First molecular detection of Leishmania (Leishmania) infantum chagasi in a domestic cat (Felis catus) from an urban area in eastern Amazon

Background: Domestic cats have been implicated as accidental hosts of Leishmania sp. However, in recent years, the recurrent description of new cases in endemic and nonendemic areas draw attention to the potential epidemiological role of cats as reservoir hosts. Although dogs are considered urban reservoirs, cats could act as a secondary natural reservoirs in these areas. Thus, feline leishmaniasis has become an emerging disease in several countries worldwide. Case presentation: This study aimed to describe the first case of feline leishmaniasis in a stray animal that presented lesions compatible with the disease in Belém, Pará, Brazil, an important urban area in eastern Amazon. Serological tests for Leishmania infantum (ELISA and IFA) were nonreactive, whereas histopathological examination indicated infectious dermatitis caused by Leishmania spp. or Toxoplasma gondii. Cytopathological study of lesion aspirate confirmed the presence of Leishmania sp. amastigotes within macrophages. Finally, molecular analyses revealed that the feline infection was caused by Leishmania (Leishmania) infantum chagasi. Conclusion: To the best of the authors' knowledge, this study reports the first case of natural infection by Leishmania (Leishmania) infantum chagasi in a feline from eastern Amazon. These findings suggest domestic cats as potential secondary reservoir hosts of Leishmania spp. in Belém, which reinforces the importance of further epidemiological investigation of feline leishmaniasis, especially in urban areas with human cases.(AU)

Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients

Background: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at ­80o C for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at ­20o C for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at ­80o C for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at ­20o C.(AU)

In-house serological ELISA as a leishmaniosis diagnostic test: development and applications in canines from the western border of Brazil / ELISA "in house" para o diagnóstico de leishmaniose: desenvolvimento e aplicação em caninos da fronteira Oeste do Brasil

ABSTRACT: Leishmaniosis is a great public health problem affecting both humans and animals. The disease is caused by the protozoan Leishmania spp., which has a complex cycle involving a phlebotomine vector. The ELISA test (Enzyme-Linked Immunosorbent Assay) along with a chromatographic immunoassay was defined by the Brazil Health Ministry as the confirmatory screening protocol in 2011. Uruguaiana city is 630 km away from Porto Alegre, which makes it difficult to send samples and diagnose leishmaniasis, as well as receive quick results. In view of this, the present study evaluated an in-house indirect ELISA method compared to indirect immunofluorescence assay (IFA) and dual-path platform chromatographic immunoassay (DPP-BioManguinhos®) for the detection of an immune response to Leishmania spp. in canine species. The serological evaluation included 48 canines from the western border of Brazil (Uruguaiana and Barra do Quaraí city). Among the 48 canine samples tested, 18 were positive when using the ELISA technique, 19 were positive with IFA, and 17 were positive with rapid test DPP®. The ELISA technique showed a sensitivity/specificity of 83.3%/86.7% when compared to IFA and 100%/96.8% compared to DPP®. The present study showed a prevalence of 37.5%, demonstrating that the infection circulates in the studied population. It can be concluded that the ELISA technique was valuable for use in field conditions when performing screening tests in endemic areas.

RESUMO: A leishmaniose é um grande problema de saúde pública que afeta tanto humanos quanto animais. A doença é causada pelo protozoário Leishmania spp., que possui um ciclo complexo envolvendo um vetor flebotomíneo. O teste ELISA (Enzyme-Linked Immunosorbent Assay), juntamente com um imunoensaio cromatográfico, foi definido pelo Ministério da Saúde do Brasil como protocolo de triagem confirmatória em 2011. A cidade de Uruguaiana fica a 630 km de Porto Alegre, o que dificulta o envio de amostras e o diagnóstico da leishmaniose, além da demora para obter os resultados. Diante disso, o presente estudo teve como objetivo avaliar um método de ELISA indireto in-house comparado ao ensaio de imunofluorescência indireta (IFA) e imunoensaio cromatográfico de plataforma de dupla via (DPP-BioManguinhos®) para a detecção de uma resposta imune contra Leishmania spp. na espécie canina. A avaliação sorológica incluiu 48 caninos da fronteira oeste do Brasil (cidade de Uruguaiana e Barra do Quaraí). Das 48 amostras caninas testadas, 18 foram positivas na técnica de ELISA, 19 foram positivas com IFA e 17 foram positivas com o teste rápido DPP®. A técnica de ELISA apresentou sensibilidade/especificidade de 83,3%/86,7% quando comparada ao IFA e 100%/96,8% em relação ao DPP®. O presente estudo apresentou prevalência de 37,5%, demonstrando que a infecção circula na população estudada. Pode-se concluir que a técnica de ELISA foi valiosa para uso em condições de campo na realização de testes de triagem em áreas endêmicas.

Review on trypanosomiasis and their prevalence in some country on the Red Sea

Abstract Trypanosomiasis is a protozoan infection affecting both human and animals in almost all parts of the world. It can affect a very large range of domestic and wild hosts including camelids, equines, cattle, buffaloes, sheep, goats, pigs, dogs and other carnivores, deer, gazelles and elephants. This review paper was designed to address the effect of this economically important disease in countries on the Red Sea, especially in Egypt, Sudan, Somalia, and Saudi Arabia during the period 2010 to 2020. The prevalence of trypanosomiasis is different between these countries due to different types of diagnostic methods (Giemsa-stained blood smears, Hematocrit centrifugation, Serological test, and molecular analysis PCR) used and differential distribution of vector (Tse tse) flies. In current review, retrospective studies of published literature on distribution and prevalence of Trypanosoma evansi infection in the Red Sea Countries was conducted [Google Scholar and PubMed were used to retrieve the published literature from 2000-2020. A total of 77 published articles met the eligibility criteria and were reviewed. A total of 16 reports have been reported on the prevalence and distribution of Trypnosoma evansi infection in the Red Sea Countries have been from 2010-2020]. According to the published literature, we can say that trypanosomiasis in camels are more prevalent in Sudan than in other countries, followed by 17% and 51.78% in both clinical and non-clinical cases. Hence, the reliable diagnostic tests should be used for rapid treatment or control of the disease as if not treated appropriately in early-stage, can lead to death of the camels.

Resumo A tripanossomíase é uma infecção por protozoário que afeta humanos e animais em quase todas as partes do mundo. Pode afetar grande variedade de hospedeiros domésticos e selvagens, incluindo camelídeos, equinos, gado, búfalos, ovelhas, cabras, porcos, cães e outros carnívoros, veados, gazelas e elefantes. Este artigo de revisão foi elaborado para abordar o efeito dessa doença economicamente importante em países do mar Vermelho, especialmente Egito, Sudão, Somália e Arábia Saudita, durante o período de 2010 a 2020. A prevalência de tripanossomíase é diferente entre esses países devido a tipos distintos de métodos diagnósticos (esfregaços de sangue corados com Giemsa, centrifugação de hematócrito, teste sorológico e PCR de análise molecular) usados e distribuição diferencial de moscas vetoras (tsé-tsé). Na revisão atual, foram realizados estudos retrospectivos da literatura publicada sobre distribuição e prevalência da infecção por Trypanosoma evansi nos países do mar Vermelho [Google Scholar e PubMed foram usados para recuperar a literatura publicada de 2000 a 2020. Um total de 77 artigos publicados preencheu os critérios de elegibilidade e foi revisado. E há também 16 relatos sobre a prevalência e distribuição da infecção por Trypnosoma evansi nos países do mar Vermelho, de 2010 a 2020]. De acordo com a literatura publicada, podemos afirmar que a tripanossomíase em camelos é mais prevalente no Sudão do que em outros países, seguida por 17% e 51,78% em casos clínicos e não clínicos. Assim, os testes diagnósticos confiáveis devem ser utilizados para o tratamento rápido ou controle da doença, pois, se eles não forem tratados de forma adequada na fase inicial, isso pode levar à morte dos camelos.

