Introduction: In Brazil, visceral leishmaniasis (VL) has spread to various regions. This study reports canine cases of VL in Barra Mansa, where human VL cases were recently reported. Methods: Using the human index case, a canine survey was performed by dual-path platform immunochromatography and enzyme-linked immunosorbent assay. Seropositive animals were euthanized. Cultures were collected to detect Leishmania parasites. Results: Serological tests detected 141 canine VL cases, and Leishmania chagasi were isolated from 82.2% animals. Conclusions: Leishmania chagasi is in circulation in Barra Mansa. This study broadens information on the parasite's distribution in the State of Rio de Janeiro.

Visceral leishmaniasis caused by the protozoan Leishmania infantum is a zoonosis. The domestic dog is the primary reservoir in urban areas. This study aimed to evaluate the frequency, active infection and load of L . infantum in the genital tract of male and female dogs seropositive for this parasite, as well as to identify histological genital alterations associated with this protozoan. We studied 45 male and 25 female L . infantum -seropositive noncastrated dogs from the same endemic area in Brazil. Tissue samples from the testis, epididymis, prostate, vulva, vagina, and uterus were examined by singleplex qPCR and parasitological tests (histopathology, immunohistochemistry, and parasitological culture). The latter were performed for the detection of active infection (parasites able to multiply and to induce lesions). Forty-four (98%) males and 25 (100%) females were positive for L . infantum in the genital tract (epididymis: 98%; vulva: 92%; vagina: 92%; testis: 91%; uterus: 84%; prostate: 66%). Active infection in the genital tract was confirmed in 69% of males and 64% of females (32% in the uterus). Parasite loads were similar in the testis, vulva, epididymis and vagina and lower in the prostate. Only the parasite load in the vagina was significantly associated with the number of clinical signs. Granulomatous inflammation predominated in all organs, except for the prostate. Only in the testis and epididymis was the inflammatory infiltrate significantly more intense among dogs with a higher parasite load in these organs. The high frequency, detection of active infection and similarity of L . infantum loads in the genital tract of infected males and females suggest the potential of venereal transmission of this parasite by both sexes and of vertical transmission by females in the area studied. Additionally, vertical transmission may be frequent since active L . infantum infection was a common observation in the uterus.

Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffinembedded healthy skin for the diagnosis of canine visceral leishmaniasis

After the report of a second case of canine visceral leishmaniasis (CVL) in São Bento da Lagoa, Itaipuaçu, in the municipality of Maricá, Rio de Janeiro State, an epidemiological survey was carried out, through active search, totaling 145 dogs. Indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (ELISA), and rapid chromatographic immunoassay based on dual-path platform (DPP(r)) were used to perform the serological examinations. The parasitological diagnosis of cutaneous fragments was performed by parasitological culture, histopathology, and immunohistochemistry. In the serological assessment, 21 dogs were seropositive by IFA, 17 by ELISA, and 11 by DPP(r), with sensitivity of 66.7%, 66.7% and 50%, and specificity of 87.2%, 90.2% and 94%, respectively for each technique. The immunohistochemistry of bone marrow using the cell-block technique presented the best results, with six positive dogs found, three of which tested negative by the other parasitological techniques. Leishmania sp. was isolated by parasitological culture in three dogs. The detection of autochthonous Leishmania infantum in Itaipuaçu, and the high prevalence of seropositive dogs confirm the circulation of this parasite in the study area and alert for the risk of expansion in the State of Rio de Janeiro.

Introduction: Trypanosoma caninum is a protozoan species recently described in dogs, whose occurrence has been reported in areas of overlap with visceral leishmaniasis. Methods: Trypanosoma sp. were isolated from nine dogs and characterized by molecular methods. Results: PCR and sequencing confi rmed the presence of T. caninum in all dogs, revealing two new areas of transmission: Barra Mansa and São João do Piauí. Conclusions: The nine new cases described, when added to those already published, account for 62 cases of natural infection by T. caninum and show the geographical spread of this species to new areas, where other trypanosomatids circulate.

