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803300.9973Fate of pirlimycin and antibiotic resistance genes in dairy manure slurries in response to temperature and pH adjustment. Quantifying the fate of antibiotics and antibiotic resistance genes (ARGs) in response to physicochemical factors during storage of manure slurries will aid in efforts to reduce the spread of resistance when manure is land-applied. The objectives of this study were to determine the effects of temperature (10, 35, and 55 °C) and initial pH (5, 7, 9, and 12) on the removal of pirlimycin and prevalence of ARGs during storage of dairy manure slurries. We collected and homogenized feces and urine from five lactating dairy cows treated with pirlimycin and prepared slurries by mixing manure and sterile water. Aliquots (200 mL) of slurry were transferred and incubated in 400 mL glass beakers under different temperatures (10, 35, and 55 °C) or initial pH (5, 7, 9, and 12). Pirlimycin concentration and abundances of 16S rRNA, mefA, tet(W), and cfxA as indicators of total bacteria and ARGs corresponding to macrolide, tetracycline, and β-lactam resistance, respectively, were analyzed during manure incubation. The thermophilic environment (55 °C) increased the deconjugation and removal of pirlimycin, while the acidic shock at pH 5 increased deconjugation but inhibited removal of pirlimycin, suggesting that the chemical stability of pirlimycin could be affected by temperature and pH. The thermophilic environment decreased mefA relative abundance on day 7 and 28 (P = 0.02 and 0.04), which indicates that the bacteria that encoded mefA gene were not thermotolerant. Although mefA relative abundance was greater at the pH 9 shock than the rest of pH treatments on day 7 (P = 0.04), no significant pH effect was observed on day 28. The tet(W) abundance under initial pH 12 shock was less than other pH shocks on day 28 (P = 0.01), while no temperature effect was observed on day 28. There was no significant temperature and initial pH effect on cfxA abundance at any time point during incubation, implying that the bacteria that carrying cfxA gene are relatively insensitive to these environmental factors. Overall, directly raising temperature and pH can facilitate pirlimycin removal and decrease mefA and tet(W) relative abundances during storage of manure slurries.202032050366
774810.9971Bacillus subtilis reduces antibiotic resistance genes of animal sludge in vermicomposting by improving heat stress tolerance of Eisenia foetida and bacterial community adjustment. Antibiotic resistance genes (ARGs) in livestock industry have been recognized as a kind of pollutant. The effect of Bacillus subtilis (B. subtilis) as an additive for the reduction of ARGs in animal sludge from livestock and poultry wastewater treatment plant during vermicomposting was investigated. We also evaluated the oxidative stress level and growth of earthworms, Eisenia foetida, bacterial community succession, and the quality of the end products. Two treatments were conducted using B. subtilis, one at 18 °C and another at 28 °C. Controls were setup without the bacteria. The results showed that inoculation of B. subtilis promoted the degradation of organics at 28 °C and increased the germination index to 236%. The increased activities of the superoxide dismutase (1.69 U/mg pr) and catalase (8.05 U/mg pr) and the decreased activity of malondialdehyde (0.02 nmol/mg pr) by B. subtilis at 28 °C showed that the earthworms were relieved of heat stress. The addition of B. subtilis reduced the abundance of 32 target ARGs, including integron (intI-1), transposase (IS613) and resistant genes, such as sulfonamide (sul2), quinolone (oprJ), macrolide-lincosamide-streptogramin group B (ermF, ermB), tetracycline (tetL-02, tetX), β-lactama (blaOXA10-01) and aminoglycoside [strB, aac(6')-Ib(aka aacA4)-01, aac(6')-Ib(aka aacA4)-02]. Organic matter degrading Membranicola, Paludisphaera, Sphingorhabdus and uncultured bacterium belonging to the order Chitinophagales, nitrifying and nitrogen-fixing Singulisphaera and Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, soil remediating Achromobacter, and plant growth promoting Kaistia, Galbibacter and Ilumatobacter were increased significantly (P < 0.05). However, the growth of harmful bacteria such as Burkholderiaceae was inhibited in the vermicompost. In earthworm guts, the probiotic Mesorhizobium was promoted, while the pathogenic uncultured bacterium belonging to the family Enterobacteriaceae was reduced. Besides, B. subtilis enhanced the host relationships between bacteria and ARGs. These findings might be helpful in the removal of ARGs in animal wastes and in understanding the synergy between earthworms and microorganisms.202336529325
