This study aimed to investigate anti-Toxoplasma gondii antibodies and to isolate the parasite from the brains of horses processed at slaughterhouses in Brazil. We collected brain and blood samples from 398 horses of various ages, from six Brazilian states. Serum samples were evaluated by indirect fluorescent antibody test (IFAT cut-off titre ≥ 1:64), and brains were submitted to mouse bioassay. Among the 398 horses, positivity for T. gondii was identified in 46 (11.6%) by IFAT and in 14 (3.5%) by mouse bioassay. In 12 of those 14 bioassays, mice were positive only by IFAT (cut-off titre ≥ 1:16), T. gondii being isolated in the remaining two. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of 18S rDNA to differentiate among T. gondii, Neospora caninum, and Sarcocystis neurona, we found that two of the 14 brains were positive for T. gondii only. For genotyping of the two isolates and the PCR-positive brain, we performed PCR-RFLP based on 13 markers, and SAG2 all samples were Toxoplasma gondii type I. Collectively, IFAT of horse sera and mouse bioassay identified positivity in 60 (15%) of the samples. Our results show that some horses sent to slaughter in Brazil have been exposed to T. gondii.Keywords: Slaughterhouse, horses, Toxoplasma gondii, bioassay, indirect fluorescent antibody test, nested PCR-RFLP. ResumoO objetivo do estudo foi investigar anticorpos anti-Toxoplasma gondii e isolar o parasita do cérebro de equídeos abatidos em matadouros-frigoríficos no Brasil. Colheram-se amostras de 398 cérebros e sangue de equídeos machos e fêmeas de idades variadas, provenientes de seis estados brasileiros. As amostras de soro foram avaliadas pelo teste de imunofluorescência indireta (IFI) para T. gondii (ponto de corte ≥ 64), e os fragmentos de cérebros foram submetidos ao bioensaio em camundongos. Por meio da IFI, 46 (11,6%) equídeos foram soropositivos. Pelo bioensaio em camundongos, 14 (3,5%) cérebros de equídeos testados foram positivos. Em doze dos bioensaios, os camundongos foram positivos somente pela IFI (ponto de corte ≥ 16) e T. gondii foi isolado nos outros dois bioensaios. A PCR-RFLP com base em 18S rDNA para diferenciar entre T. gondii, Neospora caninum, e Sarcocystis neurona foram feitas em todos os 14 cérebros e dois foram positivos apenas para T. gondii. De dois isolados positivos para T. gondii e do cérebro positivo à PCR em que realizou-se a PCR-RFLP, com base em 13 marcadores e SAG2, a genotipagem mostrou ser o T. gondii tipo I para todas as amostras. A IFI de soros de equídeos e do bioensaio em camundongos identificaram positividade em 60 (15%) amostras testadas. Os resultados mostram que alguns cavalos enviados para abate foram expostos ao T. gondii.
Toxoplasma gondii is a protozoan that has great genetic diversity and is prevalent worldwide. In 2018, an outbreak of toxoplasmosis occurred in Santa Maria, Brazil, which was considered the largest outbreak ever described in the world. This paper describes the isolation and molecular characterization of Toxoplasma gondii from the placenta of two pregnant women with acute toxoplasmosis who had live births and were receiving treatment for toxoplasmosis during the outbreak. For this, placental tissue samples from two patients underwent isolation by mice bioassay, conventional PCR and genotyping using PCR-RFLP with twelve markers. Both samples were positive in isolation in mice. The isolate was lethal to mice, suggesting high virulence. In addition, the samples were positive in conventional PCR and isolates submitted to PCR-RFLP genotyping presented an atypical genotype, which had never been described before. This research contributes to the elucidation of this great outbreak in Brazil.
