Bioinformatics and expression analysis of the Xeroderma Pigmentosum complementation group C (XPC) of Trypanosoma evansi in Trypanosoma cruzi cells / Análises de bioinformática e da expressão do gene Xeroderma Pigmentosum complementation group C (XPC) de Trypanosoma evansi em células de Trypanosoma cruzi

Abstract Nucleotide excision repair (NER) acts repairing damages in DNA, such as lesions caused by cisplatin. Xeroderma Pigmentosum complementation group C (XPC) protein is involved in recognition of global genome DNA damages during NER (GG-NER) and it has been studied in different organisms due to its importance in other cellular processes. In this work, we studied NER proteins in Trypanosoma cruzi and Trypanosoma evansi, parasites of humans and animals respectively. We performed three-dimensional models of XPC proteins from T. cruzi and T. evansi and observed few structural differences between these proteins. In our tests, insertion of XPC gene from T. evansi (TevXPC) in T. cruzi resulted in slower cell growth under normal conditions. After cisplatin treatment, T. cruzi overexpressing its own XPC gene (TcXPC) was able to recover cell division rates faster than T. cruzi expressing TevXPC gene. Based on these tests, it is suggested that TevXPC (being an exogenous protein in T. cruzi) interferes negatively in cellular processes where TcXPC (the endogenous protein) is involved. This probably occurred due interaction of TevXPC with some endogenous molecules or proteins from T.cruzi but incapacity of interaction with others. This reinforces the importance of correctly XPC functioning within the cell.

Resumo O reparo por excisão de nucleotídeos (NER) atua reparando danos no DNA, como lesões causadas por cisplatina. A proteína Xeroderma Pigmentosum complementation group C (XPC) está envolvida no reconhecimento de danos pela via de reparação global do genoma pelo NER (GG-NER) e tem sido estudada em diferentes organismos devido à sua importância em outros processos celulares. Neste trabalho, estudamos proteínas do NER em Trypanosoma cruzi e Trypanosoma evansi, parasitos de humanos e animais, respectivamente. Modelos tridimensionais das proteínas XPC de T. cruzi e T. evansi foram feitos e observou-se poucas diferenças estruturais entre estas proteínas. Durante testes, a inserção do gene XPC de T. evansi (TevXPC) em T. cruzi resultou em crescimento celular mais lento em condições normais. Após o tratamento com cisplatina, T. cruzi superexpressando seu próprio gene XPC (TcXPC) foi capaz de recuperar as taxas de divisão celular mais rapidamente do que T. cruzi expressando o gene TevXPC. Com base nesses testes, sugere-se que TevXPC (sendo uma proteína exógena em T. cruzi) interfere negativamente nos processos celulares em que TcXPC (a proteína endógena) está envolvida. Isso provavelmente ocorreu pois TevXPC é capaz de interagir com algumas moléculas ou proteínas endógenas de T.cruzi, mas é incapaz de interagir com outras. Isso reforça a importância do correto funcionamento de XPC dentro da célula.

Bioinformatics and expression analysis of the Xeroderma Pigmentosum complementation group C (XPC) of Trypanosoma evansi in Trypanosoma cruzi cells / Análises de bioinformática e da expressão do gene Xeroderma Pigmentosum complementation group C (XPC) de Trypanosoma evansi em células de Trypanosoma cruzi

Nucleotide excision repair (NER) acts repairing damages in DNA, such as lesions caused by cisplatin. Xeroderma Pigmentosum complementation group C (XPC) protein is involved in recognition of global genome DNA damages during NER (GG-NER) and it has been studied in different organisms due to its importance in other cellular processes. In this work, we studied NER proteins in Trypanosoma cruzi and Trypanosoma evansi, parasites of humans and animals respectively. We performed three-dimensional models of XPC proteins from T. cruzi and T. evansi and observed few structural differences between these proteins. In our tests, insertion of XPC gene from T. evansi (TevXPC) in T. cruzi resulted in slower cell growth under normal conditions. After cisplatin treatment, T. cruzi overexpressing its own XPC gene (TcXPC) was able to recover cell division rates faster than T. cruzi expressing TevXPC gene. Based on these tests, it is suggested that TevXPC (being an exogenous protein in T. cruzi) interferes negatively in cellular processes where TcXPC (the endogenous protein) is involved. This probably occurred due interaction of TevXPC with some endogenous molecules or proteins from T. cruzi but incapacity of interaction with others. This reinforces the importance of correctly XPC functioning within the cell.

O reparo por excisão de nucleotídeos (NER) atua reparando danos no DNA, como lesões causadas por cisplatina. A proteína Xeroderma Pigmentosum complementation group C (XPC) está envolvida no reconhecimento de danos pela via de reparação global do genoma pelo NER (GG-NER) e tem sido estudada em diferentes organismos devido à sua importância em outros processos celulares. Neste trabalho, estudamos proteínas do NER em Trypanosoma cruzi e Trypanosoma evansi, parasitos de humanos e animais, respectivamente. Modelos tridimensionais das proteínas XPC de T. cruzi e T. evansi foram feitos e observou-se poucas diferenças estruturais entre estas proteínas. Durante testes, a inserção do gene XPC de T. evansi (TevXPC) em T. cruzi resultou em crescimento celular mais lento em condições normais. Após o tratamento com cisplatina, T. cruzi superexpressando seu próprio gene XPC (TcXPC) foi capaz de recuperar as taxas de divisão celular mais rapidamente do que T. cruzi expressando o gene TevXPC. Com base nesses testes, sugere-se que TevXPC (sendo uma proteína exógena em T. cruzi) interfere negativamente nos processos celulares em que TcXPC (a proteína endógena) está envolvida. Isso provavelmente ocorreu pois TevXPC é capaz de interagir com algumas moléculas ou proteínas endógenas de T. cruzi, mas é incapaz de interagir com outras. Isso reforça a importância do correto funcionamento de XPC dentro da célula.

