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A cross-sectional serosurvey using Leishmania infantum ELISA was performed on 445 cats living in ecoregions around the Northwestern Mediterranean basin; 58 cats from an area of the US where leishmaniasis is not endemic were used as negative controls. ELISA results were further confirmed in 69 cats by Western blot (WB). Finally, 76 of them were also tested for FeLV and FIV. Seroprevalence by ELISA-prot A was 6.29%, and that by ELISA-IgG was 5.25%. Positive cat sera recognized patterns of polypeptides in WB, including L. infantum-specific antigenic fractions. There was no association with retroviruses. Leishmania-specific antibodies are prevalent in cats living in ecoregions around the Northwestern Mediterranean basin; thus, leishmaniasis must be included in the differential diagnosis of diseases in cats living in these ecoregions. Their role as peridomestic reservoirs for L. infantum needs further characterization, but it could be hypothesized that the cat is a secondary reservoir host, rather than an accidental one.

The extent of cryptic leishmaniasis in blood donors from a Spanish endemic area, (Eivissa Island) was studied using various immunological and parasitological methods. Sera from 656 blood donors were analysed: 16 (2.4%) were positive by ELISA and 50 (7.6%) by Western blot. Peripheral blood mononuclear cells (PBMC) and buffy coat (BC) samples, were analyzed by culture and nested-PCR. DNA of L. infantum was amplified in 27 (22.1%) of 122 PBMC. Parasites were isolated in 3 (4.5%) of 67 BC cultures and the strains were identified as L. infantum zymodeme MON-28. No parasites were isolated in PBMC culture. After 12 months, a second blood sample was obtained from 18 blood donors who were positive by nested-PCR in the first extraction; nine of them remained positive. Delayed type hypersensitivity (DTH) tests on 15/67 donors (22.3%) were positive. Comparison of results obtained by ELISA, WB and DTH; ELISA, WB and nested-PCR and nested-PCR and BC culture showed a significant association (Pearson test, P < 0.05). L. infantum zyodeme MON-28 was identified in three strains isolated from asymptomatic donors, which suggests a low virulence capacity of these strains. The detection of Leishmania DNA in a high number of asymptomatic subjects supports the need to monitor it in blood donors endemic areas.

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