Enterobacteriaceae resistant to third-generation cephalosporins and quinolones in fresh culinary herbs imported from Southeast Asia. - Related Documents




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106301.0000Enterobacteriaceae resistant to third-generation cephalosporins and quinolones in fresh culinary herbs imported from Southeast Asia. Since multidrug resistant bacteria are frequently reported from Southeast Asia, our study focused on the occurrence of ESBL-producing Enterobacteriaceae in fresh imported herbs from Thailand, Vietnam and Malaysia. Samples were collected from fresh culinary herbs imported from Southeast Asia in which ESBL-suspected isolates were obtained by selective culturing. Analysis included identification by MALDI-TOF mass spectrometry, susceptibility testing, XbaI-PFGE, microarray, PCR and sequencing of specific ESBL genes, PCR based replicon typing (PBRT) of plasmids and Southern blot hybridization. In addition, the quinolone resistance genotype was characterized by screening for plasmid mediated quinolone resistance (PMQR) genes and mutations in the quinolone resistance determining region (QRDR) of gyrA and parC. The study encompassed fifty samples of ten batches of culinary herbs (5 samples per batch) comprising nine different herb variants. The herbs originated from Thailand (Water morning glory, Acacia and Betel leaf), Vietnam (Parsley, Asian pennywort, Houttuynia leaf and Mint) and Malaysia (Holy basil and Parsley). By selective culturing 21 cefotaxime resistant Enterobacteriaceae were retrieved. Array analysis revealed 18 isolates with ESBL genes and one isolate with solely non-ESBL beta-lactamase genes. Mutations in the ampC promoter region were determined in two isolates with PCR and sequencing. The isolates were identified as Klebsiella pneumoniae (n=9), Escherichia coli (n=6), Enterobacter cloacae complex (n=5) and Enterobacter spp. (n=1). All isolates tested were multidrug resistant. Variants of CTX-M enzymes were predominantly found followed by SHV enzymes. PMQR genes (including aac(6')-1b-cr, qnrB and qnrS) were also frequently detected. In almost all cases ESBL and quinolone resistance genes were located on the same plasmid. Imported fresh culinary herbs from Southeast Asia are a potential source for contamination of food with multidrug resistant bacteria. Because these herbs are consumed without appropriate heating, transfer to human bacteria cannot be excluded.201424607424
109410.9997Detection of plasmid-mediated quinolone resistance genes in β-lactamase-producing Escherichia coli isolates from layer hens. This study was conducted to investigate the presence of plasmid-mediated quinolone resistance (PMQR) genes in β-lactamase-producing Escherichia coli isolates from layer hens and to characterize their molecular background. Among 142 E. coli isolates, 86 (60.6%) showed multidrug resistance and 15 (10.6%) were found to be β-lactamase-producing E. coli. Extended-spectrum β-lactamase (ESBL) and plasmid-mediated AmpC (pAmpC) β-lactamase genes, blaCTX-M-14 and blaCMY-2, were identified in three and six E. coli isolates, respectively. The non-ESBL or pAmpC gene, blaTEM-1, was found in eight of the isolates. Two isolates had both genes, blaCTX-M-14 and blaTEM-1. Among the 15 β-lactamase-producing E. coli, six PMQR genes, qnrS1 (n = 3) and qnrB4 (n = 3), were identified. Among the six PMQR-positive E. coli isolates, four exhibited double amino acid exchanges at both gyrA and parC with ciprofloxacin and enrofloxacin minimum inhibitory concentrations of ≥32 and ≥16 μg/mL, respectively. Additionally, five transconjugants (33.3%) showed a transferability of β-lactamase and PMQR genes. Pulsed-field gel electrophoresis (PFGE) analysis was conducted to investigate the 15 β-lactamase-producing E. coli isolates. In PFGE, E. coli included three PFGE patterns showing the same farms and in accordance with both β-lactamase and PMQR genes and the antimicrobial resistance pattern. Layer hens may act as a reservoir of antibiotic-resistant bacteria, and the PMQR gene in β-lactamase-producing E. coli isolates from layer hens has the potential to enter the food chain. Therefore, our findings suggest that comprehensive surveillance of antimicrobial use in laying operation systems is necessary.201930496543
