Isolation and Antibiotic Resistance of Helicobacter pullorum from Chicken Wings

Document Type : Research Paper


1 DVM Graduated, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran.

2 Department of Clinical Science, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran.

3 Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran.


Introduction: Helicobacter pullorum has recently emerged as a significant foodborne pathogen. The present study aimed to isolate and determine the antibiotic resistance of H. pullorum from fresh chicken wing using the culture method and a molecular technique in Semnan, Iran. Methods: A total of 60 fresh chicken wings samples were purchased from various local retail markets in different regions of Semnan and processed using the culture method. For initial confirmation, biochemical tests were applied. Suspected colonies were subjected to polymerase chain reaction (PCR) by the 16S rRNA gene. The antibiotic resistance of the isolates was also assessed using the disk-diffusion method. Results: Among 60 samples, 27 (45%) were H. pullorum-positive based on the culture method and biochemical tests. However, the PCR test indicated 18 samples (30%) to be positive for H. pullorum. In the antibiogram, the highest and lowest resistance rates were observed against ciprofloxacin and fosfomycin, respectively. Conclusion: This was the first report in Iran to clearly illustrate that H. pullorum could be found in fresh chicken wings at a moderate level. In addition, the antibiotic resistance of the H. pullorum isolates was confirmed, and the PCR test based on the 16S rRNA gene was considered a reliable and sensitive technique for the detection of this pathogen. However, further investigation is required to explore the life cycle of this novel foodborne pathogen in the other regions of Iran.