Prevalence and association of mycoplasma infection in the development of coronary artery disease / Prevalência e associação de infecção por micoplasma no desenvolvimento de doença arterial coronariana

Abstract Coronary heart disease (CHD) has been associated with significant morbidity and mortality worldwide. Although remain controversial, several studies have demonstrated the association of M. pneumoniae infections with atherosclerosis. We evaluated the possible association of mycoplasma infections in patients diagnosed with atherosclerosis by ELISA and PCR methods. Atherosclerotic tissue samples and blood samples were collected for the detection of mycoplasma antibodies (IgA) by ELISA from the 97 patients with coronary artery disease (CAD). M. pneumoniae specific IgA, IgG and IgM were measured by using the Anti-M. pneumoniae IgA/IgG/IgM ELISA. Detection of M. pneumoniae targeting the P1 adhesion gene was performed by PCR Acute infection of M. pneumoniae was diagnosed in 43.3% (42) of patients by PCR. The M. pneumoniae specific antibodies were detected in 36.1% (35) of patients. Twenty-five (25.8%) cases had IgG antibodies, 15 (15.5%) cases had IgM antibodies, 3 (3.1%) cases had IgA antibodies, 10 (10.3%) cases had both IgM + IgG antibodies and 1 (1%) case of each had IgM + IgA and IgG + IgA antibodies. None of the cases was positive for all three antibodies. A Pearson correlation coefficient analysis revealed an excellent correlation between the PCR and the serological results (r=0.921, p<0.001). A majority (17, 40.5%) of the M. pneumoniae positive patients are within the 41-50 years of age group, followed by 10 (23.8%) patients in the age group of 61-70 years and 2 (4.8%) patients were >70 years of age. Our study reported an unusually higher prevalence of M. pneumoniae by serological tests (36.1%) and PCR (43.3%). Although the hypothesis of the association of M. pneumoniae and CAD is yet to be proven, the unusually high prevalence of M. pneumoniae in CAD patients indicates an association, if not, in the development of atherosclerosis.

Resumo A doença coronariana (DCC) tem sido associada a significativa morbidade e mortalidade em todo o mundo. Embora ainda sejam controversos, vários estudos têm demonstrado a associação de infecções por M. pneumoniae com aterosclerose. Avaliamos a possível associação de infecções por micoplasma em pacientes com diagnóstico de aterosclerose pelos métodos ELISA e PCR. Amostras de tecido aterosclerótico e amostras de sangue foram coletadas para a detecção de anticorpos contra micoplasma (IgA) por ELISA de 97 pacientes com doença arterial coronariana (DAC). IgA, IgG e IgM específicos para M. pneumoniae foram medidos usando o Anti-M. pneumoniae IgA / IgG / IgM ELISA. A detecção de M. pneumoniae visando o gene de adesão P1 foi realizada por PCR. A infecção aguda por M. pneumoniae foi diagnosticada em 43,3% (42) dos pacientes pela PCR. Os anticorpos específicos para M. pneumoniae foram detectados em 36,1% (35) dos pacientes. Vinte e cinco (25,8%) casos tinham anticorpos IgG, 15 (15,5%) casos tinham anticorpos IgM, 3 (3,1%) casos tinham anticorpos IgA, 10 (10,3%) casos tinham anticorpos IgM + IgG e 1 (1%) caso de cada um tinha anticorpos IgM + IgA e IgG + IgA. Nenhum dos casos foi positivo para os três anticorpos. A análise do coeficiente de correlação de Pearson revelou uma excelente correlação entre o PCR e os resultados sorológicos (r = 0,921, p < 0,001). A maioria (17, 40,5%) dos pacientes positivos para M. pneumoniae está na faixa etária de 41-50 anos, seguida por 10 (23,8%) pacientes na faixa etária de 61-70 anos e 2 (4,8%) pacientes tinham > 70 anos de idade. Nosso estudo relatou uma prevalência incomumente maior de M. pneumoniae por testes sorológicos (36,1%) e PCR (43,3%). Embora a hipótese da associação de M. pneumoniae e DAC ainda não tenha sido comprovada, a prevalência incomumente alta de M. pneumoniae em pacientes com DAC indica uma associação, se não, no desenvolvimento de aterosclerose.

Pathology, microbiology, and molecular evaluation of tissues from equids serologically positive for Burkholderia mallei in Midwestern Brazil / Avaliação patológica, microbiológica e molecular em amostras de tecidos de equídeos sorologicamente positivos para Burkholderia mallei no Centro-Oeste do Brasil

Glanders is a disease caused by the bacterium Burkholderia mallei that primarily affects horses, mules and donkeys. The disease can cause lesions in the skin, lungs and several other organs. However, it often manifests as an asymptomatic disease. In Brazil, serological tests of high sensitivity and specificity are used to assist in the detection of antibodies against B. mallei and to contribute to the control of the disease. However, due to the mandatory euthanasia of seroreactive animals, equids with positive serology for B. mallei and asymptomatic generated great conflicts between breeders, veterinarians and diagnostic laboratories. This study clarifies the limitations of complementary diagnostic tests for detecting B. mallei. It describes the clinical, morphological and laboratory findings in 24 equines from different municipalities in the Mato Grosso State, Brazil, which reacted to the complement fixation test and were positive in the western blotting test for glanders. Data and tissue samples were collected from 24 horses for histological, microbiological and molecular analysis. In 23 horses, no clinical signs, morphological alterations, microbiological isolation, or molecular detection would characterize B. mallei infection. On the other hand, samples from an asymptomatic horse without lesional alterations showed sequence amplification compatible with B. mallei in the PCR. Considering that the infection by B. mallei is subject to the application of animal sanitary defense measures and that, by international requirement and national legislation, the serological results are tools that should support the sanitation procedures for the error of the bacteria in the Mato Grosso State, Brazil.

Mormo é uma enfermidade causada pela bactéria Burkholderia mallei que acomete primariamente cavalos, mulas e burros. A doença pode causar lesões na pele, pulmões e em diversos outros órgãos, entretanto frequentemente manifesta-se como uma enfermidade assintomática. No Brasil são utilizados testes sorológicos de elevada sensibilidade e especificidade para auxiliar na detecção de anticorpos contra B. mallei e contribuir para controle da doença. Porém, devido à obrigatoriedade da eutanásia de animais sororeagentes, os equídeos com sorologia positiva para B. mallei e assintomáticos geraram grandes embates entre criadores, médicos-veterinários e laboratórios de diagnóstico. Este trabalho esclarece as limitações dos testes diagnósticos complementares para detecção de B. mallei e descreve os achados clínicos, morfológicos e de exames laboratoriais em 24 equídeos, procedentes de diferentes municípios do estado de Mato Grosso, Brasil, que reagiram ao teste de fixação de complemento e foram positivos no teste de "western blotting" para mormo. Foram colhidos dados e amostras de tecidos de 24 equídeos para análise histológica, microbiológica e molecular. Em 23 equídeos não existiam sinais clínicos, alterações morfológicas, isolamento microbiológico ou detecção molecular que caracterizassem infecção por B. mallei. Por outro lado, amostras de um cavalo assintomático e sem alterações lesionais apresentaram amplificação de sequência compatível com B. mallei na PCR. Considerando que a infecção por B. mallei é passível da aplicação de medidas de defesa sanitária animal e que por exigência internacional e da legislação nacional, os resultados sorológicos são ferramentas que devem amparar os procedimentos de saneamento para erradicação da bactéria no estado de Mato Grosso, Brasil.

