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Little is known about the microbial ecology of closed landfill, or how the diversity of these communities changes over time. This study focuses on the bacterial community diversity analysis of landfill leachate (LFL) sourced from Powerstown landfill Co. Carlow, Ireland. The LFL samples used in this study came from both closed and active landfill, and represent different phases of the landfill. HiSeq sequencing systems based 16S rRNA gene sequence analysis was used to investigate the bacterial communities of a total of six LFL samples; four samples from landfill cells (two from closed landfill cells and two from active landfill cells), and two samples sourced from a leachate tank (LT) collected over an interval of 2.5 years. Members of the phylum Proteobacteria (Pseudomonadota) were dominant in all landfill samples investigated (56 to 71%), followed by Bacteroidetes (Bacteroidota) (7 to 21%). The operating status of landfills (i.e. active / closed) and the age of landfill were found to affect the bacterial composition of LFL. Members of the class Epsilonproteobacteria and genus Sulfurimonas were dominant in LFL from active landfill, while the class Betaproteobacteria and genus Pusillimonas were dominant in closed landfill. Furthermore, it was observed that the composition of the microbial community in active landfill showed fluctuation by the progressing landfill age, while the bacterial composition of closed landfill tends to be stable as the year the landfill is closed.

Renewable energy has become increasingly popular in the recent times as countries have shifted focus towards carbon-neutral and environmentally sustainable sources of energy. Biogas and biomethane derived energy from Anaerobic Digestion of organic waste have proved to be excellent alternative to fossil-fuel based energy. Many countries around the world are actively adopting biomethane as an alternative to petrol/diesel or use biogas to provide energy for the domestic sector through combined heat and power plants. The contribution of anaerobic digestion in the Irish energy sector remains quite low as compared to its European union counterparts due to related legislation and lack of government incentives. Nevertheless, Ireland is rich in biomass with a huge potential for developing its renewable energy sector from anaerobic digestion of agricultural waste, which is relatively inexpensive and available here in abundance. Chicken litter is one such waste which offers a variety of environmental benefits as anaerobic digestion feedstock. In comparison to cow manure and pig manure, chicken litter was relatively unpopular in anaerobic digestion due to its high nitrogen and lignocellulose content. Further, the global production of chicken manure has been increasing constantly thus making its disposal challenging across the world. However, researchers have identified many optimisation strategies to overcome these issues and demonstrated the advantages of using chicken litter in anaerobic digestion including odor reduction, greenhouse gas mitigation, production of gaseous biofuel, avoiding eutrophication of water bodies and significantly improving global production of renewable energy to help countries reduce their carbon emissions.

Landfilling is one of the most common and widely accepted practices for the disposal of waste throughout the world. Leachate, a major drawback of landfilling, continues to be produced at vast rates and current treatment options are costly and often inadequate. The management of leachate is of economic and environmental importance, due to its potential to cause contamination to ground and surface water. This research focuses on treating leachate in a cost-effective manner through bioremediation. Microorganisms were isolated from landfill leachate (LFL) and screened to determine their ability to remediate a wide range of compounds found in leachate, such as ammonia, phosphate and nitrate. Selected isolates were identified as belonging to the phylum’s Firmicutes, Actinobacteria, and Proteobacteria, isolates were inoculated into soil contained in a fixed bed column system. The column system was optimised and used for the treatment of LFL over a 10-hour period. High percentage removal rates were achieved for ammonia (>90%) and removal nitrate and phosphate (>60%). Although EPA discharge limits were not achieved, bioremediation using selected microbial strains represents a cost effective treatment option when compared to conventional methods. Research is now required to further optimise this system to achieve discharge limits for all compounds tested.

