The aim of the present study was to evaluate the serological methods using ELISA with recombinant-rK39 (ELISA-rK-39) and soluble extract-SE (ELISA-SE) antigens, the indirect fluorescence antibody test (IFAT) in comparison to an immunochromatography rapid diagnostic test (RDT-rK39) and with a direct parasitological exam (PA) for Canine Visceral Leishmaniasis (CVL) diagnosis. The results showed that 89% (60/67) of the dogs were positive for at least one serological diagnostic test. ELISA-SE was the test that detected anti-Leishmania antibodies in the serum of the highest number of dogs (71.6%) followed by ELISA-rK39 (65.7%), IFAT (65.7%) and RDT-rK39 (55.2%). PA detected the lowest numbers (40.3%) of positive dogs. In relation to the total of examined dogs, the Kappa indexes (p ≤ 0.05) showed a good agreement between ELISA-SE and IFAT (88.1%; k = 0.7237), and it was also observed in the comparison of RDT-rK39 with ELISA-SE (83.6%, k= 0.6561), IFAT (83.5%, k= 0.6605) and PA (85.0%, k= 0.7074). A bad agreement was detected in any association of ELISA-rk39 with the other tests in either symptomatic or asymptomatic animals. ELISA as well as RDT using recombinant antigenic protein (rK39) were the methods that detected the lowest prevalence rates (33.3%) of CVL in asymptomatic dogs. In conclusion, only one test does not adequately identify dogs with CVL and it is necessary the association of two or more diagnostic tests. Because of the good agreement indexes of RDT-rK39 when evaluated with ELISA-SE, IFAT and PA it was suggested as a complementary method to be used in association with either ELISA-SE or IFAT, particularly in the symptomatic dogs. Furthermore, new studies are recommended in order to improve the sensitivity of tests mainly for asymptomatic dogs.Keywords: CVL, diagnosis, dogs, Leishmania infantum, rK39, RDT. ResumoO objetivo do presente estudo foi avaliar os métodos sorológicos usando ELISA (Ensaio Imunoenzimático Indireto) com o antígeno recombinante rK39 (ELISA-rK39) e o antígeno extrato solúvel bruto (ELISA-ES) e a RIFI (Reação de Imunofluorescência Indireta) em comparação com o método imunocromatográfico rápido (RDT-rK39) e o parasitológico direto (PA), para o diagnóstico da Leishmaniose Visceral Canina (LVC) em cães de Ilha Solteira, São Paulo, Brasil. Os resultados mostraram que 89% (60/67) dos cães foram positivos por pelo menos um teste diagnóstico sorológico (RIFI, ELISA-ES, ELISA-rk39 ou RDT-rK39) e somente 40,3% (27/67) foram positivos pelo PA. O ELISA-ES foi o teste que detectou anticorpos anti-Leishmania em maior número de cães (71,6%) seguido por ELISArK39, RIFI (65,7%) e por 2%). No total de cães analisados (assintomáticos e sintomáticos), o índice Kappa de concordância (p ≤ 0,05) foi considerado de boa concordância entre ELISA-ES e IFAT (88,1%; k= 0,7237) e entre RDT-rK39 com ELISA-ES (83,6%, k= 0,6561), RIFI (83,5%, k= 0,6605) e PA (85,0%, k= 0,7074). O índice de concordância ruim foi observado em qualquer associação de ELISA-rk39 com todos os outros testes nos animais sintomáticos ...
To elucidate the occurrence and epidemiology of leptospirosis in rats cohabitating with forest animals, 13 rats were captured at seven locations of the Centre for the Conservation of Wild Fauna (CCWF) in Sao Paulo state, Brazil, and samples of their blood, liver, and kidneys were collected. The diagnostic techniques utilized were the microscopic agglutination test (MAT), polymerase chain reaction (PCR) for Leptospira spp., and cultures of rat kidneys and liver in Fletcher's medium. The MAT results showed that 13 (100%) of the samples were reactive to 12 serovars among the 29 Leptospira spp. tested, and the Australis and Tarassovi serovars were the most frequently identified serovars. To research the agent in fragments of the liver and kidney, 13 samples from each tissue were cultured in Fletcher's medium, and the results revealed seven positive samples (53.8%; three from the kidneys and four from the livers). The analysis of the blood samples by PCR for Leptospira spp. showed that six animals (46.1%) were positive, whereas the analysis of the organs (kidneys and liver) by PCR revealed that nine animals (69.2%) were positive, and the culture of the organs revealed four positive animals (30.8%). These results suggest that the presence of Leptospira spp. infection in rats at the study site and the knowledge of the serovars that exist in this environment are important for the epidemiological comprehension of the disease and for the identification of control measures that should be considered to reduce the risk of transmission of the disease through this animal reservoir.
