Isolation and Characterization of Antimicrobial-Resistant Escherichia coli from Retail Meats from Roadside Butcheries in Uganda. - Related Documents




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130301.0000Isolation and Characterization of Antimicrobial-Resistant Escherichia coli from Retail Meats from Roadside Butcheries in Uganda. Retail meats are one of the main routes for spreading antimicrobial-resistant bacteria (ARB) from livestock to humans through the food chain. In African countries, retail meats are often sold at roadside butcheries without chilling or refrigeration. Retail meats in those butcheries are suspected to be contaminated by ARB, but it was not clear. In this study, we tested for the presence of antimicrobial-resistant Escherichia coli from retail meats (n = 64) from roadside butcheries in Kampala, Uganda. The meat surfaces were swabbed and inoculated on PetriFilm SEC agar to isolate E. coli. We successfully isolated E. coli from 90.6% of these retail meat samples. We identified the phylogenetic type, antimicrobial susceptibility, and antimicrobial resistance genes prevalence between retail meat isolates (n = 89). Phylogenetic type B1 was identified from 70.8% of the retail meat isolates, suggesting that the isolates originated primarily from fecal contamination during meat processing. Tetracycline (TET)-resistant isolates with tetA and/or tetB gene(s) were the most frequently detected (28.1%), followed by ampicillin (AMP) resistance genes with bla(TEM) (15.7%,) and sulfamethoxazole-trimethoprim (SXT) resistance genes with sul2 (15.7%). No extended-spectrum beta-lactamase-producing isolates were detected. A conjugation assay showed that resistance to AMP, TET, and SXT could be simultaneously transferred to recipients. These findings suggest that antimicrobial-resistant E. coli can easily be transferred from farms to tables from retail meats obtained from roadside butcheries.202032551973
131210.9998Antimicrobial resistance profiles among Escherichia coli strains isolated from commercial and cooked foods. A total of 4330 food samples of which microbiological standard for Escherichia coli is negative in Korea were determined for the frequency of E. coli. Ninety six samples (2.2%) were positive for E. coli. Detection rate of E. coli varied significantly by food type and ranged from 0.3% to 10.9%. Seasoned raw meat (yukhoe) and cold bean-soup had the highest prevalence for E. coli (10.9%) followed by gimbap (5.2%), meat broth for cold noodle (2.9%) and sprout (2.1%). E. coli isolates (n=96) were investigated for their phenotypic and genotypic antimicrobial resistance patterns. Seventeen E. coli isolates (17.7%) were resistant to one or more antimicrobial agents tested. High rates of resistance to the following drugs were observed: tetracycline (15.6%), streptomycin (12.5%), ampicillin (10.4%), nalidixic acid (9.4%) and ticarcillin (9.4%). All ampicillin resistant isolates were screened for extended-spectrum β-lactamase (ESBL) production by the combination disk test. None of the E. coli isolates produced ESBLs. Seventeen out of 96 E. coli isolates which were resistant to at least one antibiotic were investigated by PCR for the presence of 3 classes of antimicrobial resistance genes (tetracycline, aminoglycosides and beta-lactams). The tetracycline resistance genes tetA and tetB were found in 7 and 5 isolates, respectively. The aminoglycoside resistance genes, strA/B, aphA1, aadA and aac(3)-IV were found in 9, 5, 2 and 2 isolates, respectively. The beta-lactam resistance gene, bla(TEM) was found in 7 isolates. Results of this study show that 13 E. coli isolates were multidrug resistant (to three or more antibiotics) and 12 isolates carried at least one antimicrobial resistance gene. These isolates can act as the reservoir for antimicrobial resistance genes and facilitate the dissemination of these genes to other pathogenic and commensal bacteria. Adequate intervention to reduce microbial contamination of these foods is strongly recommended.201223107506
137120.9998Presence of antimicrobial resistance in coliform bacteria from hatching broiler eggs with emphasis on ESBL/AmpC-producing bacteria. Antimicrobial resistance is recognized as one of the most important global health challenges. Broilers are an important reservoir of antimicrobial resistant bacteria in general and, more particularly, extended-spectrum β-lactamases (ESBL)/AmpC-producing Enterobacteriaceae. Since contamination of 1-day-old chicks is a potential risk factor for the introduction of antimicrobial resistant Enterobacteriaceae in the broiler production chain, the presence of antimicrobial resistant coliform bacteria in broiler hatching eggs was explored in the present study. Samples from 186 hatching eggs, collected from 11 broiler breeder farms, were inoculated on MacConkey agar with or without ceftiofur and investigated for the presence of antimicrobial resistant lactose-positive Enterobacteriaceae, particularly, ESBL/AmpC-producers. Escherichia coli and Enterobacter cloacae were obtained from the eggshells in 10 out of 11 (10/11) sampled farms. The majority of the isolates were recovered from crushed eggshells after external decontamination suggesting that these bacteria are concealed from the disinfectants in the egg shell pores. Antimicrobial resistance testing revealed that approximately 30% of the isolates showed resistance to ampicillin, tetracycline, trimethoprim and sulphonamides, while the majority of isolates were susceptible to amoxicillin-clavulanic acid, nitrofurantoin, aminoglycosides, florfenicol, neomycin and apramycin. Resistance to extended-spectrum cephalosporins was detected in eight Enterobacteriaceae isolates from five different broiler breeder farms. The ESBL phenotype was confirmed by the double disk synergy test and blaSHV-12, blaTEM-52 and blaACT-39 resistance genes were detected by PCR. This report is the first to present broiler hatching eggs as carriers and a potential source of ESBL/AmpC-producing Enterobacteriaceae for broiler chicks.201627011291
130830.9998Antimicrobial resistance genes and virulence gene encoding intimin in Escherichia coli and Enterococcus isolated from wild rabbits (Oryctolagus cuniculus) in Tunisia. The spread of antimicrobial-resistant bacteria in wildlife must be viewed as a major concern with serious implications for human and animal health. Escherichia coli and enterococcal isolates were recovered from faecal samples of 49 wild rabbits (Oryctolagus cuniculus) on specific media and were characterised using biochemical and molecular tests. For all isolates, antimicrobial susceptibility testing was performed, and resistance genes were detected by PCR. Molecular typing of isolates was carried out by pulsed-field gel-electrophoresis, and E. coli strains were also tested for the presence of intimin (eae) gene characteristic of rabbit enteropathogenic E. coli. A total of 34 E. coli and 36 enterococci [E. hirae (52.8%) and E. faecalis (47.2%)] were obtained. For E. coli, resistance to tetracycline (94%), streptomycin (62%), ciprofloxacin (47%), trimethoprim-sulphamethoxazole (35%) and chloramphenicol (6%) was observed. Resistance to third-generation cephalosporins was detected in one E. coli strain that carried the bla(CMY-2) and bla(TEM-1) genes. Class 1 integrons were detected in eight isolates. For enterococci, resistance to tetracycline (63.9%), erythromycin (30.5%), streptomycin (18.2%), and chloramphenicol (5.5%) was detected. The tet(M)+tet(L), erm(B) and ant (6)-Ia genes were identified in thirteen, seven and three resistant Enterococcus strains, respectively. Molecular typing showed a high diversity among our strains. Wild rabbits could represent a reservoir of E. coli, and enterococci carrying antimicrobial resistance genes and E. coli additionally carrying the eae gene of enteropathogenic pathotypes could both contaminate the environment. our finding seems to represent the first report of eae-positive E. coli in wild rabbits.201931842593
136940.9997Antimicrobial resistance genes in Escherichia coli isolates recovered from a commercial beef processing plantt. The goal of this study was to assess the distribution of antimicrobial resistance (AMR) genes in Escherichia coli isolates recovered from a commercial beef processing plant. A total of 123 antimicrobial-resistant E. coli isolates were used: 34 from animal hides, 10 from washed carcasses, 27 from conveyers for moving carcasses and meat, 26 from beef trimmings, and 26 from ground meat. The AMR genes for beta-lactamase (bla(CMY), bla(SHV), and bla(TEM), tetracycline (tet(A), tet(B), and tet(C)), sulfonamides (sul1, sul2, and sul3), and aminoglycoside (strA and strB) were detected by PCR assay. The distribution of tet(B), tet(C), sul1, bla(TEM), strA, and strB genes was significantly different among sample sources. E. coli isolates positive for the tet(B) gene and for both strA and strB genes together were significantly associated with hide, washed carcass, and ground meat samples, whereas sull gene was associated with washed carcass and beef trimming samples. The bla(TEM) gene was significantly associated with ground meat samples. About 50% of tetracycline-resistant E. coli isolates were positive for tet(A) (14%), tet(B) (15%), or tet(C) (21%) genes or both tet(B) and tet(C) genes together (3%). The sul2 gene or both sul1 and sul2 genes were found in 23% of sulfisoxazole-resistant E. coli isolates, whereas the sul3 gene was not found in any of the E. coli isolates tested. The majority of streptomycin-resistant E. coli isolates (76%) were positive for the strA and strB genes together. The bla(CMY), bla(TEM), and bla(SHV) genes were found in 12, 56, and 4%, respectively, of ampicillin-resistant E. coli isolates. These data suggest that E. coli isolates harboring AMR genes are widely distributed in meat processing environments and can create a pool of transferable resistance genes for pathogens. The results of this study underscore the need for effective hygienic and sanitation procedures in meat plants to reduce the risks of contamination with antimicrobial-resistant bacteria.200919517739
