Distribution of antibiotic resistance genes in Bosten Lake, Xinjiang, China. - Related Documents




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525901.0000Distribution of antibiotic resistance genes in Bosten Lake, Xinjiang, China. The occurrence of antibiotic resistance genes (ARGs) and resistant bacteria was quantified in 17 water samples collected across Bosten Lake, Xinjiang, China. The heterotrophic plate count method was used to detect the levels of sulfonamide- and tetracycline-resistant bacteria, which have mean concentrations of 2.50×10(5) and 4.63×10(3) CFU/mL, respectively. The resistance genes of sulfonamide (sul1, sul2) and tetracycline (tetM, tetO and tetW) were detected, and results showed that all other ARGs except the tetO gene were obtained from all samples. Four of the obtained ARGs were further quantified, and results showed that the sulfonamide resistance genes were prevalent. The relative abundance was in the range of 2.81×10(-5) to 3.33×10(-3) for the sul1/16s-rRNA and 1.04×10(-5) to 3.80×10(-3) for the sul2/16s-rRNA. For the tet genes, the relative concentrations of tetM/16s-rRNA and tetW16s-rRNA ranged from 1.18×10(-5) to 2.46×10(-4) and 1.58×10(-6) to 4.19×10(-4), respectively. The concentration divergence among ARGs may be related to the different characteristics of each gene. This study validated that Bosten Lake was affected by ARGs and resistant bacteria, thus turning the lake into an important reservoir for the transfer of ARGs and resistant bacteria.201425225942
525710.9998Removal of fecal indicator bacteria and antibiotic resistant genes in constructed wetlands. Wastewater discharge evidently increased bacterial diversity in the receiving waterbodies. The objective of this study was to evaluate the effectiveness of a constructed wetland in reducing fecal indicator bacteria (FIB) and antibiotic resistant genes (ARGs). We determined the prevalence and attenuation of fecal indicator bacteria including Escherichia coli and enterococci, along with ARGs, and human-associated Bacteroidales (HF183) markers by quantitative polymerase chain reaction (qPCR) method. Three types of water samples (inlet, intermediate, and outlet) from a constructed wetland were collected once a month from May to December in 2013. The overall reduction of E. coli was 50.0% based on culture method. According to the qPCR result, the overall removal rate of E. coli was only 6.7%. Enterococci were found in 62.5% of the wetland samples. HF183 genetic marker was detected in all final effluent samples with concentration ranging from 1.8 to 4.22 log(10) gene copies (GC)/100 ml. Of the ARGs tested, erythromycin resistance genes (ermF) were detected in 79.2% of the wetland samples. The class 1 integrase (intI1) was detected in all water samples with concentration ranging from 0.83 to 5.54 log(10) GC/100 ml. The overall removal rates of enterococci, HF183, intI1, and ermF were 84.0%, 66.6%, 67.2%, and 13.1%, respectively.201930758793
526020.9998Occurrence and Abundance of Antibiotic Resistance Genes in Chinese Traditional Pickles. With the widespread application and even misuse of antibiotics, antibiotic resistance genes (ARGs) are extensively present in various environments, from natural environment to fermented foods, posing emerging threat to public and environmental health. The real-time fluorescence quantitative PCR (qPCR) technique is commonly used to detect ARGs of environmental samples such as soil or water. In this study, eight types of pickles were collected from four regions of China and the existence of 13 resistance genes was assessed by qPCR. The results showed that a total of 11 resistance genes were detected in pickles, the blaTEM gene was detected in all samples, and the neo and cat genes were absent. The abundance of resistance genes varied, aada1 (1.09 × 10(5) to 5.94 × 10(6) copies/g), blaTEM (1.48 × 10(5) to 2.2 × 10(6) copies/g), ermc (1.01 × 10(5) to 5.35 × 10(5) copies/g), hyg (1.35 × 10(5) to 1.93 × 10(6) copies/g), aadd (4.46 × 10(5) to 1.60 × 10(6) copies/g), nat1 (1.04 × 10(5) to 5.04 × 10(5) copies/g), nptII (2.17 × 10(4) to 1.69 × 10(5) copies/g), sul1 (2.01 × 10(5) to 4.60 × 10(5) copies/g), tetl (1.23 × 10(5) to 6.18 × 10(5) copies/g), shble (1.68 × 10(4) copies/g), and stra (4.8 × 10(4) to 1.9 × 10(5)copies/g). We also discussed the specificity and sensitivity assessment of qPCR applied to ARGs analysis in pickles, verifying the feasibility and validity of the method. Bacteria were isolated and purified from pickles as well and their antimicrobial resistance was studied. This study is of great significance for the risk assessment of resistance genes in pickles. Effective and preventive solutions were proposed to reduce the spread of resistance genes and protect public dietary health.202540230011
