Analysis of antibiotic resistance in bacteria isolated from the surface microlayer and underlying water of an estuarine environment. - Related Documents




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289801.0000Analysis of antibiotic resistance in bacteria isolated from the surface microlayer and underlying water of an estuarine environment. We compared the prevalence of cultivable antibiotic-resistant bacteria and resistance genes in the surface microlayer (SML) and underlying waters (UW) of an estuary. Prevalence of resistant bacteria was determined in antibiotic-supplemented agar. Bacterial isolates from the UW (n=91) and SML (n=80), selected in media without antibiotic, were characterized concerning susceptibility against nine antibiotics. The presence of genes bla(TEM), bla(OXA-B), bla(SHV), bla(IMP), tet(A), tet(B), tet(E), tet(M), cat, sul1, sul2, sul3, aadA, IntI1, IntI2, and IntI3 was assessed by PCR. The variable regions of integrons were sequenced. Ampicillin- and streptomycin-resistant bacteria were significantly more prevalent in SML. Resistance levels among the bacterial collections were generally low, preventing detection of significant differences between SML and UW. The tet(E) gene was detected in two Aeromonas isolates and tet(M) was detected in a Pseudomonas isolate. Gene sul1 was amplified from three Aeromonas isolates. Prevalence of intI genes was 2.11%. Cassette arrays contained genes encoding resistance to aminoglycosides and chloramphenicol. A higher prevalence of antibiotic-resistant bacteria in the SML, although only detectable when bacteria were selected in antibiotic-supplemented agar, suggests that SML conditions select for antibiotic resistance. Results also showed that antibiotic resistance was uncommon among estuarine bacteria and the resistance mechanisms are probably predominantly intrinsic.201323067198
285110.9997Detection of antibiotic resistance and tetracycline resistance genes in Enterobacteriaceae isolated from the Pearl rivers in South China. This study investigated antibiotic resistance profiles and tetracycline resistance genes in Enterobacteriaceae family isolates from the Pearl rivers. The Enterobacteriaceae isolates were tested for susceptibility to seven antibiotics ampicillin, chloramphenicol, ciprofloxacin, levofloxacin, sulphamethoxazole/trimethoprim, tetracycline and trimethoprim. In Liuxi reservoir, with an exception to ampicillin resistant strains (11%) no other antibiotic resistance bacterial strains were detected. However, multiple drug resistance in bacterial isolates from the other sites of Pearl rivers was observed which is possibly due to sewage discharge and input from other anthropogenic sources along the rivers. Four tetracycline resistance genes tet A, tet B, tet C and tet D were detected in the isolates from the rivers. The genes tet A and tet B were widely detected with the detection frequencies of 43% and 40% respectively. Ciprofloxacin and levofloxacin resistant enteric bacteria were also isolated from the pig and duck manures which suggest a wider distribution of human specific drugs in the environment. This investigation provided a baseline data on antibiotic resistance profiles and tetracycline resistance genes in the Pearl rivers delta.201020356660
527020.9997Environmental antibiotic stress and high-risk resistance genes in bacterial communities of the Gomti and Ganga Rivers, India. BACKGROUND: The river ecosystems provide habitats and source of water for a number of species including humans. The uncontrolled accumulation of pollutants in the aquatic environment enhances the development of antibiotic-resistant bacteria and genes. METHODS: Water samples were collected seasonally from different sites of Gomti and Ganga River. Bacteria were isolated by plating on nutrient agar supplemented with individual antibiotics (100 µg/ml) to select the resistant strains. These isolates were subsequently tested for cross-resistance to other antibiotics using the disc diffusion method. PCR was performed to detect selected ARGs. RESULTS: The enumeration of microbial population of Gomti River, the tetracycline-resistant bacteria comprised 38% of the bacterial population during spring and chloramphenicol resistance during autumn was a mere 11.9%. Nevertheless, erythromycin resistance was widespread amongst Ganga river bacteria during winter by 28%, while ciprofloxacin resistance was seen in autumn with only 15.8%. Bacterial population led to decline due to antibiotic-induced stress. The tetracycline-resistant bacteria were completely resistant to ampicillin and 66.6% were resistant to erythromycin. In Ganga river water, 53.5% of ampicillin-resistant isolates were resistant to erythromycin and sulphadiazine 93.3% were resistant to nalidixic acid. In the Gomti River water, the most common resistance gene among tetracycline resistant isolates was tetM (83.3%), followed by ampC (83.3%) in ampicillin-resistant isolates. In the Ganga River, 66.6% of bacterial isolates were found to have ampC and ermB genes. The sul1 gene was absent in all the bacterial isolates in both water samples. CONCLUSION: These findings indicate that both rivers act as reservoirs for multidrug-resistant bacteria harbouring complex resistance gene profiles.202540928717
