Laundry wastewater and domestic sewage pilot-scale anaerobic treatment: Microbial community resilience regarding sulfide production.

In this study, the removal of anionic surfactant Linear Alkylbenzene Sulfonate (LAS) from laundry wastewater was evaluated in co-digestion with domestic sewage, using a pilot-scale Expanded Granular Sludge Bed reactor. Surfactant influent concentration was enhanced from 5 ±â€¯3 mg LAS L-1 (stage I) to 19 ±â€¯10 mg LAS L-1 (stage II) and 36 ±â€¯19 mg LAS L-1 (stage III) throughout reactor operation. Sulfide levels higher than 20 mg L-1 influenced LAS removal efficiency, which decreased from 71% to 55% and 32% in stage I, II and III, respectively. Acclimation of microbial population was verified and higher relative abundance of the genera similar to Cytophaga, Bacteroides, Syntrophus and Syntrophobacter in the early stages (adaptation and stage I) was replaced by higher relative abundance of the genera Anaerophaga, Nitrosovibrio, Sulfurovum and Desulfovibrio in the last stages (stage II and III).

Application of horizontal-flow anaerobic immobilized biomass reactor for bioremediation of acid mine drainage.

The production of low-pH effluent with sulfate and metals is one of the biggest environmental concerns in the mining industry. The biological process for sulfate reduction has the potential to become a low-cost solution that enables the recovery of interesting compounds. The present study analyzed such a process in a horizontal-flow anaerobic immobilized biomass (HAIB) reactor, employing ethanol as the carbon and energy source. Results showed that a maximal efficiency in the removal of sulfate and ethanol could only be obtained by reducing the applied sulfate load (225.1 ± 38 g m(-3) d(-1)). This strategy led to over 75% of chemical oxygen demand (COD) and sulfate removal. Among the COD/SO4(2-) studied ratios, 0.67 showed the most promising performance. The effluent's pH has naturally remained between 6.8 and 7.0 and the complete oxidation of the organic matter has been observed. Corrections of the influent pH or effluent recirculation did not show any significant effect on the COD and sulfate removal efficiency. Species closely related to strains of Clostridium sp. and species of Acidaminobacter hydrogenomorfans and Fusibacter paucivorans that can be related to the process of sulfate reduction were found in the HAIB reactors when the initial pH was 5 and the COD/SO4(2-) ratio increased to 1.0.

Evaluation of anionic surfactant removal in anaerobic reactor with Fe(III) supplementation.

The objective of this study was to evaluate the removal of linear alkylbenzene sulfonate (LAS) associated with Fe(III) supplementation using an expanded granular sludge bed (EGSB) reactor. The reactor was inoculated with a granular sludge and fed with synthetic wastewater containing a specific LAS load rate (SLLR) of 1.5 mg gVS-1 d-1 (∼16.4 mgLAS L-1 influent) and supplied with 7276 µMol L-1 of Fe(III). The biomasses from the inoculum and at the end of the EGSB-Fe operation (127 days) were characterized using 16S rRNA Ion Tag sequencing. An increase of 20% in the removal efficiency was observed compared to reactors without Fe(III) supplementation that was reported in the literature, and the LAS removal was approximately 84%. The Fe(III) reduction was dissimilatory (the total iron concentration in the influent and effluent were similar) and reached approximately 64%. The higher Fe(III) reduction and LAS removal were corroborated by the enrichment of genera, such as Shewanella (only EGSB-Fe - 0.5%) and Geobacter (1% - inoculum; 18% - EGSB-Fe). Furthermore, the enrichment of genera that degrade LAS and/or aromatic compounds (3.8% - inoculum; 29.6% - EGSB-Fe of relative abundance) was observed for a total of 20 different genera.

The comparative advantages of ethanol and sucrose as co-substrates in the degradation of an anionic surfactant: microbial community selection.

