Fragment detection of Coleopteran and Triatomine insects in experimentally contaminated acai pulp and sugarcane juice.

INTRODUCTION: Oral transmission of acute Chagas disease is an emerging public health concern. This study aimed to detect insect fragments in experimentally contaminated food, by comparing triatomines with other insects. METHODS: Food samples were experimentally contaminated with insects, processed to recover their fragments by light filth, and analyzed by microscopy and Polymerase Chain Reaction (PCR). RESULTS: Morphological differences between coleopteran and triatomine insects were observed in microscopic images. PCR was efficient in amplifying Triatominae DNA in the experimentally contaminated food. CONCLUSIONS: This methodology could be utilized by food analysts to identify possible insect contamination in food samples.

Fragment detection of coleopteran and triatomine insects in experimentally contaminated acai pulp and sugarcane juice

INTRODUCTION: Oral transmission of acute Chagas disease is an emerging public health concern. This study aimed to detect insect fragments in experimentally contaminated food, by comparing triatomines with other insects. METHODS: Food samples were experimentally contaminated with insects, processed to recover their fragments by light filth, and analyzed by microscopy and Polymerase Chain Reaction (PCR). RESULTS: Morphological differences between coleopteran and triatomine insects were observed in microscopic images. PCR was efficient in amplifying Triatominae DNA in the experimentally contaminated food. CONCLUSIONS: This methodology could be utilized by food analysts to identify possible insect contamination in food samples. (AU)

Avaliação prognóstica do intervalo Tpico ­ T fim e intervalo QT corrigidos, em pacientes com Doença de Chagas, fase crônica e eletrocardiograma normal / Prognostic evaluation of corrected Tpico - T end interval and QT interval in patients with Chagas disease, chronic phase and normal electrocardiogram

FUNDAMENTO TEÓRICO: A doença de Chagas, uma das doenças parasitárias mais importantes no mundo atual carece de elementos que nos proporcionem prever quais pacientes evoluirão para a forma cardíaca e quais permanecerão na forma indeterminada. OBJETIVO: Avaliar a relação entre a evolução eletrocardiográfica e os intervalos Tpico-T fim e QT, ambos corrigidos para a frequência cardíaca, em pacientes chagásicos previamente com eletrocardiograma normal. CASUÍSTICA E MÉTODO: Avaliou-se retrospectivamente pacientes chagásicos admitidos na Instituição em 2002 ou antes, tendo eletrocardiograma normal na primeira consulta. Nas derivações V2 e V5 foi medido o intervalo Tpico-Tfim e QT em milessegundos, sendo a média destes valores corrigida para a frequência cardíaca (fórmula de Bazett). Analisou-se a relação destes valores com a evolução eletrocardiográfica destes pacientes, tendo em vista o sexo, o tempo de evolução e se receberam benzonidazol ou não. RESULTADOS: Em análise univariada, o intervalo Tpico-Tfim corrigido não mostrou significância estatística entre pacientes que mantiveram eletrocardiograma normal ou alterado (p: 0,7886). O intervalo QT corrigido (p: 0,0383), o tratamento com benzonidazol (p: 0,0001) e o tempo de evolução (p: 0,000) mostraram significância quanto à relação com a manutenção do eletrocardiograma normal ou não. Na avaliação multivariada, o tratamento com benzonidazol (OR: 0,260 - p: 0,0003), o intervalo QTc (OR: 1,009 - p: 0,0317) e o tempo de evolução (OR: 1,091 ­ p: 0,0001) se mostraram variáveis independentes para a manutenção do eletrocardiograma normal ou não. Quando se acrescentou as variáveis idade e sexo, os resultados se mantiveram. CONCLUSÃO: Em pacientes chagásicos com eletrocardiograma normal, o intervalo Tpico-T fim não se mostrou preditor da evolução eletrocardiográfica. O tratamento com benzonidazol favoreceu à manutenção do eletrocardiograma normal. A cada unidade de aumento do intervalo QTc houve aumento de 0,9% de chances de alterações eletrocardiográficas e a cada ano de evolução as chances do eletrocardiograma se tornar alterado foram de 9,1%. (AU)

Avaliação prognóstica do intervalo Tpico ­ T fim e intervalo QT corrigidos, em pacientes com doença de chagas, fase crônica e eletrocardiograma normal / Prognostic evaluation of corrected Tpico - T end interval and QT interval in patients with Chagas disease, chronic phase and normal electrocardiogram

