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122300.9956Characterization of Escherichia coli virulence genes, pathotypes and antibiotic resistance properties in diarrheic calves in Iran. BACKGROUND: Calf diarrhea is a major economic concern in bovine industry all around the world. This study was carried out in order to investigate distribution of virulence genes, pathotypes, serogroups and antibiotic resistance properties of Escherichia coli isolated from diarrheic calves. RESULTS: Totally, 76.45% of 824 diarrheic fecal samples collected from Isfahan, Chaharmahal, Fars and Khuzestan provinces, Iran were positive for E. coli and all of them were also positive for cnf2, hlyA, cdtIII, f17c, lt, st, stx1, eae, ehly, stx2 and cnf1 virulence genes. Chaharmahal had the highest prevalence of STEC (84.61%), while Isfahan had the lowest (71.95%). E. coli serogroups had the highest frequency in 1-7 days old calves and winter season. Distribution of ETEC, EHEC, AEEC and NTEC pathotypes among E. coli isolates were 28.41%, 5.07%, 29.52% and 3.49%, respectively. Statistical analyses were significant for presence of bacteria between various seasons and ages. All isolates had the high resistance to penicillin (100%), streptomycin (98.25%) and tetracycline (98.09%) antibiotics. The most commonly detected resistance genes were aadA1, sul1, aac[3]-IV, CITM, and dfrA1. The most prevalent serogroup among STEC was O26. CONCLUSIONS: Our findings should raise awareness about antibiotic resistance in diarrheic calves in Iran. Clinicians should exercise caution when prescribing antibiotics.201425052999
123310.9954Prevalence, Antibiogram, and Resistance Profile of Extended-Spectrum β-Lactamase-Producing Escherichia coli Isolates from Pig Farms in Luzon, Philippines. This cross-sectional study was conducted to determine the prevalence, antibiogram, and resistance profile of extended-spectrum β-lactamase-producing Escherichia coli (ESBL-EC) isolates from healthy pigs and pig farms in Luzon, Philippines. A total of 162 rectal samples from healthy finisher and breeder pigs and boot swab samples from pig houses were collected from 54 randomly selected pig farms. Bacteria were isolated and screened using MacConkey agar plate supplemented with 1 mg/L cefotaxime. Identification of bacteria and antimicrobial susceptibility test were carried out through Vitek(®) 2 and combined disk test. PCR amplifications were carried out in all isolates targeting bla(CTX-M) and its five major groupings, bla(TEM), and bla(SHV). The farm prevalence of ESBL-EC was 57.41% (95% confidence interval [CI] = 43.21-70.77). A total of 48 (29.63%) ESBL-EC isolates were isolated from samples that showed 14 different phenotypic multidrug resistance patterns. The prevalence of bla(CTX-M) gene was 91.67% (95% CI = 80.02-97.68). All major bla(CTX-M-groups) except bla(CTX-M-25group) were detected. The bla(CTX-M-1) was the most prevalent bla(CTX-M) gene, 75.0% (95% CI = 60.40-86.36). The prevalence of bla(TEM) and bla(SHV) genes was 91.67% (95% CI = 80.02-97.68) and 60.42% (95% CI = 45.27-74.23), respectively. Coexistence of different bla(CTX-M), bla(TEM), and bla(SHV) genes was observed in 44 isolates with 20 different genotypic patterns. High prevalence, diverse antibiogram profile, and genotypic resistance pattern of ESBL-EC isolates from healthy pigs and pig farms were observed in this study that could result in possible transmission to farm workers, susceptible bacteria, and the environment.202031532307
129520.9954Phenotypic and genotypic characterisation of antimicrobial resistance in faecal bacteria from 30 Giant pandas. To study the prevalence of antimicrobial resistance in faecal bacteria from Giant pandas in China, 59 isolates were recovered from faecal pats of 30 Giant pandas. Antimicrobial susceptibility testing of the isolates was performed by the standardised disk diffusion method (Kirby-Bauer). Of the 59 study isolates, 32.20% were resistant to at least one antimicrobial and 16.95% showed multidrug-resistant phenotypes. Thirteen drug resistance genes [aph(3')-IIa, aac(6')-Ib, ant(3'')-Ia, aac(3)-IIa, sul1, sul2, sul3, tetA, tetC, tetM, cat1, floR and cmlA] were analysed using four primer sets by multiplex polymerase chain reaction (PCR). The detection frequency of the aph(3')-IIa gene was the highest (10.17%), followed by cmlA (8.47%). The genes aac(6')-Ib, sul2 and tetA were not detected. PCR products were confirmed by DNA sequence analysis. The results revealed that multidrug resistance was widely present in bacteria isolated from Giant pandas.200919168331