The efficiency of linear alkylbenzene sulfonate (LAS) removal from laundry wastewater and the related microbial community was investigated in an anaerobic fluidized bed reactor (AFBR). The AFBR was operated in three stages, in addition to the biomass adaptation stage without LAS (stage I). The stages were differentiated by their supplementary co-substrates: stage II had sucrose plus ethanol, stage III had only ethanol, and stage IV had no co-substrate. The replacement of sucrose plus ethanol with ethanol only for the substrate composition favored the efficiency of LAS removal, which remained high after the co-substrate was removed (stage II: 52 %; stage III: 73 %; stage IV: 77 %). A transition in the microbial community from Comamonadaceae to Rhodocyclaceae in conjunction with the co-substrate variation was observed using ion sequencing analysis. The microbial community that developed in response to an ethanol-only co-substrate improved LAS degradation more than the community that developed in response to a mixture of sucrose and ethanol, suggesting that ethanol is a better option for enriching an LAS-degrading microbial community.

Analysis of a microbial community associated with polychlorinated biphenyl degradation in anaerobic batch reactors.

The degradation of polychlorinated biphenyls (PCBs) was investigated under fermentative-methanogenic conditions for up to 60 days in the presence of anaerobic biomass from a full-scale UASB reactor. The low methane yields in the PCBs-spiked batch reactors suggested that the biomass had an inhibitory effect on the methanogenic community. Reactors containing PCBs and co-substrates (ethanol/sodium formate) exhibited substantial PCB reductions from 0.7 to 0.2 mg mL(-1). For the Bacteria domain, the PCBs-spiked reactors were grouped with the PCB-free reactors with a similarity of 55 %, which suggested the selection of a specific population in the presence of PCBs. Three genera of bacteria were found exclusively in the PCB-spiked reactors and were identified using pyrosequencing analysis, Sedimentibacter, Tissierela and Fusibacter. Interestingly, the Sedimentibacter, which was previously correlated with the reductive dechlorination of PCBs, had the highest relative abundance in the RCS-PCB (7.4 %) and RCS-PCB-PF (12.4 %) reactors. Thus, the anaerobic sludge from the UASB reactor contains bacteria from the Firmicutes phylum that are capable of degrading PCBs.

Potentially toxic metal contamination and microbial community analysis in an abandoned Pb and Zn mining waste deposit.

Bioindicators have been widely used to assess the contamination of lead/zinc (Pb/Zn) in smelter areas, mining waste disposal areas and in areas containing slags from Pb ore smelting. In this context, the analysis involving microorganisms has gained prominence as a complementary tool in studies aimed at assessing contaminated sites. Therefore, the objective of this study was to assess the contamination of Pb, Zn and cadmium (Cd) in an area where a PbZn smelter operated, using the integration of geochemical and microbial data. The experimental analysis was conducted to characterize the soil collected at three different sites (samples NS, EW and EP). The analysis conducted was physicochemical, metal concentration, metal speciation and analysis of the microbial community through high-throughput sequencing technique. Through the results it was observed that the high concentrations of metals altered the bacterial community present in the soil. Differences were noted between the microbial communities according to the sampling site, especially in sample EP, collected at the margin of the dirt road, which presented higher metal concentrations and microbial diversity. The main phyla detected in the samples were: Proteobacteria, Bacteroidetes and Acidobacteria. Bacteria tolerant to the presence of potentially toxic metals (PTM), such as Rhodoplanes, Kaistobacter, Sphingomonas and Flavisolibacter were identified in the analyzed samples. The phylogenetic groups identified in the study area are similar to those obtained in other studies in metal contaminated areas. The differences between the bacterial communities in each sample indicate that the concentration of PTM may have influenced the microbial community in the soil. Thus, it is noted the importance of the integration of geochemical and microbial data to evaluate the impact of the improper disposal of high PTM concentrated slags in natural soils.

Metagenomic analysis of the microbiome in three different bioreactor configurations applied to commercial laundry wastewater treatment.

