title: Molecular genetic analysis of Dichelobacter nodosus proteases AprV2/B2, AprV5/B5 and BprV/B in clinical material from European sheep flocks.
abstract: Dichelobacter nodosus, the etiological agent of ovine footrot, exists both as virulent and as benign strains, which differ in virulence mainly due to subtle differences in the three subtilisin-like proteases AprV2, AprV5 and BprV found in virulent, and AprB2, AprB5 and BprB in benign strains of D. nodosus. Our objective was a molecular genetic epidemiological analysis of the genes of these proteases by direct sequence analysis from clinical material of sheep from herds with and without history of footrot from 4 different European countries. The data reveal the two proteases known as virulent AprV2 and benign AprB2 to correlate fully to the clinical status of the individuals or the footrot history of the herd. In samples taken from affected herds, the aprV2 gene was found as a single allele whereas in samples from unaffected herds several alleles with minor modifications of the aprB2 gene were detected. The different alleles of aprB2 were related to the herds. The aprV5 and aprB5 genes were found in the form of several alleles scattered without distinction between affected and non-affected herds. However, all different alleles of aprV5 and aprB5 encode the same amino acid sequences, indicating the existence of a single protease isoenzyme 5 in both benign and virulent strains. The genes of the basic proteases BprV and BprB also exist as various alleles. However, differences found in samples from affected versus non-affected herds do not reflect the currently known epitopes that are attributed to differences in biochemical activity. The data of the study confirm the prominent role of AprV2 in the virulence of D. nodosus and shed a new light on the presence of the other protease genes and their allelic variants in clinical samples.
authors: Institute of Veterinary Bacteriology, Vetsuisse-Faculty, University of Bern, Postfach, Länggassstrasse 122, CH-3001 Bern, Switzerland; Clinic for Ruminants, Department of Clinical Veterinary Medicine, Vetsuisse-Faculty, University of Bern, Postfach, Bremgartenstrasse 109a, CH-3001 Bern, Switzerland.
authors: Anna
authors: A
authors: Stäuble
authors: Clinic for Ruminants, Department of Clinical Veterinary Medicine, Vetsuisse-Faculty, University of Bern, Postfach, Bremgartenstrasse 109a, CH-3001 Bern, Switzerland.
authors: Adrian
authors: A
authors: Steiner
authors: Institute of Veterinary Bacteriology, Vetsuisse-Faculty, University of Bern, Postfach, Länggassstrasse 122, CH-3001 Bern, Switzerland; University College of Northern Denmark, UCN, Aalborg, Denmark.
authors: Lea
authors: L
authors: Normand
authors: Institute of Veterinary Bacteriology, Vetsuisse-Faculty, University of Bern, Postfach, Länggassstrasse 122, CH-3001 Bern, Switzerland.
authors: Peter
authors: P
authors: Kuhnert
authors: Institute of Veterinary Bacteriology, Vetsuisse-Faculty, University of Bern, Postfach, Länggassstrasse 122, CH-3001 Bern, Switzerland. Electronic address: joachim.frey@vetsuisse.unibe.ch.
authors: Joachim
authors: J
authors: Frey
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D016000:Cluster Analysis; D020376:Dichelobacter nodosus / Q000145:classification / Q000201:enzymology* / Q000235:genetics* / Q000472:pathogenicity; D010450:Endopeptidases / Q000235:genetics*; D005060:Europe; D005535:Foot Rot / Q000382:microbiology*; D016905:Gram-Negative Bacterial Infections / Q000382:microbiology / Q000662:veterinary*; D008969:Molecular Sequence Data; D012697:Serine Endopeptidases / Q000235:genetics; D012756:Sheep; D012757:Sheep Diseases / Q000382:microbiology*; D014774:Virulence / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D010450:Endopeptidases; C097834:B2 protease, Dichelobacter nodosus; D012697:Serine Endopeptidases; C097833:V2 protease, Dichelobacter nodosus
keywords: Clinical scores; Footrot; Genetic alleles; Subtilisin-like proteases; Virulence