129630.9953Prevalence and antimicrobial resistance of Salmonellaisolates from goose farms in Northeast China. BACKGROUND: Salmonella is one of the most important enteric pathogenic bacteria that threatened poultry health. AIMS: This study aimed to investigate the prevalence and antimicrobial resistance of Salmonella isolates in goose farms. METHODS: A total of 244 cloacal swabs were collected from goose farms to detect Salmonella in Northeast China. Antimicrobial susceptibility, and resistance gene distribution of Salmonella isolates were investigated. RESULTS: Twenty-one Salmonella isolates were identified. Overall prevalence of Salmonella in the present study was 8.6%. Among the Salmonella isolates, the highest resistance frequencies belonged to amoxicillin (AMX) (85.7%), tetracycline (TET) and trimethoprim/sulfamethoxazole (SXT) (81%), followed by chloramphenicol (CHL) (76.2%), florfenicol (FLO) (71.4%), kanamycin (KAN) (47.6%), and gentamycin (GEN) (38.1%). Meanwhile, only 4.8% of the isolates were resistant to ciprofloxacin (CIP) and cefotaxime (CTX). None of the isolates was resistant to cefoperazone (CFP) and colistin B (CLB). Twenty isolates (95%) were simultaneously resistant to at least two antimicrobials. Ten resistance genes were detected among which the bla (TEM-1), cmlA, aac(6')-Ib-cr, sul1, sul2, sul3, and mcr-1.1 were the most prevalent, and presented in all 21 isolates followed by tetB (20/21), qnrB (19/21), and floR (15/21). CONCLUSION: Results indicated that Salmonella isolates from goose farms in Northeast China exhibited multi-drug resistance (MDR), harboring multiple antimicrobial resistance genes. Our results will be useful to design prevention and therapeutic strategies against Salmonella infection in goose farms.202033584841
134640.9953High prevalence of multidrug resistant Escherichia coli isolated from fresh vegetables sold by selected formal and informal traders in the most densely populated Province of South Africa. Contaminated fresh produce has increasingly been implicated in foodborne disease outbreaks. As microbiological safety surveillance in South Africa is limited, a total of 545 vegetable samples (spinach, tomato, lettuce, cucumber, and green beans) were purchased from retailers, street traders, trolley vendors and farmers' markets. Escherichia coli, coliforms and Enterobacteriaceae were enumerated and the prevalence of Escherichia coli, Salmonella spp. and Listeria monocytogenes determined. E. coli isolates were characterized phenotypically (antibiotic resistance) and genotypically (diarrheagenic virulence genes). Coliforms, E. coli and Enterobacteriaceae counts were mostly not significantly different between formal and informal markets, with exceptions noted on occasion. When compared to international standards, 90% to 98% tomatoes, 70% to 94% spinach, 82% cucumbers, 93% lettuce, and 80% green bean samples, had satisfactory (≤ 100 CFU/g) E. coli counts. Of the 545 vegetable samples analyzed, 14.86% (n = 81) harbored E. coli, predominantly from leafy green vegetables. Virulence genes (lt, st, bfpA, eagg, eaeA, stx1, stx2, and ipaH) were not detected in the E. coli isolates (n = 67) characterized, however 40.30% were multidrug-resistant. Resistance to aminoglycosides (neomycin, 73.13%; gentamycin, < 10%), penicillins (ampicillin, 38.81%; amoxicillin, 41.79%; augmentin, < 10%), sulfonamides (cotrimoxazole, 22.39%), tetracycline (19.4%), chloramphenicol (11.94%), cephalosporins (cefepime, 34.33%), and carbapenemases (imipenem, < 10%) were observed. This study highlights the need for continued surveillance of multidrug resistant foodborne pathogens in fresh produce retailed formally and informally for potential consumer health risks. PRACTICAL APPLICATION: The results indicate that the microbiological quality of different vegetables were similar per product type, regardless of being purchased from formal retailers or informal street traders, trolley vendors or farmers' markets. Although no pathogenic bacteria (diarrheagenic E. coli, Salmonella spp. or L. monocytogenes) were isolated, high levels of multidrug-resistance was observed in the generic E. coli isolates. These findings highlight the importance of microbiological quality surveillance of fresh produce in formal and informal markets, as these products can be a reservoir of multidrug resistant bacteria harboring antibiotic resistance and virulence genes, potentially impacting human health.202133294974
