The response of copper resistance genes, antibiotic resistance genes, and intl1/2 to copper addition during anaerobic digestion in laboratory. - Related Documents




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719701.0000The response of copper resistance genes, antibiotic resistance genes, and intl1/2 to copper addition during anaerobic digestion in laboratory. Heavy metal pollution can serve as a selective pressure for antibiotic resistance genes in polluted environments. Anaerobic fermentation, as a recommended wastewater treatment method, is an effective mitigation measure of antibiotic resistance diffusion. To explore the influence of copper on anaerobic fermentation, we exposed the fermentation substrate to copper in a laboratory setup. We found that the relative abundance of 8 genes (pcoD, tetT, tetA, tetB, tetO, qnrS, ermA and ermB) increased at the late stage of fermentation and their abundance was linked to copper content. Corynebacterium and Streptococcus were significantly positively correlated with ermA, ermB, tetA and tetB (P < 0.05). The relative abundance of tetT was significantly positively correlated with Terrisporobacter, Clostridium_sensu_stricto_1 and Turicibacter (P < 0.05). We screened 90 strains of copper resistant bacteria from blank, medium and high copper test groups on days 25, 31 and 37. The number of fragments carried by a single strain increased with time while intl1, ermA and ermB existed in almost all combinations of the multiple fragments we identified. The relative abundance of these three genes were linearly correlated with Corynebacterium and Streptococcus. The antibiotic resistance genes carried by class 1 integrons gradually increased with time in the fermentation system and integrons carrying ermA and ermB most likely contributed to host survival through the late stages of fermentation. The genera Corynebacterium and Streptococcus may be the primary carriers of such integrated mobile gene element and this was most likely the reason for their rebound in relative abundance during the late fermentation stages.202133418156
343210.9999Insights into the amplification of bacterial resistance to erythromycin in activated sludge. Wastewater treatment plants are significant reservoirs for antimicrobial resistance. However, little is known about wastewater treatment effects on the variation of antibiotic resistance. The shifts of bacterial resistance to erythromycin, a macrolide widely used in human medicine, on a lab-scale activated sludge system fed with real wastewater was investigated from levels of bacteria, community and genes, in this study. The resistance variation of total heterotrophic bacteria was studied during the biological treatment process, based on culture dependent method. The alterations of bacterial community resistant to erythromycin and nine typical erythromycin resistance genes were explored with molecular approaches, including high-throughput sequencing and quantitative polymerase chain reaction. The results revealed that the total heterotrophs tolerance level to erythromycin concentrations (higher than 32 mg/L) was significantly amplified during the activated sludge treatment, with the prevalence increased from 9.6% to 21.8%. High-throughput sequencing results demonstrated an obvious increase of the total heterotrophic bacterial diversity resistant to erythromycin. Proteobacteria and Bacteroidetes were the two dominant phyla in the influent and effluent of the bioreactor. However, the prevalence of Proteobacteria decreased from 76% to 59% while the total phyla number increased greatly from 18 to 29 through activated sludge treatment. The gene proportions of erm(A), mef(E) and erm(D) were greatly amplified after biological treatment. It is proposed that the transfer of antibiotic resistance genes through the variable mixtures of bacteria in the activated sludge might be the reason for the antibiotic resistance amplification. The amplified risk of antibiotic resistance in wastewater treatment needs to be paid more attention.201525957255
712420.9999Changes in diversity of cultured bacteria resistant to erythromycin and tetracycline in swine manure during simulated composting and lagoon storage. This study investigated the impact of composting and lagoon storage on survival and change in diversity of tetracycline-resistant (Tc(r) ) and erythromycin-resistant (Em(r) ) bacteria and the resistance genes they carry in swine manure. Treatments were arranged as a 2 × 2 factorial design: composting vs lagoon storage and 0 vs 1% Surround WP Crop Protectant (a clay product) in three replicates. After 48 days of treatments, resistant bacteria were enumerated by selective plating and identified by 16S rRNA gene sequencing. The erm and the tet gene(s) carried by the resistant isolates were screened using class-specific PCR assays. The plate counts of Tc(r) and Em(r) bacteria decreased by 4-7 logs by composting, but only by 1-2 logs by the lagoon