Human visceral (VL, also known as Kala-azar) and cutaneous (CL) leishmaniasis are important infectious diseases affecting countries in East Africa that remain endemic in several regions of Ethiopia. The transmission and epidemiology of the disease is complicated due to the complex life cycle of the parasites and the involvement of various Leishmania spp., sand fly vectors, and reservoir animals besides human hosts. Particularly in East Africa, the role of animals as reservoirs for human VL remains unclear. Isolation of Leishmania donovani parasites from naturally infected rodents has been reported in several endemic countries; however, the status of rodents as reservoirs in Ethiopia remains unclear. Here, we demonstrated natural Leishmania infections in rodents. Animals were trapped in 41 localities of endemic and non-endemic areas in eight geographical regions of Ethiopia and DNA was isolated from spleens of 586 rodents belonging to 21 genera and 38 species. Leishmania infection was evaluated by real-time PCR of kinetoplast (k)DNA and confirmed by sequencing of the PCR products. Subsequently, parasite species identification was confirmed by PCR and DNA sequencing of the 18S ribosomal RNA internal transcribed spacer one (ITS1) gene. Out of fifty (8.2%) rodent specimens positive for Leishmania kDNA-PCR and sequencing, 10 were subsequently identified by sequencing of the ITS1 showing that five belonged to the L. donovani complex and five to L. tropica. Forty nine kDNA-positive rodents were found in the endemic localities of southern and eastern Ethiopia while only one was identified from northwestern Ethiopia. Moreover, all the ten ITS1-positive rodents were captured in areas where human leishmaniasis cases have been reported and potential sand fly vectors occur. Our findings suggest the eco-epidemiological importance of rodents in these foci of leishmaniasis and indicate that rodents are likely to play a role in the transmission of leishmaniasis in Ethiopia, possibly as reservoir hosts.

Department of Microbiology and Molecular Genetics The Institute for Medical Research Israel Canada The Kuvin Centre for the Study of Infectious and Tropical Diseases The Hebrew University Hadassah Medical School The Hebrew University of Jerusalem Jerusalem 91120 Israel

Department of Microbiology Immunology and Parasitology Faculty of Medicine Addis Ababa University P O Box 9086 Addis Ababa Ethiopia

Department of Parasitology Faculty of Science Charles University Prague Vinicna 7 128 44 Prague 2 Czech Republic

Department of Zoological Science Addis Ababa University Addis Ababa Ethiopia

Department of Zoology Faculty of Science Charles University Prague Vinicna 7 128 44 Prague 2 Czech Republic

Institute of Vertebrate Biology Academy of Sciences of the Czech Republic 675 02 Studenec 122 Czech Republic

School of Veterinary Medicine Hebrew University P O Box 12 Rehovot 76100 Israel

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