874120.9970Acclimation of electroactive biofilms under different operating conditions: comprehensive analysis from architecture, composition, and metabolic activity. Electroactive biofilms (EABs) have aroused wide concern in waste treatment due to their unique capability of extracellular electron transfer with solid materials. The combined effect of different operating conditions on the formation, microbial architecture, composition, and metabolic activity of EABs is still unknown. In this study, the impact of three different factors (anode electrode, substrate concentration, and resistance) on the acclimation and performance of EABs was investigated. The results showed that the shortest start-up time of 127.3 h and highest power density of 0.84 W m(-2) were obtained with carbon brush as electrode, low concentration of substrate (1.0 g L(-1)), and 1000 Ω external resistance (denoted as N1). The EABs under N1 condition also represented strongest redox capacity, lowest internal resistance, and close arrangement of bacteria. Moreover, the EABs cultured under different conditions both showed similar results, with direct electron transfer (DET) dominated from EABs to anode. Microbial community compositions indicated that EABs under N1 condition have lowest diversity and highest abundance of electroactive bacteria (46.68%). Higher substrate concentration (3.0 g L(-1)) promoted the proliferation of some other bacteria without electroactivity, which was adverse to EABs. The metabolic analysis showed the difference of genes related to electron transfer (cytochrome C and pili) and biofilm formation (xap) of EABs under different conditions, which further demonstrated the higher electroactivity of EABs under N1. These results provided a comprehensive understanding of the effect of different operating conditions on EABs including biofilm formation and electrochemical activity.202337749470
776830.9970Drinking water biofiltration: Behaviour of antibiotic resistance genes and the association with bacterial community. Antibiotic resistance genes (ARGs) are being detected in drinking water frequently, constituting a major public health issue. As a typical drinking water treatment process, the biofilter may harbour various ARGs due to the filter biofilms established during the filtration process. The objective of this study was to investigate the behaviour of ARGs (bla(CTX-M), bla(OXA-1), bla(TEM), ermB, tetA, tetG, tetQ, tetW, tetX, sul 1, sul 2, dfrA1 and dfrA12) and their possible association with bacteria in a bench-scale biofiltration system. The impact of filter media on horizontal gene transfer (HGT) was also explored using a model conjugative plasmid, RP1. The biofiltration system comprised four types of biofilters, including sand, granular activated carbon (GAC), GAC sandwich, and anthracite-sand biofilters. Results showed that although the absolute abundance of ARGs decreased (0.97-log reduction on average), the ARGs' abundance normalised to bacterial numbers showed an increasing trend in the filtered water. Biofilms collected from the surface layer revealed the lowest relative abundance of ARGs (p < 0.01) compared to the deeper layer biofilms, indicating that the proportion of ARG-carrying bacteria was greater in the lower position. Most chosen ARG numbers correlated to Proteobacteria, Acidobacteria and Nitrospirae phyla, which accounted for 51.9%, 5.2% and 2.0% of the biofilm communities, respectively. GAC media revealed the highest transfer frequency (2.60 × 10(-5)), followed by anthracite (5.31 × 10(-6)) and sand (2.47 × 10(-6)). Backwashing can reduce the transferability of RP1 plasmid significantly in biofilms but introduces more transconjugants into the planktonic phase. Overall, the results of this study could enhance our understanding of the prevalence of ARGs in drinking water biofiltration treatment.202032650149
799740.9970Survival of Antibiotic Resistant Bacteria and Horizontal Gene Transfer Control Antibiotic Resistance Gene Content in Anaerobic Digesters. Understanding fate of antibiotic resistant bacteria (ARB) vs. their antibiotic resistance genes (ARGs) during wastewater sludge treatment is critical in order to reduce the spread of antibiotic resistance through process optimization. Here, we spiked high concentrations of tetracycline-resistant bacteria, isolated from mesophilic (Iso M1-1-a Pseudomonas sp.) and thermophilic (Iso T10-a Bacillus sp.) anaerobic digested sludge, into batch digesters and monitored their fate by plate counts and quantitative polymerase chain reaction (QPCR) of their corresponding tetracycline ARGs. In batch studies, spiked ARB plate counts returned to baseline (thermophilic) or 1-log above baseline (mesophilic) while levels of the ARG present in the spiked isolate [tet(G)] remained high in mesophilic batch reactors. To compare results under semi-continuous flow conditions with natural influent variation, tet(O), tet(W), and sul1 ARGs, along with the intI1 integrase gene, were monitored over a 9-month period in the raw feed sludge and effluent sludge of lab-scale thermophilic and mesophilic anaerobic digesters. sul1 and intI1 in mesophilic and thermophilic digesters correlated positively (Spearman rho = 0.457-0.829, P < 0.05) with the raw feed sludge. There was no correlation in tet(O) or tet(W) ratios in raw sludge and mesophilic digested sludge or thermophilic digested sludge (Spearman rho = 0.130-0.486, P = 0.075-0.612). However, in the thermophilic digester, the tet(O) and tet(W) ratios remained consistently low over the entire monitoring period. We conclude that the influent sludge microbial composition can influence the ARG content of a digester, apparently as a result of differential survival or death of ARBs or horizontal gene transfer of genes between raw sludge ARBs and the digester microbial community. Notably, mesophilic digestion was more susceptible to ARG intrusion than thermophilic digestion, which may be attributed to a higher rate of ARB survival and/or horizontal gene transfer between raw sludge bacteria and the digester microbial community.201627014196
