Poster communication in 4th Med-Vet-Net Annual Scientific Meeting 2015 (Global Challenges in Zoonoses: Combating Emerging Threats using a One-Health Approach)

October 8^th, 2015

Ortega-García MV., Moreno I., Dominguez M. and Garcia N.

In the Mediterranean basin, where Leishmaniasis is an endemic disease, Leishmania infantum is the main species implicated and the dog its main domestic reservoir. However, in recent years Leporidae have been described as competent reservoirs. A rapid and accurate identification method of L. infantum is essential to improve surveillance programs in wild animals and real-time PCR offers these advantages over conventional PCR.
The aim of the study is to validate a real-time PCR assay described for dogs in blood, to detect L. infantum in different samples of 226 wild rabbits and 70 hares (Leporidae) from Madrid area and compare it to nested PCR.
A total of 71 rabbits (32.13%, 95% CI: 25.97-38.28) and 8 hares (11.43%, 95% CI: 3.98-18.88), and 178 rabbits (87.68%, 95% CI: 83.16-92.21) and 47 hares (67.14%, 95% CI: 56.14-78.15) were positive in spleen and skin samples, respectively by real-time PCR; whereas, 5 rabbits (2%, 95% CI: 0-4) and no hares, and 26 rabbits (11.50%, 95% CI: 7.41-15.8) and 1 hare (1.45%, 95% CI: 0-4.27) were positive in spleen and skin samples, respectively by nested PCR. Furthermore, 21 rabbits (70%, 95% CI: 53.60-86.4) and 17 hares (56.67%, 95% CI: 38.93-74.4) were positive in hair samples by real-time PCR.
The assay is more sensitive than nested PCR, as it has been probed in dogs, with a limit of detection of 31, 35 pg/reaction of L. infantum genomic DNA (0.0854 parasites), but lower than it had in dogs (0.001 parasites), so it could be considered adequate for diagnosis of L.infantum infection in wild Leporidae

	Instituto Tecnológico La Marañosa (ITM). Ministerio de Defensa.
	Instituto Nacional de Técnica Aeroespacial (INTA). Ministerio de Defensa.
	Servicio de Inmunología. Centro Nacional de Microbiología (CNM). Instituto de Salud Carlos III (ISCIII).
	Servicio de Zoonosis Emergentes, de Baja Prevalencia y Agresivos Biológicos (NED). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM).