A morning with Louis Pasteur: a short history of the "clean hands".

This article examines the story of Louis Pasteur from the point of view of a classic movie presented at the Weekly Seminars of the "Oswaldo Cruz Institute", at the end of the 2017 activities. Although very old, the movie The Story of Louis Pasteur (Warner Bros., 1936) inspired spectators and gave rise to an energetic debate that led the authors to decide for publishing the comments of the Seminar Coordinator, the guest commentator and the audience. The movie communicates to the public the legacy of one of the greatest precursors of the public health history using also fictional characters. The article presents the reliable passages in Pasteur's biography and the fictional ones, without disrespecting the production of the creators of cinematographic work. The major merit of the movie, one of the first steps towards the policy of scientific diffusion, is to disclose the importance of vaccines and hand hygiene to prevent infectious diseases. The authors argue that the film-maker impeccably captured the scientist's tenacity in the relentless search for discoveries and Pasteur's idea that only persistent work can lead to rewarding results, remembering that the context created by previous researchers enabled Pasteur to establish new paradigms. Finally, the authors cite movie passages illustrating realities that are still in force: (i) the inertial resistance of science to new paradigms, illustrated by the medical-scientific community opposing to simple practices proposition, such as washing hands and boiling instruments, and (ii) the excessive confidence, and even arrogance, of some specialists, instead of serenity and humility that arise from committed study and accumulated knowledge.

A morning with Louis Pasteur: a short history of the "clean hands" / Uma manhã com Louis Pasteur: uma breve história das "mãos limpas" / Una mañana con Louis Pasteur: una pequeña historia sobre las "manos limpias"

Abstract: This article examines the story of Louis Pasteur from the point of view of a classic movie presented at the Weekly Seminars of the "Oswaldo Cruz Institute", at the end of the 2017 activities. Although very old, the movie The Story of Louis Pasteur (Warner Bros., 1936) inspired spectators and gave rise to an energetic debate that led the authors to decide for publishing the comments of the Seminar Coordinator, the guest commentator and the audience. The movie communicates to the public the legacy of one of the greatest precursors of the public health history using also fictional characters. The article presents the reliable passages in Pasteur's biography and the fictional ones, without disrespecting the production of the creators of cinematographic work. The major merit of the movie, one of the first steps towards the policy of scientific diffusion, is to disclose the importance of vaccines and hand hygiene to prevent infectious diseases. The authors argue that the film-maker impeccably captured the scientist's tenacity in the relentless search for discoveries and Pasteur's idea that only persistent work can lead to rewarding results, remembering that the context created by previous researchers enabled Pasteur to establish new paradigms. Finally, the authors cite movie passages illustrating realities that are still in force: (i) the inertial resistance of science to new paradigms, illustrated by the medical-scientific community opposing to simple practices proposition, such as washing hands and boiling instruments, and (ii) the excessive confidence, and even arrogance, of some specialists, instead of serenity and humility that arise from committed study and accumulated knowledge.

Resumo: O artigo examina a história de Louis Pasteur do ponto de vista de um filme clássico que foi projetado durante os Seminários Semanais do Instituto Oswaldo Cruz no final das atividades de 2017. Embora bastante antigo, o filme The Story of Louis Pasteur (Warner Bros., 1936) inspirou os espectadores e deu lugar a um debate animado que levou os autores a decidir publicar os comentários da Coordenadora dos Seminários, do debatedor convidado e do público. O filme incorpora personagens reais e fictícios para comunicar ao público o legado de um dos maiores precursores da história da saúde pública. O artigo destaca os episódios reais da biografia de Pasteur e também os ficcionais, sem desmerecer o trabalho dos criadores cinematográficos. O principal mérito do filme, um dos primeiros passos de uma política de divulgação científica, é de revelar a importância das vacinas e da higiene das mãos na prevenção das doenças infecciosas. Os autores argumentam que o cineasta retratou primorosamente a tenacidade do grande cientista na busca incansável por descobertas, além de sua ideia de que somente o trabalho persistente pode levar a resultados recompensadores, lembrando que o contexto criado por pesquisadores anteriores permitiu que Pasteur estabelecesse paradigmas novos. Finalmente, os autores citam trechos do filme que ilustram realidades que persistem até os dias de hoje: (i) a tendência da ciência de resistir, por inércia, aos paradigmas novos, exemplificada pela oposição da comunidade de ciência médica à proposição de práticas simples como a lavagem de mãos e a fervura de instrumentos e (ii) a confiança excessiva, e até arrogância, de alguns especialistas, em vez da serenidade e da humildade que nascem da pesquisa dedicada e do conhecimento acumulado.

