Distribution of tetracycline resistance genes in anaerobic treatment of waste sludge: The role of pH in regulating tetracycline resistant bacteria and horizontal gene transfer. - Related Documents




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796301.0000Distribution of tetracycline resistance genes in anaerobic treatment of waste sludge: The role of pH in regulating tetracycline resistant bacteria and horizontal gene transfer. Although pH value has been widely regarded as an important factor that affects resource recovery of waste sludge, the potential influence of diverse pHs on the distribution of tetracycline resistance genes (TRGs) during sludge anaerobic treatment is largely unknown. Here we reported that in the range of pH 4-10, 0.58-1.18 log unit increase of target TRGs was observed at pH 4, compared with that at pH 7, while 0.70-1.31 log unit further removal were obtained at pH 10. Mechanism study revealed that varied pHs not only altered the community structures of tetracycline resistant bacteria (TRB), but also changed their relative abundances, benefitting the propagation (acidic pHs) or attenuation (alkaline pHs) of TRB. Further investigation indicated that the amount and gene-possessing abilities of key genetic vectors for horizontal TRGs transfer were greatly promoted at acidic pHs but restricted under alkaline conditions.201627485281
796410.9999Fate of sulfonamide resistance genes during sludge anaerobic fermentation: Roles of sludge components and fermentation pHs. This study assessed potential effects of two neglected factors (sludge components and pH values) on the fate of sulfonamide (sul) resistance genes during sludge anaerobic fermentation. It was found that sludge with different contents of protein, carbohydrate and humic acid caused no significant changes in the abundances of sul genes. Nevertheless, sul genes were sensitive to pHs (4-10), and the maximum attenuations (0.8-1.1 log unit) were obtained at pH 10. Mechanism exploration indicated that pHs drove the community evolution of sulfonamide resistant bacteria (SRB), most of which were affiliated to the pH-enriched phyla but not the pH-enriched dominant genera. In addition, the relative abundances of SRB were decreased under both acidic and alkaline conditions. Furthermore, the abundances of intI 1 as well as the sul-carrying abilities of plasmid and extracellular DNA were all reduced at test pHs, indicating that the potential of horizontal gene transfer among bacteria was restricted.201931226672
809320.9998Acidic conditions enhance the removal of sulfonamide antibiotics and antibiotic resistance determinants in swine manure. Manure pH may vary depending on its inherent composition or additive contents. However, the effect of pH on the fate of antibiotics and antibiotic resistance determinants in manure remains unclear. This work demonstrated that pH adjustment promoted the removal of different sulfonamide antibiotics (SAs) within swine manure under incubation conditions, which increased from 26-60.8% to 75.0-86.0% by adjusting the initial pH from neutral (7.4) to acidic (5.4-4.8). Acidification was also demonstrated to inhibit the accumulation of antibiotic resistance genes in manure during incubation. Acidified manure contained both lower absolute and relative abundances of sul1 and sul2 than those at a neutral pH like 7.4. Further investigation indicated that acidification promoted the reduction of sul genes in manure by restricting sulfonamide-resistant bacteria (SRB) proliferation and inhibiting IntI1 accumulation. Furthermore, pH adjustment significantly influenced the composition of the manure bacterial community after incubation, which increased Firmicutes and decreased Proteobacteria. Close relationships were observed between pH-induced enrichment of the Firmicutes bacterial phylum, enhanced SAs degradation, and the fates of antibiotic resistance determinants. Overall, lowering the pH of manure promotes the degradation of SAs, decreases sul genes and SRB, and inhibits horizontal sul gene transfer, which could be a simple yet highly-effective manure management option to reduce antibiotic resistance.202032302890
796630.9998How heavy metal stress promotes dissemination of antibiotic resistance genes in the activated sludge process. Heavy metals have been recently revealed as promoters to antibiotic resistance gene (ARG) dissemination in water environment, but their influence on ARG transfer in the activated sludge process has not been clear. In this study, a set of sequencing batch reactors (SBRs) and micro-scale microfluidic chips were established to quantify the impacts of heavy metals (0.5 mM of Pb, 0.1 mM of As, and 0.005 mM of Hg) on the ARG spreading in the activated sludge consortium. Under heavy metal stress, transfer frequencies were 1.7-3.6 folds increase compared to the control. Gram-negative bacteria increased significantly after heavy metal added, which were more prone to receiving resistant plasmid from donors. Meanwhile, the relative expression of genes related to conjugation changed in activated sludge, especially the expression of outer membrane protein and oxidative stress regulatory genes increased by 2.9-7.4 folds and 7.8-13.1 folds, respectively. Furthermore, using microfluidic chips, the dynamics of ARG transfer was observed at single cell level under heavy metal pressure. Heavy metals firstly promoted conjugation and then vertical gene transfer played an important part for ARG spreading. The results provided in-depth understanding of the influence of heavy metals on ARG behavior in the environment.202235724617