Bioinformatics and expression analysis of the Xeroderma Pigmentosum complementation group C (XPC) of Trypanosoma evansi in Trypanosoma cruzi cells / Análises de bioinformática e da expressão do gene Xeroderma Pigmentosum complementation group C (XPC) de Trypanosoma evansi em células de Trypanosoma cruzi

Nucleotide excision repair (NER) acts repairing damages in DNA, such as lesions caused by cisplatin. Xeroderma Pigmentosum complementation group C (XPC) protein is involved in recognition of global genome DNA damages during NER (GG-NER) and it has been studied in different organisms due to its importance in other cellular processes. In this work, we studied NER proteins in Trypanosoma cruzi and Trypanosoma evansi, parasites of humans and animals respectively. We performed three-dimensional models of XPC proteins from T. cruzi and T. evansi and observed few structural differences between these proteins. In our tests, insertion of XPC gene from T. evansi (TevXPC) in T. cruzi resulted in slower cell growth under normal conditions. After cisplatin treatment, T. cruzi overexpressing its own XPC gene (TcXPC) was able to recover cell division rates faster than T. cruzi expressing TevXPC gene. Based on these tests, it is suggested that TevXPC (being an exogenous protein in T. cruzi) interferes negatively in cellular processes where TcXPC (the endogenous protein) is involved. This probably occurred due interaction of TevXPC with some endogenous molecules or proteins from T. cruzi but incapacity of interaction with others. This reinforces the importance of correctly XPC functioning within the cell.(AU)

O reparo por excisão de nucleotídeos (NER) atua reparando danos no DNA, como lesões causadas por cisplatina. A proteína Xeroderma Pigmentosum complementation group C (XPC) está envolvida no reconhecimento de danos pela via de reparação global do genoma pelo NER (GG-NER) e tem sido estudada em diferentes organismos devido à sua importância em outros processos celulares. Neste trabalho, estudamos proteínas do NER em Trypanosoma cruzi e Trypanosoma evansi, parasitos de humanos e animais, respectivamente. Modelos tridimensionais das proteínas XPC de T. cruzi e T. evansi foram feitos e observou-se poucas diferenças estruturais entre estas proteínas. Durante testes, a inserção do gene XPC de T. evansi (TevXPC) em T. cruzi resultou em crescimento celular mais lento em condições normais. Após o tratamento com cisplatina, T. cruzi superexpressando seu próprio gene XPC (TcXPC) foi capaz de recuperar as taxas de divisão celular mais rapidamente do que T. cruzi expressando o gene TevXPC. Com base nesses testes, sugere-se que TevXPC (sendo uma proteína exógena em T. cruzi) interfere negativamente nos processos celulares em que TcXPC (a proteína endógena) está envolvida. Isso provavelmente ocorreu pois TevXPC é capaz de interagir com algumas moléculas ou proteínas endógenas de T. cruzi, mas é incapaz de interagir com outras. Isso reforça a importância do correto funcionamento de XPC dentro da célula.(AU)

Molecular detection and phylogenetic analysis of Trypanosoma spp. in Neotropical primates from Rio de Janeiro State, Brazil / Detecção molecular e análise filogenética de Trypanosoma spp. em primatas neotropicais do estado do Rio de Janeiro, Brasil

Trypanosoma spp. infection is a problem in many tropical countries, infecting several animal species, including humans. The aim of the present study was to identify the Trypanosoma species in Neotropical primates from Rio de Janeiro state and compare the results with other reports both phylogenetically and geographically. Molecular detection was based on the 18 SSU gene. The sequences obtained in the PCR were sequenced and compared with others previously deposited in GenBank. These sequences were used to perform phylogenetic analysis and make a distribution map of primate species infected by Trypanosoma species in Brazil. Among 34 monkeys, five capuchin monkeys (Sapajus spp.) and one marmoset (Callithrix spp.) showed Trypanosoma spp. sequences in the same clade of Trypanosoma minasense and three capuchin monkeys' sequences were in the same clade of Trypanosoma cruzi. The Atlantic Forest and the Brazilian Amazon are the regions with the highest frequency of studies about Trypanosoma spp. and variety of Neotropical primate hosts. These are areas that deserve attention regarding the conservation of biodiversity, but it also makes evident the lack of studies with Neotropical primates in other regions of the country, as well as multidisciplinary studies to better understand the host pathogen relationships.

A infecção por Trypanosoma spp. é um problema em muitos países tropicais, infectando várias espécies animais, incluindo humanos. O objetivo do presente estudo foi identificar as espécies de Trypanosoma em primatas neotropicais no estado Rio de Janeiro e comparar os resultados com outros relatos, tanto filogeneticamente quando geograficamente. A detecção molecular foi baseada no gene SSU 18. As sequências obtidas na PCR foram sequenciadas e comparadas com outras previamente depositadas no GenBank. Essas sequências foram utilizadas para análises filogenéticas e confeccionar um mapa de distribuição de espécies de primatas infectadas por espécies de Trypanosoma no Brasil. Entre 34 macacos, cinco macacos-prego (Sapajus spp.) e um sagui (Callithrix spp.) apresentaram sequências de Trypanosoma spp. no mesmo clado de Trypanosoma minasense e três sequências de macacos-prego estavam no mesmo clado de Trypanosoma cruzi. A Mata Atlântica e a Amazônia brasileira são as regiões com maior frequência de estudos sobre Trypanosoma spp. e variedade de primatas neotropicais hospedeiros. São áreas que merecem atenção no que se refere à conservação da biodiversidade, mas também evidencia a carência de estudos com PNH em outras regiões do país e de estudos multidisciplinares para melhor compreender as relações do patógeno hospedeiro.