Resumen: Este artículo examina la historia de Louis Pasteur desde el punto de vista de una película clásica, presentada en los Seminarios Semanales del Instituto Oswaldo Cruz, al final de las actividades de 2017. A pesar de ser muy antigua, la película The Story of Louis Pasteur (Warner Bros., 1936) inspiró a los espectadores y provocó un animado debate que condujo a los autores a la decisión de publicar los comentarios de la Coordinadora de los Seminarios, así como los del comentarista invitado y de la audiencia. Utilizando también personajes de ficción, la película transmitía al público el legado de uno de los más grandes precursores de la historia de la salud pública. El artículo señala pasajes fidedignos en la biografía de Pasteur y ficticios, sin menoscabar el trabajo de los creadores cinematográficos. El mayor mérito de la película, uno de los primeros pasos hacia la política de divulgación científica, es revelar la importancia de las vacunas y de la higiene de las manos para prevenir enfermedades infecciosas. Los autores enfatizan que el director de la película capturó impecablemente la tenacidad del científico en su búsqueda sin descanso de descubrimientos, así como su idea de que solo un trabajo persistente podía conducir a resultados gratificantes, recordando que el contexto creado por investigadores previos permitieron a Pasteur establecer nuevos paradigmas. Finalmente, los autores citan pasajes de la película que ilustran realidades todavía muy vigentes: (i) la resistencia por inercia de la ciencia a nuevos paradigmas, ilustrados por la oposición de la comunidad médico-científica hacia la propuesta de prácticas simples, tales como lavarse las manos y hervir los instrumentos, así como (ii) la excesiva confianza, e incluso arrogancia, de algunos especialistas, en lugar de la serenidad y humildad que surgen del estudio realizado y el conocimiento acumulado.

Multiple Approaches to Address Potential Risk Factors of Chagas Disease Transmission in Northeastern Brazil.

Chagas disease is one of the most significant systemic parasitosis in Latin America, caused by Trypanosoma cruzi, which is mainly transmitted by hematophagous insects, the triatomines. This research was carried out in both domestic and wild environments throughout a Northeastern rural locality. Triatomines were captured in both peridomicile and wild environments, obtaining 508 specimens of triatomines, of which 99.6% were Triatoma brasiliensis. Insects were captured in 10 (18.5%) peridomiciles with an average of 8.3 triatomines per residence. Triatoma brasiliensis nymphs and adults were found in six peridomiciles, generating a 11.1% colonization. No T. cruzi infection was detected in the 447 peridomestic insects analyzed. On the other hand, of the 55 sylvatic T. brasiliensis molecularly examined for T. cruzi, 12 (21%) were positive, all harboring T. cruzi I. The blood meal analysis by enzyme-linked immunosorbent assay from gut content revealed that both peridomestic and wild triatomine populations fed mainly on birds, refractory to the parasite, which may explain the null rate of natural infection prevalence in the domestic environment. However, infected triatomines for potential home infestation within the radius of insect dispersion capacity were registered in rock outcrops around the dwellings. Anthropogenic environmental influences are able to rapidly alter these scenarios. Therefore, to avoid disease transmission to humans, we recommend constant vector control combined with periodic serological surveillance. The associated methodology presented herein may serve as a model for early detections of risk factors for Chagas disease transmission in the Brazilian Northeast.

Genetic diversity of Giardia duodenalis circulating in three Brazilian biomes.

