Uncertainty in Environmental Micropollutant Modeling.

Water pollution policies have been enacted across the globe to minimize the environmental risks posed by micropollutants (MPs). For regulative institutions to be able to ensure the realization of environmental objectives, they need information on the environmental fate of MPs. Furthermore, there is an urgent need to further improve environmental decision-making, which heavily relies on scientific data. Use of mathematical and computational modeling in environmental permit processes for water construction activities has increased. Uncertainty of input data considers several steps from sampling and analysis to physico-chemical characteristics of MP. Machine learning (ML) methods are an emerging technique in this field. ML techniques might become more crucial for MP modeling as the amount of data is constantly increasing and the emerging new ML approaches and applications are developed. It seems that both modeling strategies, traditional and ML, use quite similar methods to obtain uncertainties. Process based models cannot consider all known and relevant processes, making the comprehensive estimation of uncertainty challenging. Problems in a comprehensive uncertainty analysis within ML approach are even greater. For both approaches generic and common method seems to be more useful in a practice than those emerging from ab initio. The implementation of the modeling results, including uncertainty and the precautionary principle, should be researched more deeply to achieve a reliable estimation of the effect of an action on the chemical and ecological status of an environment without underestimating or overestimating the risk. The prevailing uncertainties need to be identified and acknowledged and if possible, reduced. This paper provides an overview of different aspects that concern the topic of uncertainty in MP modeling.

Widely Used Benzalkonium Chloride Disinfectants Can Promote Antibiotic Resistance.

While the misuse of antibiotics has clearly contributed to the emergence and proliferation of resistant bacterial pathogens, with major health consequences, it remains less clear if the widespread use of disinfectants, such as benzalkonium chlorides (BAC), a different class of biocides than antibiotics, has contributed to this problem. Here, we provide evidence that exposure to BAC coselects for antibiotic-resistant bacteria and describe the underlying genetic mechanisms. After inoculation with river sediment, BAC-fed bioreactors selected for several bacterial taxa, including the opportunistic pathogen Pseudomonas aeruginosa, that were more resistant to several antibiotics than their counterparts in a control (no BAC) bioreactor. A metagenomic analysis of the bioreactor microbial communities, confirmed by gene cloning experiments with the derived isolates, suggested that integrative and conjugative elements encoding a BAC efflux pump together with antibiotic resistance genes were responsible for these results. Furthermore, the exposure of the P. aeruginosa isolates to increasing concentrations of BAC selected for mutations in pmrB (polymyxin resistance) and physiological adaptations that contributed to a higher tolerance to polymyxin B and other antibiotics. The physiological adaptations included the overexpression of mexCD-oprJ multidrug efflux pump genes when BAC was added in the growth medium at subinhibitory concentrations. Collectively, our results demonstrated that disinfectants promote antibiotic resistance via several mechanisms and highlight the need to remediate (degrade) disinfectants in nontarget environments to further restrain the spread of antibiotic-resistant bacteria.IMPORTANCE Benzalkonium chlorides (BAC) are biocides broadly used in disinfectant solutions. Disinfectants are widely used in food processing lines, domestic households, and pharmaceutical products and are typically designed to have a different mode of action than antibiotics to avoid interfering with the use of the latter. Whether exposure to BAC makes bacteria more resistant to antibiotics remains an unresolved issue of obvious practical consequences for public health. Using an integrated approach that combines metagenomics of natural microbial communities with gene cloning experiments with isolates and experimental evolution assays, we show that the widely used benzalkonium chloride disinfectants promote clinically relevant antibiotic resistance. Therefore, more attention should be given to the usage of these disinfectants, and their fate in nontarget environments should be monitored more tightly.

A Rieske-Type Oxygenase of Pseudomonas sp. BIOMIG1 Converts Benzalkonium Chlorides to Benzyldimethyl Amine.