Biomethane production potential was evaluated by anaerobic co-digestion of grass silage (GS) and chicken litter (CL) in batch experiments. The aim of this study was to identify optimum enzyme treatment and co-digestion ratio for these substrates. The study is divided into trial A and trial B for enzyme treatment and co-digestion respectively. Mixed substrates were utilised in both trials which were maintained for a period of 30 days in 125 mL serum bottles at 37 °C. Preliminary batch assays in trial A for biomethane potential determination were performed using a co-digestion ratio of 2:1 (GS:CL) treated with five different concentrations of enzyme (w/v)—0.05, 0.1, 0.25, 0.5 and 1%. The highest specific methane yield (SMY) of 61 L CH4/kg was observed for 0.1% (w/v) enzyme treatment. Trial B assays were conducted for nine different GS:CL ratios—0:1, 1:0, 1:1, 1:2, 1:3, 2:3, 3:2, 3:1, and 2:1 respectively. Highest SMY were recorded for the ratios 1:0, 1:1 and 1:3 of GS:CL which correspond to 124 L CH4/kg, 137 L CH4/kg and 135 L CH4/kg respectively.

Due to the extensive use of organotin compounds (OTCs) in numerous human activities, large amounts have been introduced to various ecosystems. Several appear on the priority pollutant lists of the ...

Tributyltin (TBT) is a synthetic organotin compound and is recognised as the most toxic chemical ever knowingly introduced into the marine environment. Organotins have a wide range of application with an estimated global production of 50,000 tons per year therefore impacts from TBT can be seen or anticipated in many environments including protected areas like aqua farms and world heritage sites effecting aquaculture and undisturbed ecosystems. Due to the impact of TBT a necessity exists for the treatment of contaminated sediment and soil using nature-based solutions as it is both non-invasive and sustainable. However these remediation techniques must also be economically interesting to increase its acceptance. Thus using the biomass from plant species with the ability to cultivate in dredged sediment for biofuel and biomaterials production should be considered to ensure that renewable energy consumption of 10% is meet by 2020. The objectives of this study is to investigate the potential of Hordeum vulgare and Brassica napus to cultivate in marine sediments despite the salinity and TBT pollution for a 30 day pot trial. Further to inoculate seedling roots with known TBT degrading bacteria to enhance the phytoremediation capacity. Also to establish if bacterial isolates C7 and C22 are plant growth promoting bacteria. Statistical analysis on seedlings biomass was carried out followed by analytical analyses by atomic absorption spectroscopy (AAS) and sediment analysis by gas chromatography mass spectrometry (GC-MS). Results showed a maximum decrease of ≥ 82% and ≥ 80% of TBT in sediment samples respectively with bacteria inoculum C7 (KX881905) and C22 (KX881904), further Hordeum vulgare seedlings were the ideal candidate plant species with a significant production of biomass.

Four low-cost materials, oyster shells, pumice stone, sand and zeolite were employed as adsorbents in an adsorption batch assays investigating the removal of ammonia, phosphate and nitrate from an aqueous solution. These compounds were chosen as they represent typical compounds found in landfill leachate (LFL). Assay performance was evaluated by the Langmuir and Freundlich adsorption isotherms. The top two materials, oyster shells and pumice stone, were employed as adsorbents in a fixed-bed column trial examining the effect of bed height and flow rate on the treatment of a synthetic LFL. The trial concluded that the highest rates of adsorption were achieved using bed heights of 20 cm with a flow rate of 5 mL min

Anaerobic digestion (AD) of chicken litter (CL) is a viable alternative to disposal. However, methane yields from this primarily organic waste are quite low when mono-digested. This paper discusses the effect of an enzyme cocktail, trace element (TE) supplementation and selenium (Se) addition in small-scale batch biomethane potential (BMP) assays to enhance the AD of CL. Eleven different assays were set up in triplicate including assays containing only inoculum (blank), only CL (negative control) and cellulose and inoculum (positive control). The results indicate that both enzyme treatment and trace element supplementation enhanced the biogas and methane yield. The highest specific biogas and methane yields were noted for 1% enzyme-treated CL of 835.2 L/kg volatile solids (VS) and 460.8 L/kg VS, respectively. Usually, mono-digestion of CL is low due to high nitrogen content and the presence of recalcitrant lignocellulosic material from the bedding material. Enzyme treatment performed better than the addition of the TE mix and Se.