In-house serological ELISA as a leishmaniosis diagnostic test: development and applications in canines from the western border of Brazil / ELISA "in house" para o diagnóstico de leishmaniose: desenvolvimento e aplicação em caninos da fronteira Oeste do Brasil

Leishmaniosis is a great public health problem affecting both humans and animals. The disease is caused by the protozoan Leishmania spp., which has a complex cycle involving a phlebotomine vector. The ELISA test (Enzyme-Linked Immunosorbent Assay) along with a chromatographic immunoassay was defined by the Brazil Health Ministry as the confirmatory screening protocol in 2011. Uruguaiana city is 630 km away from Porto Alegre, which makes it difficult to send samples and diagnose leishmaniasis, as well as receive quick results. In view of this, the present study evaluated an in-house indirect ELISA method compared to indirect immunofluorescence assay (IFA) and dual-path platform chromatographic immunoassay (DPP-BioManguinhos®) for the detection of an immune response to Leishmania spp. in canine species. The serological evaluation included 48 canines from the western border of Brazil (Uruguaiana and Barra do Quaraí city). Among the 48 canine samples tested, 18 were positive when using the ELISA technique, 19 were positive with IFA, and 17 were positive with rapid test DPP®. The ELISA technique showed a sensitivity/specificity of 83.3%/86.7% when compared to IFA and 100%/96.8% compared to DPP®. The present study showed a prevalence of 37.5%, demonstrating that the infection circulates in the studied population. It can be concluded that the ELISA technique was valuable for use in field conditions when performing screening tests in endemic areas.

A leishmaniose é um grande problema de saúde pública que afeta tanto humanos quanto animais. A doença é causada pelo protozoário Leishmania spp., que possui um ciclo complexo envolvendo um vetor flebotomíneo. O teste ELISA (Enzyme-Linked Immunosorbent Assay), juntamente com um imunoensaio cromatográfico, foi definido pelo Ministério da Saúde do Brasil como protocolo de triagem confirmatória em 2011. A cidade de Uruguaiana fica a 630 km de Porto Alegre, o que dificulta o envio de amostras e o diagnóstico da leishmaniose, além da demora para obter os resultados. Diante disso, o presente estudo teve como objetivo avaliar um método de ELISA indireto in-house comparado ao ensaio de imunofluorescência indireta (IFA) e imunoensaio cromatográfico de plataforma de dupla via (DPP-BioManguinhos®) para a detecção de uma resposta imune contra Leishmania spp. na espécie canina. A avaliação sorológica incluiu 48 caninos da fronteira oeste do Brasil (cidade de Uruguaiana e Barra do Quaraí). Das 48 amostras caninas testadas, 18 foram positivas na técnica de ELISA, 19 foram positivas com IFA e 17 foram positivas com o teste rápido DPP®. A técnica de ELISA apresentou sensibilidade/especificidade de 83,3%/86,7% quando comparada ao IFA e 100%/96,8% em relação ao DPP®. O presente estudo apresentou prevalência de 37,5%, demonstrando que a infecção circula na população estudada. Pode-se concluir que a técnica de ELISA foi valiosa para uso em condições de campo na realização de testes de triagem em áreas endêmicas.

Review on trypanosomiasis and their prevalence in some country on the Red Sea / Avaliação da tripanossomíase e sua prevalência em alguns países do mar Vermelho

Abstract Trypanosomiasis is a protozoan infection affecting both human and animals in almost all parts of the world. It can affect a very large range of domestic and wild hosts including camelids, equines, cattle, buffaloes, sheep, goats, pigs, dogs and other carnivores, deer, gazelles and elephants. This review paper was designed to address the effect of this economically important disease in countries on the Red Sea, especially in Egypt, Sudan, Somalia, and Saudi Arabia during the period 2010 to 2020. The prevalence of trypanosomiasis is different between these countries due to different types of diagnostic methods (Giemsa-stained blood smears, Hematocrit centrifugation, Serological test, and molecular analysis PCR) used and differential distribution of vector (Tse tse) flies. In current review, retrospective studies of published literature on distribution and prevalence of Trypanosoma evansi infection in the Red Sea Countries was conducted [Google Scholar and PubMed were used to retrieve the published literature from 2000-2020. A total of 77 published articles met the eligibility criteria and were reviewed. A total of 16 reports have been reported on the prevalence and distribution of Trypnosoma evansi infection in the Red Sea Countries have been from 2010-2020]. According to the published literature, we can say that trypanosomiasis in camels are more prevalent in Sudan than in other countries, followed by 17% and 51.78% in both clinical and non-clinical cases. Hence, the reliable diagnostic tests should be used for rapid treatment or control of the disease as if not treated appropriately in early-stage, can lead to death of the camels.

Resumo A tripanossomíase é uma infecção por protozoário que afeta humanos e animais em quase todas as partes do mundo. Pode afetar grande variedade de hospedeiros domésticos e selvagens, incluindo camelídeos, equinos, gado, búfalos, ovelhas, cabras, porcos, cães e outros carnívoros, veados, gazelas e elefantes. Este artigo de revisão foi elaborado para abordar o efeito dessa doença economicamente importante em países do mar Vermelho, especialmente Egito, Sudão, Somália e Arábia Saudita, durante o período de 2010 a 2020. A prevalência de tripanossomíase é diferente entre esses países devido a tipos distintos de métodos diagnósticos (esfregaços de sangue corados com Giemsa, centrifugação de hematócrito, teste sorológico e PCR de análise molecular) usados ​​e distribuição diferencial de moscas vetoras (tsé-tsé). Na revisão atual, foram realizados estudos retrospectivos da literatura publicada sobre distribuição e prevalência da infecção por Trypanosoma evansi nos países do mar Vermelho [Google Scholar e PubMed foram usados ​​para recuperar a literatura publicada de 2000 a 2020. Um total de 77 artigos publicados preencheu os critérios de elegibilidade e foi revisado. E há também 16 relatos sobre a prevalência e distribuição da infecção por Trypnosoma evansi nos países do mar Vermelho, de 2010 a 2020]. De acordo com a literatura publicada, podemos afirmar que a tripanossomíase em camelos é mais prevalente no Sudão do que em outros países, seguida por 17% e 51,78% em casos clínicos e não clínicos. Assim, os testes diagnósticos confiáveis ​​devem ser utilizados para o tratamento rápido ou controle da doença, pois, se eles não forem tratados de forma adequada na fase inicial, isso pode levar à morte dos camelos.

