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Polyether ionophores, monensin, and salinomycin are commonly used as antiparasitic drugs in broiler production and may be present in broiler litter (bird excreta plus bedding material). Long-term application of broiler litter to pastures may lead to ionophore contamination of surface waters. Because polyether ionophores break down at low pH, we hypothesized that decreasing litter pH with an acidic material such as aluminum sulfate (alum) would reduce ionophore losses to runoff (i.e., monensin and salinomycin concentrations, loads, or amounts lost). We quantified ionophore loss to runoff in response to (i) addition of alum to broiler litter and (ii) length of time between litter application and the first simulated rainfall event. The factorial experiment consisted of unamended (∼pH 9) vs. alum-amended litters (∼pH 6), each combined with simulated rainfall at 0, 2, or 4 wk after litter application. Runoff from alum-amended broiler litter had 33% lower monensin concentration ( < 0.01), 57% lower monensin load ( < 0.01), 48% lower salinomycin concentration ( < 0.01), and 66% lower salinomycin load ( < 0.01) than runoff from unamended broiler litter when averaged across all events of rainfall. Ionophore losses to runoff were also less when rainfall was delayed for 2 or 4 wk after litter application relative to applying rainfall immediately after litter application. While the weather is difficult to predict, our data suggest that ionophore losses in runoff can be reduced if broiler litter applications are made to maximize dry time after application., (Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.)

Disinfectants play an important role in maintaining acceptable health standards by significantly reducing microbial loads as well as reducing, if not eliminating, pathogens. This review focuses on quaternary ammonium compounds (QACs), a widely used class of organic disinfectants. Specifically, it reviews the occurrence, microbial adaptation, and degradation of QACs, focusing on recent reports on the ecology of QAC-degraders, the pathways and mechanisms of microbial adaptation which lead to resistance to QACs, as well as to antibiotics. With the help of culture-dependent and nonculture-dependent tools, as well as advanced analytical techniques, a better understanding of the fate and effect of QACs and their biotransformation products is emerging. Understanding the underlying mechanisms and conditions that result in QAC resistance and biodegradation will be instrumental in the prudent use of existing QAC formulations and foster the development of safer disinfectants. Development and implementation of (bio)technologies for the elimination of QACs from treated wastewater effluents will lessen adverse impacts to both humans and the environment., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The combined effect of benzalkonium chloride (BAC) and prolonged exposure to low temperature on nitrification was investigated. Ammonia oxidation at 22-24°C by an enriched nitrifying culture was inhibited at increasing BAC concentrations and ceased at 15 mg BAC/L. The non-competitive inhibition coefficient was 1.5±0.9 mg BAC/L. Nitrification tests were conducted without and with BAC at 5mg/L using an aerobic, mixed heterotrophic/nitrifying culture maintained at a temperature range of 24-10°C. Maintaining this culture at 10°C for over one month in the absence of BAC, resulted in slower nitrification kinetics compared to those measured when the culture was first exposed to 10°C. BAC was degraded by the heterotrophic population, but its degradation rate decreased significantly as the culture temperature decreased to 10°C. These results confirm the negative impact of quaternary ammonium compounds on the nitrification process, which is further exacerbated by prolonged, low temperature conditions., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A commercial naphthenic acids (NAs) mixture (TCI Chemicals) and five model NA compounds were ozonated in a semibatch mode. Ozonation of 25 and 35 mg/L NA mixture followed pseudo first-order kinetics (k(obs)=0.11±0.008 min(-1); r(2)=0.989) with a residual NAs concentration of about 5 mg/L. Ozone reacted preferentially with NAs of higher cyclicity and molecular weight and decreased both cyclicity and the acute Microtox® toxicity by 3.3-fold. The ozone reactivity with acyclic and monocyclic model NAs varied and depended on other structural features, such as branching and the presence of tertiary or quaternary carbons. Batch aerobic degradation of unozonated NA mixture using a NA-enriched culture resulted in 83% NA removal and a 6.7-fold decrease in toxicity, whereas a combination of ozonation-biodegradation resulted in 89% NA removal and a 15-fold decrease in toxicity. Thus, ozonation of NA-bearing waste streams coupled with biodegradation are effective treatment processes., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Pulp and paper wastewater (PPW) contains significant concentrations of phytosterols, suspected of inducing endocrine disruption in aquatic species. Aerated stabilization basins (ASBs) are commonly used for the treatment of PPW, but phytosterol removal varies among treatment systems. The objective of this study was to better understand the removal processes and biotransformation of phytosterols within an ASB treatment system fed with untreated PPW. PPW settled solids and supernatant fractions showed that phytosterols are primarily associated with settleable solids, which carry phytosterols to ASB sediment where anoxic/anaerobic conditions prevail. Bioassays with supernatant and settled PPW fractions of the raw wastewater conducted under aerobic and anaerobic conditions, respectively, showed that solids disintegration and hydrolysis results in phytosterol release in ASBs. A simulated ASB, fed with PPW and operated for 2.4 years at three hydraulic retention times (HRTs; 22.2, 11.1 and 5.6 d) with total phytosterol and solids loading rates from 10 to 42 μg/L-d and 44-178 mg/L-d, respectively, was used to determine the steady-state effluent quality and sediment characteristics. Although effluent COD and phytosterol concentrations were relatively low and stable (84-88% total COD removal; 82-94% total phytosterol removal) across the range of HRTs tested, sediment COD and phytosterol concentrations increased with increasing loading rate. On average, 51% of the phytosterols entering the ASB were removed via biotransformation, 40% were retained in the sediment, and the remaining 9% exited with the effluent. This study demonstrates the role of sediment as a source of phytosterol release in ASBs and highlights the importance of HRT and the PPW characteristics for predicting phytosterol fate in ASBs.

The veterinary ionophore antibiotics (IPAs) are extensively used as coccidiostats and growth promoters and are released to the environment via land application of animal waste. Due to their propensity to be transported with runoff, IPAs likely end up in surface waters where they are subject to photodegradation. This study is among the first to investigate the photodegradation of three commonly used IPAs, monensin (MON), salinomycin (SAL) and narasin (NAR), under UV and solar irradiation. Results showed that MON was persistent in a deionized (DI) water matrix when exposed to UV and sunlight, whereas SAL and NAR could undergo direct photolysis with a high quantum yield. Water components including nitrate and dissolved organic matter had a great impact on the photodegradation of IPAs. A pseudosteady state kinetic model was successfully applied to predict IPAs' photodegradation rates in real water matrices. Applying LC/MS/MS, multiple photolytic transformation products of IPAs were observed and their structures were proposed. The direct photolysis of SAL and NAR occurred via cleavage on the ketone moiety and self-sensitized photolysis. With the presence of nitrate, MON was primarily degraded by hydroxyl radicals, whereas SAL showed reactivity toward both hydroxyl and nitrogen-dioxide radicals. Additionally, toxicity tests showed that photodegradation of SAL eliminated its antibiotic properties against Bacillus subtilis.

Veterinary ionophore antibiotics (IPAs) are polyether compounds used extensively in the livestock industry to promote animal growth and prevent coccidia infection. However, the environmental fate and impact of IPAs are not fully understood. In this study, the inhibition and biotransformation potential of the most commonly used IPAs, monensin (MON) and salinomycin (SAL), were investigated under well-defined aerobic, nitrate-reducing, fermentative/sulfate-reducing, and fermentative/methanogenic conditions. Batch assays were conducted with mixed cultures developed from poultry litter (PL), PL-fertilized soil, and municipal anaerobic sludge. Significant transformation of MON and SAL was observed in aerobic, low-buffer capacity culture series as a result of abiotic acid-catalyzed IPAs hydrolysis induced by nitrification. Biotransformation of IPAs was the main transformation process in aerobic, high-buffer capacity culture series. MON persisted under fermentative/sulfate-reducing conditions, whereas SAL was transformed by fermentative bacteria. Both MON and SAL were stable under nitrate-reducing and methanogenic conditions. At IPAs concentrations up to 1 mg/L, MON inhibited only methanogenesis, whereas SAL did not impact any of the biological processes investigated in this study. Multiple, new primary IPA biotransformation products were observed on LC/MS, and their molecular structures were tentatively identified by analyzing LC/MS/MS fragmentation patterns. Overall, MON and SAL exhibited different inhibition and biotransformation patterns at each redox condition tested, which could greatly influence their fate and impact upon their release into the environment as a result of agricultural activities.

