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Trichloroethylene (TCE) is known as a toxic organic compound found as a pollutant in water streams around the world. The ultimate goal of the present work was to determine the TCE concentration that would be feasible to biodegrade on a long-term basis by a sulfidogenic sludge while maintaining sulfate reducing activity (SRA). Microcosms were prepared with sulfidogenic sludge obtained from a stabilized sulfidogenic UASB and amended with different TCE concentrations (100-300 µM) and two different proportions of volatile fatty acids (VFA) acetate, propionate and butyrate at COD of 2.5:1:1 and 1:1:1, respectively to evaluate the tolerance of the sludge. The overall results suggested that the continuous exposure of the microorganisms to TCE leads to inhibition of SRA; nonetheless, the SRA can be recovered after adequate supplementation of carbon sources and sulfate. The most suitable TCE concentration to operate on a long-term basis while preserving SRA was 26-35 mg L -1 (200-260 µM). A low level of expression of the mRNA of the sulfite reductase subunit alpha (dsrA) gene was obtained in the presence of the TCE and its intermediate products. This gene was associated to SRB belonging to the genera Desulfovibrio, Desulfosalsimonas, Desulfotomaculum, Desulfococcus, Desulfatiglans and Desulfomonas.

Anaerobic technology has a wide scope of application in different areas such as manufacturing, food industry, and agriculture. Nowadays, it is mainly used to produce electrical and thermal energy from crop processing, solid waste treatment or wastewater treatment. More intensively, trend nowadays is usage of this technology biodegradable and biomass waste processing and biomethane or hydrogen production. In this paper, the diversities of sulfate-reducing bacteria (SRB) under different imputed raw material to the bioreactors were characterized. These diversities at the beginning of sampling and after cultivation were compared. Desulfovibrio, Desulfobulbus, and Desulfomicrobium genus as dominant among sulfate reducers in the bioreactors were detected. The Desulfobulbus species were dominant among other SRB genera before cultivation, but these bacteria were detected only in three out of the seven bioreactors after cultivation dominant.

The kinetics and microbial ecology in sulfidogenic bioreactors used in a novel two-stage process for co-treatment of acid mine drainage (AMD) and municipal wastewater (MWW) were investigated. Michaelis-Menten modeling of COD oxidation by sulfate reducing bacteria (SRB) (Vmax=0.33mgL(-1)min(-1), Km=4.3mgL(-1)) suggested that the Vmax can be reasonably achieved given the typical COD values in MWW and anticipated mixing with AMD. Non-competitive inhibition modeling (Ki=6.55mgL(-1)) indicated that excessive iron level should be avoided to limit its effects on SRB. The COD oxidation rate was positively correlated to COD/sulfate ratio and SRB population, as evidenced by dsrA gene copies. Phylogenetic analysis revealed diverse microbial communities dominated by sulfate reducing delta-proteobacteria. Microbial community and relative quantities of SRB showed significant differences under different COD/sulfate ratios (0.2, 1 and 2), and the highest dsrA gene concentration and most complex microbial diversity were observed under COD/sulfate ratio 2. Major species were associated with Desulfovirga, Desulfobulbus, Desulfovibrio, and Syntrophus sp. The reported COD kinetics, SRB abundances and the phylogenetic profile provide insights into the co-treatment process and help identify the parameters of concerns for such technology development., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study evaluated the effects of operational parameters and type of substrate on the abundance of sulphate-reducing bacteria in 25 industrial biogas digesters using qPCR targeting the functional dissimilatory sulphite reductase gene. The aim was to find clues for operational strategies minimizing the production of H2S. The results showed that the operation, considering strategies evaluated, only had scarce effect on the abundance, varying between 10(5) and 10(7) gene copies per ml. However, high ammonia levels and increasing concentration of sulphate resulted in significantly lower and higher levels of sulphate-reducing bacteria, respectively., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Injection of up-flow packed-bed bioreactors with excess volatile fatty acids and limiting concentrations of nitrate and sulfate gave complete reduction of nitrate from 0 to 5.5 cm and complete or near-complete reduction of sulfate from 3.2 to 11.5 cm along the bioreactor flow path. Most of the biomass (85%) and most of the genes for nitrate reduction (narG, 96%; napA, 99%) and for sulfate reduction (dsrB, 91%) were present near the inlet (0-5.5 cm) of the 37-cm-long bioreactor. Microbial community analysis by a combination of denaturing gradient gel electrophoresis and pyrosequencing of 16S rRNA amplicons indicated that nitrate-reducing Arcobacter and Pseudomonas species were located from 0 to 3.2 cm and throughout, respectively. Desulfobulbus species were the main sulfate reducers present and acetotrophic methanogens of the genus Methanosaeta predominated at 20-37 cm. Overall, the results indicated a succession of microbial communities along the bioreactor flow path. In the absence of nitrate, the sulfate reduction zone moved nearer to the bioreactor inlet. The sulfide concentration in the bioreactor effluent was temporarily lowered after nitrate injection was re-started. Hence, the bioreactor sulfide output could be disrupted by pulsed, not by constant nitrate injection, as demonstrated also previously in a low-temperature oil field.