Sarcocystis spp. detection in cattle using different diagnostic methods / Detecção de Sarcocystis spp. em bovinos usando diferentes métodos de diagnóstico

Cattle are considered intermediate hosts of Sarcocystis, which can cause clinical signs and lower performance in the acute phase of infection. Sarcocystis spp. are usually not visible to the naked eye during the post mortem inspection. Moreover, fresh microscopic examination and transmission electron microscopy techniques are difficult to apply to large samples. Therefore, extensive studies on Sarcocystis infection in cattle using molecular and serological methods are required. Here, we investigated Sarcocystis spp. infection in cattle using fresh microscopic examination and polymerase chain reaction of myocardium samples and compared the results with the presence of antibodies against Sarcocystis spp. in corresponding serum samples detected using indirect fluorescent antibody test. Microscopic Sarcocystis were observed in 100% of the myocardial samples, and Sarcocystis DNA was present in 86% (43/50) of these samples. Antibodies against Sarcocystis spp. were detected in 96% (48/50) and 80% (40/50) of the serum samples at 1:25 and 1:200 dilutions, respectively. The three associated methods (fresh microscopic examination, PCR and serology) showed good sensitivity and detection for Sarcocystis spp. compared with fresh microscopic examination (only), and they may facilitate diagnosis in live animals on a large scale as well as monitoring of the herd status.

Os bovinos são considerados hospedeiros intermediários de Sarcocystis, podendo causar sinais clínicos e menor desempenho na fase aguda da infecção. Sarcocystis spp. geralmente não são visíveis a olho nu durante a inspeção post mortem. Além disso, o exame microscópico a fresco e as técnicas de microscopia eletrônica de transmissão são difíceis de aplicar a uma amostras de grande tamanho. Portanto, são necessários extensos estudos sobre a infecção por Sarcocystis em bovinos usando métodos moleculares e sorológicos. Aqui, investigamos a infecção de Sarcocystis spp. em bovinos por meio de exame microscópico a fresco e reação em cadeia da polimerase de amostras de miocárdio e comparado os resultados com a presença de anticorpos contra Sarcocystis spp. em amostras de soro correspondentes detectadas usando o teste de anticorpos fluorescentes indiretos. Sarcocistos microscópicos foram observados em 100% das amostras de miocárdio, e o DNA de Sarcocystis estava presente em 86% (43/50) dessas amostras. Anticorpos contra Sarcocystis spp. foram detectados em 96% (48/50) e 80% (40/50) das amostras de soro nas diluições 1:25 e 1:200, respectivamente. Os três métodos associados (exame microscópico a fresco, PCR e sorologia) mostraram boa sensibilidade e detecção para Sarcocystis spp. em comparação com o exame microscópico fresco (apenas) e podem facilitar o diagnóstico em animais vivos em larga escala, bem como o monitoramento do status do rebanho.

Review on trypanosomiasis and their prevalence in some country on the Red Sea / Avaliação da tripanossomíase e sua prevalência em alguns países do mar Vermelho

Trypanosomiasis is a protozoan infection affecting both human and animals in almost all parts of the world. It can affect a very large range of domestic and wild hosts including camelids, equines, cattle, buffaloes, sheep, goats, pigs, dogs and other carnivores, deer, gazelles and elephants. This review paper was designed to address the effect of this economically important disease in countries on the Red Sea, especially in Egypt, Sudan, Somalia, and Saudi Arabia during the period 2010 to 2020. The prevalence of trypanosomiasis is different between these countries due to different types of diagnostic methods (Giemsa-stained blood smears, Hematocrit centrifugation, Serological test, and molecular analysis PCR) used and differential distribution of vector (Tse tse) flies. In current review, retrospective studies of published literature on distribution and prevalence of Trypanosoma evansi infection in the Red Sea Countries was conducted [Google Scholar and PubMed were used to retrieve the published literature from 2000-2020. A total of 77 published articles met the eligibility criteria and were reviewed. A total of 16 reports have been reported on the prevalence and distribution of Trypnosoma evansi infection in the Red Sea Countries have been from 2010-2020]. According to the published literature, we can say that trypanosomiasis in camels are more prevalent in Sudan than in other countries, followed by 17% and 51.78% in both clinical and non-clinical cases. Hence, the reliable diagnostic tests should be used for rapid treatment or control of the disease as if not treated appropriately in early-stage, can lead to death of the camels.

A tripanossomíase é uma infecção por protozoário que afeta humanos e animais em quase todas as partes do mundo. Pode afetar grande variedade de hospedeiros domésticos e selvagens, incluindo camelídeos, equinos, gado, búfalos, ovelhas, cabras, porcos, cães e outros carnívoros, veados, gazelas e elefantes. Este artigo de revisão foi elaborado para abordar o efeito dessa doença economicamente importante em países do mar Vermelho, especialmente Egito, Sudão, Somália e Arábia Saudita, durante o período de 2010 a 2020. A prevalência de tripanossomíase é diferente entre esses países devido a tipos distintos de métodos diagnósticos (esfregaços de sangue corados com Giemsa, centrifugação de hematócrito, teste sorológico e PCR de análise molecular) usados e distribuição diferencial de moscas vetoras (tsé-tsé). Na revisão atual, foram realizados estudos retrospectivos da literatura publicada sobre distribuição e prevalência da infecção por Trypanosoma evansi nos países do mar Vermelho [Google Scholar e PubMed foram usados para recuperar a literatura publicada de 2000 a 2020. Um total de 77 artigos publicados preencheu os critérios de elegibilidade e foi revisado. E há também 16 relatos sobre a prevalência e distribuição da infecção por Trypnosoma evansi nos países do mar Vermelho, de 2010 a 2020]. De acordo com a literatura publicada, podemos afirmar que a tripanossomíase em camelos é mais prevalente no Sudão do que em outros países, seguida por 17% e 51,78% em casos clínicos e não clínicos. Assim, os testes diagnósticos confiáveis devem ser utilizados para o tratamento rápido ou controle da doença, pois, se eles não forem tratados de forma adequada na fase inicial, isso pode levar à morte dos camelos.

Review on trypanosomiasis and their prevalence in some country on the Red Sea / Avaliação da tripanossomíase e sua prevalência em alguns países do mar Vermelho

Trypanosomiasis is a protozoan infection affecting both human and animals in almost all parts of the world. It can affect a very large range of domestic and wild hosts including camelids, equines, cattle, buffaloes, sheep, goats, pigs, dogs and other carnivores, deer, gazelles and elephants. This review paper was designed to address the effect of this economically important disease in countries on the Red Sea, especially in Egypt, Sudan, Somalia, and Saudi Arabia during the period 2010 to 2020. The prevalence of trypanosomiasis is different between these countries due to different types of diagnostic methods (Giemsa-stained blood smears, Hematocrit centrifugation, Serological test, and molecular analysis PCR) used and differential distribution of vector (Tse tse) flies. In current review, retrospective studies of published literature on distribution and prevalence of Trypanosoma evansi infection in the Red Sea Countries was conducted [Google Scholar and PubMed were used to retrieve the published literature from 2000-2020. A total of 77 published articles met the eligibility criteria and were reviewed. A total of 16 reports have been reported on the prevalence and distribution of Trypnosoma evansi infection in the Red Sea Countries have been from 2010-2020]. According to the published literature, we can say that trypanosomiasis in camels are more prevalent in Sudan than in other countries, followed by 17% and 51.78% in both clinical and non-clinical cases. Hence, the reliable diagnostic tests should be used for rapid treatment or control of the disease as if not treated appropriately in early-stage, can lead to death of the camels.(AU)