The use of phytoremediation to sustainably recover areas contaminated by toxic heavy metals such as cadmium (Cd) has been made feasible since the discovery of hyperaccumulator plants. This study examines the potential of the invasive Impatiens glandulifera for phytoremediation propensity of Cd. In these experiments, the plants were exposed to and tested for Cd accumulation, the propensity to accumulate other heavy metals, such as Zinc, was not investigated. The efficacy of phytoaccumulation was assessed over two trials (Cd concentrations of 20 mg/kg to 150 mg/kg) via examination of bioconcentration factor (BCF), translocation factor (TF), and total removal (TR). Exposure to Cd levels of up to 150 mg/kg in the trials did not affect the biomass of the plants compared to the control. Impatiens glandulifera accumulated cadmium at a rate of 276 to 1562 mg/kgin stems, with BCFs, TFs, and TRs of 64.6 to 236.4, 0.2 to 1.2, and 3.6 to 29.2 mg Cd, respectively. In vitro germination revealed unprecedented germination ability, demonstrating the remarkable hypertolerance of I. glandulifera, with no significant difference in the germination of seedlings exposed to 1000 mg/kg Cd compared to the control. This study also examined the localization of Cd in plant tissues via a histochemical assay using dithizone. The results presented herein suggest that I. glandulifera can act as a hyperaccumulator of Cd for phytoremediation.

Tributyltin (TBT) is one of the most toxic anthropogenic compounds introduced into the marine environment. Despite its global ban in 2008, TBT is still a problem of great concern due to its high affinity for particulate matter, providing a direct and potentially persistent route of entry into benthic sediments. Bioremediation strategies may constitute an alternative approach to conventional physicochemical methods, benefiting from the microorganism’s potential to metabolize anthropogenic compounds. In this work, a simple, precise and accurate static headspace gas chromatography method was developed to investigate the ability of TBT degrading microbes in sedimentary microcosms over a period of 120 days. The proposed method was validated for linearity, repeatability, accuracy, specificity, limit of detection and limit of quantification. The method was subsequently successfully applied for the detection and quantification of TBT and degrading compounds in sediment samples on day 0, 30, 60, 90 and 120 of the experiment employing the principles of green chemistry. On day 120 the concentration of TBT remaining in the microcosms ranged between 91.91 ng/g wet wt for the least effective microbial inoculant to 52.73 ng/g wet wt for the most effective microbial inoculant from a stating concentration of 100 ng/g wet wt.

BACKGROUND Metal sulfide recovery in sulfate reducing bioreactors is a challenge due to the formation of small precipitates with poor settling properties. The size of the metal sulfide precipitates with the change in operational parameters such as pH, sulfide concentration and reactor configuration has been previously studied. The effect of the hydraulic retention time (HRT) on the metal precipitate characteristics such as particle size for settling has not yet been addressed. RESULTS The change in size of the metal (Cu, Zn, Pb and Cd) sulfide precipitates as a function of the HRT was studied in two sulfate reducing inversed fluidized bed (IFB) reactors operating at different chemical oxygen demand concentrations to produce high and low sulfide concentrations. The decrease of the HRT from 24 to 9 h in both IFB reactors affected the contact time of the precipitates formed, thus making differences in aggregation and particle growth regardless of the differences in sulfide concentration. Further HRT decrease to 4.5 h affected the sulfate reducing activity for sulfide production and hence, the supersaturation level and solid phase speciation. Metal sulfide precipitates affected the sulfate reducing activity and community in the biofilm, probably because of the stronger local supersaturation causing metal sulfides accumulation in the biofilm. CONCLUSIONS This study shows that the HRT is an important factor determining the size and thus the settling rate of the metal sulfides formed in bioreactors. © 2013 Society of Chemical Industry