treatment. During the treatments, Acinetobacter gave way to Pseudomonas and Providencia as the largest resistant genera. The clay product had little effect on survival or diversity of resistant bacteria. Of six classes of erm and seven classes of tet genes tested, changes in prevalence were also noted. The results indicate that composting can dramatically shift Tc(r) and Em(r) bacterial populations, and composting can be an effective and practical approach to decrease dissemination of antibiotic resistance from swine farms to the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: The presented research provided evidence that composting is much more effective than lagoon storage in dramatically decreasing culturable bacteria resistant to erythromycin and tetracycline in swine manure. Considerable diversity changes of resistant bacteria were also demonstrated during composting or lagoon storage. Overall, Acinetobacter was the major resistant genus in untreated swine manure, but pseudomonads and Providencia became the major resistant genera after the treatments. This is the first study that investigated diversity changes of cultured bacteria resistant to these two antibiotics during composting and lagoon storage of swine manure. New genes encoding resistance to the two antibiotics were also implied in the cultured isolates.201526031793
712330.9999Presence and fate of antibiotic residues, antibiotic resistance genes and zoonotic bacteria during biological swine manure treatment. The presence and dissemination of antibiotic residues, antibiotic resistance genes and zoonotic bacteria in the environment is of growing concern worldwide. Manure management practices, such as biological removal of nitrogen from swine manure, may help to decrease levels of antibiotic residues, antibiotic resistance genes and zoonotic bacteria present in manure before fertilization, thereby reducing environmental contamination. Therefore, the aim of this study was to monitor the presence and fate of seven antibiotic residues (colistin, sulfadiazine, trimethoprim, doxycycline, oxytetracycline, ceftiofur and tylosin A), nine antibiotic resistance genes (tet(B), tet(L), tet(M), tet(O), tet(Q), tet(W), erm(B), erm(F) and sul2) and two zoonotic bacteria (Salmonella Typhimurium and Campylobacter coli) during biological nitrogen removal from swine manure over time. Samples from the raw manure, the solid fraction, the liquid fraction and the storage lagoon were analyzed on two farms at six time points with an interval of two weeks. Only the antibiotics which were used during the three months preceding the first sampling could be detected before and after biological nitrogen removal from swine manure. Of all the antibiotics studied, doxycycline was recovered in all of the samples and sulfadiazine was recovered in most samples on both farms. For both antibiotics, there appears to be a reduction of the amount of residues present in the storage lagoon compared to the liquid fraction, however, this reduction was not statistically significant. A significant reduction of the relative abundances of most of the antibiotic resistance genes studied was observed when comparing the liquid fraction and the storage lagoon. For tet(L), no differences were observed between the fractions sampled and for sul2 and erm(F), a significant increase in relative abundances was observed on the second farm sampled. For the zoonotic bacteria, a reduction of at least 1 log was observed after biological nitrogen removal from swine manure. The results indicate that the concentration of certain antibiotic residues and several antibiotic resistance genes and the amount of zoonotic bacteria present in the manure may be reduced in the end product of the biological nitrogen removal from swine manure.201930878661
720140.9999Evolution of microbial community and drug resistance during enrichment of tetracycline-degrading bacteria. Recently, the microbial degradation of tetracycline has been widely reported. However, its potential risks in treating wastewater containing high concentrations of tetracycline have not been fully evaluated. In this study, the evolution of the microbial community and drug resistance was traced during the enrichment of tetracycline-degrading bacteria. The results showed that some minor compositions such as Shewanella, Bacillus, and Pseudomonas in the seed sludge became the predominant genera in the enrichment cultures when continuously using tetracycline as the sole carbon source, especially some possible pathogenic bacteria increased significantly in this process. The abundances of most TRGs/16S rDNA were increased after enrichment, although the relative abundance of tetA and tetL genes decreased to some extent. From the enrichment culture, 7 predominant tetracycline-degrading strains were isolated, of which TD-1 (Bacillus) and TD-5 (Shewanella) presented high degradation efficiencies (6-day degradation rate > 95%, half-life period of tetracycline ≈ 24 h). In addition, multiple TRGs, mobile genetic elements (MGEs) and even gene cassettes were found in each tetracycline-degrading isolate. The findings suggested that some risks such as the pathogenicity of isolates and the spread of ARGs should be considered when the biodegradation method is used to treat wastewater polluted with high concentrations of tetracycline.201930660087
