Extended spectrum beta-lactamase and fluoroquinolone resistance genes among Escherichia coli and Salmonella isolates from children with diarrhea, Burkina Faso. - Related Documents




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109801.0000Extended 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
109710.9999CTX-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
95420.9999Hospital effluents as a reservoir of beta-lactamase- and carbapenemase-producing enterobacteriaceae. The aim of this study was to determine the presence of beta-lactamase- (bla) producing Enterobacteriaceae in hospital effluent samples from two level II and III hospitals in Lima, Peru. The resistance profile of the isolated bacteria was identified and characterized using the MicroScan system for 18 antimicrobials, and the presence of extended spectrum beta-lactamases (ESBL) (blaCTX-M ,bla SHV bla TEM ,bla PER) and carbapenemases (bla KPC ,bla NDM ,bla VIM ,bla IMP) resistance genes was determined by conventional PCR. Thirty-two isolates were identified (20 Enterobacteriaceae and 12 gram-negative bacteria). All the isolated bacteria showed multidrug resistance. ESBL (bla TEM) and carbapenemase (blaKPC, blaIMP) genes were found in samples from the hospitals that we evaluated. The release of these microorganisms to public areas and the lack of treatment of the hospital effluents could be an important public health problem.202134468580
101930.9999First Report of OXA-48 and IMP Genes Among Extended-Spectrum Beta-Lactamase-Producing Escherichia coli Isolates from Diarrheic Calves in Tunisia. Antimicrobial resistance is one of the most serious threats to human and animal health. Evidence suggests that the overuse of antimicrobial agents in animal production has led to the emergence and dissemination of multidrug-resistant isolates. The objective of this study was to assess the rate of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in calf feces and to characterize their resistance genes for antibiotics like beta-lactams and colistin, but also to determine their virulence genes. Fecal samples were collected from 100 diarrheic calves in the region of Bizerte, Tunisia. After isolation, E. coli isolates were screened for antimicrobial resistance against 21 antibiotics by the disc diffusion method. Characterization of β-lactamase genes and determination of associated resistance genes were performed by polymerase chain reaction. Among 71 E. coli isolates, 26 (36.6%) strains were ESBL-producing. Most of these isolates were multidrug-resistant (92.3%) and the most prevalent beta-lactamase genes detected were bla(CTX-M) (n = 26), bla(SHV) (n = 11), and bla(TEM) (n = 8), whereas only 1 isolate carried the bla(CMY) gene. In addition, resistance to carbapenems was detected in two isolates; one of them harbored both bla(OXA-48) and bla(IMP) genes and the other isolate carried only the bla(IMP) gene. Several resistance genes were identified for the first time in Tunisia from cases of diarrheic calves. Furthermore, to the best of our knowledge, this is the first report of detection and identification of carbapenem resistance genes and virulence genes from calves in North Africa. A high occurrence of antimicrobial resistance of E. coli recovered from fecal samples of calves with diarrhea was observed, highlighting the need for prudent use of antimicrobial agents in veterinary medicine to decrease the incidence of multidrug-resistant bacteria for both animals and humans.202336695709
109640.9999Investigation of urban birds as source of β-lactamase-producing Gram-negative bacteria in Marseille city, France. BACKGROUND: We investigate here the presence of multidrug-resistant bacteria isolated from stool samples of yellow-legged gulls and chickens (n = 136) in urban parks and beaches of Marseille, France. Bacterial isolation was performed on selective media, including MacConkey agar with ceftriaxone and LBJMR medium. Antibiotic resistance genes, including extended-spectrum β-lactamases (ESBL) (i.e. bla(CTX-M), bla(TEM) and bla(SHV)), carbapenemases (bla(KPC), bla(VIM), bla(NDM), bla(OXA-23), bla(OXA-24), bla(OXA-48) and bla(OXA-58)) and colistin resistance genes (mcr-1 to mcr-5) were screened by real-time PCR and standard PCR and sequenced when found. RESULTS: Of the 136 stools samples collected, seven ESBL-producing Gram-negative bacteria (BGN) and 12 colistin-resistant Enterobacteriaceae were isolated. Among them, five ESBL-producing Escherichia coli and eight colistin-resistant Hafnia alvei strains were identified. Four bla(TEM-1) genes were detected in yellow-legged gulls and chickens. Three CTX-M-15 genes were detected in yellow-legged gulls and pigeons, and one CTX-M-1 in a yellow-legged gull. No mcr-1 to mcr-5 gene were detected in colistin-resistant isolates. Genotyping of E. coli strains revealed four different sequence types already described in humans and animals and one new sequence type. CONCLUSIONS: Urban birds, which are believed to have no contact with antibiotics appear as potential source of ESBL genes. Our findings highlight the important role of urban birds in the proliferation of multidrug-resistant bacteria and also the possible zoonotic transmission of such bacteria from wild birds to humans.201931672159
