Effects of elevated temperature on bacterial community structure and function in bioreactors treating a synthetic wastewater

The impact of elevated temperature on bacterial community structure and function during aerobic biological wastewater treatment was investigated. Continuous cultures, fed a complex growth medium containing gelatin and α-lactose as the principal carbon and energy sources, supported mixed bacterial consortia at temperatures ranging from 25–65°C. These temperature- and substrate-acclimated organisms were then used as inocula for batch growth experiments in which the kinetics of microbial growth and substrate utilization, efficiency of substrate removal, and mechanism of substrate removal were compared as functions of temperature. Bacterial community analysis by denaturing gradient gel electrophoresis (DGGE) revealed that distinct bacterial consortia were supported at each temperature. The efficiency of substrate removal declined at elevated temperatures. Maximum specific growth rates and the growth yield increased with temperature from 25–45°C, but then decreased with further elevations in temperature. Thus, maximum specific substrate utilization rates did not vary significantly over the 40°C temperature range (0.64 ± 0.04 mg COD mg⁻¹ dry cell mass h⁻¹). A comparison of the degradation of the protein and carbohydrate portions of the feed medium revealed a lag in α-lactose uptake at 55°C, whereas both components were utilized simultaneously at 25°C. Journal of Industrial Microbiology & Biotechnology (2000) 24, 140–145. Received 09 August 1999/ Accepted in revised form 12 November 1999     

Aerobic biological treatment of synthetic municipal wastewater in membrane-coupled bioreactors

Membrane-coupled bioreactors (MBRs) offer many benefits compared to conventional biological wastewater treatment systems; however, their performance characteristics are poorly understood. Laboratory-scale MBRs were used to study bacterial adaptations in physiology and community structure. MBRs were fed a mixture of starch, gelatin, and polyoxyethylene-sorbitan monooleate to simulate the polysaccharide, protein, and lipid components of municipal wastewater. Physiological adaptations were detected by measuring ectoenzyme activity while structural dynamics were studied by denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments. As cell biomass accumulated in the MBRs, pollutant removal efficiency initially improved and then stabilized with respect to effluent concentrations of chemical oxygen demand, protein, and carbohydrate. Comparison of the MBR effluent to filtered reactor fluid indicated that a portion of the observed pollutant removal was due to filtration by the membrane rather than microbial activity. The rates of ectoenzyme-mediated polysaccharide (alpha-glucosidase) and protein (leucine aminopeptidase) hydrolysis became relatively constant once pollutant removal efficiency stabilized. However, the maximum rate of lipid hydrolysis (heptanoate esterase) concomitantly increased more than 10-fold. Similarly, alpha-glucosidase and leucine aminopeptidase ectoenzyme affinities were relatively constant, while the heptanoate esterase affinity increased more than 30-fold. Community analysis revealed that a substantial community shift occurred within the first 7 days of operation. A Flavobacterium-like bacterial population dominated the community (>50% of total band intensity) and continued to do so for the remainder of the experiment.     

Thermophilic aerobic treatment of potato-processing wastewater

Four batches of potato-processing wastewater were collected every two weeks. After sedimentation for 1 h and subsequent decanting, the supernatant had a 5-day biochemical oxygen demand (BOD) ranging from 620 to 1743 mg/l and total suspended solids ranging from 0.31 to 0.49 mg/ml. Thermophilic aerobic digestion (55°C, 0.6l air/min) decreased the BOD and total suspended solids of the supernatant by 98% and 75%, respectively, in 96 h.B. Malladi and S.C. Ingham are with the Department of Applied Microbiology and Food Science, University of Saskatchewan, Saskatoon, Saskatchewan, S7N 0W0, Canada.     

Analysis of microbial communities developed on the fouling layers of a membrane-coupled anaerobic bioreactor applied to wastewater treatment

The structure of the biofouling layers formed on a pilot-scale membrane-coupled upflow anaerobic sludge blanket bioreactor (UASB) used to treat urban wastewater was analyzed by scanning electron microscopy and electron-dispersive X-ray microanalysis. For comparison, control samples of the membranes were fed either UASB effluent or raw wastewater in a laboratory-scale experiment. Microbial diversity in the fouling materials was analyzed by temperature gradient gel electrophoresis (TGGE) combined with sequence analysis of partial 16S rRNA. Significant differences in structure of the Bacteria communities were observed amongst the different fouling layers analyzed in the UASB membranes, particularly following a chemical cleaning step (NaClO), while the Archaea communities retained more similarity in all samples. The main Bacteria populations identified were evolutively close to Firmicutes (42.3%) and Alphaproteobacteria (30.8%), while Archaea were mostly affiliated to the Methanosarcinales and Methanospirillaceae. Sphingomonadaceae-related bacteria and methanogenic Archaea were persistently found as components of biofouling, regardless of chemical cleaning.     