719850.9998Abundance and distribution of antibiotic resistance genes in a full-scale anaerobic-aerobic system alternately treating ribostamycin, spiramycin and paromomycin production wastewater. The occurrence of antibiotic-resistant bacteria and antibiotic resistance genes (ARGs) has been intensively investigated for wastewater treatment systems treating single class of antibiotic in recent years. However, the impacts of alternately occurring antibiotics in antibiotic production wastewater on the behavior of ARGs in biological treatment systems were not well understood yet. Herein, techniques including high-capacity quantitative PCR and quantitative PCR (qPCR) were used to investigate the behavior of ARGs in an anaerobic-aerobic full-scale system. The system alternately treated three kinds of antibiotic production wastewater including ribostamycin, spiramycin and paromomycin, which referred to stages 1, 2 and 3. The aminoglycoside ARGs (52.1-79.3%) determined using high-capacity quantitative PCR were the most abundant species in all sludge samples of the three stages. The total relative abundances of macrolide-lincosamide-streptogramin (MLS) resistance genes and aminoglycoside resistance genes measured using qPCR were significantly higher (P < 0.05) in aerobic sludge than in sewage sludge. However, the comparison of ARGs acquired from three alternate stages revealed that MLS genes and the aminoglycoside ARGs did not vary significantly (P > 0.05) in both aerobic and anaerobic sludge samples. In aerobic sludge, one acetyltransferase gene (aacA4) and the other three nucleotidyltransferase genes (aadB, aadA and aadE) exhibited positive correlations with intI1 (r (2) = 0.83-0.94; P < 0.05), implying the significance of horizontal transfer in their proliferation. These results and facts will be helpful to understand the abundance and distribution of ARGs from antibiotic production wastewater treatment systems.201728551881
709260.9998Sulfonamide antibiotics in the Northern Yellow Sea are related to resistant bacteria: implications for antibiotic resistance genes. Antibiotic resistance gene (ARG) residues and the mode of transmission in marine environments remain unclear. The sulfonamide (SAs) concentrations, different genes and total bacterial abundance in seawater and sediment of the Northern Yellow Sea were analyzed. Results showed the genes sul I and sul II were present at relatively high concentrations in all samples, whereas the gene sul III was detected fewer. The ARGs concentrations in the sediment were 10(3) times higher than those in water, which indicated sediment was essential ARG reservoir. Statistical analysis revealed the total antibiotic concentration was positively correlated with the relative abundance of the gene sul I and sul II. The relative abundances of the gene sul I and the gene sul II were also correlated positively with those of the gene int1. This correlation demonstrated that SAs exerted selective pressure on these ARGs, whereas the gene int1 could be implicated in the propagation of the genes sul I and sul II in marine environments.201424928456
720270.9998Cyanobacterial extracellular antibacterial substances could promote the spread of antibiotic resistance: impacts and reasons. Many studies have shown that antibiotic resistance genes (ARGs) can be facilitated by a variety of antibacterial substances. Cyanobacteria are photosynthetic bacteria that are widely distributed in the ocean. Some extracellular substances produced by marine cyanobacteria have been found to possess antibacterial activity. However, the impact of these extracellular substances on ARGs is unclear. Therefore, we established groups of seawater microcosms that contained different concentrations (1000, 100, 10, 1, 0.1, 0.01, and 0 μg mL(-1)) of cyanobacterial extracellular substances (CES), and tracked the changes of 17 types of ARGs, the integron gene (intI1), as well as the bacterial community at different time points. The results showed that CES could enrich most ARGs (15/17) in the initial stage, particularly at low concentrations (10 and 100 μg mL(-1)). The correlation analysis showed a positive correlation between several ARGs and intI1. It is suggested that the abundance of intI1 increased with CES may contribute to the changes of these ARGs, and co-resistance of CES may be the underlying reason for the similar variation pattern of some ARGs. Moreover, the results of qPCR and high-throughput sequencing of 16S rRNA showed that CES had an inhibitory impact on the growth of bacterial communities. High concentrations of CES were found to alter the structure of bacterial communities. Co-occurrence networks showed that bacteria elevated in the high concentration group of CES and might serve as the potential hosts for a variety of ARGs. In general, marine cyanobacteria could play an important role in the global dissemination of ARGs and antibiotic-resistant bacteria (ARBs).202337947439