A tripanossomíase é uma infecção por protozoário que afeta humanos e animais em quase todas as partes do mundo. Pode afetar grande variedade de hospedeiros domésticos e selvagens, incluindo camelídeos, equinos, gado, búfalos, ovelhas, cabras, porcos, cães e outros carnívoros, veados, gazelas e elefantes. Este artigo de revisão foi elaborado para abordar o efeito dessa doença economicamente importante em países do mar Vermelho, especialmente Egito, Sudão, Somália e Arábia Saudita, durante o período de 2010 a 2020. A prevalência de tripanossomíase é diferente entre esses países devido a tipos distintos de métodos diagnósticos (esfregaços de sangue corados com Giemsa, centrifugação de hematócrito, teste sorológico e PCR de análise molecular) usados e distribuição diferencial de moscas vetoras (tsé-tsé). Na revisão atual, foram realizados estudos retrospectivos da literatura publicada sobre distribuição e prevalência da infecção por Trypanosoma evansi nos países do mar Vermelho [Google Scholar e PubMed foram usados para recuperar a literatura publicada de 2000 a 2020. Um total de 77 artigos publicados preencheu os critérios de elegibilidade e foi revisado. E há também 16 relatos sobre a prevalência e distribuição da infecção por Trypnosoma evansi nos países do mar Vermelho, de 2010 a 2020]. De acordo com a literatura publicada, podemos afirmar que a tripanossomíase em camelos é mais prevalente no Sudão do que em outros países, seguida por 17% e 51,78% em casos clínicos e não clínicos. Assim, os testes diagnósticos confiáveis devem ser utilizados para o tratamento rápido ou controle da doença, pois, se eles não forem tratados de forma adequada na fase inicial, isso pode levar à morte dos camelos.(AU)

Ocorrência de Babesia sp, Ehrlichia canis e Hepatozoon canis em cães domiciliados, em dois municípios do estado do Espírito Santo - Brasil / Occurrence of Babesia sp, Ehrlichia canis and Hepatozoon canis in domiciled dogs in two municipalities of the state of Espírito Santo - Brazil / Ocurrencia de Babesia sp, Ehrlichia canis y Hepatozoon canis em perros domiciliados em dos municipios del estado de Espírito Santo - Brasil

As doenças transmitidas por carrapatos são afecções de grande importância na clínica médica de pequenos animais, devido à alta casuística e ampla distribuição vetorial no território brasileiro. Os principais agentes responsáveis pelas infecções em cães são Babesia sp., Ehrlichia canis e Hepatozoon canis. Os animais infectados são assintomáticos ou apresentam sinais clínicos inespecíficos, sendo necessário a utilização de testes diagnósticos para definição do agente etiológico, e diagnóstico seguro. O objetivo do presente estudo foi determinar a ocorrência desses micro-organismos em cães naturalmente infectados, domiciliados nos municípios de Vila Velha e Anchieta, Espírito Santo, utilizando diferentes testes de detecção: Reação em cadeia polimerase (PCR), sorologia para detecção de anticorpos anti Ehrlichia canis e pesquisa de hematozoários em esfregaço sanguíneo. Foram analisadas 65 amostras de sangue obtidas por venopunção de veia cefálica de cães. No teste de PCR, 4,62% dos animais foram positivos para Babesia vogeli e 1,54% para Ehrlichia canis sendo os resultados para Hepatozoon canis negativos. No teste sorológico para E. canis 90,77% dos animais foram positivos para a presença de anticorpos, e na pesquisa em lâminas de esfregaço sanguíneo 3,02% apresentavam outros hemoparasitas. Os resultados indicam a dispersão desses hemoparasitas na população canina da região de estudo, entretanto com baixa ocorrência. O teste de PCR demonstrou-se como o mais sensível no qual Babesia vogeli foi o agente mais observado.(AU)

Tick-borne diseases are diseases of great importance in the medical practice of small animals, due to the high casuistry and wide vectorial distribution in the Brazilian territory. The main agents responsible for infections in dogs are Babesia sp., Ehrlichia canis and Hepatozoon canis. Infected animals are asymptomatic or present nonspecific clinical signs, requiring the use of diagnostic tests to define the etiologic agent, and safe diagnosis. The objective of the present study was to determine the occurrence of these microorganisms in naturally infected dogs domiciled in the municipalities of Vila Velha and Anchieta, Espírito Santo, using different detection tests: polymerase chain reaction (PCR), serology to detect antibodies against Ehrlichia canis and research of hematozoa in blood smears. Sixty-five blood samples obtained by venipuncture of the cephalic vein of dogs were analyzed. In the PCR test, 4.62% of the animals were positive for Babesia vogeli and 1.54% for Ehrlichia canis, and the results for Hepatozoon canis were negative. In the serological test for E. canis, 90.77% of the animals were positive for the presence of antibodies, and in the research in blood smear slides, 3.02% presented other hemoparasites. The results indicate the dispersion of these hemoparasites in the canine population of the study region, however with low occurrence. The PCR test proved to be the most sensitive, in which Babesia vogeli was the most observed agent.(AU)

Las enfermedades transmitidas por garrapatas son enfermedades de gran importancia en la práctica médica de los pequeños animales, debido a la alta casuística y amplia distribución vectorial en el territorio brasileño. Los principales agentes responsables de las infecciones en los perros son Babesia sp., Ehrlichia canis y Hepatozoon canis. Los animales infectados son asintomáticos o presentan signos clínicos inespecíficos, siendo necesario el uso de pruebas diagnósticas para la definición del agente etiológico, y el diagnóstico seguro. El objetivo del presente estudio fue determinar la ocurrencia de estos microorganismos en perros infectados naturalmente, domiciliados en los municipios de Vila Velha y Anchieta, Espírito Santo, utilizando diferentes pruebas de detección: reacción en cadena de la polimerasa (PCR), serología para detectar anticuerpos anti Ehrlichia canis e investigación de hematozoos en frotis de sangre. Se analizaron sesenta y cinco muestras de sangre obtenidas por venopunción de la vena cefálica de los perros. En la prueba PCR, el 4,62% de los animales fueron positivos para Babesia vogeli y el 1,54% para Ehrlichia canis, y los resultados para Hepatozoon canis fueron negativos. En la prueba serológica para E. canis, el 90,77% de los animales fueron positivos a la presencia de anticuerpos, y en la investigación en láminas de frotis de sangre el 3,02% presentaron otros hemoparásitos. Los resultados indican la dispersión de estos hemoparásitos en la población canina de la región de estudio, aunque con una baja presencia. La prueba PCR resultó ser la más sensible, en la que Babesia vogeli fue el agente más observado.(AU)

Enzyme-linked immunosorbent assay (ELISA) for detection of IgY Anti-Borrelia anserina antibodies in Gallus gallus domesticus / Ensaio de imunoadsorção enzimático (ELISA) indireto para detecção de anticorpos IgY Anti-Borreliaanserina em Gallus gallus domesticus

This study aimed to promote the standardization of an indirect, enzyme-linked immunosorbent assay (ELISA) for the serological detection of B. anserina in Gallus gallusdomesticus. An aliquoted sera from vaccinated chicken with B. anserina antigen (GI), experimental infected chickens with B. anserina (GII) and rustic poultry rearing of G. gallus (GIII) were tested with in-house ELISA developed to detect serum antibodies against B. anserina in G. gallus domesticus. On average, the experimentally infected chickens became positive at 9 DPI a mean ± standard deviation (SD) ODI value of 163.11 ± 70.65. The highest observed Optical Density Index (ODI) was 372.54 ± 132.39, at 26 DPI, and the highest overall ODI value was 626.51. The vaccinated chickens became positive between 8 and 10 DPV, with an ODI of 245.59 at 10 DPV, with an overall maximum ODI of 543.13. A total of 108 blood samples were collected from poultry raised on rustic farms. Of the total samples collected, 58.33% (63/108) were considered positive for B. anserina. The maximum ODI found among these rustic chickens was 283.24. This stardardization provided a sensitivity and specificity of 100%.

