Introduction: Leishmania major is the causative agent of zoonotic cutaneous leishmaniasis (ZCL), and great gerbils are the main reservoir hosts in Iran. Abarkouh in central Iran is an emerging focal point for which the reservoir hosts of ZCL are unclear. This research project was designed to detect any Leishmania parasites in different wild rodent species. Methods: All rodents captured in 2011 and 2012 from Abarkouh district were identifi ed based on morphological characteristics and by amplifi cation of the rodent cytochrome b (Cyt b) gene. To detect Leishmania infection in rodents, deoxyribonucleic acid (DNA) of each ear was extracted. Internal transcribed spacer-ribosomal deoxyribonucleic acid (ITS-rDNA), microsatellites, kinetoplast deoxyribonucleic acid (kDNA) and cytochrome b genes of Leishmania parasites were amplifi ed by polymerase chain reaction (PCR). Restriction fragment length polymorphism (RFLP) and sequencing were employed to confi rm the Leishmania identifi cation. Results: Of 68 captured rodents in the region, 55 Rhombomys opimus were identifi ed and nine Leishmania infections (9/55) were found. In addition, eight Meriones libycus and two Tatera indica were sampled, and one of each was confi rmed to be infected. Two Meriones persicus and one Mus musculus were sampled with no infection. Conclusions: The results showed that all 11 unambiguously positive Leishmania infections were Leishmania major. Only one haplotype of L. major (GenBank access No. EF413075) was found and at least three rodents R. opimus, M. libycus and T. indica-appear to be the main and potential reservoir hosts in this ZCL focus. The reservoir hosts are variable and versatile in small ZCL focal locations.

Background: Leishmania infantum parasites are the main causative agents of visceral leishmaniasis that threaten a wide range of humans and canines in Iran. Objectives: Our aim was to survey Leishmania parasite species and simultaneous comparison of canine organs in endemic areas for the diagnosis of visceral leishmaniasis using the ITS-rDNA gene, sequencing, and phylogenetic analysis. Methods: In this study, sampling was done with vacuum tubes containing EDTA from blood and sterile swabs from the snout and conjunctiva of asymptomatic sheepdogs (n = 37) using a non-invasive method in north Khorasan, northeastern Iran, from 28 July to 4 August 2018. The DNA of collected samples was extracted, amplified, and sequenced by targeting the ITS-rDNA gene. To demonstrate the taxonomic status of Leishmania spp., sequences were subjected to phylogenetic analysis based on the maximum likelihood method. Results: We obtained 37 samples from asymptomatic dogs of which, 10 dogs were definitely diagnosed with L. infantum and one dog infected with L. tropica. The blood (n = 8) and right conjunctiva (n = 6) samples were the most infected samples. The highest number of infections in dogs was in the age group of 5-10 years indicating that this group is more sensitive to visceral leishmaniasis in this region. Conclusions: The current findings indicate that non-invasive sampling and molecular methods are reliable and suitable in the detection of visceral leishmaniasis. This is the first report of the visceral involvement of a shepherd dog with L. tropica in northeastern Iran. The remarkable occurrence of visceral leishmaniasis (29.7%) in asymptomatic sheepdogs reflects a health alert to conduct the surveillance and monitoring of susceptible individuals/reservoirs in the region.

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