Thermophilic aerobic wastewater treatment,process performance,biomass characteristics,and effluent quality

Thermophilic aerobic wastewater treatment is reviewed. Thermophilic processes have been studied in laboratory and pilot-scale while full-scale applications are rare. The paper focuses on the microbiology of aerobic thermophiles, performance of the aerobic wastewater treatments, sludge yield, and alternatives to enhance performance of thethermophilic process. Thermophilic processes have been shown to operate under markedly high loading rates (30–180 kg COD m⁻³d⁻¹).Reported sludge production values under thermophilic conditions vary between 0.05 and0.3 kg SS kg COD_removed, which are about the same or lower than generally obtained in mesophilic processes. Compared to analogous mesophilic treatment, thermophilic treatment commonly suffers from poorer effluent quality, measured by lower total COD and filtrated (GF-A) COD removals. However, in the removal of soluble (bacterial membrane filtered) COD both mesophilic and thermophilic treatments have produced similar results. Sludge settle ability in thermophilic processes have been reported to be better or poorer than in analogous mesophilic processes, although cases with better settling properties are rare. Combining thermophilic with mesophilic treatment or ultrafiltration may in some cases markedly improve effluent quality. This revised version was published online in June 2006 with corrections to the Cover Date.     

Removal of carbonaceous and nitrogenous pollutants from a synthetic wastewater using a membrane-coupled bioreactor

Two modified Ludzack-Ettinger (MLE)-type membrane-coupled bioreactors (MBRs) were investigated in this study for the purpose of removing both nitrogenous and carbonaceous pollutants from a synthetic wastewater. During the first MBR experiment, removal efficiencies were high (>90%) for chemical oxygen demand (COD) and ammonia, but total nitrogenous pollutant removal efficiency was poor (~25%). Bacterial community analysis of ammonia oxidizing bacteria (AOB) by a nested PCR-DGGE approach detected two Nitrosomonas-like populations and one Nitrosospira-like population. During the initial portion of the second MBR experiment, COD and ammonia removal efficiencies were similar to the first MBR experiment until the COD of the influent wastewater was increased to provide additional electron donors to support denitrification. Total nitrogen removal efficiencies eventually exceeded 90%, with a hydraulic residence time (HRT) of 24 h and a recirculation ratio of 8. When the HRT of the MBR experiment was decreased to 12 h, however, ammonia removal efficiency was adversely affected. A subsequent increase in the HRT to 18 h helped improve removal efficiencies for both ammonia (>85%) and total nitrogenous compounds (~70%). Our research demonstrates that MBRs can be effectively designed to remove both carbonaceous and nitrogenous pollutants. The ability of the microbial community to switch between anoxic (denitrifying) and oxic (nitrifying) conditions, however, represents a critical process constraint for the application of MLE-type MBR systems, such that little benefit is gained compared to conventional designs.     

Biological treatment of a synthetic space mission wastewater using a membrane-aerated,membrane-coupled bioreactor (M2BR)

This paper describes the membrane-aerated, membrane-coupled bioreactor (M2BR), which was developed for wastewater treatment during long-term space missions because it achieves aeration and biomass separation using components that are compatible with microgravity conditions. In the experiments described herein, the M2BR was used to treat a synthetic wastewater formulated by NASA to simulate the wastewater typically collected during space missions. The M2BR was able to achieve more than 90% removal of both chemical oxygen demand (COD) and total nitrogen when it was fed a modified NASA wastewater that had a 4:1 COD to nitrogen ratio. When the full-strength synthetic wastewater was fed to the M2BR (COD:N=1), however, the nitrogenous pollutant removal efficiency was adversely affected because of either insufficient oxygen transfer to support nitrification (an air-fed M2BR) or insufficient electron donor to support denitrification (an oxygen-fed M2BR). In conclusion, the M2BR provides considerable promise for wastewater treatment during long-term space missions, although additional research is needed to identify the best approach to treat the space mission wastewater, which poses a unique challenge because of its low COD:N ratio.     

