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Applied and Environmental Microbiology, May 2002, p. 2172-2178, Vol. 68, No. 5
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.5.2172-2178.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Assessment of the Genetic Diversity among Arcobacters Isolated from Poultry Products by Using Two PCR-Based Typing Methods

Kurt Houf,1* Lieven De Zutter,1 Jan Van Hoof,1 and Peter Vandamme2

Department of Veterinary Food Inspection, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke,1 Department of Microbiology, Faculty of Sciences, Ghent University, 9000 Ghent, Belgium2

Received 26 October 2001/ Accepted 6 February 2002

In this study, enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and randomly amplified polymorphic DNA PCR (RAPD-PCR) were optimized for characterization of Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. In addition, a simple and rapid DNA extraction method was tested for use in both typing procedures. Both methods had satisfactory typeability and discriminatory power, but the fingerprints generated with ERIC-PCR were more reproducible and complex than those obtained with RAPD-PCR. The use of nondiluted boiled cell suspensions as DNA templates was found to be very useful in ERIC-PCR. Characterization of large numbers of Arcobacter isolates is therefore preferably performed by the ERIC-PCR procedure. Isolates for which almost identical ERIC fingerprints are generated may subsequently be characterized by RAPD-PCR, although adjustment and standardization of the amount of the DNA template are necessary. In the second part of this study, the genotypic diversity of arcobacters present on broiler carcasses was assessed by using both typing methods. A total of 228 cultures from 24 samples were examined after direct isolation and enrichment. The isolates were identified by using a multiplex PCR as A. butzleri (n = 182) and A. cryaerophilus (n = 46). A total of 131 types (91 A. butzleri types and 40 A. cryaerophilus types) were discerned without discordance between the two typing techniques. The analysis of the poultry isolates showed that poultry products may harbor not only more than one species but also multiple genotypes. All genotypes were confined to one poultry sample, and only three genotypes were found after simultaneous enrichment and direct isolation. These results demonstrate that different outcomes can be obtained in epidemiological studies depending on the isolation procedure used and the number of isolates characterized.

* Corresponding author. Mailing address: Department of Veterinary Food Inspection, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium. Phone: 32 09 264 74 51. Fax: 32 09 264 74 91. E-mail: kurt.houf{at}rug.ac.be.

Applied and Environmental Microbiology, May 2002, p. 2172-2178, Vol. 68, No. 5
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.5.2172-2178.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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