803450.9970Adding a complex microbial agent twice to the composting of laying-hen manure promoted doxycycline degradation with a low risk on spreading tetracycline resistance genes. Poultry manure is a reservoir for antibiotics and antibiotic resistance genes and composting is an effective biological treatment for manure. This study explored the effect of using two methods of adding a complex microbial agent to the composting of laying-hen manure on doxycycline degradation and tetracycline resistance genes elimination. The results showed that incorporating a complex microbial agent at 0.8% (w/w) on the 0(th) and 11th day (group MT2) effectively degraded doxycycline with a final degradation rate of 46.83 ± 0.55%. The half-life of doxycycline in this group was 21.90 ± 0.00 days and was significantly lower than that of group MT1 (1.6% (w/w) complex microbial agent added on the 0(th) day) and group DT (compost without complex microbial agent). But there was no significant difference in the final degradation rate of doxycycline between group DT and group MT1. The addictive with the complex microbial agent changed the microbial community structure. Bacteroidetes, Firmicutes and Proteobacteria were the dominant phyla during composting. Aerococcus, Desemzia, Facklamia, Lactobacillus, Streptococcus, and Trichococcus were the bacteria related to the degradation of doxycycline. Moreover, the incorporation of a complex microbial agent could decrease the risk on spreading tetracycline resistance genes. The single addition promoted the elimination of tetM, whose possible hosts were Enterococcus, Lactobacillus, Staphylococcus, and Trichococcus. Adding the complex microbial agent twice promoted the elimination of tetX, which was related to the low abundance of Chryseobacterium, Flavobacterium and Neptunomonas in group MT2. Redundancy analysis showed that the bacterial community, residual doxycycline and physiochemical properties have a potential effect on the variation in tetracycline resistance genes levels. Overall, adding the complex microbial agent twice is an effective measure to degrade doxycycline.202032806409
721260.9969Simulated Winter Incubation of Soil With Swine Manure Differentially Affects Multiple Antimicrobial Resistance Elements. Gastrointestinal bacteria that harbor antibiotic resistance genes (ARG) become enriched with antibiotic use. Livestock manure application to cropland for soil fertility presents a concern that ARG and bacteria may proliferate and be transported in the environment. In the United States, manure applications typically occur during autumn with slow mineralization until spring planting season. A laboratory soil incubation study was conducted mimicking autumn swine manure application to soils with concentrations of selected ARG monitored during simulated 120-day winter incubation with multiple freeze-thaw events. Additionally, the effects of two soil moistures [10 and 30% water holding capacity (WHC)] and two manure treatments [raw versus hydrated lime alkaline stabilization (HLAS)] were assessed. Fourteen tetracycline resistance genes were evaluated; tet(D), tet(G), and tet(L) were detected in background soil while swine manure contained tet(A), tet(B), tet(C), tet(G), tet(M), tet(O), tet(Q), and tet(X). By day 120, the manure-borne tet(M) and tet(O) were still detected while tet(C), tet(D), tet(L), and tet(X) genes were detected less frequently. Other tet resistance genes were detected rarely, if at all. The sum of unique tet resistance genes among all treatments decreased during the incubation from an average of 8.9 to 3.8 unique tet resistance genes. Four resistance elements, intI1, bla (ctx-m-32), sul(I), erm(B), and 16s rRNA genes were measured using quantitative PCR. ARG abundances relative to 16S abundance were initially greater in the raw manure compared to background soil (-1.53 to -3.92 log abundance in manure; -4.02 to <-6.7 log abundance in soil). In the mixed manure/soil, relative abundance of the four resistance elements decreased (0.87 to 1.94 log abundance) during the incubation largely because 16S rRNA genes increased by 1.21 log abundance. Throughout the incubation, the abundance of intI1, bla (ctx-m-32), sul(I), and erm(B) per gram in soil amended with HLAS-treated manure was lower than in soil amended with raw manure. Under low initial soil moisture conditions, HLAS treatment reduced the abundance of intI1 and resulted in loss of bla (ctx-m-32), sul(I), and erm(B)] compared to other treatment-moisture combinations. Although one might expect antibiotic resistance to be relatively unchanged after simulated winter manure application to soil, a variety of changes in diversity and relative abundance can be expected.202033391241