107320.9997Occurrence of Extended Spectrum Cephalosporin-, Carbapenem- and Colistin-Resistant Gram-Negative Bacteria in Fresh Vegetables, an Increasing Human Health Concern in Algeria. The aim of this study was to screen for extended spectrum cephalosporin-, carbapenem- and colistin-resistant Gram-negative bacteria in fresh vegetables in Batna, Algeria. A total of 400 samples of fresh vegetables were collected from different retail stores. Samples were immediately subjected to selective isolation, then the representative colonies were identified using matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS). Phenotypic and genotypic analyses were carried out in terms of species identification and relative antibiotic resistance. Transferability of the carbapenemase and mcr-bearing plasmids was verified by conjugation. The clonal relationships of carbapenemase and mcr-positive Escherichia coli isolates were studied by multi-locus sequence typing (MLST). Sixty-seven isolates were characterised and were mostly isolated from green leafy vegetables, where the dominant species identified included Citrobacter freundii, Klebsiella pneumoniae, Enterobacter cloacae, Stenotrophomona maltophilia, E. coli and Citrobacter braakii. PCR and sequencing results showed that E. coli was the bacterial species presenting the highest antibiotic resistance level in parallel to bla(TEM) (n = 16) and bla(CTX-M-15) (n = 11), which were the most detected genes. Moreover, five isolates carried carbapenemase genes, including the bla(OXA-48) and/or bla(VIM-4) genes. The mcr-1 gene was detected in two E. coli isolates. MLST analysis revealed three different E. coli sequence types: ST101 (n = 1), ST216 (n = 1) and ST2298 (n = 1). Conjugation assays confirmed the transferability of the bla(OXA-48) and mcr-1 genes. In this study we report, for the first time, the detection of the bla(OXA-48) gene in E. coli and C. braakii isolates and the bla(VIM-4) gene in vegetables. To the best of our knowledge, this is the first report on the detection of mcr-1 genes from vegetables in Algeria.202235892378
110030.9997Characterization of ESBL-producing Escherichia spp. and report of an mcr-1 colistin-resistance Escherichia fergusonni strain from minced meat in Pamplona, Colombia. Foods of animal origin are increasingly considered a source of extended spectrum β-lactamase (ESBL) producing bacteria which can disseminate throughout the food chain and become a health concern for humans. This work aimed to evaluate the occurrence of ESBL-producing Escherichia coli in 100 retail minced meat samples taken in markets in Pamplona, Colombia. A total of 19 ESBL-producing isolates were obtained, 18 identified as E. coli and one as E. fergusonii. Fifteen isolates (78.9 %) carried bla(CTX-M) and bla(TEM) genes, one (5.2 %) bla(SHV) and bla(TEM) genes, one isolate (5.2 %) carried bla(CTX-M) and one (5.2 %) bla(SHV) alone. The majority of CTX-M-positive E. coli isolates carried the bla(CTX-M-15) gene (13 isolates), being the bla(CTX-M-9), bla(CTX-M-2), and bla(CTX-M-8) (one isolate each) also detected. Two SHV-positive isolates presented the bla(SHV-5) and bla(SHV-12) allele. The isolate identified as E. fergusonii was positive for bla(CTX-M-65) gene and mcr-1 gene. Sixteen isolates (84.2 %) belonged to phylogroups A and B1 and grouped together in the phylogenetic tree obtained by MLST; phylogroups E and F were also detected. Transfer of ESBL resistance was demonstrated for the E. fergusonii isolate. Whole genome sequencing of this isolate revealed the presence of plasmids carrying additional resistance genes. This investigation showed the high prevalence of ESBL-producing E. coli in retail samples of minced meat. Also, the isolation of a strain of E. fergusonii is an additional concern, as some resistance genes are located in mobile elements, which can be transmitted to other bacteria. These evidences support the increasing public health concern considering the spreading of resistance genes through the food chain.202336931145
109840.9997Extended spectrum beta-lactamase and fluoroquinolone resistance genes among Escherichia coli and Salmonella isolates from children with diarrhea, Burkina Faso. BACKGROUND: The emergence and spread of multidrug-resistant gram-negative bacteria (MDR) has become a major public health concern worldwide. This resistance is caused by enzymes-mediated genes (i.e., extended spectrum beta-lactamases) that are common in certain Enterobacterioceae species. However, the distribution of these genes is poorly documented in Burkina Faso. This study aims to determine the prevalence and distribution of the resistant genes coding for broad spectrum beta-lactamases and quinolones in rural Burkina Faso. METHODS: Multiplex PCR assays were carried out to detect ESBL-encoding genes, including bla(OXA), bla(TEM), bla(CTX-M), bla(SHV). The assays also assessed the presence of quinolone resistance gene namely qnrA, qnrB and qnrS in the quinolone-resistance DEC and Salmonella strains. RESULTS: The Extended-Spectrum Beta-Lactamases (ESBL) resistance phenotype was reported in all the E. coli isolates (5/5). Cross-resistance phenotype to quinolones (CRQ) was shown by one Salmonella strain (1/9) and three E. coli (3/5). Cross-resistance phenotypes to fluoroquinolones (CRFQ) were harboured by one Salmonella (1/9) and carbapenemase phenotypes were detected in two E. coli strains (2/5). Whilst the bla(OXA) genes were detected in 100% (5/5) of E. coli isolates and in 33.33% (3/9) Salmonella isolates. One strain of E. coli (1/5) harbored the bla(CTX-M) gene and the qnrB gene simultaneously. CONCLUSIONS: This study identified β-lactam (bla) and quinolone resistance (qnr) genes in multidrug-resistant E. coli and Salmonella spp. in rural Burkina Faso. Our finding which highlighted the enterobacteriaceae strains resistance to β-lactams and quinolones are of high interest for adequate management of antimicrobial resistant genes outbreak in Burkina Faso.202033010801
109750.9997CTX-M-producing Escherichia coli Isolated from urban pigeons (Columba livia domestica) in Brazil. INTRODUCTION: Worldwide urban pigeons (Columba livia domestica) are an important reservoir of pathogenic and multidrug-resistant bacteria (MDR). Plasmids are key genetic elements in the dissemination of antimicrobial drug resistance in bacteria, including beta-lactams and quinolones, which are the most important classes of drugs for treatment of Enterobacteriaceae infections in human and veterinary medicine. The aim of this study was to determine the presence of Escherichia coli (E. coli) harboring plasmids containing extend-spectrum (ESBL) and pAmpC beta-lactamases, also plasmid-mediated quinolone resistance (PMQR) genes in urban pigeons from São Paulo State, Brazil. METHODOLOGY: A collection of 107 isolates of E. coli from urban pigeons from four cities was screened by antimicrobial resistance phenotypic and PCR for genes encoding ESBL, pAmpC and PMQR genes. Clonality was evaluated by ERIC-PCR. RESULTS: We found three strains positive for blaCTX-M genes. In two clonally related CTX-M-8-producing strains, the gene was associated with IncI1 plasmids. An MDR strain harboring blaCTX-M-2, the plasmid could not be transferred. No strain was positive for PMQR genes. CONCLUSION: These results indicate that CTX-M-2 and CTX-M-8-producing E. coli are present in urban pigeons, which could serve as a reservoir for ESBL-producing E. coli in Brazil.201932087078
104560.9997ESBL-Producing Enterobacter cloacae Complex and Klebsiella pneumoniae Harbouring bla(CTX-M-15) and bla(CTX-M-55) Potentially Risk the Worldwide Spread of ESBL-Producing Bacteria Through Contaminated Dried Fishery Products. The transmission of life-threatening bacteria with plasmid-mediated antibiotic resistance poses a significant challenge to public health. This study aimed to determine the presence of plasmid-mediated antibiotic resistance genes in Enterobacterales isolates obtained from dried fishery products. Eighty-one dried fishery products were purchased from Vietnamese markets. Enterobacterales were isolated using a CHROMagar Escherichia coli coliform agar containing cefotaxime or meropenem. The isolated strains were assessed for their susceptibility to 14 antibiotics using a disc diffusion assay. Extended-spectrum β-lactamase (ESBL) sub-group typing was performed based on multiplex PCR of isolated ESBL-producing strains. In addition, Enterobacter cloacae AD2-1, which showed multiple drug resistance, was subject to whole-genome sequence analysis. CTX-resistant bacteria were isolated from 22% and MEM-resistant bacteria from 27% of the Vietnamese samples. CTX-resistant bacteria were isolated from 17% and MEM-resistant bacteria from 4% of Japanese samples. Bacterial identification indicated that 98 strains were isolated, of which 29 strains of E. coli, 28 of Enterobacter cloacae complex, 19 of Staphylococcus spp., and 9 of Klebsiella pneumoniae were predominant in Vietnamese samples. Japanese samples were predominantly contaminated with E. cloacae complex. Multiplex PCR and sequencing was used to determine the presence of ESBL-related genes bla(CTX-M-15) and bla(CTX-M-55) in E. cloacae and K. pneumoniae isolates. E. cloacae AD2-1 isolated from the Vietnamese dried fish was resistant to 14 antibiotics, and approximately 300 kbp of the IncHI2 plasmid harboured multiple antibiotic resistance genes and formed an antibiotic resistance gene region. This E. cloacae is considered a risk for the spread of antibiotic resistance across countries.202541171320
109370.9997The rate of frequent co-existence of plasmid-mediated quinolone resistance (PMQR) and extended-spectrum β-lactamase (ESBL) genes in Escherichia coli isolates from retail raw chicken in South Korea. Since plasmid-encoded antibiotic resistance facilitates the emergence of antibiotic-resistant bacteria, the increasing prevalence of Escherichia coli harboring plasmid-mediated quinolone resistance (PMQR) and extended-spectrum β-lactamase (ESBL) genes is a public health concern. The objective of this study is to investigate the co-existence of PMQR and ESBL genes in E. coli isolates from retail raw chicken in South Korea. Among 67 ESBL-producing E. coli isolates from 40 retail raw chicken, more than half of them carried PMQR genes, including qnrS, aac(6')-Ib-cr, and oqxAB. The qnrS was predominantly (91.4%) detected in E. coli isolates carrying both PMQR and ESBL. The aac(6')-Ib-cr was detected in seven ESBL-producing E. coli strains, and 85.7% of the aac(6')-Ib-cr-positive strains also carried qnrS. Moreover, the strains co-harboring qnrS and aac(6')-Ib-cr exhibited increased resistance to ciprofloxacin and kanamycin. These results demonstrate that PMQR genes are frequently detected in ESBL-producing E. coli isolates from retail raw chicken in South Korea.202235646407
109080.9997Distribution of extended-spectrum cephalosporin resistance determinants in Salmonella enterica and Escherichia coli isolated from broilers in southern Japan. This study was conducted to investigate the distribution and diversity of extended-spectrum cephalosporin (ESC) resistance determinants in Salmonella enterica and Escherichia coli obtained from the same cecal samples and to provide evidence of transmission of the resistance determinants among these bacteria in broiler farms in southern Japan. Salmonella enterica and E. coli were characterized by serotyping and multilocus sequence typing, respectively. An antimicrobial susceptibility test, plasmid analysis, and identification and localization of resistance genes were performed to determine the relatedness of ESC resistance determinants among the isolates. Of 48 flocks examined, 14 had S. enterica. In total, 57 S. enterica isolates were obtained, 45 of which showed ESC resistance. Extended-spectrum cephalosporin-resistant E. coli were also obtained from all of these ESC-resistant Salmonella-positive samples. β-Lactamase genes, blaTEM-52 (38 isolates), blaCTX-M-14 (1 isolate), and blaCMY-2 (6 isolates), were carried by conjugative untypable or IncP plasmids detected in the S. enterica serovars Infantis and Manhattan. The β-lactamase genes blaCTX-M-14 (3 isolates), blaCTX-M-15 (3 isolates), blaSHV-2 (1 isolate), blaSHV-12 (2 isolates), and blaCMY-2 (32 isolates) associated with IncI1-Iγ, IncFIB, IncFIC, IncK, IncB/O, and IncY plasmids were detected in E. coli co-isolates. Restriction mapping revealed similar plasmids in Salmonella Infantis and Salmonella Manhattan and in different sequence types of E. coli. Intraspecies transmission of plasmids was suggested within S. enterica and E. coli populations, whereas interspecies transmission was not observed. This study highlights the importance of plasmids as carriers of ESC resistance determinants.201323687161
110190.9997New insights into resistance to colistin and third-generation cephalosporins of Escherichia coli in poultry, Portugal: Novel bla(CTX-M-166) and bla(ESAC) genes. The increasing incidence of intestinal colonization with extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae and Gram negative organisms that has been observed in food animals such as poultry, cattle and pigs, are suggestive that animals, food and environment are potential sources of ESBL-producing bacteria. Hence, the aim of this study was to characterized commensal E. coli obtained from healthy broiler and turkey flocks at slaughter for the presence of penicillinases-, ESBL-, extended-spectrum AmpC (ESAC)-, plasmid-mediated quinolone resistance- and MCR-encoding genes. Study of clonal relatedness showed genetic diversity among CTX-M-type, SHV-12 and TEM-52 producing isolates with human isolates of the same type, was also assessed. We detected that eleven (5.4%, 11/202) and forty-five (2.2%, 45/185) E. coli isolates from broilers and turkeys, respectively, carried bla(ESBL) or bla(ESAC) genes and two isolates from turkeys carried mcr-1 gene. A new variant bla(CTX-M-166) was reported in a multidrug resistant isolate from a broiler flock. Overall, we detected a diversity of resistance mechanisms among E. coli from food-producing animals, all of them with high importance at a public health level.201729031106