115650.9997Detection of qnr, aac(6')-Ib-cr and qepA genes in Escherichia coli isolated from cooked meat products in Henan, China. Antimicrobial resistance in Escherichia coli has increased in recent years in China. Antimicrobial resistant isolates and resistance genes of E. coli can be transferred to humans through the food chain and this presents a public health risk. However, few studies have investigated the prevalence of antimicrobial resistance-encoding genes in E. coli isolated from food samples in China. The aim of this study was to investigate the presence of quinolone resistance genes (QRGs) and extended-spectrum β-lactamases (ESBLs) in E. coli isolated from cooked meat products in Henan, China. A total of 75 E. coli isolates (12.1%) were detected from 620 samples. High rates of resistance to the following drugs were observed: tetracycline (56.0%), trimethoprim/sulfamethoxazole (41.3%), streptomycin (29.3%), ampicillin (26.7%) and nalidixic acid (14.7%). Of the 75 isolates, QRGs were present in 10 isolates (13.3%), with qnr and aac(6')-Ib-cr detected alone or in combination in five (6.7%) and eight isolates (10.7%). The qnr genes detected in this study included qnrS (n=3) and qnrA (n=2). The qepA gene was absent among these isolates. Three types of β-lactamase genes were identified in the five ESBL-producing E. coli isolates: blaCTX-M-1, blaCTX-M-9, and blaTEM-1. The qnrS gene was found to be co-transferred with blaCTX-M-1 and blaTEM-1 in one isolate. Our data suggest that cooked meat products may act as reservoirs for multi-resistant bacteria and facilitate the dissemination of antimicrobial resistance genes.201425036771
115760.9997Isolation and Molecular Characterization of Antimicrobial Resistant Escherichia coli from Healthy Broilers in Retail Chicken Outlets of Hotspot Cities in Southern India. E. coli is one of the first commensal bacteria to colonize the chicken gut. It may act as a source for the spread of antibiotic resistance to human via the food chain and contamination of the environment. Isolation and characterization of such E. coli from commercial broilers in retail outlets of Southern India were carried out. Eighty-three E. coli isolates (76.9%) were obtained from cloacal/meat swabs (108 samples). Phenotypically, 78.3% of isolates were ESBL producers, 69.9% were fluoroquinolone-resistant, and 6% were carbapenemase producers. Genotypically, the blaSHV, blaTEM, and blaCTX-M were present in 48.2%, 43.4%, and 10.8% of the isolates, respectively. These isolates also carried fluoroquinolone-resistant genes viz qnrB (31.3%) and qnrS (34.9%) but not carbapenemase genes. Overall, ESBL were identified in 72.3% of isolates and fluoroquinolone-resistance genes in 51.8%. Strikingly, 53% of the isolates were multidrug-resistant, with both ESBL and fluoroquinolone-resistant genes. The study revealed the presence of MDR E. coli strains in broiler meat at retail outlets indicating the potential public health risks.202540778947
115270.9997Gut Commensal Escherichia coli, a High-Risk Reservoir of Transferable Plasmid-Mediated Antimicrobial Resistance Traits. BACKGROUND: Escherichia coli (E. coli), the main human gut microorganism, is one of the evolved superbugs because of acquiring antimicrobial resistance (AMR) determinants via horizontal gene transfer (HGT). PURPOSE: This study aimed to screen isolates of gut commensal E. coli from healthy adult individuals for antimicrobial susceptibility and plasmid-mediated AMR encoding genes. METHODS: Gut commensal E. coli bacteria were isolated from fecal samples that were taken from healthy adult individuals and investigated phenotypically for their antimicrobial susceptibility against diverse classes of antimicrobials using the Kirby Bauer disc method. PCR-based molecular assays were carried out to detect diverse plasmid-carried AMR encoding genes and virulence genes of different E. coli pathotypes (eaeA, stx, ipaH, est, elt, aggR and pCVD432). The examined AMR genes were β-lactam resistance encoding genes (bla (CTX-M1), bla (TEM), bla (CMY-2)), tetracycline resistance encoding genes (tetA, tetB), sulfonamides resistance encoding genes (sul1, sulII), aminoglycoside resistance encoding genes (aac(3)-II, aac(6')-Ib-cr) and quinolones resistance encoding genes (qnrA, qnrB, qnrS). RESULTS: PCR results revealed the absence of pathotypes genes in 56 isolates that were considered gut