Giardia duodenalis has a wide genetic variety, and its characterization helps in the understanding of its transmission dynamics and in the development control strategies. This study aimed to assess the genetic diversity of G. duodenalis obtained in different Brazilian biomes and estimate their phylogenetic relationships. Three surveys including 944 participants were carried out in the municipalities of Russas (RSS, Caatinga semiarid biome), Santa Isabel do Rio Negro (SIRN, Amazon rainforest biome) and Nossa Senhora de Nazaré (NSN, Cerrado-Caatinga transition biome). G. duodenalis-positive fecal samples were submitted to amplification of gene fragments encoding ß-giardin (ßG, N = 71), glutamate dehydrogenase (GDH, N = 42), and triosephosphate isomerase (TPI, N = 27). Overall detection rates of assemblage A in G. duodenalis-positive samples through ßG, GDH and TPI were 22/71 (31%), 13/42 (31%), and 13/27 (48.1%), respectively. Concerning assemblage B, rates with distinct genetic markers were 49/71 (69%), 29/42 (69%), and 14/27 (51.9%), respectively. In the Amazon, assemblage B was more prevalent (77.8%, 71.8% and 65% through ßG, GDH and TPI, respectively), while in the Cerrado biome assemblage A predominated (50%, 66.6%, and 85.7%, through ßG, GDH and TPI, respectively). In Caatinga biome assemblage A also predominated (71.4%, through ßG). Thirty new sub-assemblages are described for assemblage B (24 ßG and six TPI), as well as three new sub-assemblages are described for assemblage A (one GDH and 2 TPI). Higher genetic diversity of assemblage B in the Amazon may be related to demographic concentration leading to a more complex transmission network within a poorer sanitation background. The high genetic divergence between assemblages A and B (5.5-6.3%) support the proposal of taxon separation in distinct species.

Giardia intestinalis infection associated with malnutrition in children living in northeastern Brazil.

INTRODUCTION: The present study aimed to determine the prevalence and factors associated with Giardia intestinalis infection, verifying its impact on the nutritional status of children in northeastern Brazil. METHODOLOGY: A cross-sectional study was conducted to obtain parasitological, sociodemographic, and anthropometric data in two municipalities in the states of Piauí and Ceará, northeastern Brazil. RESULTS: Prevalence of giardiasis was 55/511 (10.8%). G. intestinalis was more frequent in people living in poverty (30/209 [14.4%], p = 0.041), performing open evacuation (26/173 [15%], p = 0.034), and drinking rainwater stored in cisterns (9/56 [16.1%], p = 0.005). The proportion of stunting and being underweight in children infected with G. intestinalis was significantly higher than that in uninfected children (5/23 [21.7%] vs. 10/179 [5.6%], p = 0.017, OR = 4.69, 95% confidence interval [CI] = 1.44-15.25 and 5/23 [21.7%] vs. 13/179 [7.3%], p = 0.038, OR = 3.54, 95% CI = 1.13-11.09, respectively). Infection with G. intestinalis remained significantly associated with stunting and being underweight after adjustment for poverty, municipality, sex, and age in a logistic regression multivariate model. CONCLUSIONS: In rural areas in northeastern Brazil, giardiasis has acquired great public health importance in the soil-transmitted helminths control era, impacting the nutritional status of children and requiring new approaches to diagnosis and treatment and translational research that could generate applicable solutions at the community level.

Everybody loves sugar: first report of plant feeding in triatomines.

BACKGROUND: Triatomines, which are the vectors of Trypanosoma cruzi, have been considered to be exclusive blood feeders for more than 100 years, since the discovery of Chagas disease. METHODS: We offered artificial sugar meals to the laboratory model-insect Rhodnius prolixus, which is considered a strict haematophagous insect. We registered feeding by adding colorant to sugar meals. To assess putative phytophagy, fruits of the tomato Solanum lycopersicum were offered to R. prolixus and the presence of tomato DNA was assessed in the insects using PCR. We also assessed longevity, blood feeding and urine production of fruit-exposed triatomines and control insects. RESULTS: All instars of R. prolixus ingested sugar from artificial sugar meals in laboratory conditions. First instar R. prolixus ingested plant tissue from S. lycopersicum fruits, and this increased the amount of blood ingested and urine excreted. Decreased mortality was also observed after blood feeding. Exposure to S. lycopersicum increased longevity and reduced weight loss caused by desiccation. CONCLUSIONS: We describe here the first report of sugar feeding and phytophagy in a species that was considered to be a strict blood-feeder for over a century. We suggest that local plants might be not merely shelters for insects and vertebrate hosts as previously described, but may have a nutritional role for the maintenance of the triatomine vectors. The description of sugar and plant meals in triatomines opens new perspectives for the study and control of Chagas Disease.

Frequency and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, and Entamoeba hartmanni in the context of water scarcity in northeastern Brazil.

