This study was undertaken to observe the prevalence, serogroup, avian pathogenic Escherichia coli (APEC)-associated virulence gene, randomly amplified polymorphic DNA (RAPD) pattern, and antibiotic ...resistance genes of E. coli in backyard layers and their environment in India. From the 360 samples of healthy layers and their environment, 272 (75.5%) E. coli were isolated. The majority (28.67%) of them were untypeable. Among the studied virulence genes (papC, tsh, iucC, astA), 52 (14.32%) isolates were found to possess astA, including the isolates from the drinking water of the birds (4/272, 1.47%). These strains belonged to 18 different serogroups. Most of the isolates were typeable by RAPD and they produced different patterns. Phenotypic resistance of the isolates was most frequently observed to erythromycin (95.83%), chloramphenicol (87.52%), and cotrimoxazole (78.26%). None of the isolates was found to possess extended-spectrum beta-lactamases (blaTEM, blaSHV, blaCTX-M) or quinolone resistance (qnrA) genes by PCR. The present study was the first attempt in India to assess APEC distribution in backyard poultry production. Repertorio de virulencia, caracterización y análisis de los patrones de resistencia a los antibióticos de Escherichia coli aisladas de gallinas de postura de traspatio y de su medio ambiente en la India. Este estudio se realizó para determinar la prevalencia, el serogrupo, los genes asociados a virulencia de Escherichia coli patógena para las aves (APEC), los patrones de ADN polimórfico amplificado aleatoriamente (RAPD), y los genes de resistencia a los antibióticos de E. coli en gallinas de traspatio y su ambiente en la India. De las 360 muestras de gallinas sanas y de su ambiente, se aislaron 272 cepas de E. coli (75.5%). La mayoría (28.67%) de ellas no fueron tipificables. Entre los genes de virulencia estudiados (papC, tsh, iucC, astA), se encontró que 52 aislamientos (14.32%) poseían el gene astA, incluyendo los aislamientos de agua de bebida de las aves (4/272, 1.47%). Estas cepas pertenecían a 18 serogrupos diferentes. La mayoría de los aislamientos fueron tipificables mediante RAPD y produjeron diferentes patrones. La resistencia fenotípica de los aislamientos se observó con mayor frecuencia contra eritromicina (95.83%), cloranfenicol (87.52%), y contra cotrimoxazol (78.26%). Se encontró mediante PCR que ninguno de los aislamientos poseía genes de un espectro extendido de beta-lactamasas (blaTEM, blaSHV, blaCTX-M), o resistencia contra quinolonas (qnrA). El presente estudio es el primer intento en la India para evaluar la distribución de E. coli patógena para la producción de aves de traspatio.
The present study was undertaken to detect the incidence of extended-spectrum-cephalosporinase gene possessing-Escherichia coli, its co-resistance pattern against other antimicrobials, and the clonal ...relationship of the isolates in healthy kuroiler birds. A total of 80 cloacal swabs from kuroilers were collected randomly from West Bengal, India. The use of costly antimicrobials (cephalosporins) was not practiced by farmers. Escherichia coli was isolated and identified by standard biochemical tests and 16SrRNA-PCR. All the E. coll isolates, including controls, were subjected to PCR for detection of bla.sub.CTX-M, bla.sub.TEM, bla.sub.SHV, and bla.sub.CMY-2, genes. By comparing the RAPD-banding pattern, the phylogenetic relationship among the isolates was established. All the isolates were tested for phenotypical resistance against other antibiotics. In total, 60 isolates were identified as E. coli from the kuroilers studied (n = 80). Among them, 12 (20%) isolates possessed one of the studied extended-spectrum cephalosporinase genes. Among the studied genes, bla.sub.TEM, and bla.sub.SHV, were detected in 6 (10%) and 12 (20%) E. coli isolates, respectively. None of the E. coli isolates possessed bla.sub.CTX, and bla.sub.CMY-2, In phylogenetic analysis, the strains isolated from same localities with similar genetic profile were grouped into the same cluster. Resistance of extended-spectrum cephalosporinase gene possessing E. coli isolates was observed most frequently against ampicillin/cloxacillin, co-trimoxazole, amoxyclav, piperacillin, ceftriaxone, and tetracycline. Kuroiler birds with no cephalosporin usage profile may act as a reservoir of extended-spectrum cephalosporinase gene possessing E. coli. This is the first systematic study in kuroilers, to raise the awareness of consumers regarding the possibility of transmission of antimicrobial resistant E. coli from them. Key words: E. coli, extended-spectrum cephalosporinase, India, kuroiler
Istraživanje je poduzeto da se odredi incidencija bakterije E. coli koja posjeduje gene za cefalosporinaze proširenog spektra, a istodobno je rezistentna na druge antimikrobne tvari, te klonski odnos ...izolata u zdravih kokoši pasmine kuroiler. Ukupno je bilo nasumce prikupljeno 80 obrisaka kloake spomenute pasmine u Zapadnom Bengalu u Indiji. Skupe antimikrobne tvari (cefalosporini) nisu bile rabljene za liječenje. Izolati E. coli bili su identificirani standardnim biokemijskim testovima i 16SrRNA-PCR-om. Svi izolati bili su lančanom reakcijom polimerazom (PCR) pretraženi na prisutnost gena blaCTX-M, blaTEM, blaSHV i blaCMY-2. Filogenetska srodnost izolata bila je određena RAPD-om. Otpornost svih izolata bila je fenotipski pretražena i na druge antibiotike. Ukupno je izdvojeno 60 izolata E. coli iz 80 kuroiler kokoši. Od toga je u 12 (20 %) bio dokazan jedan od istraživanih gena za cefalosporinaze proširenog spektra. Među istraživanim genima, blaTEM je bio dokazan u 6 (10 %) izolata, a blaSHV u 12 (20 %) izolata E. coli. Nijedan od izolata nije posjedovao blaCTX-M i blaCMY-2. Filogenetskom analizom ustanovljeno je da su izolati iz istih lokaliteta sa sličnim genetičkim profilom bili grupirani u istu skupinu. Posjedovanje gena za cefalosporinaze proširenog spektra bilo je najčešće dokazano za antibiotike ampicilin/kloksacilin, kotrimoksazol, amoksiklav, piperacilin, ceftriakson i tetraciklin. Perad pasmine kuroiler kojoj nikad nisu davani cefalosporini može biti rezervoar E. coli koja ima gene za cefalosporinaze proširenog spektra. To je prvo sustavno istraživanje u pasmine kuroiler od značenja za svjesnost konzumenata o mogućnosti prijenosa bakterije E. coli otporne na antimikrobne tvari.
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