Sulfate-reducing bioreactors are a promising option for the treatment of acid mine drainage. We studied the structure and function of a biofilm in a methanol-fed fixed-bed in-lake reactor for the treatment of an acidic pit lake by a combination of laboratory incubations, chemical and molecular analyses and confocal laser scanning microscopy to determine whether competition by different groups of microorganisms as well as the precipitation of minerals affect reactor performance negatively. The biofilm growing on the surface of a synthetic carrier material consisted of dense microbial colonies covered by iron-sulfide precipitates. The microorganisms continuously had to overgrow this mineral coating, resulting in a high biomass turnover. About one third of the added methanol was used by sulfate reduction, and the rest by competing reactions. Sulfate-reducing bacteria as well as methanogens and acetogens were involved in methanol consumption. Six different groups of Deltaproteobacteria, dominated by the genera Desulfomonile, Desulfobacterium and a phylotype related to Geobacter, Gram-positive sulfate reducers of the genus Desulfosporosinus, acetogenic Acetobacteria, different fermenting bacteria as well as methylotrophic methanogens were identified. The versatility of the microbial food web is probably an important factor stabilizing the biofilm function under fluctuating and partly oxidizing conditions in the reactor.

High rate sulfate reduction under acidic conditions opens possibilities for new process flow sheets that allow the selective recovery of metals from mining and metallurgical waste and process water. However, knowledge about high-rate sulfate reduction under acidic conditions is limited. This paper investigates sulfate reduction in a membrane bioreactor at a controlled pH of 5. Sulfate and formate were dosed using a pH-auxostat system while formate was converted into hydrogen, which was used for sulfate reduction. Sulfide was removed from the gas phase to prevent sulfide inhibition. This study shows a high-rate sulfate-reducing bioreactor system for the first time at pH 5, with a volumetric activity of 188mmol SO(4)(2-)/I/d and a specific activity of 81mmol SO(4)(2-) volatile suspended. The microbial community at the end of the reactor run consisted of a diverse mixed population including sulfate-reducing bacteria.

The characteristics of the biofilm and the solids formed during the operation of a sulphate-reducing fixed-bed reactor, fed with a moderately acidic synthetic effluent containing zinc and iron, are presented. A diverse population of delta-Proteobacteria SRB, affiliated to four distinct genera, colonized the system. The morphology, mineralogy and surface chemistry of the precipitates were studied by X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive X-ray (EDX). The XRD patterns observed are characteristic of amorphous solid phases. Peaks corresponding to crystalline iron sulphide, marcasite, sphalerite and wurtzite were also identified. SEM-EDX results confirm the predominance of amorphous phases appearing as a cloudy haze. EDX spectra of spots on the surface of these amorphous phases reveal the predominance of iron, zinc and sulphur indicating the formation of iron and zinc sulphides. The predominance of these amorphous phases and the formation of very fine particles, during the operation of the SRB column, are in agreement and can be explained by the formation pathways of metal sulphides at ambient temperature, alkaline pH and reducing conditions. Solids are precipitated either as (i) amorphous phases deposited on the bed material, as well as on surface of crystals, e.g. Mg(3)(PO(4))(2) and (ii) as rod-shaped solids characterized by a rough hazy surface, indicating the encapsulation of bacterial cells by amorphous metal sulphides.