commensal isolates. E. coli isolates showed high resistance rates against tested antimicrobial agents belonging to both β-lactams and sulfonamides (42/56, 75%) followed by quinolones (35/56, 62.5%), tetracyclines (31/56, 55.4%), while the lowest resistance rate was to aminoglycosides (24/56, 42.9%). Antimicrobial susceptibility profiles revealed that 64.3% of isolates were multidrug-resistant (MDR). High prevalence frequencies of plasmid-carried AMR genes were detected including bla (TEM) (64%) sulI (60.7%), qnrA (51.8%), aac(3)-II (37.5%), and tetA (46.4%). All isolates harbored more than one gene with the most frequent genetic profile among isolates was bla (TEM)-bla (CTX-M1-like)-qnrA-qnrB-tetA-sulI. CONCLUSION: Results are significant in the evaluation of plasmid-carried AMR genes in the human gut commensal E. coli, suggesting a potential human health risk and the necessity of strict regulation of the use of antibiotics in Egypt. Commensal E. coli bacteria may constitute a potential reservoir of AMR genes that can be transferred to other bacterial species.202235321080
101580.9997Antimicrobial-resistant and extended-spectrum β-lactamase-producing Escherichia coli in raw cow's milk. The occurrence of antimicrobial-resistant bacteria is an important public health issue. The aim of this study was the monitoring of resistant Escherichia coli in raw cow's milk with a focus on the detection of extended-spectrum β-lactamase (ESBL)-producing strains. In total, 263 samples of raw milk from 40 farms were collected and investigated in 2010 to 2013 in the Czech Republic. Detection of E. coli was performed and evaluated according to ISO 16649-2, and antibiotic resistance was screened by the disk diffusion method. The presence of E. coli was detected in 243 (92.4%) samples. In total, 270 isolates were obtained. Resistance to β-lactam (31.8%) and tetracycline (13.0%) antibiotics was detected most often and also multiresistant strains (5.5%) were observed. E. coli isolates found to be resistant to β-lactam, tetracycline, and quinolone antibiotics were assayed by PCR to detect selected genes encoding those resistance mechanisms. In isolates in which any bla genes were detected, a double-disk synergy test was performed. ESBL production was confirmed in 2 (0.7%) isolates. The genetic analysis identified the presence of the blaCTX-M gene and other resistance genes (tet(B) and qnrB). Both ESBL-positive isolates originated from the same farm and had an identical pulsed-field gel electrophoresis profile. The findings of our study indicate that milk can be a reservoir of bacteria carrying resistance genes with a potential for spreading through the food chain.201525581180
119890.9997Third-Generation Cephalosporin- and Tetracycline-Resistant Escherichia coli and Antimicrobial Resistance Genes from Metagenomes of Mink Feces and Feed. American mink (Neovison vison) is a significant source of global fur production. Except for a few studies from Denmark and Canada reporting antimicrobial resistance in bacteria isolated from clinical cases, studies from the general mink population are scarce and absent in the United States. Mink feces (n = 42) and feed (n = 8) samples obtained from a mink farm were cultured for the enumeration and detection of tetracycline-resistant (TET(r))- and third-generation cephalosporin-resistant (TGC(r))-Escherichia coli. Isolates were characterized phenotypically for their resistance to other antibiotics and genotypically for resistance genes. TET(r)E. coli were detected from 98% of feces samples (mean concentration = 6 log(10)) and from 100% of feed samples (mean concentration = 3.2 logs). Among TET(r)E. coli isolates, 44% (n = 41) of fecal- and 50% (n = 8) of feed isolates were multidrug resistant (MDR; resistance to ≥3 antimicrobial classes), and 96% (n = 49) of TET(r) isolates were positive for tet(A) and/or tet(B). TGC(r)E. coli were detected from 95% of feces and 75% of feed samples with 78% (n = 40) of fecal isolates, and all six of the feed isolates were MDR. Nearly two-thirds (65%) of the TGC(r)E. coli isolates (n = 46) were positive for bla(CMY-2); the remaining 35% were positive for bla(CTX-M,) with the bla(CTX-M-14) being the predominant (75%, n = 16) variant detected. Metagenomic DNA was extracted directly from feces and feed samples, and it was tested for 84 antimicrobial resistance genes by using quantitative polymerase chain reaction (PCR) array; selected genes were also quantified by droplet digital PCR. The genes detected from the fecal samples belonged mainly to five antimicrobial classes: macrolide-lincosamide-streptogramin B (MLS(B); 100% prevalence), TETs (88.1%), β-lactams (71.4%), aminoglycosides (66.7%), and fluoroquinolones (47.6%). β-Lactam, MLS(B), and TET resistance genes were also detected from feed samples. Our study serves as a baseline for further studies and to streamline antimicrobial use in mink production in accordance with current regulations as in food animals.202133085531