Strategies for the transition of municipal sludge anaerobic digestion from mesophilic to thermophilic were assessed and the long-term stability and performance of thermophilic digesters operated at a solids retention time of 30days were evaluated. Transition from 36°C to 53.3°C at a rate of 3°C/day resulted in fluctuation of the daily gas and volatile fatty acids (VFAs) production. Steady-state was reached within 35days from the onset of temperature increase. Transitions from either 36 or 53.3°C to 60°C resulted in relatively stable daily gas production, but VFAs remained at very high levels (in excess of 5000mg COD/L) and methane production was lower than that of the mesophilic reactor. It was concluded that in order to achieve high VS and COD destruction and methane production, the temperature of continuous-flow, suspended growth digesters fed with mixed municipal sludge should be kept below 60°C., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Benzalkonium chlorides (BACs) are disinfectants widely used in a variety of clinical and environmental settings to prevent microbial infections, and they are frequently detected in nontarget environments, such as aquatic and engineered biological systems, even at toxic levels. Therefore, microbial degradation of BACs has important ramifications for alleviating disinfectant toxicity in nontarget environments as well as compromising disinfectant efficacy in target environments. However, how natural microbial communities respond to BAC exposure and what genes underlie BAC biodegradation remain elusive. Our previous metagenomic analysis of a river sediment microbial community revealed that BAC exposure selected for a low-diversity community, dominated by several members of the Pseudomonas genus that quickly degraded BACs. To elucidate the genetic determinants of BAC degradation, we conducted time-series metatranscriptomic analysis of this microbial community during a complete feeding cycle with BACs as the sole carbon and energy source under aerobic conditions. Metatranscriptomic profiles revealed a candidate gene for BAC dealkylation, the first step in BAC biodegradation that results in a product 500 times less toxic. Subsequent biochemical assays and isolate characterization verified that the putative amine oxidase gene product was functionally capable of initiating BAC degradation. Our analysis also revealed cooperative interactions among community members to alleviate BAC toxicity, such as the further degradation of BAC dealkylation by-products by organisms not encoding amine oxidase. Collectively, our results advance the understanding of BAC aerobic biodegradation and provide genetic biomarkers to assess the critical first step of this process in nontarget environments., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

Aerobic biodegradation of naphthenic acids is of importance to the oil industry for the long-term management and environmental impact of process water and wastewater. The effect of structure, particularly the location of the alkyl side chain as well as cyclicity, on the aerobic biotransformation of 10 model naphthenic acids (NAs) was investigated. Using an aerobic, mixed culture, enriched with a commercial NA mixture (NA sodium salt; TCI Chemicals), batch biotransformation assays were conducted with individual model NAs, including eight 8-carbon isomers. It was shown that NAs with a quaternary carbon at the α- or β-position or a tertiary carbon at the β- and/or β'-position are recalcitrant or have limited biodegradability. In addition, branched NAs exhibited lag periods and lower degradation rates than nonbranched or simple cyclic NAs. Two NA isomers used in a closed bottle, aerobic biodegradation assay were mineralized, while 21 and 35% of the parent compound carbon was incorporated into the biomass. The NA biodegradation probability estimated by two widely used models (BIOWIN 2 and 6) and a recently developed model (OCHEM) was compared to the biodegradability of the 10 model NAs tested in this study as well as other related NAs. The biodegradation probability estimated by the OCHEM model agreed best with the experimental data and was best correlated with the measured NA biodegradation rate.

Phytosterols are plant-derived sterols present in pulp and paper wastewater and have been implicated in the endocrine disruption of aquatic species. Bioassays were performed to assess the effect of an additional carbon source and/or solubilizing agent on the aerobic biotransformation of a mixture of three common phytosterols (β-sitosterol, stigmasterol and campesterol). The aerobic biotransformation of the phytosterol mixture by a mixed culture developed from a pulp and paper wastewater treatment system was examined under three separate conditions: with phytosterols as the sole added carbon source, with phytosterols and dextrin as an additional carbon source, and with phytosterols added with ethanol as an additional carbon source and solubilizing agent. Significant phytosterol removal was not observed in assays set up with phytosterol powder, either with or without an additional carbon source. In contrast, all three phytosterols were aerobically degraded when added as a dissolved solution in ethanol. Thus, under the experimental conditions of this study, the bioavailability of phytosterols was limited without the presence of a solubilizing agent. The total phytosterol removal rate was linear for the first six days before re-spiking, with a rate of 0.47 mg/L-d (R(2) = 0.998). After the second spiking, the total phytosterol removal rate was linear for seven days, with a rate of 0.32 mg/L-d (R(2) = 0.968). Following the 7th day, the phytosterol removal rate markedly accelerated, suggesting two different mechanisms are involved in phytosterol biotransformation, more likely related to the production of enzyme(s) involved in phytosterol degradation, induced under different cell growth conditions. β-sitosterol was preferentially degraded, as compared to stigmasterol and campesterol, although all three phytosterols fell below detection limits by the 24th day of incubation., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The biotransformation potential of three phytosterols (campesterol, stigmasterol and β-sitosterol) under denitrifying, sulfate-reducing and fermentative/methanogenic conditions was assessed. Using a group contribution method, the standard Gibbs free energy of phytosterols was calculated and used to perform theoretical energetic calculations. The oxidation of phytosterols under aerobic, nitrate-reducing, sulfate-reducing and methanogenic conditions was determined to be energetically feasible. However, using semi-continuously fed cultures maintained at 20-22 °C over 16 weekly feeding cycles (112 days; retention time, 21 days), phytosterol removal was observed under nitrate-reducing and sulfate-reducing conditions, but not under fermentative/methanogenic conditions. Under sulfate-reducing conditions, stigmast-4-en-3-one was identified as an intermediate of phytosterol biotransformation, a reaction more likely carried out by dehydrogenases/isomerases, previously reported to act on cholesterol under both oxic and anoxic (denitrifying) conditions. Further study of the biotransformation of phytosterols under anoxic/anaerobic conditions is necessary to delineate the factors and conditions leading to enhanced phytosterol biodegradation and the development of effective biological treatment systems for the removal of phytosterols from pulp and paper wastewaters and other phytosterol-bearing waste streams.

Ionophore antibiotics (IPAs) are polyether compounds used in broiler feed to promote growth and control coccidiosis. Most of the ingested IPAs are excreted into broiler litter (BL), a mixture of excreta and bedding material. BL is considered a major source of IPAs released into the environment as BL is commonly used to fertilize agricultural fields. This study investigated IPA biodegradation in BL and soil microcosms, as a process affecting the fate of IPAs in the environment. The study focused on the most widely used IPAs, monensin (MON), salinomycin (SAL), and narasin (NAR). MON was stable in BL microcosms at 24-72% water content (water/wet litter, w/w) and 35-60 °C, whereas SAL and NAR degraded under certain conditions. Factor analysis was conducted to delineate the interaction of water and temperature on SAL and NAR degradation in the BL. A major transformation product of SAL and NAR was identified. Abiotic reaction(s) were primarily responsible for the degradation of MON and SAL in nonfertilized soil microcosms, whereas biodegradation contributed significantly in BL-fertilized soil microcosms. SAL biotransformation in soil microcosms yielded the same product as in the BL microcosms. A new primary biotransformation product of MON was identified in soil microcosms. A field study showed that MON and SAL were stable during BL stacking, whereas MON degraded after BL was applied to grassland. The biotransformation product of MON was also detected in the top soil layer where BL was applied.

