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Prasanna Vadhana Anbazhagan,1 Prasada Rao Thavitiki,2 Manasa Varra,2 Latchumikanthan Annamalai,3 Ramya Putturu,4 Venkateswara Rao Lakkineni,4 Pavan Kumar Pesingi41Department of Veterinary Microbiology, College of Veterinary Science, Sri Venkateswara Veterinary University, Proddatur, Andhra Pradesh, India; 2Department of Veterinary Biochemistry, College of Veterinary Science, Sri Venkateswara Veterinary University, Proddatur, Andhra Pradesh, India; 3Department of Veterinary Parasitology, Veterinary University Training and Research Centre (VUTRC), TANUVAS, Villupuram, Tamil Nadu, India; 4Department of Veterinary Public Health and Epidemiology, College of Veterinary Science, Sri Venkateswara Veterinary University, Proddatur, Andhra Pradesh, IndiaIntroduction: Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium) is one of the major cause of foodborne zoonoses in humans. Poultry acts as a reservoir for S. Typhimurium without showing clinical signs and has become a source of infection to humans. Besides, it also became a source of multidrug-resistant (MDR) strains of S. Typhimurium.Methods: In the present study, we have isolated 9 S. Typhimurium from 503 samples from environmental sources of poultry wet markets in the Kadapa District of Andhra Pradesh, India. The role of efflux pump activity in antibiotic resistance was evaluated by ethidium bromide cartwheel test and efflux pump inhibition assay.Results: Eight out of nine isolates were resistant to two or more classes of antibiotics. The efflux pump activity of these isolates by ethidium bromide cartwheel method revealed that 66.6% isolates had shown evidence of pronounced efflux activity. The zone of inhibition (ZOI) of resistant antibiotics for each isolate was estimated in the presence or absence of 25 μg/mL of PAβN. Overall, cephalosporins (cefazolin, cefamandole, and cefaclor), kanamycin, polymyxin-B, piperacillin, and imipenem showed significant increase (≥2 mm) of ZOI, indicating the role of efflux pumps for efflux of these drugs. A maximum of 4 antibiotics among EtBr efflux pump positive isolates and 2 antibiotics among EtBr efflux pump negative isolates showed increased ZOI in the presence of PAβN.Conclusion: The results indicate that efflux pumps of MDR S. Typhimurium may contribute to resistance for at least one antibiotic, even in EtBr cartwheel test negative isolates.Keywords: Salmonella, EtBr, efflux, PAβN, ZOI