1049100.9997Multiple Antibiotic-Resistant, Extended Spectrum-β-Lactamase (ESBL)-Producing Enterobacteria in Fresh Seafood. Members of the family Enterobacteriaceae include several human pathogens that can be acquired through contaminated food and water. In this study, the incidence of extended spectrum β-lactamase (ESBL)-producing enterobacteria was investigated in fresh seafood sold in retail markets. The ESBL-positive phenotype was detected in 169 (78.60%) isolates, with Escherichia coli being the predominant species (53), followed by Klebsiella oxytoca (27), and K. pneumoniae (23). More than 90% of the isolates were resistant to third generation cephalosporins, cefotaxime, ceftazidime, and cefpodoxime. Sixty-five percent of the isolates were resistant to the monobactam drug aztreonam, 40.82% to ertapenem, and 31.36% to meropenem. Resistance to at least five antibiotics was observed in 38.46% of the isolates. Polymerase Chain Reaction (PCR) analysis of ESBL-encoding genes detected bla(CTX), bla(SHV), and bla(TEM) genes in 76.92%, 63.3%, and 44.37% of the isolates, respectively. Multiple ESBL genes were detected in majority of the isolates. The recently discovered New Delhi metallo-β-lactamase gene (bla(NDM-1)) was detected in two ESBL⁺ isolates. Our study shows that secondary contamination of fresh seafood with enteric bacteria resistant to multiple antibiotics may implicate seafood as a potential carrier of antibiotic resistant bacteria and emphasizes an urgent need to prevent environmental contamination and dissemination of such bacteria.201728867789
1071110.9997Characterization of Beta-Lactamase and Fluoroquinolone Resistance Determinants in Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa Isolates from a Tertiary Hospital in Yola, Nigeria. Infections due to antimicrobial resistant gram-negative bacteria cause significant morbidity and mortality in sub-Saharan Africa. To elucidate the molecular epidemiology of antimicrobial resistance in gram-negative bacteria, we characterized beta-lactam and fluoroquinolone resistance determinants in Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa isolates collected from November 2017 to February 2018 (Period 1) and October 2021 to January 2022 (Period 2) in a tertiary medical center in north-eastern Nigeria. Whole genome sequencing (WGS) was used to identify sequence types and resistance determinants in 52 non-duplicate, phenotypically resistant isolates. Antimicrobial susceptibility was determined using broth microdilution and modified Kirby-Bauer disk diffusion methods. Twenty sequence types (STs) were identified among isolates from both periods using WGS, with increased strain diversity observed in Period 2. Common ESBL genes identified included bla(CTX-M), bla(SHV,) and bla(TEM) in both E. coli and K. pneumoniae. Notably, 50% of the E. coli in Period 2 harbored either bla(CTX-M-15) or bla(CTX-M-1 4) and phenotypically produced ESBLs. The bla(NDM-7) and bla(VIM-5) metallo-beta-lactamase genes were dominant in E. coli and P. aeruginosa in Period 1, but in Period 2, only K. pneumoniae contained bla(NDM-7), while bla(NDM-1) was predominant in P. aeruginosa. The overall rate of fluoroquinolone resistance was 77% in Period 1 but decreased to 47.8% in Period 2. Various plasmid-mediated quinolone resistance (PMQR) genes were identified in both periods, including aac(6')-Ib-cr, oqxA/oqxB, qnrA1, qnrB1, qnrB6, qnrB18, qnrVC1, as well as mutations in the chromosomal gyrA, parC and parE genes. One E. coli isolate in Period 2, which was phenotypically multidrug resistant, had ESBL bla(CTX-M-15,) the serine carbapenemase, bla(OXA-181) and mutations in the gyrA gene. The co-existence of beta-lactam and fluoroquinolone resistance markers observed in this study is consistent with widespread use of these antimicrobial agents in Nigeria. The presence of multidrug resistant isolates is concerning and highlights the importance of continued surveillance to support antimicrobial stewardship programs and curb the spread of antimicrobial resistance.202337999619
1104120.9996Predominance of Multidrug-Resistant Gram-Negative Bacteria Isolated from Supermarket Retail Seafood in Japan. Reports have documented antimicrobial usage in aquaculture, and the aquatic ecosystem can be considered a genetic storage site for antibiotic-resistant bacteria. This study assessed the prevalence of antimicrobial resistance (AMR) among Gram-negative bacteria recovered from retail seafood in Hiroshima, Japan. A total of 412 bacteria were isolated and screened for the presence of β-lactamases, acquired carbapenemases, and mobile colistin-resistance (mcr) genes. Forty-five (10.9%) isolates were dominated by Morganella (28%), Proteus (22%), Aeromonas (14%), Citrobacter (8%), and Escherichia (8%) and carried AMR genes. The identified AMR genes included those encoded in integrons (19), aac(6՛)-Ib (11), bla(TEM-1) (7), bla(CTX-M-like) (12), bla(CTX-M-65) (2), bla(SHV-12) (1), bla(SHV-27) (1), bla(OXA-10) (1), bla(OXA-2) (1), and mcr (2). The most common clinical resistances were against ampicillin, colistin, sulfamethoxazole/trimethoprim, tetracycline, and ciprofloxacin. Multidrug resistance (MDR) occurred in 27 (60%) AMR isolates, and multiple antibiotic resistance indices ranged from 0.2 to 0.8. A conjugation experiment showed that 10 of the 11 selected MDR strains harbored conjugable plasmids, although PCR-based replicon typing described seven strains as untypable. IncF replicon was identified in MDR extended-spectrum β-lactamase-producing Escherichia coli of the pathogenic B2 phylogroup. Our findings suggest that retail seafood harbors MDR bacteria of human interest that require strict resistance surveillance in the seafood production continuum.202338138079