Doença de chagas em cães / Chagas disease in dogs

A doença de Chagas, também chamada de tripanossomíase americana, é uma antropozoonose causada por um protozoário flagelado denominado Trypanosoma cruzi. A transmissão ocorre de diversas maneiras, sendo as mais relatadas: a forma vetorial, que ocorre quando o paciente inocula as fezes do vetor infectadas pelo tripanossomo por meio do orifício que foi aberto no momento do repasto sanguíneo, ou por via oral, devido principalmente a ingestão de alimentos contaminados com o tripanossomo. Diferentes espécies de animais domésticos e silvestres podem ser acometidas pela doença; contudo, o objetivo deste trabalho é descrever a doença em cães os quais são capazes de desenvolver o curso crônico, assim como os humanos, e são importantes marcadores de saúde pública para a doença, tendo em vista que são animais considerados sentinelas para o seu aparecimento.

Chagas disease, also called American trypanosomiasis, is an anthropozoonosis caused by the flagellated protozoan Trypanosoma cruzi. The transmission occurs in several ways, and the most reported are: the vector form, which occurs when the patient inoculates the feces of the vector infected by the trypanosome, through the hole that was opened during the blood meal, or orally, mainly due to the ingestion of food contaminated with trypanosome. Different species of domestic and wild animals can be affected by the disease; however, this study aims to describe the disease in dogs, which are capable of developing the chronic course, just like humans, and are also important public health indicators for the disease, since they are animals considered sentinels to its appearance.

Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation

Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect and the main vector of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). In the present study, the authors investigated whether a serine protease activity from the saliva of T. infestans has a role in vasomotor modulation, and in the insect-blood feeding by cleaving and activating protease-activated receptors (PARs). Methods T. infestans saliva was chromatographed as previously reported for purification of triapsin, a serine protease. The cleavage activity of triapsin on PAR peptides was investigated based on FRET technology. Mass spectrometry was used to analyze the sites of PAR-2 peptide cleaved by triapsin. NO measurements were performed using the DAN assay (2,3-diaminonapthalene). The vasorelaxant activity of triapsin was measured in vessels with or without functional endothelium pre-contracted with phenylephrine (3 µM). Intravital microscopy was used to assess the effect of triapsin on mouse skin microcirculation. Results Triapsin was able to induce hydrolysis of PAR peptides and showed a higher preference for cleavage of the PAR-2 peptide. Analysis by mass spectrometry confirmed a single cleavage site, which corresponds to the activation site of the PAR-2 receptor. Triapsin induced dose-dependent NO release in cultured human umbilical vein endothelial cells (HUVECs), reaching a maximum effect at 17.58 nM. Triapsin purified by gel-filtration chromatography (10-16 to 10-9 M) was applied cumulatively to mouse mesenteric artery rings and showed a potent endothelium-dependent vasodilator effect (EC30 = 10-12 M). Nitric oxide seems to be partially responsible for this vasodilator effect because L-NAME (L-NG-nitroarginine methyl ester 300 µM), a nitric oxide synthetase inhibitor, did not abrogate the vasodilation activated by triapsin. Anti-PAR-2 antibody completely inhibited vasodilation observed in the presence of triapsin activity. Triapsin activity also induced an increase in the mouse ear venular diameter. Conclusion Data from this study suggest a plausible association between triapsin activity mediated PAR-2 activation and vasodilation caused by T. infestans saliva.(AU)

Validation of a documentary on Chagas disease by a population living in an endemic area / Validação de um documentário sobre a doença de Chagas por uma população de área endêmica

Educational interventions may trigger actions that contribute to prevent parasitic diseases, such as Chagas disease (CD). This study aimed at investigating the impact of an instructional video named Documentary on Chagas Disease on knowledge about CD and its vectors displayed by a population that lives in an endemic area in Brazil, so as to validate it as an educational tool. The video was shown to 226 subjects, divided into two groups. Group 1 was composed of users of Basic Health Units (BHU) in Pelotas and Pinheiro Machado, cities located in Rio Grande do Sul (RS) state, Brazil, where CD is endemic. Group 2 consisted of students who attend three public schools located in the rural area in Pinheiro Machado, RS. Two questionnaires with questions about their knowledge about triatomines and CD were applied, before and after the documentary was shown. After the video was shown, there was significant increase in individuals knowledge (in both groups) about kissing bugs, their notification, the disease and its prevention. Besides, watchers considered that the quality of the material was satisfactory. Since the Documentary on CD can be easily accessed on the internet and was effective in teaching the population that lives in endemic areas, its use should be encouraged in places and meetings connected to health that aim at fighting against triatominae and at exposing an updated view of CD.(AU)

Intervenções educacionais podem desencadear ações que contribuam para a prevenção de doenças parasitárias, como a doença de Chagas (DC). Este estudo teve como objetivo investigar o impacto de um vídeo instrucional denominado Documentário Doença de Chagas no conhecimento sobre DC e seus vetores exibidos a uma população que vive em área endêmica no Brasil, para validá-lo como uma ferramenta educacional. O vídeo foi exibido para 226 indivíduos, divididos em dois grupos. O grupo 1 foi composto por usuários das Unidades Básicas de Saúde (UBS) de Pelotas e Pinheiro Machado, cidades do estado do Rio Grande do Sul (RS), Brasil, onde a DC é endêmica. O grupo 2 foi formado por estudantes que frequentam três escolas públicas localizadas na zona rural de Pinheiro Machado, RS. Dois questionários com perguntas sobre conhecimentos sobre triatomíneos e DC foram aplicados, antes e depois da exibição do documentário. Após a exibição do vídeo, houve um aumento significativo no conhecimento dos indivíduos (em ambos os grupos) sobre triatomíneos, sua notificação, a doença e sua prevenção. Além disso, os observadores consideraram que a qualidade do material era satisfatória. Como o Documentário DC pode ser facilmente acessado na Internet e ser eficaz no ensino à população que vive em áreas endêmicas, seu uso deve ser incentivado em locais e reuniões relacionadas à saúde, que visam combater os triatomíneos e exibir uma informação atualizada sobre a DC.(AU)

Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation

Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect and the main vector of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). In the present study, the authors investigated whether a serine protease activity from the saliva of T. infestans has a role in vasomotor modulation, and in the insect-blood feeding by cleaving and activating protease-activated receptors (PARs). Methods T. infestans saliva was chromatographed as previously reported for purification of triapsin, a serine protease. The cleavage activity of triapsin on PAR peptides was investigated based on FRET technology. Mass spectrometry was used to analyze the sites of PAR-2 peptide cleaved by triapsin. NO measurements were performed using the DAN assay (2,3-diaminonapthalene). The vasorelaxant activity of triapsin was measured in vessels with or without functional endothelium pre-contracted with phenylephrine (3 µM). Intravital microscopy was used to assess the effect of triapsin on mouse skin microcirculation. Results Triapsin was able to induce hydrolysis of PAR peptides and showed a higher preference for cleavage of the PAR-2 peptide. Analysis by mass spectrometry confirmed a single cleavage site, which corresponds to the activation site of the PAR-2 receptor. Triapsin induced dose-dependent NO release in cultured human umbilical vein endothelial cells (HUVECs), reaching a maximum effect at 17.58 nM. Triapsin purified by gel-filtration chromatography (10-16 to 10-9 M) was applied cumulatively to mouse mesenteric artery rings and showed a potent endothelium-dependent vasodilator effect (EC30 = 10-12 M). Nitric oxide seems to be partially responsible for this vasodilator effect because L-NAME (L-NG-nitroarginine methyl ester 300 µM), a nitric oxide synthetase inhibitor, did not abrogate the vasodilation activated by triapsin. Anti-PAR-2 antibody completely inhibited vasodilation observed in the presence of triapsin activity. Triapsin activity also induced an increase in the mouse ear venular diameter. Conclusion Data from this study suggest a plausible association between triapsin activity mediated PAR-2 activation and vasodilation caused by T. infestans saliva.(AU)

Protocol optimization for the detection of Trypanosoma cruzi DNA in açai (Euterpe oleraceae) pulp / Otimização de protocolo para detecção de DNA de Trypanosoma cruzi em polpa de açaí (Euterpe oleracea)

Chagas disease, caused by the protozoan Trypanosoma cruzi, has often been linked to oral transmission through açai consumption. Molecular methods that allow fast and accurate identification of the pathogen are important for the detection of the presence of the parasite in this food. This study aimed to optimize polymerase chain reaction (PCR)-based detection of T. cruzi DNA in açai pulp. Several dilutions of T. cruzi DTU TcI trypomastigote forms were cultured in liver infusion tryptose (LIT) medium. Trypanosoma cruzi DNA was extracted from the cells and subjected to PCR. Subsequently, culture dilutions were added to açai pulp to evaluate the detection threshold of the optimized PCR assay. We demonstrate that our assay can detect T. cruzi DNA in açai pulp at a concentration of 1.08 × 10-10 ng µL-1. We conclude that our optimized protocol is effective and can be used as an important tool for the detection of T. cruzi contamination in açaí.(AU)

A doença de Chagas, enfermidade causada pelo protozoário Trypanosoma cruzi, tem sido relacionada com frequência à transmissão oral pelo consumo de açaí. Métodos moleculares que fornecem uma identificação rápida e precisa do patógeno para a detecção da presença do parasita são de extrema importância para a detecção da presença do parasita neste alimento. Este estudo teve como objetivo otimizar a detecção de DNA de T. cruzi em polpa de açaí por meio da reação em cadeia da polimerase (PCR). Foram preparadas várias diluições das formas tripomastigotas de T. cruzi DTU TcI cultivadas em meio de cultivo Liver Infusion Tryptose. O DNA de T. cruzi foi extraído das células e submetido à PCR. Posteriormente, as diluições da cultura foram adicionadas às polpas de açaí para avaliar o limite de detecção do novo ensaio de PCR otimizado. Mostramos que nosso ensaio pode detectar DNA de T. cruzi em polpas de açaí na concentração de 1.08 × 10-10 ng µL-1. Concluímos que a metodologia desenvolvida se mostra eficaz e pode ser uma ferramenta importante para a detecção de contaminação por T. cruzi em açaí.(AU)

Ocorrência de triatomíneos no Estado do Ceará, Brasil / Occurrence of triatomines in the state of Ceará, Brazil

Esta revisão integrativa tem como objetivo analisar as publicações científicas nos últimos doze anos sobre triatomíneos encontrados no Estado do Ceará. Para tal, nos bancos de dados da Biblioteca Virtual em Saúde (BVS) foi realizada uma busca com associação dos descritores “Doença de Chagas”, “Triatomíneos” e “Brasil”. Após aplicação dos critérios de inclusão, exclusão e da análise dos seus conteúdos, oito artigos realizados no Estado do Ceará foram selecionados para a análise do estudo. Algumas espécies de triatomíneos encontradas no Nordeste são de extrema importância epidemiológica para o homem, pois estão associadas com domicílios humanos. As espécies mais predominantes com maior importância epidemiológica, com distribuição ampla no Ceará e presente no ambiente peridoméstico foram: Triatoma brasiliensis, Triatoma pseudomaculata, Panstrogylus megistus, Panstrogylus lutzi e Rhodnius nasutus. Várias espécies de triatomíneos tem a capacidade de transmitir Trypanosoma cruzi e de se adaptar com o ambiente peridoméstico e doméstico tornando o homem mais suscetível a infecção, sendo assim necessário estratégias de vigilância e controle do vetor.