724780.9998Environmental analysis of typical antibiotic-resistant bacteria and ARGs in farmland soil chronically fertilized with chicken manure. Antibiotics and the corresponding resistant bacteria and resistance genes (ARGs) are generally considered emerging pollutants. To assess the impacts of tetracycline (TC) and sulfonamide (SA) antibiotics that are eliminated with fecaluria as drug prototypes, farmland soil used to research long-term fertilization with chicken manure was collected at four sites in Shandong Province. In this study, the rates of bacterial drug resistance to the same antibiotic decreased with an increase in the concentration of that antibiotic, and the resistance rates to TCs were lower than those to SAs. PCR of ARGs revealed that the ARGs detected at the highest frequency were the TC resistance genes tetW and tetO and the SA resistance genes sul1 and sul2. Real-time qPCR showed that the quantities of ARGs in farmland soil fertilized with chicken manure were significantly greater compared with the control soil. Moreover, significant correlations (R(2)=0.9525, p<0.05) between the number of sul ARGs and the total SA concentration were observed in all of the soil samples. In summary, this study showed that SAs can induce the appearance of ARGs and pollute the soil environment.201728340477
718990.9998Comparative 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
3521100.9998Indirect evidence of transposon-mediated selection of antibiotic resistance genes in aquatic systems at low-level oxytetracycline exposures. Subinhibitory levels of antibiotics can promote the development of antibiotic resistance in bacteria. However, it is unclear whether antibiotic concentrations released into aquatic systems exert adequate pressure to select populations with resistance traits. To examine this issue, 15 mesocosms containing pristine surface water were treated with oxytetracycline (OTC) for 56 days at five levels (0, 5, 20, 50, and 250 microg L(-1)), and six tetracycline-resistance genes (tet(B), tet(L), tet(M), ted(O), tet(Q), and tet(W)), the sum of those genes (tet(R)), "total" 16S-rRNA genes, and transposons (Tn916 and Tn 1545) were monitored using real-time PCR. Absolute water-column resistance-gene abundances did not change at any OTC exposure. However, an increase was observed in the ratio of tet(R) to 16S-rRNA genes in the 250 microg L(-1) OTC units, and an increase in the selection rate of Tc(r) genes (relative to 16S-rRNA genes) was seen when OTC levels were at 20 microg L(-1). Furthermore, tet(M) and Tn916/1545 gene abundances correlated among all treatments (r2 = 0.701, p = 0.05), and there were similar selection patterns of tetR and Tn916/1545 genes relative to the OTC level, suggesting a possible mechanism for retention of specific resistance genes within the systems.200818754392
8031110.9998Anaerobic Digestion of Tetracycline Spiked Livestock Manure and Poultry Litter Increased the Abundances of Antibiotic and Heavy Metal Resistance Genes. Anaerobic digestion is used for the treatment of animal manure by generating biogas. Heavy metals cause environmental pollutions and co-select for antimicrobial resistance. We evaluated the impact of mesophilic anaerobic digestion of cattle manure (CM), swine manure (SM) and poultry litter (PL) on the concentrations of seven tetracycline [tet(A), tet(B), tet(G), tet(M), tet(O), tet(Q), and tet(W)], macrolide [erm(B)], methicillin (mecA and mecC), copper (copB, pcoA, pcoD, and tcrB) and zinc (czrC) resistance genes, and three bacterial species (E. coli, Enterococcus spp. and Staphylococcus aureus). The total bacterial population and total abundance of the seven tet genes significantly increased in the three manure types after digestion. Concentration of tet(M) was strongly correlated with that of erm(B) and enterococci. As concentration of tetracyclines declined during anaerobic digestion, that of four tet genes (A, B, Q, and W) and 16S rRNA increased, that of tet(M) decreased, and that of tet(G) and tet(O) did not change. Concentrations of copB and pcoA did not change; while that of pcoD did not change in the PL, it increased in the SM and CM. While the concentration of enterococci remained unchanged in CM, it significantly increased in the PL and SM. Concentrations of tcrB significantly increased in the three manure types. While concentrations of S. aureus significantly increased in the CM and PL, that of SM was not affected. Concentrations of mecC significantly increased in all manure types after digestion; while mecA concentrations did not change in the SM, they significantly increased in CM and PL. While concentration of czrC remained low in the CM, it increased in the PL but declined in the SM. In conclusion, while mesophilic anaerobic digestion of animal manure decreased concentration of tetracyclines, it increased the concentrations of total bacteria, tet genes, E. coli, enterococci and S. aureus and methicillin resistance genes. It did not have any effect on concentrations of heavy metals; concentrations of heavy metal resistance genes either increased or remained unaffected depending on the animal species. This study showed the need for post-digestion treatments of animal manure to remove bacteria, antibiotic resistance genes, heavy metals and their resistance genes.202033391245