The inhibitory effects and biotransformation potential of the fluoroquinolone antibiotic ciprofloxacin (CIP) under anoxic (i.e., nitrate reducing) and anaerobic (i.e., sulfate reducing and methanogenic) conditions were investigated. Fermentation and sulfate reduction was inhibited in 10-80 mg/L CIP-amended sulfate-reducing cultures but recovered with prolonged incubation. Methanogenesis in the mixed culture was significantly inhibited at 80-100 mg CIP/L. No significant decrease of CIP concentration was observed under both sulfate-reducing and methanogenic conditions. However, a low degree of CIP biotransformation was observed in a fed-batch denitrifying culture after a lag time even though the microbial, denitrifying activity was gradually inhibited at 24-40 mg CIP/L. Furthermore, the degradation of CIP was accelerated with a CIP reamendment of the denitrifying culture. Two CIP biotransformation products in the denitrifying culture were detected and their proposed chemical structures suggest that the antibiotic quinolone moiety of CIP was intact., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The inhibitory effect of two widely used quaternary ammonium compounds (QACs)--alkyl benzyl dimethyl (AB) and hexadecyl trimethyl (HD) ammonium chloride--on fermentation, methanogenesis and pentachloroaniline (PCA) dechlorination was assessed using a mixed, methanogenic, PCA-dechlorinating culture amended with AB or HD at a concentration range from 5 to 70 μM. PCA dechlorination was inhibited at 5 μM AB and was completely inhibited at 25 or 5 μM by AB or HD, respectively. However, the PCA dechlorination pathway was the same in both the QACs-free and QACs-amended culture series. Fermentation (acidogenesis) and methanogenesis were inhibited by both AB and HD at and above 25 μM but to a lesser degree than PCA dechlorination. Overall, HD resulted in a more severe inhibition of the mixed culture than AB. Adsorption of both QACs to the mixed culture biomass followed the Freundlich isotherm model. The adsorption affinity of HD for the mixed culture biomass was significantly higher than that of AB, which may be related to the observed higher inhibitory effects of HD compared to AB. Both AB and HD were not degraded in the mixed, dechlorinating culture used in this study., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Ionophore antibiotics (IPAs) are widely used as coccidiostats in poultry and other livestock industries to promote growth and prevent infections. Because most of the ingested IPAs are excreted in poultry litter, which is primarily applied as grassland fertilizer, a significant amount of IPAs can be released into the litter-soil-water environment. A robust analytical method has been developed to quantify IPAs (monensin (MON), salinomycin (SAL) and narasin (NAR)) in complex environmental compartments including surface runoff, soil and poultry litter, with success to minimize matrix interference. The method for water samples involves solid-phase extraction (SPE) followed by liquid-liquid extraction (LLE) post-clean up steps. The method for solid samples involves bi-solvent LLE. IPAs were detected by HPLC-MS, with optimized parameters to achieve the highest sensitivity. Nigericin (NIG), an IPA not used in livestock industry, is successfully applied and validated as a surrogate standard. The method recoveries were at 92-95% and 81-85% in runoff samples from unfertilized and litter-fertilized fields, respectively. For solids, the method recoveries were at 93-99% in soils, and 79-83% in poultry litter samples. SAL was detected at up to 22mg/kg and MON and NAR at up to 4mg/kg in broiler litter from different farms. Up to 183μg/kg of MON was detected in litter-fertilized soils. All three IPAs were detected in the rainfall runoff from litter-fertilized lands at concentrations up to 9μg/L., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Quaternary ammonium compounds (QACs) represent widely used cationic biocides that persist in natural environments. Although microbial degradation, sensitivity and resistance to QACs have been extensively documented, a quantitative understanding of how whole communities adapt to QAC exposure remain elusive. To gain insights into these issues, we exposed a microbial community from a contaminated river sediment to varied levels of benzalkonium chlorides (BACs, a family of QACs) for 3 years. Comparative metagenomic analysis showed that the BAC-fed communities were dramatically decreased in phylogenetic diversity compared with the control (no BAC exposure), resulting presumably from BAC toxicity, and dominated by Pseudomonas species (> 50% of the total). Time-course metagenomics revealed that community adaptation occurred primarily via selective enrichment of BAC-degrading Pseudomonas populations, particularly P. nitroreducens, and secondarily via amino acid substitutions and horizontal transfer of a few selected genes in the Pseudomonas populations, including a gene encoding a PAS/PAC sensor protein and ring-hydroxylating dioxygenase genes. P. nitroreducens isolates were reproducibly recoverable from communities after prolonged periods of no-BAC exposure, suggesting that they are robust BAC-degraders. Our study provides new insights into the mechanisms and tempo of microbial community adaptation to QAC exposure and has implications for treating QACs in biological engineered systems., (© 2013 John Wiley & Sons Ltd and Society for Applied Microbiology.)

The biotransformation potential of a commercial naphthenic acid (NA) mixture (NA sodium salt; TCI Chemicals) under aerobic conditions was investigated using mixed aerobic cultures developed under various levels and duration of NA exposure. A culture enriched using the commercial NA mixture as the sole carbon source degraded NAs in a range of NA concentrations, regardless of culture age and the presence of a co-substrate; however, only 28.5% of the NA-carbon was detected as CO2 while 44% was utilized for biomass growth. A fraction of the NA mixture (15-26%) was persistent under all conditions studied. In contrast, a culture fed with a degradable synthetic wastewater only (NA un-amended culture) and another culture fed with the same wastewater and 50 mg NA/L (NA-amended culture), over time lost their ability to degrade NAs. Analysis of the 16S rRNA gene based clone library revealed that 80% of the NA-enriched culture belonged to the γ-Proteobacteria class and was largely dominated by phylotypes most closely related to known NA and hydrocarbon degraders such as Pseudomonas and Microbulbifer. The results of this study indicate that although significant NA degradation is possible, only a small fraction of the NA mixture is completely mineralized to CO2. Further investigation into the biotransformation products and conditions affecting NA biodegradation under realistic refinery and environmental conditions will help to design effective treatment and bioremediation processes., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

A membrane-less microbial fuel cell (ML-MFC) was developed to investigate the simultaneous carbon removal and denitrification. The removal rates of 0.64 kg COD m(-3) of liquid cathode volume (LCV) d(-1) and 0.186 g NO3(-)-N m(-3) of LCV d(-1) were achieved, which resulted in the maximal COD and nitrate removal rates of 100% and 36.7%, respectively. The ML-MFC also achieved a maximal power output of 0.0712 W m(-3) of LCV and 0.844 A m(-3) of LCV in approximately 24h. The maximal coulombic efficiency of anode (CEAn) and cathode (CECa) was 5.1% and 475%, respectively. The anodic gas phase was consisted of 77.2±4.0% CH4, 3.9±0.5% CO2, and 3.9±1.5% N2, which indicated that the low anode coulombic efficiency was due to anodic methane production. The results of this study demonstrated the potential application of ML-MFC in simultaneous carbon and nitrogen removal and energy (electricity) production., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The effect of benzalkonium chlorides (BACs), a widely used class of quaternary ammonium disinfectants, on microbial community structure and antimicrobial resistance was investigated using three aerobic microbial communities: BACs-unexposed (DP, fed a mixture of dextrin/peptone), BACs-exposed (DPB, fed a mixture of dextrin/peptone and BACs), and BACs-enriched (B, fed only BACs). Long-term exposure to BACs reduced community diversity and resulted in the enrichment of BAC-resistant species, predominantly Pseudomonas species. Exposure of the two microbial communities to BACs significantly decreased their susceptibility to BACs as well as three clinically relevant antibiotics (penicillin G, tetracycline, ciprofloxacin). Increased resistance to BACs and penicillin G of the two BACs-exposed communities is predominantly attributed to degradation or transformation of these compounds, whereas resistance to tetracycline and ciprofloxacin is largely due to the activity of efflux pumps. Quantification of several key multidrug resistance genes showed a much higher number of copies of these genes in the DPB and B microbial communities compared to the DP community. Collectively, our findings indicate that exposure of a microbial community to BACs results in increased antibiotic resistance, which has important implications for both human and environmental health.

