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Original Article



Isolation and detection of antibiotics resistance genes of Escherichia coli from broiler farms in Sukabumi, Indonesia

Aprilia Hardiati, Safika Safika, I Wayan Teguh Wibawan, Agustin Indrawati, Fachriyan Hasmi Pasaribu.




Abstract

Objective: This study aimed to isolate and identify Escherichia coli from broiler samples from Sukabumi, Indonesia. Also, antibiogram studies of the isolated bacteria were carried out consid¬ering the detection of the antibiotic resistance genes.
Materials and Methods: Cloaca swabs (n = 45) were collected from broilers in Sukabumi, Indonesia. Isolation and identification of E. coli were carried out according to standard bacterio¬logical techniques and biochemical tests, followed by confirmation of the polymerase chain reac¬tion targeting the uspA gene. Antibiotic sensitivity test, using several antibiotics [tetracycline (TE), oxytetracycline (OT), ampicillin (AMP), gentamicin (CN), nalidixic acid (NA), ciprofloxacin (CIP), enrofloxacin (ENR), chloramphenicol, and erythromycin] was carried out following the Kirby– Bauer disk diffusion method. Detection of antibiotic resistance coding genes was carried out by PCR using specific oligonucleotide primers. Statistical analysis was carried out with one-way anal¬ysis of variance.
Results: The results showed that 55.6% (25/45) of the samples were associated with the pres¬ence of E. coli. Antibiotic sensitivity test showed that the E. coli isolates were resistant to TE (88%; 22/25), OT (88%; 22/25), AMP (100%; 25/25), CN (64%; 16/25), NA (100%; 22/25), CIP (88%; 22/25), ENR (72%; 18/25), chloramphenicol (0%; 0/25), and erythromycin (92%; 23/25). On the other hand, the antibiotic resistance coding genes were tetA (86.4%; 19/22), blaTEM (100%; 25/25), aac(3)-IV (0%; 0/16), gyrA (100%; 25/25), and ermB (13%; 3/23). It was found that chlor¬amphenicol is markedly different from other antibiotic treatment groups.
Conclusion: Escherichia coli was successfully isolated from cloacal swabs of broiler in Sukabumi, Indonesia. The bacteria were resistant to TE, OT, AMP, CN, NA, CIP, ENR, and erythromycin. Chloramphenicol was more sensitive and effective than other antibiotics in inhibiting the growth of E. coli. The antibiotic resistance genes detected were tetA, blaTEM, gyrA, and ermB.

Key words: Antibiotics; Broiler; Escherichia coli; Gene; Resistance






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