This integrative review aims to analyze the scientific publications in the last twelve years on triatomines found at Ceará State. For this, a search was performed in the Biblioteca Virtual em Saúde (BVS) databases with association of the descriptors “Chagas Disease”, “Triatomines” and “Brazil”. After applying the inclusion, exclusion criteria and analysis of their content, eight articles were selected for the analysis and discussion. Some species of triatomines found in the Northeast region are of extreme epidemiological importance since they are associated with human households. The most prevalent species with epidemiological importance due to their large distribution in Ceará and presence in the peridomestic environment are: Triatoma brasiliensis, Triatoma pseudomaculata, Panstrogylus megistus, Panstrogylus lutzi and Rhodnius nasutus. Several species of triatomines have the capacity to transmit T. cruzi and the ability to adapt to the peridomestic and domestic environment turning humans more susceptible to infection, thus requiring surveillance and vector control strategies.

Natural infection by Trypanosoma cruzi in a dog from Bahia state - northeast of Brazil

Background: Chagas Disease (CD) is a parasitic anthropozoonosis caused by the Trypanosoma cruzi (T. cruzi), a protozoan transmitted by insects from the Reduviidae family. Several species of wild and domestic animals, humans included, are susceptible, developing acute clinical signals (myocarditis and cardiac arrhythmias) or chronic signals (drop on the performance and syncope) of the disease. In Brazil, the disease in dogs shows variable indexes of occurrence. The present paper describes the natural infection by T. cruzi in a canine living in the city of Barra, in the state of Bahia, Brazil, a region classified as high risk of vulnerability for the CD in the northeast of Brazil. Case: A 9-year-old male dog, non-specific breed, was referred to the clinic the Veterinary Clinic for Small Animals, at the Veterinary Medical Teaching Hospital), at the Federal University of the West of Bahia. Its owner reported the presence of bloody secretion in the penile region. During the physical exam it was possible to observe an increase in the volume of the penis base. All of the animal’s physiological parameters were within normal levels. Blood samples and cytological laminas from the lesioned area were collected and sent to the Laboratory of Veterinary Clinical Pathology in the same institution. The cytological exam showed monomorphic population of great round cells with a round nucleus, condensed chromatin and one to two prominent nucleolus, abundant and slightly basophilic cytoplasm, with multiple stippled vacuoles, thus confirming a Transmissible venereal tumor (TVT) case. With the blood exam one observed a thrombocytopenia, neutrophilia with a deviation on the left and the presence of a trypomastigote form of Trypanosoma sp. in the blood smears...

Natural infection by Trypanosoma cruzi in a dog from Bahia state - northeast of Brazil

Background: Chagas Disease (CD) is a parasitic anthropozoonosis caused by the Trypanosoma cruzi (T. cruzi), a protozoan transmitted by insects from the Reduviidae family. Several species of wild and domestic animals, humans included, are susceptible, developing acute clinical signals (myocarditis and cardiac arrhythmias) or chronic signals (drop on the performance and syncope) of the disease. In Brazil, the disease in dogs shows variable indexes of occurrence. The present paper describes the natural infection by T. cruzi in a canine living in the city of Barra, in the state of Bahia, Brazil, a region classified as high risk of vulnerability for the CD in the northeast of Brazil. Case: A 9-year-old male dog, non-specific breed, was referred to the clinic the Veterinary Clinic for Small Animals, at the Veterinary Medical Teaching Hospital), at the Federal University of the West of Bahia. Its owner reported the presence of bloody secretion in the penile region. During the physical exam it was possible to observe an increase in the volume of the penis base. All of the animals physiological parameters were within normal levels. Blood samples and cytological laminas from the lesioned area were collected and sent to the Laboratory of Veterinary Clinical Pathology in the same institution. The cytological exam showed monomorphic population of great round cells with a round nucleus, condensed chromatin and one to two prominent nucleolus, abundant and slightly basophilic cytoplasm, with multiple stippled vacuoles, thus confirming a Transmissible venereal tumor (TVT) case. With the blood exam one observed a thrombocytopenia, neutrophilia with a deviation on the left and the presence of a trypomastigote form of Trypanosoma sp. in the blood smears...(AU)

Ocorrência de triatomíneos no Estado do Ceará, Brasil / Occurrence of triatomines in the state of Ceará, Brazil

Esta revisão integrativa tem como objetivo analisar as publicações científicas nos últimos doze anos sobre triatomíneos encontrados no Estado do Ceará. Para tal, nos bancos de dados da Biblioteca Virtual em Saúde (BVS) foi realizada uma busca com associação dos descritores “Doença de Chagas”, “Triatomíneos” e “Brasil”. Após aplicação dos critérios de inclusão, exclusão e da análise dos seus conteúdos, oito artigos realizados no Estado do Ceará foram selecionados para a análise do estudo. Algumas espécies de triatomíneos encontradas no Nordeste são de extrema importância epidemiológica para o homem, pois estão associadas com domicílios humanos. As espécies mais predominantes com maior importância epidemiológica, com distribuição ampla no Ceará e presente no ambiente peridoméstico foram: Triatoma brasiliensis, Triatoma pseudomaculata, Panstrogylus megistus, Panstrogylus lutzi e Rhodnius nasutus. Várias espécies de triatomíneos tem a capacidade de transmitir Trypanosoma cruzi e de se adaptar com o ambiente peridoméstico e doméstico tornando o homem mais suscetível a infecção, sendo assim necessário estratégias de vigilância e controle do vetor.(AU)