The adsorption, inhibition, and biotransformation of the fluoroquinolone antibiotic ciprofloxacin (CIP) under aerobic conditions were investigated in this study. The maximum adsorption capacity and the Langmuir constant were 37.9 mg CIP/g VSS and 37 L/g, respectively. A glucose-fed aerobic culture was inhibited by CIP at 10mg/L or higher and the degree of inhibition increased with increasing CIP concentration. However, the microbial activity recovered to some extent with prolonged incubation under a semi-continuous feeding mode. A low extent of CIP biotransformation was observed in an aerobic, glucose-fed culture derived from poultry litter extract. LC/UV/MS analysis of the biotransformation product showed that only the piperazine ring was oxidized, while the antibiotic quinolone part of CIP was intact., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Ionophore antibiotics (IPAs) are polyether antimicrobials widely used in the livestock industry and may enter the environment via land application of animal waste and agricultural runoff. Information is scarce regarding potential transformation of IPAs under environmental conditions. This study is among the first to identify the propensity of IPAs to undergo acid-catalyzed transformation in mildly acidic aquatic systems and characterize the reactions in depth. The study focused on the most widely used monensin (MON) and salinomycin (SAL), and also included narasin (NAR) in the investigation. All three IPAs are susceptible to acid-catalyzed transformation. MON reacts much more slowly than SAL and NAR and exhibits a different kinetic behavior that is further evaluated by a reversible reaction kinetic model. Extensive product characterization identifies that the spiro-ketal group of IPAs is the reactive site for the acid-catalyzed hydrolytic transformation, yielding predominantly isomeric and other products. Toxicity evaluation of the transformation products shows that the products retain some antimicrobial properties. The occurrence of IPAs and isomeric transformation products is also observed in poultry litter and agricultural runoff samples. Considering the common presence of mildly acidic environments (pH 4-7) in soils and waters, the acid-catalyzed transformation identified in this study likely plays an important role in the environmental fate of IPAs.

The fate and effect of the antimicrobial compounds benzalkonium chlorides (BACs) on the biological nitrogen removal (BNR) processes for a continuous-flow, three-stage laboratory-scale BNR system were modeled. Three kinetic sub-models, corresponding to each reactor, were developed and then combined in a comprehensive ASM1-based model. Kinetic parameters for the three sub-models were evaluated using experimental data obtained from independent batch assays. The biodegradation of BACs was modeled with a mixed-substrate Monod equation. The inhibitory effect of BACs on the utilization of degradable COD and denitrification was modeled as competitive inhibition, whereas non-competitive inhibition was used to model the effect of BACs on nitrification and inhibition coefficients were evaluated. The model simulated well the long-term performance of the BNR system treating a poultry processing wastewater with and without BACs. Enhanced BAC degradation by heterotrophs and increased resistance of nitrifiers to BACs, reflecting acclimation/enrichment over time, is a salient feature of the model., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Naphthenic acids (NAs) are a complex group of alkyl-substituted acyclic, monocyclic and polycyclic carboxylic acids present in crude oil, oil sands process water and tailings ponds, as well as in refinery wastewater. Bioassays were performed to investigate the biotransformation potential and inhibitory effect of a commercial NA mixture to nitrification, denitrification and fermentation/methanogenesis using mixed cultures not previously exposed to NAs. NAs inhibited nitrification in a mixed aerobic heterotrophic/nitrifying culture at concentrations as low as 80 mg NA/L, whereas, an enriched nitrifying culture was only affected at 400 mg NA/L. The lower nitrification inhibition in the latter assay is attributed to the higher population size of nitrosofying and nitrifying bacteria compared to the mixed heterotrophic/nitrifying culture. The NA mixture was not inhibitory to denitrifiers up to 400 mg/L. At higher NA concentrations, cell lysis was pronounced and lysis products were the main source of degradable carbon driving denitrification in culture series prepared without an external carbon source. In the presence of a degradable external carbon source, no difference was observed in nitrate reduction rates or nitrogen gas production at all NA concentrations tested. Methanogenesis was completely inhibited at NA concentrations equal to or higher than 200 mg/L. Methanogenic culture series amended with 80 mg NA/L were transiently inhibited and methane production in culture series prepared with NAs and an external carbon source or NAs only recovered in 136 and 41 days, respectively. Accumulation of volatile fatty acids was observed at inhibitory NA concentrations; however, carbon dioxide production was not affected by NAs, indicating that fermentation and acidogenesis were not affected by NAs. NAs were not degraded under nitrate-reducing or fermentative/methanogenic conditions used in the present study, regardless of the presence or not of another, degradable carbon/energy source., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Naphthenic acids (NAs) are a complex group of alkyl-substituted acyclic, monocyclic and polycyclic carboxylic acids present in oil sands process waters, crude oil, refinery wastewater and petroleum products. Crude oil, desalter brine, influent, activated sludge mixed liquor and effluent refinery samples were received from six United States refineries. The total acid number (TAN) of the six crudes tested ranged from 0.12 to 1.5 mg KOH/g crude oil and correlated to the total NA concentration in the crudes. The total NA concentration in the desalter brine, influent, activated sludge mixed liquor and effluent samples ranged from 4.2 to 40.4, 4.5 to 16.6, 9.6 to 140.3 and 2.8 to 11.6 mg NA/L, respectively. The NAs in all wastewater streams accounted for less than 16% of the total COD, indicating that many other organic compounds are present and that NAs are a minor component in refinery wastewaters. Susceptibility tests showed that none of the activated sludge heterotrophic microcosms was completely inhibited by NAs up to 400 mg/L. Growth inhibition ranging from 10 to 59% was observed in all microcosms at and above 100 mg NA/L. NAs chronically-sorbed to activated sludge mixed liquor biomass and powdered activated carbon (PAC) were recalcitrant and persistent. More than 80% of the total NAs remained in the solid phase at the end of the 10-day desorption period (five successive desorption steps). Throughout a 90-day incubation period, the total NA concentration decreased by 33 and 51% in PAC-free and PAC-containing mixed liquor microcosms, respectively. The lower molecular weight fraction of NAs was preferentially degraded in both mixed liquors. The persistence of the residual, higher molecular weight NAs is likely a combination of molecular recalcitrance and decreased bioavailability when chronically-sorbed to the biomass and/or PAC., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The biotransformation of n-tetradecylbenzyldimethylammonium chloride (C(14)BDMA-Cl), a quaternary ammonium compound (QAC), under aerobic conditions by an enriched microbial community growing on benzalkonium chlorides (BACs) was investigated. Biotransformation of C(14)BDMA-Cl commenced with cleavage of the C(alkyl)-N bond and formation of benzyldimethylamine (BDMA). BDMA was further degraded, but in contrast to a previously reported BAC biotransformation pathway, neither benzylmethylamine (BMA) nor benzylamine (BA) was detected as a BDMA biotransformation product. Kinetic assays further confirmed that BMA and BA were not intermediates of C(14)BDMA-Cl transformation by the enriched community. Thus, BDMA is thought to be transformed to dimethylamine and benzoic acid via debenzylation. The biomass-normalized rate of C(14)BDMA-Cl biotransformation was 0.09 μmol/[mg of volatile suspended solids (VSS)·h]. The Microtox acute toxicity EC(50) value of BDMA was 500 times higher than that of C(14)BDMA-Cl. Thus, the aerobic biotransformation of C(14)BDMA-Cl to BDMA results in substantial toxicity reduction. Phylogenetic analysis of Bacteria diversity indicated that the majority of the sequenced clones (98% of the clone library) belonged to the genus Pseudomonas.