Toxoplasmosis is a worldwide zoonosis with important public health implications. The aims of this study were to determine and identify factors associated to the seroprevalence of anti-Toxoplasma gondii antibodies in humans and dogs from Jataizinho, Parana State, Brazil. Serum samples from 280 humans and 766 dogs, from rural and urban areas, were tested by indirect immunofluorescent antibody test. Positive dilutions were ≥ 1:16. Epidemiological informations were obtained through an individual questionnaire and data tabulation was performed using EpiInfo 3.5.3 (CDC, Atlanta). Statistical significance was evaluated by chi-square (ᵪ 2 ) or Fisher's exact tests with a 5.0 % significance level. The seroprevalence in humans was 70.00% (196/280), as follows: 69.08% (143/207) in rural and 72.60% (53/73) in a slum urban community. The seroprevalence in dogs was 52.35% (401/766), as follows: 71.74% (99/138) in rural and 48.08% (302/628) in urban area. In rural areas, there was no factor associated to risk of infection. In urban slum community, visualizing more than four rats at home was a factor statistically associated to seropositivity for human toxoplamosis. Infection of urban dogs was associated to age, contact with stray dogs or rodents, and free access to street. The results indicated a similar possibility of contact with Toxoplasma gondii between dogs and humans living in rural or slum urban areas of small municipalities.Keywords: IFAT; prevalence; Toxoplasma gondii; zoonosis. ResumoA toxoplasmose é uma doença de distribuição mundial com importantes implicações em saúde 2 Cienc. anim. bras., Goiânia, v.18, 1-9, e-42102, 2017Seroepidemiology of toxoplasmosis in humans and dogs… pública. Os objetivos deste estudo foram determinar e identificar fatores associados à soroprevalência de anticorpos anti-Toxoplasma gondii em humanos e cães de Jataizinho, Paraná, Brasil. Amostras de soro de 280 humanos e 766 cães, de áreas rural e urbana, foram testadas pela reação de imunofluorescência indireta (RIFI). Foram consideradas positivas diluições ≥1:16. Informações epidemiológicas foram obtidas por meio de questionário individual e a tabulação dos dados foi realizada com EpiInfo 3.5.3 (CDC, Atlanta). A significância estatística foi avaliada por Chi-quadrado (ᵪ 2 ) ou Exato de Fisher com nível de significância de 5,0 %. A soroprevalência em humanos foi como segue: 70,00% (196/280), 69,08% (143/207) em área rural e 72,60% (53/73) em uma comunidade urbana carente. Em cães, foi 52,35% (401/766), na área rural 71,74% (99/138) e na área urbana 48,08% (302/628). Nas áreas rurais não houve fator estatisticamente associado ao risco de toxoplasmose. Na área urbana, visualizar mais do que quatro ratos em casa foi fator associado à soropositividade para toxoplasmose em humanos. A soropositividade em cães urbanos apresentou associação à idade, ao contato com cães errantes ou roedores e com acesso livre à rua. Os resultados indicaram semelhante possibilidade de contato com Toxoplasma gondii entre cães e humanos residentes em áreas rura...
The objective of this study was to report an outbreak of human toxoplasmosis at a research institution in Londrina, Paraná, from December 2015 to February 2016. Blood samples from 26 symptomatic individuals were collected and the microparticle chemiluminescence immunoassay was performed to detect IgM, IgG and specific IgG avidity test in the official laboratory. A total of 20 people with symptoms and serology compatible with acute toxoplasmosis (IgM positive and IgG with low avidity) were selected as cases, while 45 asymptomatic employees working in the same teams and during the same shifts were selected as controls. All the participants of the investigation answered an epidemiological questionnaire. Three samples of water and one sludge from the institution's supply cisterns, 10 soil samples, 11 plant samples, three cat fecal samples and one domestic feline cadaver were collected for analysis of the polymerase chain reaction (PCR) for T. gondii. After analyzing the epidemiological data, the consumption of vegetables in the restaurant of the institution was the only variable associated with the occurrence of the disease. In laboratory results, all the samples showed negative results to PCR. The rapid recognition of the outbreak, early notification and investigation could have broken the chain of transmission early, thus preventing the emergence of new cases. In addition, the adoption of good food handling practices could have prevented the occurrence of the outbreak.
The objective of this study was to investigate the presence of anti-Leishmania spp. antibodies using an indirect immunofluorescence technique in the serum of equidae slaughtered in two slaughterhouses registered with the Federal Inspection Service. Titers greater than 40 were considered reagents. Blood samples were collected from 398 animals of both sexes, with different ages, and coming from six Brazilian states, of which 46.0% (183/398) were reactive, indicating that these animals were exposed to the leishmaniasis agent that is widely distributed in Brazil. Key words: Slaughterhouse. Equine. Leishmaniosis. IFAT. Serology. ResumoO objetivo deste estudo foi investigar a presença de anticorpos anti-Leishmania spp. utilizando a técnica de imunofluorescência indireta em amostras de soro de equídeos abatidos em dois matadouros registrados no Serviço de Inspeção Federal. Os títulos superiores a 40 foram considerados reagentes. Foram coletadas amostras de sangue de 398 animais de ambos os sexos, com diferentes idades e provenientes de seis estados brasileiros, dos quais 46,0% (183/398) foram reativos, indicando que esses animais foram expostos ao agente de leishmaniose, amplamente distribuído no Brasil. Palavras-chave: Matadouro. Equinos. Leishmaniose. RIFI. Sorologia.
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