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Title: RESTRICTION ENZYME ANALYSIS AND RIBOTYPING DISTINGUISH BORDETELLA AVIUM AND BORDETELLA HINZII ISOLATES

Author
item Sacco, Randy
item Register, Karen
item Nordholm, Gwen

Submitted to: Epidemiology and Infection
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/9/1999
Publication Date: N/A
Citation: N/A

Interpretive Summary: Bordetella avium causes a respiratory infection in poultry known as coryza. A closely related organism, Bordetella hinzii, is also isolated from poultry. Diffentiating between these organisms has been primarily based on methods which examine proteins or fatty acids or more complicated genetic approaches. In this study, we have developed methods to easily distinguish hbetween these two organisms. Our results are based on the use of genetic material from these organisms. The methods that we have used are relatively easy to carry out compared to previous methods. Further, our methods may also be used as an identification system during outbreaks of coryza.

Technical Abstract: Fifty four bacterial strains previously identified as Bordetella avium or B. hinzii were characterized by restriction enzyme analysis (REA) and/or ribotyping. Twenty restriction endonucleases were evaluated for use in REA. Digestion of chromosomal DNA with Hinf I, followed by submarine electrophoresis in agarose gels and staining with ethidium bromide, produced DNA fragments in the 3.0- to 10-kb range which readily distinguished B. avium and B. hinzii isolates. Moreover, Hinf I digestion of chromosomal DNA from B. avium and B. hinzii strains produced eight and six distinct fingerprint profiles, respectively. Digestion with Dde I resulted in DNA fragments in the 3 to 23.1 kb molecular size range. While the fragments produced by Dde I digestion of B. avium and B. hinzii were not as readily distinguished as those produced by Hinf I, Dde I digestion further discriminated these Bordetella species producing twelve fingerprint tprofiles for B. avium and four profiles for B. hinzii. In addition, B. avium and B. hinzii isolates were readily distinguished by ribotyping with the restriction endonuclease Pvu II. The ribotype patterns produced by these two species of Bordetella were unique when compared to previously reported ribotype patterns for B. bronchiseptica isolates. The combination of DNA fingerprinting and ribotyping should provide valuable information in understanding the molecular epidemiology of Bordetella infections.