Este estudo teve como objetivo promover a padronização de um ensaio imunoenzimático indireto (ELISA) para a detecção sorológica de Borrelia anserina em Gallus gallus domesticus. Um frango vacinado com antígeno de B. anserina (GI), frangos infectados experimentalmente com B. anserina (GII) e frangos criados de forma rústica (GIII) foram testados com ELISA indireto in house desenvolvido para a detecção sorológica contra B. anserina em G. gallus domesticus. Em média, os frangos infectados experimentalmente tornaram-se positivos aos 9º dia pós-inoculação (DPI), um valor do índice de densidade óptica (ODI) médio ± desvio padrão (SD) de 163,11 ± 70,65. O maior ODI observado foi 372,54 ± 132,39, em 26ºDPI, e o maior valor geral de ODI foi 626,51. Os frangos vacinados tornaram-se positivos entre 8º e 10° DPV, com um ODI de 245,59 a 10 DPV, com um ODI máximo geral de 543,13. Um total de 108 amostras de sangue foram coletadas de aves criadas em fazendas rústicas. Do total de amostras coletadas, 58,33% (63/108) foram consideradas positivas para B. anserina. O ODI máximo encontrado entre essas galinhas rústicas foi 283,24. Essa padronização proporcionou sensibilidade e especificidade de 100%.

Leptospira spp. of the urinary tract of female carrier goats in semi-arid conditions

Background: Leptospirosis is an important infectious disease in goat farming, with a worldwide distribution. It is usually transmitted by rodents and the genital route, may cause reproductive losses, negatively impacting goat farming. The diagnosis lies on serological, molecular and isolation techniques. Considering the importance of this disease for small ruminants, this work aimed to evaluate the serological, molecular findings and isolation of pathogenic leptospires in the urinary tract (kidney and bladder tissues) of goats. Materials, Methods & Results: Thirty-four adult goats were used for slaughter. Renal samples (n = 34), bladder (n = 34), were collected for isolation of the agent and molecular detection of Leptospira sp. and blood samples (n = 34) for serological testing. The polymerase chain reaction (PCR) was used as a molecular test and the microscopic serum agglutination test (MAT) was used as a serological test. Samples with DNA amplification were subjected to genetic sequencing. The presence of Leptospira DNA was found in the tissues of 8 (23.4%) goats, and of these, only 2 were positive in PCR and MAT. There was a slight agreement between the PCR and MAT techniques (k = 0.150; P = 0.436). In 6 (17.6%) samples of renal tissue and 2 (5.8%) bladder samples, Leptospira DNA was detected. The genes in a kidney tissue sample were sequenced and demonstrated 99% similarity to Leptospira interrogans. Anti-Leptospira sp. were detected in 6 (17.6%) of the animals tested. Discussion: Serology identified 3 predominant serogroups: Icterohaemorrhagiae, Tarassovi and Autumnalis, serogroups that are related to the presence of rodents that coexist in rural environments. Autumnalis has been reported in small ruminants, raising the hypothesis that goats are adapted, becoming chronic carriers and possible maintenance hosts. The frequency obtained (17.6%) may be the result of the mixed breed pattern and rustic characteristics inherent to the goat species. Given the characteristics of the semi-arid region, such as low rainfall and high solar incidence, it is essential to use an adapted methodology, with a lower cut-off point (1:50), as the serological titer is an established relationship between the animal species, the level of exposure throughout its evolution and the region studied. Molecular findings and bacterial isolation reveal the agent's ability to colonize the urinary tract of goats. These data show the importance that urine has in the epidemiological chain, being able to transmit the agent through direct contact with this product or through contamination of soil and water. There was no statistical agreement between the diagnostic techniques used in this study, in this case, an association between PCR and MAT is recommended to obtain data with high sensitivity and specificity. A bladder sample was sequenced and showed 99% similarity to Leptospira interrogans. In the semiarid region, the most common form of leptospirosis spread is through the sale of animals in business fairs for breeding, rearing or slaughter, as well as sharing the same property with several breeders. The introduction of chronic and asymptomatic carriers on the properties represents a serious risk for the spread of the disease. The results show the presence of Leptospira spp. in semi-arid goat herds, having as risk factors the presence of rodents and intercropping. The association of MAT and PCR is necessary for a better diagnosis of the disease.

Serological evidence of exposure to Bartonella sp. in dogs with suspected vector-borne diseases, toxoplasmosis and neosporosis / Evidência sorológica de exposição a Bartonella sp. em cães com suspeita de doenças transmitidas por vetores, toxoplasmose e neosporose

Bartonellosis is a vector-borne zoonotic disease with worldwide distribution that infect a broad spectrum of mammalian species. Despite the recent studies carried out in Brazil, information regarding Bartonella in dogs are scarce. Therefore, we performed a retrospective study to investigate the exposure to Bartonella sp. in dogs by indirect immunofluorescence assay (IFA). Three hundred and thirty-five archived serum samples from dogs previously tested for vector-borne pathogens, Toxoplasma gondii, and Neospora caninum were screened for the presence of IgG antibodies to Bartonella sp. All dogs originated from the Metropolitan region of Ribeirão Preto, northeast of the State of São Paulo. Twenty-eight samples (8.3%) were positive for Bartonella sp. at the cut-off of 64. Among the 28 seropositive samples for Bartonella sp., 16 (57.1%) were also seropositive for Ehrlichia canis, 12 (42.8%) for Babesia vogeli, five (17.8%) for T. gondii and three (10.7%) for L. infantum and N. caninum. Our results demonstrated that dogs sampled were exposed to Bartonella sp. Since all the animals sampled in the present study were from private owners, our findings demonstrate that these people may also be exposed to Bartonella sp. Further studies designed to assess whether the infection by other arthropod-borne pathogens such as B. vogeli and E. canis are risk factors for Bartonella infection are needed.(AU)

