Tegumentary and visceral leishmaniasis are severe and unfortunately common parasitic diseases in Brazil. Among domestic animals, dogs are considered the main urban reservoir of the protozoan parasites, however, there is evidence that infected cats can also contribute towards the disease pool. The number of cats diagnosed with leishmaniasis has greatly increased in the last few years, highlighting the importance of thorough investigations on the role of the cat in the epidemiological cycle of the disease and in public health related issues. The main clinical manifestations of leishmaniasis suffered by cats, even when infected with Leishmania chagasi, a viscerotropic species, are skin abnormalities, which can be confounded with multiple other diseases. Indirect ELISA should be used as a screening test in epidemiological investigations for being a sensitive technique, followed by more specific laboratory tests. The standardization and validation of rapid, economical and reproducible diagnostic methods, to be employed in epidemiological surveillance, are still required. Key words: Leishmania spp., cat, epidemiology, diagnosis, zoonosis, prevalence ResumoLeishmaniose tegumentar e visceral são doenças parasitárias graves e, infelizmente, comuns no Brasil. Entre os animais domésticos, o cão é considerado o principal reservatório urbano do parasito protozoário; no entanto, há indícios de que gatos infectados também possam contribuir para essas doenças. O número de gatos com diagnóstico de leishmaniose aumentou muito nos últimos anos, destacando a importância de investigações aprofundadas sobre o papel desse hospedeiro no ciclo epidemiológico da doença e em Saúde Pública. As principais manifestações clínicas da leishmaniose felina são anormalidades na pele, o que pode ser confundida com várias outras doenças, mesmo em casos de infecção por Leishmania chagasi, uma espécie viscerotrópica. ELISA indireto poderia ser usado como teste de triagem em investigações epidemiológicas, por ser um método sensível, seguido de exames laboratoriais mais específicos. A padronização e validação de métodos de diagnóstico rápidos, econômicos e reprodutíveis a serem empregados na vigilância epidemiológica ainda são necessárias.

The aim of this study was to detect cross infections by Leishmania spp. and Trypanosoma spp. using enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibody test (IFAT) and polymerase chain reaction (PCR). Thus, 408 blood samples were collected from dogs domiciled in Araçatuba Municipality, São Paulo State, Brazil; the dogs were of both sexes, of several breeds and aged 6 months. For Leishmania spp., 14.95% (61 out of 408) of dogs were reactive using IFAT. Positivity was 20.10% (82 out of 408) using ELISA and 29.66% (121 out of 408) using PCR, with significant differences for the sex and age of these animals (p < 0.05). For Trypanosoma spp., antibody occurrence using ELISA was 10.54% (43 out of 408), while PCR indicated 2.45% (10 out of 408) positive dogs. Using IFAT, 10.29% (42 out of 408) of animals were considered positive and only sex showed a significant difference (p < 0.05). In this study, 10.54% (43 out of 408) of animals were seropositive according to ELISA for Trypanosoma spp., of which 79.07% (34 out of 43) showed positive results in the molecular diagnosis for Leishmania spp., while of the 10.29% (42 out of 408) positive dogs according to IFAT, 95.24 % (40 out of 42) had confirmed infection by this parasite. The obtained results demonstrate evidence of cross infections by both protozoa in the animals analysed in this study.

With the aim of determining the occurrence of Cryptosporidium spp., 222 fecal samples were collected from Murrah buffalo calves aged up to 6 mo. Fecal DNA was genotyped with a nested polymerase chain reaction targeting the 18S rRNA gene and sequencing of the amplified fragment. Nested 18S PCR was positive for 48.2% of the samples. Sequence analysis showed that the most frequent species in these animals was Cryptosporidium ryanae, which was present in buffalo calves as young as 5 d. The zoonotic species Cryptosporidium parvum was detected in one animal. An uncommon Cryptosporidium 18S genotype was found in buffaloes.

ResumoEstudos referentes á criptosporidiose são essenciais, devido sua relevância em saúde pública e por sua patogenicidade em animais de produção e de companhia. Nos últimos 20 anos, houve uma rápida expansão das pesquisas envolvendo o gênero Cryptosporidium, em grande parte relacionada a estudos moleculares, propiciando a descrição de várias espécies, genótipos e subtipos do parasito. A caracterização molecular de isolados de diferentes origens (animal, humana e ambiental) tem sido amplamente usada com o intuito de investigar o potencial zoonótico deste protozoário. As formas de transmissão documentadas são de animais para o ser humano, de pessoa para pessoa, por meio de ingestão ou do uso de água destinada a lazer contaminada direta ou indiretamente com oocistos esporulados. A elevada taxa de animais naturalmente infectados e a susceptibilidade ao protozoário justificam a importância de se atentar para a ocorrência desta enfermidade. Assim, são demonstrados aspectos epidemiológicos desta zoonose em animais domésticos. Palavras-chave: Cryptosporidium spp., ruminantes, equinos, caninos, felinos AbstractStudies related cryptosporidiosis will be essential, due to its relevance in public health and pathogenicity in pets and production animals. Over the past 20 years, there has been a rapid expansion of research involving the Cryptosporidium genus, largely related to molecular studies, providing a description of

Sand flies are recognized as the major vector of canine visceral leishmaniasis. However, in some areas of Brazil where sand flies do not occur, this disease is found in humans and dogs. There has been speculation that ticks might play a role in transmission of canine visceral leishmaniasis and the DNA of Leishmania spp. has been reported in whole ticks. We investigated the presence of Leishmania spp. promastigotes in the intestines, ovaries, and salivary glands of Rhipicephalus sanguineus ticks collected from tick-infested dogs in two cities of Brazil. We used 66 dogs that tested positive and 33 that tested negative for Leishmania spp. according to direct cytological examination assays. Ten ticks were collected from each dog and dissected to collect the intestines, ovaries, and salivary glands for immunohistochemistry (IHC) and diagnostic real-time polymerase chain reaction (RT-PCR). IHC results showed Leishmania spp. in 98, 14, and 8 % of the intestines, ovaries, and salivary glands, respectively. Real-time PCR showed that 89, 41, and 33 % of the tick intestine, ovary, and salivary glands, respectively, were positive for Leishmania spp. The verification of promastigotes of Leishmania spp. by two independent techniques in ticks collected from these urban region dogs showed that there is need for clarification of the role of ticks in the transmission of canine visceral leishmaniasis in Brazil.