FUNDAMENTO TEÓRICO: A doença de Chagas, uma das doenças parasitárias mais importantes no mundo atual carece de elementos que nos proporcionem prever quais pacientes evoluirão para a forma cardíaca e quais permanecerão na forma indeterminada. OBJETIVO: Avaliar a relação entre a evolução eletrocardiográfica e os intervalos Tpico-Tfim e QT, ambos corrigidos para a frequência cardíaca, em pacientes chagásicos previamente com eletrocardiograma normal. CASUÍSTICA E MÉTODO: Avaliou-se retrospectivamente pacientes chagásicos admitidos na Instituição em 2002 ou antes, tendo eletrocardiograma normal na primeira consulta. Nas derivações V2 e V5 foi medido o intervalo Tpico-Tfim e QT em milessegundos, sendo a média destes valores corrigida para a frequência cardíaca (fórmula de Bazett). Analisou-se a relação destes valores com a evolução eletrocardiográfica destes pacientes, tendo em vista o sexo, o tempo de evolução e se receberam benzonidazol ou não. RESULTADOS: Em análise univariada, o intervalo Tpico-Tfim corrigido não mostrou significância estatística entre pacientes que mantiveram eletrocardiograma normal ou alterado (p: 0,7886). O intervalo QTcorrigido (p: 0,0383), o tratamento com benzonidazol (p: 0,0001) e o tempo de evolução (p: 0,000) mostraram significância quanto à relação com a manutenção do eletrocardiograma normal ou não. Na avaliação multivariada, o tratamento com benzonidazol (OR: 0,260 - p: 0,0003), o intervalo QTc (OR: 1,009 - p: 0,0317) e o tempo de evolução (OR: 1,091 ­ p: 0,0001) se mostraram variáveis independentes para a manutenção do eletrocardiograma normal ou não. Quando se acrescentou as variáveis idade e sexo, os resultados se mantiveram (tabelas I e II). CONCLUSÃO: Em pacientes chagásicos com eletrocardiograma normal, o intervalo Tpico-Tfim não se mostrou preditor da evolução eletrocardiográfica. O tratamento com benzonidazol favoreceu à manutenção do eletrocardiograma normal. A cada unidade de aumento do intervalo QTc houve aumento de 0,9% de chances de alterações eletrocardiográficas e a cada ano de evolução as chances do eletrocardiograma se tornar alterado foram de 9,1%. (AU)

Doença de Chagas nas mulheres em idade fértil da Comunidade Hispana Armênia em São Paulo / Chagas disease in women of childbearing age from the Armenian Hispana Community in São Paulo

INTRODUÇÃO: Programas de controle da transmissão vetorial e transfusional da doença de Chagas alcançaram sucesso no Brasil, Chile e Uruguai, porém, a transmissão congênita vem demonstrando importância crescente e pode ocorrer em 71% dos recém-nascidos de mães com infecção aguda durante a gravidez e em 1,6% na fase crônica de doença. A migração de mulheres em idade fértil de outros países da América Latina para o Brasil pode aumentar a incidência da transmissão vertical no País, justificando a necessidade de acompanhamento médico adequado em mulheres grávidas e tratamento imediato de crianças com infecção congênita, já que são altas as taxas de cura. Os objetivos do estudo foram estimar a prevalência da doença em mulheres com idade fértil da Comunidade Hispana Armênia, informar e esclarecer dúvidas a respeito da doença. MÉTODO: Depois de realizada a coleta de 5 ml de sangue, em tubo ativador de coágulo com gel separador, de cada voluntária, o diagnóstico da doença de Chagas foi estabelecido por dois testes sorológicos de imunofluorescência indireta e um teste imunoenzimático - ELISA indireto. RESULTADOS: Das 84 coletas realizadas, quatro resultados foram excluídos, por serem de mulheres menopausadas. De 80 amostras analisadas, duas apresentaram resultado positivo para doença de Chagas. CONCLUSÃO: Conclui-se que na comunidade estudada, a preocupação com a transmissão vertical da doença de Chagas é baixa, o que não exclui a necessidade de implementar programas de saúde pública que contemplem orientação, esclarecimentos contínuos, a triagem de rotina em gestantes e recém-nascidos, assegurando assim, a detecção e tratamento de casos agudos e crônicos. (AU)

Evaluation of in vitro anti-Trypanosoma cruzi activity of medications benznidazole, amiodarone hydrochloride, and their combination.