A bartonelose é uma zoonose transmitida por vetores, com distribuição mundial, que infecta várias espécies de mamíferos. Apesar dos estudos conduzidos no Brasil, informações sobre Bartonella em cães são escassas. Portanto, foi realizado um estudo retrospectivo para investigar a exposição a Bartonella sp. em cães cães, utilizando-se o ensaio de imunofluorescência indireta (IFA). Trezentas e trinta e cinco amostras de soro de cães, previamente testadas para patógenos transmitidos pelos vetores, Toxoplasma gondii e Neospora caninum foram avaliadas quanto à presença de anticorpos IgG contra Bartonella sp. Todos os cães eram oriundos da região metropolitana de Ribeirão Preto, Nordeste do Estado de São Paulo. Vinte e oito amostras (8,3%) foram positivas para Bartonella sp. no ponto de corte de 64. Entre as 28 amostras positivas, 16 (57,1%) também foram positivas para Ehrlichia canis, 12 (42,8%) para Babesia vogeli, cinco (17,8%) para T. gondii e três (10,7%) para L. infantum e N. caninum. Os resultados demonstraram que os cães amostrados foram expostos a Bartonella sp. Como os animais eram de proprietários particulares, nossos achados demonstram que as pessoas também podem ter sido expostas a Bartonella sp. São necessários estudos para avaliar se a infecção por B. canis ou E. canis constitui fator de risco para infecção por Bartonella em cães.(AU)

Diagnóstico de leptospira spp. en bovinos sacrificados en brasil / Diagnostic of leptospira spp. in slaughtered cattle from brazil / Diagnóstico de leptospira spp. em bovinos abatidos no brasil

La leptospirosis es considerada una zoonosis bacteriana de importancia en salud pública. Es común en áreas tropicales, especialmente en países en desarrollo con escasos recursos de salud y saneamiento. Este estudio evaluó la presencia de Leptospira spp. en bovinos sacrificados en un matadero de la región Centro-Oeste de São Paulo, Brasil, así como animales positivos identificados tanto en serología como por análisis molecular. Se investigaron muestras biológicas de sangre, hígado y riñones de 150 bovinos mediante la técnica de Prueba de Aglutinación Microscópica (MAT) y Reacción en Cadena de la Polimerasa convencional (cPCR). Los resultados serológicos mostraron que de los 150 animales, 71 (47,3%) fueron reactivos. Los resultados moleculares mostraron la presencia de Leptospira spp. en riñones de 21 (14%) animales, en hígado de cinco (3,3%) animales, en hígado y riñones en dos animales (1,3%) y en sangre, en un (0,7%) animal. Estos resultados indican una alerta sobre la salud de los bovinos de carne debido a la posibilidad de que estos animales sean fuente de infección y la importancia de la característica ocupacional de esta enfermedad. También se verificó la importancia de complementar las técnicas serológicas y moleculares.  

Leptospirosis is considered a bacterial zoonosis of public health importance. It is common in tropical areas, especially in developing countries with scarce health and sanitation resources. This study evaluated the presence of Leptospira spp. in slaughtered bovine in a slaughterhouse in the Midwest region of São Paulo, Brazil, as well as identified positive animals both in serology and by molecular analysis. Biological samples of blood, liver and kidneys from 150 cattle were investigated by the technique of Microscopic Agglutination Test (MAT) and conventional Polymerase Chain Reaction (cPCR). The serological results showed that of the 150 animals, 71 (47.3%) were reactive. The molecular results showed the presence of Leptospira spp. in kidneys of 21 (14%) animals, in liver of five (3.3%) animals, in liver and kidneys in two animals (1.3%) and in blood, in one (0.7%) animal. These results indicate a warning about the health of beef cattle due to the possibility of these animals being the source of infection and the importance of the occupational characteristic of this disease. It was also verified the importance of complementing serological and molecular techniques.

A leptospirose é considerada uma zoonose bacteriana de importância para a saúde pública. É comum em áreas tropicais, especialmente em países em desenvolvimento com escassos recursos de saúde e saneamento. Este estudo avaliou a presença de Leptospira spp. em bovinos abatidos em frigorífico da região Centro-Oeste de São Paulo, Brasil, e identificou animais positivos tanto por sorologia quanto em análise molecular. Amostras biológicas de sangue, fígado e rins de 150 bovinos foram investigadas pela técnica de Teste de Aglutinação Microscópica (MAT) e Reação em Cadeia da Polimerase convencional (cPCR). Os resultados sorológicos mostraram que dos 150 animais, 71 (47,3%) foram reativos. Os resultados moleculares mostraram a presença de Leptospira spp. nos rins de 21 animais (14%), no fígado de 5 animais (3,3%), no fígado e rins em 2 animais (1,3%) e no sangue em 1 animal (0,7%). Esses resultados indicam um alerta sobre a saúde dos bovinos de corte devido à possibilidade desses animais serem fonte de infecção e a importância da característica ocupacional desta doença. Verificou-se também a importância de complementar as técnicas sorológicas e moleculares.

Seroprevalence and risk factors associated with seropositivity for equine encephalomyelitis virus in horses in Rio Grande do Norte, Brazil

The present objective was to investigate the presence of anti-equine viral encephalomyelitis (EVE) antibodies and the possible risk factors for its dissemination in horses raised in the East and West Potiguar mesoregions of the state of Rio Grande do Norte, Brazil. Serological diagnosis for neutralizing antibodies against Eastern (EEEV), Western (WEEV) and Venezuelan (VEEV). Equine viral encephalomyelitis was performed using a seroneutralization technique on 811 blood samples from horses from ninety properties and sixteen municipalities between July 2018 and February 2019. Factors associated with EVE were evaluated using an investigative epidemiological questionnaire, and the data were statistically analyzed using the Epi Info 3.5.2 software with a confidence level of 95%. The seroprevalence of anti-EVE antibodies was 14.2% (115), with 10.36% (84) for EEEV, 6.9% (56) for WEEV, and null for EVE. When analyzing risk factors, it can be concluded that horses raised in properties that do not clean installations and/or rent out their pasture are more likely to have anti-EVE antibodies. These results show evidence that horses raised in the East and West Potiguar mesoregions were exposed to EEEV and WEEV, thus reinforcing the importance of vaccination and serological survey of nonvaccinated horses as a means of monitoring the disease.

Canine visceral leishmaniasis in Rio Grande do Norte state, Northeastern Brazil - spatial analysis