719040.9998Dynamics of microbial community and tetracycline resistance genes in biological nutrient removal process. The occurrence of antibiotics in wastewater has become a serious concern due to the possible development of antibiotic resistant bacteria in wastewater treatment process. In order to understand the dynamics of microbial community and tetracycline resistance genes in biological nutrient removal (BNR) process, three lab-scale sequencing batch reactors (SBRs) were operated under the stress of tetracycline. Results indicated that microbial community structure was altered, and tetracycline efflux pump genes were enhanced over 150-day operation in the presence of trace tetracycline of 20 and 50 μg L(-1), respectively. Furthermore, when the initial tetracycline concentrations were increased to 2 and 5 mg L(-1), substantial enhancement of tetracycline resistance was observed, accompanied with a sharp shift in microbial community structure. In this study, horizontal gene transfer was found to be the main mechanism for the development of tetracycline resistance genes under the long-terms stress of trace tetracycline. About 90.34% of the observed variations in tetracycline resistance genes could be explained by the dynamics of potential hosts of tetracycline resistance genes and class 1 integron. It should be noticed that the functional bacteria (e.g. Nitrospira, Dechloromonas, Rhodobacter and Candidatus_Accumulibacter) responsible for nutrient removal were positively correlated with tetracycline resistance, which might promote the prevalence of tetracycline resistance during biological wastewater treatment. Consequently, this study provided in-depth insights into the occurrence and prevalence of tetracycline resistance genes and their microbial hosts in BNR process.201930849601
719350.9998Plasmid-mediated transfer of antibiotic resistance genes and biofilm formation in a simulated drinking water distribution system under chlorine pressure. The effects of disinfectants and plasmid-based antibiotic resistance genes (ARGs) on the growth of microorganisms and the plasmid-mediated transfer of ARGs in the water and biofilm of the drinking water distribution system under simulated conditions were explored. The heterotrophic plate count of the water in reactors with 0.1 mg/L NaClO and NH(2)Cl was higher than in the control groups. There was no similar phenomenon in biofilm. In the water of reactors containing NaClO, the aphA and bla genes were lower than in the antibiotic resistant bacteria group, while both genes were higher in the water of reactors with NH(2)Cl than in the control group. Chloramine may promote the transfer of ARGs in the water phase. Both genes in the biofilm of the reactors containing chlorine were lower than the control group. Correlation analysis between ARGs and water quality parameters revealed that the copy numbers of the aphA gene were significantly positively correlated with the copy numbers of the bla gene in water and significantly negatively correlated in biofilm (p < 0.05). The results of the sequencing assay showed that bacteria in the biofilm, in the presence of disinfectant, were primarily Gram-negative. 1.0 mg/L chlorine decreased the diversity of the community in the biofilm. The relative abundance of some bacteria that may undergo transfer increased in the biofilm of the reactor containing 0.1 mg/L chlorine.202539617560
694860.9998Fate of antibiotic resistance genes and metal resistance genes during the thermophilic fermentation of solid and liquid swine manures in an ectopic fermentation system. Environmental pollution due to resistance genes from livestock manure has become a serious issue that needs to be resolved. However, little studies focused on the removal of resistance genes in simultaneous processing of livestock feces and urine. This study investigated the fate of antibiotic resistance genes (ARGs), metal resistance genes (MRGs), and class 1 integron-integrase gene (intI1) during thermophilic fermentation of swine manure in an ectopic fermentation system (EFS), which has been regarded as a novel system for efficiently treating both feces and urine. The abundances of MRGs and tetracycline resistance genes were 34.44-97.71% lower in