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Journal of Clinical Microbiology, November 2003, p. 5167-5172, Vol. 41, No. 11
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.11.5167-5172.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detection of Gallibacterium spp. in Chickens by Fluorescent 16S rRNA In Situ Hybridization

Anders Miki Bojesen,1* Henrik Christensen,1 Ole Lerberg Nielsen,2 John Elmerdahl Olsen,1 and Magne Bisgaard1

Department of Veterinary Microbiology,1 Department of Pharmacology and Pathobiology, Laboratory of Veterinary Pathology, The Royal Veterinary and Agricultural University, DK-1870 Frederiksberg C, Copenhagen, Denmark2

Received 18 April 2003/ Returned for modification 27 July 2003/ Accepted 11 August 2003

Gallibacterium has recently been included as a new genus of the family Pasteurellaceae Pohl 1981, which encompasses bacteria previously reported as Pasteurella anatis, "Actinobacillus salpingitidis," and avian Pasteurella haemolytica-like organisms. So far, identification has exclusively relied on phenotypic characterization. We present a method based on a cyanine dye 3.18-labeled in situ hybridization probe targeting 16S rRNA to allow specific detection of bacteria belonging to the genus Gallibacterium. The probe, GAN850, showed no cross-reactivity to 25 other poultry-associated bacterial species, including members of the families Pasteurellaceae, Enterobacteriaceae, and Flavobacteriaceae, when cross-reactivities were evaluated by whole-cell hybridization. The probe was further evaluated by hybridization to formalin-fixed spleen and liver tissues from experimentally infected chickens, in which it proved to be useful for the detection of Gallibacterium. Additionally, determination of the spatial distribution and the host cell affiliation of Gallibacterium at various times during the infection process was possible. In conclusion, the in situ hybridization technique described may be of use as a diagnostic tool as well as for studies to elucidate the pathogenesis of Gallibacterium infections in chickens.

* Corresponding author. Mailing address: Department of Veterinary Microbiology, The Royal Veterinary and Agricultural University, 4 Stigbøjlen, DK-1870 Frederiksberg C, Copenhagen, Denmark. Phone: 45 3528 2781. Fax: 45 3528 2757. E-mail: miki{at}kvl.dk.

Journal of Clinical Microbiology, November 2003, p. 5167-5172, Vol. 41, No. 11
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.11.5167-5172.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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