136550.9952The frequency of tetracycline resistance genes in Escherichia coli strains isolated from healthy and diarrheic pet birds. BACKGROUND: Pet birds have close contact to human and resistant bacteria can transfer from birds to intestinal flora of human. AIMS: This study was carried out to determine the tetracycline resistance genes in Escherichia coli strains associated with enteric problem in pet birds. METHODS: Totally, 295 cloacal swabs were collected from 195 healthy and 100 diarrheic pet birds in Isfahan province, Iran. The presence of E. coli was identified by conventional bacteriological, biochemical, and molecular examinations. The presence of tetracycline resistance genes (tetA, tetB, tetC, tetD, tetE, tetG, tetK, tetL, tetM, tetO, and tetS genes) were examined using three multiplex PCR. RESULTS: The results showed that 18.9% and 43% of cloacal samples of healthy and diarrheic pet birds contained E. coli, respectively. The mean percentage of E. coli isolated from cloacal samples of diarrheic birds was significantly higher than the healthy birds (46.6 vs 23.1%). In healthy birds, out of 37 E. coli isolates, 10 isolates were resistant to tetracycline, harboring tetA and tetB genes (3 tetA vs 7 tetB), but in the diarrheic birds, of 26 resistance E. coli, 11, 12, and 3 strains contained tetA (42.3%), tetB (46.15), and tetA+tetB (11.53%) genes. The percentage of tet genes were significantly higher in diarrheic birds than healthy birds (58.9 vs 24.0%). CONCLUSION: Both resistant genes of tetA and tetB were detected in E. coli isolates that are related with efflux pump activity. These genes can be transferred between Gram-negative bacteria and they have the potential ability to be transferred to the environment and human flora.202135126542
111460.9951Third-Generation Cephalosporin Resistance in Intrinsic Colistin-Resistant Enterobacterales Isolated from Retail Meat. Consumption of retail meat contaminated with antimicrobial-resistant (AMR) bacteria is a common route for transmitting clinically relevant resistant bacteria to humans. Here, we investigated the genotypic and phenotypic resistance profiles of intrinsic colistin-resistant (ICR) Enterobacterales isolated from retail meats. ICR Enterobacterales were isolated from 103 samples of chicken, 103 samples of pork, and 104 samples of beef purchased from retail shops in Japan, using colistin-containing media, and their antimicrobial susceptibility was examined. Serratia spp. (440 isolates) showed resistance to cefotaxime (19 isolates, 4.3%), tetracycline (15 isolates, 3.4%), and other antimicrobials (<1%). Hafnia spp. (136) showed resistance to cefotaxime (12 isolates, 8.6%), ceftazidime (four isolates, 2.9%), and tetracycline (two isolates, 1.4%). Proteus spp. (39) showed resistance to chloramphenicol (four isolates, 10.3%), sulfamethoxazole-trimethoprim (four isolates, 10.3%), cefotaxime (two isolates, 5.1%), kanamycin (two isolates, 5.1%), and gentamicin (one isolate, 2.6%). Cedecea spp. (22) were resistant to tetracycline (two isolates, 9.1%) whereas Morganella spp. (11) were resistant to tetracycline (four isolates, 36.4%) and chloramphenicol (one isolate, 9.2%). The resistance genes bla(fonA), bla(ACC), and bla(DHA) were detected in cefotaxime-resistant Serratia spp., Hafnia spp., and Morganella spp. isolates, respectively. This emergence of antimicrobial resistance in ICR Enterobacterales may pose a public health risk.202134943649
524570.9950Antimicrobial Resistance in U.S. Retail Ground Beef with and without Label Claims Regarding Antibiotic Use. ABSTRACT: Antibiotics used during food animal production account for approximately 77% of U.S. antimicrobial consumption by mass. Ground beef products labeled as raised without antibiotics (RWA) are perceived to harbor lower levels of antimicrobial-resistant bacteria than conventional (CONV) products with no label claims regarding antimicrobial use. Retail ground beef samples were obtained from six U.S. cities. Samples with an RWA or U.S. Department of Agriculture Organic claim (n = 299) were assigned to the RWA production system. Samples lacking these claims (n = 300) were assigned to the CONV production system. Each sample was cultured for the detection of five antimicrobial-resistant bacteria. Genomic DNA was isolated from each sample, and a quantitative PCR assay was used to determine the abundance of 10 antimicrobial resistance (AMR) genes. Prevalence of tetracycline-resistant Escherichia coli (CONV, 46.3%; RWA, 34.4%; P < 0.01) and erythromycin-resistant Enterococcus (CONV, 48.0%; RWA, 37.5%; P = 0.01) was higher in CONV ground beef. Salmonella was detected in 1.2% of samples. The AMR gene blaCTX-M (CONV, 4.1 log-normalized abundance; RWA, 3.8 log-normalized abundance; P < 0.01) was more abundant in CONV ground beef. The AMR genes mecA (CONV, 4.4 log-normalized abundance; RWA, 4.9 log-normalized abundance; P = 0.05), tet(A) (CONV, 3.9 log-normalized abundance; RWA, 4.5 log-normalized abundance; P < 0.01), tet(B) (CONV, 3.9 log-normalized abundance; RWA, 4.5 log-normalized abundance; P < 0.01), and tet(M) (CONV, 5.4 log-normalized abundance; RWA, 5.8 log-normalized abundance; P < 0.01) were more abundant in RWA ground beef. Although these results suggest that antimicrobial use during U.S. cattle production does not increase human exposure to antimicrobial-resistant bacteria via ground beef, quantitative microbiological risk assessments are required for authoritative determination of the human health impacts of the use of antimicrobial agents during beef production.202133302298
138880.9950Snapshot Study of Whole Genome Sequences of Escherichia coli from Healthy Companion Animals, Livestock, Wildlife, Humans and Food in Italy. Animals, humans and food are all interconnected sources of antimicrobial resistance (AMR), allowing extensive and rapid exchange of AMR bacteria and genes. Whole genome sequencing (WGS) was used to characterize 279 Escherichia coli isolates obtained from animals (livestock, companion animals, wildlife), food and humans in Italy. E. coli predominantly belonged to commensal phylogroups B1 (46.6%) and A (29%) using the original Clermont criteria. One hundred and thirty-six sequence types (STs) were observed, including different pandemic (ST69, ST95, ST131) and emerging (ST10, ST23, ST58, ST117, ST405, ST648) extraintestinal pathogenic Escherichia coli (ExPEC) lineages. Eight antimicrobial resistance genes (ARGs) and five chromosomal mutations conferring resistance to highest priority critically important antimicrobials (HP-CIAs) were identified (qnrS1, qnrB19, mcr-1, bla(CTX-M1,15,55), bla(CMY-2), gyrA/parC/parE, ampC and pmrB). Twenty-two class 1 integron arrangements in 34 strains were characterized and 11 ARGs were designated as intI1 related gene cassettes (aadA1, aadA2, aadA5, aad23, ant2_Ia, dfrA1, dfrA7, dfrA14, dfrA12, dfrA17, cmlA1). Notably, most intI1 positive strains belonged to rabbit (38%) and poultry (24%) sources. Three rabbit samples carried the mcr-1 colistin resistance gene in association with IS6 family insertion elements. Poultry meat harbored some of the most prominent ExPEC STs, including ST131, ST69, ST10, ST23, and ST117. Wildlife showed a high average number of virulence-associated genes (VAGs) (mean = 10), mostly associated with an ExPEC pathotype and some predominant ExPEC lineages (ST23, ST117, ST648) were identified.202033172096
115790.9949Isolation 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
1992100.9949Antimicrobial Resistance Genes, Cassettes, and Plasmids Present in Salmonella enterica Associated With United States Food Animals. The ability of antimicrobial resistance (AR) to transfer, on mobile genetic elements (MGEs) between bacteria, can cause the rapid establishment of multidrug resistance (MDR) in bacteria from animals, thus creating a foodborne risk to human health. To investigate MDR and its association with plasmids in Salmonella enterica, whole genome sequence (WGS) analysis was performed on 193 S. enterica isolated from sources associated with United States food animals between 1998 and 2011; 119 were resistant to at least one antibiotic tested. Isolates represented 86 serotypes and variants, as well as diverse phenotypic resistance profiles. A total of 923 AR genes and 212 plasmids were identified among the 193 strains. Every isolate contained at least one AR gene. At least one plasmid was detected in 157 isolates. Genes were identified for resistance to aminoglycosides (n = 