Detection of Ciprofloxacin resistance genes in Escherichia coli isolated from dogs with urinary tract infections

Document Type : Research Articles


1 Department of Veterinary Medicine, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana India.

2 Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana India.


This research was performed on uropathogenic Escherichia coli (E. coli) isolates and established the genes of resistance to ciprofloxacin between the isolates. A total of one hundred and three urine samples were tested for uropathogenic E. coli which were obtained from dogs with urinary tract infections (UTIs) using cultural isolation, antimicrobial susceptibility test, and polymerase chain reaction (PCR). The results revealed that genes associated with ciprofloxacin resistance are 24.3% positive for E. coli. The E. coli isolates were resistant to both ciprofloxacin and ampicillin (100%), highly susceptible to chloramphenicol (84.0%), and less susceptible to gentamycin (44.0%) and amikacin (40.0%). The PCR tests showed the presence of the ParC (in 25 samples; 100%), GyrA (in 25 samples; 100%), and GyrB (in 4 samples; 16.0%) genes. The findings of the present study showed an upsetting rate of ciprofloxacin and ampicillin resistance among the E. coli isolates from dogs with UTIs.


Main Subjects

. Chen YM, Wright PJ, Lee CS, Browning GF. Uropathogenic virulence factors in isolates of Escherichia coli from clinical cases of canine pyometra and feces of healthy bitches. Vet. Microbiol. 2003; 94: 57–69.
2. Shaheen BW, Boothe DM, Oyarzabal OA, Wang C, Johnson CM. Evaluation of the contribution of gyrA mutation and efflux pumps to fluoroquinolone and multidrug resistance in pathogenic Escherichia coli isolates from dogs and cats. Am. J. Vet. Res. 2011a; 72: 25–32.
3. Shaheen BW, Nayak R, Foley SL, Kweon O, Deck J, Park M, Rafii F, Boothe DM. Molecular characterization of resistance to extended-spectrum cephalosporins in clinical Escherichia coli isolates from companion animals in the United States. Antimicrob. Agents Chemother. 2011b; 55: 5666–5675.
4. Gibson JS, Cobbold RN, Kyaw-Tanner MT, Heisig P, Trott DJ. Fluoroquinolone resistance mechanisms in multidrug-resistant Escherichia coli isolated from extraintestinal infections in dogs. Vet. Microbiol. 2010; 146: 161–166.
5. Cooke CL, Singer RS, Jang SS, Hirsh DC. Enrofloxacin resistance in Escherichia coli isolated from dogs with urinary tract infections. J. Am. Vet. Med. Assoc. 2002; 220: 190–192.
6. Sanchez S, McCrackin SMA, Hudson CR, Maier M, Buffington T, Dam Q, Maurer JJ. Characterization of multidrug resistant Escherichia coli isolates associated with nosocomial infections in dogs. J. Clin. Microbiol. 2002; 40: 3586–3595.
7. Liu X, Boothe DM, Thungrat K, Aly S. Mechanisms accounting for fluoroquinolone multidrug resistance Escherichia coli isolated from companion animals. Vet. Microbiol. 2012; 161: 159–168.
8. Wetzstein HG. Comparative mutant prevention concentrations of pradofloxacin and other veterinary fluoroquinolones indicate differing potentials in preventing selection of resistance. Antimicrob. Agents Chemother. 2005; 49: 4166–4173.
9. Pan XS, Ambler J, Mehtar S, Fisher LM. Involvement of topoisomerase IV and DNA gyrase as ciprofloxacin targets in Streptococcus pneumoniae. Antimicrob. Agents Chemother. 1996; 40: 2321–2326.
10. Pestova E, Millichap JJ, Noskin GA, Peterson LR. Intracellular targets of moxifloxacin: a comparison with other fluoroquinolones. J. Antimicrob. Chemother. 2000; 45: 583–590.
11. Cohn LA, Gary AT, Fales WH, Madsen RW. Trends in fluoroquinolone resistance of bacteria isolated from canine urinary tracts. J. Vet. Diagn Invest. 2003, 15:338–343.