Recently, an array of eight genes involved in the biotransformation of benzalkonium chlorides (BACs)-an active ingredient of many disinfectants-to benzyldimethyl amine (BDMA) was identified in the genome of Pseudomonas sp. BIOMIG1, which is a bacterium present in various environments and mineralizes BACs. In this study, we showed that heterologous expression of an oxygenase gene (oxyBAC) present in this gene array in E. coli resulted in formation of BDMA from BACs at a rate of 14 µM h-1. oxyBAC is phylogenetically classified as a Rieske-type oxygenase (RO) and belongs to a group which catalyzes the cleavage of C-N+ bond between either methyl or alkyl ester and a quaternary nitrogen (N) of natural quaternary ammonium compounds such as stachydrine, carnitine, and trimethylglycine. Insertion of two glycines into the Rieske domain and substitution of tyrosine with leucine in the mononuclear iron center differentiate oxyBAC from other ROs that cleave C-N+, and presumably facilitate the cleavage of saturated alkyl chain from quaternary N via N-dealkylation reaction. In addition, unlike other ROs, oxyBAC did not require a specific reductase to function. Our results demonstrate that oxyBAC represents a new member of RO associated with BAC degradation, and have applications for controlling the fate of BACs in the environment.

Similar Microbial Consortia and Genes Are Involved in the Biodegradation of Benzalkonium Chlorides in Different Environments.

Benzalkonium chlorides (BACs) are emerging pollutants. Identification of microorganisms and the genes involved in the biodegradation of BACs is crucial for better understanding the fate of BACs in the environment and developing treatment strategies. Four microbial communities degrading BACs were developed from sewage (SEW), activated sludge (AS), soil (SOIL) and sea sediment (SEA) samples. According to 16S rRNA pyrosequencing and shotgun metagenome sequencing analyses, the most abundant species represented uncharacterized members of the Pseudomonas and Achromobacter genera. BAC biotransformation rates of the enriched microbial communities were 2.8, 3.2, 17.8, and 24.3 µM hr(-1) for SEA, AS, SOIL, and SEW, respectively, and were positively correlated with the relative abundance of a particular Pseudomonas sp. strain, BIOMIG1. The strain BIOMIG1 mineralizes BACs at a rate up to 2.40 µmol hr(-1) 10(-11) cells. Genomes of four BAC degrading and nondegrading BIOMIG1 phenotypes were sequenced and differentially compared with each other. As a result, a gene cluster encoding for transporters, an integrase and a dioxygenase were involved in BAC biotransformation. Our results suggest that BIOMIG1 plays a key role on the fate of BACs in the environment and genes, other than those reported to date, are involved in BAC biotransformation in various habitats.

Effect of alkyl side chain location and cyclicity on the aerobic biotransformation of naphthenic acids.

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 ß-position or a tertiary carbon at the ß- and/or ß'-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.

Long-term exposure to benzalkonium chloride disinfectants results in change of microbial community structure and increased antimicrobial resistance.

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.

Aerobic biotransformation of n-tetradecylbenzyldimethylammonium chloride by an enriched Pseudomonas spp. community.

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.

Biotransformation of nitrosamines and precursor secondary amines under methanogenic conditions.

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.

Biological removal of benzalkonium chlorides from wastewater by immobilized cells of Pseudomonas sp. BIOMIG1 in an up-flow packed bed reactor.

Quaternary ammonium compounds (QACs) are active ingredients of many disinfectants used against SARS-CoV-2 to control the transmission of the virus through human-contact surfaces. As a result, QAC consumption has increased more than twice during the pandemic. Consequently, the concentration of QACs in wastewater and receiving environments may increase. Due to their antimicrobial activity, high levels of QACs in wastewater may cause malfunctioning of biological treatment systems resulting in inadequate treatment of wastewater. In this study, a biocatalyst was produced by entrapping Pseudomonas sp. BIOMIG1 capable of degrading QACs in calcium alginate. Bioactive 3-mm alginate beads degraded benzalkonium chlorides (BACs), a group of QACs, with a rate of 0.47 µM-BACs/h in shake flasks. A bench-scale continuous up-flow reactor packed with BIOMIG1-beads was operated over one and a half months with either synthetic wastewater or secondary effluent containing 2-20 µM BACs at an empty bed contact time (EBCT) ranging between 0.6 and 4.7 h. Almost complete BAC removal was achieved from synthetic and real wastewater at and above 1.2 h EBCT without aeration and effluent recirculation. The microbial community in beads dominantly composed of BIOMIG1 with trace number of Achromobacter spp. after the operation of the reactor with the real wastewater, suggesting that BIOMIG1 over-competed native wastewater bacteria during the operation. This reactor system offers a low cost and robust treatment of QACs in wastewater. It can be integrated to conventional treatment systems for efficient removal of QACs from the wastewater, especially during the pandemic period.

