Authors:
Emy Koestanti Sabdoningrum
1
;
Sri Hidanah
1
;
Sri Chusniati
2
;
Wiwik Misaco
3
;
Retno Sri Wahyuni
4
and
Laras Retno Kinasih Harianto
5
Affiliations:
1
Department of Animal Husbandry, Indonesia
;
2
Department of Veterinary Microbiology, Indonesia
;
3
Department of Clinic, Indonesia
;
4
Department of Veterinary Basic Medicine, Indonesia
;
5
Student, Faculty of Veterinary Medicine, Universitas Airlangga and C Campus UNAIR, Indonesia
Keyword(s):
Escherichia coli, Trimethoprim resistance gene, Layer chicken, Polymerase Chain Reaction
Abstract:
Aim: The aim of this research was to detect the presence of a gene which is responsible for Escherichia coli trimethoprim resistance (TEM gene) using Polymerase Chain Reaction from layer chickens in East Java Province, Indonesia. Methods: Purposive sampling of 60 infundibulums has been done in laying chicken farms in those indicating Colibacillosis from 6 districts in East Java Province, Indonesia including Sidoarjo, Mojokerto, Jombang, Kediri, Bojonegoro and Blitar. Ten samples were collected from each district. Samples were isolated using Eosin Methylene Blue Agar and five biochemical tests of Sulfide Indol Motility (SIM), Simons Citrate Agar (SCA), Triple Sugar Iron Agar (TSIA), Urea Agar (UA) and Sugar Test were used for bacteria identification. The DNA was then isolated from Escherichia coli-positive samples prior to Polymerase Chain Reaction (PCR) amplification to investigate TEM gene that indicated trimethoprim antibiotic resistance. Result: Bacteriological tests showed that 3
0 of 60 samples (50%) were E. coli positive. Ten of those were from Blitar, 10 from Bojonegoro, 5 from Sidoarjo, and 5 from Jombang. Two samples from Kediri and Mojokerto districts were negative. From PCR amplification showed that 28 of 30 samples were negative due to TEM gene, and there were 2 positive samples shown from Bojonegoro and Blitar. This indicated that 6.66% of total samples were positive containing TEM gene. Conclusion: Based on bacteriological examination 50% of samples from six districts in East Java, Indonesia were E. coli positive, and 6.66% were trimethoprim resistant based on occurrence of TEM gen in PCR amplification.
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