Quaternary ammonium compounds (QACs) are used for sanitation in many poultry processing facilities. This work investigated the fate and effect of a mixture of benzalkonium chlorides (BACs), a class of QACs widely used in commercial antimicrobial formulations, on the biological nitrogen removal (BNR) processes. A laboratory-scale BNR system was operated continuously for 670 days, fed with poultry processing wastewater amended with a mixture of BACs. Initially, the nitrogen removal efficiency deteriorated at a BAC feed concentration of 5 mg/L due to the complete inhibition of nitrification. However, after 27 days of operation, the system recovered and achieved 100% ammonia removal. High nitrogen removal efficiency was achieved even after the feed BAC concentration was stepwise increased up to 120 mg/L. Batch nitrification assays performed before, during, and after BAC exposure, showed that rapid microbial acclimation and BAC biodegradation contributed to the recovery of nitrification achieving efficient and stable long-term BNR system operation., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Ester quaternary ammonium compounds (esterquats), which are mainly used as active ingredients in fabric softeners and personal care products, are beginning to replace traditional quaternary ammonium compounds. As a result of hydrophobicity and increasing use, esterquats reach anaerobic treatment systems. However, little is known about the fate of esterquats under anaerobic conditions. In the present study, the potential inhibitory effect and biotransformation of two alkanoylcholines - acetylcholine chloride (ACh-Cl) and lauroylcholine chloride (LCh-Cl) - which are simple esterquats, under methanogenic conditions were investigated. ACh-Cl up to 300 mg/L was not inhibitory to a mixed methanogenic culture. In contrast, methanogenesis was inhibited by LCh-Cl above 50 mg/L, primarily caused by the accumulation of lauric acid which resulted from the abiotic hydrolysis of LCh. Below inhibitory concentrations, both ACh and LCh were transformed to methane by the mixed methanogenic culture. Mass spectrometric analysis confirmed that both alkanoylcholines were first abiotically hydrolyzed to choline and the corresponding alkanoic acid, which were then biotically transformed to methane, carbon dioxide, and ammonia. Thus, alkanoylcholine-containing waste streams can be bioprocessed to form methane, but hydrolysis products such as long-chain alkanoic acids may adversely impact the anaerobic bioconversion of alkanoylcholines., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The feasibility of using a constructed wetland for treatment of nitrate-contaminated groundwater resulting from the land application of biosolids was investigated for a site in the southeastern United States. Biosolids degradation led to the release of ammonia, which upon oxidation resulted in nitrate concentrations in the upper aquifer in the range of 65-400 mg N/L. A laboratory-scale system was constructed in support of a pilot-scale project to investigate the effect of temperature, hydraulic retention time (HRT) and nitrate and carbon loading on denitrification using soil and groundwater from the biosolids application site. The maximum specific reduction rates (MSRR), measured in batch assays conducted with an open to the atmosphere reactor at four initial nitrate concentrations from 70 to 400 mg N/L, showed that the nitrate reduction rate was not affected by the initial nitrate concentration. The MSRR values at 22 °C for nitrate and nitrite were 1.2 ± 0.2 and 0.7 ± 0.1 mg N/mg VSS(COD)-day, respectively. MSRR values were also measured at 5, 10, 15 and 22 °C and the temperature coefficient for nitrate reduction was estimated at 1.13. Based on the performance of laboratory-scale continuous-flow reactors and model simulations, wetland performance can be maintained at high nitrogen removal efficiency (>90%) with an HRT of 3 days or higher and at temperature values as low as 5 °C, as long as there is sufficient biodegradable carbon available to achieve complete denitrification. The results of this study show that based on the climate in the southeastern United States, a constructed wetland can be used for the treatment of nitrate-contaminated groundwater to low, acceptable nitrate levels., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The biotransformation potential of six nitrosamines and their precursor secondary amines by a mixed methanogenic culture was investigated. Among the six nitrosamines tested, N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), and N-nitrosopyrrolidine (NPYR) were almost completely degraded but only when degradable electron donors were available. On the contrary, N-nitrosodiethylamine (NDEA), N-nitrosodipropylamine (NDPA), and N-nitrosodibutylamine (NDBA) were not degraded. Three precursor secondary amines, corresponding to the three biodegradable nitrosamines, were also completely utilized even with very low levels of available electron donors. The secondary amine precursors of the three, nonbiodegradable nitrosamines were also recalcitrant. A bioassay conducted to elucidate the biotransformation pathway of NDMA in the mixed methanogenic culture using H(2) as the electron donor showed that NDMA was utilized as an electron acceptor and transformed to dimethylamine (DMA), which in turn was degraded to ammonia and methane. The H(2) threshold concentration for NDMA bioreduction ranged between 0.0017 and 0.031 atm. Such a high H(2) threshold concentration suggests that in mixed methanogenic cultures, NDMA reducers are weak competitors to other, H(2)-consuming microbial species, such as homoacetogens and methanogens. Thus, complete removal of nitrosamines in anaerobic digestion systems, where the H(2) partial pressure is typically below 10(-4) atm, is difficult to achieve.