Aims: The objective of this work was to provide functional evidence of key metabolic pathways important for anaerobic digestion processes through the identification of highly expressed proteins in a mixed anaerobic microbial consortium. Methods and Results: The microbial communities from an anaerobic industrial-like wastewater treatment bioreactor were characterized using phylogenetic analyses and metaproteomics. Clone libraries indicated that the bacterial community in the bioreactor was diverse while the archaeal population was mainly composed of Methanocorpusculum-like (76%) micro-organisms. Three hundred and eighty-eight reproducible protein spots were obtained on 2-D gels, of which 70 were excised and 33 were identified. The putative functions of the proteins detected in the anaerobic bioreactor were related to cellular processes, including methanogenesis from CO2 and acetate, glycolysis and the pentose phosphate pathway. Metaproteomics also indicated, by protein assignment, the presence of specific micro-organisms in the bioreactor. However, only a limited overlap was observed between the phylogenetic and metaproteomic analyses. Conclusions: This study provides some direct evidence of the microbial activities taking place during anaerobic digestion. Significance and Impact of Study: This study demonstrates metaproteomics as a useful tool to uncover key biochemical pathways underpinning specific anaerobic bioprocesses.

The reproducibility and stability of low- temperature anaerobic wastewater treatment systems undergoing transient perturbations was investigated. Three identical anaerobic expanded granular sludge bed-based bioreactors were used to degrade a volatile fatty acid and glucose-based wastewater under sub-ambient (15 degrees C) conditions. The effect of a variety of environmental perturbations on bioreactor performance was assessed by chemical oxygen demand removal. Temporal microbial community development was monitored by denaturation gradient gel electrophoresis (DGGE) of 16S rRNA genes extracted from sludge granules. Methanogenic activity was monitored using specific methanogenic activity assays. Bioreactor performance and microbial population dynamics were each well replicated between both experimental bioreactors and the control bioreactor prior to, and after the implementation of most of the applied perturbations. Gene fingerprinting data indicated that Methanosaeta sp. were the persistent, keystone members of the archaeal community, and likely were pivotal for the physical stability and maintenance of the granular biofilms. Cluster analyses of DGGE data suggested that temporal shifts in microbial community structure were predominantly independent of the applied perturbations.

Two laboratory-scale expanded granular sludge bed (EGSB) anaerobic bioreactors (R1 and R2) were inoculated with biomass from different mesophilic (37 degrees C) treatment plants, and used for the treatment of an organic solvent-based wastewater at 9-14 degrees C at applied organic loading rates (OLRs) of 1.2-3.6kg chemical oxygen demand (COD)m(-3)d(-1). Replicated treatment performance was observed at 10-14 degrees C, which suggested the feasibility of the process at pilot-scale. Stable and efficient COD removal, along with high methane productivity, was demonstrated at 9 degrees C at an applied OLR of 2.4kgCODm(-3)d(-1). Clonal libraries and fluorescence in situ hybridization (FISH) indicated that the seed sludges were dominated (60%) by acetoclastic Methanosaeta-like organisms. Specific methanogenic activity (SMA) profiles indicated shifts in the physiological profiles of R1 and R2 biomass, including the development of psychrotolerant methanogenic activity. Acetoclastic methanogenesis represented the primary route of methane production in R1 and R2, which is in contrast with several previous reports from low-temperature bioreactor trials. A reduction in the abundance of Methanosaeta-like clones (R2), along with the detection of hydrogenotrophic methanogenic species, coincided with altered granule (sludge) morphology and the development of hydrogenotrophic SMA after prolonged operation at 9 degrees C.

Radiotracer incubation experiments and beta microimaging, along with fluorescent in situ hybridisation (FISH), are proposed as a complementary approach to specific methanogenic activity testing and measurement of in vitro substrate utilisation rates to understand better the ecophysiology of anaerobic granular biofilms from wastewater treatment reactors.