12. Bauer AW, Kirby WMM, Sherris JC, Turck M. Antibiotic susceptibility testing by standardized single disk method. Ame. J. Clin. Patho. 1966; 45: 493-496.
13. Yoshida H, Bogaki M, Nakamura M, Nakamura S. Quinolone resistance-determining region in the DNA gyrase gyrA gene of Escherichia coli. Antimicrob. Agnts. Chemother. 1990; 34: 1271-1272.
14. Peng H, Marians KJ. Escherichia coli topoisomerase IV, Purification, characterization, subunit structure and subunit interactions J. Biol.Chem. 1993; 268: 24481-24490.
15. Adachi T, Mizuuchi M, Robinson EA, Appella E, O’Dea MH, Gellert M. Mizuuchi K. DNA sequence of the E. coli gyrB gene: application of a new sequencing strategy, Nucl. Aci. Res. 1987; 15: 771-784.
16. Johnson JR. Microbial virulence determinants and the pathogenesis of urinary tract infection. Infect. Dis. Clin. North. Am. 2003; 17(2): 261–78.
17. Brolund A. Overview of ESBL producing Enterobacteriaceae from a Nordic perspective Infec. Ecol. Epidemiol. 2014; 4(1): 1-9.
18. Chang SK, Lo DY, Wei HW, Kuo HC. Antimicrobial resistance of Escherichia coli isolates from canine urinary tract infections. J. Vet. Med. Sci. 2015; 77 (1): 59-65.
19. Moyaert H, Morrissey I, de Jong A, El Garch F, Klein U, Ludwig C, Thiry J, Youala M. Antimicrobial susceptibility monitoring of bacterial pathogens isolated from urinary tract infections in dogs and cats across Europe: ComPath Results. Microbial.Drug Resist.2016; 23(3):391-403.
20. Kuan NL, Chang CH, Lee CA, Yeh KS. Extended spectrum beta lactamaseproducing Escherichia coli and Klebsiella pneumonia isolates from the urine of dogs and cats suspected of urinary tract infection in a veterinary teaching hospital. Tawn. Vet. J. 2016; 42 (3): 143 148.
21. Liu X, Liu H, Li Y, Hao C. Association between virulence profile and fluoroquinolone resistance in Escherichia coli isolated from dogs and cats in China. Infect. Dev. Ctries. 2017; 11(4): 306-313.
22. Ruiz J. Mechanism of resistance to quinolones: Target alterations, decreased accumulation and DNA gyrase protection. J. Antimicrob. Chemother. 2003; 51: 1109-1117.
23. Literak I, Reitschmied T, Bujnakova D, Dolejka M, Cizek A, Bardon J, Pokludova L, Alexa P, Halova D, Jamborova I. Broilers as a source of quinolone-resistant and extraintestinal pathogenic Escherichia coli in the Czech Republic. Micob. Drug. Resist. 2013; 19: 57-63.
24. Oliveira M, Dias FR, Pomba C. Biofilm and fluoroquinolone resistance of canine Escherichia coli uropathogenic isolates. BMC Res. Not. 2014; 7: 499.
25. Thungrat K, Price SB, Carpenter DM, Boothe DM. Antimicrobial susceptibility patterns of clinical Escherichia coli isolates from dogs and cats in the United State: January 2008 through January 2013. Vet. Microbiol. 2015; 179: 287-295.
26. Hebru E, Choi MJ, Lee SJ, Damte D, Park SC. Mutant-prevention concentration and mechanism of resistance in clinical isolates and enrofloxacin/marbofloxacin selected mutants of Escherichia coli of canine origin. J. Med. Microbiol. 2011; 60: 1512-1522.
27. Krishnan S, Balasubramania D, Raju BA, Lakshmi BS. Use of naturally occurring codon bias for identifying topoisomerase mutations in ciprofloxacin resistant Escherichia coli using PCR and future prospects with other bacterial genera: A pilot study. Adv. Biol. Chem. 2012; 2: 366-371.
28. Sunde M, Sorum H. Characterization of integrons in Escherichia coli of the normal intestinal flora of swine. Microb. Drg. Resist. 1999; 5: 279-287.
29. Livermore DM. Β Lactamases in laboratory and clinical resistance. Clin. Microbiol. Rev. 1995; 8: 557-584.
