Occurrence of Baylisascaris transfuga in wild populations of European brown bears (Ursus arctos) as identified by a new PCR method.

The European brown bear (Ursus arctos) is a species present in limited areas of Europe and several small populations are considered endangered. This species can be affected by a range of parasites. In particular, the genus Baylisascaris is an emerging parasite of wild animals which can cause severe larva migrans syndrome in aberrant hosts, which include 100 species of birds, mammals and also humans. Baylisascaris transfuga is the species reported from bears, and with the exception of a few laboratory trials, little is known about the capacity of this species to infect other animals. Furthermore, the identification of this species has traditionally been based on light microscopy, using either morphology of the adults at necropsy or detection of the eggs in faeces, which are methods limited by the experience and the efforts of the observer. The current work was aimed at developing a specific PCR to detect the parasite directly from faecal samples of naturally infected brown bears in the field, without the need for previous flotation. Using eggs and adults of B. transfuga collected in Croatia, we first developed a PCR to detect a portion of the second internal transcribed spacer region (ITS-2) of ribosomal DNA and then applied it to bear faecal samples spiked with a known number of B. transfuga eggs. We show here for the first time that this method allows the detection of a minimum of two Baylisascaris eggs in 25mg of faecal material, thus it demonstrates a high diagnostic capacity that could be applied to evaluate the prevalence of the parasite in faecal samples from wild populations of brown bears. Crown