101850.9999Antimicrobial Resistance and Prevalence of Extended Spectrum β-Lactamase-Producing Escherichia coli from Dogs and Cats in Northeastern China from 2012 to 2021. (1) Background: there has been a growing concern about pet-spread bacterial zoonosis in recent years. This study aimed to investigate the trend in drug-resistance of canine Escherichia coli isolates in northeast China between 2012-2021 and the differences in drug-resistance of E. coli of different origins in 2021. (2) Methods: E. coli were isolated from feces or anal swab samples from dogs and cats, and their antibiotic susceptibility profiles and phylogenetic grouping were identified. PCR was applied on the extended spectrum β-lactamase (ESBL) E. coli for antibiotic resistance genes. (3) Results: five hundred and fifty-four E. coli isolates were detected in 869 samples (63.75%). The multidrug resistance (MDR) rates of E. coli in pet dogs showed a decreasing trend, but working dogs showed the opposite trend. Resistance genes bla(CTX-M) and bla(CTX-M+TEM) were dominant among the ESBL producers (n = 219). The consistency between the resistance phenotypes and genes was high except for fluoroquinolone-resistant ESBL E. coli. All ESBL E. coli-carrying bla(NDM) were isolated from working dogs, and one of the strains carried mcr-1 and bla(NDM-4). Phylogroup B2 was the dominant group in pet cats, and more than half of the isolates from companion cats were ESBL E. coli. (4) Conclusions: the measures taken to reduce resistance in China were beginning to bear fruit. Companion cats may be more susceptible to colonization by ESBL E. coli. The problem of resistant bacteria in working dogs and pet cats warrants concern.202236358160
109460.9999Detection 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
95370.9999Distribution of resistance genes encoding ESBLs in Enterobacteriaceae isolated from biological samples in health centers in Ouagadougou, Burkina Faso. OBJECTIVE: Resistance to antibiotics most especially third generation cephalosporins has assumed a worrisome dimension globally. Genes conferring these resistance which are mediated by enzymes known as extended spectrum beta-lactamases (ESBLs) are now wide spread among several Enterobacteriaceae species. However there is paucity of data regarding the distribution of these genes in Burkina Faso. Hence this prospective study aims to determine the prevalence and distribution of ESBL encoding genes in ESBL producing Enterobacteriaceae strains isolated from clinical samples of patients attending the three major hospitals in Ouagadougou Burkina Faso. RESULTS: ESBL-encoding genes were assayed in 187 ESBL producing Enterobacteriaceae strains. Among these isolates, the prevalence of ESBL-producing strains with blaTEM, blaSHV and blaCTX-M genes were 26.2% (49/187), 5.9% (11/187) and 40.1% (75/187) respectively. The association of ESBL encoding genes with health centers was statistically significant (p = 0.0209). Approximately 39.6% of E. coli harbored CTX-M and Klebsiella spp. 5.9%. This study demonstrates the dissemination of TEM, SHV and CTX-M genes in ESBL producing Enterobacteriaceae strains in Ouagadougou. Continuous spread of these bacteria poses great public health risk, thus increased surveillance and regulation of antibiotics use is imperative in Burkina Faso.201830005695
107380.9999Occurrence 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
104590.9999ESBL-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
990100.9999Resistance phenotype-genotype correlation and molecular epidemiology of Citrobacter, Enterobacter, Proteus, Providencia, Salmonella and Serratia that carry extended-spectrum β-lactamases with or without plasmid-mediated AmpC β-lactamase genes in Thailand. Extended-spectrum β-lactamases (ESBLs) and plasmid-mediated AmpC β-lactamases (pAmpCs) have been increasingly reported among less commonly encountered genera of Enterobacteriaceae. However, little is known regarding the genetic characteristics of resistance genes and epidemiology of these genera. Lack of accurate ESBL and pAmpC detection may adversely affect therapeutic outcomes. This study investigated resistance phenotype-genotype correlation and molecular epidemiology among six genera of Enterobacteriaceae (Citrobacter, Enterobacter, Proteus, Providencia, Salmonella and Serratia) that carried ESBL with or without pAmpC genes at a university hospital in Thailand. From a total of 562 isolates, 105 isolates (18.7%) had ESBL-positive phenotype whilst 140 isolates (24.9%) harboured one or more ESBL genes. CTX-M and TEM were common ESBL-related bla genes among these isolates. The sensitivity and specificity of ESBL phenotypic detection as opposed to ESBL gene detection were 70.7% and 98.6%, respectively. pAmpC genes were detected in 96 ESBL gene-carrying isolates (68.6%) and significantly caused false negative detection of ESBL. Molecular typing based on pulsed-field gel electrophoresis revealed several clones that may be endemic in this hospital. This study indicated a high prevalence of ESBLs and pAmpCs among less common members of the family Enterobacteriaceae in Thailand and these resistant bacteria need to be monitored.201120880563
