An epidemiological Leishmania spp. and entomological Phlebotomine sandflies survey was performed in cat shelters at leishmaniasis endemic area of Brazil. Blood and conjunctival swab (CS) samples were collected from 94 cats in two animal protection shelters. These samples were subjected to serological tests using the indirect immunofluorescence antibody test (IFAT) and indirect enzyme‐linked immunosorbent assay (ELISA) and to molecular test by polymerase chain reaction (PCR). In addition, a Phlebotomine sandflies survey was performed in the same shelters. The analyses revealed a positivity of 31.91% (30/94) through ELISA and 29.79% (28/94) through IFAT. The two serological tests showed a positive association with perfect agreement (k = 0.925). None of the cats were positive by Leishmania spp. DNA. One Lutzomyia (Lutzomyia) longipalpis male was found in one of the cat shelters. The results and the implications of our findings are discussed below.
Leishmaniasis is a neglected tropical disease that continues to pose a serious public health problem. Albeit dogs have long been held as the major reservoirs of Leishmania infantum, the involvement of domestic cats in the zoonotic cycle of visceral leishmaniasis has gained prominence. Here, 240 cats were evaluated by clinical signs and haematological/biochemical changes compatible with leishmaniasis and were diagnosed by serological, molecular, and parasitological techniques. Thus, four cats naturally infected by L. infantum were submitted to xenodiagnosis. A total of 203 females of Lutzomyia longipalpis were subjected to feeding on four cats, with all females completing the blood meal. Parasitological and molecular assays were carried out to evaluate the presence of L. infantum in the sand flies’ midgut. Promastigotes were observed in 10 females (6.5%) that fed on one cat, and L. infantum DNA was detected in 17 (8.4%) females that fed on two cats. Our results strengthen the evidence that naturally infected cats are capable of transmitting L. infantum to sand flies.
The aim of this study was to compare molecular tests used to diagnose Leishmania spp. in dogs with different stages of infection. Blood and conjunctival swab (CS) samples from dogs classified in four clinical stages were subjected to different PCR protocols (13A/13B, MC1/MC2, LITSR/L5.8S and LEISH-1/LEISH-2 primers). To the study, 22.3% (48/215) of dogs were classified as without clinical signs, 67.5% (145/215) stage I (mild disease), 7.0% (15/215) stage II (moderate disease) and 3.2% (7/215) stage III (severe disease). The results showed that in blood samples, 13A/13B detected a significant higher number of positive dogs in stage I (25/145) and in total (42/215) (p≤0.05). However, when CS samples were tested, no difference was observed (p>0.05). On the other hand, in blood samples, MC1/MC2 detected significantly fewer positive dogs classified as without clinical signs (0/48), in stage I (0/145) and in total (1/215) (p≤0.05). Likewise, in CS samples, this primers showed also lower detection (1/215) (p≤0.05). So than, we can conclude that PCR on blood samples with 13A/13B primers has greater capacity to detect positive dogs, mainly at the initial of clinical disease than do other primers and MC1/MC2 are not a good choice to detect Leishmania infantum infection in dogs.
Leishmania spp. are important agents of human and animal leishmaniases that have an important impact on public health. In this study, we aimed to detect the circulation of Leishmania spp. in cattle from a visceral leishmaniasis non-endemic area of the state of São Paulo, Brazil. DNA was extracted from blood samples from 100 heifers in the municipality of Pirassununga and was amplified using primers specific for the first internal transcriber spacer (ITS1), to assess the presence of trypanosomatids. The assays revealed that one sample presented bands of between 300 and 350 base pairs. In GenBank, this sample matched 100% with Leishmania infantum (314 base pairs). The results suggest that cattle can be infected by Leishmania infantum in Brazil.
Introduction: Visceral leishmaniasis (VL) transmission has been associated with two different populations of the Lutzomyia longipalpis complex in São Paulo state. Methods: In a recent focus of VL, we captured and dissected sand flies and investigated Leishmania infantum infection by parasitological, PCR, and sequencing analysis. Results: Flagellates were observed in 2 of 47 (4.2%) cembrene-1 Lu. longipalpis females. The sequences obtained matched those of Le. infantum. Conclusions: We found that the transmission of Le. infantum by cembrene-1 females may occur at a high rate in this focus of VL and presented new data on the vector capacity of this population.
Many owners believe to act in their pets' best interest by allowing them to go outside, with or without supervision. However, outdoor exposure greatly increases the risk of disease and accidents. In this study, we evaluated the association between infectious and parasitic diseases and the outdoor habits of pet dogs and cats. Epidemiological data were obtained from the records of dogs and cats treated at the Teaching Clinic and Hospital Unit of Veterinary Medicine in the city of Pirassununga, state of São Paulo, Brazil. Pets with any form of infection were included in the case study group, and pets with no infectious or parasitic diseases were used as controls. Animals were further divided according to their habits into indoor animals, indoor animals taken for walks, and outdoor animals. The odds ratio of having a disease was calculated from the comparisons among these groups using the MedCalc Statistical Software. We found an increased risk for the occurrence of infectious or parasitic diseases in outdoor dogs and cats when compared to indoor animals (OR of 4.735) and to those taken for walks (OR of 2.303). In light of our results, we suggest that awareness campaigns should also focus on the benefits of keeping pets indoors. KEY-WORDS:Access to outdoor. Odds ratio. Infection. Canine. Feline. RESUMOMuitos proprietários acreditam agir no melhor interesse de seus animais de estimação permitindo-os o acesso ao ambiente externo, com ou sem supervisão. Entretanto, isso aumenta consideravelmente o risco para a ocorrência de doenças e acidentes. Neste estudo, foi avaliada a associação entre doenças infecciosas e parasitárias com relação aos hábitos semi-domiciliados de cães e gatos. Dados epidemiológicos foram obtidos de arquivo de prontuários de cães e gatos atendidos na Unidade Didática Clínico Hospitalar (UDCH) do curso de Medicina Veterinária da FZEA/USP no município de Pirassununga, estado de São Paulo, Brasil. Animais de estimação que apresentaram doenças infectoparasitárias foram incluídos no grupo de casos; animais sem tais doenças, controles. Os animais foram ainda divididos de acordo em domiciliados, domiciliados frequentemente levados a passeios e semi-domiciliados. A razão de chances para a ocorrência de doenças infecciosas foi calculada a partir da comparação dos grupos supracitados, fazendo uso do software estatístico MedCalc. Encontrou-se um risco aumentado para a ocorrência de doenças infecto-parasitárias em cães e gatos semi-domiciliados quando comparados aos animais domiciliados (OR de 4.735) e quanto aos domiciliados guiados em passeios (OR de 2.303). À luz dos resultados se sugere que campanhas de conscientização foquem também nos benefícios da criação domiciliada de animais de estimação. PALAVRAS-CHAVE:Acesso à rua. Razão de chances. Infecçã o. Caninos. Felinos.
Atualmente, temos observado importantes mudanças nos padrões de transmissão da leishmaniose tegumentar (LT) e visceral (LV) em áreas endêmicas do Brasil. Além disso, a LV vem avançando rapidamente para novas áreas antes consideradas indenes. Os flebotomíneos (Diptera: Psychodidae: Phlebotominae) são pequenos dípteros com grande relevância em ciências médicas e veterinárias, por serem vetores de vírus, Bartonella bacilliformis e principalmente Leishmania spp. Os estudos sobre esses insetos são fundamentais para uma melhor compreensão da complexa epidemiologia das leishmanioses, revelando os aspectos biológicos desses vetores e suas relações com o parasita e seus reservatórios. Atualmente, abordagens moleculares vêm sendo incluídas nesses estudos e mostram-se ser ferramentas poderosas em pesquisas sobre esses dípteros. O objetivo desse trabalho, foi investigar os aspectos bioecológicos de flebotomíneos em área endêmica de LT e LV no estado de São Paulo, Brasil. Assim, foi realizado um levantamento entomológico para descrever a fauna, sazonalidade, taxa de PCR positivo para o DNA de Leishmania spp. e as fontes de repasto sanguíneo de flebotomíneos na área de estudo. Os insetos foram triados, identificados e o tórax e abdômen das fêmeas amostradas submetidas a extração de DNA e posterior PCR para a detecção do DNA do Kinetoplasto (kDNA) de parasitas do gênero Leishmania. Em seguida, amostras de DNA de fêmeas ingurgitadas, semi-ingurgitadas e grávidas foram submetidas a PCR para a amplificação do gene mitocondrial citocromo B (CYT-B) de vertebrados. Os produtos amplificados nas reações de CYT-B foram purificados e submetidos ao sequenciamento genético. As sequências encontradas foram confrontadas com a base de dados GenBank, para identificação da espécie de vertebrado envolvido no repasto sanguíneo, das fêmeas de flebotomíneos. Por empregar ferramentas moleculares, nesse trabalho, também foi analisado o desempenho de alguns métodos de extração de DNA em amostras individuais de tórax e abdômen de fêmeas desses insetos. Em nosso estudo, foram encontradas doze espécies de flebotomíneos, entre vetores comprovados e suspeitos nos ciclos epidemiológicos de LT e LV. Não foram observados padrões sazonais entre os fatores climáticos estudados (temperatura, umidade relativa do ar e precipitação) e a fauna desses insetos, capturados com armadilhas luminosas do tipo CDC. Sobre os métodos de extração de DNA, os métodos comerciais são uma boa opção para esse tipo de amostra. Contudo, os métodos caseiros baseado em fenol/clorofórmio/álcool isoamílico ou em NaCl/álcool apresentaram melhores resultados. As análises moleculares revelaram uma fêmea não ingurgitada de Ev. (Ald.) carmelinoi positiva na PCR do kDNA de Leishmania spp. A identificação das fontes de repasto sanguíneo revelou os suínos, humanos, cães, bovinos, galinhas e gambás como hospedeiros vertebrados de flebotomíneos na área em estudo. As implicações desses achados são aqui apresentadas na forma de dois artigos científicos submetidos para avaliação em revistas da área. P...
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