Thirteen anaerobic hybrid expanded granular sludge bed-anaerobic filter bioreactors were used for psychrophilic (15–18 °C) anaerobic digestion of a variety of synthetic and non-synthetic wastewaters, including: food-processing, dairy, aromatic- and aliphatic-containing and brewery discharges. Specific methanogenic activity assays were employed to assess temporal physiological activity dynamics. Terminal restriction fragment length polymorphism genetic fingerprinting and fluorescent in situ hybridization were used to monitor shifts in the structure of the microbial communities in the bioreactors in response to operating conditions. Treatment efficiencies obtained were comparable to previous mesophilic (37 °C) trials. Methanogenic activity developed under psychrophilic conditions and putative psychrophilic populations were detected within otherwise psychrotolerant mesophilic communities. Shifts in the population structure of archaeal (methanogenic) communities were more indicative of process disruption than bacterial communities. Biomolecular techniques were demonstrated as valuable tools for anaerobic wastewater treatment plant monitoring.

Granular biomass from a laboratory-scale anaerobic bioreactor trial was analysed to identify changes in microbial community structure and function in response to temperature and trichloroethylene (TCE). Two bioreactors were operated at 37 degrees C, while two were operated at 15 degrees C. At the time of sampling, one of each temperature pair of bioreactors was exposed to process failure-inducing concentrations of TCE (60 mg L-1) while the other served as a TCE-free control. Bacterial community structure was investigated using denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene clone library analysis. Temperature was identified as an important factor for bacterial community composition, while minor differences were associated with trichloroethylene supplementation. Proteobacteria was the dominant phylum in all bioreactors, while clone library analysis revealed a higher proportion of Bacteroidetes-, Chloroflexi-, and Firmicutes-like clones at 15 degrees C than at 37 degrees C. Comparative metaproteomics in the presence and absence of TCE was carried out by two-dimensional gel electrophoresis (2-DGE), and 28 protein spots were identified, with putative functions related to cellular processes, including methanogenesis, glycolysis, the glyoxylate cycle, and the methyl malonyl pathway. A good agreement between metaproteomic species assignment and phylogenetic information was observed, with 10 of the identified proteins associated with members of the phylum Proteobacteria. This work was supported by Enterprise Ireland, The Irish Environmental Protection Agency, and Science Foundation Ireland. peer-reviewed

The feasibility of low-temperature (7 °C) anaerobic digestion for the treatment of a trichloroethylene (TCE) contaminated wastewater was investigated. Two expanded granular sludge bed (EGSB) bioreactors (R1 and R2) were employed for the mineralisation of a synthetic volatile fatty acid based wastewater at an initial organic loading rate (OLR) of 3 kg COD m(-3) d(-1), and an operating temperature of 15 °C. Successive reductions in OLR to 0.75 kg COD m(-3) d(-1), and operational temperature to 7 °C, resulted in stable bioreactor operation by day 417, with COD removal efficiency and biogas CH(4) content ≥ 74%, for both bioreactors. Subsequently, the influent to R1 was supplemented with increasing concentrations (10, 20, 30 mg l(-1)) of TCE, while R2 acted as a control. At an influent TCE concentration of 30 mg l(-1), although phase average TCE removal rates of 79% were recorded, a sustained decrease in R1 performance was observed, with COD removal of 6%, and % biogas CH(4) of 3% recorded on days 595 and 607, respectively. Specific methanogenic activity (SMA) assays identified a general shift from acetate- to hydrogen-mediated methanogenesis in both R1 and R2 biomass, while toxicity assays confirmed an increased sensitivity of the acetoclastic community in R1 to TCE and dichloroethylene (DCE), which contributed to acetate accumulation. Quantitative Polymerase Chain Reaction (qPCR) analysis of the methanogenic community confirmed the dominance of hydrogenotrophic methanogens in both R1 and R2, representing 71-89% of the total methanogenic population, however acetoclastic Methanosaeta were the dominant organisms, based on 16S rRNA gene clone library analysis of reactor biomass. The greatest change in the bacterial community, as demonstrated by UPGMA analysis of DGGE banding profiles, was observed in R1 biomass between days 417 and 609, although 88% similarity was retained between these sampling points.