This study aimed to estimate the frequency, associated factors, and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, andEntamoeba hartmanni infections. We performed a survey (n = 213 subjects) to obtain parasitological, sanitation, and sociodemographic data. Faecal samples were processed through flotation and centrifugation methods.E. histolytica, E. dispar, E. moshkovskii, and E. hartmanni were identified by nested-polymerase chain reaction (PCR). The overall prevalence of infection was 22/213 (10.3%). The infection rate among subjects who drink rainwater collected from roofs in tanks was higher than the rate in subjects who drink desalinated water pumped from wells; similarly, the infection rate among subjects who practice open defecation was significantly higher than that of subjects with latrines. Out of the 22 samples positive for morphologically indistinguishableEntamoeba species, the differentiation by PCR was successful for 21. The species distribution was as follows: 57.1% to E. dispar, 23.8% to E. histolytica, 14.3% toE. histolytica and E. dispar, and 4.8% E. dispar and E. hartmanni. These data suggest a high prevalence of asymptomatic infection by the group of morphologically indistinguishable Entamoeba histolytica/dispar/moshkovskiicomplex and E. hartmanni species. In this context of water scarcity, the sanitary and socioenvironmental characteristics of the region appear to favour transmission.

Frequency and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, and Entamoeba hartmanni in the context of water scarcity in northeastern Brazil

This study aimed to estimate the frequency, associated factors, and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, andEntamoeba hartmanni infections. We performed a survey (n = 213 subjects) to obtain parasitological, sanitation, and sociodemographic data. Faecal samples were processed through flotation and centrifugation methods.E. histolytica, E. dispar, E. moshkovskii, and E. hartmanni were identified by nested-polymerase chain reaction (PCR). The overall prevalence of infection was 22/213 (10.3%). The infection rate among subjects who drink rainwater collected from roofs in tanks was higher than the rate in subjects who drink desalinated water pumped from wells; similarly, the infection rate among subjects who practice open defecation was significantly higher than that of subjects with latrines. Out of the 22 samples positive for morphologically indistinguishableEntamoeba species, the differentiation by PCR was successful for 21. The species distribution was as follows: 57.1% to E. dispar, 23.8% to E. histolytica, 14.3% toE. histolytica and E. dispar, and 4.8% E. dispar and E. hartmanni. These data suggest a high prevalence of asymptomatic infection by the group of morphologically indistinguishable Entamoeba histolytica/dispar/moshkovskiicomplex and E. hartmanni species. In this context of water scarcity, the sanitary and socioenvironmental characteristics of the region appear to favour transmission.

Phenotypic variability confirmed by nuclear ribosomal DNA suggests a possible natural hybrid zone of Triatoma brasiliensis species complex.

Triatoma brasiliensis macromelasoma occurs in Pernambuco state, Brazil, which is situated between the distribution areas of Triatoma brasiliensis brasiliensis (north) and Triatoma juazeirensis (south). T. b. macromelasoma displays greater variations in its chromatic phenotype than either T. b. brasiliensis or T. juazeirensis, and patterns reminiscent of one or the other. Experimental crosses from each of these members of the T. brasiliensis species complex generated fertile offspring suggesting that viable hybrids could be present in nature, despite their significant genetic distances. Considering the geographical position of occurrence of the T. b. macromelasoma (in Pernambuco) it was proposed to be an area capable of supporting natural hybridization between T. b. brasiliensis and T. juazeirensis. Since phenotypic variability is expected, this study investigated the existence of intermediate chromatic phenotypes for T. b. macromelasoma in various locations in areas between the T. b. brasiliensis and T. juazeirensis occurrences. Thirteen different color patterns were for the first time characterized and nine of those displayed intermediate phenotypes. Molecular analysis performed using ribosomal DNA intergenic region, grouped all within the T. brasiliensis complex. The intermediate chromatic phenotypes, molecular analysis and experimental crosses all support the distinction of a zone of hybridization that gave rise to the T. b. macromelasoma through homoploidal evolution.

Molecular Identification of Food Sources in Triatomines in the Brazilian Northeast: Roles of Goats and Rodents in Chagas Disease Epidemiology.

We used the gut contents of triatomines collected from rural areas of Ceará State, northeastern Brazil, to identify their putative hosts via vertebrate cytb gene sequencing. Successful direct sequencing was obtained for 48% of insects, comprising 50 Triatoma brasiliensis, 7 Triatoma pseudomaculata, and 1 Rhodnius nasutus. Basic local alignment search tool (BLAST) procedure revealed that domestic animals, such as chickens (Gallus gallus) and goats (Capra hircus), are the main food source, including in sylvatic environment. Native hosts were also detected in peridomestic environment such as reptiles (Tropidurus sp. and Iguana iguana) and the Galea spixii (Rodentia: Caviidae). The role of goats and Galea spixii in Chagas disease epidemiology calls for further studies, because these mammals likely link the sylvatic and domestic Trypanosoma cruzi cycles.