INTRODUCTION: Approximately seven to eight million people worldwide have Chagas disease. In Brazil, benznidazole is the most commonly used active drug against Trypanosoma cruzi; however, its efficacy is limited, and side effects are frequent. Recent studies suggest that amiodarone may be beneficial in the treatment of this disease, by exerting anti-T. cruzi action. This study evaluated changes in T. cruzi cell count in in vitro cultures subjected to different doses of benznidazole, amiodarone, and their combination. METHODS: T. cruzi (Y strain) cultures containing approximately 100,000 cells were treated with either 100mg, 50mg, 25mg, 12.5mg, or 10mg of benznidazole, amiodarone, or their combination. On the 4th day, cell count was compared to the baseline data. RESULTS: On the 4th day, no parasites were observed in any of the treated cultures. CONCLUSIONS: Benznidazole and amiodarone were equally effective in eliminating T. cruzi in culture. The combination of the two drugs was also equally effective, but our data cannot demonstrate synergism, as similar results were obtained when the drugs were tested individually or in combination. It is suggested that this study be repeated with other T. cruzi strains to determine whether similar results can be obtained again.

Evaluation of in vitro anti-Trypanosoma cruzi activity of medications benznidazole, amiodarone hydrochloride, and their combination

Abstract INTRODUCTION: Approximately seven to eight million people worldwide have Chagas disease. In Brazil, benznidazole is the most commonly used active drug against Trypanosoma cruzi; however, its efficacy is limited, and side effects are frequent. Recent studies suggest that amiodarone may be beneficial in the treatment of this disease, by exerting anti-T. cruzi action. This study evaluated changes in T. cruzi cell count in in vitro cultures subjected to different doses of benznidazole, amiodarone, and their combination. METHODS: T. cruzi (Y strain) cultures containing approximately 100,000 cells were treated with either 100mg, 50mg, 25mg, 12.5mg, or 10mg of benznidazole, amiodarone, or their combination. On the 4th day, cell count was compared to the baseline data. RESULTS: On the 4th day, no parasites were observed in any of the treated cultures. CONCLUSIONS: Benznidazole and amiodarone were equally effective in eliminating T. cruzi in culture. The combination of the two drugs was also equally effective, but our data cannot demonstrate synergism, as similar results were obtained when the drugs were tested individually or in combination. It is suggested that this study be repeated with other T. cruzi strains to determine whether similar results can be obtained again.

Molecular detection of Trypanosoma cruzi in acai pulp and sugarcane juice.

Chagas disease, caused by Trypanosoma cruzi affects about 6-8 million people worldwide. Although transmission by triatomine insects has been controlled, other means of transmission maintain the infection. These forms of transmission are responsible for introducing Chagas disease in other non-endemic countries of the world. Thus, Chagas disease, nowadays is a worldwide health problem. In Brazil, acai pulp and sugarcane juice have been associated with Chagas disease outbreaks. The difficulties in isolation of the parasite from foods are hampering source tracking which could allow the confirmation of an implicated food commodity in these outbreak investigations. To address this scientific gap, we evaluated the performance of real-time PCR (qPCR) for detecting T. cruzi in acai pulp and sugarcane juice. All experiments were performed with acai pulp and sugarcane juice samples contaminated with different concentrations of T. cruzi. In assays with qPCR, the results showed that the ideal procedure for T. cruzi identification in acai pulp and sugarcane juice consisted of: i. centrifugation; ii. DNA extraction with a commercial kit for stool matrix; and iii. qPCR using a specific molecular marker for T. cruzi. The seeding in LIT medium of experimentally contaminated foods was effective in detecting the parasitic load by qPCR. The efficacy of qPCR was also verified testing food samples crushed with infected Triatomines. In conclusion, this methodology can be used to perform rapid diagnosis in outbreaks, facilitating measures in disease control.

Molecular detection of Trypanosoma cruzi in acai pulp and sugarcane juice

Chagas disease, caused by Trypanosoma cruzi affects about 6-8 million people worldwide. Although transmission by triatomine insects has been controlled, other means of transmission maintain the infection. These forms of transmission are responsible for introducing Chagas disease in other non-endemic countries of the world. Thus, Chagas disease, nowadays is a worldwide health problem. In Brazil, acai pulp and sugarcane juice have been associated with Chagas disease outbreaks. The difficulties in isolation of the parasite from foods are hampering source tracking which could allow the confirmation of an implicated food commodity in these outbreak investigations. To address this scientific gap, we evaluated the performance of real-time PCR (qPCR) for detecting T. cruzi in acai pulp and sugarcane juice. All experiments were performed with acai pulp and sugarcane juice samples contaminated with different concentrations of T. cruzi. In assays with qPCR, the results showed that the ideal procedure for T. cruzi identification in acai pulp and sugarcane juice consisted of: i. centrifugation; ii. DNA extraction with a commercial kit for stool matrix; and iii. qPCR using a specific molecular marker for T. cruzi. The seeding in LIT medium of experimentally contaminated foods was effective in detecting the parasitic load by qPCR. The efficacy of qPCR was also verified testing food samples crushed with infected Triatomines. In conclusion, this methodology can be used to perform rapid diagnosis in outbreaks, facilitating measures in disease control.