Validation of an Analytical Workflow for the Analysis of Pesticide and Emerging Organic Contaminant Residues in Paddy Soil and Rice.

Contamination of agricultural soil with organic contaminants is a global problem due to the risks associated with food security and ecological sustainability. Besides the use of agrochemicals, hundreds of emerging contaminants enter arable lands through polluted irrigation water. In this study, an analytical workflow based on QuEChERS extraction coupled with LC-MS/MS quantification was applied to measure 65 emerging contaminants (42 pesticides and 23 multiclass industrial chemicals) in soil and rice for the first time. The method was validated on paddy and yard soil and rice plants. A recovery efficiency ranging between 70 and 120% (RSD <20%) was achieved for more than 70% of the analytes. Then, the validated method was used to quantify target contaminants in 22 soil and 9 rice samples collected mainly from paddy fields close to the Ergene River (Turkey), which is a highly polluted river used for irrigation in the region. Pesticide residues were present in all soil samples up to 2.4 mg/kg. However, their concentrations were below their maximum residual limits in rice. Azoxystrobin, prochloraz, propiconazole, imidacloprid, and epoxiconazole were the most frequently detected pesticides. In addition, industrial pollutants such as benzyldimethyldodecylammonium and tris(2-butoxyethyl) phosphate were detected in paddy soil samples at concentrations between 0.1 and 691 µg/kg. Benzyldimethyldodecylammonium and 5-methyl-1H benzotriazole were also measured in rice at concentrations up to 0.26 and 2.13 µg/kg, respectively.

Identification of core micropollutants of Ergene River and their categorization based on spatiotemporal distribution.

Ergene River is heavily utilized for irrigation of fields to grow the main stocks of rice, wheat, and sunflower of Turkey also exported to Europe; therefore, monitoring the river's water quality is crucial for public health. Although the river quality is routinely monitored, the evaluation of pollution based on micropollutants is limited. In this study, we measured 222 organic micropollutants in 300 samples collected from 75 different locations on the Ergene River between August 2017 and May 2018 using direct injection liquid chromatography-tandem spectrometry with optimized scheduled multiple reaction monitoring. In total, 165 micropollutants were detected at a range of concentrations between 1.90 ng/L and 1824.55 µg/L. Sixty-three chemical substances were recurrent micropollutants that were detected at least one location in all seasons. Among them, 41 chemical substances were identified as the core micropollutants of the Ergene River using data-driven clustering methods. Hexa(methoxymethyl)melamine, benzotriazoles, and benzalkonium chlorides were frequently detected core micropollutants with an industrial origin. Besides, diuron, carbendazim, and cadusafos were common pesticides in the river. Core micropollutants were further categorized based on their type of source and environmental behavior using Kurtosis of concentration and load data obtained for each micropollutant. As a result, the majority of the core micropollutants are recalcitrant chemicals either released from a specific source located upstream of the river or have urban and agricultural sources dispersed on the watershed. In this study, we assessed the current state of pollution in the Ergene River at the micropollutant level with a very high spatial resolution and developed a statistical approach to categorize micropollutants that can be used to monitor the extent of pollution and track pollution sources in the river.

Biotransformation of Acetaminophen by intact cells and crude enzymes of bacteria: A comparative study and modelling.

Acetaminophen (APAP), which is an active ingredient of many analgesic drugs, is one of the contaminants of emerging concern in the environment. Although APAP is biodegradable, it is frequently detected in treatment plant effluents, surface water and soil suggesting that there are factors affecting the fate of APAP in the environment. In this study, four strains of bacteria that can degrade APAP were isolated from soil. Those strains belonged to Rhodococcus, Pseudomonas, Flavobacterium, and Sphingobium genera of Bacteria. A series of kinetic experiments were performed on the isolates in shake-flasks to determine biodegradation rate constant as well as the effect of temperature, APAP concentration and cell density on the biodegradation rates. APAP biodegradation follows the first order reaction kinetics which is coupled with cell growth. The specific APAP biodegradation rate constant (k) for all strains was similar and equal to 0.19 ±â€¯0.01 h-1. The temperature, at which APAP biodegradation rate was maximum, was 35 °C. APAP biodegradation rate was linearly correlated with both the initial APAP concentration and the cell density. Initial step of the APAP biodegradation was hydrolysis of the amide bond which resulted in formation and accumulation of p-aminophenol suggesting that aryl acylamidase enzyme is responsible for the biotransformation. In addition, free and immobilized crude enzymes of the isolates transformed APAP at similar rates, comparable to the intact cells. This study showed that APAP biodegradation is achieved by a diverse group of bacteria having a similar enzyme operating at a constant kinetics which is very slow at environmentally relevant APAP concentrations. Natural removal of APAP in the environment is limited by kinetics, therefore APAP-bearing waste streams should be treated in adsorption enhanced biological systems before discharged into the environment.