The impact of a trichloroethylene (TCE) contaminated wastewater on the microbial community structure of an anaerobic granular biomass at 15 °C compared to 37 °C was investigated. Four expanded granular sludge bed (EGSB) bioreactors (R1–R4) were employed in pairs at 37 and 15 °C. The influents of one of each pair were supplemented with increasing concentrations of TCE (max. 60 mg l−1). At 37 °C, stable operation was maintained with 88% COD removal and >99% TCE removal at maximum influent TCE concentrations. R3 performance decreased at influent TCE concentration of 60 mg l−1, although TCE removal rates of >97% were recorded. Archaeal community analysis via clone library and quantitative polymerase chain reaction (qPCR) analysis, and bacterial community analysis via denaturing gradient gel electrophoresis (DGGE), indicated that temperature resulted in a greater change in community structure than the presence of TCE, and clones related to cold adaptation of biomass were identified at 15 °C.

Four expanded granular sludge bed (EGSB) bioreactors were seeded with a mesophilically-grown granular sludge and operated in duplicate for mesophilic (37 °C; R1 & R2) and low- (15°; R3 & R4) temperature treatment of a synthetic volatile fatty acid (VFA) based wastewater (3 kg COD m(-3) d(-1)) with one of each pair (R1 & R3) supplemented with increasing concentrations of trichloroethylene (TCE; 10, 20, 40, 60 mg l(-1)) and one acting as a control. Bioreactor performance was evaluated by % COD removal efficiency and % biogas methane (CH(4)) content. Quantitative Polymerase Chain Reaction (qPCR) was used to investigate the methanogenic community composition and dynamics in the bioreactors during the trial, while specific methanogenic activity (SMA) and toxicity assays were utilized to investigate the activity and TCE/dichloroethylene (DCE) toxicity thresholds of key trophic groups, respectively. At both 37 °C and 15 °C, TCE levels of 60 mg l(-1) resulted in the decline of % COD removal efficiencies to 29% (Day 235) and 37% (Day 238), respectively, and in % biogas CH(4) to 54% (Day 235) and 5% (Day 238), respectively. Despite the inhibitory effect of TCE on the anaerobic digestion process, the main drivers influencing methanogenic community development, as determined by qPCR and Non-metric multidimensional scaling analysis, were (i) wastewater composition and (ii) operating temperature. At the apical TCE concentration both SMA and qPCR of methanogenic archaea suggested that acetoclastic methanogens were somewhat inhibited by the presence of TCE and/or its degradation derivatives, while competition by dechlorinating organisms may have limited the availability of H(2) for hydrogenotrophic methanogenesis. In addition, there appeared to be an inverse correlation between SMA levels and TCE tolerance, a finding that was supported by the analysis of the inhibitory effect of TCE on two additional biomass sources. The results indicate that low-temperature anaerobic digestion is a feasible approach for the treatment of TCE-containing wastewater.