Analysis of bacteria communities in an up-flow fixed-bed (UFB) bioreactor for treating sulfide in hydrocarbon wastewater

An up-flow fixed-bed (UFB) bioreactor with patented functional polyurethane foam (FPUF) carriers was used to treat sulfide in hydrocarbon wastewater. Community compositions of autotrophic and heterotrophic bacteria were analyzed by polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE). DGGE results showed that a relatively stable bacterial community composed of heterotrophic and autotrophic bacteria formed in the bioreactor by the end of experiment, which ensured 92–100% sulfide removal efficiencies. Furthermore, autotrophic genera of Thiobacillus and Thiomonas, as well as those of the heterotrophic genus of Acinetobacter survived and exhibited high sulfide oxidation activity under all three operational conditions. Different special genera were also observed under each operational condition, such as the halophilic genus of Nesterenkonia. In addition, a new genus of sulfide oxidation bacteria was found in the bioreactor, which had the ability to synthesize cytoplasm from organic compounds. These genera have wide applications for the treatment of sulfide in hydrocarbon wastewater.     

Thermophilic anaerobic treatment of sulphur rich forest industry wastewater

Thermophilic anaerobic treatment of sulphur-rich paper mill wastewater (0.8-3.1 gCOD/l, 340–850 mgSO₄/l; COD:SO₄ 3.4-5.3) was studied in three laboratory-scale, upflow anaerobic sludge blanket (UASB) reactors and in bioassays. The reactors were inoculated with non-adapted thermophilic granular sludge. In the bioassays, no inhibition of the inoculum was detected and about 62% COD removal (sulphide stripped) was obtained. About 70 to 80% of the removed COD was methanised. In the reactors, up to 60–74% COD removal (effluent sulphide stripped) was obtained at loading rates up to 10–30 kgCOD/m³d and hydraulic retention times down to 6 to 2 hours. The effluent total sulphide was up to 150–250 mg/l. Sulphide inhibition could not be confirmed from the reactor performances. The results from bioassays suggested that both the inoculum and sludge from the UASB reactor used acetate mainly for methane production, while sulphide was produced from hydrogen or its precursors.     

A novel membrane bioreactor for detoxifying industrial wastewater: II. Biodegradation of 3-chloronitrobenzene in an industrially produced wastewater

A novel membrane bioreactor has been used for the treatment of an industrially produced wastewater arising in the manufacture of 3-chloronitrobenzene. This wastewater is not amenable to direct biological treatment without some form pretreatment or dilution, due to the inorganic composition (pH <1, salt concentration 4% w/w) of the wastewater. In the membrane bioreactor, the organic pollutants are first separated from the wastewater by selective membrane permeation, and then biodegraded in the biological growth compartment of the bioreactor. At a wastewater flow rate of 64 mL h(-1) (corresponding to a contact time of approximately 1.7 hours) over 99% of the 3-chloronitrobenzene and over 99% of the nitrobenzene in the wastewater were degraded. Degradation of 3-chloronitrobenzene was accompanied by evolution of chloride ions in a stoichiometric ratio. Both 3-chloronitrobenzene and nitrobenzene degradation were accompanied by the evolution of carbon dioxide; approximately 80% of the carbon entering the system was oxidized to CO(2) carbon. Analysis of mass transport across the membrane revealed that 3-chloronitrobenzene and nitrobenzene are transported more rapidly than phenol. This is explained in terms of a resistances-in-series model, which shows phenol transfer to be rate limited by the membrane diffusion step, whereas the chloronitrobenzene and nitrobenzene transfer are rate limited by the liquid film mass transfer. (c) 1993 Wiley & Sons, Inc.     

Biodegradation of BTEX vapors in a silicone membrane bioreactor system

The biotreatment of complex mixtures of volatile organic compounds (VOCs) such as benzene, toluene, ethylbenzene, and xylene isomers (BTEX) has been investigated by many workers. However, the majority of the work has dealt with the treatment of aqueous or soil phase contamination. The biological treatment of gas and vapor phase sources of VOC wastes has recently received attention with increased usage of biofilters and bioscrubbers. Although these systems are relatively inexpensive, performance problems associated with biomass plugging, gas channeling, and support media acidification have limited their adoption. In this report we describe the development and evaluation of an alternative biotreatment system that allows rapid diffusion of both BTEX and oxygen through a silicone membrane to an active biofilm. The bioreactor system has a rapid liquid recycle, which facilitates nutrient medium mixing over the biofilm and allows for removal of sloughing cell mass. The system removed BTEX at rates up to 30 μg h⁻¹ cm⁻² of membrane area. BTEX removal efficiencies ranged from 75% to 99% depending on the BTEX concentration and vapor flowrate. Consequently, the system can be used for continuous removal and destruction of BTEX and other potential target VOCs in vapor phase streams. Journal of Industrial Microbiology & Biotechnology (2001) 26, 316–325. Received 14 August 2000/ Accepted in revised form 28 February 2001     

