Resistansi Antibiotik Secara Fenotip Dan Deteksi Gen TetA pada Sampel Hati Ayam di Pasar Dukuh Kupang Surabaya

Phenotive Antibiotic Resistance And Detection of TetA Gene in Chicken Liver Samples At Dukuh Market, Kupang Surabaya

Main Article Content

Reina Puspita Rahmaniar
Dyah Widhowati
Nurul Hidayah

Abstract

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.

Downloads

Download data is not yet available.

Article Details

Section
Articles
Author Biographies

Reina Puspita Rahmaniar, Universitas Wijaya Kusuma, Surabaya

Fakultas Kedokteran Hewan Universitas Wijaya Kusuma Surabaya

Dyah Widhowati, Universitas Wijaya Kusuma, Surabaya

Fakultas Kedokteran Hewan, Jl. Dukuh Kupang XXV No. 54, Universitas Wijaya Kusuma Surabaya

Nurul Hidayah, Universitas Wijaya Kusuma, Surabaya

Fakultas Kedokteran Hewan, Jl. Dukuh Kupang XXV No. 54, Universitas Wijaya Kusuma Surabaya

References

Bakhshi M, Fatahi Bafghi M, Astani A, Ranjbar VR, Zandi H, and Vakili M. 2017. Antimicrobial resistance pattern of Escherichia coli isolated from chickens with colibacillosis in Yazd, Iran. Journal of Food Quality and Hazards Control, 4(3), 74–78.
Chuppava B, Keller B, Abd El-Wahab A, Sürie C, and Visscher C. 2019. Resistance Reservoirs and Multi-Drug Resistance of Commensal Escherichia coli From Excreta and Manure Isolated in Broiler Houses With Different Flooring Designs. Frontiers in Microbiology, 10(November), 1–11. https://doi.org/10.3389/fmicb.2019.02633
CLSI. 2018. M100. In The Encyclopedia of Astronomy and Astrophysics. https://doi.org/10.1888/0333750888/6100
Di Francesco A, Salvatore D, Sakhria S, Catelli E, Lupini C, Abbassi MS, Bessoussa G, Ben Yahia S, and Ben Chehida N. 2021. High frequency and diversity of tetracycline resistance genes in the microbiota of broiler chickens in Tunisia. Animals, 11(2), 1–7. https://doi.org/10.3390/ani11020377
Hizlisoy H, Al S, Ertaş Onmaz N, Yildirim Y, Gönülalan Z, and Gümüşsoy KS. 2017. Antimicrobial resistance profiles and virulence factors of Escherichia coli O157 collected from a poultry processing plant. Turkish Journal of Veterinary and Animal Sciences, 41(1), 65–71. https://doi.org/10.3906/vet-1602-22
Ibrahim RA, Cryer TL, Lafi SQ, Basha EA, Good L, and Tarazi YH. 2019. Identification of Escherichia coli from broiler chickens in Jordan, their antimicrobial resistance, gene characterization and the associated risk factors. BMC Veterinary Research, 15(1), 1–16. https://doi.org/10.1186/s12917-019-1901-1
Jahantigh M, Samadi K, Dizaji RE, and Salari S. 2020. Antimicrobial resistance and prevalence of tetracycline resistance genes in Escherichia coli isolated from lesions of colibacillosis in broiler chickens in Sistan, Iran. BMC Veterinary Research, 16(1), 1–6. https://doi.org/10.1186/s12917-020-02488-z
Karami N, Nowrouzian F, Adlerberth I, and Wold AE. 2006. Tetracycline Resistance in Escherichia coli and Persistence in the Infantile Colonic Microbiota. Antimicrobial Agents and Chemotheraphy, 50(1), 156–161. https://doi.org/10.3109/9780203997352.251
Mgaya FX, Matee MI, Muhairwa AP, and Hoza AS. 2021. Occurrence of multidrug resistant escherichia coli in raw meat and cloaca swabs in poultry processed in slaughter slabs in Dar Es Salaam, Tanzania. Antibiotics, 10(4), 0–9. https://doi.org/10.3390/antibiotics10040343
Nguyen F, Starosta AL, Arenz S, Sohmen D, Dönhöfer A, and Wilson DN. 2014. Tetracycline antibiotics and resistance mechanisms. Biological Chemistry, 395(5), 559–575. https://doi.org/10.1515/hsz-2013-0292
Nuraini DM, Andityas M, Paramarta A, Najib NR, and Wijayanti AD. 2020. Isolasi dan identifikasi Escherichia coli dari Sumber Air Minum Kandang Broiler serta Uji Aktivitas Antibakteri Lidah Buaya. Jurnal Ilmu Peternakan Dan Veteriner Tropis (Journal of Tropical Animal and Veterinary Science), 10(2), 106. https://doi.org/10.46549/jipvet.v10i2.116
Olowe OA, Idris OJ, and Taiwo SS. 2013. Prevalence of TET genes mediating tetracycline resistance in Escherichia coli clinical isolates in Osun State, Nigeria . European Journal of Microbiology and Immunology, 3(2), 135–140. https://doi.org/10.1556/eujmi.3.2013.2.7
Putri AR, Suswati E, and Indreswari L. 2018. Resistensi Eschericia coli Dari Isolat Daging Ayam Broiler Terhadap Tetrasiklin Tetracycline Resistant Eschericia coli From Broiler Chicken Meat Isolate 1. Journal of Agromedicine and Medical Sciences, 4(1), 38–44.
Sarker MS, Mannan MS, Ali MY, Bayzid M, Ahad A, and Bupasha ZB. 2019. Antibiotic resistance of Escherichia coli isolated from broilers sold at live bird markets in Chattogram, Bangladesh. Journal of Advanced Veterinary and Animal Research, 6(3), 272–277. https://doi.org/10.5455/javar.2019.f344
Skočková A, Cupáková Š, Karpíšková R, and Janštová B. 2012. Detection of Tetracyline Resistance Genes in Escherichia coli. Lournal of Microbiology, Biotechnology and Food Sciences, 1, 777–784.
Wibisono FJ, Sumiarto B, Untari T, Effendi MH, Permatasari DA, and Witaningrum AM. 2020. Prevalence and Risk Factors Analysis of Multidrug Resistance of Escherichia coli Bacteria in Commercial Chicken, Blitar District. Jurnal Ilmu Peternakan Dan Veteriner Tropis (Journal of Tropical Animal and Veterinary Science), 10(1), 15.
Zhang T, Wang CG, Lv JC, Wang RS, and Zhong XH. 2012. Survey on tetracycline resistance and antibiotic-resistant genotype of avian Escherichia coli in North China. Poultry Science, 91(11), 2774–2777. https://doi.org/10.3382/ps.2012-02453
Zishiri OT, Mkhize N, and Mukaratirwa S. 2016. Prevalence of virulence and antimicrobial resistance genes in Salmonella spp. isolated from commercial chickens and human clinical isolates from south Africa and Brazil. Onderstepoort Journal of Veterinary Research, 83(1), 1–11. https://doi.org/10.4102/ojvr.v83i1.1067