The effect of temperature and benzalkonium chloride (BAC) on nitrate reduction was investigated in batch assays using a mixed nitrate reducing culture. Nitrate was transformed completely, mainly through denitrification, to dinitrogen at 5, 10, 15 and 22 °C. In the absence of BAC, reduction of individual nitrogen oxides had different susceptibility to temperature and transient nitrite accumulation was observed at low temperatures. When the effect of BAC was tested up to 100 mg/L from 5 to 22 °C, denitrification was inhibited at and above 50mg BAC/L with transient nitrite accumulation at all temperatures. The effect of BAC was described by a competitive inhibition model. Nitrite reduction was the denitrification step most susceptible to BAC, especially at low temperatures. BAC was not degraded during the batch incubation and was mostly biomass-adsorbed. Overall, this study shows that low temperatures exacerbate the BAC inhibitory effect, which in turn is controlled by adsorption to biomass., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The inhibitory effect and biodegradation of benzalkonium chloride (BAC), a mixture of alkyl benzyl dimethyl ammonium chlorides with different alkyl chain lengths, was investigated at a concentration range from 5 to 20 mg/L and different biomass concentrations in an activated sludge system. A solution containing glucose and mineral salts was used as the wastewater in all the assays performed. The inhibition of respiratory enzymes was identified as the mode of action of BAC as a result of oxygen uptake rate analysis performed at BAC concentrations ranging between 5 and 70 mg/L. The glucose degradation in the activated sludge at different BAC and biomass concentrations was well-described with Monod kinetics with competitive inhibition. The half-saturation inhibition constant (K(I)) which is equivalent to EC(50) of BAC for the activated sludge tested ranged between 0.12 and 3.60 mg/L. The high K(I) values were recorded at low BAC-to-biomass ratios, i.e. less than 10 mg BAC/g VSS, at which BAC was almost totally adsorbed to biomass and not bioavailable. BAC degradation started as soon as glucose was totally consumed. Although BAC was almost totally adsorbed on the biomass, it was degraded completely. Therefore, BAC degradation was modeled using two-phase biodegradation kinetics developed in this study. This model involves rapid partitioning of BAC to biomass and consecutive degradation in both aqueous and solid phases. The aqueous phase BAC degradation rate was twenty times, on average, higher than the solid phase degradation rate. The specific aqueous (k(I1)) and solid (k(I2)) phase BAC utilization rate constants were 1.25 and 0.31 mg BAC/g VSS h, respectively. The findings of this study would help to understand the reason of extensive distribution of quaternary ammonium compounds in wastewater treatment plant effluents and in natural water systems although QACs are biodegradable, and develop strategies to avoid their release and accumulation in the environment., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The microbial processes involved in two-phase anaerobic digestion were investigated by operating a laboratory-scale acid-phase (AP) reactor and analyzing two full-scale, two-phase anaerobic digesters operated under mesophilic (35 °C) conditions. The digesters received a blend of primary sludge and waste activated sludge (WAS). Methane levels of 20% in the laboratory-scale reactor indicated the presence of methanogenic activity in the AP. A phylogenetic analysis of an archaeal 16S rRNA gene clone library of one of the full-scale AP digesters showed that 82% and 5% of the clones were affiliated with the orders Methanobacteriales and Methanosarcinales, respectively. These results indicate that substantial levels of aceticlastic methanogens (order Methanosarcinales) were not maintained at the low solids retention times and acidic conditions (pH 5.2-5.5) of the AP, and that methanogenesis was carried out by hydrogen-utilizing methanogens of the order Methanobacteriales. Approximately 43, 31, and 9% of the archaeal clones from the methanogenic phase (MP) digester were affiliated with the orders Methanosarcinales, Methanomicrobiales, and Methanobacteriales, respectively. A phylogenetic analysis of a bacterial 16S rRNA gene clone library suggested the presence of acetate-oxidizing bacteria (close relatives of Thermacetogenium phaeum, 'Syntrophaceticus schinkii,' and Clostridium ultunense). The high abundance of hydrogen consuming methanogens and the presence of known acetate-oxidizing bacteria suggest that acetate utilization by acetate oxidizing bacteria in syntrophic interaction with hydrogen-utilizing methanogens was an important pathway in the second-stage of the two-phase digestion, which was operated at high ammonium-N concentrations (1.0 and 1.4 g/L). A modified version of the IWA Anaerobic Digestion Model No. 1 (ADM1) with extensions for syntrophic acetate oxidation and weak-acid inhibition adequately described the dynamic profiles of volatile acid production/degradation and methane generation observed in the laboratory-scale AP reactor. The model was validated with historical data from the full-scale digesters.

The effect of pentachloronitrobenzene (PCNB) on denitrification was assessed with two denitrifying cultures (PCNB-free control and PCNB-acclimated) developed from a contaminated estuarine sediment. PCNB was transformed to pentachloroaniline (PCA) in the PCNB-acclimated culture repeatedly amended with 0.1 muM PCNB, but further dechlorination or degradation of PCA was not observed for almost 1 year. The effect of PCNB on denitrification was also investigated with the PCNB-free control culture. PCNB at an initial concentration of 13 muM was transformed to PCA simultaneously with nitrate reduction but only after the nitrate concentration was at or below 20 mg N/l. PCNB addition at an initial concentration of 13 muM to the control denitrifying culture developed as PCNB-free culture resulted in a transient accumulation of nitric oxide (NO) and nitrous oxide (N(2)O). Similarly to the PCNB-acclimated culture, PCNB transformation to PCA started when the nitrate concentration decreased to about 20 mg N/l. A low degree of nitro group removal resulting in the formation of pentachlorobenzene (PeCB) was also observed in the control culture when amended with 13 muM PCNB. Further transformation or degradation of PCA was not observed in all cultures maintained under active nitrate reducing conditions. Based on the results of this study, the presence of nitrate at low concentrations in anoxic/anaerobic soil and sediments is not expected to negatively affect the biotransformation of PCNB to PCA, but dechlorination or degradation of PCA is not expected under active nitrate reducing conditions.

The sorptive behavior of four quaternary ammonium compounds (QACs) - hexadecyl trimethyl ammonium chloride (C(16)TMA), dodecyl trimethyl ammonium chloride (C(12)TMA), hexadecyl benzyl dimethyl ammonium chloride (C(16)BDMA), and dodecyl benzyl dimethyl ammonium chloride (C(12)BDMA) - to municipal primary, waste activated, mesophilic digested, and thermophilic digested sludges was assessed at 22 degrees C. Batch adsorption of all four separately tested QACs to primary sludge reached equilibrium within 4h. At a nominal, initial QAC concentration of 300mg/L and a sludge volatile solids concentration of 1g/L, the extent of adsorption was 13, 88, 67, and 89% for the C(12)TMA, C(16)TMA, C(12)BDMA, and C(16)BDMA, respectively, and correlated positively to the QAC hydrophobicity and negatively to their critical micelle concentration. Equilibrium partitioning data were described by the Freundlich isotherm model. The adsorption capacity of the four sludges was very similar. In binary QAC mixtures, QACs with relatively high adsorption affinity and at relatively high aqueous concentrations decreased the adsorption of QACs with a low adsorption affinity. At pH 7, about 40% of the sludge-C(12)TMA desorbed, whereas less than 5% of the sludge-C(16)BDMA desorbed in 10 days. The effect of pH was negligible on the desorption extent of C(12)TMA at a pH range 4-10 over 10 days, whereas increasing the solution pH to 10 resulted in more than 50% desorption of C(16)BDMA. Given the fact that approximately 50% of the municipal biosolids are land-applied in the US, the data of this study would help in the assessment of the fate of QACs and their potential effect on human and environmental health., (Copyright (c) 2009 Elsevier Ltd. All rights reserved.)

The distribution of product isomers during the sequential reductive dechlorination of pentachloroaniline (PCA) and pentachlorobenzene (PeCB) was examined based on calculated thermodynamic, chromatographic, and electronic properties and then compared to the product distribution achieved by enrichment cultures. The dechlorination pathway analysis based on free energy considerations matched 78% and 67% of the experimental results for the sequential reductive dechlorination of chlorobenzenes (CBs) and chloroanilines (CAs), respectively. Chromatographic properties of CBs and CAs were able to explain some but not all of the reactions in the observed dechlorination pathways. Correlations between the observed dechlorination pattern and electronic properties of the parent compounds were able to explain most of the formation of the observed products. Experimentally observed sequential reductive dechlorination of CBs and CAs were similar to predicted dechlorination pathways based on the charge differential values calculated for the carbon-chloride bonds. Chlorine atoms were removed from the carbon atom that has the highest charge differential or the second highest charge differential. However, although thermodynamic, electronic as well as chromatographic properties of the CBs and CAs are certainly important factors, they may not be sufficient to completely describe the sequential microbial reductive dechlorination. Enzymatic specificity, as well as other factors (i.e., culture acclimation, environmental factors) should be considered for the interpretation of observed sequential reductive dehalogenation pathways of haloorganic compounds. This work provides the most comprehensive analysis to date of theoretical factors that control the sequential reductive chlorination of two homologous series of single-ring chloroaromatic species., (2009 Wiley Periodicals, Inc.)

The reductive dechlorination of pentachloroaniline (PCA) was investigated in the absence and presence of sulfate in batch assays using a PCA-dechlorinating mixed anaerobic culture with methanol as the external electron donor at neutral pH and 22 degrees C. PCA at an initial concentration of 7.8 microM was sequentially dechlorinated to dichlorinated anilines in the sulfate-free culture and the culture amended with 300 mg sulfate-S/L. At an initial concentration of 890 mg sulfate-S/L, a higher sulfate reduction rate was achieved, but PCA dechlorination was not observed until the sulfate concentration dropped to about 100 mg S/L. The transient inhibition of PCA is attributed to competition between sulfate reducing and dechlorinating species for electron donor, more likely for H(2) resulting from methanol fermentation. A long-term (118 days) PCA dechlorination assay with the sulfate-amended culture, which included five feeding cycles, resulted in accumulation of both sulfide (886 mg S/L) and acetate (1,900 mg COD/L). Under these conditions, the sulfate reducers were inhibited, while the rate and pathway of PCA dechlorination were not affected. The results of this study show that the rate of sulfate reduction rather than the sulfate concentration alone dictates the outcome of the competition between sulfate reducers and either dechlorinators or methanogens. The findings of the present study have significant implications relative to the fate and transport of PCA and its dechlorination products in sulfate-laden subsurface systems.