Characterization of the microbiota in the guts of Triatoma brasiliensis and Triatoma pseudomaculata infected by Trypanosoma cruzi in natural conditions using culture independent methods.

BACKGROUND: Chagas disease is caused by Trypanosoma cruzi, which is transmitted by triatomine vectors. The northeastern region of Brazil is endemic for Chagas disease and has the largest diversity of triatomine species. T. cruzi development in its triatomine vector depends on diverse factors, including the composition of bacterial gut microbiota. METHODS: We characterized the triatomines captured in the municipality of Russas (Ceará) by sequencing the cytochrome c oxidase subunit I (COI) gene. The composition of the bacterial community in the gut of peridomestic Triatoma brasiliensis and Triatoma pseudomaculata was investigated using culture independent methods based on the amplification of the 16S rRNA gene by polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), DNA fragment cloning, Sanger sequencing and 454 pyrosequencing. Additionally, we identified TcI and TcII types of T. cruzi by sequencing amplicons from the gut metagenomic DNA with primers for the mini-exon gene. RESULTS: Triatomines collected in the peridomestic ecotopes were diagnosed as T. pseudomaculata and T. brasiliensis by comparing their COI sequence with GenBank. The rate of infection by T. cruzi in adult triatomines reached 80% for T. pseudomaculata and 90% for T. brasiliensis. According to the DNA sequences from the DGGE bands, the triatomine gut microbiota was primarily composed of Proteobacteria and Actinobacteria. However, Firmicutes and Bacteroidetes were also detected, although in much lower proportions. Serratia was the main genus, as it was encountered in all samples analyzed by DGGE and 454 pyrosequencing. Members of Corynebacterinae, a suborder of the Actinomycetales, formed the next most important group. The cloning and sequencing of full-length 16S rRNA genes confirmed the presence of Serratia marcescens, Dietzia sp., Gordonia terrae, Corynebacterium stationis and Corynebacterium glutamicum. CONCLUSIONS: The study of the bacterial microbiota in the triatomine gut has gained increased attention because of the possible role it may play in the epidemiology of Chagas disease by competing with T. cruzi. Culture independent methods have shown that the bacterial composition of the microbiota in the guts of peridomestic triatomines is made up by only few bacterial species.

Autochthonous transmission of Chagas disease in Rio de Janeiro State, Brazil: a clinical and eco-epidemiological study.

BACKGROUND: After the control of the main modes of Chagas disease (CD) transmission in most endemic countries, it is important to identify the participation of native sylvatic vectors in CD transmission. Although CD is not considered endemic in Rio de Janeiro State (RJ), Brazil, we identified patients with CD born in RJ and investigated the possible autochthonous transmission in the state. METHODS: Patients born in RJ and followed in our institution between 1986 and 2011 were retrospectively analyzed. The cases identified as autochthonous transmission were submitted to epidemiological, clinical, serological, parasitological and molecular studies. Sectional field study with serological survey, research of sylvatic reservoirs and vectors was conducted in rural areas where patients were born. RESULTS: Among 1963 patients, 69 (3.5%) were born in RJ. From these, 15 (21.7%) were considered to have acquired the infection by autochthonous transmission. Cardiac form was the commonest form of presentation (60%). In rural areas in RJ northern region, sylvatic cycles of Trypanosoma cruzi and domestic invasion by Triatoma vitticeps were identified, and CD prevalence among inhabitants was 0.74%.TcI genotype was identified in sylvatic reservoirs and vectors. The genotype (mixed infection TcI/TcVI) could be identified in one of the autochthonous cases. CONCLUSIONS: The autochthonous vectorial transmission of CD occurs in RJ, probably due to wild cycles of T. cruzi and sylvatic vectors, such as T. vitticeps. Therefore, the health authorities should evaluate if RJ should be included in the original endemic area of CD and CD should be included in the diagnostic work out of cardiomyopathy of patients born in RJ. Moreover, control and educational measures should be put into place in the risk areas.

An entomoepidemiological investigation of Chagas disease in the state of Ceará, Northeast Region of Brazil.