355530.9997Antimicrobial resistance and antimicrobial resistance genes in marine bacteria from salmon aquaculture and non-aquaculture sites. Antimicrobial resistance (AR) detected by disc diffusion and antimicrobial resistance genes detected by DNA hybridization and polymerase chain reaction with amplicon sequencing were studied in 124 marine bacterial isolates from a Chilean salmon aquaculture site and 76 from a site without aquaculture 8 km distant. Resistance to one or more antimicrobials was present in 81% of the isolates regardless of site. Resistance to tetracycline was most commonly encoded by tetA and tetG; to trimethoprim, by dfrA1, dfrA5 and dfrA12; to sulfamethizole, by sul1 and sul2; to amoxicillin, by blaTEM ; and to streptomycin, by strA-strB. Integron integrase intl1 was detected in 14 sul1-positive isolates, associated with aad9 gene cassettes in two from the aquaculture site. intl2 Integrase was only detected in three dfrA1-positive isolates from the aquaculture site and was not associated with gene cassettes in any. Of nine isolates tested for conjugation, two from the aquaculture site transferred AR determinants to Escherichia coli. High levels of AR in marine sediments from aquaculture and non-aquaculture sites suggest that dispersion of the large amounts of antimicrobials used in Chilean salmon aquaculture has created selective pressure in areas of the marine environment far removed from the initial site of use of these agents.201424612265
527140.9997Occurrence 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
292150.9997Diversity of tetracycline resistance genes in bacteria from aquaculture sources in Australia. AIMS: To determine the genetic determinants responsible for tetracycline resistance in oxytetracycline resistant bacteria from aquaculture sources in Australia. METHODS AND RESULTS: Twenty of 104 (19%) isolates tested were resistant to oxytetracycline (MIC > or = 16 microg ml(-1)). Using polymerase chain reaction (PCR) amplification, one or more tet genes were detected in 15/20 (75%) isolates tested, but none were found in 5/20 (25%). tetM (50%) was the most common determinant, followed by tetE (45%), tetA (35%) and tetD (15%). Five of 12 oxytetracycline resistant isolates studied were able to transfer their R-plasmid to Escherichia coli recipients of chicken, pig and human origin. tetA, tetD and tetM were found to be transferred while tetE was not transferred. Southern hybridization and PCR were used to confirm transfer of determinants. CONCLUSIONS: Bacterial isolates from aquaculture sources in Australia harbour a variety of tetracycline resistance genes, which can be transferred to other bacteria of different origin. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacteria from aquaculture sources in Australia contribute to the resistance gene pool reservoir. The in vitro transfer of tetracycline R-plasmid from aquatic bacteria to E. coli isolates from various sources is an indication of the potential public health risk associated with these resistance determinants.200717953612
289060.9997Genetic diversity and antimicrobial resistance of Escherichia coli from Tagus estuary (Portugal). Fecal pollution of surface waters is a current world-wide public health concern and may contribute for the dissemination of antibiotic resistance. The Tagus estuary located in the south of Portugal is one of the largest wetlands in the west coast of Europe. In this study, water samples were collected from seven stations with different anthropic pressures along the estuary and evaluated for water quality indicator bacteria. Escherichia coli isolates (n=350) were typed by REP-PCR. Representatives of each REP profile (n=220) were evaluated phenotypically for resistance to 17 antibiotics and characterized in terms of phylogenetic group. Resistant isolates were screened for the presence of antibiotic resistance genes (tet(A), tet(B), sul1, sul2, qnrA, qnrB, qnrS, aacA4-cr, bla(TEM), bla(SHV), bla(CTX-M), bla(CMY-like), bla(IMP), bla(VIM)) and integrase genes (intI1 and intI2). The highest antibiotic resistance prevalence was observed for streptomycin and tetracycline followed by β-lactams and sulphonamides. Among E. coli isolates, 65.16% were resistant to at least one of the 17 antibiotics tested and approximately 19% were multiresistant. In our E. coli population phylo-groups A and D were predominant and characterized by higher prevalence of the antibiotic resistance. intI1 and intI2 genes were found in 12% of the isolates with prevalence of class 1 integrons. A strong correlation between the prevalence of integrons and multiresistance was observed. Differences in terms of antibiotic resistance between phylogenetic groups and between sampling sites were statistically significant. The results demonstrate a high prevalence of antibiotic resistance among E. coli circulating in the Tagus estuary with emphasis on the occurrence of resistance to last-resort antibiotics and on the high incidence of multiresistance.201323714246
291970.9997Occurrence of Transferable Integrons and sul and dfr Genes Among Sulfonamide-and/or Trimethoprim-Resistant Bacteria Isolated From Chilean Salmonid Farms. Salmon farming industry in Chile currently uses a significant quantity of antimicrobials to control bacterial pathologies. The main aims of this study were to investigate the presence of transferable sulfonamide- and trimethoprim-resistance genes, sul and dfr, and their association with integrons among bacteria associated to Chilean salmon farming. For this purpose, 91 Gram-negative strains resistant to sulfisoxazole and/or trimethoprim recovered from various sources of seven Chilean salmonid farms and mainly identified as belonging to the Pseudomonas genus (81.0%) were studied. Patterns of antimicrobial resistance of strains showed a high incidence of resistance to florfenicol (98.9%), erythromycin (95.6%), furazolidone (90.1%) and amoxicillin (98.0%), whereas strains exhibited minimum inhibitory concentrations (MIC(90)) values of sulfisoxazole and trimethoprim of >4,096 and >2,048 μg mL(-1), respectively. Strains were studied for their carriage of these genes by polymerase chain reaction, using specific primers, and 28 strains (30.8%) were found to carry at least one type of sul gene, mainly associated to a class 1 integron (17 strains), and identified by 16S rRNA gene sequencing as mainly belonging to the Pseudomonas genus (21 strains). Of these, 22 strains carried the sul1 gene, 3 strains carried the sul2 gene, and 3 strains carried both the sul1 and sul2 genes. Among these, 19 strains also carried the class 1 integron-integrase gene intI1, whereas the dfrA1, dfrA12 and dfrA14 genes were detected, mostly not inserted in the class 1 integron. Otherwise, the sul3 and intI2 genes were not found. In addition, the capability to transfer by conjugation these resistance determinants was evaluated in 22 selected strains, and sul and dfr genes were successfully transferred by 10 assayed strains, mainly mediated by a 10 kb plasmid, with a frequency of transfer of 1.4 × 10(-5) to 8.4 × 10(-3) transconjugant per recipient cell, and exhibiting a co-transference of resistance to florfenicol and oxytetracycline, currently the most used in Chilean salmon industry, suggesting an antibacterial co-selection phenomenon. This is the first report of the characterization and transferability of integrons as well as sul and dfr genes among bacteria associated to Chilean salmon farms, evidencing a relevant role of this environment as a reservoir of these genes.201931031727
201680.9997Antibiotic selective pressure for the maintenance of antibiotic resistant genes in coliform bacteria isolated from the aquatic environment. Coliform bacteria isolated from the aquatic environment were investigated for antibiotic susceptibility and detailed structures of class 1 integrons. A high proportion of isolates were found to be resistant to sulfamethoxazole, aminoglycosides, and beta-lactams. The 750 (53.6%) isolates were resistant to one or more of the antibiotics tested out of 1,400 coliform bacteria. Based on the MIC of antibiotics and antibiogram, 150 isolates were selected and further studied for class 1 integrons. The intI1 gene was found in 36 (24.0%) of the 150 isolates. Twelve isolates carried the gene cassettes responsible for antibiotic resistance, while no gene cassettes were found in 24 isolates. Seven different genes, dfrA5, dfrA7, dfrA12, dfrA17, aaA2, aaA5, and aad(3'), were detected in gene cassettes. The dfrA and aad genes located on class 1 integrons were responsible for resistance to trimethoprim and aminoglycosides. The remaining 24 coliform bacteria had the incomplete or non-functional class 1 integrons. These results indicated that antibiotic selective pressures may play an important role to maintain gene cassettes of class 1 integrons and in the absence of sustained antibiotic pressures, such as the aquatic environment, coliform bacteria may carry empty or non-functional class 1 integrons.200312639037
