Resistansi Antibiotik Secara Fenotip Dan Deteksi Gen TetA pada Sampel Hati Ayam di Pasar Dukuh Kupang Surabaya
Authors: Rahmaniar Reina Puspita , Widhowati Dyah , Hidayah Nurul 
Published: January 2022

Abstract

The purpose of this study was to determine the resistance of several antibiotics phenotypically and genotypically to detect the tetA gene from broiler chicken liver samples at Dukuh Kupang market, Surabaya. A total of 30 samples were taken and then prepared aseptically and sterile. Isolation on Eosin Methylene Blue Agar (EMBA) media, then microscopic examination using gram staining and biochemical tests of Triple Sugar Iron Agar (TSIA), Sulfide Indole Motility (SIM), Methyl Red (MR), Voges Prouskauers (VP) and Simons Citrate Agar (SCA). The identified Escherichia coli colonies were tested for antibiotic sensitivity using the Kirby Bauer method, then isolates that were proven to be resistant to tetracycline antibiotics were followed by genetic testing using the Polymerase Chain Reaction (PCR) method. The results showed that 20 of the 30 samples were positive for Escherichia coli. Escherichia coli isolates from chicken liver samples showed resistance to 30 µg tetracycline antibiotics by 85% (17 of 20 samples) Researchers also compared with other antibiotics, the highest resistance to ampicillin 10 µg was 90% (18 out of 20 samples), gentamicin resistance was 10 µg by 50% (10 of 20 samples) and 30 µg chloramphenicol antibiotic resistance by 30% (6 of 20 samples). The isolates that were resistant to tetracycline were confirmed by Polymerase Chain Reaction to detect the tetA gene with the final product in the form of a band with a length of 210 bp. Bacterial isolates resistant to Tetracycline antibiotics did not always show TetA gene expression in the PCR test.

Keywords: Antibiotic Resistance; Escherichia coli; Market; TetA gene

 

Abstrak

Tujuan dari penelitian ini yaitu untuk mengetahui resistansi beberapa antibiotik secara fenotip dan secara genotip mendeteksi gen tetA dari sampel hati ayam broiler di pasar Dukuh Kupang Surabaya. Sebanyak 30 sampel diambil kemudian dipreparasi secara aseptis dan steril. Isolasi pada media Eosin Methilen Blue Agar (EMBA), selanjutnya dilakukan pemeriksaan mikroskopis menggunakan pewarnaan gram dan uji biokimiawi Triple Sugar Iron Agar (TSIA), Sulfide Indol Motility (SIM), Methyl Red (MR), Voges Prouskauers (VP), dan Simons Citrat Agar (SCA). Koloni Escherichia coli yang teridentifikasi dilakukan uji sensitifitas antibiotik dengan metode Kirby bauer, selanjutnya isolat yang terbukti resistan terhadap antibiotik tetrasiklin dilanjutkan pemeriksaan genetik dengan metode Polymerase Chain Reaction (PCR).  Hasil penelitian menunjukkan bahwa 20 dari 30 sampel positif Escherichia coli. Isolat Escherichia coli asal sampel hati ayam menunjukkan resistansi terhadap antibiotik Tetrasiklin 30 µg sebesar 85 % (17 dari 20 sampel) Peneliti juga melakukan perbandingan dengan antibiotik lainnya, resistensi tertinggi pada antibiotik ampisilin 10 µg sebesar 90 % (18 dari 20 sampel), resistensi gentamisin 10 µg sebesar 50 % (10 dari 20 sampel) dan resistensi antibiotik kloramfenikol 30 µg sebesar 30 % (6 dari 20 sampel). Isolat yang resisten terhadap tetrasiklin dikonfirmasi dengan Polymerase Chain Reaction untuk mendeteksi gen tetA dengan produk akhir berupa band dengan panjang 210 bp. Isolat bakteri yang resistan terhadap antibiotik Tetrasiklin tidak selalu menunjukkan ekspresi gen tetA pada uji PCR.

Kata kunci: Escherichia coli; Gen TetA; Pasar; Resistansi Antibiotik.