1200100.9997Virulence and Antimicrobial Resistance Patterns of Salmonella spp. Recovered From Migratory and Captive Wild Birds. BACKGROUND: Salmonella spp., especially those are resistant to extended-spectrum β-lactamase (ESBL), are considered as major concern to global health due to their emergence and dissemination. AIM: The aim of this study was to investigate the virulence and antimicrobial resistance (AMR) profile of Salmonella spp. from migratory and captive wild birds. METHOD: A total 262 faecal samples were collected, and the identification of Salmonella spp. was carried out using a standard culture and PCR as well as molecular detection of virulence and AMR genes. RESULTS: The overall prevalence of Salmonella was determined to be 30.92% (95% CI = 25.63-36.75). Migratory birds exhibited highest prevalence (38.10%), whereas wild birds in captivity showed a lower prevalence (23.40%). The agfA gene was detected at a higher rate at 24.69%. Salmonella spp. exhibited 100% resistance to tetracycline, followed by 58% ampicillin and 46% streptomycin. In addition, there was a resistance rate to ceftriaxone of 17% and to colistin sulphate of 25%. Interestingly, levofloxacin alone displayed 100% sensitivity across all isolates, while ciprofloxacin and azithromycin showed 73% and 64% sensitivity, respectively. The MAR index was 0.25 and 0.42, and 74.07% of all isolates showed multidrug resistance (MDR). It was shown that migratory and captive wild birds contained ESBL genes blaTEM (94.34% and 49.06%) and blaSHV (13.33% and 10%), respectively. Genes responsible for sulphonamide (sul1) resistance were detected in 13.33% and 79% of wild and migratory birds, respectively. CONCLUSION: Salmonella has been found in captive wild and migratory birds and could act as reservoirs for the transmission of MDR and ESBL bacteria.202439494993
1155110.9997Prevalence and antimicrobial resistance profiles of Escherichia coli isolated from free-range pigs. INTRODUCTION: Numerous studies about antimicrobial resistant Escherichia coli (E. coli) of animal origins have been conducted around the world, most of them focus on bacteria from animals raised in intensive breeding farms, but systematic studies on antimicrobial resistance in E. coli of free range animals are still lacking. METHODOLOGY: This study aimed to investigate the prevalence and antimicrobial resistance profiles of E. coli from free-range pigs in Laiwu mountainous areas, eastern China. RESULTS: Among 123 fecal samples, 123 non-duplicate E. coli were obtained with an isolation rate of 100.0％ (123/123). These E. coli showed the highest resistance rate to tetracycline (77/123, 62.6％), but all were sensitive to amoxicillin/clavulanic acid. Thirty-eight E. coli (38/123, 30.9％) showed multidrug resistance (MDR). Among 123 E. coli isolates, only 39 carried antimicrobial resistant genes detected in this study. Of these 39 isolates, blaTEM-1, blaCTX-M-14, blaCTX-M-15, qnrB, qnrD, qnrS1, floR and cfr genes were detected in 13, 9, 4, 7, 10, 7, 20, and 7 isolates, respectively. blaTEM-1 and blaCTX-M-14 genes were concomitantly detected in 6 isolates, and blaTEM, qnrB, qnrS and qnrD genes were concomitantly detected in 7 isolates. CONCLUSIONS: Free-ranging pigs may be regarded as a potential reservoir for antibiotic resistant genes.201731085827
1313120.9997Molecular epidemiology of ceftiofur-resistant Escherichia coli isolates from dairy calves. Healthy calves (n = 96, 1 to 9 weeks old) from a dairy herd in central Pennsylvania were examined each month over a five-month period for fecal shedding of ceftiofur-resistant gram-negative bacteria. Ceftiofur-resistant Escherichia coli isolates (n = 122) were characterized by antimicrobial resistance (disk diffusion and MIC), serotype, pulsed-field gel electrophoresis subtypes, beta-lactamase genes, and virulence genes. Antibiotic disk diffusion assays showed that the isolates were resistant to ampicillin (100%), ceftiofur (100%), chloramphenicol (94%), florfenicol (93%), gentamicin (89%), spectinomycin (72%), tetracycline (98%), ticarcillin (99%), and ticarcillin-clavulanic acid (99%). All isolates were multidrug resistant and displayed elevated MICs. The E. coli isolates belonged to 42 serotypes, of which O8:H25 was the predominant serotype (49.2%). Pulsed-field gel electrophoresis classified the E. coli isolates into 27 profiles. Cluster analysis showed that 77 isolates (63.1%) belonged to one unique group. The prevalence of pathogenic E. coli was low (8%). A total of 117 ceftiofur-resistant E. coli isolates (96%) possessed the bla(CMY2) gene. Based on phenotypic and genotypic characterization, the ceftiofur-resistant E. coli isolates belonged to 59 clonal types. There was no significant relationship between calf age and clonal type. The findings of this study revealed that healthy dairy calves were rapidly colonized by antibiotic-resistant strains of E. coli shortly after birth. The high prevalence of multidrug-resistant nonpathogenic E. coli in calves could be a significant source of resistance genes to other bacteria that share the same environment.200616751500
