of sulfate-reducing bacteria in an anoxic coastal marine sediment artificially contaminated with oil

For 503 days, unoiled control and artificially oiled sediments were incubated in situ at 20m water depth in a Mediterranean coastal area. Degradation of the aliphatic fraction of the oil added was followed by GC-MS. At the same time, terminal restriction fragment length polymorphism (T-RFLP) of 16S rRNA encoding genes was used to detect dynamics in the sulfate-reducing bacteria (SRB) community in response to the oil contamination. Specific polymerase chain reaction (PCR) primer sets for five generic or suprageneric groups of SRB were used for PCR amplification of DNA extracted from sediments. During the experiment, hydrocarbons from C(17) to C(30) were significantly degraded even in strictly anoxic sediment layers. Of the five SRB groups, only two groups were detected in the sediments (control and oiled), namely the Desulfococcus-Desulfonema-Desulfosarcina-like group and the Desulfovibrio-Desulfomicrobium-like group. Statistical analysis of community patterns revealed dynamic changes over time within these two groups following the contamination. Significant differences in community patterns were recorded in artificially oiled compared with control sediments. Cloning and sequencing of 16S rRNA encoding genes performed after 503 days showed that many of the most abundant sequences were closely related to hydrocarbonoclastic SRB which could have played an active role in the observed biodegradation of aliphatic hydrocarbons. Results from the present study provide useful information on the dynamics of dominant SRB in heavily oil-contaminated sediments and their potential for anaerobic biodegradation for the treatment of spilled oil in anoxic marine environments.