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Objective After the epidemics of L. donovani complex in 2004/05 in human patients, to investigate the presence of antibodies against L. donovani in domestic animals in north-west Ethiopia. Methods Two hundred and three domestic animals were screened. Serum and biopsy samples were collected. A modified direct agglutination test ( DAT) for canine reservoirs was used to screen serum samples at ≥ 1:320 cut-off titre. Giemsa stain and culture on Novy macNeal Nicolae ( NNN) media were used for biopsy samples. Pre-tested questionnaires were used to elicit information on potential risk factors. Results Antibody against L. donovani in domestic animals was detected in 30.5% of animals. The highest seropositivity rates were 41.9% in cattle, 40% in dogs, 33.3% in donkeys, 10% in goats and 4.8% in sheep. No Leishmania parasite was isolated from spleen, liver, skin snip and exudates, bone marrow or lymph node of dogs. Dogs owned by households with history of kala-azar treatment and humans sharing the house with cattle were more affected by visceral leishmaniasis ( P < 0.05). Conclusion This study showed a high serological prevalence of leishmaniasis in domestic animals. Their role in the epidemiology of visceral leishmaniasis remains unclear. [ABSTRACT FROM AUTHOR]