3214120.9998Characteristics of the antibiotic resistance genes in the soil of medical waste disposal sites. The inappropriate disposal of medical waste allows bacteria to acquire antibiotic resistance, which results in a threat to public health. Antibiotic resistance gene (ARG) profiles were determined for 45 different soil samples containing medical waste and 15 nearby soil samples as controls. Besides physical and chemical analyses (i.e., dry matter content, pH value, and metal content), the genomes of microorganisms from the soil samples were extracted for high-throughput sequencing. ARG abundances of these samples were obtained by searching the metagenomic sequences against the antibiotic resistance gene database and the copies of ARGs per copy of the 16S rRNA gene at different levels were assessed. The results showed medical waste accumulation significantly enriched the contents of Cu, Cr, Pb, and As in the tested soil samples. Compared to the controls, the samples collected from areas containing medical waste were significantly enriched (p < 0.05, t-test) with ARGs annotated as sulfonamide and multidrug resistance genes, and in particular, the subtypes sul1 and sul2 (sulfonamide resistance genes), and multidrug_transporter (multidrug resistance gene). Moreover, the ARGs of the samples from the polluted areas were more diverse than those of the control samples (p < 0.05, t-test). The comparatively higher abundance and diversity of ARGs in contaminated soil pose a potential risk to human health.202032402966
7199130.9998Minimum influent concentrations of oxytetracycline, streptomycin and spiramycin in selecting antibiotic resistance in biofilm type wastewater treatment systems. It has been demonstrated that antibiotic resistance could be induced and selected under high antibiotic concentrations in biological wastewater treatment systems. However, little is available regarding the minimum concentrations of antibiotics for selecting antibiotic resistance during wastewater treatment. Herein, the minimum influent concentrations of oxytetracycline, streptomycin, and spiramycin in selecting antibiotic resistance in biofilm type wastewater treatment systems were investigated by spiking respective antibiotic into wastewater with an antibiotic dose increasing from 0 to 0.1, 1, 5, 25, 50 mg/L stepwise over a period of 606 days. Significant increase (p < .01) in the total abundance of antibiotic resistance genes was observed for both streptomycin and oxytetracycline at a dose of 0.1 mg/L according to metagenomic sequencing, while the concentration levels leading to significant increases (p < .05) in resistant bacteria ratio were higher: 5 mg/L for streptomycin and 25 mg/L for oxytetracycline. Although resistome abundance increased with the increase of spiramycin dose, neither the corresponding Macrolide-Lincosamide-Streptogramin (MLS) resistance genes nor the resistant bacteria ratio showed perceptible increase. Partial canonical correspondence analysis showed that both bacterial community shift and mobile genetic elements alteration contributed to the enrichment of resistomes under the presence of streptomycin and oxytetracycline. Regarding spiramycin which is mainly targeting on Gram-positive bacteria, the dominance of the intrinsically resisting Gram-negative bacteria in the biofilm microbiota might be responsible for the vague change of MLS resistant determinants under the spiramycin stress. The results demonstrated that it is possible to prevent the development of antibiotic resistance during wastewater treatment by controlling the influent streptomycin and oxytetracyline concentrations below 0.1 mg/L. This work proposed an actionable approach for the management of antibiotic production wastewater.202032325576