718170.9969Effects of UVC doses on the removal of antimicrobial resistance elements from secondary treated sewage. Wastewater treatment plants (WWTPs) currently face major challenges toward the removal of microcontaminants and/or microbial matrices and consequently play an important role in the potential dissemination of biological resistance in freshwater. The ultraviolet (UV) system is a tertiary treatment strategy increasingly applied worldwide, although many studies have shown that disinfected effluent can still contain antibiotic-resistant bacteria and resistance genes. Therefore, to better understand the effects of UV radiation doses on the removal of all resistance elements (antibiotics, antibiotic-resistant bacteria, and antibiotic resistance genes), the present study was designed using a pilot-scale photoreactor. The UV doses could be varied to investigate whether there is an optimal UV dose capable of removing all resistance elements and also if the UV dose frequently applied in full-scale systems is able to reduce the resistance elements. The effect of different UV doses (A, 0-10 mJ/cm(2); B, 10-15 mJ/cm(2); and C, > 15 mJ/cm(2)) in a pilot-scale photoreactor on the removal of antibiotics, antibiotic-resistant bacteria, and genes from the effluent of a UASB reactor followed by a biological trickling filter system (UASB-TF) fed with real sanitary sewage was investigated. Samples of influent and effluent from the UVC photoreactor were collected, and the concentration levels of norfloxacin (NOR), ciprofloxacin (CIP), and levofloxacin (LEV) were assessed. The qnrB, sul1, ermB, integron-integrase (intI1), and 16S rRNA genes, total heterotrophic bacteria (THB), and bacterial resistance to azithromycin and sulfamethoxazole were also investigated. Results indicated that LEV and intI1 were found in the highest median concentrations in the photoreactor influent. Although most antibiotics (NOR and CIP) and ARGs (intI1, 16S rRNA, and qnrB) were apparently better removed with the highest UV dose (> 15 mJ/cm(2)) applied, except for LEV, sul1, and ermB genes, the Kruskal-Wallis test reported no significant difference between low and high doses. ARB removal (from 80 to 100%) was observed at all UV doses. Principal component analysis (PCA) suggested a clear pattern of pollutant groups, i.e., antibiotics, ARG, and ARB, which exhibited low (median of 8-16%), medium (37-96%), and high (> 97%) removal percentages, respectively. These results demonstrated that UVC photoreactors can be an alternative to complement biological treatment in sewage treatment plants at the dose normally applied in full-scale WWTPs (> 15 mJ/cm(2)). However, there was no optimal single dose capable of removing all the resistance elements investigated.202539873874
775080.9969Efficient removal of enrofloxacin in swine wastewater using eukaryotic-bacterial symbiotic membraneless bioelectrochemical system. A eukaryotic-bacterial symbiotic membraneless bioelectrochemical system (EBES) reactor with eukaryotic-bacteria symbiotic cathode was developed to treat swine wastewater containing enrofloxacin (ENR), which had high performance at ENR tolerance and operational stability. With ENR concentrations shifting from 2 to 50 mg/L, the removal efficiencies of ENR, chemical oxygen demand (COD) and NH(4)(+)-N always were higher than 95 %, and the maximum power output (≥343 mW/m(3)) could be achieved. At 20 mg/L ENR, the removal efficiencies of ENR, COD and NH(4)(+)-N respectively reached to 99.4 ± 0.1 %, 98.5 % ± 0.1 %, and 96.3 % ± 0.5 %, corresponding to the open circuit voltage and maximum power density (P(max)) of EBES were 851 mV and 455 mW/m(3). The community analyses showed that bacteria (Comamonas, Rhodobacter, Rhodococcus, and Vermiphilaceae et al.), algae (Chlorella) and fungi (Rozellomycota, Trebouxiophyceae, Exophiala, and Aspergillus et al.) at genus level were the dominate populations in the EBES, and their abundance increased with ENR concentration, suggesting they played key roles to remove ENR and another nutrient element. The low relative abundances (1.9 ×10(-7) to 1.1 ×10(-5) copies/g) of aac (6')-ib-cr, qnrA, qnrD, qnrS, and gyrA in effluent revealed that the present EBES reactor had superior capabilities in controlling antibiotic-resistance genes and antibiotic-resistant bacteria. Our trial experiments provided a novel way for antibiotic livestock wastewater treatment.202539938376