1370130.9997Risk Characterization of Antibiotic Resistance in Bacteria Isolated from Backyard, Organic, and Regular Commercial Eggs. This study was conducted to assess the risk due to antimicrobial-resistant strains of Salmonella spp., Listeria monocytogenes, and Escherichia coli isolated from the eggshell and the contents of eggs bought in markets in Valencia (Spain). Thirty-four samples from three different production styles were analyzed: standard ( n = 34), organic ( n = 16), and backyard ( n = 10) eggs. L. monocytogenes was not isolated in any style of production. Only one strain of Salmonella was isolated from standard production, which was resistant to ciprofloxacin and amoxicillin. E. coli strains were resistant in 22% of the isolates from organic production, 12.25% from standard production, and 11.23% from backyard production. In all cases, the highest resistance was observed for amoxicillin-clavulanate. None of the isolates from standard and backyard eggs were resistant to chloramphenicol, ciprofloxacin, gentamycin, and streptomycin, while only ceftriaxone was found to be effective against all E. coli isolates from organic eggs. β-Lactamase genes bla(TEM) , bla(SHV), and bla(CMY-2) and the resistance genes for tetracycline tetA, tetB, and tetC were tested. The most commonly detected antimicrobial resistance genes among the E. coli isolates were tetA (49.30%), bla(TEM) (47.89%), and tetB (36.62%). Overall, a maximum public health risk is associated with β-lactam antibiotics.201930794464
1151140.9997Genomic Analysis of Third Generation Cephalosporin Resistant Escherichia coli from Dairy Cow Manure. The production of extended-spectrum β-lactamases (ESBLs) conferring resistance to new derivatives of β-lactams is a major public health threat if present in pathogenic Gram-negative bacteria. The objective of this study was to characterize ceftiofur (TIO)- or cefotaxime (FOX)-resistant Escherichia coli isolated from dairy cow manure. Twenty-four manure samples were collected from four farms and incubated under anaerobic conditions for 20 weeks at 4 °C or at 25 °C. A total of 37 TIO- or FOX-resistant E. coli were isolated from two of the four farms to determine their susceptibility to 14 antibiotics. Among the 37 resistant E. coli, 10 different serotypes were identified, with O8:H1 being the predominant serotype (n = 17). Five isolates belonged to each of serotypes O9:NM and O153:H42, respectively. All 37 cephalosporin resistant isolates were multi-resistant with the most prevalent resistance spectrum being amoxicillin-clavulanic acid-ampicillin-cefoxitin-ceftiofur-ceftriaxone-chloramphenicol-streptomycin-sulfisoxazole-tetracycline-trimethoprim-sulfamethoxazole. The genomes of 18 selected isolates were then sequenced and compared to 14 selected human pathogenic E. coli reference genomes obtained from public repositories using different bioinformatics approaches. As expected, all 18 sequenced isolates carried at least one β-lactamase bla gene: TEM-1, TEM-81, CTX-M115, CTX-M15, OXA-1, or CMY-2. Several other antibiotic resistance genes (ARGs) and virulence determinants were detected in the sequenced isolates and all of them harbored antimicrobial resistance plasmids belonging to classic Inc groups. Our results confirm the presence of diverse ESBL producing E. coli isolates in dairy cow manure stored for a short period of time. Such manure might constitute a reservoir of resistance and virulence genes for other bacteria that share the same environment.201729149094
1377150.9997Characterization and Horizontal Transfer of Antimicrobial Resistance Genes and Integrons in Bacteria Isolated from Cooked Meat Products in China. The aim of this study was to investigate antimicrobial resistance and the presence and transferability of corresponding resistance genes and integrons in bacteria isolated from cooked meat samples in the People's Republic of China. A total of 150 isolates (22 species belonging to 15 genera) were isolated from 49 samples. Resistance of these isolates to antimicrobials was commonly observed; 42.7, 36.0, and 25.3% of the isolates were resistant to tetracycline, streptomycin, and ampicillin, respectively. Multidrug resistance was observed in 41 (27.3%) of the isolates. Sixteen resistance genes, i.e., bla(TEM-1) and bla(CTX-M-14) (β-lactams), aac(3)-IIa (gentamicin), strA and strB (streptomycin), qnrB and qnrS (fluoroquinolone), sul1, sul2, and sul3 (sulfamethoxazole), cat1 and cat2 (chloramphenicol), and tetM, tetA, tetS, and tetB (tetracycline), were found in 54 isolates. One isolate of Pseudomonas putida carried qnrB, and sequence analysis of the PCR product revealed 96% identity to qnrB2. The qnr genes were found coresiding and were cotransferred with bla genes in two isolates. Twelve isolates were positive for the class 1 integrase gene, and four isolates carried the class 2 integrase gene. However, no class 3 integrase gene was detected. One isolate of Proteus mirabilis carried dfrA32-ereA-aadA2, and this unusual array could be transferred to Escherichia coli. Nonclassic class 1 integrons lacking qacEΔ1 and sul1 genes were found in 2 of the 12 intI1-positive isolates. Our results revealed the presence of multidrug-resistant bacteria in cooked meats and the presence and transferability of resistance genes in some isolates, suggesting that cooked meat products may act as reservoirs of drug-resistant bacteria and may facilitate the spread of resistance genes.201729148877
1378160.9997Antimicrobial resistance and resistance genes in Escherichia coli strains isolated from commercial fish and seafood. The purpose of this study was to investigate the antimicrobial resistance and to characterize the implicated genes in Escherichia coli isolated from commercial fish and seafood. Fish and seafood samples (n=2663) were collected from wholesale and retail markets in Seoul, Korea between 2005 and 2008. A total of 179 E. coli isolates (6.7%) from those samples were tested for resistance to a range of antimicrobial agents. High rates of resistance to the following drugs were observed: tetracycline (30.7%), streptomycin (12.8%), cephalothin (11.7%), ampicillin (6.7%) and ticarcillin (6.1%). No resistances to amikacin, amoxicillin/clavulanic acid and cefoxitin were observed. Seventy out of 179 isolates which were resistant to one or more drugs were investigated by PCR for the presence of 3 classes of antimicrobial resistance genes (tetracycline, aminoglycosides and beta-lactams), class 1, 2 and 3 integrons. Gene cassettes of classes 1 and 2 integrons were further characterized by amplicon sequencing. The tetracycline resistance genes tetB and tetD were found in 29 (41.4%) isolates and 14 (20%) isolates, respectively. The beta-lactam resistance gene, bla(TEM) was found in 15 (21.4%) isolates. The aminoglycoside resistance gene, aadA was found in 18 (25.7%) isolates. Class 1 integron was detected in 41.4% (n=29) of the isolates, while only 2.9% (n=2) of the isolates were positive for the presence of class 2 integron. Two different gene cassettes arrangements were identified in class 1 integron-positive isolates: dfrA12-aadA2 (1.8 kb, five isolates) and aadB-aadA2 (1.6 kb, four isolates). One isolate containing class 2 integron presented the dfrA1-sat-aadA1 gene cassette array. These data suggest that commercial fish and seafood may act as the reservoir for multi-resistant bacteria and facilitate the dissemination of the resistance genes.201222071288
2962170.9997Prevalence of antimicrobial resistance in fecal Escherichia coli and Salmonella enterica in Canadian commercial meat, companion, laboratory, and shelter rabbits (Oryctolagus cuniculus) and its association with routine antimicrobial use in commercial meat rabbits. Antimicrobial resistance (AMR) in zoonotic (e.g. Salmonella spp.), pathogenic, and opportunistic (e.g. E. coli) bacteria in animals represents a potential reservoir of antimicrobial resistant bacteria and resistance genes to bacteria infecting humans and other animals. This study evaluated the prevalence of E. coli and Salmonella enterica, and the presence of associated AMR in commercial meat, companion, research, and shelter rabbits in Canada. Associations between antimicrobial usage and prevalence of AMR in bacterial isolates were also examined in commercial meat rabbits. Culture and susceptibility testing was conducted on pooled fecal samples from weanling and adult commercial meat rabbits taken during both summer and winter months (n=100, 27 farms), and from pooled laboratory (n=14, 8 laboratory facilities), companion (n=53), and shelter (n=15, 4 shelters) rabbit fecal samples. At the facility level, E. coli was identified in samples from each commercial rabbit farm, laboratory facility, and 3 of 4 shelters, and in 6 of 53 companion rabbit fecal samples. Seventy-nine of 314 (25.2%; CI: 20.7-30.2%) E. coli isolates demonstrated resistance to >1 antimicrobial agent. At least one E. coli isolate resistant to at least one antimicrobial agent was present in samples from 55.6% of commercial farms, and from 25% of each laboratory and shelter facilities, with resistance to tetracycline being most common; no resistance was identified in companion animal samples. Salmonella enterica subsp. was identified exclusively in pooled fecal samples from