BackgroundThis study aimed to evaluate the occurrence of Leishmania spp. in dogs and cats from Botucatu, São Paulo state, and Campo Grande, Mato Grosso do Sul state, Brazil, by the association of three diagnostic tests: blood culture in liver infusion tryptose medium, immunofluorescent antibody test and polymerase chain reaction. Fifty blood samples of dogs and cats from the Center for Zoonosis Control in Campo Grande, an area endemic for canine visceral leishmaniasis, were collected randomly, as well as canine and feline blood samples from the Municipal Kennel and Animal Protection Association in Botucatu, currently considered a transmission-free, non-endemic area.ResultsOf the 50 dog blood cultures from Botucatu, three (6%) were positive and of the 50 cats, two (4%) were positive. In Campo Grande, 29 dog blood cultures (58%) were positive and all (100%) cats negative by this test. Polymerase chain reaction detected Leishmania spp. in 100% of dog and cat samples from Botucatu but found all the cats from Campo Grande to be negative. On the other hand, 36 dogs from Campo Grande were positive (72%) by the same technique. Immunofluorescent antibody test in Botucatu found 100% of dogs and cats non-reactive, while in Campo Grande, it detected positivity in 32 dogs (64%) and 15 cats (30%).ConclusionsThe results show the importance of not only continuous epidemiological surveillance in areas not endemic for leishmaniasis, but also research for accurate diagnosis of this zoonosis.

Toxoplasma gondii and Leishmania spp. are protozoan parasites that cause zoonosis, are distributed worldwide, and affect domestic and wild animals and humans. Cats are important in the epidemiology of toxoplasmosis because they are the only hosts that can excrete the environmentally resistant stage: the oocysts 1 .In 1999, visceral leishmaniasis in cats was fi rst recorded by Hervas et al. 2 in Spain. Since then, current cases represent a widespread distribution, and include reports from Italy 3 , Portugal 4 , the United States 5 . In Brazil, the fi rst occurrence of Leishmania infantum in a cat was recorded in 2004 by Savani et al 5 . Some authors consider this animal an accidental host, while others suggest that felids can act as potential peridomestic reservoirs 6 . However, the role of cats in the epidemiology of leishmaniasis remains to be clarifi ed.The present study was approved by the Animal Ethics Committee (protocol 65/2007) of the School of Veterinary Medicine and Animal Husbandry, São Paulo State University, following the Ethical Principles in Animal Experimentation. A total of 50 cats (Felis catus) that were domiciliated in the City of Campo Grande (20°26'34''S, 54°38'47''W) in the State of Mato Grosso do Sul, Brazil were selected randomly without predilection for sex breed, or age. Blood samples were obtained to detect antibodies for T. gondii and Leishmania spp.The immunofl uorescence antibody test (IFAT) was performed, according to the method described by Camargo 7 , which used whole Leishmania major-like promastigote and the RH strain of T. gondii as the antigen for the diagnosis of leishmaniasis and toxoplasmosis, respectively. Modifi ed agglutination test (MAT) using the RH strain was performed, as described by Desmonts and Remington 8 . The cutoff T. gondii antibody titer for both tests was 16, and the corresponding value in IFAT for Leishmania spp. was 40.Two (4%) of the 50 animals presented titers of antibodies for T. gondii and Leishmania spp., which suggested coinfection by these two protozoa in the same animal. Cat #1 was a female of undefi ned breed, 2 years of age, and presented T. gondii antibody titers of 64 (based on IFAT) and 1,024 (based on MAT), and an antibody titer of 320 for Leishmania spp. Cat #2 was also a female of undefi ned breed, 3 years of age, and presented T. gondii titers of 16 (based on IFAT) and 1,024 (based on MAT), and a Leishmania spp. titer of 320 (Table 1). Neither cat presented clinical signs. Similar results were obtained by Cardia et al. 9 with specifi c anti-Toxoplasma immunoglobulin G (IgG) in 63 (16.3%) of 386 cats and IgG against Leishmania spp. in two serum samples with titers of 1:160 and 1:320, respectively. In a study conducted by Sobrinho et al. 10 in another Brazilian endemic area, 25.7% cats were coinfected with both agents. However, the authors did not observe a signifi cant association between these infections.In this study, the MAT and IFAT were used, based on the results for cats obtained by Macrì et al. 11 , with a concordance of 0.98 (i.e...