the EFS. The supplementation of heavy metals significantly increased the abundance of intI1, with the enhancement effect of copper being more prominent than that of zinc. The highest abundances of resistance genes and intI1 were observed at high Cu levels (A2), indicating that Cu can increase the spreading of resistance genes through integrons. Network analysis revealed the co-occurrence of ARGs, MRGs, and intI1, and these genes potentially shared the same host bacteria. Redundancy analysis showed that the bacterial community explained most of the variations in ARGs, and environmental factors had influences on ARGs abundances by modulating the bacterial community composition. The decreased Sphingomonas, Comamonas, Acinetobacter, Lactobacillus, Bartonella, Rhizobium, and Bacteroides were mainly responsible for the reduced resistance genes. These results demonstrate that EFS can reduce resistance genes in simultaneous processing of livestock feces and urine.202133592372
719270.9998Sulfamethoxazole and COD increase abundance of sulfonamide resistance genes and change bacterial community structures within sequencing batch reactors. The abundant microbial community in biological treatment processes in wastewater treatment plants (WWTPs) may potentially enhance the horizontal gene transfer of antibiotic resistance genes with the presence of antibiotics. A lab-scale sequencing batch reactor was designed to investigate response of sulfonamide resistance genes (sulI, sulII) and bacterial communities to various concentrations of sulfamethoxazole (SMX) and chemical oxygen demand (COD) of wastewater. The SMX concentrations (0.001 mg/L, 0.1 mg/L and 10 mg/L) decreased with treatment time and higher SMX level was more difficult to remove. The presence of SMX also significantly reduced the removal efficiency of ammonia nitrogen, affecting the normal function of WWTPs. All three concentrations of SMX raised both sulI and sulII genes with higher concentrations exhibiting greater increases. The abundance of sul genes was positive correlated with treatment time and followed the second-order reaction kinetic model. Interestingly, these two genes have rather similar activity. SulI and sulII gene abundance also performed similar response to COD. Simpson index and Shannon-Weiner index did not show changes in the microbial community diversity. However, the 16S rRNA gene cloning and sequencing results showed the bacterial community structures varied during different stages. The results demonstrated that influent antibiotics into WWTPs may facilitate selection of ARGs and affect the wastewater conventional treatment as well as the bacteria community structures.201728211331
809180.9998Changes in antibiotic resistance genes and mobile genetic elements during cattle manure composting after inoculation with Bacillus subtilis. This study explored the effects of Bacillus subtilis at four levels (0, 0.5%, 1%, and 2% w/w compost) on the variations in ARGs, mobile genetic elements (MGEs), and the bacterial community during composting. The composting process had a greater impact on ARGs than Bacillus subtilis. The main ARG detected was sul1. The addition of Bacillus subtilis at 0.5% reduced the relative abundances of ARGs, MGEs, and human pathogenic bacteria (by 2-3 logs) in the mature products. Network and redundancy analyses suggested that intI1, Firmicutes, and pH were mainly responsible for the changes in ARGs, thus controlling these factors might help to inhibit the spread of ARGs.201931442833
808190.9998Potential threat of antibiotics resistance genes in bioleaching of heavy metals from sediment. Bioleaching is considered a promising technology for remediating heavy metals pollution in sediments. During bioleaching, the pressure from the metals bioleached is more likely to cause the spread of antibiotic resistance genes (ARGs). The changes in abundance of ARGs in two typical heavy metal bioleaching treatments using indigenous bacteria or functional bacteria agent were compared in this study. Results showed that both treatments successfully bioleached heavy metals, with a higher removal ratio of Cu with functional bacteria agent. The absolute abundances of most ARGs decreased by one log unit after bioleaching, particularly tetR (p = 0.02) and tetX (p = 0.04), and intI1 decreased from 10(6) to 10(4) copies/g. As for the relative abundance, ARGs in the non-agent treatment increased from 3.90 × 10(-4) to 1.67 × 10(-3) copies/16S rRNA gene copies (p = 0.01), and in the treatment with agent, it reached 6.65 × 10(-2) copies/16S rRNA gene copies, and intI1 relative abundance was maintained at 10(-3) copies/16S rRNA gene copies. The relative abundance of ARGs associated with efflux pump mechanism and ribosomal protection mechanism increased the most. The co-occurrence network indicated that Cu bioleached was the environmental factor determining the distribution of ARGs, Firmicutes might be the potential hosts of ARGs. Compared to bioleaching with indigenous bacteria, the addition of functional bacteria agent engendered a decrease in microbial alpha diversity and an increase in the amount of Cu bioleached, resulting in a higher relative abundance of ARGs. Heavy metal pollution can be effectively removed from sediments using the two bioleaching treatments, however, the risk of ARGs propagation posed by those procedures should be considered, especially the treatment with functional bacteria agents. In the future, an economical and efficient green technology that simultaneously reduces both the absolute abundance and relative abundance of ARGs should be developed.202234979232
