Phenotypic and genotypic examination of antimicrobial resistance in thermophilic Campylobacter species isolated from poultry in Turkey


Introduction: The study aimed to isolate thermophilic Campylobacter from chickens raised three rearing methods, determine its antimicrobial susceptibilities, and examine resistance-related genes by PCR.

Material and Methods: Cloacal swabs or intestinal contents were taken in Istanbul, Sakarya, and Izmir provinces. Chickens were from small village-based family-run businesses (n = 70), organically raised (n = 71), and conventionally raised broilers (n = 79). The samples were cultured on modified charcoal cefoperazone desoxycholate (mCCD) agar. Suspect isolates were identified with multiplex PCR (mPCR). As per EUCAST standards, MIC values were derived by broth microdilution for tetracycline, ciprofloxacin, nalidixic acid, kanamycin, gentamicin, and erythromycin in isolates of C. jejuni (n = 98) and C. coli (n = 83).

Results: In C. jejuni, 78.6% tetracycline, 87.8% ciprofloxacin, and 81.6% nalidixic acid resistance was detected, but none was to kanamycin, gentamicin, or erythromycin. In C. coli, 98.8% ciprofloxacin and 63.9% nalidixic acid resistance was detected, whereas resistance to nonquinolones was not observed. C257T (Thr-86-Ile) mutation in the gyrA gene of all phenotypically quinolone-resistant isolates was detected through a mismatch amplification mutation assay PCR (MAMA-PCR). It emerged that all isolates bore the tet (O) resistance gene.

Conclusion: Common tetracycline, nalidixic acid, and ciprofloxacin resistance exists in Campylobacter isolated from chickens raised three rearing methods.

If the inline PDF is not rendering correctly, you can download the PDF file here.

  • 1. Abay S., Kayman T., Otlu B., Hizlisoy H., Aydin F., Ertas N.: Genetic diversity and antibiotic resistance profiles of Campylobacter jejuni isolates from poultry and humans in Turkey. Int J Food Microbiol 2014, 178, 29–38.

  • 2. Alfredson D.A., Korolik V.: Antibiotic resistance and resistance mechanisms in Campylobacter jejuni and Campylobacter coli. FEMS Microbiol Lett 2007, 277, 123–132.

  • 3. Alonso R., Mateo E., Churruca E., Martinez I., Girbau C., Fernández-Astorga A.: MAMA-PCR assay for the detection of point mutations associated with high-level erythromycin resistance in Campylobacter jejuni and Campylobacter coli strains. J Microbiol Methods 2005, 63, 99–103.

  • 4. Aslantaş Ö.: Genotypic, antimicrobial resistance and virulence profiles of thermophilic Campylobacter isolates in broilers. Kafkas Univ Vet Fak Derg 2017, 23, 547–554.

  • 5. Chen X., Naren G.W., Wu C.M., Wang Y., Dai L., Xia L.N., Luo P.J., Zhang Q., Shen J.Z.: Prevalence and antimicrobial resistance of Campylobacter isolates in broilers from China. Vet Microbiol 2010, 144, 133–139.

  • 6. Cokal Y., Caner V., Sen A., Cetin C., Karagenc N.: Campylobacter spp. and their antimicrobial resistance patterns in poultry: An epidemiological survey study in Turkey. Zoonoses Public Health 2009, 56, 105–110.

  • 7. Corry J.E.L., Atabay H.I.: Handbook of culture media for food and water microbiology. In: Culture Media for the Isolation of Campylobacters, Helicobacters, and Arcobacters, edited by Curtis G.D.W. and Baird R.M., Royal Society of Chemistry, Cambridge, 2012, pp. 403–435.

  • 8. European Food Safety Authority (EFSA).: Trends and sources of zoonoses and zoonotic agents and food-borne outbreaks in 2011. EFSA J 2013, 11, 1–250.

  • 9. El-Adawy H., Hotzel H., Düpre S., Tomaso H., Neubauer H., Hafez H.M.: Determination of antimicrobial sensitivities of Campylobacter jejuni isolated from commercial turkey farms in Germany. Avian Dis 2012, 56, 685–692.

  • 10. El-Adawy H., Ahmed M.F., Hotzel H., Tomaso H., Tenhagen B.A., Hartung J., Neubauer H., Hafez H.M.: Antimicrobial susceptibilities of Campylobacter jejuni and Campylobacter coli recovered from organic turkey farms in Germany. Poult Sci 2015, 94, 2831–2837.

  • 11. Englen M.D., Hill A.E., Dargatz D.A., Ladely S.R., Fedorka‐Cray P.J.: Prevalence and antimicrobial resistance of Campylobacter in US dairy cattle. J Appl Microbiol 2007, 102, 1570–1577.

  • 12. Ertaş H., Çetinkaya B., Muz A., Öngör H.: Identification of chicken originated Campylobacter coli and Campylobacter jejuni by polymerase chain reaction (PCR). Turk J Vet Anim Sci 2014, 26, 1447–1452.

  • 13. European Committee on Antimicrobial Susceptibility Testing (EUCAST).: Preparation of nutrients for determination of MIC values by disk diffusion test and liquid microdilution method. EUCAST 2014, 4, 1–5.

  • 14. Fallon R., O’Sullivan N., Maher M., Carroll C.: Antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolates from broiler chickens isolated at an Irish poultry processing plant. Lett Appl Microbiol 2003, 36, 277–281.