285490.9997Occurrence of antibiotic resistance genes in culturable bacteria isolated from Turkish trout farms and their local aquatic environment. Antibiotic resistance and presence of the resistance genes were investigated in the bacteria isolated from water, sediment, and fish in trout farms. A total of 9 bacterial species, particularly Escherichia coli, were isolated from the water and sediment samples, and 12 species were isolated from fish. The antimicrobial test indicated the highest resistance against sulfamethoxazole and ampicillin in coliform bacteria, and against sulfamethoxazole, imipenem, and aztreonam in known pathogenic bacteria isolated from fish. The most effective antibiotics were rifampicin, chloramphenicol, and tetracycline. The multiple antibiotic resistance index was above the critical limit for almost all of the bacteria isolated. The most common antibiotic resistance gene was ampC, followed by tetA, sul2, blaCTX-M1, and blaTEM in the coliform bacteria. At least one resistance gene was found in 70.8% of the bacteria, and 66.6% of the bacteria had 2 or more resistance genes. Approximately 36.54% of the bacteria that contain plasmids were able to transfer them to other bacteria. The plasmid-mediated transferable resistance genes were ampC, blaCTX-M1, tetA, sul2, and blaTEM. These results indicate that the aquatic environment could play an important role in the development of antibiotic resistance and the dissemination of resistance genes among bacteria.201525993887
2930100.9997Prevalence of antibiotic resistance genes in the bacterial flora of integrated fish farming environments of Pakistan and Tanzania. The use of a wide variety of antimicrobials in human and veterinary medicine, including aquaculture, has led to the emergence of antibiotic resistant pathogens. In the present study, bacteria from water, sediments, and fish were collected from fish farms in Pakistan and Tanzania with no recorded history of antibiotic use. The isolates were screened for the presence of resistance genes against various antimicrobials used in aquaculture and animal husbandry. Resistant isolates selected by disk diffusion and genotyped by Southern hybridization were further screened by polymerase chain reaction (PCR) and amplicon sequencing. The prominent resistance genes identified encoded tetracycline [tetA(A) and tetA(G)], trimethoprim [dfrA1, dfrA5, dfrA7, dfrA12, and dfrA15], amoxicillin [bla(TEM)], streptomycin [strA-strB], chloramphenicol [cat-1], and erythromycin resistance [mefA]. The int1 gene was found in more than 30% of the bacterial isolates in association with gene cassettes. MAR indices ranged from 0.2 to 1. The bla(NDM-1) gene was not identified in ertapenem resistant isolates. It is hypothesized that integrated fish farming practices utilizing domestic farm and poultry waste along with antibiotic residues from animal husbandry may have contributed to a pool of resistance genes in the aquaculture systems studied.201222823142
5253110.9997Effects 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
2862120.9997Regulation Transcriptional of Antibiotic Resistance Genes (ARGs) in Bacteria Isolated from WWTP. The incidence of antibiotics and transcriptional regulation of ARGs in isolated bacteria from wastewater needs to be explored. By HPLC, in samples of untreated wastewater, ampicillin (49.74 ± 5.70 µg/mL), chloramphenicol (0.60 ± 0.03 µg/mL), tylosin (72.95 ± 2.03 µg/mL), and oxytetracycline (0.22 ± 0.01 µg/mL) was determined. Through metagenomic analysis identified 58 bacterial species belonging to 9 phyla and at least 14 species have shown resistance to a variety of antibiotics. Twenty-two bacterial isolates were proved to be resistant to fifteen antibiotics of new generation and used in medical research to combat infectious diseases. Fourteen strains were shown to harbor plasmids in size ranges of 2-5 Kb, 6-10 Kb and plasmids with size greater than 10 Kb. By quantitative PCR it was possible to identify genes sul, qnr, cat1, aadA1, and sat-1 gene were shown to be present in gDNA samples from treated and untreated samples of wastewater and by relative expression analysis, differential expression of cat1, ermB, act, and tetA genes was demonstrated in strains that showed identity with Escherichia coli, Bacteroides fragilis, and Salmonella thyphi, and that were stressed with different concentrations of antibiotics. The presence of ARGs in untreated water samples, as well as in bacterial isolates, was indicative that in these habitats there are microorganisms that can resist β-lactams, aminoglycosides, tetracyclines, sulfonamides, and quinolones.202337672120