7549100.9998Fate of antibiotic resistance genes during sludge anaerobic fermentation: Roles of different sludge pretreatment. Excess sludge, the primary by-product of wastewater treatment plants, is the source and sink of antibiotic resistance genes (ARGs). Sludge pretreatments are an indispensable pathway to improve the resource recovery and harmfulness for anaerobic digestion sludge. However, fewer studies have compared the effects of different pretreatment technologies on the distribution of ARGs during anaerobic sludge digestion. Here, this study established seven anaerobic digesters, and four typical ARGs and one integrase gene of class 1 integron (intI1) regarded as the representative mobile genetic elements (MGEs) were examined during the whole anaerobic digestion process. It was found anaerobic digestion could effectively remove ARGs with about 70.86% removal rate of total ARGs. Among these pretreatments, the reduce efficiency of ARGs was the highest in 50 °C pretreatment, followed by oxidant, and the last was acid-alkaline. The microbial community analysis demonstrated the microbial community structure, including ARGs hosts and antibiotic resistant bacteria, was significantly changed and influenced by high temperature pretreatment. In addition, high temperature and K(2)S(2)O(8) observably decrease the level of ROS production. Macro transcriptome analysis indicated that sludge pretreatment, except for 50 °C pretreatment, up-regulated the genes relevant to lyases and transferase, but down-regulated the genes responsible for peroxidase, antioxidant enzymes and T4SS gene. This study emphasized and compared the different sludge pretreatments on the fate of ARGs in anaerobic sludge, and highlighted concerns regarding the environmental and health risks to our society.202439393457
7961110.9998Effect of tetracycline on ammonia and carbon removal by the facultative bacteria in the anaerobic digester of a sewage treatment plant. This study was conducted to see the effect of tetracycline on nitrogen assimilation and carbon removal in an anaerobic digester of a sewage plant. Samples of sewage were collected from an anaerobic digester sludge. Consortium of nitrogen assimilating bacteria were isolated from the sample and its ability to assimilate ammonia at different concentrations of tetracycline was measured along with carbon removal. The results indicate that while high concentrations of tetracycline of more than 100 mg/L delayed the growth of the bacteria, the resistant bacteria grew after a lag period and the removal of nitrogen and carbon was unaffected even at the highest tetracycline concentration of 250 mg/L tested in this study.201830025323
7596120.9998The impact and fate of clarithromycin in anaerobic digestion of waste activated sludge for biogas production. Clarithromycin retained in waste activated sludge (WAS) inevitably enters the anaerobic digestion system. So far, the complex impacts and fate of clarithromycin in continuous operated WAS anaerobic digestion system are still unclear. In this study, two semi-continuous long-term reactors were set up to investigate the effect of clarithromycin on biogas production and antibiotic resistance genes (ARGs) during WAS anaerobic digestion, and a batch test was carried out to explore the potential metabolic mechanism. Experimental results showed that clarithromycin at lower concentrations (i.e., 0.1 and 1.0 mg/L) did not affect biogas production, whereas the decrease in biogas production was observed when the concentration of clarithromycin was further increased to 10 mg/L. Correspondingly, the relative abundance of functional bacteria in WAS anaerobic digestion (i.e., Anaerolineaceae and Microtrichales) was reduced with long-term clarithromycin exposure. The investigation of ARGs suggested that the effect of methylation belonging to the target site modification played a critical role for the anaerobic microorganisms in the expression of antibiotic resistance, and ermF, played dominated ARGs, presented the most remarkable proliferation. In comparison, the role of efflux pump was weakened with a significant decrease of two detected efflux genes. During WAS anaerobic digestion, clarithromycin could be partially degraded into metabolites with lower antimicrobial activity including oleandomycin and 5-O-desosaminyl-6-O-methylerythronolide and other metabolites without antimicrobial activity.202133545126
