DETECTION OF PESTIVIRUS IN FECAL SAMPLES FROM WILD RUMINANTS BY PCR

Ruminant pestiviruses affect wild and domestic ruminants worldwide, causing reproductive disorders and severe economic losses. Usually serological, virological or PCR tests are carried out on serum or blood collected from captured live animals or from organs of dead animals. The aim of this work was to evaluate if fecal samples are suitable for detecting ruminant pestiviruses in wild ruminants. Fecal samples from red deer (Cervus elaphus, n=16) and Appenninic chamois (Rupicapra pyrenaica ornata, n=12) were collected from the environment. Samples were analyzed in a few days and aliquots were stored at 4ºC for 3 months and re-tested. Pooled fecal samples were diluted with PBS, were centrifuged, and the pellet was used for RNA extraction by silica membrane tech-nology. Real Time PCR was carried out to detect Pestivirus spp. Positive samples were used for PCR and sequencing of a 5’UTR sequence. All four pool samples from red deer and 3 out of 4 pool of chamois resulted positive and sequences showed the highest homology with BVDV-1. Positive samples were found also in samples refrigerated for 3 months. To our knowledge, this is the first report of amplification and sequencing of pestiviruses from feces of wild ruminants.