528590.9969Antibiotic Use in Beekeeping: Implications for Health and Environment from a One-Health Perspective. BACKGROUND: The use of antibiotics in beekeeping has potential implications for honeybee health and environmental contamination. Recent research indicates that extensive antibiotic use in beekeeping, especially oxytetracycline, promotes antimicrobial resistance in bee-related bacteria. Honeybees can transport oxytetracycline-resistance genes during foraging, potentially establishing reservoirs of resistance in the colony and facilitating intergeneric gene transfer among various gut bacteria as well as in the microbiome of the flowers and the wider environment, where honeybees can spread antibiotic-resistance genes over a large distance. This study investigates the effects of oxytetracycline hydrochloride (OTC) treatment on honeybees from a One Health perspective, examining antibiotic residues in honey, environmental spread, and the presence of tetracycline-resistance genes (TET-RGs). METHODS: In the spring of 2022, two groups of four honeybee hives were placed near an almond grove in Central Italy. One group was treated with 1.68 g of OTC, while the other remained untreated. Samples were collected from bees, honey, hive entrances, and flowers before treatment and at 3 as well as 9 days post-treatment. OTC residues and TET-RGs were analyzed to assess contamination and resistance gene dissemination. RESULTS: OTC residues were detected in honey from both treated (day 3: 263,250.0 ± 100,854.3 µg/kg; day 9: 132,600 ± 146,753.9 µg/kg) and untreated hives (day 3: 20.5 ± 8.2 µg/kg; day 9: 135.8 ± 198.6 µg/kg), suggesting cross-contamination. Residues were also found in almond tree flowers (0.7 ± 0.1 µg/kg), with TET-RGs (tet(K), tet(L), tet(M), tet(B), tet(O), tet(D)) detected pre- and post-treatment. In honeybee gut bacteria, resistance genes (tet(M), tet(A), tet(D), tet(B)) appeared post-treatment in both groups. No significant correlation was observed between hive distance and resistance gene presence in flowers, although the presence of other farms located within the bees' flight range, in which OTC might have been used in the past, could have influenced the results. CONCLUSIONS: These findings highlight the risk of OTC-induced antibiotic cross-contamination and the spread of TET-RG, raising concerns for bee health and environmental safety. Given honeybees' social nature and the negative effects of antibiotics on their health, an antibiotic-free management approach is recommended for sustainable apiculture.202540298498
7761100.9969Fate and removal of bacteria and antibiotic resistance genes in horizontal subsurface constructed wetlands: Effect of mixed vegetation and substrate type. This study aimed to investigate the influence of cropping method and substrate type on the fate and the removal of bacterial and antibiotic resistance genes (ARGs) indicators from primary wastewater by constructed wetlands (CWs) during startup and maturation stages. Four small-scale CWs differing in their plantation pattern (monoculture vs. polyculture) and substrate type were constructed and operated under field conditions. While for bacteria, the greatest impact of the cropping method and substrate type on removal was during the startup stage rather than the maturation stage, for ARGs, such impact was significant at both stages. During startup, the removal efficiencies of heterotrophic bacteria, fecal coliforms, E. coli, 16S rRNA genes and lacZ increased with the operation time. At maturation, the removal efficiencies were constant and were within the range of 89.2-99.4%, 93.7-98.9%, 89-98.8%, 94.1-99.6% and 92.9-98.7%, respectively. The removal efficiencies of intl1, tetM, intl1, sul1, ermB and total ARGs were also increased with the operation time. However, they were ARG type and configuration-dependent; at maturation they ranged between 50.7%-89.4%, 85.9%-97%, 49.6%-92.9%, 58.2%-96.7% and 79.9-94.3%, respectively. The tuff-filled serially planted CW was also the only one capable of removing these genes at similar high efficiency. Metagenomic analysis showed that none of the ARGs was among the most common ARGs in water and biofilm samples; rather most ARGs belonged to bacterial efflux transporter superfamilies. Although ARGs were removed, they were still detected in substrate biofilm and their relative concentrations were increased in the effluents. While the removal of both bacteria and ARGs was higher during summer compared to winter, the season had no effect on the removal pattern of ARGs. Hence, combination of the serial plantation with substrate having high surface area is a potential strategy that can be used to improve the performance of CWs.202133338689
7996110.9969A sludge bulking wastewater treatment plant with an oxidation ditch-denitrification filter in a cold region: bacterial community composition and antibiotic resistance genes. Bacterial community structure of activated sludge directly affects the stable operation of WWTPS, and these bacterial communities may carry a variety of antibiotic resistance genes (ARGs), which is a threat to the public health. This study employed 16S rRNA gene sequencing and metagenomic sequencing to investigate the bacterial community composition and the ARGs in a sludge bulking oxidation ditch-denitrification filter WWTP in a cold region. The results showed that Trichococcus (20.34%), Blautia (7.72%), and Faecalibacterium (3.64%) were the main bacterial genera in the influent. The relative abundances of norank_f_Saprospiraceae and Candidatus_Microthrix reached 10.24% and 8.40%, respectively, in bulking sludge, and those of norank_f_Saprospiraceae and Candidatus_Microthrix decreased to 