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A pitiose, doença granulomatosa de eqüinos causada pelo oomiceto Pythium insidiosum, tem como característica a evolução rápida seguida de morte dos animais. Estas mortes muitas vezes são causadas por diagnósticos errôneos ou demorados quando os doentes já não respondem ao tratamento. Este trabalho teve por objetivo a padronização do ensaio imunoenzimático indireto (ELISA) para diagnóstico sorológico de pitiose em eqüinos e coelhos, visando a diminuição de erros e de tempo necessário para o diagnóstico. Para o desenvolvimento e validação do teste foram utilizadas 72 amostras de soro de eqüinos saudáveis e 44 soros de eqüinos com pitiose confirmada. Os resultados da validação do ELISA para eqüinos foram: sensibilidade 97,72%, especificidade 90,27%, valor preditivo positivo 86%, valor preditivo negativo 98,4% e eficiência de 93,1%. Para coelhos, o teste foi padronizado com 48 amostras de soro de animais saudáveis e 24 amostras de coelhos imunizados com antígenos de P. insidiosum. Os resultados foram: sensibilidade 91,66%, especificidade 95,83%, valor preditivo positivo 91,66%, valor preditivo negativo 95,83% e eficiência de 94,44%. Os resultados deste trabalho demonstram que o ensaio imunoenzimático indireto é um método seguro e eficaz para o diagnóstico sorológico da pitiose.

BackgroundLeishmaniosis, zoonosis that produces significant public health impacts, is caused by Leishmania infantum. Canines are the main domestic reservoir and, besides humans, other species of mammals could be infected when living in endemic areas. In this study, we detected equine Leishmania infantum infections in a canine visceral leishmaniosis transmission area and evaluated the clinical, haematological, biochemical and oxidative stress disorders. This study was conducted in Uruguaiana, Rio Grande do Sul, south of Brazil. Peripheral blood samples were collected from 124 animals (98 horses and 26 dogs) of both genders and several breeds after they underwent general and dermatologic examinations.ResultsTwenty five Leishmania infantum infected animals (20.16%), 14 horses and 11 dogs were detected by PCR (Polymerase Chain Reaction) amplification of kinetoplast DNA regions with 96% homology to Leishmania infantum (GenBank Accession No. L 19877.1). The clinical and haematological alterations of infected equines were skin lesions, nodules, lymphadenopathy, decreased levels in red blood cells and haematocrit (p < 0.05) and increase in urea serum concentration (p < 0.05), while CVL presented a decrease in red blood cells counts (p < 0.05), increase in lymphocytes (p < 0.05), and decrease in neutrophil-lymphocyte ratio (p < 0.05). Oxidative stress markers of plasma protein carbonyl and plasma lipid peroxidation were not statistically significant (p > 0.05) in both species.ConclusionsTo our knowledge, this has been the first leishmaniosis equine survey performed in south of Brazil, caused by Leishmania infantum that were able to initially identify haematological and biochemical changes in the species, even in asymptomatic animals. We present evidence supporting those findings of haematological and biochemical changes could be related to infection. Surprisingly, the clinical manifestations of equine infection were similar to those found in canine visceral leishmaniosis. The equine population could be play an important role in the cycle of leishmaniosis in south Brazil and consequently indicates a great risk of public health. This evaluation of infected animals is important to establish the clinical and laboratory parameters involved in the disease progression.

Chronological histological alterations of Metarhizium anisopliae during interaction with the cattle tick Boophilus microplus were investigated by light and scanning electron microscopy. M. anisopliae invades B. microplus by a process which involves adhesion of conidia to the cuticle, conidia germination, formation of appressoria and penetration through the cuticle. Twenty-four hours post-infection conidia are adhered and germination starts on the surface of the tick. At this time, the conidia differentiate to form appressoria exerting mechanical pressure and trigger hydrolytic enzyme secretion leading to penetration. Massive penetration is observed 72 h post-inoculation, and after 96 h, the hyphae start to emerge from the cuticle surface to form conidia. The intense invasion of adjacent tissues by hyphae was observed by light microscopy, confirming the ability of M. anisopliae to produce significant morphological alterations in the cuticle, and its infective effectiveness in B. microplus.

The present study aimed to evaluate factors associated with canine visceral leishmaniasis (CVL) in areas with higher seroprevalence of it in Uruguaiana city, Rio Grande do Sul state, Brazil, during the year of 2012. A semi-structured questionnaire with socioeconomic and environmental questions was applied in Leishmania endemic regions of the city. The survey data were analyzed by multivariate statistics and the associations between them were presented as odds ratio within a 95% confidence limit. The relationships between the studied variables showed no statistically significant difference between the dwellings with positive and negative dogs. However, when analyzing the odds ratio, the presence of green areas larger than 10m² turned the house into an area of potential risk for CVL (OR= 2.53). There was no difference among the socioeconomic variables education and income, though the groups with lower education and income showed a higher seroprevalence of CVL. The city of Uruguaiana is already located within the CVL broadcasting area and is taken as an endemic region of the disease, with seropositive dogs in all neighborhoods. Thus, the determinants for an increased Leishmania sp. infection could not be indicated, even so, the maintenance of green areas around the households represented a risk factor for being a potential vector shelter. Key words: Neglected disease. Epidemiology. Environment. Socioeconomic factors. ResumoO objetivo da presente pesquisa foi avaliar potenciais fatores de risco associados à ocorrência de leishmaniose visceral canina (LVC) em áreas de maior prevalência no município de Uruguaiana/RS, no ano de 2012. Foram aplicados questionários semiestruturados com questões socioeconômicas e ambientais em bairros endêmicos para LVC no município de Uruguaiana. Os dados foram analisados por meio de análise multivariada e as associações entre as variáveis foram expressas em odds ratio com intervalos de confiança de 95%. As relações entre as variáveis estudadas não apresentaram diferença estatística significativa quando se comparou residências com cães positivos àquelas com cães negativos. Entretanto, a presença de área verde maior que 10m² apresentou-se como um potencial fator de risco (OR= 2,53) para LVC. Não houve diferença significativa entre as variáveis socioeconômicas, escolaridade e renda embora, nos grupos de menor escolaridade e de menor renda familiar, tenha havido