The microbial population structure and function of natural anaerobic communities maintained in lab-scale continuously stirred tank reactors at different lactate to sulfate ratios and in the absence of sulfate were analyzed using an integrated approach of molecular techniques and chemical analysis. The population structure, determined by denaturing gradient gel electrophoresis and by the use of oligonucleotide probes, was linked to the functional changes in the reactors. At the influent lactate to sulfate molar ratio of 0.35 mol mol(-1), i.e., electron donor limitation, lactate oxidation was mainly carried out by incompletely oxidizing sulfate-reducing bacteria, which formed 80-85% of the total bacterial population. Desulfomicrobium- and Desulfovibrio-like species were the most abundant sulfate-reducing bacteria. Acetogens and methanogenic Archaea were mostly outcompeted, although less than 2% of an acetogenic population could still be observed at this limiting concentration of lactate. In the near absence of sulfate (i.e., at very high lactate/sulfate ratio), acetogens and methanogenic Archaea were the dominant microbial communities. Acetogenic bacteria represented by Dendrosporobacter quercicolus-like species formed more than 70% of the population, while methanogenic bacteria related to uncultured Archaea comprising about 10-15% of the microbial community. At an influent lactate to sulfate molar ratio of 2 mol mol(-1), i.e., under sulfate-limiting conditions, a different metabolic route was followed by the mixed anaerobic community. Apparently, lactate was fermented to acetate and propionate, while the majority of sulfidogenesis and methanogenesis were dependent on these fermentation products. This was consistent with the presence of significant levels (40-45% of total bacteria) of D. quercicolus-like heteroacetogens and a corresponding increase of propionate-oxidizing Desulfobulbus-like sulfate-reducing bacteria (20% of the total bacteria). Methanogenic Archaea accounted for 10% of the total microbial community.

A novel sulfate-reducing bacterium, strain HB1(T), was isolated from an upflow anaerobic sludge blanket (UASB) reactor treating paper-mill wastewater operated at 37 degrees C. Cells of strain HB1(T) were oval to rod-shaped, 1-1.3 microm wide and 2.6-3.5 microm long and Gram-negative. The optimum temperature for growth was 28-30 degrees C. In the presence of sulfate, the isolate was able to grow on H(2)/acetate, formate, ethanol, propionate, fumarate, succinate, butyrate, crotonate, catechol, benzoate, 4-hydroxybenzoate, palmitate and stearate. The isolate only grew on H(2) when acetate was added as a carbon source; when grown on formate, acetate was not required. Growth was also possible on pyruvate and crotonate without an electron acceptor. The isolate showed very poor growth on acetate. Thiosulfate and sulfate were used as electron acceptors. Phylogenetic analysis of 16S rRNA gene sequences revealed that strain HB1(T) represents a novel lineage within the Deltaproteobacteria; sequence similarities between strain HB1(T) and members of other related genera were less than 91%. Strain HB1(T) was also distinguished from members of related genera based on differences in several phenotypic characteristics. It is a member of the family Desulfobacteraceae. The major cellular fatty acids of strain HB1(T) were C(16:0), iso-C(15:0), anteiso-C(15:0) and C(14:0). beta-Hydroxy fatty acids were also present in the range of C(14:0) to C(18:0), of which C(16:0) was the most abundant. The G+C content of the DNA was 55.1 mol%. Based on physiological, biochemical and chemotaxonomic traits together with results of comparative 16S rRNA gene sequence analysis, strain HB1(T) is considered to represent a novel species in a new genus, for which the name Desulfatirhabdium butyrativorans gen. nov., sp. nov. is proposed. The type strain of Desulfatirhabdium butyrativorans is HB1(T) (=DSM 18734(T) =JCM 14470(T)).

Phosphorus removal and denitrification using iron and wood as electron donors were examined in a laboratory-scale biological filter reactor. Phosphorus removal and denitrification using iron and wood continued for 1,200 days of operation. Wood degradation by heterotrophic denitrification and iron oxidation by hydrogenotrophic denitrification occurred simultaneously. In the biofilm inside the wood, not only heterotrophic denitrification activity but also sulfate reduction and sulfur denitrification activities were recognized inside the wood, indicating that a sulfur oxidation-reduction cycle was established. Sulfate reduction and denitrification were accelerated with the addition of cellulose. Microbial communities of sulfate-reducing bacteria by PCR primer sets could be amplified in the biofilm in the reactors. The dissimilatory sulfite reductase gene and the 16S rRNA gene of six phylogenetic groups of SRB in the reactors were analyzed. Some SRB group-specific primers-amplification products were obtained inside the wood and around iron., (Copyright IWA Publishing 2008.)

