[The use of in-situ hybridization for the detection of Brachyspira spp. in pigs].
Diagnosis of Brachyspira infections in swine and the differentiation of the involved bacteria is time-consuming and in most cases unsatisfactory. Detecting Brachyspira directly in the damaged Brachyspira of the large intestine could provide a direct correlation between histological lesionsa and bacterial growth. In this study we investigated whether in-situ hybridization (ISH) with a digoxigenin-labeled RNA-probe is a suitable method for detecting Brachyspira in the mucosa of the large intestine. Formalin-fixed and paraffin-embedded tissue sections of the large intestine from 78 pigs, which showed macroscopic and histological findings of Brachyspira-associated colitis, were stained with hematoxylin and eosin and Warthin-Starry silver impregnation and subjected to ISH. We used a RNA-probe with a length of 334bp, complementary to a part of the 23S rRNA of all members of the genus Brachyspira. All sections were treated with this anti-sense probe and with a sense control probe. 64 samples (82%) showed clearly positive ISH signals. Thus ISH is a suitable method for detecting Brachyspira directly within the lesions of the large intestine. The quantity of Brachyspira identified by ISH was always lower than by Warthin-Starry staining. Whether this reflects lower sensitivity of the ISH technique, or the fact that other bacteria with morphological similarities to Brachyspira were also stained by Warthin-Starry is unknown as yet. The present investigations provide a basis of further research developing specific probes to distinguish between pathogenic and non pathogenic Brachyspira species and probes detecting other bacteria with morphological similarity to Brachyspira.
Herzog
A M
AM
Institut für Pathologie und Gerichtliche Veterinärmedizin, Department für Pathobiologie, Veterinärmedizinische Universität Wien.
Maderner
A
A
Fragner
K
K
Weissenbock
H
H
D000818:Animals; D017400:Brachyspira / Q000235:genetics / Q000302:isolation & purification*; D017403:In Situ Hybridization / Q000379:methods / Q000662:veterinary*; D007413:Intestinal Mucosa / Q000382:microbiology; D015347:RNA Probes; D012329:RNA, Bacterial / Q000032:analysis / Q000235:genetics; D013045:Species Specificity; D013145:Spirochaetales Infections / Q000175:diagnosis / Q000662:veterinary*; D013194:Staining and Labeling; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000382:microbiology
D015347:RNA Probes; D012329:RNA, Bacterial