A novel membrane bioreactor for detoxifying industrial wastewater: I. Biodegradation of phenol in a synthetically concocted wastewater

A novel process has been used to biodegrade phenol present in an acidic (1 M HCI) and salty (5% w/w NaCl) synthetically bioreactor, in which the phenol present in the wastewater is separated from the inorganic components by means of a silicone rubber membrane. Transfer of the phenol from the wastewater and into a biological growth medium allows biodegradation to proceed under controlled conditions which are unaffected by the hostile inorganic composition of the wastewater. At a wastewater flow rate of 18 mL h(-1) (contact time 6 h), 98.5% of the phenol present in the wastewater at an inlet concentration of 1000 mg ( (-1) ) was degraded; at a contact time of 1.9 h, 65% of the phenol was degraded. Phenol degradation was accompanied by growth of a biofilm on the membrane tubes and by conversion of approximately 80% of the carbon entering the system to CO(2) carbon. Analysis of the transport of phenol across the membrane revealed that the major resistance to mass transfer arose in the diffusion of phenol across the silicone rubber membrane. A mathematical model was used to describe the transfer of phenol across the membrane and the subsequent diffusion and reaction of phenol in the biofilm attached to the membrane tube. This analysis showed that (a) the attached biofilm significantly lowers the mass transfer driving force for phenol across the membrane, and (b) oxygen concentration limits the phenol degradation rate in the biofilm. These conclusions from the model are consistent with the experimental results. (c) 1993 Wiley & Sons, Inc.     

Hydrogenotrophic denitrification of synthetic aquaculture wastewater using membrane bioreactor

A hydrogenotrophic denitrification system was evaluated in removing nitrate from synthetic aquaculture wastewater for recirculation purposes. Two membrane bioreactor (MBR) systems, namely, aeration–denitrification system (ADS) and denitrification–aeration system (DAS) were studied with 50 mg/L of influent concentrations for both organic matter and nitrate nitrogen. The DAS achieved better removal efficiency of 91.4% total nitrogen (T-N) and denitrification rate of 363.7 mg/L.day at a HRT of 3 h compared to ADS. Further, there was no nitrite accumulation in the DAS effluent. The nitrite accumulation in ADS effluent was lesser when CO₂ was used as buffer rather than K₂HPO₄ and KH₂PO₄. Estimation of kinetic parameters of hydrogenotrophic bacteria indicated lesser sludge production compared to heterotrophic denitrification. In the DAS, membrane fouling was nonexistent in the aeration reactor that was used to produce the recirculating effluent. On the contrary, membrane fouling was observed in the denitrification reactor that supplied hydrogen to the mixed liquor. Thus, this study demonstrated DAS capability in maintaining the acceptable water quality appropriate for aquaculture, in which a closed recirculating system is typically used.     

Comparing bacterial community fingerprints from white colonizations in Altamira Cave (Spain)

Monitoring bacterial communities is critical for assessing biodeterioration among other processes. This study presents a strategy and an example of comparative analysis of bacterial communities developing in a cave environment, Altamira Cave which contains unique paleolithic paintings. The analyzed question was whether white colonizations discovered throughout the cave corresponded to similar or different bacterial communities. Molecular fingerprints were obtained by PCR–DGGE from DNA and RNA and statistically compared. Results based on DNA analysis showed that a similar bacterial community was present in white colonizations throughout the cave. Fingerprints based on RNA confirmed the similarity of the major metabolically active components of these communities. The proposed procedure confirmed that white colonizations in Altamira Cave were a consequence of the development of a single complex bacterial community, and the method proves to be highly useful for comparative analysis of microbial communities, including biodeteriorating processes and any other comparative analysis of bacterial communities.     

Performance of a submerged membrane bioreactor for the aerobic treatment of abattoir wastewater

The performance of a submerged membrane bioreactor (SMBR) has been investigated for abattoir wastewater (AW) treatment. The chemical oxygen demand (COD) of permeate has not exceeded 25 mg L⁻¹ providing an average COD removal of 98%. Microbiological analysis showed that the SMBR has allowed a complete removal of fecal coliforms, Listeria and Salmonella. A significant reduction in the excess biomass production was also observed. In fact, the yield of biomass production (Yobs) ranged between 0 and 0.106 g suspended solids/g COD removed.The study of the dynamic of bacterial communities using the single strand conformation polymorphism (SSCP) method showed a significant change in the population structure and revealed a correlation between the sludge production yield and the bacterial communities.     