6.56 and 7.10% after the anaerobic tank, indicating that the anaerobic tank had an inhibitory effect on filamentous bacteria. After 20 mJ/cm(2) UV disinfection, about 540 bacterial genera, such as Romboutsia (7.99%), Rhodoferax (7.98%), and Thermomonas (4.13%), could still be detected in the effluent. The ARGs were 345.11 ppm in the influent and 11.20 ppm in the effluent; 17 subtypes, such as sul1, msrE, aadA5, ErmF, and tet(A), could be detected throughout the entire process. These ARG subtypes were persistent ARGs with a high health risk. Network analysis indicated that the changes in filamentous bacteria norank_f_Saprospiraceae abundance mainly contributed to the abundance shift of MexB, and Acinetobacter mainly increased the abundance of drfA1. These results above will provide theoretical support for the sludge bulking and ARGs controls of WWTPs in cold regions.202336495431
8046120.9969Responses of aerobic granular sludge to fluoroquinolones: Microbial community variations, and antibiotic resistance genes. In this study, aerobic granular sludge (AGS) was operated under high levels of ammonium for removing three fluoroquinolones (FQs), i.e., ciprofloxacin (CFX), ofloxacin (OFX), and norfloxacin (NFX) at 3, 300, and 900 µg/L, respectively. Two key objectives were to investigate the differential distribution of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) in sludge fractions and to evaluate correlations between ARGs and MGEs to nitrifying and denitrifying bacteria. AGS showed excellent stability under the exposure of FQs, with nitrite-oxidizing bacteria (NOB) more sensitive to FQs than ammonium-oxidizing bacteria (AOB). Specific oxygen utilization rates (SOUR) showed a reduction of 26.9% for NOB but only 4.0% of the reduced activity of AOB by 3 μg/L FQs. AGS performed better removal efficiencies for CFX and NFX than OFX, and the efficiencies increased with their elevated concentrations, except at 900 μg/L FQs. The elevated FQ concentrations led to a significant enrichment of intI1 and genus Thauera, while qnrD and qnrS showed no accumulation. Compared to nitrifiers, FQs relevant ARGs and the intI1 gene preferred to exist in denitrifiers, and the abundance of denitrifiers behaved a decreasing trend with the sludge size. Two quinoline-degrading bacteria were found in the AGS system, i.e., Alicycliphilus and Brevundimonas, possibly carrying qnrS and qnrD, respectively. Their relative abundance increased with the sludge size, which was 2.18% in sludge <0.5 mm and increased to 3.70% in sludge >2.0 mm, suggesting that the AGS may be a good choice in treating FQs-containing wastewater.202133676249
7793130.9969Treatment of pharmaceutical wastewater by ionizing radiation: Removal of antibiotics, antimicrobial resistance genes and antimicrobial activity. In present study, the treatment of real pharmaceutical wastewater from an erythromycin (ERY) production factory by gamma irradiation was investigated. Results showed that a variety of antimicrobial resistance genes (ARGs), involving MLSB, tet, bla, multidrug, sul, MGEs and van genes and plentiful 9 bacterial phyla were identified in the raw wastewater. In addition to ERY, sulfamethoxazole (SMX) and tetracycline (TC) were also identified with the concentration of 3 order of magnitude lower than ERY. Results showed that the abatement of ARGs and antibiotics was much higher than that of antimicrobial activity and COD. With the absorbed dose of 50 kGy, the removal percentage of ARGs, ERY, antimicrobial activity and COD was 96.5-99.8%, 90.0%, 47.8% and 10.3%, respectively. The culturable bacteria were abated fast and completely at 5.0 kGy during gamma irradiation. The genus Pseudomonas was predominant in raw wastewater (56.7%) and its relative abundance decreased after gamma irradiation, to 1.3% at 50 kGy. With addition of peroxymonosulfate (PMS, 50 mM), the antimicrobial activity disappeared completely and ERY removal reached as high as 99.2% at the lower absorbed dose of 25 kGy. Ionizing radiation-coupled technique is a potential option to treat pharmaceutical wastewater for reduction of antibiotics, ARGs and antimicrobial activity.202134088196
7951140.9968Proliferation of antibiotic resistance genes in microbial consortia of sequencing batch reactors (SBRs) upon exposure to trace erythromycin or erythromycin-H2O. A variety of antibiotics and their metabolites at sub-inhibitory level concentrations are suspected to expand resistance genes in the environment. However, knowledge is limited on the causal correlation of trace antibiotics or their metabolites with resistance proliferation. In this study, erythromycin (ERY) resistance genes were screened on microbial consortia of sequencing batch reactors (SBRs) after one year acclimation to ERY (100 μg/L) or dehydrated erythromycin (ERY-H(2)O, 50 μg/L). The identified esterase gene ereA explains that ERY could be degraded to six products by microbes acclimated to ERY (100 μg/L). However, ERY could not be degraded by microbes acclimated to ERY-H(2)O (50 μg/L), which may be due to the less proliferated ereA gene. Biodegradation of ERY required the presence of exogenous carbon source (e.g., glucose) and nutrients (e.g., nitrogen, phosphorus) for assimilation, but overdosed ammonium-N (>40 mg/L) inhibited degradation of ERY. Zoogloea, a kind of biofilm formation bacteria, became predominant in the ERY degradation consortia, suggesting that the input of ERY could induce biofilm resistance to antibiotics. Our study highlights that lower μg/L level of ERY or ERY-H(2)O in the environment encourages expansion of resistance genes in microbes.201121482429