Background: Human visceral leishmaniasis (HVL) is a potentially fatal disease with a worldwide distribution, being endemic in 12 countries in the Americas. The main reservoir in the urban environment is the dog, whose cases precede the disease in humans. For the control of HVL, the Ministry of Health of Brazil recommends efficiency in the notification of human cases, control of sandflies, elimination of reservoirs and health education, in addition to the interruption in the transmission of the disease by the intensification of surveillance and control of priority areas based on identification by spatial analysis. The objective of the study was to investigate the spatial distribution of canine visceral leishmaniasis (CVL) in the state of Rio Grande do Norte, Brazil, determining areas of risk by identifying spatial clusters, with a view to monitoring and implementing preventive actions. Materials, Methods & Results: Secondary data from sample and/or routine serological surveys for serological diagnosis of LVC in the period from 2011 to 2018 were used. The inclusion of animals in the routine diagnosis per municipality resulted from demands of veterinarians, veterinary clinics, dog tutors, zoonoses control centers and environmental surveillance. The spatial statistical analysis was performed with SatScan software version 9.6 for the detection of spatial clusters, based on using the statistical scan method. Of the total of 231,123 dogs tested in the period, 24,642 (10.6%) were seroreactive for CVL. During the study, the municipalities with the highest number of cases were Natal and Mossoró, with 9,671 and 4,514 cases, respectively. During the years 2011 to 2018, 38 significant clusters (P < 0.05) were identified that included one or more municipalities. Discussion: The state of Rio Grande do Norte has an urban environment susceptible to the occurrence of CVL, with climate and topography that favor the proliferation of the vector and housing in precarious socio-sanitary conditions. The high number of CVL cases in Natal can be explained by the fact that the city is considered endemic for CVL, characterized as an area of intense transmission of the disease, according to the Ministry of Health. In addition, public infrastructure in some locations is deficient and living conditions are unfavorable, so that there is a need to invest in effective protection measures for vector control, as well as a focus on health education, whose HVL control measures in the municipality need to be readjusted. The high rate of cases and the constant presence of clusters in the municipality of Açu can be explained by the increasing degradation of the Caatinga biome, evidenced by the removal of firewood for use in the ceramist pole, whose activity is concentrated on a large scale in the use of raw material and energy, through the production of charcoal, for agricultural and livestock fronts, putting species of fauna and flora at risk. It is also noteworthy that this fact contributes to the destruction of wild ecotopes, resulting in the search for the vector for other sources of human and animal food, allowing an increase in the number of cases of the disease. It's concluded that canine visceral leishmaniasis is distributed in a large part of the state of Rio Grande do Norte. The underreporting and/or deficiency in the disclosure of data by some municipalities represents a challenge in complying with the actions of the Visceral Leishmaniasis Surveillance and Control Program, and attention should be paid to the monitoring and inspection of the execution actions of municipal managers, as well as how to train professionals who are part of the service.

Blue tongue virus in dairy cattle in the Southern region of Minas Gerais, Brazil - serological survey

Background: Blue tongue (BT) is a noncontagious viral disease transmitted by hematophagous arthropods, especially of the genus Culicoides. The economic impact of the disease is related not only to deaths in sheep herds but also to the possible correlation of virus infection with the development of other diseases, such as pneumonia, abortion and movement problems. The economic losses caused by Blue Tongue are linked to restrictions on the import and export of animals and their genetic material and to the reproductive disorders associated with this disease. In addition, the fact that cattle take the role of reservoir, combined with the care by other countries with outbreaks of infection and biological contamination of their products, hinders trade in Mercosul, United States and Europe. Cattle are affected by Blue Tongue Virus in endemic areas and in some epidemic areas, but the development of clinical disease is rare. The clinical signs, when evident, range from reproductive losses, such as embryonic death, abortion, fetal malformation, temporary sterility, infertility in bulls, stillbirths and the birth of weak animals. The objective of this study was to determine the epidemiological aspects of Blue Tongue Virus (BTV) infection in dairy cattle in the Lavras region, state of Minas Gerais, Brazil. Materials, Methods & Results: A cross-sectional study was conducted to evaluate the frequency of cattle and herds seropositive for Blue Tongue in the southern region of Minas Gerais. In this study, 54 dairy farms were visited. A total of 586 serum samples were collected from cows of reproductive age. Sampling was random, and serum samples were collected from lactating cows over 24 months of age by puncture of the jugular vein and/or coccidian vein. The samples were transported and stored at the Setor de Patologia Veterinária, at the Universidade Federal de Lavras (SPV-UFLA), where they were centrifuged, and the serum aliquots were obtained, transferred to microtubes and kept at -20°C until the serological tests were performed. The samples were tested with the agarose gel immunodiffusion test (AGID) for anti-blue tongue virus antibodies. The AGID test is more practical and is the main method used to identify Blue Tongue Virus seroprevalence in different ruminant species. They are considered important tools for epidemiological surveillance of the disease. A prevalence of 83.28% was observed among animals that were seropositive for Blue Tongue Virus (488/586; IC 95% = 80.0 - 86.21). In addition, 100% (54/54; IC 95% = 93.4 - 100.0) of the farms had at least 1 positive animal, with rates ranging from 45.45% to 100% within the herds and where 22.22% of the farms had rates of 100% of the animals being positive. Discussion: Blue Tongue is a disease known to affect domestic and wild ruminants in Brazil. However, there is a lack of more precise information about its epidemiology and occurrence in the country and of joint efforts of researchers, producers and the government to understand in detail both the biology of vectors and the viral biology of Blue Tongue Virus in Brazil. This is the first record of detection of anti-blue tongue virus antibodies in cattle in the southern region of Minas Gerais. The results suggest that Blue Tongue Virus is present in cattle in the study area.

Multiparametric assay of screening for the diagnosis of mycoses of interest in Public Health: standardization of methodology / Ensaio multiparamétrico de triagem para diagnóstico de micoses de interesse em Saúde Pública: padronização da metodologia

The standardization and validation of a multiplex assay requires the combination of important parameters such as sensitivity and specificity, acceptable levels of performance, robustness, and reproducibility. We standardized a multiparametric Dot-blot aimed at the serological screening of paracoccidioidomycosis, histoplasmosis, and aspergillosis. A total of 148 serum were evaluated: 10 from healthy subjects, 36 from patients with paracoccidioidomycosis, 62 from patients with histoplasmosis, and 40 from patients with aspergillosis. It was found that the multiparametric Dot-blot showed a high percentage of cross-reactivity. However, when evaluated individually, in the serological screening of histoplasmosis, a good performance was observed when compared to the double immunodiffusion assay, considered the gold standard test, with 100% co-positivity and 83.3% co-negativity. The performance of serological screening for aspergillosis was not satisfactory when compared to double immunodiffusion, showing 71.4% co-positivity and 100% co-negativity. The evaluation of the stability of nitrocellulose membranes showed that membranes sensitized with H. capsulatum antigen remained stable for 90 days and those sensitized with A. fumigatus antigen for 30 days. We conclude that the use of crude antigens was not suitable for the standardization of the multiparametric Dot-blot assay, due to the high cross-reactivity, and that further tests should be performed with purified proteins (AU).

A padronização e validação de um ensaio multiplex requer a combinação de parâmetros importantes, como sensibilidade e especificidade, níveis aceitáveis de desempenho, robustez e reprodutibilidade. Este trabalho padronizou um Dot-blot multiparamétrico visando a triagem sorológica da paracoccidioidomicose, histoplasmose e aspergilose. Foram avaliadas 148 amostras de soro: 10 de indivíduos saudáveis, 36 de pacientes com paracoccidioidomicose, 62 de pacientes com histoplasmose e 40 de pacientes com aspergilose. Verificou-se que o Dot-blot multiparamétrico apresentou elevado percentual de reatividade cruzada. Entretanto, quando avaliado individualmente, na triagem sorológica da histoplasmose observou-se bom desempenho quando comparado ao ensaio de imunodifusão dupla, considerado o teste padrão ouro, com 100% de co-positividade e 83,3% de co-negatividade. O desempenho da triagem sorológica da aspergilose não foi satisfatório quando comparado a imunodifusão dupla, apresentando 71,4% de co-positividade e 100% de co-negatividade. A avaliação da estabilidade das membranas de nitrocelulose mostrou que membranas sensibilizadas com antígeno de H. capsulatum permaneceram estáveis por 90 dias e as sensibilizadas com antígeno de A. fumigatus, por 30 dias. Concluímos que o uso de antígenos brutos não foi adequado para a padronização do ensaio de Dot-blot multiparamétrico, devido ao alto índice de reatividade cruzada, e que novos testes devem ser realizados com proteínas purificadas (AU).