The taxonomic and functional diversity of three different biological reactors (fluidized bed reactor, FBR; up-flow anaerobic sludge blanket reactor, UASB; and expanded granular sludge bed reactor, EGSB) used for commercial laundry wastewater treatment was investigated using metagenome shotgun sequencing. Metagenomes were sequenced on the Illumina Hiseq platform and were analyzed using MG-RAST, STAMP and PAST software. The EGSB and UASB reactors were more closely related based on taxonomic and functional profiles, likely due to similar granular sludge and procedures adopted to ensure anaerobic conditions. The EGSB and UASB reactors showed a predominance of methanogens and genes related to methanogenesis, with a prevalence of the acetoclastic pathway, in addition to the peripheral and central O2-independent pathways for aromatic compound degradation. By contrast, FBR showed a dominance of aerobic microbiota and pathways for O2-dependent aromatic compound degradation. Therefore, although the reactors showed similar surfactant removal levels, the microbial composition, functional diversity and aromatic compound degradation pathways were significantly distinct.

Microbial diversity and the implications of sulfide levels in an anaerobic reactor used to remove an anionic surfactant from laundry wastewater.

The objective of this study was to evaluate the removal of linear alkylbenzene sulfonate (LAS) from commercial laundry wastewater using an expanded granular sludge bed (EGSB) reactor with two specific LAS loading rates (SLLRs), 1.0 and 2.7 mg LAS gVS(-1)d (-1). The biomass was characterized using denaturing gradient gel electrophoresis (DGGE) and 16S Ion Tag sequencing. Higher LAS removal (92.9%) was observed in association with an SLLR of 1.0 mg LAS gVS(-1) d(-1) than with an SLLR of 2.7 mg LAS gVS(-1) d(-1) (58.6%). A relationship between the S(-2) concentration in the effluent and the surfactant removal efficiency was observed. This result is indicative of the inhibition of LAS-removing microbiota at S(-2) concentrations greater than 20 mg SL(-1). By using DGGE, microbial stratification was observed in the reactor in association with granule size, even though the reactor is considered to be a completely mixed regime. The RDP-classifier identified 175 genera, 33 of which were related to LAS degradation.

Formaldehyde degradation in an anaerobic packed-bed bioreactor.

The development of appropriate technologies for the treatment of formaldehyde discharged into the environment is important to minimize its impact. Aerobic systems have been employed, although alternative anaerobic treatments have also been widely studied, mainly due to their low energy consumption and sludge production. However, toxic substances can lead to disturbances in anaerobic reactors. Some research has already been developed on formaldehyde anaerobic biological treatment, but no consensus has yet been reached about its behavior nor has the most efficient system been identified. Aiming at finding supporting evidence for this issue, therefore, this study investigated the degradation and toxicity of formaldehyde in a Horizontal-Flow Anaerobic Immobilized Sludge Reactor. Formaldehyde concentrations of 26.2-1158.6 mg HCHO/L were applied in the reactor, resulting in formaldehyde and chemical oxygen demand removal efficiencies of 99.7% and 92%, respectively. Volatile fatty acids with up to five carbons, found during the degradation of formaldehyde, are believed to indicate that the degradation followed routes unlike those suggested in the literature, which reports the formation of intermediates such as methanol and formic acid. The Monod kinetic model adhered to the experimental data well, with apparent kinetic parameters estimated as r(app)max) = 2.79 x 10(-3) mg HCHO/mg SSVh and K (app)(s) = 242.8 mg HCHO/L.

Influence of multiple substrates on anaerobic protein degradation in a packed-bed bioreactor.

Data on the influence of substrate composition on the anaerobic degradation of bovine serum albumin (BSA) in a bench-scale packed-bed reactor are presented and discussed from the standpoint of substrate consumption kinetics. The experiments were carried out in a horizontal-flow anaerobic immobilized biomass (HAIB) reactor fed with BSA based substrates. BSA was the sole carbon source in the first one, while the others were composed of BSA, carbohydrates and lipids. In all the experiments, the HAIB reactor was operated at the hydraulic detention time of 4 hours. The reactor's performance was evaluated based on physicochemical and chromatographic analyses and also on microscopy techniques. A kinetic model of irreversible first-order series-parallel reactions with two intermediate products was proposed, allowing evaluation of the microbial consortium's affinity with the substrates and the metabolic compounds formed. As the first-order kinetic model adhered quite well to the experimental data, the initial protein degradation rates (k) were estimated. The presence of carbohydrates and lipids led the initial protein degradation rate to be reduced. However, the system fed with protein and carbohydrates showed higher process stability.