7195130.9998Insight into effect of high-level cephalexin on fate and driver mechanism of antibiotics resistance genes in antibiotic wastewater treatment system. In the study, antibiotic resistance genes (ARGs) were examined in wastewater and sludge samples to explore the effect of cephalexin (CFX) on the spreading and removal of ARGs in the Expanded Granular Sludge Bed (EGSB) reactor treating antibiotics wastewater. The result showed that the addition of CFX in the wastewater affected the removal amount of β-lactam ARGs and other types ARGs. Besides, the addition of CFX in the wastewater had no obviously effect on total concentration of targeted ARGs in the sludge, but it was related to the accumulation of some typical ARGs. Based on gene cassette array libraries analysis, the diversity of gene cassettes carried by intI1 gene was increased by the addition of CFX in the wastewater. Furthermore, the co-occurrence patterns between ARGs and bacterial genus were also investigated. The results showed the CFX in the wastewater not only affected the number of potential host bacteria of ARGs, but also changed the types of potential host bacteria of ARGs. The correlation analysis of ARG in influent, effluent and sludge showed that, for blaCTX-M, sul2, qnrS and AmpC genes, their removal amount in EGSB reactor treating antibiotic wastewater system might be enhanced by reducing their concentration in the sludge.202032505047
7194140.9998Response of antibiotic resistance genes in constructed wetlands during treatment of livestock wastewater with different exogenous inducers: Antibiotic and antibiotic-resistant bacteria. This work aimed to study the behavior of antibiotic resistance genes (ARGs) in constructed wetlands with different exogenous inducers additions (oxytetracycline and its resistant bacteria) by high-throughput quantitative polymerase chain reaction. Results indicated that constructed wetlands have the potential to reduce ARGs relative abundances in wastewater, and the total ARGs removal efficiency could exceed 60%. ARGs profile in the effluent differed from that in the influent, and that did not directly reflect the export of dominant ARGs in wetland biofilms. Meanwhile, the highest levels of detected numbers and relative abundances of ARGs were 43 and 3.35 × 10(-1) for control system and 44 and 6.40 × 10(-1) for treatment system, respectively, which meant that ARGs generation in wetlands were inevitable, and antibiotic and antibiotic-resistant bacteria from wastewater could indeed promote ARGs abundance in the system. Compared to the single roles of inducers, their synergistic role had a more significant influence on ARGs relative abundance.202032652450
8013150.9998New insight into fates of sulfonamide and tetracycline resistance genes and resistant bacteria during anaerobic digestion of manure at thermophilic and mesophilic temperatures. This study investigated the variations in antibiotic (sulfonamide and tetracycline) resistance genes (ARGs) and resistant bacteria (ARB) during manure anaerobic digestion (AD) at 35 ℃ and 55 ℃, and discussed the mechanisms of variations in ARGs. The AD lasted for 60 days, five ARGs and intI1 each decreased in abundance after AD at the thermophilic temperature, while only half decreased at the mesophilic temperature. On days 10, 30, and 60, sulfonamide and tetracycline ARB were screened on selective media. During thermophilic AD, ARB numbers reduced by 4-log CFUs per gram dry manure, but only by approximately 1-log CFU at the mesophilic temperature. However, ARB composition analysis showed that at either temperature, no significant reduction in identified ARB species was observed. Furthermore, 72 ARB clones were randomly selected to detect the ARGs they harbored, and the results showed that each ARG was harbored by various hosts, and no definitive link existed between ARGs and bacterial species. In addition, by comparison with the identified host by culture method, the host prediction results based on the correlation analysis between ARGs and the bacterial community was proven to be unreliable. Overall, these findings indicated that relationships between ARB and ARGs were intricate.202031685315