This integrative review aims to analyze the scientific publications in the last twelve years on triatomines found at Ceará State. For this, a search was performed in the Biblioteca Virtual em Saúde (BVS) databases with association of the descriptors “Chagas Disease”, “Triatomines” and “Brazil”. After applying the inclusion, exclusion criteria and analysis of their content, eight articles were selected for the analysis and discussion. Some species of triatomines found in the Northeast region are of extreme epidemiological importance since they are associated with human households. The most prevalent species with epidemiological importance due to their large distribution in Ceará and presence in the peridomestic environment are: Triatoma brasiliensis, Triatoma pseudomaculata, Panstrogylus megistus, Panstrogylus lutzi and Rhodnius nasutus. Several species of triatomines have the capacity to transmit T. cruzi and the ability to adapt to the peridomestic and domestic environment turning humans more susceptible to infection, thus requiring surveillance and vector control strategies.(AU)

Valores hematológicos de lobinhos (Cerdocyon thous) do Pantanal, Mato Grosso do Sul, Brasil naturalmente infectados e não infectados por Trypanosoma cruzi e T. evansi / Hematological values of crab eating foxes (Cerdocyon thous) from Pantanal, Mato Grosso do Sul, Brazil naturally infected and non-infected by Trypanosoma cruzi and T. evansi

O objetivo do trabalho foi avaliar a influência do sexo, da sazonalidade e de infecções por Trypanosoma cruzi e Trypanosoma evansi nos valores hematológicos de lobinhos (Cerdocyon thous) que habitam o Pantanal Sul-Mato-Grossense. Entre novembro de 2015 e outubro de 2016, foram amostrados 48 lobinhos. Os valores hematológicos mensurados foram as contagens de eritrócitos e leucócitos, volume globular e volume corpuscular médio. Consideramos como parasitados os animais positivos para T.cruzi e T. evansi em qualquer um dos testes diagnósticos utilizados. Observamos que sete (14.5%) lobinhos encontraram-se parasitados somente por T.cruzi, sete por T. evansi, e nove (19%) estavam coinfectados. Os animais parasitados por T.cruzi mostraram um aumento significativo das contagens de leucócitos (14.7 x 103) em relação aos animais não parasitados (10.4 x 103), parasitados por T. evansi (12.4 x 103) e coinfectados (12.9 x 103). Observamos diferença significativa em relação aos valores médios de eritrócitos e volume corpuscular médio entre os períodos de cheia e seca: 2.6 x 106 e 165, e 3.4 x 106 e 132, respectivamente. Todos apresentaram macrocitose não confirmada morfologicamente.

The aim of this work was to evaluate the influence of sex, seasonality and infections by Trypanosoma cruzi and Trypanosoma evansi on the hematological values of crab-eating foxes (Cerdocyon thous) that inhabit the Pantanal from Mato Grosso do Sul. Between November of 2015 and October of 2016, 48 animals were sampled. The hematological values measured were erythrocyte and leukocyte counts, globular volume and mean corpuscular volume. We considered as parasitized the positive animals for T.cruzi and T. evansi in any of diagnostic tests used. We observed that seven (14.5%) of the crab-eating foxes were parasitized only by T.cruzi, seven by T. evansi, and nine (19%) were coinfected. The parasitized animals by T.cruzi showed a significant increase in leukocyte counts (14.7 x 103) compared to non-parasitized animals (10.4 x 103), parasitized by T. evansi (12.4 x 103) and coinfected (12.9 x 103). We observed a significant difference in relation to mean erythrocyte and mean corpuscular volume values between wet and dry periods: 2.6 x 106 and 165, and 3.4 x 106 and 132, respectively. All presented morphologically unconfirmed macrocytosis.

Valores hematológicos de lobinhos (Cerdocyon thous) do Pantanal, Mato Grosso do Sul, Brasil naturalmente infectados e não infectados por Trypanosoma cruzi e T. evansi / Hematological values of crab eating foxes (Cerdocyon thous) from Pantanal, Mato Grosso do Sul, Brazil naturally infected and non-infected by Trypanosoma cruzi and T. evansi

O objetivo do trabalho foi avaliar a influência do sexo, da sazonalidade e de infecções por Trypanosoma cruzi e Trypanosoma evansi nos valores hematológicos de lobinhos (Cerdocyon thous) que habitam o Pantanal Sul-Mato-Grossense. Entre novembro de 2015 e outubro de 2016, foram amostrados 48 lobinhos. Os valores hematológicos mensurados foram as contagens de eritrócitos e leucócitos, volume globular e volume corpuscular médio. Consideramos como parasitados os animais positivos para T.cruzi e T. evansi em qualquer um dos testes diagnósticos utilizados. Observamos que sete (14.5%) lobinhos encontraram-se parasitados somente por T.cruzi, sete por T. evansi, e nove (19%) estavam coinfectados. Os animais parasitados por T.cruzi mostraram um aumento significativo das contagens de leucócitos (14.7 x 103) em relação aos animais não parasitados (10.4 x 103), parasitados por T. evansi (12.4 x 103) e coinfectados (12.9 x 103). Observamos diferença significativa em relação aos valores médios de eritrócitos e volume corpuscular médio entre os períodos de cheia e seca: 2.6 x 106 e 165, e 3.4 x 106 e 132, respectivamente. Todos apresentaram macrocitose não confirmada morfologicamente.(AU)