Anaerobic degradation of BTEX in a packed-bed reactor.

A bench-scale horizontal-flow anaerobic immobilized biomass (HAIB) reactor was assayed aiming to verify its performance in degrading benzene, toluene, ethylbenzene and xylene (BTEX). A 138 ml HAIB reactor filled with polyurethane foam matrices containing immobilized anaerobic biomass was initially fed with synthetic substrate containing protein, carbohydrates and lipids. Thereafter, BTEX degradation was evaluated in the presence of the co-solvents ethanol and lineal alkylbenzene sulphonate (LAS), in two sequential experiments. The inlet BTEX concentration ranged from 1.3 to 27.0 mg/L of each compound and outlet concentrations were lower than 0.1 mg BTEX/L for both the experiments with ethanol and LAS. An active enriched microbial consortium was observed in the reactor, containing BTEX-degraders, and also acetogenic, acetotrophic and hydrogenotrophic microorganisms. The results from the most probable number (MPN) tests indicated a decrease in the number of methanogenic archae, while the number of anaerobic microorganisms in the biofilm was maintained during the experimental period. Methanogenic archae were found to represent less than 0.5 % of the total anaerobic organisms in the biomass inside the reactor.

Evaluation of thermophilic anaerobic microbial consortia using fluorescence in situ hybridization (FISH).

In this study we investigated the development of anaerobic biofilms in differential reactors and suspension cultures in batch reactors under thermophilic (55 degrees C) conditions. FISH, SEM, chemical and chromatographic analysis were used. The differential reactors reached 99.6%, 92.3% and 6.7% of acetic acid, COD and sulfate removal efficiencies, respectively, after 166 h of incubation time. The batch reactor reached 95.6% and 31.8% of acetic acid and sulfate removal efficiencies after 675 h, respectively. FISH results showed that bacterial cells rather than archaeal cells dominated biofilms. These cells, detected with the Bacteria specific probe (EUB338), accounted for 61.1% (+/-3.6) of the DAPI-stained cells and resembled acetate-oxidizing rods and Desulfotomaculum morphologies. Archaeal cells, which hybridized to the Archaea specific probe (ARC915), were also detected in biofilm but they accounted for 36.7% (+/-2.9) of the DAPI-stained cells. These cells were similar to Methanosaeta-like and hydrogenotrophic methanogen rods. In the suspension culture, archaeal cells (58.0%+/-3.8) morphologically similar to Methanosarcina and hydrogenotrophic methanogen rods were predominant over bacterial cells (41.0%+/-4.5), which resembled acetate-oxidizing rods and Desulfotomaculum morphologies. The percentage of sulfate-reducing bacteria cells (SRB) ranged from 12.2% (+/-2.5) to 21.7% (+/-2.8) in the biofilms and from 13.3% (+/-3.6) to 21.7% (+/-4.3) of the DAPI stained cells in suspension culture.

Effect of biomass adaptation to the degradation of anionic surfactants in laundry wastewater using EGSB reactors.

Two expanded granular sludge bed reactors were operated. RAB (adapted biomass) was operated in two stages: Stage I, with standard LAS (13.2 mg L(-1)); and Stage II, in which the standard LAS was replaced by diluted laundry wastewater according to the LAS concentration (11.2 mg L(-1)). RNAB (not adapted biomass) had a single stage, using direct wastewater (11.5 mg L(-1)). Thus, the strategy of biomass adaptation did not lead to an increase of surfactant removal in wastewater (RAB-Stage II: 77%; RNAB-Stage I: 78%). By means of denaturing gradient gel electrophoresis, an 80% similarity was verified in the phases with laundry wastewater (sludge bed) despite the different reactor starting strategies. By pyrosequencing, many reads were related to genera of degraders of aromatic compounds and sulfate reducers (Syntrophorhabdus and Desulfobulbus). The insignificant difference in LAS removal between the two strategies was most likely due to the great microbial richness of the inoculum.

The effect of biomass immobilization support material and bed porosity on hydrogen production in an upflow anaerobic packed-bed bioreactor.

The aim of this study was to investigate the effect of the support material used for biomass attachment and bed porosity on the potential generation of hydrogen gas in an anaerobic bioreactor treating low-strength wastewater. For this purpose, an upflow anaerobic packed-bed (UAPB) reactor fed with sucrose-based synthetic wastewater was used. Three reactors with various support materials (expanded clay, vegetal coal, and low-density polyethylene) were operated for hydraulic retention time (HRT) of 0.5 and 2 h. Based on the results obtained, three further reactors were operated with low-density polyethylene as a material support using various bed porosities (91, 75, and 50 %) for an HRT of 0.5 h. The UAPB reactor was found to be a feasible technology for hydrogen production, reaching a maximum substrate-based hydrogen yield of 7 mol H2 mol(-1) sucrose for an HRT of 0.5 h. The type of support material used did not affect hydrogen production or the microbial population inside the reactor. Increasing the bed porosity to 91 % provided a continuous and cyclic production of hydrogen, whereas the lower bed porosities resulted in a reduced time of hydrogen production due to biomass accumulation, which resulted in a decreasing working volume.

Microbial characterization and removal of anionic surfactant in an expanded granular sludge bed reactor.

This study evaluated linear alkylbenzene sulfonate removal in an expanded granular sludge bed reactor with hydraulic retention times of 26 h and 32 h. Sludge bed and separator phase biomass were phylogenetically characterized (sequencing 16S rRNA) and quantified (most probable number) to determine the total anaerobic bacteria and methanogenic Archaea. The reactor was fed with a mineral medium supplemented with 14 mg l(-1)LAS, ethanol and methanol. The stage I-32 h consisted of biomass adaptation (without LAS influent) until reactor stability was achieved (COD removal >97%). In stage II-32 h, LAS removal was 74% due to factors such as dilution, degradation and adsorption. Higher HRT values increased the LAS removal (stage III: 26 h - 48% and stage IV: 32 h - 64%), probably due to increased contact time between the biomass and LAS. The clone libraries were different between samples from the sludge bed (Synergitetes and Proteobacteria) and the separator phase (Firmicutes and Proteobacteria) biomass.

Treatment of linear alkylbenzene sulfonate in a horizontal anaerobic immobilized biomass reactor.

Linear alkylbenzene sulfonate (LAS) is an anionic surfactant widely used to manufacture detergents and found in domestic and industrial wastewater. LAS removal was evaluated in a horizontal anaerobic immobilized biomass reactor. The system was filled with polyurethane foam and inoculated with sludge that was withdrawn from an up flow anaerobic sludge blanket reactor that is used to treat swine wastewater. The reactor was fed with easily degradable substrates and a solution of commercial LAS for 313 days. The hydraulic retention time applied was 12h. The system was initially operated without detergent and resulted to 94% reduction of demand. The mass balance in the system indicated that the LAS removal efficiency was 45% after 18 0days. From the 109 th day to the 254 th day, a removal efficiency of 32% was observed. The removal of LAS was approximately 40% when 1500 mg of LAS were applied in the absence of co-substrates suggesting that the LAS molecules were used selectively. Microscopic analyses of the biofilm revealed diverse microbial morphologies and denaturing gradient gel electrophoresis profiling showed variations in the total bacteria and sulfate-reducing bacteria populations. 16S rRNA sequencing and phylogenetic analyses demonstrated that members of the order Clostridiales were the major components of the bacterial community in the last step of the reactor operation.

Evaluation of the microbial diversity in a horizontal-flow anaerobic immobilized biomass reactor treating linear alkylbenzene sulfonate.