525830.9998Occurrence of seventeen veterinary antibiotics and resistant bacterias in manure-fertilized vegetable farm soil in four provinces of China. This study focused on the occurrence of seventeen veterinary antibiotics and six resistant bacterias in soils from the vegetable farms fertilized with animal manure in China. Seventeen veterinary antibiotics, including sulfonamides, quinolones, tetracyclines, macrolides and amphenicols, were detected by high performance liquid chromatography/tandem mass spectrometer in all the 53 soil samples collected in four provinces during August 2016. The concentrations of target antibiotics in the soil samples ranged from not detectable to 415.00 μg/kg dry weight with the mean residual levels of the five classes followed order: tetracyclines (82.75 μg/kg) > quinolones (12.78 μg/kg) > macrolides (12.24 μg/kg) > sulfonamides (2.61 μg/kg) > amphenicols (0.06 μg/kg). Moreover, the highest antibiotic levels were found mainly in soil from organic vegetable farms. Risk assessment by using the methods of risk quotient, suggested that oxytetracycline, chlortetracycline, enrofloxacin and ciprofloxacin could pose severe ecological risk in sampled soils. Resistant strains were isolated in 30 samples, with Escherichia coli and Klebsiella pneumonia found the dominant bacterial hosts with resistance genes. Antibiotic resistance genes, including tetA, tetB, qnrS, oqxA, sul1, sul2, ermA and floR, were detected in the strains resistant to: tetracyclines, quinolones, sulfonamides, macrolides and amphenicols resistance, respectively. Overall, there was a correlation between the results of antibiotic risk assessment with the detection of resistance genes from isolated strains in the soils.201930317094
777240.9998Metagenomic community composition and resistome analysis in a full-scale cold climate wastewater treatment plant. BACKGROUND: Wastewater treatment plants are an essential part of maintaining the health and safety of the general public. However, they are also an anthropogenic source of antibiotic resistance genes. In this study, we characterized the resistome, the distribution of classes 1-3 integron-integrase genes (intI1, intI2, and intI3) as mobile genetic element biomarkers, and the bacterial and phage community compositions in the North End Sewage Treatment Plant in Winnipeg, Manitoba. Samples were collected from raw sewage, returned activated sludge, final effluent, and dewatered sludge. A total of 28 bacterial and viral metagenomes were sequenced over two seasons, fall and winter. Integron-integrase genes, the 16S rRNA gene, and the coliform beta-glucuronidase gene were also quantified during this time period. RESULTS: Bacterial classes observed above 1% relative abundance in all treatments were Actinobacteria (39.24% ± 0.25%), Beta-proteobacteria (23.99% ± 0.16%), Gamma-proteobacteria (11.06% ± 0.09%), and Alpha-proteobacteria (9.18 ± 0.04%). Families within the Caudovirales order: Siphoviridae (48.69% ± 0.10%), Podoviridae (23.99% ± 0.07%), and Myoviridae (19.94% ± 0.09%) were the dominant phage observed throughout the NESTP. The most abundant bacterial genera (in terms of average percent relative abundance) in influent, returned activated sludge, final effluent, and sludge, respectively, includes Mycobacterium (37.4%, 18.3%, 46.1%, and 7.7%), Acidovorax (8.9%, 10.8%, 5.4%, and 1.3%), and Polaromonas (2.5%, 3.3%, 1.4%, and 0.4%). The most abundant class of antibiotic resistance in bacterial samples was tetracycline resistance (17.86% ± 0.03%) followed by peptide antibiotics (14.24% ± 0.03%), and macrolides (10.63% ± 0.02%). Similarly, the phage samples contained a higher prevalence of macrolide (30.12% ± 0.30%), peptide antibiotic (10.78% ± 0.13%), and tetracycline (8.69% ± 0.11%) resistance. In addition, intI1 was the most abundant integron-integrase gene throughout treatment (1.14 × 10(4) gene copies/mL) followed by intI3 (4.97 × 10(3) gene copies/mL) while intI2 abundance remained low (6.4 × 10(1) gene copies/mL). CONCLUSIONS: Wastewater treatment successfully reduced the abundance of bacteria, DNA phage and antibiotic resistance genes although many antibiotic resistance genes remained in effluent and biosolids. The presence of integron-integrase genes throughout treatment and in effluent suggests that antibiotic resistance genes could be actively disseminating resistance between both environmental and pathogenic bacteria.202235033203