943130.9996Occurrence, Antimicrobial Resistance Profile, and Characterization of Extended-Spectrum β-Lactamase-Producing Escherichia coli Isolates from Minced Meat at Local Markets in Thailand. Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli exhibits strong multidrug resistance (MDR) to ampicillin and third-generation cephalosporins. This study examined the occurrence, antimicrobial susceptibility, and molecular genetic features of ESBL-producing E. coli isolates from three commonly consumed minced meat varieties, namely pork, chicken, and beef. In total, 150 samples were collected from 10 local markets in Thailand. ESBL-producing E. coli was identified in 78 samples (52%), and minced chicken meat was most contaminated (79.17%). The isolates exhibited potential susceptibility to amikacin (96.16%) and carbapenems (91-95%). However, ESBL-producing E. coli displayed strong resistance to ampicillin and cefpodoxime (100%) and high MDR to 3-5 antibiotic classes (94.87%). Most presumptive ESBL producers harbored ESBL resistance genes (97.44%), most commonly bla(TEM) (78.21%). Indeed, our results demonstrated that raw minced meat has a high occurrence of ESBL-producing E. coli harboring ESBL resistance genes, highlighting the importance of implementation of sanitary handling practices to reduce microbial contamination in commercial meat as well as the need for consumer education on safe handling and cooking of meat products.202234941425
1078140.9996Prevalence of integrons, blaCTX-M and blaTEM resistance markers among ESBL-producing uropathogenic Escherichia coli isolates: first report of genomic blaCTX-M from India. Integrons have been observed to be frequently associated with uropathogenic bacteria. This study aimed at 1) determining the prevalence of class 1 integrons among ESBLl-producing uropathogenic Escherichia coli, and 2) analyzing resistance genes associated with different phylogenetic groups of the integron-positive isolates with special reference to bla(CTX-M) and bla(TEM). Twenty-three ESBL-producing E. coli were studied. Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) displayed 14 major patterns. Pulse field Gel electrophoresis-typing of 8 randomly selected integron-positive strains ruled out any correlation between genotype and antibiotype. Genomic DNA from 14 strains was PCR-positive for class 1 integrons, bla(CTX-M-15) and bla(TEM-1)-like genes. Integron-sequencing revealed "aadA5-dfrA17-dfrA7" as the most prevalent gene cassette. Our findings unveil the increasing role of the bla(CTX-M) genes in antibiotic resistance and emphasize on the significance of appropriate empirical treatment for Urinary tract infections. Moreover, this is the first study which reports bla(CTX-M) located on genomic DNA of bacteria from India.201121742580
1115150.9996Prevalence of extended spectrum beta lactamase and plasmid mediated quinolone resistant genes in strains of Klebsiella pneumonia, Morganella morganii, Leclercia adecarboxylata and Citrobacter freundii isolated from poultry in South Western Nigeria. A serious concern is arising on the coexistence of extended-spectrum beta-lactamase (ESBL) and plasmid mediated quinolone resistance (PMQR) producing bacteria in animal husbandry, which could be transferred to humans, especially in strains that may not be routinely screened for resistance. This study therefore tested the prevalence of ESBL and PMQR genes in selected bacteria isolated from poultry faeces. Faecal droppings of birds were collected from 11 farms in five states in South Western Nigeria. Bacteria were isolated from the samples on cefotaxime supplemented plates and identified with MALDI-TOF. The MIC was determined using VITEK system and resistance genes were detected with PCR. A total of 350 strains were isolated from different samples and selected strains were identified as 23 Klebsiella pneumonia, 12 Morganella morganii, seven Leclercia adecarboxylata and one Citrobacter freundii. All the species were resistant to gentamycin, trimethoprim/sulphamethaxole, tobramycin, piperacillin, cefotaxime and aztreonam (except Morganella morganii strains which were mostly susceptible to aztreonam). All the tested strains were susceptible to imipenem, meropenem and amikacin. All Leclercia adecarboxylata strains were resistant to ceftazidime, cefepime and fosfomycin while all Morganella morganii strains were resistant to fosfomycin, moxifloxacin and ciprofloxacin. All tested species were generally sensitive to ciprofloxacin except Morganella morganii strains which were resistant to ciprofloxacin. The resistance to ciprofloxacin, ceftazidime, cefepime, tigercylin, colistin and fosfomycin were 65%, 40%, 23%,, 7%, 33%, 48% respectively while the prevalence of SHV, TEM and CTX genes were 42%, 63%, 35% respectively. 