Dogs' parasite and zoonotic risk: from old to new "emergencies" in the North-West of Italy.

Toxocariasis due to soil contamination from dog and cat faeces has been long described and represents one of the zoonotic risk linked with pets presence in human settlements. Soil samples were collected from private backyards and school playgrounds in Turin and tested for the presence of Toxocara spp. eggs. Samples from dogs and cats living in the same area were also analysed and our results seem to indicate a decrease in soil contamination respect to a survey carried out in 1985. Considering that recently new foci of Canine Leishmaniosis and the presence of competent sand fly vectors have also been reported in the North-West of Italy, a survey was carried out on dogs and humans living in Asti province. To assess the risk of local Leishmania infantum transmission between dog and humans, samples were also analysed by Restriction Fragment Length Polymorphism (PCR-RFLP). Our results have shown that more than 10% of autochthonous dogs and human being living in this previously non-endemic area have been infected by L. infantum. The identity of PCR-RFLP patterns from 3 human clinical cases and from the dogs of one of them allows us to confirm the autochthonous origin of these cases.

Characterization of Leishmania infantum strains in blood samples from infected dogs and humans by PCR-RFLP.

Characterization of Leishmania infantum is based on zymodeme analysis, which requires parasite isolation and therefore is not routinely employed. Moreover, the majority of strains in the Mediterranean Basin belong to zymodeme MON-1, and this is a major limitation for this technique in epidemiological studies in this region. We developed a PCR-RFLP method based on kDNA amplification, which was able to discriminate L. infantum strains directly from peripheral blood. Twenty-eight samples were tested with this technique: four obtained from promastigote cultures, and 24 collected from dogs (18) and human donors (six) from traditionally endemic and newly endemic areas of northwestern Italy. Extracted DNAs were amplified using RV1-RV2 primers and PCR products were digested using two restriction enzymes separately: BsiY I and Mlun NI. Some patterns were specific to certain areas. In particular, the identity of PCR-RFLP patterns from a human patient from a newly endemic area and three dogs allow the confirmation of the autochthonous origin of this case. This approach could be applied to epidemiological studies in order to trace the diffusion of L. infantum within dog populations, as well as its transmission to humans.