472), β-lactams (n = 84), tetracyclines (n = 171), sulfonamides (n = 91), phenicols (n = 42), trimethoprim (n = 8), macrolides (n = 5), fosfomycin (n = 48), and rifampicin (n = 2). Plasmid replicon types detected in the isolates were A/C (n = 32), ColE (n = 76), F (n = 43), HI1 (n = 4), HI2 (n = 20), I1 (n = 62), N (n = 4), Q (n = 7), and X (n = 35). Phenotypic resistance correlated with the AR genes identified in 95.4% of cases. Most AR genes were located on plasmids, with many plasmids harboring multiple AR genes. Six antibiotic resistance cassette structures (ARCs) and one pseudo-cassette were identified. ARCs contained between one and five resistance genes (ARC1: sul2, strAB, tetAR; ARC2: aac3-iid; ARC3: aph, sph; ARC4: cmy-2; ARC5: floR; ARC6: tetB; pseudo-ARC: aadA, aac3-VIa, sul1). These ARCs were present in multiple isolates and on plasmids of multiple replicon types. To determine the current distribution and frequency of these ARCs, the public NCBI database was analyzed, including WGS data on isolates collected by the USDA Food Safety and Inspection Service (FSIS) from 2014 to 2018. ARC1, ARC4, and ARC5 were significantly associated with cattle isolates, while ARC6 was significantly associated with chicken isolates. This study revealed that a diverse group of plasmids, carrying AR genes, are responsible for the phenotypic resistance seen in Salmonella isolated from United States food animals. It was also determined that many plasmids carry similar ARCs.201931057528
1310110.9949Antimicrobial 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
1293120.9949Antibiotic resistance in faecal bacteria (Escherichia coli, Enterococcus spp.) in feral pigeons. AIMS: To determine the presence of antibiotic-resistant faecal Escherichia coli and Enterococcus spp. in feral pigeons (Columba livia forma domestica) in the Czech Republic. METHODS AND RESULTS: Cloacal swabs of feral pigeons collected in the city of Brno in 2006 were cultivated for antibiotic-resistant E. coli. Resistance genes, class 1 and 2 integrons, and gene cassettes were detected in resistant isolates by polymerase chain reaction (PCR). The samples were also cultivated for enterococci. Species status of enterococci isolates was determined using repetitive extragenic palindromic-PCR. Resistance genes were detected in resistant enterococci by PCR. E. coli isolates were found in 203 of 247 pigeon samples. Antibiotic resistance was recorded in three (1·5%, n(E. coli) =203) isolates. Using agar containing ciprofloxacin, 12 (5%, n(samples) =247) E. coli strains resistant to ciprofloxacin were isolated. No ESBL-producing E. coli isolates were detected. A total of 143 enterococci were isolated: Ent. faecalis (36 isolates), Ent. faecium (27), Ent. durans (19), Ent. hirae (17), Ent. mundtii (17), Ent. gallinarum (12), Ent. casseliflavus (12) and Ent. columbae (3). Resistance to one to four antibiotics was detected in 45 (31%) isolates. Resistances were determined by tetK, tetL, tetM, tetO, aac(6')aph(2''), ant(4')-Ia, aph(3')-IIIa, ermB, pbp5, vanA and vanC1 genes. CONCLUSIONS: Antibiotic-resistant E. coli and Enterococcus spp. occurred in feral pigeons in various prevalences. SIGNIFICANCE AND IMPACT OF THE STUDY: Feral pigeon should be considered a risk species for spreading in the environment antimicrobial resistant E. coli and enterococci.201020602656
1354130.9949The prevalence, antibiotic resistance and multilocus sequence typing of colistin-resistant bacteria isolated from Penaeus vannamei farms in earthen ponds and HDPE film-lined ponds in China. The aquaculture environment, especially the culture ponds and aquaculture products, is considered to be an important reservoir of colistin resistance genes. However, systematic investigations of colistin resistance in Penaeus vannamei farming in different culture modes are scarce. In this study, a total of 93 non-duplicated samples were collected from P. vannamei farms in five cities in China from 2019 to 2021. The prevalence, antibiotic resistance and multilocus sequence typing (MLST) of colistin-resistant bacteria were measured and analysed. The results showed that among the 1601 isolates in P. vannamei and its environmental samples, the pollution of colistin-resistant bacteria was serious (the overall prevalence was 37.3% and 28.8%, respectively), regardless of the earthen pond or high-density polyethylene (HDPE) film-lined