The denitrification process was incorporated into the IWA Anaerobic Digestion Model No. 1 (ADM1) in order to account for the effect of denitrification on the methanogenic fermentation process. The model was calibrated and optimized using previously published experimental data and kinetic parameter values obtained with a mixed, mesophilic (35 degrees C) methanogenic culture. Model simulations were used to predict the effect of nitrate reduction on the methanogenic fermentation process in batch, semi-continuous, and continuous flow reactors experiencing operational changes and/or system disturbances. The extended model clearly revealed the importance of substrate competition between denitrifiers and non-denitrifiers as well as the impact of N-oxide inhibition on process interactions between fermentation, methanogenesis, and denitrification.

The biocathode of a microbial fuel cell (MFC) offers a promising potential for the reductive treatment of oxidized pollutants. In this study, we demonstrated biological Cr(VI) reduction in the cathode of a MFC and identified putative Cr(VI) reducing microorganisms. The MFC was continuously monitored for Cr(VI) reduction and power generation. Acetate was provided to the anode compartment as substrate and bicarbonate was added to the cathode compartment as the sole external carbon source. The contribution of biomass decay and abiotic processes on Cr(VI) reduction was minimal, confirming that most of the Cr(VI) reduction was assisted by microbial activity in the cathode, which utilizes electrons and protons generated from the oxidation of acetate in the anode compartment. Relatively fast Cr(VI) reduction was observed at initial Cr(VI) concentrations below 80 mg/L. However, at 80 mg Cr(VI)/L, Cr(VI) reduction was extremely slow. A maximum Cr(VI) reduction rate of 0.46 mg Cr(VI)/g VSS.h was achieved, which resulted in a current and power density of 123.4 mA/m(2) and 55.5 mW/m(2), respectively. The reduced chromium was nondetectable in the supernatant of the catholyte which indicated complete removal of chromium as Cr(OH)(3) precipitate. Analysis of the 16S rRNA gene based clone library revealed that the cathode biomass was largely dominated by phylotypes closely related to Trichococcus pasteurii and Pseudomonas aeruginosa, the putative Cr(VI) reducers.

The anaerobic biodegradability of a mix of municipal primary sludge (PS), thickened waste activated sludge (TWAS) and fat, oil, and grease (FOG) was assessed using semi-continuous feed, laboratory-scale anaerobic digesters operated at mesophilic (35 degrees C) and thermophilic (52 degrees C) temperature. Addition of a large FOG fraction (48% of the total VS load) to a PS+TWAS mix, resulted in 2.95 times larger methane yield, 152 vs. 449 mL methane @ STP/g VS added at 35 degrees C and 2.6 times larger methane yield, 197 vs. 512 mL methane @ STP/g VS added at 52 degrees C. The high FOG organic load fraction was not inhibitory to the process. The results of this study demonstrate the benefit of sludge and FOG codigestion.

The fate of six nitrosamines and their secondary amine precursors and total precursors in three municipal wastewater treatment plants' primary sludge (PS), waste-activated sludge (WAS), and anaerobic digester mixed liquor (ADML) was investigated. N-Nitrosodimethylamine was detected at significant concentrations, with mean concentrations at 678 +/- 302, 394 +/- 322, and 271 +/- 100 ng/L in PS, WAS, and ADML samples, respectively. N-Nitrosopyrrolidine was the other nitrosamine detected in sludge samples but at about an order of magnitude lower concentrations. PS samples also contained the highest concentrations of secondary amines(mostly dimethylamine (DMA) and pyrrolidine) followed by WAS and ADML samples, with mean DMA concentrations at 1280 +/- 689, 210 +/- 266, and 6.2 +/- 3.9 microg/L, respectively. Secondary amines in ADML and some WAS samples accounted for only 20-30% of total nitrosamine precursors underlining the significance of as of yet uncharacterized precursors. Overall, anaerobic sludge digestion was a sink for nitrosamines and secondary amines on the basis of the decreasing trends of these compounds from PS to WAS to ADML after taking mass balances into account An anaerobic bioassay conducted with ADML showed complete degradation of secondary amines even without additional carbon sources, while nitrosamine removal required carbon addition and was directly related to the chemical oxygen demand consumption.

The anaerobic biodegradability of municipal primary sludge, thickened waste activated sludge (TWAS), and fat, oil, and grease (FOG) was assessed using semi-continuous-feed, laboratory-scale anaerobic digesters and compared with the ultimate degradability obtained from 120-day batch digestion at 35 degrees C. In run 1, combined primary sludge and TWAS (40/60%, volatile solids [VS] basis) were fed to digesters operated at mesophilic (35 degrees C) and thermophilic (52 degrees C) temperatures at loading rates of 0.99 and 1.46 g-VS/L x d for primary sludge and TWAS, respectively, and a hydraulic retention time (HRT) of 12 days. The volatile solids destruction values were 25.3 and 30.7% (69 and 83% biodegradable volatile solids destruction) at 35 degrees C and 52 degrees C, respectively. The methane (CH4) yields were 159 and 197 mL at the standard temperature and pressure (STP) conditions of 0 degree C and 1 atm/g-VS added or 632 and 642 mL @ STP/g-VS destroyed at 35 degrees C and 52 degrees C, respectively. In run 2, a mix of primary sludge, TWAS, and FOG (21/31/48%, volatile solids basis) was fed to an acid digester operated at a 1-day HRT, at 35 degrees C, and a loading rate of 52.5 g-VS/L x d. The acid-reactor effluent was fed to two parallel methane-phase reactors operated at an HRT of 12 days and maintained at 35 degrees C and 52 degrees C, respectively. After an initial period of 20 days with near-zero gas production in the acid reactor, biogas production increased and stabilized to approximately 2 mL CH4 @ STP/g-VS added, corresponding to a volatile solids destruction of 0.4%. The acid-phase reactor achieved a 43% decrease in nonsaturated fat and a 16, 26, and 20% increase of soluble COD, volatile fatty acids, and ammonia, respectively. The methane-phase volatile solids destruction values in run 2 were 45 and 51% (85 and 97% biodegradable volatile solids destruction) at 35 degrees C and 52 degrees C, respectively. The methane yields for the methane-phase reactors were 473 and 551 mL @ STP/g-VS added, which is approximately 3 times larger compared with run 1, or 1040 and 1083 mL @ STP/g-VS destroyed, at 35 degrees C and 52 degrees C, respectively. The results indicate that, when co-digesting municipal sludge and FOG, a large FOG organic load fraction could have a profound effect on the methane gas yield.

The effect and transformation potential of benzalkonium chlorides (BAC) under nitrate reducing conditions were investigated at concentrations up to 100 mg/L in batch assays using a mixed, mesophilic (35 degrees C) methanogenic culture. Glucose was used as the carbon and energy source and the initial nitrate concentration was 70 mg N/L Dissimilatory nitrate reduction to ammonia (DNRA) and to dinitrogen (DNRN) were observed at BAC concentrations up to 25 mg/L At and above 50 mg BAC/L, DNRA was inhibited and DNRN was incomplete resulting in accumulation of nitrous oxide. Long-term inhibition of methanogenesis and accumulation of volatile fatty acids were observed at and above 50 mg BAC/L Over 99% of the added BAC was recovered from all cultures except the one amended with 100 mg BAC/L where 37% of the initially added BAC was transformed during the 100 day incubation period. Abiotic and biotic assays performed with 100 mg/L of BAC and 5 mM (in the liquid phase) of either nitrate, nitrite, or nitric oxide demonstrated that BAC transformation was abiotic and followed the modified Hofmann degradation pathway, i.e., bimolecular nucleophilic substitution with nitrite. Alkyl dimethyl amines (tertiary amines) were produced at equamolar levels to BAC transformed, but were not further degraded. This is the first report demonstrating the transformation of BAC under nitrate reducing conditions and elucidating the BAC transformation pathway.

