Deteksi Gen Faktor Virulensi dan Gen Resistansi Siprofloksasin pada Escherichia coli Patogen Asal Ayam Layer

Abstract

Avian Pathogenic Escherichia coli (APEC) merupakan penyebab kolibasilosis pada unggas, Sedangkan E. coli strain O157:H7 merupakan agen zoonosis penyebab penyakit hemolytic uremic syndrome (HUS) pada manusia. Penelitian untuk mengungkap faktor virulensi dan resistansi siprofloksasin pada unggas, khususnya ayam layer di Indonesia, masih sangat jarang. Penelitian ini bertujuan untuk mengetahui gen virulensi dan resistansi siprofloksasin asal APEC serta keberadaan E. coli O157:H7 yang diisolasi dari ayam layer. Siprofloksasin merupakan antibiotik golongan flurokuinolon yang sangat penting pada manusia dan hewan. Siprofloksasin pada manusia masuk kedalam kategori Highest Priority Critically Important Antimicrobials for Human Medicine dan pada hewan masuk kedalam kategori Critically Important Antibiotics for Veterinary Medicine. Penelitian ini menggunakan 327 isolat arsip E. coli Balai Besar Pengujian Mutu dan Sertifikasi Obat Hewan-Kementerian Pertanian Republik Indonesia yang diisolasi dari ayam layer pada tahun 2021 dari tujuh provinsi. Seluruh isolat diuji patogenisitasnya menggunakan Congo Red untuk APEC dan Sorbitol MacConkey agar untuk E. coli O157:H7. E. coli yang dinyatakan patogen diuji resistansinya terhadap siprofloksasin dengan metode agar dilution. Isolat yang resistan dan patogen diuji keberadaan gen virulensi dan resistansinya menggunakan uji polymerase chain reaction (PCR). Hasil uji Congo Red menunjukkan sebanyak 59 isolat E. coli bersifat patogen (APEC), sedangkan E. coli O157:H7 tidak ditemukan. Hasil uji resistansi menunjukkan 30 dari 59 isolat APEC resistan terhadap siprofloksasin. Dari 30 isolat APEC resistan siprofloksasin, ditemukan gen virulensi iss pada 29 isolat (96,7%), iutA pada 23 isolat (76,7%), ompT pada 19 isolat (63,3%), hlyF pada 14 isolat (46,7%), dan iroN pada 11 isolat (36,7%). Gen resistansi gyrA dan parC ditemukan pada 30 isolat (100%), qnrA pada 20 isolat (66,7%), qnrB pada 7 isolat (23%) dan qnrS pada 6 isolat (20%). Penelitian ini menunjukkan bahwa gen virulensi dan resistansi banyak ditemukan pada isolat E. coli patogen (APEC) yang resistan terhadap siprofloksasin. Hal ini menunjukkan adanya ancaman serius terhadap efikasi siprofloksasin untuk mengatasi kolibasilosis pada ayam layer.

Avian Pathogenic Escherichia coli (APEC) is the cause of colibacillosis in poultry while E. coli strain O157:H7 is a zoonotic agent that causes hemolytic uremic syndrome (HUS) disease in humans. Research to reveal virulence factors and ciprofloxacin resistance in poultry of APEC, especially laying hens in Indonesia is still very rare. This study aimed to determine virulence and ciprofloxacin resistance genes from APEC and E. coli strain O157:H7 isolated from laying hens. Ciprofloxacin is a fluoroquinolone antibiotic which is very important in humans and animals. Ciprofloxacin in humans is included in the category of Highest Priority Critically Important Antimicrobials for Human Medicine and in animals it is included in the category of Critically Important Antibiotics for Veterinary Medicine.This research used 327 E. coli isolates from the archives of the National Veterinary Drug Assay Laboratory-Ministry of Agriculture, which were isolated from laying hens in 2021 from seven provinces. All isolates were tested for pathogenicity using Congo Red for APEC and Sorbitol MacConkey agar for E. coli O157:H7. E. coli which was declared pathogenic was tested for resistance to ciprofloxacin using agar dilution method. Resistant and pathogenic isolates are tested for the presence of virulence and resistance genes using polymerase chain reaction test. The results of the Congo Red test, it was found that 59 were APEC while E. coli O157:H7 was not found. The resistance test results; 30 of 59 pathogenic E. coli isolates were ciprofloxacin-resistant. From 30 pathogenic E. coli ciprofloxacin-resistant isolates, the virulence gene iss was found in 29 isolates (96,7%), iutA in 23 isolates (76.7%), ompT in 19 isolates (63.3%), hlyF in 14 isolates (46.7 %) and iroN in 11 isolates (36.7%). The resistance genes gyrA and parC were found in 30 isolates (100%), qnrA in 20 isolates (66.7%), qnrB in 7 isolates (23%) and qnrS in 6 isolates (20%). This research shows that virulence and resistance genes are often found in pathogenic E. coli ciprofloxacin-resistant isolates. This indicates a serious threat to the efficacy of ciprofloxacin to overcome colibacillosis in laying hens.