3509140.9998Persistence and spatial variation of antibiotic resistance genes and bacterial populations change in reared shrimp in South China. More attention has been paid to the abundance and diversity of antibiotic resistance genes (ARGs) in aquatic environments. However, few studies have investigated the persistence and spatial variation of ARGs in aquatic organisms. This study investigated the occurrence and abundance of ARGs and the bacterial populations in shrimp intestinal tracts during the rearing period in different regions of Guangdong, South China. The results showed that sul1, sul2, qnrD, and floR were the predominant ARGs. Compared with those of juvenile shrimp, the total concentrations of ARGs in the intestinal tract of adult shrimp in three shrimp farms were 2.45-3.92 times higher (p < 0.05), and the bacterial populations in the adult shrimp intestinal tract changed considerably. Redundancy analysis (RDA) showed that the abundance of Proteobacteria, Firmicutes, and Verrucomicrobia in Farms A, B, and C, respectively, were strongly positively correlated with the most abundant and predominant genes (sul1 and qnrD for Farm A; floR and sul2 for Farm B; floR and sul2 for Farm C) in the shrimp intestinal tract. The results of this study indicated that ARGs gained persistence in the developmental stages of the reared shrimp. Different phyla of predominant bacteria were responsible for the increase of ARGs abundance in the shrimp intestinal tract in different regions. This study represents a case study of the persistence and spatial variation of ARGs in aquaculture and can be a reference for the determination of harmful impacts of ARGs on food safety and human health.201829990953
7226150.9998Seasonal variation in antibiotic resistance genes and bacterial phenotypes in swine wastewater during three-chamber anaerobic pond treatment. Antibiotic resistance is a global public health concern. Antibiotic usage in pigs makes swine wastewater (SW) a reservoir for antibiotic resistance genes (ARGs). SW is usually stored and treated in a three-chamber anaerobic pond (3-CAP) in medium and small pig farms in northern China. However, the yet unexplored presence of ARGs in SW during 3-CAP treatment may result in ARGs spreading into the environment if farmers apply SW to farmland as a liquid organic fertilizer. This study investigated the profiles of and changes in ARGs in SW during its treatment in 3-CAP over four seasons and analyzed the correlation between ARGs and bacterial phenotypes, along with the physicochemical parameters of the water. The results revealed that ARG abundance decreased considerably after 3-CAP treatment in April (47%), October (47%), and December (62%) but increased in May (43%) and August (73%). The ARG copies in the influent and other SW samples increased significantly from 10(7) copies/mL in April to 10(9) copies/mL in October and were maintained in December. The increase in ARG abundance was not as rapid as the growth of the bacterial population, resulting in lower relative abundance in October and December. Bacterial communities possessed more sul1 and tetM genes, which were also positively correlated with mobile genetic elements. After the 3-CAP treatment, 16% of antibiotics and 60% of heavy metals were removed, and both had a weak correlation with ARGs. Predicted phenotypes showed that gram-positive (G(+)) and gram-negative (G(-)) bacteria have different capacities for carrying ARGs. G(+) bacteria carry more ARGs than G(-) bacteria. This study revealed the persistence of ARGs in SW after 3-CAP treatment over different seasons. Applying SW in the proper month will mitigate ARG dissemination to the environment.202336208778
7200160.9998Plant Growth, Antibiotic Uptake, and Prevalence of Antibiotic Resistance in an Endophytic System of Pakchoi under Antibiotic Exposure. Antibiotic contamination in agroecosystems may cause serious problems, such as the proliferation of various antibiotic resistant bacteria and the spreading of antibiotic resistance genes (ARGs) in the environment or even to human beings. However, it is unclear whether environmental antibiotics, antibiotic resistant bacteria, and ARGs can directly enter into, or occur in, the endophytic systems of plants exposed to pollutants. In this study, a hydroponic experiment exposing pakchoi (Brassica chinensis L.) to tetracycline, cephalexin, and sulfamethoxazole at 50% minimum inhibitory concentration (MIC) levels and MIC levels, respectively, was conducted to explore plant growth, antibiotic uptake, and the development of antibiotic resistance in endophytic systems. The three antibiotics promoted pakchoi growth at 50% MIC values. Target antibiotics at concentrations ranging from 6.9 to 48.1 µg·kg(-1) were detected in the treated vegetables. Additionally, the rates of antibiotic-resistant endophytic bacteria to total cultivable endophytic bacteria significantly increased as the antibiotics accumulated in the plants. The detection and quantification of ARGs indicated that four types, tetX, bla(CTX-M), and sul1 and sul2, which correspond to tetracycline, cephalexin, and sulfamethoxazole resistance, respectively, were present in the pakchoi endophytic system and increased with the antibiotic concentrations. The results highlight a potential risk of the development and spread of antibiotic resistance in vegetable endophytic systems.201729099753