30. Lei T, Tian W, He L, Huang X.H, Sun YX, Deng YT, Sun Y, Lv DH, Wu CM, Huang LZ, Shen, JZ, Liu JH. Antimicrobial resistance in Escherichia coli isolates from food animals, animal food products and companion animals in China. Vet. Microbiol. 2010; 146: 85-89.
31. Gilliver M, Bennett M, Begon M, Hazel S, Hart C. Enterobacteria: antibiotic resistance found in wild rodents. Natr. 1999; 401: 233-234.
32. Wedley AL, Maddox TW, Westgarth C, Coyne KP, Pinchbeck GL, Williams NJ, Dawson S. Prevalence of antimicrobial-resistant Escherichia coli in dogs in a cross-sectional community-based study. Vet. Rec. 2011; 168(13): 354.
33. Wong C, Epstein SE, Westropp JL. Antimicrobial susceptibility patterns in urinary tract infections in dogs (2010–2013). Vet. Intern. Med. 2015; 29: 1045–1052.
34. Nhung NT, Cuong NV, Campbell J, Hoa NT, Bryan JE, Truc VNT, Kiet BT, Jombart T, Trung NV, Hien VB, Thwaites G, Baker S, Carrique-Mas J. High levels of antimicrobial resistance among Escherichia coli isolates from livestock farms and synanthropic rats and shrews in the Mekong Delta of Vietnam. App. Environ. Microbiol. 2015; 81: 812-820.
35. Liu C, Zheng H, Yang M, Xu Z, Wang X, Wei L, Tang B, Liu F, Zhang Y, Din Y., Tang X, Wu B, TJ, Johnson TJ, Chen H, Tan C. Genome analysis and in vivo virulence of porcine extraintestinal pathogenic Escherichia coli strain PCN033. BMC. Genom. 2015; 16(717): 1-18.
36. Cavalho AC, Barbosa AV, Arais LR, Ribeiro PF, Carneiro VC, Cerqueira AMF. Resistance patterns, ESBL gene, and genetic relatedness of Escherichia coli from dogs and owners. Brazil. J. Microbiol. 2016; 47: 150-158.
37. Allen SE, Boerlin P, Janecko N, Lumsden JS, Barker IK, Pearl DL, Reid-Smith RJ, Jardine, C. Antimicrobial resistance in generic Escherichia coli isolates from will small mammals living in swine farm, residential, landfill and natural environments in southern Ontario, Canada. Appl. Environ. Microbiol. 2011; 77: 882-888.
38. Mustapha M, Parveen G, Vinay K, Divya A, Tarun K, Sushila M. Isolation of uropathogenic Escherichia coli from dogs and molecular detection of chloramphenicol resistance genes. Haryana Vet. 2019; 58, 66-69.
39. Dechet AM, Scallan E, Gensheimer K, Hoekstra R, Gunderman-King J, Lockett J, Wrigley D, Chege W, Sobel J, Multistate Working Group. Outbreak of multidrug resistant Salmonella enteric serotype Typhimurium Definitive Type 104 infection linked to Commercial ground beef, northeastern United States, 2003-2004. Clin. Infect. Dis. 2006; 42(6): 747-752.
40. Cummings KJ, Warnick LD, Davis MA, Eckmann K, Gröhn YT, Hoelzer K, MacDonald K, Root TP, Siler JD, McGuire SM, Wiedmann M, Wright EM, Zansky SM, Besser TE. Farm animal contact as risk factor for transmission of bovine associated Salmonella subtypes. Emerg. Infect. Dis. 2012; 18: 1929–1936.
41. Simjee S, White DG, McDermott PF, Wanger DD, Zervos MJ, Donabedia SM, English LL, Hayes JR, Walker RD. Characterization of Tn1546 in vancomycin resistant Enterococcus faecium isolated from canine urinary tract infections: Evidence of gene exchange between human and animal enterococci. J. Clin. Microbiol. 2002; 40: 4659 – 4665.
42. Stockholm J, Schjorring S, Pedersen L, Bischoff A, Følsgaard N, Carson CG, Chawes B, Bønnelykke K, Mølgaard A, Krogfelt KA, Bisgaard H. Living with cat and dog increases vaginal colonization with E. coli in pregnant women. PLoS One. 2012; 7(9): 1-6.
43. Onanuga A, Oyi AR, Onaolapa AJ. Prevalence and susceptibility pattern of Methicillin resistant Staphylococcus aureus isolates amongst healthy women in Zaria, Nigeria. Afri. J. Biotech. 2005; 4(11): 1321-1324.
44. Poole K. Efflux mediated resistance to fluoroquinolones in gram negative bacteria. Antimicrob. Agnts. Chemother. 2000; 44: 2233-2241.