The seroprevalence of Chagas disease in humans and the presence of triatomines were investigated in a rural locality in the State of Ceará, Brazil, an historically endemic region. Approximately 80% of the surveyed residents agreed to undergo serological tests. Intradomestic and peridomestic environments were searched for triatomines in both the dry and rainy seasons. The prevalence rate of Chagas disease was 1.2% and the majority of individuals confirmed with the disease over 50 years of age. A total of 761 specimens of triatomines were captured, most of which were from colonies composed of nymphs and adult bugs, and the majority of specimens were obtained in the dry season. Triatoma brasiliensis was the predominant species. Analysis using light microscopy revealed that 28.6% of the insects were Trypanosoma cruzi positive. Results suggest that peridomestic man-made structures, such as animal shelters, improper storage of timber and uninhabited dwellings contribute to the high rate of triatomine infestation in the area.

An entomoepidemiological investigation of Chagas disease in the state of Ceará, Northeast Region of Brazil / Investigação entomoepidemiológica da doença de Chagas no Estado do Ceará, Nordeste do Brasil / Investigación entomoepidemiológica de la enfermedad de Chagas en el estado de Ceará, noreste de Brasil

The seroprevalence of Chagas disease in humans and the presence of triatomines were investigated in a rural locality in the State of Ceará, Brazil, an historically endemic region. Approximately 80% of the surveyed residents agreed to undergo serological tests. Intradomestic and peridomestic environments were searched for triatomines in both the dry and rainy seasons. The prevalence rate of Chagas disease was 1.2% and the majority of individuals confirmed with the disease over 50 years of age. A total of 761 specimens of triatomines were captured, most of which were from colonies composed of nymphs and adult bugs, and the majority of specimens were obtained in the dry season. Triatoma brasiliensis was the predominant species. Analysis using light microscopy revealed that 28.6% of the insects were Trypanosoma cruzi positive. Results suggest that peridomestic man-made structures, such as animal shelters, improper storage of timber and uninhabited dwellings contribute to the high rate of triatomine infestation in the area.

A presença de triatomíneos e a soroprevalência da infecção chagásica humana foram investigadas em uma localidade rural do Estado do Ceará, Brasil, região endêmica histórica da doença de Chagas. Os triatomíneos foram pesquisados nos domicílios, nos períodos de seca e de chuva. A soroprevalência encontrada no período foi de 1,2%, maior entre pessoas com mais de 50 anos de idade, e sem diferença significativa em relação ao sexo. Foram capturados 761 espécimes de triatomíneos, em colônias compostas de ninfas e insetos adultos, com predominância de Triatoma brasiliensis, especialmente no período de seca. O exame do conteúdo intestinal dos insetos revelou que 28,6% eram Trypanosoma cruzi positivos. Os resultados sugerem que os abrigos de animais e armazenamento inadequado de materiais no peridomicílio, particularmente amontoados de madeiras, bem como moradias desabitadas, contribuem para o alto índice de infestação de triatomíneos na localidade.

Se investigó la presencia de insectos y seroprevalencia en la infección humana por Trypanosoma cruzi, en un área rural del estado de Ceará, Brasil, región endémica histórica de la enfermedad Chagas. Se informó sobre la presencia de insectos en los hogares durante los períodos de sequía y lluvia. La seroprevalencia en el período fue un 1,2%, mayor entre las personas de más de 50 años de edad, y no hubo diferencias significativas en relación al sexo. Se capturaron 761 ejemplares de insectos en colonias compuestas por ninfas y adultos de insectos, principalmente, Triatoma brasiliensis, especialmente durante la estación seca. El examen del contenido intestinal de los insectos reveló que un 28,6% eran T. cruzi positivo. Los resultados sugieren que los refugios de animales y el almacenamiento inadecuado de materiales fuera del hogar, especialmente pilas de madera y casas deshabitadas, contribuyen a la alta tasa de infestación de insectos en la localidad.

Exploring the Role of Rhodtestolin, A Cardio-Inhibitor from the Testes of Rhodnius prolixus, in Relation to the Structure and Function of Reproductive Organs in Insect Vectors of Chagas Disease.