The purpose of this work was to assess the degradation of linear alkylbenzene sulfonate (LAS) in a horizontal-flow anaerobic immobilized biomass (HAIB) reactor. The reactor was filled with polyurethane foam where the sludge from a sanitary sewage treatment was immobilized. The hydraulic detention time (HDT) used in the experiments was of 12 h. The reactor was fed with synthetic substrate (410 mg l(-1) of meat extract, 115 mg l(-1) of starch, 80 mg l(-1) of saccharose, 320 mg l(-1) of sodium bicarbonate and 5 ml l(-1) of salt solution) in the following stages of operation: SI-synthetic substrate, SII-synthetic substrate with 7 mg l(-1) of LAS, SIII-synthetic substrate with 14 mg l(-1) of LAS and SIV-synthetic substrate containing yeast extract (substituting meat extract) and 14 mg l(-1) of LAS, without starch. At the end of the experiment (313 days) a degradation of approximately 35% of LAS was achieved. The higher the concentration of LAS, the greater the amount of foam for its adsorption. This is necessary because the isotherm of LAS adsorption in the foam is linear for the studied concentrations (2 to 50 mg l(-1)). Microscopic analyses of the biofilm revealed diverse microbial morphologies, while Denaturing Gradient Gel Eletrophoresis (DGGE) profiling showed variations in the population of total bacteria and sulphate-reducing bacteria (SRB). The 16S rRNA gene sequencing and phylogenetic analyses revealed that the members of the order Clostridiales were the major components of the bacterial community in the last reactor operation step.

Influence of the carbon source on the anaerobic biomass adhesion on polyurethane foam matrices.

This work focuses on the influence of the source of organic matter on the process of biomass adhesion on polyurethane foam matrices in fixed-bed anaerobic immobilized-sludge reactors. Five experiments were performed in differential 'gradientless' reactors fed with meat extract (protein), glucose, starch, lipids and complex substrate. The polyurethane foam colonization process was monitored temporally in each experiment to identify the amount of biomass buildup, extracellular polymer production and the morphological characteristics of the cells adhering to the support. Different immobilization patterns were observed for the different substrates used. The morphological variety was found to be dependent on the substrate constituents. Polymer excretion was apparently crucial in the colonization process of the polyurethane matrices and was likely related to cell fixation on the support. The production of extracellular polymeric substances speeded up the initial fixation of microorganisms on the polyurethane surface.

Ethanol and toluene removal in a horizontal-flow anaerobic immobilized biomass reactor in the presence of sulfate.

In this study it is reported the operation of a horizontal-flow anaerobic immobilized biomass (HAIB) reactor under sulfate-reducing condition which was also exposed to different amounts of ethanol and toluene. The system was inoculated with sludge taken from up-flow anaerobic sludge blanket (UASB) reactors treating refuses from a poultry slaughterhouse. The HAIB reactor comprised of an immobilized biomass on polyurethane foam and ferrous and sodium sulfate solutions were used (91 and 550 mg/L, respectively), to promote a sulfate-reducing environment. Toluene was added at an initial concentration of 2.0 mg/L followed by an increased range of different amendments (5, 7, and 9 mg/L). Ethanol was added at an initial concentration of 170 mg/L followed by an increased range of 960 mg/L. The reactor was operated at 30(+/-2) degrees C with hydraulic detention time of 12 h. Organic matter removal efficiency was close to 90% with a maximum toluene degradation rate of 0.06 mg(toluene)/mg(vss)/d. Sulfate reduction was close to 99.9% for all-nutritional amendments. Biofilm microscopic characterization revealed a diversity of microbial morphologies and DGGE-profiling showed a variation of bacterial and sulfate reducing bacteria (SRB) populations, which were significantly associated with toluene amendments. Diversity of archaea remained unaltered during the different phases of this experiment. Thus, this study demonstrates that compact units of HAIB reactors, under sulfate reducing conditions, are a potential alternative for in situ aromatics bioremediation.