1046110.9999Extended-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
1049120.9999Multiple 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
1074130.9999Extended-Spectrum β-Lactamase-Producing Klebsiella pneumoniae from Pharmaceutical Wastewaters in South-Western Nigeria. Emergence and spread of Klebsiella pneumoniae isolates producing extended-spectrum β-lactamases (ESBLs) present a major threat to public health. In this study, we characterized β-lactam-resistant K. pneumoniae isolates from six wastewater samples obtained from two pharmaceutical industries located in Lagos and Ogun States, Nigeria. Bacteria were isolated by using MacConkey agar; species identification and antibacterial susceptibility testing were performed by Vitek 2. Etest was used for ESBL phenotype confirmation. The presence of β-lactamase genes was investigated by PCR and sequencing. Bacterial strain typing was done by XbaI-macrorestriction and subsequent pulsed-field gel electrophoresis (PFGE) as well as multilocus sequence typing (MLST). Thirty-five bacterial species were isolated from the six samples; among them, we identified seven K. pneumoniae isolates with resistance to β-lactams and co-resistance to fluoroquinolones, aminoglycosides, and folate pathway inhibitors. The ESBL phenotype was confirmed in six K. pneumoniae isolates that harbored ESBL genes bla(CTX-M-15) (n = 5), bla(SHV-2) (n = 1), and bla(SHV-12) (n = 1). PFGE and MLST analysis revealed five clones belonging to four sequence types (ST11, ST15, ST37, ST101), and clone K. pneumoniae-ST101 was present in the wastewater samples from two different pharmaceutical industries. Additionally performed conjugation assays confirmed the location of β-lactamase genes on conjugative plasmids. This is the first confirmation of K. pneumoniae isolates producing CTX-M-15-ESBL from pharmaceutical wastewaters in Nigeria. The co-resistance observed might be a reflection of the different drugs produced by these industries. Continuous surveillance of the environmental reservoirs of multidrug-resistant bacteria is necessary to prevent their further spread.201728375698
1016140.9999Investigation of CTX-M Type Extended-Spectrum β-Lactamase, Carbapenem and Colistin Resistance in Enterobacterales Isolated From Dairy Cattle in Turkey. BACKGROUND: The increasing prevalence of antimicrobial resistance in animals, particularly the spread of multidrug-resistant Enterobacterales, poses a significant zoonotic and public health risk. OBJECTIVE: The aim of this study was to investigate extended-spectrum β-lactamase (ESBL), carbapenem and colistin resistance among Enterobacterales in faecal swabs of dairy cattle. METHODS: A total of 400 samples were cultured on Mac Conkey screening media for ESBL, carbapenem and colistin resistance. The grown Enterobacterales were identified by MALDI-TOF-MS, followed by ceftriaxone, cefotaxime and ceftazidime resistance and double disk synergy. ESBL resistance genes were identified by polymerase chain reaction (PCR) and Sanger sequencing. Bacteria grown on colistin screening media were investigated for colistin resistance by EUCAST microbroth dilution method. RESULTS: A total of 89 (22.25%) of the bacteria grown from 400 samples were identified as potential ESBL-producing Enterobacterales members. A number of 53 (59.5%) of them were identified as ESBL blaCTX-M as a result of PCR, and 10 of them were identified as blaCTX-M-15/28/36/66 as a result of sequencing. None of the samples cultured on carbapenem medium grew. A total of 18 samples grown in colistin medium were found to be colistin sensitive by broth microdilution. Genotypes were not included in the study. All isolated bacteria were identified as Escherichia coli. SOLUTION: In this study, blaCTX-M-15 and its derivatives, which are common in humans, were also found to be the predominant ESBL type in animals. Monitoring resistance in animals together with resistance in human infections may provide more important data on the spread of resistance.202540704983