7191160.9998The impact of antimicrobials on the efficiency of methane fermentation of sewage sludge, changes in microbial biodiversity and the spread of antibiotic resistance. The study was designed to simultaneously evaluate the influence of high doses (512-1024 µg/g) the most commonly prescribed antimicrobials on the efficiency of anaerobic digestion of sewage sludge, qualitative and quantitative changes in microbial consortia responsible for the fermentation process, the presence of methanogenic microorganisms, and the fate of antibiotic resistance genes (ARGs). The efficiency of antibiotic degradation during anaerobic treatment was also determined. Metronidazole, amoxicillin and ciprofloxacin exerted the greatest effect on methane fermentation by decreasing its efficiency. Metronidazole, amoxicillin, cefuroxime and sulfamethoxazole were degraded in 100%, whereas ciprofloxacin and nalidixic acid were least susceptible to degradation. The most extensive changes in the structure of digestate microbiota were observed in sewage sludge exposed to metronidazole, where a decrease in the percentage of bacteria of the phylum Bacteroidetes led to an increase in the proportions of bacteria of the phyla Firmicutes and Proteobacteria. The results of the analysis examining changes in the concentration of the functional methanogen gene (mcrA) did not reflect the actual efficiency of methane fermentation. In sewage sludge exposed to antimicrobials, a significant increase was noted in the concentrations of β-lactam, tetracycline and fluoroquinolone ARGs and integrase genes, but selective pressure was not specific to the corresponding ARGs.202133831706
7580170.9997Inactivation of antibiotic resistance genes in antibiotic fermentation residues by ionizing radiation: Exploring the development of recycling economy in antibiotic pharmaceutical factory. Antibiotic fermentation residues are a kind of hazardous waste due to the existence of the residual antibiotics and the potential risk to generate antibiotics resistance genes (ARGs). The appropriate treatment and disposal of antibiotic fermentation residues is imperative. In this study ionizing radiation was applied to treat the antibiotic fermentation residues and the removal efficiencies of antibiotic (erythromycin), ARGs (ermB and ermF) and antibiotic resistant bacteria were investigated. The experimental results showed that erythromycin A content in antibiotic fermentation residues decreased by 86% when the dose was 10 kGy. Moreover, the abundance of ermB and ermF reduced by 89% and 98% at 10 kGy irradiation. Over 99% of total bacteria was removed and antibiotic resistant bacteria (ARB) were less than detection limit after 10 kGy irradiation. Ionizing radiation process is a promising technology for simultaneously removing antibiotic and inactivating ARGs and ARB in antibiotic fermentation residues. Moreover, the irradiation at 10 kGy had no significant influence on the macromolecules organic matters (protein, polysaccharides) of the antibiotic fermentation residues, suggesting that the treated fermentative residues can be used as fertilizer, which could provide the technical support for the development of recycling economy in antibiotic pharmaceutical factory.201930691886
7189180.9997Comparative effects of different antibiotics on antibiotic resistance during swine manure composting. This study explored commonly-used antibiotics (lincomycin, chlorotetracycline, sulfamethoxazole, and ciprofloxacin) and their collective effects on antibiotic resistance during composting. In the first 7 days, ciprofloxacin showed the greatest influence on the physicochemical factors among the studied antibiotics; the removal of antibiotic resistance genes (ARGs) in the multiple-antibiotic treatment was significantly less than single-antibiotic treatments; especially, the largest removal of ribosomal protection genes (tetW and tetO) occurred in single ciprofloxacin treatment. In the end of composting, similar removal ratio (29.71-99.79%) of ARGs was achieved in different treatments (p greater than 0.05); Chloroflexi became the main phylum and it was closely associated with ARGs removal based on the network analysis. Potential host bacteria of ARGs varied with different antibiotics; in particular, the presence of multiple antibiotics increased potential host bacteria of ermA, sul1 and tetO. Above all, collective effects of different antibiotics led to the enrichment of antibiotic resistance in the composting.202032712514
8018190.9997Analysis for microbial denitrification and antibiotic resistance during anaerobic digestion of cattle manure containing antibiotic. This study investigated the effects of tylosin (0, 10, and 100 mg/kg dry weight) on the denitrification genes and microbial community during the anaerobic digestion of cattle manure. N(2) emissions were reduced and N(2)O emissions were increased by 10 mg/kg tylosin. Adding 100 mg/kg tylosin increased the emission of both N(2)O and N(2). The different responses of denitrifying bacteria and genes to tylosin may have been due to the presence of antibiotic resistance genes (ARGs). Network analysis indicated that denitrification genes and ARGs had the same potential host bacteria. intI1 was more important for the horizontal transfer of denitrification genes and ARGs during anaerobic digestion than intI2. The anaerobic digestion of manure containing tylosin may increase nitrogen losses and the associated ecological risk.201931326686