Izolati E. coli koji proizvode cefalosporinaze proširenog spektra izdvojeni iz kokoši pasmine kuroiler

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. coli isolates, including controls, were subjected to PCR for detection of blaCTX-M, blaTEM, blaSHV, and blaCMY-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, blaTEM and blaSHV were detected in 6 (10%) and 12 (20%) E. coli isolates, respectively. None of the E. coli isolates possessed blaCTX-M and blaCMY-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.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