Comparison of the microbial population dynamics and phylogenetic characterization of a CANOXIS reactor and a UASB reactor degrading trichloroethene

AIMS: To understand the microbial ecology underlying trichloethene (TCE) degradation in a coupled anaerobic/aerobic single stage (CANOXIS) reactor oxygenated with hydrogen peroxide (H2O2) and in an upflow anaerobic sludge bed (UASB) reactor. METHODS AND RESULTS: The molecular study of the microbial population dynamics and a phylogenetic characterization were conducted using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). In both reactors, TCE had a toxic effect on two uncultured bacterial populations whereas oxygen favoured the growth of aerobic species belonging to Rhizobiaceae and Dechloromonas. No methanotrophic bacteria were detected when targeting 16S rRNA gene with universal primers. Alternatively, pmo gene encoding the particulate methane monooxygenase of Methylomonas sp. LW21 could be detected in the coupled reactor when H2O2 was supplied at 0.7 g O2 l day(-1). CONCLUSIONS: Methylomonas sp. LW21 that could be responsible for the aerobic degradation of the TCE by-products is not among the predominant bacterial populations in the coupled reactor. It seems to have been outcompeted by heterotrophic bacteria (Rhizobiaceae and Dechloromonas sp.) for oxygen. SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained show the limitations of the coupled reactor examined in this study. Further investigations should focus on the operating conditions of this reactor in order to favour the growth of the methanotrophs.     

Comparison of microporous and nonporous membrane bioreactor systems for the treatment of BTEX in vapor streams

Increased regulatory constraints on industrial releases of atmospheric volatile organic compounds (VOCs) have resulted in an interest in using biofilters, bioscrubbers and air/liquid membranes for treatment of vapor phase waste streams. In this report, we describe the comparison of the use of two fundamentally different types of membrane module systems that allow the rapid diffusion of vapor phase aromatics and oxygen to an active biofilm for subsequent biodegradation. One system used a commercial membrane module containing microporous polypropylene fibers while the other used a nonporous silicone tubing membrane module for the delivery of substrate (a mixture of benzene, ethylbenzene, toluene, and xylenes [BTEX]) and electron acceptor (O₂). Tests of the systems under similar conditions with BTEX in the vapor feed stream showed significant performance advantages for the silicone membrane system. The average surface-area-based BTEX removal rate for the microporous membrane system over 500 h of operation was 7.88 μg h⁻¹ cm⁻² while the rate for the silicone membrane system was 23.87 μg h⁻¹ cm⁻². The percentages of BTEX removal were also consistently better in the silicone membrane system versus the microporous system. Part of the performance problem associated with the microporous membrane system appeared to be internal water condensation and possible plugging of the pores with biomass over time that could not be resolved with vapor phase backflushing. Journal of Industrial Microbiology & Biotechnology (2002) 28, 245–251 DOI: 10.1038/sj/jim/7000235 Received 17 August 2001/ Accepted in revised form 03 December 2001     

Effects of short solids retention time on microbial community in a membrane bioreactor

Effects of operating lab-scale nitrifying membrane bioreactors (MBR) at short solids retention times (SRT = 3, 5 and 10d) were presented with focus on reactor performance and microbial community composition. The process was capable of achieving over 87% removal of ammonia and 95% removal of chemical oxygen demand (COD), almost regardless of SRT. The denaturing gradient gel electrophoresis (DGGE) analysis shown that bacterial communities evolved in time in a similar way at different SRT. The results of clone library analysis indicated that Betaproteobacteria was the dominant bacterial group in all the reactors but there were significant difference of species for different SRT with higher species diversity at longer SRT. Ammonia and COD removal efficiencies were not correlated with the number of bacterial species or their diversity.     

Effects of creatine supplementation on glucose tolerance and insulin sensitivity in sedentary healthy males undergoing aerobic training

Recent findings have indicated that creatine supplementation may affect glucose metabolism. This study aimed to examine the effects of creatine supplementation, combined with aerobic training, on glucose tolerance in sedentary healthy male. Subjects (n = 22) were randomly divided in two groups and were allocated to receive treatment with either creatine (CT) ( approximately 10 g . day over three months) or placebo (PT) (dextrose). Administration of treatments was double blind. Both groups underwent moderate aerobic training. An oral glucose tolerance test (OGTT) was performed and both fasting plasma insulin and the homeostasis model assessment (HOMA) index were assessed at the start, and after four, eight and twelve weeks. CT demonstrated significant decrease in OGTT area under the curve compared to PT (P = 0.034). There were no differences between groups or over time in fasting insulin or HOMA. The results suggest that creatine supplementation, combined with aerobic training, can improve glucose tolerance but does not affect insulin sensitivity, and may warrant further investigation with diabetic subjects.