Acetate utilization in sulfidogenic fluidized-bed reactors (FBRs) was investigated for the treatment of iron containing wastewater at low (8 degrees C) and high (65 degrees C) temperatures. The FBRs operated at low and high temperatures were inoculated with cultures of sulfate-reducing bacteria (SRB) originally enriched from arctic and hot mining environments, respectively. Acetate was not utilized as a carbon and electron source for SRB at 8 degrees C. With ethanol, hydrogen sulfide was produced from ethanol to acetate oxidation, which precipitated the iron. Then, several attempts were made to obtain acetate oxidation at 8 degrees C. Inoculation of two different low temperature enrichments and operating the FBR for a long period of time (321 days) did not result in enrichment of acetate oxidizing SRB. Due to the absence of acetate oxidation at 8 degrees C, external alkalinity addition was required to keep the pH neutral. At 65 degrees C, average acetate and sulfate removals were 52+/-12% and 24+/-8% at 670 mg/Ld acetate and 1500 mg/Ld sulfate loadings, respectively. The produced alkalinity from acetate oxidation increased the pH from 6.4 to around 7.5 and electron flow to sulfate reduction averaged 65%. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes showed quite stable SRB community at 8 degrees C, whereas, at 65 degrees C SRB community was dynamic. In the FBRs, Desulfomicrobium apsheronum and Desulfosporosinus sp. at 8 degrees C and Desulfotomaculum sp. at 65 degrees C were detected.

Three anaerobic, dechlorinating consortia were enriched from different sites using methanol and tetrachloroethene (PCE) and maintained for approximately 3 years. These consortia were evaluated using chemical species analysis including distribution of dechlorination products, production of organic acids and methane, and using qualitative and quantitative PCR (qPCR), terminal restriction fragment length polymorphism (TRFLP), and denaturing gradient gel electrophoresis (DGGE) with primers specific to Dehalococcoides 16S rRNA gene sequences. TRFLP and analysis of organic acids revealed differing fermentative populations in each consortium, which were dominated by acetogens. Monitoring methane production combined with qPCR for archaea showed that complete dechlorination of PCE-to-ethene occurred in the presence and absence of methanogens. The 16S rRNA gene-based analyses demonstrated that enrichment with PCE resulted in dechlorinating communities dominated by Dehalococcoides and Dehalobacter, and that up to four different PCE-dechlorinating organisms coexisted in one consortium. Further, the DGGE analysis suggested that at least one consortium contained multiple Dehalococcoides strains. The combined analysis of 16S rRNA and reductive dehalogenase genes suggested that one consortium contained a member of the Dehalococcoides "Cornell" group with the ability to respire VC.

The diversity and the community structure of sulfate-reducing bacteria (SRB) in an anaerobic continuous bioreactor used for treatment of a sulfate-containing wastewater were investigated by fluorescence in situ hybridization. Hybridization to the 16S rRNA probe EUB338 for the domain Bacteria was performed, followed by a nonsense probe NON338 as a control for nonspecific staining. Sulfate-reducing consortia were identified by using five nominally genus-specific probes (SRB129 for Desulfobacter, SRB221 for Desulfobacterium, SRB228 for Desulfotomaculum, SRB660 for Desulfobulbus, and SRB657 for Desulfonema) and four group-specific probes (SRB385 as a general SRB probe, SRB687 for Desulfovibrioaceae, SRB814 for Desulfococcus group, and SRB804 for Desulfobacteriaceae). The total prokaryotic population was determined by 4',6-diamidino-2-phenylindole staining. Hybridization analysis using these 16S rRNA-targeted oligonucleotide probes showed that, of those microbial groupings investigated, Desulfonema, Desulfobulbus, spp., and Desulfobacteriaceae group were the main sulfate-reducing bacteria in the bioreactor when operated at steady state at 35 degrees C, pH 7.8, and a 2.5-day residence time with feed stream containing 2.5 kg m-3 sulfate as terminal electron acceptor and 2.3 kg m-3 acetate as carbon source and electron donor.