8036150.9968Abundances of Tetracycline Resistance Genes and Tetracycline Antibiotics during Anaerobic Digestion of Swine Waste. The impact of anaerobic digestion of animal waste on the persistence of antibiotic resistance genes (ARGs) and antibiotics is not widely studied. Two identical, 800-L digesters seeded with swine slurry were followed up to 100 d in three separate trials. The trials received varying amounts of antibiotic-free corn ( L.) mixed with water to maintain the digestion process. Biogas production, seven tetracycline resistance () genes, and three tetracyclines and their transformation products were measured. Biogas production proportionally increased as the feeding loads increased between trials. In Trial 1, log gene copies showed small but statistically significant ( < 0.01) increases during digestion. In Trial 2, anaerobic digestion did not have a significant ( > 0.05) effect except for significant reductions in B ( < 0.0001) and G ( = 0.0335) log gene copies. In Trial 3, which received the highest amount of corn mix, log copies of the 16S ribosomal RNA and the genes significantly ( < 0.0001) reduced over time during digestion. Up to 36 μg L tetracycline, 112 μg L chlortetracycline, 11.9 mg L isochlortetracycline, and 30 μg L 4-epitetracycline were detected both in the liquid and solid digestates. Results of this study revealed that although anaerobic digestion of swine waste can produce useful biogas, it does not result in complete removal of bacteria, ARGs, and antibiotics regardless of differences in the feeding loads between trials. Further effluent and sludge treatments are required prior to their downstream use in crop production to minimize emergence and environmental dissemination of antimicrobial-resistant bacteria through animal manure.201930640349
7775160.9968Accumulation of pharmaceuticals, Enterococcus, and resistance genes in soils irrigated with wastewater for zero to 100 years in central Mexico. Irrigation with wastewater releases pharmaceuticals, pathogenic bacteria, and resistance genes, but little is known about the accumulation of these contaminants in the environment when wastewater is applied for decades. We sampled a chronosequence of soils that were variously irrigated with wastewater from zero up to 100 years in the Mezquital Valley, Mexico, and investigated the accumulation of ciprofloxacin, enrofloxacin, sulfamethoxazole, trimethoprim, clarithromycin, carbamazepine, bezafibrate, naproxen, diclofenac, as well as the occurrence of Enterococcus spp., and sul and qnr resistance genes. Total concentrations of ciprofloxacin, sulfamethoxazole, and carbamazepine increased with irrigation duration reaching 95% of their upper limit of 1.4 µg/kg (ciprofloxacin), 4.3 µg/kg (sulfamethoxazole), and 5.4 µg/kg (carbamazepine) in soils irrigated for 19-28 years. Accumulation was soil-type-specific, with largest accumulation rates in Leptosols and no time-trend in Vertisols. Acidic pharmaceuticals (diclofenac, naproxen, bezafibrate) were not retained and thus did not accumulate in soils. We did not detect qnrA genes, but qnrS and qnrB genes were found in two of the irrigated soils. Relative concentrations of sul1 genes in irrigated soils were two orders of magnitude larger (3.15 × 10(-3) ± 0.22 × 10(-3) copies/16S rDNA) than in non-irrigated soils (4.35 × 10(-5)± 1.00 × 10(-5) copies/16S rDNA), while those of sul2 exceeded the ones in non-irrigated soils still by a factor of 22 (6.61 × 10(-4) ± 0.59 × 10(-4) versus 2.99 × 10(-5) ± 0.26 × 10(-5) copies/16S rDNA). Absolute numbers of sul genes continued to increase with prolonging irrigation together with Enterococcus spp. 23S rDNA and total 16S rDNA contents. Increasing total concentrations of antibiotics in soil are not accompanied by increasing relative abundances of resistance genes. Nevertheless, wastewater irrigation enlarges the absolute concentration of resistance genes in soils due to a long-term increase in total microbial biomass.201223049795