Complete Genome Sequence of Pseudomonas sp. Strain BIOMIG1BAC, Which Mineralizes Benzalkonium Chloride Disinfectants.

Pseudomonas sp. strain BIOMIG1BAC is an antibiotic-resistant gammaproteobacterium that can completely mineralize different homologs of benzalkonium chloride disinfectants. Here, we report the annotated complete genome sequence of this microorganism, which includes one circular chromosome with a length of 7,675,262 bp.

Mesophilic and thermophilic anaerobic digestion of municipal sludge and fat, oil, and grease.

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 anaerobic biodegradability of municipal sludge and fat, oil, and grease at mesophilic conditions.

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.

Effect of polyelectrolytes and quaternary ammonium compounds on the anaerobic biological treatment of poultry processing wastewater.

Quaternary ammonium compounds (QACs) and polyelectrolytes are extensively used in poultry processing facilities as sanitizing agents and flocculants, respectively. These chemicals may affect the performance of biological treatment systems resulting in low effluent quality. The impact of these chemicals on the anaerobic treatment of poultry processing wastewater (PPWW) samples, collected before and after a solids separation process, was tested in batch assays using a mixed, mesophilic (35 degrees C) methanogenic culture. The results of this study showed that Vigilquat (VQ), a commercial mixture of four QACs, has a high affinity for the organic solids in the PPWW. Cationic and anionic polyelectrolytes, alone or in combination, did not have any adverse effect on the anaerobic biodegradation of PPWW at concentrations typically used in poultry processing facilities (20 and 5 mg/L, respectively). In spite of the high affinity of VQ for the PPWW solids, VQ at a concentration of 50mg/L and above adversely affected the anaerobic degradation of the PPWW, which resulted in a significantly reduced methane production and accumulation of volatile fatty acids. In the absence of any inhibition, the methane yield varied from 0.76 to 0.98 L methane at STP per g volatile solids added. VQ was not biodegraded under the batch, methanogenic conditions used in this study.

Fate and effect of quaternary ammonium compounds on a mixed methanogenic culture.

The potential inhibitory effect of four quaternary ammonium compounds (QACs) and Vigilquat, a commercial sanitizer which is a mixture of the four QACs, was investigated at concentrations up to 100 mg/L using a mixed, mesophilic (35 degrees C) methanogenic culture. Dextrin and peptone were used as the carbon and energy sources. A batch assay conducted at a range of QAC concentrations showed that QACs were inhibitory to methanogens at and above 25 mg/L. Methanogenesis was more susceptible to QAC inhibition than acidogenesis. Adsorption of QACs on biomass was successfully simulated with the Freundlich isotherm equation. The inhibitory effect of Vigilquat on the mixed methanogenic culture was also investigated in a batch reactor fed with dextrin and peptone. Methanogens were inhibited when the total QAC concentration reached 30 mg/L and volatile fatty acids (VFAs) accumulated. However, methane production recovered in 57 days of incubation, and all VFAs were consumed, suggesting that a prolonged incubation period is necessary for the methanogens to overcome the transient inhibition at a relatively low QAC concentration. None of the QACs tested in this study was biodegraded under methanogenic conditions.

Quaternary ammonium disinfectants: microbial adaptation, degradation and ecology.

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.

Prolonged exposure of mixed aerobic cultures to low temperature and benzalkonium chloride affect the rate and extent of nitrification.

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.

Transition of municipal sludge anaerobic digestion from mesophilic to thermophilic and long-term performance evaluation.

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.