7119170.9998Assessing the benefits of composting poultry manure in reducing antimicrobial residues, pathogenic bacteria, and antimicrobial resistance genes: a field-scale study. The poultry industry in the European Union produces 13 million tons of manure annually, which represents a major health and environmental challenge. Composting is an environmental-friendly technique for the management of manure, but there are few studies about antibiotic residues and antimicrobial resistances at a field scale. The goal of this study was to determine if the composting of poultry manure at a field scale would result in the reduction of antibiotic residues, pathogenic bacteria, and antibiotic resistance genes (ARGs) in the final fertilizer product. A 10-week composting of poultry manure spiked with enrofloxacin, doxycycline, and ciprofloxacin was performed. The determination of antibiotics residues and 22 selected ARGs was carried out together with the identification of bacteria by metagenomics. In the case of ciprofloxacin and doxycycline, a 90% decrease was observed after composting for 3 weeks. Sixteen ARGs were detected at the beginning of the experiment; 12 of them decreased from week 0 to week 10 (reduction of 73.7-99.99%). The presence of potentially pathogenic bacteria, such as, Campylobacter coli or commensal bacteria such as Escherichia coli decreases along the composting process. In conclusion, 10-week composting of poultry manure promotes the reduction of antibiotic residues and most of the ARGs and pathogenic bacteria.202032399873
7252180.9998Aerobic Composting and Anaerobic Digestion Decrease the Copy Numbers of Antibiotic-Resistant Genes and the Levels of Lactose-Degrading Enterobacteriaceae in Dairy Farms in Hokkaido, Japan. Efficient methods for decreasing the spread of antimicrobial resistance genes (ARGs) and transfer of antimicrobial-resistant bacteria (ARB) from livestock manure to humans are urgently needed. Aerobic composting (AC) or anaerobic digestion (AD) are widely used for manure treatment in Japanese dairy farms. To clarify the effects of AC and AD on antimicrobial resistance, the abundances of antimicrobial (tetracycline and cefazolin)-resistant lactose-degrading Enterobacteriaceae as indicator bacteria, copy numbers of ARGs (tetracycline resistance genes and β-lactamase coding genes), and concentrations of residual antimicrobials in dairy cow manure were determined before and after treatment. The concentration of tetracycline/cefazolin-resistant lactose-degrading Enterobacteriaceae was decreased over 1,000-fold by both AC and AD. ARGs such as tetA, tetB, and bla (TEM) were frequently detected and their copy numbers were significantly reduced by ∼1,000-fold by AD but not by AC. However, several ARG copies remained even after AD treatment. Although concentrations of the majority of residual antimicrobials were decreased by both AC and AD, oxytetracycline level was not decreased after treatment in most cases. In addition, 16S rRNA gene amplicon-based metagenomic analysis revealed that both treatments changed the bacterial community structure. These results suggest that both AC and AD could suppress the transmission of ARB, and AD could reduce ARG copy numbers in dairy cow manure.202134659165
7183190.9998Evaluation of five antibiotic resistance genes in wastewater treatment systems of swine farms by real-time PCR. Antibiotics are widely used in livestock for infection treatment and growth promotion. Wastes from animal husbandry are a potential environmental source of antibiotic-insensitive pathogens, and the removal efficiency of the resistance genotypes in current wastewater treatment plants (WWTPs) is unknown. In this study, quantitative PCR was used for evaluating antibiotic resistance genes in wastewater treatment processes. Six wastewater treatment plants in different swine farms were included in this study, and five antibiotic resistance genes (ARGs) were tested for each treatment procedure. All of the tested ARGs including tetA, tetW, sulI, sulII, and blaTEM genes were detected in six swine farms with considerable amounts. The results showed that antibiotic resistance is prevalent in livestock farming. The ARG levels were varied by wastewater treatment procedure, frequently with the highest level at anaerobic treatment tank and lowest in the activated sludge unit and the effluents. After normalizing the ARG levels to 16S rRNA gene copies, the results showed that ARGs in WWTP units fluctuated partly with the quantity of bacteria. Regardless of its importance in biodegradation, the anaerobic procedure may facilitate bacterial growth thus increasing the sustainability of the antibiotic resistance genotypes. After comparing the copy numbers in influx and efflux samples, the mean removal efficiency of ARGs ranged between 33.30 and 97.56%. The results suggested that treatments in the WWTP could partially reduce the spread of antibiotic-resistant bacteria, and additional procedures such as sedimentation may not critically affect the removal efficiency.201425064719