9.3% of the isolates had the three ESBL genes, 2.33% had qnrA gene, 4.65% had qnr B gene while none had qnrS gene. The most prevalent PMQR gene is Oqxb (25.58%) while 6.98% had the qep gene. Klebsiella pneumoniae generally had both ESBL and PMQR genes. The high prevalence of extended spectrum beta-lactamase genes in the studied strains calls for caution in the use of beta lactam antibiotics in poultry feeds. This is the first report of the occurrence of extended spectrum beta-lactamase and plasmid mediated quinolone resistance genes in Morganella morganii and Leclercia adecarboxylata strains isolated from poultry faeces.201829942700
1011160.9996Mechanisms of cephalosporin resistance in indicator Escherichia coli isolated from food animals. Resistance to β-lactams is considered one of the major global problems and recently it became the most frequently studied topic in the area of antimicrobial resistance. The study was focused on phenotypic and genetic characterisation of commensal Escherichia coli (E. coli), including those producing cephalosporinases, isolated from gut flora of healthy slaughter animals. E. coli were cultured simultaneously on MacConkey agar (MCA) and cefotaxime supplemented MCA. The isolates were confirmed with ONPG and indol tube tests as well as PCR targeting uspA gene. Microbroth dilution method was applied for determination of Minimal Inhibitory Concentrations and interpreted according to EUCAST epidemiological cut-off values. Cephalosporin resistance phenotypes were defined by E-tests (BioMerieux) and relevant gene amplicons from selected strains were sequenced. A total of 298 E. coli isolates with cephalosporin resistance (ESC) found in 99 ones, were obtained from 318 cloacal or rectal swabs deriving from broilers, layers, turkeys, pigs and cattle. Both extended spectrum β-lactamase (ESBL) and ampC-cephalosporinase resistance phenotypes were noted in all tested animal species but cattle. At least one of the analysed genes was identified in 90 out of 99 cephalosporin-resistant isolates: bla(TEM) (n=44), bla(CMY) (n=38), bla(CTX-M) (n=33) and bla(SHV) (n=12). None of the phenotypes was identified in nine isolates. Sequencing of PCR products showed occurrence of ESBL-genes: bla(CTX-M-1/-61), bla(SHV-12), bla(TEM-1,-52/-92,-135) and ampC-gene bla(CMY-2). They were located on numerous and diverse plasmids and resistance transferability was proved by electroporation of bla(SHV-12) and bla(CTX-M-1/-61) located on X1 plasmids. Detection of cephalosporin resistant E. coli confirms the existence of resistance genes reservoir in farm animals and their possible spread (i.e. via IncX1 plasmids) to other bacteria including human and animal pathogens. The identified genetic background indicates on ecological aspects of selection and dissemination of cephalosporin resistance in E. coli isolated from food-producing animals rather than its potential role for public health threats.201626869096
1020170.9996Prevalence and characteristics of Escherichia coli strains producing extended-spectrum β -lactamases in slaughtered animals in the Czech Republic. Resistance of bacteria to antibiotics is a global medical problem requiring close cooperation between veterinary and human physicians. Raw materials and foods of animal origin may be not only a source of pathogenic bacteria causing alimentary tract infections but also a source of bacteria with a dangerous extent of resistance to antibiotics, potentially entering the human food chain. This article presents results of the first study in the Czech Republic detecting the presence of Enterobacteriaceae-producing extended-spectrum b -lactamases (ESBLs) in swabs collected in slaughterhouses from surfaces of healthy animal carcasses. In 2012, swabs taken from pig (n = 166) and cattle (n = 140) carcass surfaces were analyzed. In 17 % of 53 studied slaughterhouses, ESBL-producing Escherichia coli strains were isolated. ESBLs were found in 11 and 4 % of porcine and bovine samples, respectively. Swabs collected from pigs yielded 18 ESBL-producing E. coli strains. The bla genes were found to encode production of CTX-M-1 group enzymes in 16 strains, SHV in one case, and both CTX-M-1-like and TEM in another case. In swabs taken from cattle, five ESBL-producing E. coli strains were isolated. In three cases, the bla genes for CTX-M-1-like production were identified; in two cases, genes for both CTX-M-1-like and TEM production were found. The similarity/identity of ESBL-positive isolates was compared by pulsed-field gel electrophoresis. This is the first report and characterization of the presence and nature of ESBL-producing E. coli in swabs collected from surfaces of healthy pig and cattle carcasses in slaughterhouses in the Czech Republic.201324112579