Rhodtestolin is a cardio-inhibitor that was first discovered in testes extracts of the blood-feeding insect, Rhodnius prolixus. Its role in reproduction remains unconfirmed, but if delivered to the female during spermatophore formation, it may serve to calm the female and/or relax the vaginal muscles to facilitate delivery and storage of the spermatophore. We describe here the anatomy of reproductive organs in R. prolixus and show that rhodtestolin is present in a low-molecular weight fraction of testes extracts separated by gel filtration, as well as in spermatophores delivered to the female during spermatophore formation. We also report that a rhodtestolin-like factor is present in the testes of R. brethesi, Triatoma dimidiata, T. klugi and Nesotriatoma bruneri, other Reduviidae, which are vectors of Chagas disease. Male secretions in insects are known to modify female behavior after copulation, and the presence of rhodtestolin in several genera of Reduviidae suggests that it plays an important role in reproductive success. Determining this role could lead to developing additional population control strategies for these bugs.

Doença de Chagas: situação vetorial no município de Limoeiro do Norte – CE, no período de 2006 a 2009 / Chagas Disease: vector status in the city of Limoeiro do Norte – CE, during the period from 2006 to 2009

Entre as doenças transmitidas por vetores no Brasil, a doença de Chagas é uma das mais importantes como problemas de saúde pública. Neste estudo foi estimada a prevalência de triatomíneos infectados por Trypanosoma cruzi no município de Limoeiro do Norte-CE, por meio de consulta aos arquivos do Programa de Controle da Doença de Chagas deste Distrito Sanitário no período de 2006 a 2009. Na análise foram incluídos os triatomíneos adultos e ninfas, mortos ou vivos, capturados durante o período investigado. Durante os quatro anos de estudo foram capturados 2.863 exemplares de triatomíneos, ninfase adultos distribuídos no intradomicílio e peridomicílio; e examinados 2.756 (96,3 %) exemplares. O índice de infecção dos triatomíneos foi de 7,9 % nos quatros anos de estudo. O número de ninfas capturadas foi cerca de três vezes maior que o número de adultos; no entanto, o número de ninfas infectadas pelo flagelado (5,30 %) foi inferior aos adultos (15,85 %). O distrito de Limoeiro do Norte apresenta alto risco de transmissão para doença de Chagas e, portanto, sugere-se que as ações de controle sejam intensificadas neste município para evitar a ocorrência de transmissão da doença e dispersão do vetor.

In Brazil, among the endemic vector-borne diseases, Chagas disease has been considered one of the mostimportant public health problems. The present study aimed at estimating the prevalence of triatominesinfected with Trypanosoma cruzi in the city of Limoeiro do Norte, CE, Brazil. This investigation was conductedby consulting the archives of the Chagas disease control program of the Health District of Limoeiro do Nortefor the period from 2006 to 2009. The study included dead or alive, adults and nymphs triatomines, whichwhere captured during the investigated period. During the four-year study, 2,863 specimens of nymphs andadults insects were captured from the domestic and peridomestic areas; 2,756 were examined, representing96.3 % of collected specimens. The insect infection rate was 7.9 % in four-year study. The number of capturednymphs was nearly three times higher than the number of adults; however the number of nymphs infectedwith the flagellate (5.30 %) was lower than the number of adults form (15.85 %). Limoeiro do Norte showeda high risk for Chagas disease transmission, suggesting that the control actions should be intensified in thiscity to prevent the disease transmission and the vector spread.

Cultivation-independent methods reveal differences among bacterial gut microbiota in triatomine vectors of Chagas disease.

BACKGROUND: Chagas disease is a trypanosomiasis whose agent is the protozoan parasite Trypanosoma cruzi, which is transmitted to humans by hematophagous bugs known as triatomines. Even though insecticide treatments allow effective control of these bugs in most Latin American countries where Chagas disease is endemic, the disease still affects a large proportion of the population of South America. The features of the disease in humans have been extensively studied, and the genome of the parasite has been sequenced, but no effective drug is yet available to treat Chagas disease. The digestive tract of the insect vectors in which T. cruzi develops has been much less well investigated than blood from its human hosts and constitutes a dynamic environment with very different conditions. Thus, we investigated the composition of the predominant bacterial species of the microbiota in insect vectors from Rhodnius, Triatoma, Panstrongylus and Dipetalogaster genera. METHODOLOGY/PRINCIPAL FINDINGS: Microbiota of triatomine guts were investigated using cultivation-independent methods, i.e., phylogenetic analysis of 16s rDNA using denaturing gradient gel electrophoresis (DGGE) and cloned-based sequencing. The Chao index showed that the diversity of bacterial species in triatomine guts is low, comprising fewer than 20 predominant species, and that these species vary between insect species. The analyses showed that Serratia predominates in Rhodnius, Arsenophonus predominates in Triatoma and Panstrongylus, while Candidatus Rohrkolberia predominates in Dipetalogaster. CONCLUSIONS/SIGNIFICANCE: The microbiota of triatomine guts represents one of the factors that may interfere with T. cruzi transmission and virulence in humans. The knowledge of its composition according to insect species is important for designing measures of biological control for T. cruzi. We found that the predominant species of the bacterial microbiota in triatomines form a group of low complexity whose structure differs according to the vector genus.