Two expanded granular sludge bed-anaerobic filter (EGSB-AF) bioreactors, R1 and R2, were operated at 15 degrees C for the treatment of toluene-contaminated volatile fatty acid-based wastewater. The seed inoculum and the R1 reactor were unexposed to toluene, prior to and during the trial, respectively. Both reactors were operated at a hydraulic retention time of 24h at applied organic loading rates of 0.71-1.43kg chemical oxygen demand (COD)m(-3)d(-1). Toluene was supplemented to the R2 influent at concentrations of 5-104 mg toluenel(-1) (solubilised in ethanol). Bioreactor performance was evaluated by COD and toluene removal efficiency, and the methane content of biogas (%). Specific methanogenic activity and toxicity assays were employed to investigate the activity and toluene toxicity thresholds of key trophic groups, respectively, within the seed and reactor biomass samples. COD and toluene removal efficiencies of 70-90% and 55-99%, respectively, were achieved during the 630-d trial. Metabolic assays suggested that a psychrotolerant H(2)/CO(2)-utilizing methanogenic community developed in the toluene-degrading biomass. The results indicate the viability of low-temperature anaerobic digestion for the treatment of wastewater containing toluene.

Anaerobic sludge granules were obtained from laboratory-scale anaerobic bioreactors used to treat pharmaceutical-like (methanol-, acetone- and propanol-contaminated) wastewater under low-temperature conditions (15 °C). The microbial diversity and diversity changes of the sludge samples were ascertained by applying 16S rRNA gene cloning and terminal restriction fragment length polymorphism (TRFLP) analyses, respectively, and using sludge samples from the inoculum, throughout and at the conclusion of the bioreactor trial. Data from genetic fingerprinting correlated well with those from physiological activity assays of the reactor biomass. Specifically, for example, TRFLP profiles indicated the dominance of hydrogenotrophic methanogens within the archaeal community, thus supporting the findings of specific methanogenic activity measurements. TRFLP data supported the hypothesis that the deviation between the replicated reactors, in terms of treatment efficiency, was associated with succession within the microbial communities present, and indicated that community development was linked to both operating temperature and wastewater composition. Fluorescence in situ hybridization (FISH) was also applied, to quantitatively assess the abundance of selected microbial groups, and revealed the underestimation of the abundance Methanosarcina by gene cloning analysis and demonstrated the spatial arrangement of these organisms within the architecture of the low-temperature solvent-degrading anaerobic biofilms.

Low-temperature or psychrophilic (

Low-temperature or psychrophilic (

Whole-cell immobilization of selenate-respiring Sulfurospirillum barnesii in polyacrylamide gels was investigated to allow the treatment of selenate contaminated (790 µg Se × L−1) synthetic wastewater with a high molar excess of nitrate (1,500 times) and sulfate (200 times). Gel-immobilized S. barnesii cells were used to inoculate a mesophilic (30°C) bioreactor fed with lactate as electron donor at an organic loading rate of 5 g chemical oxygen demand (COD) × L−1 day−1. Selenate was reduced efficiently (>97%) in the nitrate and sulfate fed bioreactor, and a minimal effluent concentration of 39 µg Se × L−1 was obtained. Scanning electron microscopy with energy dispersive X-ray (SEM–EDX) analysis revealed spherical bioprecipitates of ≤2 µm diameter mostly on the gel surface, consisting of selenium with a minor contribution of sulfur. To validate the bioaugmentation success under microbial competition, gel cubes with immobilized S. barnesii cells were added to an Upflow Anaerobic Sludge Bed (UASB) reactor, resulting in earlier selenate (24 hydraulic retention times (HRTs)) and sulfate (44 HRTs) removal and higher nitrate/nitrite removal efficiencies compared to a non-bioaugmented control reactor. S. barnesii was efficiently immobilized inside the UASB bioreactors as the selenate-reducing activity was maintained during long-term operation (58 days), and molecular analysis showed that S. barnesii was present in both the sludge bed and the effluent. This demonstrates that gel immobilization of specialized bacterial strains can supersede wash-out and out-competition of newly introduced strains in continuous bioaugmented systems. Eventually, proliferation of a selenium-respiring specialist occurred in the non-bioaugmented control reactor, resulting in simultaneous nitrate and selenate removal during a later phase of operation. Electronic supplementary material The online version of this article (doi:10.1007/s00253-009-1915-x) contains supplementary material, which is available to authorized users.