Genetic Characterization of Campylobacter Jejuni and C. coli Isolated From Broilers Using flaA PCR-Restriction Fragment Length Polymorphism Method in Shiraz, Southern Iran

Abstract

Background:Thermophilic campylobacters, particularly Campylobacter jejuni and C. coli are the main agents of human campylobacteriosis. Campylobacter contaminated chicken products is the most important source of foodborne gastroenteritis. Evaluation of genetic diversity among Campylobacter population is critical for understanding the epidemiology of this bacterium and developing effective control strategies against Campylobacter infections and other related disorders.Objectives:The aim of this study was to investigate the polymorphism of thermophilic Campylobacter isolated from broiler fecal samples in Shiraz, southern Iran.Materials and Methods:Ninety Campylobacter isolates were recovered from broiler feces using enrichment process followed by cultivation method. The isolates were species typing on the basis of polymerase chain reaction (PCR) detection of 16SrRNA and multiplex PCR for determining two thermophilic species. To evaluate strain diversity of thermophilic Campylobacter isolates, flaA PCR-Restriction Fragment Length Polymorphism (RFLP) was performed using DdeI restriction enzyme.Results:All 90 Campylobacter isolates confirmed by m-PCR were successfully typed using flaA-PCR-RFLP. Eleven different types were defined according to flaA-typing method and the RFLP patterns were located at three separate clusters in RFLP image analysis dendrogram.Conclusions:Campylobacter jejuni isolates significantly showed more variety than C. coli isolates. A relatively low genetic diversity existed among C. jejuni and C. coli isolated from broilers in Shiraz, southern Iran. In our knowledge, this was the first report of genetic diversity among broiler originated human pathogen thermophilic campylobacters in Shiraz, southern Iran.

Typing of Campylobacter jejuni isolated from poultry on the basis of flaA-RFLP by various restriction enzymes

“…Dde I (D.I.−0.9258) was found most efficient in discriminating C. jejuni isolates as has been reported by many workers ( Hiett et al., 2006 , Khoshbakht et al., 2015 , Rajagunalan et al., 2014 , Vinueza-Burgos et al., 2017 ). The other enzymes i.e Hinf I (D.I.−0.6977) and Dpn II (D.I.−0.8427) were less discriminatory as compared to Dde I.…”

“…Flagellin gene locus of C. jejuni contains two flagellin genes ( fla A and fla B), which are highly conserved with variable regions interspersed between them ( Khoshbakht et al., 2015 , Thomrongsuwannakij et al., 20 ). This locus is suitable for restriction fragment length polymorphism (RFLP) analysis of PCR products.…”

RFLP analysis of flagellin (Fla) gene of Campylobacter jejuni from ovines of Kashmir, India

“…In the present study, an identical pattern was observed on digestion of amplified fla A gene of C. jejuni and C. coli isolates indicating no diversity among isolates. The pattern observed in the study belong to cluster III (Pattern I) based on dendogram study of restriction profiles of fla A gene of virulent strains of C. jejuni and C. coli reported by workers (Khoshbakht, Tabatabaei, Hosseinzadeh, Shirzad, & Seifi, ). Of the various restriction enzymes, Alu I, Dde I, Hin fI, Eco RI, and Pst I enzymes used for RE studies, Hin fI is not able to discriminate clearly among the isolates, whereas Alu I produces bands that are too small to be analyzed.…”

Isolation, Identification and Molecular Detection of Zoonotic Campylobacter jejuni Isolated from Mutton and Beef Samples

“…(5-CTATTTTATTTTTGAGTGCTTGTG-3) and reverse primers (5-GCTTTATTTGCC ATTTGTTTTATTA-3) (Khoshbakht et al, 2015).…”