The aim of this work was to evaluate the influence of sex, seasonality and infections by Trypanosoma cruzi and Trypanosoma evansi on the hematological values of crab-eating foxes (Cerdocyon thous) that inhabit the Pantanal from Mato Grosso do Sul. Between November of 2015 and October of 2016, 48 animals were sampled. The hematological values measured were erythrocyte and leukocyte counts, globular volume and mean corpuscular volume. We considered as parasitized the positive animals for T.cruzi and T. evansi in any of diagnostic tests used. We observed that seven (14.5%) of the crab-eating foxes were parasitized only by T.cruzi, seven by T. evansi, and nine (19%) were coinfected. The parasitized animals by T.cruzi showed a significant increase in leukocyte counts (14.7 x 103) compared to non-parasitized animals (10.4 x 103), parasitized by T. evansi (12.4 x 103) and coinfected (12.9 x 103). We observed a significant difference in relation to mean erythrocyte and mean corpuscular volume values between wet and dry periods: 2.6 x 106 and 165, and 3.4 x 106 and 132, respectively. All presented morphologically unconfirmed macrocytosis.(AU)

Infecção por Toxoplasma gondii, Neospora caninum, Leishmania major e Trypanosoma cruzi em cães do estado do Pará / Infection by Toxoplasma gondii, Neospora caninum, Leishmania major and Trypanosoma cruzi in dogs from the state of Pará

Realizou-se inquérito sorológico para Toxoplasma gondii, Neospora caninum, Leishmania spp. e Trypanosoma cruzi em cães dos municípios de Castanhal e Belém, no Estado do Pará, e estudou-se a associação entre os animais sororeagentes e fatores como sexo, idade, acesso a rua e procedência dos animais. Foram utilizados 269 cães de diferentes idades, sadios, provenientes de abrigos de animais, canil comercial, Centro de Controle de Zoonoses e domiciliados. Utilizou-se a reação de imunofluorescência indireta (RIFI), com lâminas sensibilizadas de acordo com os seus respectivos antígenos. Das 263 amostras, 100 foram reagentes, com 38,0% de positividade para Toxoplasma gondii, sendo os títulos prevalentes 16UI e 64UI, 15,97% de sororreagentes para anticorpos anti-N. caninum com maior soroprevalência para os títulos 25UI e 50UI. Na pesquisa de anticorpos anti-L. major, 3,42% foram sororreagentes com o título 40UI, sendo o mais prevalente. A pesquisa de anticorpos anti-T. cruzi revelou negatividade em todas as amostras. As variáveis procedência e faixa etária mostraram ser um fator de risco para os animais. Os animais de Belém apresentaram 2,6 vezes mais chances de infecção do que os animais de Castanhal. Os animais na faixa etária de três a sete anos apresentaram 2,27 vezes mais chances de serem sororreagentes do que os cães com até três anos de idade. Não houve associação estatística para as demais variáveis e protozoários pesquisados.

A serological inquiry was performed to Toxoplasma gondii, Neospora caninum, Leishmania spp. and Trypanosoma cruzi in dogs from the city of Castanhal and Belém, State of Pará, and the association between seropositive animals and factors such as sex, age, access to the streets and provenance of the animals was studied. 269 healthy dogs with different ages, coming from animal shelters, commercial kennels, Zoonosis Control Centre, and domiciles were used. The Indirect immunofluorescence (IIF) reaction was used with plates sensitized with their respective antigens. Among the 263 samples, 100 were reactant, with 38.0% of positivity for Toxplasma gondii, being 16UI and 64UI the prevalent titers, 15.97% of reagent animals for anti-N. caninum antibodies with higher seroprevalence to titers 25UI and 50UI.At the anti-L. major antibodies research, 3.42% were reagent with the titer 40UI being the prevalent. The research of anti-T.cruzi antibodies revealed negativity in all the samples. The provenance and age range variables have been shown as a risk factor to the animals. The animals from the city of Belém presented 2.6 times more changes of infection than the animals from the city of Castanhal. The animals at the age range from three to seven years old presented 2.27 times more changes of being reagent than the dogs with up to three years old. There was no statistical association to the other variables and protozoa researched.

Infecção por Toxoplasma gondii, Neospora caninum, Leishmania major e Trypanosoma cruzi em cães do estado do Pará / Infection by Toxoplasma gondii, Neospora caninum, Leishmania major and Trypanosoma cruzi in dogs from the state of Pará

Realizou-se inquérito sorológico para Toxoplasma gondii, Neospora caninum, Leishmania spp. e Trypanosoma cruzi em cães dos municípios de Castanhal e Belém, no Estado do Pará, e estudou-se a associação entre os animais sororeagentes e fatores como sexo, idade, acesso a rua e procedência dos animais. Foram utilizados 269 cães de diferentes idades, sadios, provenientes de abrigos de animais, canil comercial, Centro de Controle de Zoonoses e domiciliados. Utilizou-se a reação de imunofluorescência indireta (RIFI), com lâminas sensibilizadas de acordo com os seus respectivos antígenos. Das 263 amostras, 100 foram reagentes, com 38,0% de positividade para Toxoplasma gondii, sendo os títulos prevalentes 16UI e 64UI, 15,97% de sororreagentes para anticorpos anti-N. caninum com maior soroprevalência para os títulos 25UI e 50UI. Na pesquisa de anticorpos anti-L. major, 3,42% foram sororreagentes com o título 40UI, sendo o mais prevalente. A pesquisa de anticorpos anti-T. cruzi revelou negatividade em todas as amostras. As variáveis procedência e faixa etária mostraram ser um fator de risco para os animais. Os animais de Belém apresentaram 2,6 vezes mais chances de infecção do que os animais de Castanhal. Os animais na faixa etária de três a sete anos apresentaram 2,27 vezes mais chances de serem sororreagentes do que os cães com até três anos de idade. Não houve associação estatística para as demais variáveis e protozoários pesquisados.(AU)