1053150.9998Antimicrobial Resistance and Extended-Spectrum Beta-Lactamase Genes in Enterobacterales, Pseudomonas and Acinetobacter Isolates from the Uterus of Healthy Mares. Antibiotic-resistant bacteria are a growing concern for human and animal health. The objective of this study was to determine the antimicrobial resistance and extended-spectrum beta-lactamase genes in Enterobacterales, Pseudomonas spp. and Acinetobacter spp. isolates from the uterus of healthy mares. For this purpose, 21 mares were swabbed for samples, which were later seeded on blood agar and MacConkey agar. The isolates were identified using MALDI-TOF and the antimicrobial susceptibility test was performed using the Kirby-Bauer technique. To characterize the resistance genes, a polymerase chain reaction (PCR) scheme was performed. Of the isolates identified as Gram-negative, 68.8% were Enterobacterales, represented by E. coli, Enterobacter cloacae, Citrobacter spp., and Klebsiella pneumoniae; 28.1% belonged to the genus Acinetobacter spp.; and 3.1% to Pseudomonas aeruginosa. A 9.3% of the isolates were multidrug-resistant (MDR), presenting resistance to antibiotics from three different classes, while 18.8% presented resistance to two or more classes of different antibiotics. The diversity of three genes that code for ESBL (bla(TEM), bla(CTX-M) and bla(SHV)) was detected in 12.5% of the strains. The most frequent was bla(SHV), while bla(TEM) and bla(CTX-M) were present in Citrobacter spp. and Klebsiella pneumoniae. These results are an alarm call for veterinarians and their environment and suggest taking measures to prevent the spread of these microorganisms.202337764953
1020160.9998Prevalence 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
989170.9998Development of a Method for the Fast Detection of Extended-Spectrum β-Lactamase- and Plasmid-Mediated AmpC β-Lactamase-Producing Escherichia coli and Klebsiella pneumoniae from Dogs and Cats in the USA. Antibiotic resistance, such as resistance to beta-lactams and the development of resistance mechanisms, is associated with multifactorial phenomena and not only with the use of third-generation cephalosporins. Many methods have been recommended for the detection of ESBL and pAmpC β-lactamase production but they are very subjective and the appropriate facilities are not available in most laboratories, especially not in clinics. Therefore, for fast clinical antimicrobial selection, we need to rapidly detect ESBL- and pAmpC β-lactamase-producing bacteria using a simple method with samples containing large amounts of bacteria. For the detection of ESBL- and pAmpC phenotypes and genes, the disk diffusion test, DDST and multiplex PCR were conducted. Of the 109 samples, 99 (90.8%) samples were grown in MacConkey broth containing cephalothin, and 71 samples were grown on MacConkey agar containing ceftiofur. Of the 71 samples grown on MacConkey agar containing ceftiofur, 58 Escherichia coli and 19 Klebsiella pneumoniae isolates, in particular, harbored β-lactamase genes. Of the 38 samples that did not grow in MacConkey broth containing cephalothin or on MacConkey agar containing ceftiofur, 32 isolates were identified as E. coli, and 10 isolates were identified as K. pneumoniae; β-lactamase genes were not detected in these E. coli and K. pneumoniae isolates. Of the 78 ESBL- and pAmpC β-lactamase-producing E. coli and K. pneumoniae, 55 (70.5%) isolates carried one or more ESBL genes and 56 (71.8%) isolates carried one or more pAmpC β-lactamase genes. Our method is a fast, and low-cost tool for the screening of frequently encountered ESBL- and pAmpC β-lactamase-producing bacteria and it would assist in diagnosis and improve therapeutic treatment in animal hospitals.202336830436
991180.9998Characterization of extended-spectrum beta-lactamases in Enterobacteriaceae causing nosocomial infections in a Zagreb University Hospital. The bacteria producing extended-spectrum beta-lactamases (ESBLs) are increasingly reported. production of ESBLs by Gram-negative bacteria is the major mechanism of resistance to oxymino-cephalosporins and aztreonam. the aim of the present study was to characterize ESBLs produced by Enterobacteriaceae, collected during 2003-2005 in a University Hospital in Zagreb, and to determine the risk factors associated with nosocomial infections due to them. 76 isolates of Enterobacteriaceae were included in the study. Antibiotic susceptibility testing was performed by disk-diffusion and broth microdilution method according to CLSI. beta-lactamases were characterized by PCR and sequencing of bla(ESBL )genes. plasmids were extracted by alkaline lysis method and digested with EcoRI enzyme. Most of the strains displayed CAZ phenotype meaning a higher level of resistance to ceftazidime compared to cefotaxime and ceftriaxone. 50 strains produced SHV-ESBL, 28 tem and 8 CTX-M beta-lactamase. Sequencing of bla(SHV )genes from representative strains revealed SHV-5 beta-lactamase in 6 strains whereas sequencing of bla(CTX-M )genes identified CTX-M-3 beta-lactamase in 3 and CTX-M-15 in 5 strains. Strains were assigned to groups from A to f according to plasmid fingerprinting. The spread of SHV-5-producing strains throughout the hospital units could be due to selective pressure of ceftazidime which is widely prescribed in our hospital thus favoring survival of strains possessing a mutation at the Ambler position 240 responsible for ceftazidime and aztreonam resistance.200919567348
1100190.9998Characterization 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