Eco-epidemiology of Chagas disease in northeastern Brazil: Triatoma brasiliensis, T. pseudomaculata and Rhodnius nasutus in the sylvatic, peridomestic and domestic environments.

An entomological survey was carried out in four rural localities situated in the state of Ceará, assessing Chagas disease seroprevalence in man, focusing on the presence of vectors in natural foci contiguous to the domestic and peridomestic environments. Fifty-three Triatoma brasiliensis, nine T. pseudomaculata and 71 Rhodnius nasutus were collected in their natural habitats as far as 10 m from the houses, and 663, 59 and 8 respectively were captured in peridomestic artificial structures, adjacent to the houses, including henhouses, pigpens, corrals, perches and piles of bricks, tiles and wood. Within the households, 37 T. brasiliensis, one specimen of T. pseudomaculata and one of R. nasutus were captured. Overall, Trypanosoma cruzi infection rates were 2.3% for T. brasiliensis and 11.3% for R. nasutus. Despite that the seroprevalence survey in man did not reveal positive results using two serological techniques, natural triatomine habitats are juxtaposed to man-made artificial ecotopes, resulting in overlapping habitats. The contiguity between natural ecotopes and human dwellings increases the interaction between vectors and humans, challenging continuous surveillance and vector control efforts.

In vitro trypanocidal activity of DB745B and other novel arylimidamides against Trypanosoma cruzi.

OBJECTIVES: As part of a search for new therapeutic opportunities to treat chagasic patients, in vitro efficacy studies were performed to characterize the activity of five novel arylimidamides (AIAs) against Trypanosoma cruzi. METHODS: The trypanocidal effect against T. cruzi was evaluated by light microscopy through the determination of IC50 values. Cytotoxicity was determined by MTT assays against mouse cardiomyocytes. RESULTS: Our data demonstrated the trypanocidal efficacy of these new compounds against bloodstream trypomastigotes and intracellular amastigotes, exhibiting IC50 values ranging from 0.015 to 2.5 and 0.02 to0.2 µM, respectively. One of the compounds, DB745B, was also highly active against a broad panel of isolates, including those naturally resistant to benznidazole. DB745B showed higher in vitro efficacy than the reference drugs used to treat patients (benznidazole IC50= 12.94 µM) and to prevent blood bank infection (gentian violet IC50= 30.6 µM). CONCLUSIONS: AIAs represent promising new chemical entities against T. cruzi and are also potential trypanocidal agents to prevent transfusion-associated Chagas' disease.

Ecology of Triatoma brasiliensis in northeastern Brazil: seasonal distribution, feeding resources, and Trypanosoma cruzi infection in a sylvatic population.

We assessed some ecological parameters of Triatoma brasiliensis in rock piles in the state of Ceará during the rainy and dry seasons. The greatest density was in April (median = 12.5 triatomines/site). The greatest abundance was in December, when the insects were more dispersed and the density per site was lower (6 triatomines/site). The nutritional status of females and 5th instar nymphs was increased in July. The rate of T. cruzi infection reached its highest peak in July (10.9%). ELISA revealed that the principal food sources were birds (33.1%), followed by armadillos (18.8%). Food sources were more frequently identified during the rainy season. T. brasiliensis specimens collected in the drought tended to: i) present lower rates of T. cruzi infection and gut content reactivity to tested antisera, ii) have a poorer nutritional status, iii) exhibit lower fecundity, iv) be more dispersed among the studied collection sites, and v) be more abundant and easily collected in the surface of the rocks, possibly reflecting an increased searching for blood meals. Such findings underscore epidemiological concerns and allow inferences about the season when triatomines can more frequently invade the peridomestic environment in search of food and recolonize artificial structures.