Evaluation of Parasiticide Treatment with Benznidazol in the Electrocardiographic, Clinical, and Serological Evolution of Chagas Disease.

INTRODUCTION: Chagas disease is one of the most important endemic parasitic diseases in Latin America. In its chronic phase, progression to cardiomyopathy has high morbidity and mortality. The persistence of a normal electrocardiogram (ECG) provides a similar prognosis to that of a non-diseased population. Benznidazole (BNZ) is the only drug with trypanocidal action available in Brazil. MATERIALS/METHODS/RESULTS: A group of 310 patients with chronic Chagas disease who had normal ECGs at the first medical visit performed before 2002 were included. There were 263 patients treated with BNZ and 47 untreated. The follow-up period was 19.59 years. Univariate analyses showed that those treated were younger and predominantly male. As many as 79.08% of those treated and 46.81% of those untreated continued with normal electrocardiograms (p <0.0001). The occurrence of electrocardiographic abnormalities and relevant clinical events (heart failure, stroke, total mortality, and cardiovascular death) was less prevalent in treated patients (p <0.001, p: 0.022, p: 0.047 respectively). In multivariate analyses, the parasiticide treatment was an independent variable for persistence of a normal ECG pattern, which was an independent variable in the prevention of significant clinical events. The immunofluorescence titers decreased with the parasitological treatment. However, the small number of tests in untreated patients did not allow the correlation of the decrease of these titers with electrocardiographic alterations. CONCLUSION: These data suggest that treatment with benznidazole prevents the occurrence of electrocardiographic alterations. On the other hand, patients who develop ECG abnormalities present with more significant clinical events.

Evaluation of parasiticide treatment with benznidazol in the electrocardiographic, clinical, and serological evolution of Chagas Disease

Chagas disease is one of the most important endemic parasitic diseases in Latin America. In its chronic phase, progression to cardiomyopathy has high morbidity and mortality. The persistence of a normal electrocardiogram (ECG) provides a similar prognosis to that of a non-diseased population. Benznidazole (BNZ) is the only drug with trypanocidal action available in Brazil. MATERIALS/METHODS/RESULTS: A group of 310 patients with chronic Chagas disease who had normal ECGs at the first medical visit performed before 2002 were included. There were 263 patients treated with BNZ and 47 untreated. The follow-up period was 19.59 years. Univariate analyses showed that those treated were younger and predominantly male. As many as 79.08% of those treated and 46.81% of those untreated continued with normal electrocardiograms (p <0.0001)...

Influence sexual dimorphism on the persistence of blood parasites in infected Calomys callosus / Influence sexual dimorphism on the persistence of blood parasites in infected Calomys callosus

Gender has long been known to be a contributory factor in the incidence and progression of disorders associated with immune system disregulation. The aims of this experiment were to verify the influences of sexual dimorphism on the persistence of blood parasites out of

the acute phase of infection. Male and female Calomys callosus were separated and infected with two strains of Trypanosoma cruzi, and let age until 120 days. Xenogiagnostic, culture of organs and blood, histopathology and lytic antibody percentages were evaluated on late

chronic phase. Xenodiagnosis, hemoculture and lytic antibody percentages were positive from 45 until 120 days. For both strains in adrenal

and heart, amastigote burdens were present until 45 days, scarcely found on 60 days and absent on 120 days. Steroid hormones, although having a protective role, does not enable animals to get completely rid of the infection. Even without showing apparent signs

of pathological unbalance, parasite persists, hidden throughout the host’s body.

Influence sexual dimorphism on the persistence of blood parasites in infected Calomys callosus.

Gender has long been known to be a contributory factor in the incidence and progression of disorders associated with immune system disregulation. The aims of this experiment were to verify the influences of sexual dimorphism on the persistence of blood parasites out of the acute phase of infection. Male and female Calomys callosus were separated and infected with two strains of Trypanosoma cruzi, and let age until 120 days. Xenogiagnostic, culture of organs and blood, histopathology and lytic antibody percentages were evaluated on late chronic phase. Xenodiagnosis, hemoculture and lytic antibody percentages were positive from 45 until 120 days. For both strains in adrenal and heart, amastigote burdens were present until 45 days, scarcely found on 60 days and absent on 120 days. Steroid hormones, although having a protective role, does not enable animals to get completely rid of the infection. Even without showing apparent signs of pathological unbalance, parasites persists, hidden throughout the host's body.