Here we describe the diversity and activity of sulfate-reducing bacteria (SRB) in sulfidogenic bioreactors by using the simultaneous analysis of PCR products obtained from DNA and RNA of the 16S rRNA and dissimilatory sulfite reductase (dsrAB) genes. We subsequently analyzed the amplified gene fragments by using denaturing gradient gel electrophoresis (DGGE). We observed fewer bands in the RNA-based DGGE profiles than in the DNA-based profiles, indicating marked differences in the populations present and in those that were metabolically active at the time of sampling. Comparative sequence analyses of the bands obtained from rRNA and dsrB DGGE profiles were congruent, revealing the same SRB populations. Bioreactors that received either ethanol or isopropanol as an energy source showed the presence of SRB affiliated with Desulfobulbus rhabdoformis and/or Desulfovibrio sulfodismutans, as well as SRB related to the acetate-oxidizing Desulfobacca acetoxidans. The reactor that received wastewater containing a diverse mixture of organic compounds showed the presence of nutritionally versatile SRB affiliated with Desulfosarcina variabilis and another acetate-oxidizing SRB, affiliated with Desulfoarculus baarsii. In addition to DGGE analysis, we performed whole-cell hybridization with fluorescently labeled oligonucleotide probes to estimate the relative abundances of the dominant sulfate-reducing bacterial populations. Desulfobacca acetoxidans-like populations were most dominant (50 to 60%) relative to the total SRB communities, followed by Desulfovibrio-like populations (30 to 40%), and Desulfobulbus-like populations (15 to 20%). This study is the first to identify metabolically active SRB in sulfidogenic bioreactors by using the functional gene dsrAB as a molecular marker. The same approach can also be used to infer the ecological role of coexisting SRB in other habitats.

The microbial population structure and function of a mixed culture of sulfate-reducing bacteria (SRB) maintained in anaerobic continuous bioreactors were tracked before and after a major perturbation, which involved the addition of sulfate to the influent of a bioreactor when operated at steady state at 35 degrees C, pH 7.8 and a 2.5 day residence time with feed stream containing 10 and 15 kg m(-3) sulfate as terminal electron acceptor and 19.6 and 29.4 kg m(-3) ethanol as carbon source and electron donor, respectively. The population structure determined by fluorescence in situ hybridization (FISH), by using 16S rRNA-targeted oligonucleotide probes, was linked to the functional performance of the SRB in the reactor. Hybridization analysis using these 16S rRNA-targeted oligonucleotide probes revealed that a high concentration of sulfate was toxic for Desulfobacterium and Desulfobulbus. On the other hand, the Desulfococcus group was found to be the most dominant group of SRB in the feed stream containing 15 kg m(-3) sulfate as terminal electron acceptor and 29.4 kg m(-3) ethanol as carbon source and electron donor.

The objective of this research is to make a novel wastewater treatment process activated by a sulfur-redox cycle action of microbes in low temperature conditions. This action is carried out by sulfate-reducing bacteria (SRB) and sulfur-oxidizing bacteria (SOB). The process was comprised of a UASB reactor as pre-treatment and an aerobic downflow hanging sponge (DHS) reactor as post-treatment. As the results of reactor operation, the whole process achieved that over 90% of CODcr removal efficiency, less than 30 mgCODcr/L (less than 15 mgBOD/L) of final effluent, at 12 h of HRT and at 8 degrees C of UASB reactor temperature. Acetobacterium sp. was detected as the predominant species by PCR-DGGE method targeting 16SrDNA with band excision and sequence analysis. In the UASB reactor, various species of sulfate-reducing bacterium, Desulfobulbus sp., Desulfovibrio sp., and Desulfomicrobium sp., were found by cloning analysis. In the DHS reactor, Tetracoccus sp. presented as dominant. The proposed sulfur-redox action process was considered as an applicable process for low strength wastewater treatment in low temperature conditions.