777450.9998Antibiotic-resistant genes and antibiotic-resistant bacteria in the effluent of urban residential areas, hospitals, and a municipal wastewater treatment plant system. In this study, we determined the abundance of 8 antibiotics (3 tetracyclines, 4 sulfonamides, and 1 trimethoprim), 12 antibiotic-resistant genes (10 tet, 2 sul), 4 antibiotic-resistant bacteria (tetracycline, sulfamethoxazole, and combined resistance), and class 1 integron integrase gene (intI1) in the effluent of residential areas, hospitals, and municipal wastewater treatment plant (WWTP) systems. The concentrations of total/individual targets (antibiotics, genes, and bacteria) varied remarkably among different samples, but the hospital samples generally had a lower abundance than the residential area samples. The WWTP demonstrated removal efficiencies of 50.8% tetracyclines, 66.8% sulfonamides, 0.5 logs to 2.5 logs tet genes, and less than 1 log of sul and intI1 genes, as well as 0.5 log to 1 log removal for target bacteria. Except for the total tetracycline concentration and the proportion of tetracycline-resistant bacteria (R (2) = 0.330, P < 0.05), there was no significant correlation between antibiotics and the corresponding resistant bacteria (P > 0.05). In contrast, various relationships were identified between antibiotics and antibiotic resistance genes (P < 0.05). Tet (A) and tet (B) displayed noticeable relationships with both tetracycline and combined antibiotic-resistant bacteria (P < 0.01).201525323405
525660.9997Characterization of antibiotic resistance genes and bacteria in a municipal water resource recovery facility. Municipal water resource recovery facilities (WRRFs) are important sources of antibiotic-resistant bacteria and genes (ARB and ARGs). In this study, antibiotic-resistant total heterotrophic bacteria (THB(R) ) counts (CFU/ml) cultivated from influent, effluent of activated sludge process, and outflow of disinfection unit of an urban WRRF were investigated for the presence of 16, 32, 64, and 128 μg/ml of nine antibiotics. The isolates of Pseudomonas spp., Acinetobacter spp., and Escherichia coli obtained from effluent of activated sludge process were subjected for molecular identification by detecting the 16S rRNA gene sequences. Additionally, using the polymerase chain reaction method (PCR), the isolates were investigated for the presence of bla(SHV) , bla(TEM) , bla(CTX-M) , bla(VIM) , sul1, and qnrS genes. According to the results, the abundance of THB(R) counts was not significantly reduced by the biological treatment except for cefixime and sulfamethoxazole; it also increased for some antibiotics after disinfection unit. The average removal efficiency of THB(R) resistant to ciprofloxacin, sulfamethoxazole, and ceftazidime were 7.9 ± 1.7%, 41.8 ± 2.1%, and 14.4 ± 6.2%, respectively. Also, all the tested isolates were resistant to at least four antibiotics. For all antibiotics, the resistance ratio (THB(R) /THB) significantly increased in the effluent and after chlorination unit. Among 12 resistant isolates, bla(TEM) and sul1 genes were the most frequently detected ones involved in 92% and 83% of the isolates, respectively. Both bla(TEM) and sul1 genes were found in 100% of E. coli, and 83% and 67% of Pseudomonas spp. isolates, respectively. Further efforts are necessary to limit the transmission of ARB and ARGs from WRRFs into the environment and prevent human health threats. PRACTITIONER POINTS: The ratio of resistance significantly increased after biological treatment. Up to 40% of heterotrophic bacteria in the effluent was antibiotic resistant. bla(TEM) and sul1 genes were more prevalent (92%) in all isolates of bacteria. Both bla(TEM) and sul1 genes were found in 100% of E. coli isolates. Pseudomonas spp. holds bla(TEM) and sul1 genes in 83% and 67% of isolates, respectively.202235765862