pond. Among 533 isolates, the prevalence of mobile colistin resistance (mcr) genes, mcr-1, was the highest (60%, 320/533), followed by mcr-4 (1.5%, 8/533), mcr-8 (0.9%, 5/533), mcr-10 (0.6%, 3/533) and mcr-7 (0.4%, 2/533). The prevalence of mcr-1 in earthen ponds was significantly higher than that in HDPE film-lined ponds (67.5% vs. 49.1%, p < .001). The dominant strain carrying mcr-1 was Bacillus spp. (54.1%, 173/320), followed by Enterobacter spp. (8.1%, 26/320), Staphylococcus spp. (6.3%, 20/320) and Aeromonas spp. (5.3%, 17/320). The antibiotic resistance profiles of 173 Bacillus spp. varied among different sampling locations and culture types. These isolates were highly resistant to cefepime, ceftriaxone, trimethoprim-sulfamethoxazole and ceftiofur (>45%), and multidrug-resistant isolates were common (62.4%, 108/173). Sequence type (ST) 26 (37/66, 56%) was found to be the most prevalent ST in mcr-1-positive Bacillus cereus isolated from the aquaculture environment. In summary, our study pointed out that it is necessary to continuously monitor antibiotic usage and its residues regardless of the pond types, especially with regard to critical drugs such as colistin.202235841601
1304140.9949Serovar and sequence type distribution and phenotypic and genotypic antimicrobial resistance of Salmonella originating from pet animals in Chongqing, China. A total of 334 Salmonella isolates were recovered from 6,223 pet rectal samples collected at 50 pet clinics, 42 pet shops, 7 residential areas, and 4 plazas. Forty serovars were identified that included all strains except for one isolate that did not cluster via self-agglutination, with Salmonella Typhimurium monophasic variant, Salmonella Kentucky, Salmonella Enteritidis, Salmonella Pomona, and Salmonella Give being the predominant serovars. Fifty-one sequence types were identified among the isolates, and ST198, ST11, ST19, ST451, ST34, and ST155 were the most common. The top four dominant antimicrobials to which isolates were resistant were sulfisoxazole, ampicillin, doxycycline, and tetracycline, and 217 isolates exhibited multidrug resistance. The prevalence of β-lactamase genes in Salmonella isolates was 59.6%, and among these isolates, 185 harbored bla(TEM), followed by bla(CTX-M) (66) and bla(OXA) (10). Moreover, six PMQR genes, namely, including qnrA (4.8%), qnrB (4.2%), qnrD (0.9%), qnrS (18.9%), aac(6')-Ib-cr (16.5%), and oqxB (1.5%), were detected. QRDR mutations (76.6%) were very common in Salmonella isolates, with the most frequent mutation in parC (T57S) (47.3%). Furthermore, we detected six tetracycline resistance genes in 176 isolates, namely, tet(A) (39.5%), tet(B) (8.1%), tet(M) (7.7%), tet(D) (5.4%), tet(J) (3.3%), and tet(C) (1.8%), and three sulfonamide resistance genes in 303 isolates, namely, sul1 (84.4%), sul2 (31.1%), and sul3 (4.2%). Finally, we found 86 isolates simultaneously harboring four types of resistance genes that cotransferred 2-7 resistance genes to recipient bacteria. The frequent occurrence of antimicrobial resistance, particularly in dogs and cats, suggests that antibiotic misuse may be driving multidrug-resistant Salmonella among pets.IMPORTANCEPet-associated human salmonellosis has been reported for many years, and antimicrobial resistance in pet-associated Salmonella has become a serious public health problem and has attracted increasing attention. There are no reports of Salmonella from pets and their antimicrobial resistance in Chongqing, China. In this study, we investigated the prevalence, serovar diversity, sequence types, and antimicrobial resistance of Salmonella strains isolated from pet fecal samples in Chongqing. In addition, β-lactamase, QRDR, PMQR, tetracycline and sulfonamide resistance genes, and mutations in QRDRs in Salmonella isolates were examined. Our findings demonstrated the diversity of serovars and sequence types of Salmonella isolates. The isolates were widely resistant to antimicrobials, notably with a high proportion of multidrug-resistant strains, which highlights the potential direct or indirect transmission of multidrug-resistant Salmonella from pets to humans. Furthermore, resistance genes were widely prevalent in the isolates, and most of the resistance genes were spread horizontally between strains.202438757951