  • 15. Gibreel A., Sköld O., Taylor D.E.: Characterization of plasmid-mediated aphA-3 kanamycin resistance in Campylobacter jejuni. Microb Drug Resist 2004, 10, 98–105.

  • 16. Hariharan H., Sharma S., Chikweto A., Matthew V., DeAllie C.: Antimicrobial drug resistance as determined by the E-test in Campylobacter jejuni, C. coli, and C. lari isolates from the ceca of broiler and layer chickens in Grenada. Comp Immunol Microbiol Infect Dis 2009, 32, 21–28.

  • 17. Hizlisoy H., Kilic H.: Broiler Karkaslarından İzole Edilen Campylobacter jejuni İzolatlarının Makrolid, Kinolon ve Tetrasiklin Grubu Antibiyotiklere Karşı Direnç Durumu. (The resistance to macrolide, quinolone, and tetracycline group antibiotics of campylobacter jejuni isolates isolated from broiler carcasses) Erciyes Üniv Vet Fak Derg 2015, 12, 81–92.

  • 18. Hungaro H.M., Mendonça R.C.S., Rosa V.O., Badaró A.C.L., Moreira M.A.S., Chaves J.B.P.: Low contamination of Campylobacter spp. on chicken carcasses in Minas Gerais state, Brazil: molecular characterization and antimicrobial resistance. Food Control 2015, 51, 15–22.

  • 19. International Organization for Standardization (ISO). Clinical laboratory testing and in vitro diagnostic test systems-susceptibility testing of infectious agents and evaluation of performance of antimicrobial susceptibility test devices-Part 1. ISO 2006, 20776, 1–19.

  • 20. Josefsen M.H., Lübeck P.S., Hansen F., Hoorfar J.: Towards an international standard for PCR-based detection of foodborne thermotolerant Campylobacters: interaction of enrichment media and pre-PCR treatment on carcass rinse samples. J Microbiol Methods 2004, 58, 39–48.

  • 21. Kurinčič M., Klančnik A., Smole Možina S.: Effects of efflux pump inhibitors on erythromycin, ciprofloxacin, and tetracycline resistance in Campylobacter spp. isolates. Microb Drug Resist 2012, 18, 492–501.

  • 22. Kurekci C., Önen S.P.: Characteristics of ciprofloxacin resistant Campylobacter spp. isolated from chicken meat in Turkey. J Food Meas Charact 2017, 11, 586–591.

  • 23. Linton D., Owen R.J., Stanley J.: Rapid identification by PCR of the genus Campylobacter and of five Campylobacter species enteropathogenic for man and animals. Res Microbiol 1996, 147, 707–718.

  • 24. Luangtongkum T., Morishita T.Y., Ison A.J., Huang S., McDermott P.F., Zhang Q.: Effect of conventional and organic production practices on the prevalence and antimicrobial resistance of Campylobacter spp. in poultry. Appl Environ Microbiol 2006, 72, 3600–3607.

  • 25. Maćkiw E., Korsak D., Rzewuska K., Tomczuk K., Rożynek E.: Antibiotic resistance in Campylobacter jejuni and Campylobacter coli isolated from food in Poland. Food Control 2012, 23, 297–301.

  • 26. Müller W., Böhland C., Methner U.: Detection and genotypic differentiation of Campylobacter jejuni and Campylobacter coli strains from laying hens by multiplex PCR and fla-typing. Res Vet Sci 2011, 91, 48–52.

  • 27. Olah P.A., Doetkott C., Fakhr M.K., Logue C.M.: Prevalence of the Campylobacter multi-drug efflux pump (CmeABC) in Campylobacter spp. isolated from freshly processed turkeys. Food Microbiol 2006, 23, 453–460.

  • 28. Ozdamar, K.: Biostatistics with SPSS. Kaan Bookstore, Eskisehir, 1999.

  • 29. Sahin O., Kassem I.I., Shen Z., Lin J., Rajashekara G., Zhang Q.: Campylobacter in poultry: ecology and potential interventions. Avian Dis 2015, 59, 185–200.

  • 30. Savaşan S., Çiftçi A., Diker K.S.: Emergence of quinolone resistance among chicken isolates of Campylobacter in Turkey. Turk J Vet Anim Sci 2004, 28, 391–397.

  • 31. Shin E., Lee Y.: Antimicrobial resistance of 114 porcine isolates of Campylobacter coli. Int J Food Microbiol 2007, 118, 223–227.

  • 32. Yildirim M., Istanbulluoğlu E., Ayvali B.: Prevalence and antibiotic susceptibility of thermophilic Campylobacter species in broiler chickens. Turk J Vet Anim Sci 2005, 29, 655–660.

  • 33. Zhao S., Young S.R., Tong E., Abbott J.W., Womack N., Friedman S.L., McDermott P.F.: Antimicrobial resistance of Campylobacter isolates from retail meat in the United States between 2002 and 2007. Appl Environ Microbiol 2010, 76, 7949–7956.

  • 34. Zirnstein G., Helsel L., Li Y., Swaminathan B., Besser J.: Characterization of gyrA mutations associated with fluoroquinolone resistance in Campylobacter coli by DNA sequence analysis and MAMA PCR. FEMS Microbiol Lett 2000, 190, 1–7.

  • 35. Zirnstein G., Li Y., Swaminathan B., Angulo F.: Ciprofloxacin resistance in Campylobacter jejuni isolates: detection of gyrA resistance mutations by mismatch amplification mutation assay PCR and DNA sequence analysis. J Clin Microbiol 1999, 37, 3276–3280.

Journal + Issues