Comparative restriction enzyme mapping of Campylobacter jejuni isolates from turkeys and broilers based on flaA flagellar gene using HpyF3I endonuclease
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Turkeys and broilers have been identified as important reservoirs for Campylobacter jejuni which is of public health significance. The evaluation of the genotypes among C. jejuni strains within different reservoirs is critical for our understanding of the epidemiology of this infectious agent. The present study aimed to compare the genetic diversity and differences of C. jejuni isolates from turkeys and broilers using flagellin PCR-RFLP typing (flaA typing) technique, in terms of the ease of use and discriminatory power. Sixty C. jejuni isolates were detected biochemically and confirmed by duplex-PCR from turkeys and broilers (30 strains from each bird species). Then, a flaA gene fragment (1725 bp) of C. jejuni isolates was amplified and amplicons were digested with HpyF3I enzyme. Restriction analysis by HpyF3I gave four different flaA patterns (H1, H2, H3, H4) among all tested C. jejuni isolates. In broiler isolates, all four patterns were observed but in turkey isolates, only H2 and H4 patterns were present. The results clearly demonstrated that distribution of the flaA typing patterns differed depending on the host species (broiler/turkey). H1 and H3 flaA types are more prevalent in broiler than turkey isolates, while H2 type is significantly more prevalent within isolates from turkey (p < 0.05). The flaA typing technique by digestion with HpyF3I enzyme can almost give us a clue to the source of infection in local outbreaks.
The authors gratefully acknowledge the staff of laboratory of microbiology, faculty of veterinary medicine, University of Semnan (Iran), especially Mrs. Behnaz Raeisian and Mrs. Birgani Farhani. This study was carried out in partial fulfillment of the requirements for a DVM student’s thesis (Elham Atefi Tabar). We deeply thank Dr. Masoumeh Estaji (Allameh Tabataba’i University) for English revision of the manuscript.
We acknowledge Semnan University for funding the study.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- El-Adawy H, Hotzel H, Tomaso H, Neubauer H, Taboada EN, Ehricht R, Hafez HM (2013) Detection of genetic diversity in Campylobacter jejuni isolated from a commercial turkey flock using flaA typing, MLST analysis and microarray assay. PLoS One 8:e51582. https://doi.org/10.1371/journal.pone.0051582 CrossRefGoogle Scholar
- Ertaş HB, Çetinkaya B, Muz A, Öngör H (2004) Genotyping of broiler-originated Campylobacter jejuni and Campylobacter coli isolates using fla typing and random amplified polymorphic DNA methods. Int J Food Microbiol 94:203–209. https://doi.org/10.1016/S0168-1605(03)00312-X
- Fitzgerald C, Helsel LO, Nicholson MA, Olsen SJ, Swerdlow DL, Flahart R, Sexton J, Fields PI (2001) Evaluation of methods for subtyping Campylobacter jejuni during an outbreak involving a food handler. J Clin Microbiol 39:2386–2390. https://doi.org/10.1128/JCM.39.7.2386-2390.2001 CrossRefGoogle Scholar
- Hajieh GS, Mohammad J, Ahad H, Tahmineh N, Ali S, Ebrahim R (2011) The prevalence of bacterial contamination of table eggs from retails markets by Salmonella spp., Listeria monocytogenes, Campylobacter jejuni and Escherichia coli in Shahrekord, Iran. Jundishapur J Microbiol 4(4):249–253Google Scholar
- Lauwers S, Penner J (1984) Serotyping Campylobacter jejuni and Campylobacter coli on the basis of thermostable antigensGoogle Scholar
- Nachamkin I, Bohachick K, Patton C (1993) Flagellin gene typing of Campylobacter jejuni by restriction fragment length polymorphism analysis. J Clin Microbiol 31:1531–1536Google Scholar
- Organization WH (2015) Investing to overcome the global impact of neglected tropical diseases: third WHO report on neglected tropical diseases: World Health OrganizationGoogle Scholar
- Osmundson TW, Eyre CA, Hayden KM, Dhillon J, Garbelotto MM (2013) Back to basics: an evaluation of NaOH and alternative rapid DNA extraction protocols for DNA barcoding, genotyping, and disease diagnostics from fungal and oomycete samples. Mol Ecol Resour 13:66–74. https://doi.org/10.1111/1755-0998.12031 CrossRefGoogle Scholar
- Ozawa M, Hiki M, Kawanishi M, Abo H, Kojima A, Asai T, Hamamoto S (2016) Molecular typing of fluoroquinolone-resistant Campylobacter jejuni isolated from broilers in Japan using multilocus sequence typing and pulsed-field gel electrophoresis. Foodborne Pathog Dis 13:1–7. https://doi.org/10.1089/fpd.2015.1975 CrossRefGoogle Scholar
- Patton CM, Wachsmuth I, Nachamkin I, Blaser M, Tompkins L (1992) Typing schemes: are current methods useful? American Society for Microbiology, WASHINGTON, DCGoogle Scholar
- Ragimbeau C, Schneider F, Losch S, Even J, Mossong J (2008) Multilocus sequence typing, pulsed-field gel electrophoresis, and fla short variable region typing of clonal complexes of Campylobacter jejuni strains of human, bovine, and poultry origins in Luxembourg. Appl Environ Microbiol 74:7715–7722. https://doi.org/10.1128/AEM.00865-08 CrossRefGoogle Scholar
- Stabler RA, Larsson JT, Al-Jaberi S, Nielsen EM, Kay E, Tam CC, Higgins CD, Rodrigues LC, Richardson JF, O’brien SJ (2013) Characterization of water and wildlife strains as a subgroup of Campylobacter jejuni using DNA microarrays. Environ Microbiol 15:2371–2383. https://doi.org/10.1111/1462-2920.12111 CrossRefGoogle Scholar
- Takahashi R, Shahada F, Chuma T, Okamoto K (2006) Analysis of Campylobacter spp. contamination in broilers from the farm to the final meat cuts by using restriction fragment length polymorphism of the polymerase chain reaction products. Int J Food Microbiol 110(3):240–245Google Scholar
- Vinueza-Burgos C, Wautier M, Martiny D, Cisneros M, Van Damme I, De Zutter L (2017) Prevalence, antimicrobial resistance and genetic diversity of Campylobacter coli and Campylobacter jejuni in Ecuadorian broilers at slaughter age. Poult Sci 96:2366–2374. https://doi.org/10.3382/ps/pew487 CrossRefGoogle Scholar
- Zbrun MV, Romero-Scharpen A, Olivero C, Rossler E, Soto LP, Rosmini MR, Sequeira GJ, Signorini ML, Frizzo LS (2013) Occurrence of thermotolerant Campylobacter spp. at different stages of the poultry meat supply chain in Argentina. New Zealand Vet J 61:337–343. https://doi.org/10.1080/00480169.2013.817294 CrossRefGoogle Scholar