190 P-ISSN: 2304-3075; E-ISSN: 2305-4360 International Journal of Veterinary Science www.ijvets.com; editor@ijvets.com Research Article Detection of Quinolone Resistance through Amplification of the gyrA Gene of Mycobacterium species from Human and Animal Sources Gemerlyn G Garcia 1,§ , Ralph Kevin M Espinosa 1 , Michelle A Miguel 2 , Mark Lester Bernardino 3 ,Mark Arman D Aquino 1 and Claro N Mingala 2,§ * 1 College of Veterinary Science and Medicine, Central Luzon State University; 2 Biosafety and Environment Section, Philippine Carabao Center National Headquarters and Gene Pool, Science City of Munoz 3120, Nueva Ecija, Philippines; 3 Tropical Disease Foundation, Inc., Philippine Institute of Tuberculosis Building, Amorsolo St. corner Urban Ave., Bgy. Pio Del Pilar, Makati City 1230, Metro Manila, Philippines § Both authors contributed equally to this work *Corresponding author: cnmingala@hotmail.com Article History: Received: December 13, 2017 Revised: September 09, 2018 Accepted: October 12, 2018 ABSTRACT Ten (10) DNA samples of Mycobacterium tuberculosis (Mtb) isolated from sputum of TB-positive humans, DNA samples from Mycobacterium species isolated from lymph nodes and fecal samples of avians and bubaline animals were analysed by PCR targeting primers for gyrase A (gyrA), quinolone resitance A(qnrA) and topoisomerase IV (parC) genes. Results demonstrated that quinolone resistance recognized by gyrA was seen in one out of 10 DNA samples from human Mtb isolates and that no qnrA and parC genes were detected. The gene for quinolone resistance detected by the primer gyrA had a molecular weight of 333 bp. Resistance to quinolone mediated by gyrA, qnrA and parC genes in avian (M. avium avium) and bubaline (M. avium paratuberculosis) isolates of mycobacteria were not detected after PCR. The non-amplification of genes observed in this study explains the non-existence of quinolone resistance arbitrated by gyrA, qnrA and parC genes in the specified avian and bubaline mycobacterial isolates. Key words: Mycobacterium avium avium, Mycobacterium avium paratuberculosis, Mycobacterium tuberculosis , Polymerase chain reaction, Quinolone resistance INTRODUCTION Fluroquinolones are synthetic antibiotics that inhibit nucleic acid synthesis in bacteria. Original preparations of quinolones reportedly act against enteric bacteria and some facultative Gram-negative microbes. The fluorinated quinolones, now known as fluoroquinolones, were purportedly made from original preparations to augment the spectrum of activity against other bacteria like Pseudomonas, Gram-positive and Gram-negative bacteria. Reduced sensitivity of bacteria to the original classes of quinolones reportedly observed by many researchers parallel build up of resistance to the drug (Fung-Tome et al., 1993). Currently available preparations of fluoroquinolones used in human medicine include moxifloxacin, gatifloxacin, levofloxacin, ofloxacin and ciprofloxacin (Ginsburg et al., 2003; Huang and Stafford, 2002; Wallace et al., 1990). In veterinary practice, fluoroquinolones such as enrofloxacin, levofloxacin and ciprofloxacin are used as vital medications for many systemic infections of livestock, poultry and fish in many parts of the world. The use of these drugs in livestock presents an area of concern due to the possible transmission of antimicrobial resistance to humans via the food chain (Endtz et al., 1991). Bacteria that develop resistance to quinolones generally develop cross- resistance to other quinolones and this cross-resistance reportedly covers quinolones used in animals and humans. Bacterial exposure to lower concentrations of quinolones increases build-up of resistance (Hooper, 2011). Inadequate dosing of quinolones, different routes of administration and treatment of water and feed supplementation with quinolones are common controversies that underlie induction of resistance (Herikstad et al., 1997). With the introduction of advanced methods in molecular biology, various mechanisms such as chromosomal mutation (Takiffet al., 1994), plasmid-intervention (Tran et al., 2009) and alteration of DNA gyrase (Cambau et al., 1994) have been identified in the acquisition of quinoloneresistance by bacteria. Cite This Article as: GG Garcia, RKM Espinosa, MA Miguel, ML Bernardino,MAD Aquino and CN Mingala, 2018. Detection of quinolone resistance through amplification of the gyra gene of mycobacterium species from human and animal sources. Inter J Vet Sci, 7(4): 190-194. www.ijvets.com (©2018 IJVS. All rights reserved)