commercial rabbit farms; Salmonella enterica serovar London from one farm and Salmonella enterica serovar Kentucky from another. The S. Kentucky isolate was resistant to amoxicillin/clavulanic acid, ampicillin, cefoxitin, ceftiofur, ceftriaxone, streptomycin, and tetracycline, whereas the S. London isolate was pansusceptible. Routine use of antimicrobials on commercial meat rabbit farms was not significantly associated with the presence of antimicrobial resistant E. coli or S. enterica on farms; trends towards resistance were present when resistance to specific antimicrobial classes was examined. E. coli was widely prevalent in many Canadian domestic rabbit populations, while S. enterica was rare. The prevalence of AMR in isolated bacteria was variable and most common in isolates from commercial meat rabbits (96% of the AMR isolates were from commercial meat rabbit fecal samples). Our results highlight that domestic rabbits, and particularly meat rabbits, may be carriers of phenotypically antimicrobial-resistant bacteria and AMR genes, possibly contributing to transmission of these bacteria and their genes to bacteria in humans through food or direct contact, as well as to other co-housed animal species.201729254727
2032180.9997Highly variable patterns of antimicrobial resistance in commensal Escherichia coli isolates from pigs, sympatric rodents, and flies. Antimicrobial-resistant Escherichia coli strains from pigs, sympatric rodents, and flies from two large farms in the Czech Republic with different antibiotic exposure histories were characterized based on antimicrobial resistance genes, integrons, and macrorestriction DNA profiles. Isolates of E. coli were tested for susceptibility to 12 antimicrobial agents according to the standard disk diffusion method. In resistant isolates, polymerase chain reaction was used to detect antibiotic resistance genes, integrase genes, and gene cassettes. Pulsed-field gel electrophoresis (PFGE) was used for molecular subtyping of E. coli. In farm A (long-term use of amoxicillin only), 75% (n = 198), 65% (n = 49), 11% (n = 139), and 82% (n = 177) of E. coli isolates from piglets, sows, sympatric rodents, and flies, respectively, were antibiotic resistant. In farm B (various antibiotics commonly used), 53% (n = 154), 69% (n = 98), and 54% (n = 74) of E. coli isolates from piglets, sows, and sympatric rodents, respectively, were antibiotic resistant. In both farms, the highest resistance prevalence was to tetracycline, and resistance patterns of isolates were greatly variable. Isolates with the same resistance phenotype, genes, and PFGE profile were found in pigs and flies. Isolates from rodents showed unique PFGE profiles. Close contact of sympatric rodents and flies with pigs or their products was associated with colonization of rodents and flies with resistant bacteria or transfer of resistance genes found in pig intestinal flora.200919728783
1310190.9997Antimicrobial Resistance of Escherichia fergusonii Isolated from Broiler Chickens. The objective of this study was to investigate the antibiotic resistance of Escherichia fergusonii isolated from commercial broiler chicken farms. A total of 245 isolates from cloacal and cecal samples of 28- to 36-day-old chickens were collected from 32 farms. Isolates were identified using PCR, and their susceptibility to 16 antibiotics was determined by disk diffusion assay. All isolates were susceptible to meropenem, amikacin, and ciprofloxacin. The most common resistances were against ampicillin (75.1%), streptomycin (62.9%), and tetracycline (57.1%). Of the 184 ampicillin-resistant isolates, 127 were investigated using a DNA microarray carrying 75 probes for antibiotic resistance genetic determinants. Of these 127 isolates, the β-lactamase blaCMY2, blaTEM, blaACT, blaSHV, and blaCTX-M-15 genes were detected in 120 (94.5%), 31 (24.4%), 8 (6.3%), 6 (4.7%), and 4 (3.2%) isolates, respectively. Other detected genes included those conferring resistance to aminoglycosides (aadA1, strA, strB), trimethoprims (dfrV, dfrA1), tetracyclines (tetA, tetB, tetC, tetE), and sulfonamides (sul1, sul2). Class 1 integron was found in 35 (27.6%) of the ampicillin-resistant isolates. However, our data showed that the tested E. fergusonii did not carry any carbapenemase blaOXA genes. Pulsed-field gel electrophoresis revealed that the selected ampicillin-resistant E. fergusonii isolates were genetically diverse. The present study indicates that the monitoring of antimicrobial-resistant bacteria should include enteric bacteria such as E. fergusonii, which could be a reservoir of antibiotic resistance genes. The detection of isolates harboring extended-spectrum β-lactamase genes, particularly blaCTX-M-15, in this work suggests that further investigations on the occurrence of such genes in broilers are warranted.201627296596