1091150.9949Co-harboring of cephalosporin (bla)/colistin (mcr) resistance genes among Enterobacteriaceae from flies in Thailand. The spreading of antimicrobial-resistant Enterobacteriaceae, especially those co-harboring plasmid-mediated cephalosporin (bla) and colistin (mcr) resistance genes, is becoming increasingly problematic. As a vector, flies carry antimicrobial-resistant bacteria (ARB) into human and livestock habitats. To investigate ARB in flies, we collected 235 flies from 27 sites (18 urban areas, five pig farms and four chicken farms) in Thailand during 2013-2015. Cefotaxime-resistant Enterobacteriaceae (CtxRE) and bla-positive CtxRE were isolated from 70 (29.8%) and 48 (20.4%) flies, respectively. In 93 bla-positive CtxRE isolates that included Escherichia coli, Enterobacter spp., and Klebsiella pneumoniae from 48 flies, the most frequent bla gene was TEM (n = 62), followed by CTX-M-55 (n = 31), CTX-M-14 (n = 26), CMY-2 (n = 24) and SHV (n = 10), and 58 isolates harbored multiple types of these genes. In addition, we detected the mcr-1 (n = 1) and mcr-3 (n = 19) genes in bla-positive CtxRE isolates from 16 flies. In conjugation experiments, 10 mcr-3- and bla-positive isolates exhibited co-transfer of mcr-3 and blaTEM-1 genes. These results suggest that a relatively high proportion of flies in Thailand carries cephalosporin-resistant Enterobacteriaceae harboring co-transmissible cephalosporin and colistin resistance genes.201830010911
1353160.9949Dissemination of antibiotic resistance genes, mobile genetic elements, and efflux genes in anthropogenically impacted riverine environments. Anthropogenically impacted surface waters are an important reservoir for multidrug-resistant bacteria and antibiotic-resistant genes. The present study aimed at MDR, ESBL, AmpC, efflux genes, and heavy metals resistance genes (HMRGs) in bacterial isolates from four Indian rivers belonging to different geo-climatic zones, by estimating the mode of resistance transmission exhibited by the resistant isolates. A total 71.27% isolates exhibited MDR trait, showing maximum resistance towards β-lactams (P = 66.49%; AMX = 59.04%), lincosamides (CD = 65.96%), glycopeptides (VAN = 25.19%; TEI = 56.91%), cephalosporins (CF = 53.72%; CXM = 30.32%) sulphonamide (COT = 43.62%; TRIM = 12.77%), followed by macrolide and tetracycline. The dfrA1 and dfrB genes were detected in total 37.5% isolates whereas; dfrA1 genes were detected in 33.34%. The sul1 gene was detected in 9.76% and sul2 gene was detected in 2.44% isolates. A total of 69.40% MDR integron positive isolates were detected with intI1and intI2 detected at 89.25% and 1.07%, respectively; encoding class 1 and class 2 integron-integrase. ESBL production was confirmed in 73.13% isolates that harboured the genes blaTEM (96.84%), blaSHV (27.37%), blaOXA (13.68%) and blaCTXM (18.95%) while the frequency of HMRGs; 52.24% (zntB), 33.58% (chrA), and 6.72% (cadD). Efflux activity was confirmed in 96.26% isolates that harbored the genes acrA (93.02%), tolC (88.37%), and acrB (86.04%). AmpC (plasmid-mediated) was detected in 20.9% of the riverine isolates. Detection of such hidden molecular modes of antibiotic resistance in the rivers is alarming that requires urgent and stringent measures to control the resistance threats.202133524742
1311170.9949Prevalence and Molecular Characterization of Antimicrobial Resistance in Escherichia coli Isolated from Raw Milk and Raw Milk Cheese in Egypt. The goal of this study was to examine antimicrobial resistance and characterize the implicated genes in 222 isolates of Escherichia coli from 187 samples of raw milk and the two most popular cheeses in Egypt. E. coli isolates were tested for susceptibility to 12 antimicrobials by a disk diffusion method. Among the 222 E. coli isolates, 66 (29.7%) were resistant to one or more antimicrobials, and half of these resistant isolates showed a multidrug resistance phenotype (resistance to at least three different drug classes). The resistance traits were observed to tetracycline (27.5%), ampicillin (18.9%), streptomycin (18.5%), sulfamethoxazole-trimethoprim (11.3%), cefotaxime (4.5%), kanamycin (4.1%), ceftazidime (3.6%), chloramphenicol (2.3%), nalidixic acid (1.8%), and ciprofloxacin (1.4%). No resistance to fosfomycin and imipenem was observed. Tetracycline resistance genes tetA, tetB, and tetD were detected in 53 isolates, 9 isolates, and 1 isolate, respectively, but tetC was not detected. Aminoglycoside