Detection and quantification of Leishmania braziliensis in ectoparasites from dogs.

American cutaneous leishmaniasis (ACL) is a disease caused by different species of Leishmania protozoa, Leishmania braziliensis being the main species found in Brazil. In this study, two rural areas in Pernambuco, northeastern Brazil, where ACL is endemic, were selected. Genomic DNA was extracted from canine ectoparasites (ticks, fleas, and lice) and tested using a conventional PCR and a quantitative real time PCR. A total of 117 ectoparasites were collected, being 50 (42.74%) of them positive for L. braziliensis (in at least one PCR protocol), with a mean parasite load of 14.14 fg/µL. Furthermore, 46 (92.00%) positive ectoparasites were collected from positive dogs and 4 (8.00%) from negative ones. This study reports the detection of L. braziliensis DNA in ectoparasites, but does not prove their vector competence. Certainly, experimental transmission studies are necessary to assess their role, if any, in the transmission of Leishmania parasites to dogs.

Detection of Leishmania infantum in animals and their ectoparasites by conventional PCR and real time PCR.

Visceral leishmaniosis (VL) is a parasitic disease caused by Leishmania infantum, which is primarily transmitted by phlebotomine sandflies. However, there has been much speculation on the role of other arthropods in the transmission of VL. Thus, the aim of this study was to assess the presence of L. infantum in cats, dogs and their ectoparasites in a VL-endemic area in northeastern Brazil. DNA was extracted from blood samples and ectoparasites, tested by conventional PCR (cPCR) and quantitative real time PCR (qPCR) targeting the L. infantum kinetoplast DNA. A total of 280 blood samples (from five cats and 275 dogs) and 117 ectoparasites from dogs were collected. Animals were apparently healthy and not previously tested by serological or molecular diagnostic methods. Overall, 213 (76.1 %) animals and 51 (43.6 %) ectoparasites were positive to L. infantum, with mean parasite loads of 795.2, 31.9 and 9.1 fg in dogs, cats and ectoparasites, respectively. Concerning the positivity between dogs and their ectoparasites, 32 (15.3 %) positive dogs were parasitized by positive ectoparasites. The overall concordance between the PCR protocols used was 59.2 %, with qPCR being more efficient than cPCR; 34.1 % of all positive samples were exclusively positive by qPCR. The high number of positive animals and ectoparasites also indicates that they could serve as sentinels or indicators of the circulation of L. infantum in risk areas.