African Journal of
Microbiology Research

  • Abbreviation: Afr. J. Microbiol. Res.
  • Language: English
  • ISSN: 1996-0808
  • DOI: 10.5897/AJMR
  • Start Year: 2007
  • Published Articles: 5112

Full Length Research Paper

Use of a combined cultural-molecular method for isolation and identification of Campylobacter from broiler chicken in Morocco

Charrat Nadia*
  • Charrat Nadia*
  • Laboratory of Microbiology and Molecular Biology, Department of Food Microbiology, Research Laboratory and Medical Analysis (LRAM), Fraternal of the Royal Gendarmerie, Rabat, Morocco.
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El Fahime Elmostafa
  • El Fahime Elmostafa
  • Laboratory of Sequencing and Genotyping, National Center for the Scientific and Technical Research (CNRST), Rabat, Morocco.
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Filali-Maltouf Abdelkarim
  • Filali-Maltouf Abdelkarim
  • Laboratory of Microbiology and Molecular Biology, Faculty of Science (FSR), University Mohammed V, Rabat, Morocco.
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  •  Received: 23 November 2016
  •  Accepted: 31 January 2017
  •  Published: 21 February 2017

Abstract

Campylobacters are very important foodborne pathogens that raise the interest of food processors, researchers, as well as consumers and all stakeholders. Their contaminations can result in life threatening disorders, potentially leading to chronic sequelae such as Reiter’s and Guillain-Barré syndromes or Crohn’s Disease. Poultry has been identified as the main and most common reservoirs for Campylobacter. A survey was conducted in Morocco, from 2009 to 2012 to estimate the prevalence of Campylobacter in broiler chicken from about fifteen Moroccan cities that were most involved in the breeding of broilers. In this study, thermotolerant Campylobacter spp. were detected and identified from 165 samples by both cultural methods and molecular approaches based on polymerase chain reaction  and 16S ribosomal RNA (rRNA) gene sequencing. The species were also genotyped by Restriction Fragment Length Polymorphism analysis. The conventional culture methods identified 97% of samples as positive for Campylobacter spp. The molecular approach based on 16S rRNA gene could not distinguish between C. jejuni, C. coli and C. lari. However, gyrB gene RFLP, allowed a good discrimination between the three species of Campylobacter. These results were also confirmed by Matrix Assisted Laser Desorption Ionization-Time of Flight and Mass Spectrometry proteomic profiling determination using libraries in the BioTyper 2.3 software. The present study identifies C. jejuni as the major source of contamination of poultry carcasses in Morocco.

Key words: Campylobacter, chicken, 16S rRNA gene, gyrB gene, sequencing, RFLP-PCR, MALDI-TOF MS.