resistance genes strA, strB, aadA, and aphA1 were detected in 41, 41, 11, and 9 isolates, respectively. Sulfonamide resistance genes sul1, sul2, and sul3 were detected in 7, 25, and 3 isolates, respectively. Of 42 ampicillin-resistant isolates, bla(TEM), bla(CTX-M), and bla(SHV) were detected in 40, 9, and 3 isolates, respectively, and 10 (23.8%) ampicillin-resistant isolates were found to produce extended-spectrum β-lactamase. Each bla gene of extended-spectrum β-lactamase-producing E. coli was further subtyped to be bla(CTX-M-15), bla(CTX-M-104), bla(TEM-1), and bla(SHV-12). The class 1 integron was also detected in 28 resistant isolates, and three different patterns were obtained by PCR-restriction fragment length polymorphism. Sequencing analysis of the variable region revealed that four isolates had dfrA12/orfF/aadA2, two had aadA22, and one had dfrA1/aadA1. These data suggest that antimicrobial-resistant E. coli are widely distributed in the milk production and processing environment in Egypt and may play a role in dissemination of antimicrobial resistance to other pathogenic and commensal bacteria.201829323530
1366180.9949Day-old chicks are a source of antimicrobial resistant bacteria for laying hen farms. Antimicrobial resistant bacteria are rarely detected in laying hens and the objective of this longitudinal study was to test day-old chick as a source. Four different commercial batches raised on the same farm were monitored from day-old chick to laying hens using Escherichia coli as a model. Ten colonies from each of the eight samplings per batch were tested for antimicrobial susceptibility using 14 antimicrobials. Overall (313 isolates), higher resistance percentages were detected for tetracycline (26.8%), followed by sulphonamides (16.3%), ampicillin (16.0%) and quinolones (10.9% and 9.3% for ciprofloxacin and nalidixic acid, respectively). Resistance percentages of bacteria from day-old chicks were higher than those of pullets and hens (p < 0.05) for tetracycline, sulphonamides, trimethoprim and chloramphenicol. Forty different phenotypic resistance profiles were detected, led by fully susceptible (182 isolates; 58.1%), and followed by single tetracycline (28 isolates; 8.9%) and ciprofloxacin/ nalidixic acid (11 isolates; 3.5%) profiles. By whole-genome sequencing, 17 genes and mutations of five chromosomal genes related to resistance were detected, the most frequent being tetA, bla(TEM-1B) and sul1. Using multilocus sequencing analysis, 58 different MLST types were detected, most of them only in a particular sample. The ST155 (27/142) was the most frequently detected, followed by ST10 (19/142) and ST48 (9/142). The fate on the farm of the detected E. coli populations in old-day chicks was not clear, but our data suggest that they did not remain in the predominant faecal population of pullets and laying hens.201930827391
1190190.9949Co-occurrence of mcr-1, mcr-3, mcr-7 and clinically relevant antimicrobial resistance genes in environmental and fecal samples. Multidrug-resistant bacteria harboring different antimicrobial resistance genes (ARGs) have been detected worldwide. The association of plasmid-mediated colistin resistance genes (mcr-like) and other ARGs in bacteria isolated from animals is a huge concern worldwide. Therefore, this study aimed to investigate the presence of mcr-like genes and clinically relevant ARGs as well as plasmids in samples from a zoo. Fecal and environmental (soil and water) samples were collected from a zoo and the DNA of cultivable aerobic bacteria was extracted. ARGs were screened by PCR and the plasmids were detected using the PCR-based replicon typing method. A total of 74 amplicons from 27 ARGs [mcr-1, mcr-3, mcr-7.1, bla(CTX-M-Gp1), bla(CTX-M-Gp2), bla(CTX-M-Gp9), bla(VEB), bla(PER), bla(CMY), tetA, tetB, tetC, aadA, aac(6')-Ib, aph(3')-Ia, ant(2'')-Ia, qnrA, qnrB, qnrS, oqxA, oqxB, sul1, sul2, sul3, cmlA, mefAE, ermB] and 21 amplicons from eight plasmid families (IncY, ColE-like, IncF(repB), IncFIA, IncFIB, IncHI1, IncFIC, IncP) were detected. These findings reinforce that the zoo acts as a reservoir of clinically relevant ARGs, including mcr-like, and call attention to the monitoring studies in the zoo. Therefore, to the best of our knowledge, this is the first report of the world of mcr-1, mcr-3 and mcr-7.1 in environmental samples from the zoo.202032382766