777370.9997Correlation of tetracycline and sulfonamide antibiotics with corresponding resistance genes and resistant bacteria in a conventional municipal wastewater treatment plant. Antibiotics and corresponding resistance genes and resistant bacteria have been considered as emerging pollutants worldwide. Wastewater treatment plants (WWTPs) are potential reservoirs contributing to the evolution and spread of antibiotic resistance. In this study, total concentrations of tetracycline and sulfonamide antibiotics in final effluent were detected at 652.6 and 261.1ng/L, respectively, and in treated sludge, concentrations were at 1150.0 and 76.0μg/kg dry weight (dw), respectively. The quantities of antibiotic resistance genes and antibiotic resistant bacteria in final effluent were quantified in the range of 9.12×10(5)-1.05×10(6) gene abundances /100mL (genomic copies/100mL) and 1.05×10(1)-3.09×10(3)CFU/mL, respectively. In treated sludge, they were quantified at concentrations of 1.00×10(8)-1.78×10(9) gene abandances/100mL and 7.08×10(6)-1.91×10(8)CFU/100mL, respectively. Significant reductions (2-3 logs, p<0.05) of antibiotic resistance genes and antibiotic resistant bacteria were observed between raw influent and final effluent. The gene abundances of tetO and tetW normalized to that of 16S rRNA genes indicated an apparent decrease as compared to sulI genes, which remained stable along each treatment stage. Significant correlations (R(2)=0.75-0.83, p<0.05) between numbers of resistant bacteria and antibiotic concentrations were observed in raw influent and final effluent. No significance (R(2)=0.15, p>0.05) was found between tet genes (tetO and tetW) with concentration of tetracyclines identified in wastewater, while a significant correlation (R(2)=0.97, p<0.05) was observed for sulI gene and total concentration of sulfonamides. Correlations of the quantities of antibiotic resistance genes and antibiotic resistant bacteria with corresponding concentrations of antibiotics in sludge samples were found to be considerably weak (R(2)=0.003-0.07).201222369865
308880.9997Occurrence of sulfonamide and tetracycline-resistant bacteria and resistance genes in aquaculture environment. The occurrence of sulfonamide and tetracycline resistance and their pollution profile in the aquaculture environment of Tianjin, northern China, were investigated. The presence of antibiotic-resistant bacteria was identified and the corresponding antibiotic resistance genes (ARGs) were quantified at 6 aquaculture farms in Tianjin. Sulfonamide-resistance genes were prevalent and their concentrations were the highest detected (3.0 × 10(-5) to 3.3 × 10(-4) for sul1/16S rDNA, 2.0 × 10(-4) to 1.8 × 10(-3) for sul2/16S rDNA) among the various ARGs, most likely because the use of sulfonamides is more prevalent than tetracyclines in this area. Bacillus was the most dominant bacterial genus in both sulfamethoxazole resistant bacteria (63.27% of the total resistant bacteria) and tetracycline-resistant bacteria (57.14% of the total resistant bacteria). At least two of those genes (tetM, tetO, tetT, tetW, sul1 and sul2) were detected in the isolates of Bacillus cereus, Bacillus subtilis, Bacillus megaterium and Acinetobacter lwofii, and all of the above genes were detected in B. cereus, suggesting the occurrence of multi-resistance in the studied area. The genetic transfer of sul1 between intestinal bacteria (e.g., Enterococcus spp.) and indigenous bacteria (e.g., Bacillus spp.) was implied by phylogenetic analysis. Several strains of resistant opportunistic pathogens (e.g., Acinetobacter spp.) were found in indigenous bacteria, which increase the risk of ARGs to public health. Overall, this is the first study to comprehensively investigate the antibiotic resistance profile by analyzing the species of antibiotic-resistant bacteria and adopting qualitative and quantitative methods to investigate ARGs at a typical aquaculture area in northern China.201222377146