1046180.9996Extended-spectrum β-lactamases producing multidrug resistance Escherichia coli, Salmonella and Klebsiella pneumoniae in pig population of Assam and Meghalaya, India. AIM: The present study was conducted to record the prevalence of extended spectrum β-lactamases (ESBLs) producing Escherichia coli, Salmonella spp., and Klebsiella pneumoniae from pig population of Assam and Meghalaya and to record the ability of the resistant bacteria to transfer the resistance genes horizontally. MATERIALS AND METHODS: Fecal samples (n=228), collected from pigs of Assam (n=99) and Meghalaya (n=129), were processed for isolation and identification of E. coli and Salmonella spp. All the isolates were tested for ESBLs production by double disc synergy test (DDST) followed by screening for ESBLs producing genes (bla(TEM), bla(SHV), bla(CTX-M), and bla(CMY)) by polymerase chain reaction (PCR). Possible transfer of resistance encoding genes between enteric bacterial species was carried out by in vitro and in vivo horizontal gene transfer (HGT) method. RESULTS: A total of 897 enteric bacteria (867 E. coli and 30 Salmonella) were isolated and identified. Altogether 25.41% isolates were confirmed as ESBL producers by DDST method. Majority of the isolates were E. coli followed by Salmonella. By PCR, 9.03% isolates were found positive for at least one of the target resistance genes. bla(SHV) was absent in all the isolates. bla(CMY) was the most prevalent gene. All the E. coli isolates from Assam were negative for bla(TEM). A total of 2.76% isolates were positive for bla(TEM) + bla(CMY). On the other hand, 0.67% isolates were positive for bla(CTX-M) + bla(CMY) genes. Only 0.33% isolates carried all the three genes. Altogether, 4.68% bacteria carried the resistance encoding genes in their plasmids. bla(TEM) gene could be successfully transferred from Salmonella (donor) to E. coli (recipient) by in vitro (5.5-5.7×10(-5)) and in vivo (6.5×10(-5) to 8.8×10(-4)) methods. In vivo method was more effective than in vitro in the transfer of resistance genes. CONCLUSION: The pig population of Assam and Meghalaya are carrying multidrug resistance and ESBLs producing E. coli and Salmonella. The isolates are also capable to transfer their resistance trait to other bacterial species by HGT. The present finding could be considered as a serious public health concern as similar trait can also be transmitted to the human commensal bacteria as well as pathogens.201830034183
1023190.9996Common presence of plasmid encoding bla(CTX-M-55) in extended-spectrum β-lactamase-producing Salmonella enterica and Escherichia coli isolates from the same edible river fish. The transmission of potentially life-threatening plasmid-mediated antibiotic-resistant bacteria poses a major threat to public health. This study aimed to determine the presence of commonly observed plasmids encoding plasmid-mediated antibiotic-resistance genes in Salmonella and Escherichia coli isolates from fishery products. Eighty river fishes were purchased from retail stores and supermarkets in Vietnam. Only Salmonella-positive fishes were used for antibiotic-resistant E. coli isolation. Salmonella serotyping was performed using Salmonella antisera. Isolated bacterial DNA was extracted, and antibiotic susceptibility, resistance genes, and replicon typing were determined. Our results showed that Salmonella was isolated from 12.5% (10/80) of the river fishes. Cefotaxime-resistant Salmonella was isolated from 3.8% (3/80) of the fishes and colistin-resistant Salmonella from 1.3% (1/80) . Salmonella serotyping revealed Potsdam, Schwarzengrund, Bardo/Newport, Give, Infantis, Kentucky, and Typhimurium. Multiplex polymerase chain reaction revealed the presence of extended-spectrum β-lactamase-related genes bla(CTX-M-55) and bla(CTX-M-65) and the colistin resistance gene mcr-1. To date, no study has reported an antibiotic-resistance plasmid present in multiple bacteria collected from the same food. Thus, horizontal transmission of antibiotic-resistance plasmids may occur at the food level.202337394527