The effect of operational conditions and initial dye concentration on the reductive transformation (decolorization) of the textile dye Reactive Blue 4 (RB4) using zero-valent iron (ZVI) filings was evaluated in batch assays. The decolorization rate increased with decreasing pH and increasing temperature, mixing intensity, and addition of salt (100gL(-1) NaCl) and base (3gL(-1) Na2CO3 and 1gL(-1) NaOH), conditions typical of textile reactive dyebaths. ZVI RB4 decolorization kinetics at a single initial dye concentration were evaluated using a pseudo first-order model. Under dyebath conditions and at an initial RB4 concentration of 1000mgL(-1), the pseudo first-order rate constant (kobs) was 0.029+/-0.006h(-1), corresponding to a half-life of 24.2h and a ZVI surface area-normalized rate constant (kSA) of 2.9x10(-4)Lm(-2)h(-1). However, as the initial dye concentration increased, the kobs decreased, suggesting saturation of ZVI surface reactive sites. Non-linear regression of initial decolorization rate values as a function of initial dye concentration, based on a reactive sites saturation model, resulted in a maximum decolorization rate (Vm) of 720+/-88mgL(-1)h(-1) and a half-saturation constant (K) of 1299+/-273mgL(-1). Decolorization of RB4 via a reductive transformation, which was essentially irreversible (2-5% re-oxidation), is believed to be the dominant decolorization mechanism. However, some degree of RB4 irreversible sorption cannot be completely discounted. The results of this study show that ZVI treatment is a promising technology for the decolorization of commercial, anthraquinone-bearing, spent reactive dyebaths.

The effect of nitrate reduction onthe reductive biotransformation of pentachloronitrobenzene (PCNB), an organochlorine fungicide, was assessed with a mixed fermentative/methanogenic culture enriched from a contaminated estuarine sediment. Glucose and methanol served asthe electron and carbon source. PCNB at an initial concentration of 3 microM was transformed to pentachloroaniline (PCA) simultaneously with nitrate reduction in cultures amended with 10 to 200 mg N/L nitrate. PCA sequentially dechlorinated to dichlorinated anilines (mainly 2,5-DCA) in the nitrate-free control culture, and the culture which was amended with 10 mg N/L nitrate. PCA partially dechlorinated to tetrachloroanilines (TeCAs), and methanogenesis was completely inhibited in the cultures amended with 50-200 mg N/L nitrate, whereas fermentation was only inhibited in the cultures amended with 200 mg N/L nitrate. The impact of nitrate reduction on the sequential dechlorination of PCA was attributed to the production of nitric oxide (NO) and nitrous oxide (N2O). Partial nitrate reduction to ammonia was observed in the cultures amended with 50, 100, or 200 mg N/L nitrate and PCNB (3 microM). Therefore, nitrate concentrations at or above 50 mg N/L lead to accumulation of toxic compounds such as highly chlorinated anilines (i.e., PCA, TeCAs) and denitrification intermediates (i.e., NO, N2O). These findings have significant environmental implications in terms of the fate and transformation of PCNB in subsurface environments where nitrate is present.

Thermophilic soil geobacilli isolated from cool temperate geographical zone environments have been shown to be metabolically inactive under aerobic conditions at ambient temperatures (-5 to 25 degrees C). It is now confirmed that a similar situation exists for their anaerobic denitrification activity. It is necessary therefore to determine the mechanisms that sustain the observed significant viable populations in these soils. Population analysis of thermophiles in rainwater and air samples has shown different species compositions which support the view that long distance global transport and deposition in rainwater is a possible source of replenishment of the soil thermophile populations. Survival experiments using a representative Geobacillus isolate have indicated that while cells lose viability rapidly at most temperatures, populations can increase only when the temperature allows growth to take place at a rate which exceeds death rate. Long term (9-month) experiments at 4 degrees C show population increases which can be accounted for by very slow growth rates complemented by negligible death rates. These results are interpreted in the context of current hypotheses on the biogeography patterns of bacteria.

The anaerobic biodegradability of municipal primary and secondary sludge with increasing levels of partially dewatered fat, oil, and grease (FOG) was assessed using a mixed methanogenic culture at 35 "C. Under batch conditions with an acclimated and enriched microbial population, the sludge loading was 3 kg volatile solids/m3 and the highest FOG loading tested was 1.5 kg volatile solids/m3, resulting in a methane yield of 245 mL methane/g sludge volatile solids added at 35 degrees C and 1010 mL methane/g FOG volatile solids added at 35 degrees C. Under semicontinuous feeding conditions, the sludge and sludge plus FOG loading tested were 3 and 3.75 kg volatile solids/m3-d, respectively. Within 23 days of operation, the volatile fatty acid concentrations were reduced below 200 mg chemical oxygen demand/L (187 mg/L as acetic acid). Enhancement of sludge digestion was observed in those reactors where codigestion of sludge and FOG took place, which was attributed to a higher level of microbial activity maintained in these reactors as a result of FOG degradation. The results of this study demonstrate that beneficial use of FOG through codigestion with municipal sludge is feasible.

The effect of the quaternary ammonium compound, didecyl dimethyl ammonium chloride (DDAC), on nitrate reduction was investigated at concentrations up to 100 mg/L in a batch assay using a mixed, mesophilic (35 degrees C) methanogenic culture. Glucose was used as the carbon and energy source and the initial nitrate concentration was 70 mg N/L. Dissimilatory nitrate reduction to ammonia (DNRA) and to dinitrogen (denitrification) were observed at DDAC concentrations up to 25 mg/L. At and above 50 mg DDAC/L, DNRA was inhibited and denitrification was incomplete resulting in accumulation of nitrous oxide. At DDAC concentrations above 10 mg/L, production of nitrous oxide, even transiently, resulted in complete, long-term inhibition of methanogenesis and accumulation of volatile fatty acids. Fermentation was inhibited at and above 75 mg DDAC/L. DDAC suppressed microbial growth and caused cell lysis at a concentration 50 mg/L or higher. Most of the added DDAC was adsorbed on the biomass. Over 96% of the added DDAC was recovered from all cultures at the end of the 100-days incubation period, indicating that DDAC did not degrade in the mixed methanogenic culture under the conditions of this study.

The preferential utilization of different electron donors and their effects on the nitrate reduction and methanogenesis in a mixed, mesophilic (35 degrees C) methanogenic culture were investigated. Batch methanogenic cultures were fed with dextrin/peptone (D/P), propionate, acetate, and H(2)/CO(2) at an initial COD of 500 mg/L and an initial nitrate concentration of 50 mg N/L. Immediate cessation of methane production was observed in all nitrate-amended cultures. Methane production completely recovered in the D/P- and acetate-fed cultures, and partially recovered or did not recover in the propionate- and H(2)/CO(2)-fed, nitrate-amended cultures, respectively. Accumulation of denitrification intermediates was observed in both the propionate- and H(2)/CO(2)-fed cultures, which resulted in inhibition of fermentation and/or methanogenesis. The fastest and the slowest nitrate reduction were observed in the acetate- and propionate-fed cultures, respectively., (Copyright IWA Publishing 2008.)