Visceral leishmaniasis (VL) is a zoonotic disease with worldwide distribution. The crab-eating fox (Cerdocyon thous) is considered a wild reservoir of many zoonotical diseases, particularly VL. This study reported the presence of Leishmania infantum amastigotes in different organs of one captive C. thous found dead in a zoo. This animal was positive by the indirect fluorescence antibody test and had many clinical signs of VL. Intracellular amastigote forms of L. infantum were seen in neutrophils and macrophages in sample tissues from skin, lymph nodes (popliteal, submandibular, prescapular, and mesenteric), spleen, and liver. The numbers of positive cells and intracellular parasites were higher in macrophages than in neutrophils. In addition, polymerase chain reaction demonstrated extensive distribution of Leishmania DNA in C. thous tissues from multiple organs. The presence of intracellular amastigotes in neutrophils and macrophages as well as DNA of the parasite in tissues, specifically skin demonstrate that this crab-eating fox is an adequate host for L. infantum and reinforce the importance of VL for symptomatic wild canids kept in captivity in endemic areas.
A Leishmaniose Visceral (LV), doença negligenciada de caráter crônico e sistêmico, causada pelo parasita Leishmania spp. representa um problema de saúde pública, principalmente em países com população vulnerável, como o Brasil. Devido à urbanização de grandes centros, cães configuram um importante reservatório no ciclo urbano, precedendo casos humanos e contribuindo para a manutenção de casos de Leishmaniose Visceral Canina (LVC). Entretanto, poucos inquéritos sorológicos somados à analise espacial envolvendo a população humana e animal foram descritos no município de Três Lagoas. Diante disso, o objetivo do estudo foi identificar e descrever a ocorrência da infecção natural por Leishmania spp. na população canina domiciliada e humana de Três Lagoas – MS, Brasil, utilizando-se das técnicas sorológicas de Dual Path Platform (DPP), Enzyme Linked Immunosorbent Assay (ELISA) e Reação de Imunofluorescência Indireta (RIFI), bem como molecular para Leishmania spp. e análise por georreferenciamento. Para isso, foram coletadas 566 amostras sanguíneas de cães. Além disso, informações epidemiológicas adicionais foram coletadas do sistema de saúde do município, e posteriormente, a análise espacial foi realizada. Como resultados obteve-se 267 amostras reagentes ao teste DPP, 267 amostras reagentes ao ELISA e 97 à RIFI, com 95% de concordância entre os métodos DPP e ELISA. Das amostras submetidas à PCR, 16 foram positivas para Leishmania spp. O uso do georreferenciamento para LVC canina em Três Lagoas forneceu subsídios para identificar áreas com prioridade de intervenção para o controle da doença. A presença de flebotomíneos, associado à localização geográfica, bem como às condições ambientais na região estudada, foram favoráveis à sua disseminação, bem como à transmissão da doença aos cães e a humanos.
Toxoplasmosis is one of the most common zoonoses worldwide. It was initially described in rodents and rabbits. There are few data on the morbidity and mortality of this disease among Brazilian marsupial fauna, such as opossums. These animals are of great importance regarding the epidemiology of this disease, given that they are prey for felids and other carnivores. With the aim of ascertaining the serological response to Toxoplasma gondii among marsupials (Didelphis spp.), 38 animals that had been caught in 14 districts of the urban area of the municipality of Bauru, state of São Paulo, were evaluated. The modified agglutination test (MAT) showed that 26.3% (10/38) of the samples analyzed were seropositive. It can be suggested that the opossums' behavior and persistent proximity to human housing results in contact with cats and T. gondii infection, based on the frequency found in this study. This was the first study on the seroprevalence of T. gondii in opossums caught in the urban area of the municipality of Bauru, SP, and it highlights the need for environmental and health authorities of the municipality to monitor this zoonosis.Keywords: Didelphis. Toxoplasmosis. Zoonosis. T. gondii. ResumoA toxoplasmose é uma das zoonoses mais comuns no mundo, tendo sido descrita inicialmente em roedores e em coelhos. Todavia, poucos são os dados sobre morbidade e mortalidade da toxoplasmose nos marsupiais da fauna brasileira, como os gambás, sendo de grande importância na epidemiologia da doença, como presas para felídeos e outros carnívoros. Com o objetivo de verificar a resposta sorológica para Toxoplasma gondii em marsupiais (Didelphis spp.), foram avaliados 38 animais capturados em 14 regiões da área urbana do município de Bauru-SP. Foi encontrada uma frequência, de acordo com o teste de aglutinação modificada (MAT), de 26,3% (10/38) nas amostras analisadas. Pode-se sugerir que o comportamento dos gambás e sua permanência próxima a habitações humanas resultam em contato com gatos e infecção por T. gondii, tendo em vista a frequência encontrada neste estudo. Este é o primeiro estudo de soroprevalência de T. gondii em gambás capturados na área urbana do município de Bauru-SP, alertando-se para a necessidade do monitoramento desta zoonose pelas autoridades de vigilância ambiental e sanitária do município.Palavras-chave: Didelphis. Toxoplasmose. Zoonoses. T. gondii.
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