525290.9997Prevalence of sulfonamide resistance genes in bacterial isolates from manured agricultural soils and pig slurry in the United Kingdom. The prevalences of three sulfonamide resistance genes, sul1, sul2, and sul3 and sulfachloropyridazine (SCP) resistance were determined in bacteria isolated from manured agricultural clay soils and slurry samples in the United Kingdom over a 2-year period. Slurry from tylosin-fed pigs amended with SCP and oxytetracycline was used for manuring. Isolates positive for sul genes were further screened for the presence of class 1 and 2 integrons. Phenotypic resistance to SCP was significantly higher in isolates from pig slurry and postapplication soil than in those from preapplication soil. Of 531 isolates, 23% carried sul1, 18% sul2, and 9% sul3 only. Two percent of isolates contained all three sul genes. Class 1 and class 2 integrons were identified in 5% and 11.7%, respectively, of sul-positive isolates. In previous reports, sul1 was linked to class 1 integrons, but in this study only 8% of sul1-positive isolates carried the intI1 gene. Sulfonamide-resistant pathogens, including Shigella flexneri, Aerococcus spp., and Acinetobacter baumannii, were identified in slurry-amended soil and soil leachate, suggesting a potential environmental reservoir. Sulfonamide resistance in Psychrobacter, Enterococcus, and Bacillus spp. is reported for the first time, and this study also provides the first description of the genotypes sul1, sul2, and sul3 outside the Enterobacteriaceae and in the soil environment.200919064898
3515100.9997Impact of reclaimed water irrigation on antibiotic resistance in public parks, Beijing, China. The abundance and distribution of antibiotics and antibiotic resistance genes (ARGs) in soils from six parks using reclaimed water in Beijing, China, were characterized. Three classes of commonly used antibiotics (tetracycles, quinolones, and sulfonamides) were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The highest concentrations of tetracyclines and quinolones were 145.2 μg kg(-1) and 79.2 μg kg(-1), respectively. Detected tetG, tetW, sulI, and sulII genes were quantified by quantitative PCR. ARGs exhibited various abundances for different park soils. The integrase gene (intI1) as an indicator of horizontal gene transfer potential was also detected in high abundance, and had significant positive correlation with tetG, sulI, and sulII genes, suggesting that intI1 may be involved in ARGs dissemination. Both sulII and intI1 clones had high homology with some classes of pathogenic bacteria, such as Klebsiella oxytoca, Acinetobacter baumannii, Shigella flexneri, which could trigger potential public health concern.201424071635
3089110.9997Distribution characteristics of antibiotic resistance bacteria and related genes in urban recreational lakes replenished by different supplementary water source. The distribution characteristics of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in urban recreational water from different water-supply sources might be different. In this study, water samples were collected to detect the antibiotic resistance of heterotrophic bacteria to five antibiotics, and the content, phenotype, gene type and species distribution of resistant bacteria were analyzed. The results showed that the changes of bacteria resistance rate in two lakes to five kinds of antibiotics were synchronous with time, and it would reach its maximum in autumn. The detection of ARGs and int I in 80 resistance strains showed that the detection rate of tetG, tetA and int I was high. Here, 51.25% of the bacteria were doubly resistant to AMP-CTX. The 80 isolate strains were of nine genera and 19 species, among which Bacillus cereus, Escherichia coli, Aeromonas veronii, Aeromonas caviae and Raoultella ornithinolytica were the common ARB species in two lakes. Correlation analysis showed that the water temperature was significantly correlated with the content of ARB in sulfamethoxazole (SMZ) and cefotaxime (CTX) (p < 0.05), and the total phosphorus (TP) in FQ lake was significantly correlated with the content of AMP-resistant bacteria (p < 0.05), while there were no other correlations between the changes of other water quality indexes and the content of ARB (p > 0.05).202235228362
3087120.9997Diversity and abundance of antibiotic resistance of bacteria during the seedling period in marine fish cage-culture areas of Hainan, China. Antibiotic resistance has become an important focus of research in the aquaculture environment. However, few studies have evaluated antibiotic resistance during the seedling period in marine fish cage-culture areas. In this study, culture-dependent methods and quantitative polymerase chain reaction were used to identify and detect cultivable heterotrophic antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs), respectively, during the seedling period in a marine fish cage-culture areas of Hainan, China. Bacterial resistance to amoxicillin, erythromycin, and gentamicin was generally high (average on 27.67%, 23.61% and 37.32%, respectively), whereas resistance to furazolidone and nitrofurantoin was generally low (average on 0.14% and 7.425%). Alteromonas (32.72%) and Vibrio (24.77%) were the dominant genus of ARB. Most ARB were opportunistic pathogens, belonging to the phylum Proteobacteria (96.02%). The abundance of sul family genes was higher than that of tet family genes. Overall, the abundance of ARGs and the resistance rates in HW was highest.201930955742
5271130.9997Occurrence of Antibiotic-Resistant Bacteria and Genes in Two Drinking Water Treatment and Distribution Systems in the North-West Province of South Africa. With the increasing spread of antimicrobial resistance, there is growing attention to the contribution made by drinking water systems. The potential health impact of two drinking water treatment and distribution systems (A and B) in the North-West Province of South Africa was determined by investigating the water quality and occurrence of antimicrobial-resistant heterotrophic bacteria and genes in the raw and treated water over four seasons. Most of the physicochemical parameters except for electrical conductivity were within permissible limits. Coliform bacteria reduced from raw to potable water except for counts higher than the threshold recorded in Summer and Winter. A total of 203 heterotrophic bacterial isolates were recovered on chromogenic R2A medium and subjected to susceptibility testing to twelve antibiotics. Most of the isolates were resistant to β-lactam antibiotics and Trimethoprim, whereas they were susceptible to Ciprofloxacin, Erythromycin, and Neomycin. The proportions of Cephalothin and Kanamycin-resistant isolates were significantly higher (p < 0.05) after treatment for site A, compared to significantly lower β-lactam, Oxytetracycline, and Trimethoprim-resistant isolates for B. Over 50% of isolates were of high risk, indicating their origin from high antibiotic-use sources. Seventy-one (35%) isolates were multidrug-resistant, out of which the majority (53.5%, n = 38) possessed the strA gene, followed by strB 21 (29.6%), dfrB 13 (18.3%), aadA 11 (15.5%), bla(CTX-M) 5 (7.0%), and tetA 3 (4.2%). The 16S rRNA gene sequences of the isolates revealed strains belonging to eight bacterial families, some of which are clinically important.202033126462
5255140.9997Occurrence and removal of antibiotics, antibiotic resistance genes, and bacterial communities in hospital wastewater. Hospital wastewater contains a variety of human antibiotics and pathogens, which makes the treatment of hospital wastewater essential. However, there is a lack of research on these pollutants at hospital wastewater treatment plants. In this study, the characteristics and removal of antibiotics and antibiotic resistance genes (ARGs) in the independent treatment processes of hospitals of different scales (primary hospital, H1; secondary hospital, H2; and tertiary hospital, H3) were investigated. The occurrence of antibiotics and ARGs in wastewater from three hospitals varied greatly. The first-generation cephalosporin cefradine was detected at a concentration of 2.38 μg/L in untreated wastewater from H1, while the fourth-generation cephalosporin cefepime had the highest concentration, 540.39 μg/L, at H3. Ofloxacin was detected at a frequency of 100% and had removal efficiencies of 44.2%, 51.5%, and 81.6% at H1, H2, and H3, respectively. The highest relative abundances of the β-lactam resistance gene bla(GES-1) (1.77×10(-3) copies/16S rRNA), the quinolone resistance gene qnrA (8.81×10(-6) copies/16S rRNA), and the integron intI1 (1.86×10(-4) copies/16S rRNA) were detected in the treated wastewater. The concentrations of several ARGs were increased in the treated wastewater (e.g. bla(OXA-1), bla(OXA-10), and bla(TEM-1)). Several pathogenic or opportunistic bacteria (e.g. Acinetobacter, Klebsiella, Aeromonas, and Pseudomonas) were observed at high relative abundances in the treated wastewater. These results suggested the co-occurrence of antibiotics, ARGs, and antibiotic-resistant pathogens in hospital wastewater, and these factors may spread into the receiving aquatic environment.202134089156