2920130.9996The tetracycline resistance determinant Tet 39 and the sulphonamide resistance gene sulII are common among resistant Acinetobacter spp. isolated from integrated fish farms in Thailand. OBJECTIVES: To determine the genetic basis for tetracycline and sulphonamide resistance and the prevalence of class I and II integrons in oxytetracycline-resistant Acinetobacter spp. from integrated fish farms in Thailand. METHODS: A total of 222 isolates were screened for tetracycline resistance genes [tet(A), tet(B), tet(H), tet(M) and tet(39)] and class II integrons by PCR. One hundred and thirty-four of these isolates were also sulphonamide resistant and these isolates were screened for sulphonamide resistance genes (sulII and sulIII) as well as class I integrons. Plasmid extraction and Southern blots with sulII and tet(39) probes were performed on selected isolates. RESULTS: The recently identified tetracycline resistance gene tet(39) was demonstrated in 75% (166/222) of oxytetracycline-resistant Acinetobacter spp. from integrated fish farms in Thailand. Isolates that were also sulfamethoxazole-resistant contained sulII (96%; 129/134) and/or sulI (14%; 19/134) (as part of class I integrons). sulII and tet(39) were located on plasmids differing in size in the isolates tested. CONCLUSIONS: The study shows tet(39) and sulII to be common resistance genes among clonally distinct Acinetobacter spp. from integrated fish farms and these bacteria may constitute reservoirs of resistance genes that may increase owing to a selective pressure caused by the use of antimicrobials in the overlaying animal production.200717095527
2018140.9996Genetic diversity of three classes of integrons in antibiotic-resistant bacteria isolated from Jiulong River in southern China. We identified antibiotic-resistant bacterial isolates from the surface waters of Jiulong River basin in southern China and determined their extent of resistance, as well as the prevalence and characterization of three classes of integrons. A phylogenetic analysis of 16S ribosomal DNA (rDNA) sequences showed that 20 genera were sampled from a total of 191 strains and the most common genus was Acinetobacter. Antimicrobial susceptibility testing revealed that the 191 isolates were all multiresistant and there were high levels of resistance to 19 antimicrobials that were tested, particularly the β-lactam, sulfonamide, amphenicol, macrolide, and rifamycin classes. Moreover, class 1 integrons were ubiquitous while only five out of 191 strains harbored class 2 integrons and no class 3 integrons were detected. The variable region of the class 1 integrons contained 30 different gene cassette arrays. Nine novel arrays were found in 65 strains, and seven strains had empty integrons. Among these 30 arrays, there were 34 different gene cassettes that included 25 resistance genes, six genes with unknown functions, two mutant transposase genes, and a new gene. The unique array dfrA1-sat2-aadA1 was detected in all five isolates carrying the class 2 integron. We found that antibiotic-resistant bacterial isolates from Jiulong River were diverse and antibiotic resistance genes associated with integrons were widespread.201525869436
2931150.9996Molecular characterization of antibiotic resistance in Escherichia coli strains from a dairy cattle farm and its surroundings. BACKGROUND: This study describes the phenotypic and genotypic characteristics of 78 genetically different Escherichia coli recovered from air and exudate samples of a dairy cattle farm and its surroundings in Spain, in order to gain insight into the flow of antimicrobial resistance through the environment and food supply. RESULTS: Antimicrobial resistance was detected in 21.8% of the 78 E. coli isolates analyzed (resistance for at least one of the 14 agents tested). The highest resistance rates were recorded for ampicillin, nalidixic acid, trimethoprim/sulfamethoxazole and tetracycline. The resistance genes detected were as follows (antibiotic (number of resistant strains), gene (number of strains)): ampicillin (9), bla(TEM-1) (6); tetracycline (15), tet(A) (7), tet(B) (4), tet(A) + tet(B) (1); chloramphenicol (5), cmlA (2), floR (2); trimethoprim/sulfamethoxazole (10), sul2 (4), sul1 (3), sul3 (2), sul1 + sul2 (1); gentamicin-tobramycin (1), ant(2″) (1). About 14% of strains showed a multidrug-resistant phenotype and, of them, seven strains carried class 1 integrons containing predominantly the dfrA1-aadA1 array. One multidrug-resistant strain was found in both inside and outside air, suggesting that the airborne spread of multidrug-resistant bacteria from the animal housing facilities to the surroundings is feasible. CONCLUSIONS: This study gives a genetic background of the antimicrobial resistance problem in a dairy cattle farm and shows that air can act as a source for dissemination of antimicrobial-resistant bacteria. © 2016 Society of Chemical Industry.201726969806
2735160.9996Insight into the Antibiotic Resistance of Bacteria Isolated from Popular Aquatic Products Collected in Zhejiang, China. The present study was aimed to obtain a close insight into the distribution and diversity of antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs) among the aquatic products collected in Zhejiang, China. A total of 136 presumptive ARB picked up from six aquatic samples were classified into 22 genera and 49 species based on the 16S rDNA sequencing. Aeromonas spp., Shewanella spp., Acinetobacter spp., Myroides spp., Pseudomonas spp., and Citrobacter spp. accounted for 80% of the ARB. Among them, 109 isolates (80.15%) exhibited resistance to at least one antibiotic. Most isolates showed resistance to not only the originally selected drug but also to one to three other tested drugs. The diversity of ARB distributed in different aquatic products was significant. Furthermore, the resistance data obtained from genotypic tests were not entirely consistent with the results of the phenotypic evaluation. The genes qnrS, tetA, floR, and cmlA were frequently detected in their corresponding phenotypic resistant isolates. In contrast, the genes sul2, aac(6')-Ib, and bla (PSE) were less frequently found in the corresponding phenotypically resistant strains. The high diversity and detection rate of ARB and ARGs in aquaculture might be a significant threat to the food chains closely related to human health.202336929890
2929170.9996Occurrence of antibiotic resistance and characterization of resistance genes and integrons in Enterobacteriaceae isolated from integrated fish farms in South China. Antibiotics are still widely applied in animal husbandry to prevent diseases and used as feed additives to promote animal growth. This could result in antibiotic resistance to bacteria and antibiotic residues in animals. In this paper, Enterobacteriaceae isolated from four integrated fish farms in Zhongshan, South China were tested for antibiotic resistance, tetracycline resistance genes, sulfonamide resistance genes, and class 1 integrons. The Kirby-Bauer disk diffusion method and polymerase chain reaction (PCR) assays were carried out to test antibiotic susceptibility and resistance genes, respectively. Relatively high antibiotic resistance frequencies were found, especially for ampicillin (80%), tetracycline (52%), and trimethoprim (50%). Out of 203 Enterobacteriaceae isolates, 98.5% were resistant to one or more antibiotics tested. Multiple antibiotic resistance (MAR) was found highest in animal manures with a MAR index of 0.56. Tetracycline resistance genes (tet(A), tet(C)) and sulfonamide resistance genes (sul2) were detected in more than 50% of the isolates. The intI1 gene was found in 170 isolates (83.7%). Both classic and non-classic class 1 integrons were found. Four genes, aadA5, aadA22, dfr2, and dfrA17, were detected. To our knowledge, this is the first report for molecular characterization of antibiotic resistance genes in Enterobacteriaceae isolated from integrated fish farms in China and the first time that gene cassette array dfrA17-aadA5 has been detected in such fish farms. Results of this study indicated that fish farms may be a reservoir of highly diverse and abundant antibiotic resistant genes and gene cassettes. Integrons may play a key role in multiple antibiotic resistances posing potential health risks to the general public and aquaculture.201121975604