Analyses of microbial community structure and the relationships between Sulfate-Reducing Bacteria (SRBs) and Acidogenic Bacteria (ABs) in a completely stirred sulfate-reducing reactor were carried out by modified polymerase chain reaction-single-stranded conformation polymorphism (PCR-SSCP) targeted eubacterial 16S ribosomal RNA gene. A total of 13 bands were obtained and 6 of them (A1, A3, A4, A5, A9, A10) were sequenced. The sequences are similar to Leuconostoc mesenteroides (GenBank Access No. AY453065), some uncultured bacteria (AJ318147, AF227834, AJ576427), Ethanologenbacterium (AY434722), Clostridiaceae (AB084627), etc. In order to investigate the SRBs in the reactor,the active sludge was cultured on SRB-selected media and also did SSCP with the compound cultured bacteria. Two new bands appeared, one similar to Bacteroidetes (AB074606) and another similar to Desulfovibrio (Y12254, U42221). The experimental results indicate that the proportion of SRBs in the reactor is probably less than 1.5 percent. But the few SRBs play a very important role during the course of sulfate reduction via cooperation with acidogenic

The consumption of methane in anoxic marine sediments is a biogeochemical phenomenon mediated by two archaeal groups (ANME-1 and ANME-2) that exist syntrophically with sulfate-reducing bacteria. These anaerobic methanotrophs have yet to be recovered in pure culture, and key aspects of their ecology and physiology remain poorly understood. To characterize the growth and physiology of these anaerobic methanotrophs and the syntrophic sulfate-reducing bacteria, we incubated marine sediments using an anoxic, continuous-flow bioreactor during two experiments at different advective porewater flow rates. We examined the growth kinetics of anaerobic methanotrophs and Desulfosarcina-like sulfate-reducing bacteria using quantitative PCR as a proxy for cell counts, and measured methane oxidation rates using membrane-inlet mass spectrometry. Our data show that the specific growth rates of ANME-1 and ANME-2 archaea differed in response to porewater flow rates. ANME-2 methanotrophs had the highest rates in lower-flow regimes (mu(ANME-2) = 0.167 . week(-1)), whereas ANME-1 methanotrophs had the highest rates in higher-flow regimes (mu(ANME-1) = 0.218 . week(-1)). In both incubations, Desulfosarcina-like sulfate-reducing bacterial growth rates were approximately 0.3 . week(-1), and their growth dynamics suggested that sulfate-reducing bacterial growth might be facilitated by, but not dependent upon, an established anaerobic methanotrophic population. ANME-1 growth rates corroborate field observations that ANME-1 archaea flourish in higher-flow regimes. Our growth and methane oxidation rates jointly demonstrate that anaerobic methanotrophs are capable of attaining substantial growth over a range of environmental conditions used in these experiments, including relatively low methane partial pressures.

Here, we describe a three-step nested-PCR-denaturing gradient gel electrophoresis (DGGE) strategy to detect sulfate-reducing bacteria (SRB) in complex microbial communities from industrial bioreactors. In the first step, the nearly complete 16S rRNA gene was amplified using bacterial primers. Subsequently, this product was used as a template in a second PCR with group-specific SRB primers. A third round of amplification was conducted to obtain fragments suitable for DGGE. The largest number of bands was observed in DGGE patterns of products obtained with primers specific for the Desulfovibrio-Desulfomicrobium group, indicating a large diversity of these SRBs. In addition, members of other phylogenetic SRB groups, i.e., Desulfotomaculum, Desulfobulbus, and Desulfococcus-Desulfonema-Desulfosarcina, were detected. Bands corresponding to Desulfobacterium and Desulfobacter were not detected in the bioreactor samples. Comparative sequence analysis of excised DGGE bands revealed the identity of the community members. The developed three-step PCR-DGGE strategy is a welcome tool for studying the diversity of sulfate-reducing bacteria.

The identification of bacteria in oil production facilities has previously been based on culture techniques. However, cultivation of bacteria from these often-extreme environments can lead to errors in identifying the microbial community members. In this study, molecular techniques including fluorescence in situ hybridization, PCR, denaturing gradient gel electrophoresis, and sequencing were used to track changes in bacterial biofilm populations treated with nitrate, nitrite, or nitrate+molybdate as agents for the control of sulfide production. Results indicated that nitrite and nitrate+molybdate reduced sulfide production, while nitrate alone had no effect on sulfide generation. No long-term effect on sulfide production was observed. Initial sulfate-reducing bacterial numbers were not influenced by the chemical treatments, although a significant increase in sulfate-reducing bacteria was observed after termination of the treatments. Molecular analysis showed a diverse bacterial population, but no major shifts in the population due to treatment effects were observed.