5253150.9997Effects of Cage Farming on Antimicrobial and Heavy Metal Resistance of Escherichia coli, Enterococcus faecium, and Lactococcus garvieae. OBJECTIVE: To characterize antibiotic resistance genes (ARGs) and heavy metal resistance genes (HMRGs) of Escherichia coli and Enterococcus faecium isolated from the sediment and Lactococcus garvieae isolated from fish. MATERIALS AND METHODS: The isolated bacteria were identified by sequencing 16S rRNA genes. After identification of the bacteria, tetracycline (tetA, tetB, tetD), erythromycin (ereA, ereB), sulfonamides (sulI, sulII), trimethoprim (dhfrA1), β-lactam (bla(TEM), bla(CTX), ampC), florfenicol (floR), and class 1 integron (Int1) resistance gene were then determined. The presence of HMRGs, including copper (copA), mercury (mer), cadmium, zinc, cobalt (czc), and nickel, cobalt cadmium (ncc), was also analyzed by PCR. All strains were checked for the presence of ARGs and/or HMRGs on the plasmid. RESULTS: The frequency of the β-lactam resistance gene was highest and ranged from 49.7% to 62.3%, followed by sulfonamides, tetracyclines, phenicols, and macrolide resistance genes. The cage culture fish farming practice showed significant effects on ARG frequency of bacteria isolated from the sediment, whereas it had no effect on the frequency of HMRGs. The most prevalent HMRG was determined as mercury-resistant mer gene in all bacteria. All four of the HMRGs were located on plasmids with frequency ranging from 1.20% to 32.53%. The presence of ARGs on plasmids ranged between 2.2% (Dhfr1) and 75% (AmpC, blactx, tetB), and plasmids did not contain tetD and ereB genes. CONCLUSION: The results of this study indicate that fish farming can significantly influence the antimicrobial resistance properties of bacteria isolated from sediment samples.201829733265
5262160.9997High abundances of class 1 integrase and sulfonamide resistance genes, and characterisation of class 1 integron gene cassettes in four urban wetlands in Nigeria. There is little information about environmental contamination with antibiotic resistance genes (ARG) in Sub-Saharan Africa, home to about 1 billion people. In this study we measured the abundance of three genes (sul1, sul2, and intI1) used as indicators of environmental contamination with ARGs in the sediments of four urban wetlands in southwestern Nigeria by qPCR. In addition, we characterised the variable regions of class 1 integrons in sulfamethoxazole/trimethoprim (SMX/TRI)-resistant bacteria isolated from the wetlands by PCR and DNA sequencing. The indicator ARGs were present in all wetlands with mean absolute copy numbers/gram of sediment ranging between 4.7x106 and 1.2x108 for sul1, 1.1x107 and 1x108 for sul2, and 5.3x105 and 1.9x107 for intI1. The relative abundances (ARG/16S rRNA copy number) ranged from about 10-3 to 10-1. These levels of ARG contamination were similar to those previously reported for polluted environments in other parts of the world. The integrase genes intI1 and intI2 were detected in 72% and 11.4% SMX/TRI-resistant isolates, respectively. Five different cassette array types (dfrA7; aadA2; aadA1|dfrA1; acc(6')lb-cr|arr3|dfrA27; arr3|acc(6')lb-cr|dfrA27) were detected among 34 (59.6%) intI1-positive isolates. No gene cassettes were found in the nine intI2-positive isolates. These results show that African urban ecosystems impacted by anthropogenic activities are reservoirs of bacteria harbouring transferable ARG.201830496274
5261170.9997Prevalence of antibiotic resistance genes from effluent of coastal aquaculture, South Korea. The wide use of antibiotics in aquaculture for prophylactic and therapeutic purposes can potentially lead to the prevalence of antibiotic resistance genes (ARGs). This study reports for the first time the profile of ARGs from effluents of coastal aquaculture located in South Jeolla province and Jeju Island, South Korea. Using quantitative PCR (qPCR), twenty-two ARGs encoding tetracycline resistance (tetA, tetB, tetD, tetE, tetG, tetH, tetM, tetQ, tetX, tetZ, tetBP), sulfonamide resistance (sul1, sul2), quinolone resistance (qnrD, qnrS, aac(6')-Ib-cr), β-lactams resistance (bla(TEM), bla(CTX), bla(SHV)), macrolide resistance (ermC), florfenicol resistance (floR) and multidrug resistance (oqxA) and a class 1 integrons-integrase gene (intI1) were quantified. In addition, Illumina Miseq sequencing was applied to investigate microbial community differences across fish farm effluents. Results from qPCR showed that the total number of detected ARGs ranged from 4.24 × 10(-3) to 1.46 × 10(-2) copies/16S rRNA gene. Among them, tetB and tetD were predominant, accounting for 74.8%-98.0% of the total ARGs. Furthermore, intI1 gene showed positive correlation with tetB, tetD, tetE, tetH, tetX, tetZ tetQ and sul1. Microbial community analysis revealed potential host bacteria for ARGs and intI1. Two genera, Vibrio and Marinomonas belonging to Gammaproteobacteria, showed significant correlation with tetB and tetD, the most dominant ARGs in all samples. Also, operational taxonomic units (OTUs)-based network analysis revealed that ten OTUs, classified into the phyla Proteobacteria, Cyanobacteria/Chloroplast, Bacteroidetes, Verrucomicrobia and an unclassified phylum, were potential hosts of tetracycline resistance genes (i.e., tetA, tetG, tetH, tetM, tetQ and tetZ). Further systematic monitoring of ARGs is warranted for risk assessment and management of antibacterial resistance from fish farm effluents.201829031406
5249180.9996Characteristics of heterotrophic endophytic bacteria in four kinds of edible raw vegetables: species distribution, antibiotic resistance, and related genes. This study aimed to explore antibiotic resistance characteristics and species of heterotrophic endophytic bacteria (HEB) in four kinds of edible raw vegetables, including radishes, lettuces, onions, and tomatoes. A total of 144 HEB were isolated and tested for resistance to sulfamethoxazole (SMZ), tetracycline (TET), cefotaxime (CTX), and ciprofloxacin (CIP), and their species were identified by 16S rRNA gene sequencing. Antibiotic resistance genes (ARGs) and class I integron in antibiotic-resistant isolates were analyzed by polymerase chain reaction. The results showed radishes had the highest, while tomatoes had the lowest concentration of antibiotic-resistant HEB. SMZ and CTX were predominant antibiotic-resistant phenotypes in HEB. The multi-resistant phenotypes, the combinations SMZ-TET-CTX and SMZ-TET-CIP, accounted for 9.34% of all antibiotic-resistant phenotypes, mainly in radishes and lettuces. Bacillus, Pseudomonas, Staphylococcus, and Stenotrophomonas showed resistance to two antibiotics and existed in more than one kind of vegetable, and were the main carriers of sul1, sul2, blaTEM, and intI1 genes. Therefore, these four genera were considered potential hosts of ARGs in edible raw vegetables. The study provides an early warning regarding health risks associated with ingesting antibiotic-resistant bacteria through raw vegetable consumption.202439611313
2827190.9996Characterization of microbiota composition and presence of selected antibiotic resistance genes in carriage water of ornamental fish. International trade with ornamental fish is gradually recognized as an important source of a wide range of different antibiotic resistant bacteria. In this study we therefore characterized the prevalence of selected antibiotic resistance genes in the microbiota found in the carriage water of ornamental fish originating from 3 different continents. Real-time PCR quantification showed that the sul1 gene was present in 11 out of 100 bacteria. tet(A) was present in 6 out of 100 bacteria and strA, tet(G), sul2 and aadA were present in 1-2 copies per 100 bacteria. Class I integrons were quite common in carriage water microbiota, however, pyrosequencing showed that only 12 different antibiotic gene cassettes were present in class I integrons. The microbiota characterized by pyrosequencing of the V3/V4 variable region of 16S rRNA genes consisted of Proteobacteria (48%), Bacteroidetes (29.5%), Firmicutes (17.8%), Actinobacteria (2.1%) and Fusobacteria (1.6%). Correlation analysis between antibiotic resistance gene prevalence and microbiota composition verified by bacterial culture showed that major reservoirs of sul1 sul2, tet(A), tet(B) tet(G), cat, cml, bla, strA, aacA, aph and aadA could be found among Alpha-, Beta- and Gammaproteobacteria with representatives of Enterobacteriaceae, Pseudomonadaceae, Rhizobiaceae and Comamonadaceae being those most positively associated with the tested antibiotic resistance genes.201425084116