A serological inquiry was performed to Toxoplasma gondii, Neospora caninum, Leishmania spp. and Trypanosoma cruzi in dogs from the city of Castanhal and Belém, State of Pará, and the association between seropositive animals and factors such as sex, age, access to the streets and provenance of the animals was studied. 269 healthy dogs with different ages, coming from animal shelters, commercial kennels, Zoonosis Control Centre, and domiciles were used. The Indirect immunofluorescence (IIF) reaction was used with plates sensitized with their respective antigens. Among the 263 samples, 100 were reactant, with 38.0% of positivity for Toxplasma gondii, being 16UI and 64UI the prevalent titers, 15.97% of reagent animals for anti-N. caninum antibodies with higher seroprevalence to titers 25UI and 50UI.At the anti-L. major antibodies research, 3.42% were reagent with the titer 40UI being the prevalent. The research of anti-T.cruzi antibodies revealed negativity in all the samples. The provenance and age range variables have been shown as a risk factor to the animals. The animals from the city of Belém presented 2.6 times more changes of infection than the animals from the city of Castanhal. The animals at the age range from three to seven years old presented 2.27 times more changes of being reagent than the dogs with up to three years old. There was no statistical association to the other variables and protozoa researched.(AU)

Morphobiological, morphometric and ultrastructural characterization of sylvatic Trypanosoma cruzi isolates from Rio de Janeiro state, Brazil / Caracterização morfobiológica, morfométrica e ultraestrutural de isolados silvestres de Trypanosoma cruzi do estado do Rio de Janeiro, Brasil

Triatoma vitticeps is a triatomine with geographic distribution restrict to Brazil, which exhibits high prevalence of Trypanosoma cruzi natural infection. Of special epidemiologic concern, this species often invades households in the states of Rio de Janeiro, Minas Gerais and Espírito Santo. The objective of this study was to evaluate morphological and ultrastructural parameters on three T. cruzi isolates obtained from wild T. vitticeps specimens. The growth and cell differentiation of the parasite was evaluated through epimastigote and trypomastigote forms obtained in the growth curves for three distinct isolates. The maximum growth showed differences at the 20th day of the curve. Our in vitro results show a heterogeneity, regarding these features for samples cultivated under the same conditions. Morphometric analyzes based on the shape of epimastigotes and trypomastigotes corroborated such differentiation. These results highlight the need of better understanding the meaning of this diversity under an eco-epidemiological perspective.(AU)

Triatoma vitticeps é um triatomíneo com distribuição geográfica restrita ao território brasileiro, apresentando alta prevalência de infecção natural pelo Trypanosoma cruzi. Esta espécie é relevante sob o ponto de vista epidemiológico por invadir domicílios com frequência nos estados do Rio de Janeiro, Minas Gerais e Espírito Santo. O objetivo deste estudo foi avaliar parâmetros morfológicos e ultraestruturais, em três isolados de T. cruzi obtidos a partir de triatomíneos silvestres. O crescimento e a diferenciação celular do parasita foi avaliado através das formas epimastigotas e tripomastigotas obtidas nas curvas de crescimento para os três isolados. O crescimento máximo mostrou diferenças no 20º dia da curva. Nossos resultados in vitro mostram uma heterogeneidade, em relação a essas características para amostras cultivadas nas mesmas condições. As análises morfométricas baseadas na conformação de epimastigotas e trypomastigotes corroboraram essa diferenciação. Estes resultados ressaltam a necessidade de uma melhor compreensão do significado desta diversidade sob uma perspectiva eco-epidemiológica.(AU)

Using prodigiosin against some gram-positive and gram-negative bacteria and Trypanosoma cruzi

This work aimed to explore the action of natural prodigiosin on both bacterial organisms and Trypanosoma cruzi cells. Methods: Natural prodigiosin pigment was extracted and purified from cultures of Serratia marcescens. Two media, peanut broth and peptone glycerol broth, both recommended in the literature for prodigiosin production, were compared. The prodigiosin obtained was employed to explore its antimicrobial properties against both bacteria and Trypanosoma cruzi cells. Results: Peanut broth yielded four times more prodigiosin. The prodigiosin showed remarkable activity (minimal inhibitory concentrations in the range of 2-8 µM for bacteria and half maximal inhibitory concentration of 0.6 µM for Trypanosoma cruzi). In fact, the prodigiosin concentration required to inhibit parasite growth was as low as 0.25 mg/l versus 4.9 mg/l of benznidazole required. Furthermore, atomic force microscopy revealed marked morphological alterations in treated epimastigote forms, although no pore-formation activity was detected in protein-free environments. Conclusions: This work demonstrates the potential usefulness of prodigiosin against some gram-positive and gram-negative bacteria and Trypanosoma cruzi although further studies must be done in order to assess its value as a candidate molecule.(AU)

Efficacy of sertraline against Trypanosoma cruzi: an in vitro and in silico study

Drug repurposing has been an interesting and cost-effective approach, especially for neglected diseases, such as Chagas disease. Methods: In this work, we studied the activity of the antidepressant drug sertraline against Trypanosoma cruzi trypomastigotes and intracellular amastigotes of the Y and Tulahuen strains, and investigated its action mode using cell biology and in silico approaches. Results: Sertraline demonstrated in vitro efficacy against intracellular amastigotes of both T. cruzi strains inside different host cells, including cardiomyocytes, with IC50 values between 1 to 10 µM, and activity against bloodstream trypomastigotes, with IC50 of 14 µM. Considering the mammalian cytotoxicity, the drug resulted in a selectivity index of 17.8. Sertraline induced a change in the mitochondrial integrity of T. cruzi, resulting in a decrease in ATP levels, but not affecting reactive oxygen levels or plasma membrane permeability. In silico approaches using chemogenomic target fishing, homology modeling and molecular docking suggested the enzyme isocitrate dehydrogenase 2 of T. cruzi (TcIDH2) as a potential target for sertraline. Conclusions: The present study demonstrated that sertraline had a lethal effect on different forms and strains of T. cruzi, by affecting the bioenergetic metabolism of the parasite. These findings provide a starting point for future experimental assays and may contribute to the development of new compounds.(AU)