  1. Jebelli Javan A, Emadi Chashmi S H, Staji H, Akhlaghi, H. Comparison of the culture and PCR methods to determine the prevalence and antibiotic resistance of Helicobacter pullorum isolated from chicken thigh samples in Semnan, Iran. J Human Environ Health Promot. 2020; 6(4): 167-72.
  2. Jebelli Javan A, Emadi Chashmi S H, Staji H, Akhlaghi, H. Comparison of culture and PCR method in prevalence of Helicobacter pullorum isolated from chicken liver samples and determination of antibiotic resistance profile of the isolates. J Vet Microbiol. 2020; 16(2): 73-83.
  3. Suzuki H, Yamamoto S. Campylobacter contamination in retail poultry meats and by-products in Japan: A literature survey. Food Control. 2009; 20(6): 531-7.
  4. Akhlaghi H, Emadi Chashmi S H, Jebelli Javan A. Prevalence and Antibiotic Resistance of Helicobacter pullorum isolates in poultry from Semnan province, Iran. Int J Enteric Pathog. 2020; 8(3): 101-6.
  5. Borges V, Santos A, Correia C B, Saraiva M, Ménard A, Vieira L, et al. Helicobacter pullorum isolated from fresh chicken meat: Antibiotic resistance and genomic traits of an emerging foodborne pathogen. Appl Environ Microbiol. 2015; 81(23): 8155-63.
  6. Qumar S, Majid M, Kumar N, Tiwari S K, Semmler T, Devi S, et al. Genome dynamics and molecular infection epidemiology of multidrug-resistant Helicobacter pullorum isolates obtained from broiler and free-range chickens in India. Appl Environ Microbiol. 2017; 83(1): e02305-16.
  7. Akhlaghi H, Emadi Chashmi S H, Jebelli Javan A. Development of a novel and specialized cultivation method for isolating Helicobacter pullorum from chicken meat. Iran J Vet Res. 2021; 22(1): 76-80.
  8. Zanoni R G, Rossi M, Giacomucci D, Sanguinetti V, Manfreda G. Occurrence and antibiotic susceptibility of Helicobacter pullorum from broiler chickens and commercial laying hens in Italy. Int J Food Microbiol. 2007; 116(1): 168-73.
  9. Ochoa S, Martínez O A, Fernández H, Collado L. Comparison of media and growth conditions for culturing enterohepatic Helicobacter species. Lett Appl Microbiol. 2019; 69(3): 190-7.
  10. Stanley J, Linton D, Burnes A P, Dewhirt F E, On S L W, Porter A, et al. Helicobacter pullorum sp. nov. genotype and phenotype of a new species isolated from poultry and from human patients with gastroenteritis. Microbiology. 1994; 140(pt 12): 3441-9.
  11. Ceelen L, Decostere A, Verschraegen G, Ducatelle R, Haesebrouck F. Prevalence of Helicobacter pullorum among patients with gastrointestinal disease and clinically healthy persons. J Clin Microbiol. 2005; 43(6): 2984-6.
  12. Javed K, Gul F, Abbasi R, Zaidi R A, Noreen Z, Bokhari H, et al. Prevalence and role of Type six secretion system in pathogenesis of emerging zoonotic pathogen Helicobacter pullorum from retail poultry. Avian Pathol. 2019; 48(6): 557-63.
  13. Manfreda G, Parisi A, Lucchi A, Zanoni R G, De Cesare A. Prevalence of Helicobacter pullorum in conventional, organic, and free-range broilers and typing of isolates. Appl Environ Microbiol. 2011; 77(2): 479-84.
  14. Manfreda G, Rossi M, Sanguinetti V, Gavioli R, Lozito P. Prevalence of Helicobacter pullorum in broiler chickens reared in intensive and extensive farms. World Poult Sci J. 2006; 62: 551-2.
  15. González A, Piqueres P, Moreno Y, Cañigral I, Owen R J, Hernández J, et al. A novel real-time PCR assay for the detection of Helicobacter pullorum-like organisms in chicken products. Int Microbiol. 2008; 11(3): 203-8.
  16. Abdel Hameed K G, Slender G. Prevalence of H.pullorum in Egyptian hen, s eggs and in vitro suseptibility to different antimicrobial agents. Anim Sci Papers Reports. 2011; 29: 257-64.
  17. Wai S S, Abdul-Aziz S, Bitrus A A, Zunita Z, Abu J. Helicobacter pullorum in Broiler Chickens and the Farm Environment: A One Health Approach. Int J One Heal. 2019; 5(3): 20–5.
  18. Abd El-Ghany W A. Helicobacter pullorum: A potential hurdle emerging pathogen for public health. J Infect Dev Ctries. 2020; 14(11):1225–30.
  19. Steinbrueckner B, Haerter G, Pelz K, Weiner S, Rump J, Deissler W, et al. Isolation of Helicobacter pullorum from Patients with Enteritis. Scandina J infect Disease. 1997; 29(3): 315–8.
  20. Gholami-Ahangaran M, Azargoon R. PCR Amplification of Helicobacter pullorum 16S rRNA gene in cecal content of pet birds. Kafkas Uni Vet Fakult Dergisi. 2015; 21(2): 283-5.
  21. Zanoni R G, Piva S, Rossi M, Pasquali F, Lucchi A, De Cesare A, et al. Occurrence of Helicobacter pullorum in turkeys. Vet Microbiol. 2011; 149(3-4): 492-6.
  22. Javed S, Gul F, Javed K, Bokhari H. Helicobacter pullorum: An emerging zoonotic pathogen. Front Microbiol. 2017; 8: 604.
  23. Jamshidi A, Bassami R M, Salami H, Mohammadi S. Isolation and identification of Helicobacter pullorum from caecal content of broiler chickens in Mashhad, Iran. Iran J Vet Res. 2014; 15(2): 179-82.
  24. Behroo S, Javadi A, Rad MG. Helicobacter pullorum prevalence in patients with gastroenteritis in humans and chicken in the province of Ardabil in 2014. Indian J Fundam Appl Life Sci. 2015; 5(2): 87-94.
  25. Mohamed M, Ragab I, Shahata M, EI-Refaie M. Helicobacter pullorum among poultry in ASSIUT-Egypt: Genetic characterization, virolence and MIC. Int J Poult Sci. 2010; 9(6): 521-6.
  26. Kahraman B B, Seyyal A K. Investigation of Helicobacter pullorum occurrence in chicken in the marmara region of Turkey. Istanbul Univ Vet Fak Derg. 2013; 39(1): 63-6.
  27. Shah P M, Schäfer V, Knothe H. Medical and Veterinary Use of Antimicrobial Agents: Implications for Public Health. A Clinician’s View on Antimicrobial Resistance. Vet Microbiol. 1993; 35(3-4): 269–74.
  28. Ceelen M A, Haesebrouck F. In vitro susceptibility of Helicobacter pullorum strains to different antimicrobial agents. Microb Drug Resist. 2005; 11(2): 122-6.
  29. Bello N, Francino O, Sánchez A. Isolation of genomic DNA from feathers. J Vet Diagn Invest. 2001; 13(2): 162-4.
  30. Clinical and Laboratory Standards Institute (CLSI) performance standards for antimicrobial susceptibility testing. M100-S28, 2018, Wayne, P A.
  31. Hosseinzadeh S, Mardani K, Aliakbarlu J, Ghorbanzadehghan M. Occurrence of Campylobacter in chicken wings marketed in the northwest of Iran. I Food Research J. 2015; 22(1): 41-5.
  32. Fox J G, Chien C C, Dewhirst F E, Paster B J, Shen Z, Melito P L, et al. Helicobacter canadensis sp. nov. isolated from humans with diarrhea as an example of an emerging pathogen. J Clin Microbiol. 2000; 38(7): 2546-9.
  33. Matsuda M, Moore JE. Urease-positive thermophilic Campylobacter species. Appl Environ Microbiol. 2004; 70(8): 4415-8.