1363180.9996Comparison of antimicrobial resistance and molecular characterization of Escherichia coli isolates from layer breeder farms in Korea. In Korea, 4 big layer companies that possess one grandparent and 3 parent stocks are in charge of 100% of the layer chicken industry. In this study, we investigated the antimicrobial resistance of commensal 578 E. coli isolated from 20 flocks of 4-layer breeder farms (A, B, C, and D), moreover, compared the characteristics of their resistance and virulence genes. Isolates from farms B and D showed significantly higher resistance to the β-lactam antimicrobials (amoxicillin, ampicillin, and 1st-, 2nd-, and 3rd-generation cephalosporins). However, resistance to ciprofloxacin, nalidixic acid, and tetracycline was significantly higher in the isolates from farm A (P < 0.05). Interestingly, the isolates from farm C showed significantly lower resistance to most antimicrobials tested in this study. The isolates from farms B, C, and D showed the high multiple resistance to the 3 antimicrobial classes. Furthermore, the isolates from farm A showed the highest multiple resistance against the 5 classes. Among the 412 β-lactam-resistant isolates, 123 (29.9%) carried bla(TEM-1), but the distribution was significantly different among the farms from 17.5% to 51.4% (P < 0.05). Similarly, the most prevalent tetracycline resistance gene in the isolates from farms B, C, and D was tetA (50.0-77.0%); however, the isolates from farm A showed the highest prevalence in tetB (70.6%). The distribution of quinolone (qnrB, qnrD, and qnrS) and sulfonamide (su12)-resistant genes were also significantly different among the farms but that of chloramphenicol (catA1)- and aminoglycoside (aac [3]-II, and aac [6']-Ib)-resistant genes possessed no significant difference among the farms. Moreover, the isolates from farm C showed significantly higher prevalence in virulence genes (iroN, ompT, hlyF, and iss) than the other 3 farms (P < 0.05). Furthermore, the phenotypic and genotypic characteristics of E. coli isolates were significantly different among the farms, and improved management protocols are required to control of horizontal and vertical transmission of avian disease, including the dissemination of resistant bacteria in breeder flocks.202234844113
2953190.9996Diverse Mobile Genetic Elements and Conjugal Transferability of Sulfonamide Resistance Genes (sul1, sul2, and sul3) in Escherichia coli Isolates From Penaeus vannamei and Pork From Large Markets in Zhejiang, China. High prevalence rates of sulfonamide resistance genes sul1, sul2, and sul3 have been observed in Gram-negative bacteria isolated from humans, domestic animals, and aquaculture species worldwide. We investigated the distribution characteristics, location, conjugative transferability, and genetic environments of sul genes from Escherichia coli isolates collected from Penaeus vannamei and pork samples from three large markets in Zhejiang, China. The prevalence rates of sul genes in sulfonamide-resistant E. coli isolates from P. vannamei and pork samples were 90.0 and 88.6%, respectively, and the prevalence of sul1 and sul2 was significantly higher than that of sul3 (p < 0.05). Twenty-four representative sul-positive E. coli isolates were analyzed in detail. Southern blot hybridization confirmed that sul genes of E. coli isolates were located on plasmids and/or chromosomes. Transfer of resistance through conjugation was observed in all 18 E. coli isolates harboring sul genes on plasmids. Replicon typing identified seven different incompatibility groups and IncF was the dominant replicon type among sul gene-containing plasmids from both sources. PCR walking analysis indicated that 87.5% (35/40) of sul gene-related fragments carried insertion sequences (ISs) belonging to a variety of families in diverse sites, with IS26 occurring most frequently. In addition, the sul1 gene was detected mainly in fragments carrying class 1 integrons. Co-location on the same fragment with resistance genes that may contribute to the persistence and dissemination of sul1 and/or sul2 genes. The diversity of mobile genetic elements and resistance genes adjacent to sul3 was much lower than those adjacent to sul1 and sul2, especially those located in chromosomes, which reduced the transmission potential of the sul3 gene. In conclusion, combined with the results of clonal relatedness analysis by PFGE and MLST of 24 representative E. coli isolates from P. vannamei and pork samples, it showed that a small number of sul genes were vertically transmitted among E. coli from P. vannamei and that horizontal gene transfer was likely the main transmission mechanism of sul genes from both sources. Our results provide important information to better understand the risk of transmission of sul genes from seafood and meat to humans.201931428076