8045170.9968Correlation among extracellular polymeric substances, tetracycline resistant bacteria and tetracycline resistance genes under trace tetracycline. Antibiotic resistance occurrences and proliferation in activated sludge have attracted more and more attention nowadays. However, the role which extracellular polymeric substance (EPS) plays on the antibiotic resistance is not clear. The changes and correlation among EPS, tetracycline (TC) resistant bacteria (TRB) and TC resistance genes (TRGs) of sequencing batch reactors (SBRs) were investigated. Performance of SBR without TC was compared with two other SBRs to which different amounts of TC were added. Total average EPS contents were found to increase significantly from 66 mg g−1 VSS to 181 mg g−1 VSS as the TC concentrations increased from 0 to 100 μg L−1. As the EPS content increased, TRB in sludge of the three SBRs increased significantly from 105 to 106 colony forming unit mL−1 after being exposed to TC. In addition, the concentrations of three groups of TRGs (copies mL−1) were determined by real-time fluorescence quantitative polymerase chain reaction and followed the order: efflux pump genes > ribosome protected genes > degradation enzyme genes. The numbers of TRGs in the idle stage were larger than those in the aeration sludge. Correlation coefficients (R2) between EPS and TRB in sludge were 0.823 (p < 0.01) while the correlation between EPS and total TRGs was poor (R2 = 0.463, p > 0.05). But it showed the same tendency that EPS and TRGs in sludge increased with the increasing of TC.201425461932
8047180.9968Simultaneous elimination of antibiotics and antibiotics resistance genes in nitritation of source-separated urine. Antibiotics in human urine could accelerate dissemination of antibiotics resistance genes (ARGs), posing potential threat to sewage. The nitritation of source-separated urine was a critical step to realize the urine resourcelization and nitrogen stabilization. However, the synergic control on antibiotics and ARGs during urine nitritation was unrevealed. This study investigated the removal profiles of five typical antibiotics and the shifts of microbial community and ARGs during stable nitritation. The result showed that sulfamethoxazole and roxithromycin were effectively eliminated with high removal efficiency of (95 ± 5) % and (90 ± 10) %, followed by enrofloxacin with removal efficiency of (60 ± 5) %, whereas trimethoprim and chloramphenicol showed low removal efficiency of less than 40 %. Ammonia oxidation bacteria and heterotrophic bacteria equally contributed to elimination of sulfamethoxazole with a high biodegradation rate of 0.1534 L/gVSS·h, while sorption and biodegradation jointly promoted other antibiotics removal. The total relative abundance of top 25 bacteria genera was decreased by 10 %. The total relative abundance of top 30 ARGs was decreased by more than 20 %, which was corresponding to the variation of bacterial community. The findings in this research would get a deeper insight into the eliminating antibiotics and controlling ARGs dissemination during nitritation of source-separated urine.202235897182
7962190.9968Linking the Effect of Antibiotics on Partial-Nitritation Biofilters: Performance, Microbial Communities and Microbial Activities. The emergence and spread of antibiotics resistance in wastewater treatment systems have been pointed as a major environmental health problem. Nevertheless, research about adaptation and antibiotics resistance gain in wastewater treatment systems subjected to antibiotics has not been successfully developed considering bioreactor performance, microbial community dynamics and microbial activity dynamics at the same time. To observe this in autotrophic nitrogen removal systems, a partial-nitritation biofilter was subjected to a continuous loading of antibiotics mix of azithromycin, norfloxacin, trimethoprim, and sulfamethoxazole. The effect of the antibiotics mix over the performance, bacterial communities and bacterial activity in the system was evaluated. The addition of antibiotics caused a drop of ammonium oxidation efficiency (from 50 to 5%) and of biomass concentration in the bioreactor, which was coupled to the loss of ammonium oxidizing bacteria Nitrosomonas in the bacterial community from 40 to 3%. Biomass in the partial nitritation biofilter experienced a sharp decrease of about 80% due to antibiotics loading, but the biomass adapted and experienced a growth by stabilization under antibiotics feeding. During the experiment several bacterial genera appeared, such as Alcaligenes, Paracoccus, and Acidovorax, clearly dominating the bacterial community with >20% relative abundance. The system reached around 30% ammonium oxidation efficiency after adaptation to antibiotics, but no effluent nitrite was found, suggesting that dominant antibiotics-resistant phylotypes could be involved in nitrification-denitrification metabolisms. The activity of ammonium oxidation measured as amoA and hao gene expression dropped a 98.25% and 99.21%, respectively, comparing the system before and after the addition of antibiotics. On the other hand, denitrifying activity increased as observed by higher expression of nir and nos genes (83.14% and 252.54%, respectively). In addition, heterotrophic nitrification cyt c-551 was active only after the antibiotics addition. Resistance to the antibiotics was presumably given by ermF, carA and msrA for azithromycin, mutations of the gyrA and grlB for norfloxacin, and by sul123 genes for sulfamethoxazole. Joined physicochemical and microbiological characterization of the system were used to investigate the effect of the antibiotics over the bioprocess. Despite the antibiotics resistance, activity of Bacteria decreased while the activity of Archaea and Fungi increased.201829535704