The objective of the current research was to assess seroconversion in dogs immunized with Leishmune® and Leish Tec® vaccines using rapid chromatographic immunoassay DPP® (Dual Path Platform) (DPP CVL) and enzyme immunoassay (EIE) up to one year after the vaccination protocol. The study sample comprised 28 dogs divided into two groups, each group immunized with an anti-CVL vaccine and clinically monitored for one year through clinical evaluation and laboratory tests. 22 (78.5%) dog were monitored. During the evaluation time (T1-30 days, T2-6 months, and T3-1 year after vaccination) the results for all dogs were negative for CVL, except for one animal vaccinated with Leish tec® that seroconverted in the DPP CVL test at T2. Subsequent examinations of this dog were negative. Our results showed that in a non-endemic area, even at different evaluation times, dogs vaccinated against CVL with Leishmune® or Leish tec® did not seroconvert in the serological protocol used by the Brazilian Ministry of Health (DPP/EIE).

ResumoA leishmaniose visceral (LV) é uma endemia em franca expansão geográfica. Este relato apresenta um caso de importação de LV canina para o município de Resende, estado do Rio de Janeiro, onde não havia até o momento registro de casos humanos ou caninos da parasitose. O animal era oriundo do estado de Minas Gerais, área endêmica de LV, e apresentou clínica compatível com o processo patológico em questão. O diagnóstico foi realizado por meio de avaliação sorológica com o teste rápido imunocromatográfico DPP ® (Dual Path Platform) e teste de ELISA, ambos com resultado positivo, além de cultura parasitológica em Meio NNN acrescido de meio Schneider a partir de punções de medula óssea e linfonodos, biópsias de pele íntegra, lesões cutâneas, linfonodo e baço, obtendo-se o isolamento de formas promastigotas compatíveis com Leishmania spp., com posterior caracterização de Leishmania (Leishmania) chagasi através de eletroforese de isoenzimas. O levantamento entomológico com utilização de armadilhas luminosas do tipo CDC, realizado no peridomicílio três vezes por semana durante três meses, não evidenciou a presença de flebotomíneos, e o inquérito sorológico canino, procedido em 144 animais ao redor da residência do caso positivo de LV, descartou novos casos da parasitose por meio do teste rápido imunocromatográfico DPP ® e/ou confirmação por ELISA. Os resultados e o histórico de deslocamento do animal caracterizam o caso como alóctone, expõem a fragilidade em relação ao controle de trânsito de animais e ressaltam a importância das ações de Vigilância para a parasitose em áreas indenes. Palavras-chave : diagnóstico, cão, Leishmania. AbstractThe visceral leishmaniasis (VL) is an endemic parasitosis in intense geographic spread. This report presents a case of canine VL importation for the city of Resende, state of Rio de Janeiro, where had not been reported cases of human or canine disease yet. The animal was from Minas Gerais state, endemic area for VL, and showed compatible clinical signs of this disease. The diagnosis was made by serology with DPP ® (DUAL PATH PLATFORM) immunochromatographic rapid test and ELISA, both with positive results, as well as by parasitological culture in NNN medium plus Schneider's medium. For parasitological culture, samples from puncture of bone marrow and lymph node and biopsies of intact skin, skin lesions, lymph node and spleen were examined and resulted in isolation of promastigotes characterized as Leishmania (Leishmania) chagasi by multilocus enzyme electrophoresis. The entomological survey, which was made at peridomicile using CDC light traps, three times a week for three months, did not find sandflies. Additionally, the canine serosurvey conducted in 144 animals around the residence with positive case of LV did not show new cases through rapid immunochromatographic test DPP ® and/or through confirmation by ELISA. The results and moving historical of the animal characterize the case as alochthonous, expose the fragility of movement control of animals and highlight the importance of...

Abstract. Although direct examination methods are important for diagnosing leishmaniasis, such methods are often neglected because of their low sensitivity relative to other techniques. Our study aimed to evaluate the performance of bone marrow (BM) thick smears and cytocentrifugation tests as alternatives to direct examination for diagnosing canine visceral leishmaniasis (CVL). Ninety-two dogs exhibiting leishmaniasis seroreactivity were evaluated. The animals were euthanized; and healthy skin, spleen, popliteal lymph node, and BM puncture samples were cultured. BM cultures were used as the reference standard. Of the 92 dogs studied, 85.9% exhibited positive cultures, and Leishmania infantum (synonym Leishmania chagasi) was confirmed in all positive culture cases. The sensitivity rates for cytocentrifugation as well as thin and thick smears were 47.1%, 52.8%, and 77%, respectively. However, no association between the dogs' clinical status and culture or direct examination results was found. To our knowledge, this was the first study to use thick smears and cytocentrifugation for diagnosing CVL. Our results indicate that BM thick smears have a good sensitivity and their use reduces the time required to read slides. Therefore, thick smears can provide a rapid and safe alternative to parasitological confirmation of seroreactive dogs.