Studies in search of a suitable experimental insect model for xenodiagnosis of hosts with Chagas' disease: 3 - On the interaction of vector species and parasite strain in the reaction of bugs to infection by Trypanosoma cruzi

The reaction of nine vector species of Chagas' disease to infection by seven different Trypanosoma cruzi strains; Berenice, Y, FL, CL, S. Felipe, Colombiana and Gávea, are examined and compared. On the basis of the insects' ability to establish and maintain the infection, vector species could be divided into two distinct groups which differ in their reaction to an acute infection by T. cruzi. While the proportion of positive bugs was found to be low in Triatoma infestans and Triatoma dimidiata it was high, ranging from 96.9% to 100% in the group of wild (Rhodnius neglectus, Triatoma rubrovaria)and essentially sylvatic vectors in process of adaptation to human dwellings, maintained under control following successful insecticidal elimination of Triatoma infestans (Panstrongylus megistus, Triatoma sordida and Triatoma pseudomaculata). An intermediate position is held by Triatoma brasiliensis and Rhodnius prolixus. This latter has been found to interchange between domestic and sylvatic environments. The most important finding is the strikingly good reaction between each species of the sylvatic bugs and practically all T. cruzi strains herein studied, thus indicating that the factors responsible for the excellent reaction of P.megistus to infection by Y strain, as previously reported also come into operation in the reaction of the same vector species to acute infections by five of the remaining T.cruzi strains. Comparison or data reported by other investigators with those herein described form the basis of the discussion of Dipetalogaster maximus as regards its superiority as a xenodiagnostic agent.

Studies in search of a suitable experimental insect model for xenodiagnosis of hosts with Chagas' disease: 4 - The reflection of parasite stock in the responsiveness of different vector species to chronic infection with different Trypanosoma cruzi stocks

Previous studies (1982,1987) have emphasized the superiority of sylvatic vector species over domestic species as xenodiagnostic agents in testing hosts with acute or chronic infections by T. cruzi "Y" stock. The present study, which is unique in that it contains data on both infectivity rates produced by the same stock in 11 different vector species and also the reaction of the same vector species to seven different parasite stocks, establishes the general validity of linking efficiency of xenodiagnosis to the biotope of its agent. For example, infectivity rates produced by "São Felipe" stock varied from 82.5% to 98.3% in sylvatic vectors but decreased to 42.5% to 71.3% in domestic species. "Colombiana" stock produced in the same sylvatic vectors infectivity rates ranging from 12.5% to 45%. These shrank to 5%-22.5% in domestic bugs. The functional role of the biotope in the vector-parasite interaction has not been eluddated. But since this phenomenon has been observed to be stable and easy to reproduce, it leads us to believe that the results obtained are valid. Data presented also provide increasing evidence that the infectivity rates exhibited by bugs from xenodiagnosis in chronic hosts, are parasite stock specific. For example, infectivity rates produced by "Berenice", "Y", "FL" and "CL" varied in R. neglectus from 26.3% to 75%; in P. megistus from 56.3% to 83.8%; in T. sordida from 28.8% to 58.8% in T. pseudomaculata from 41.3% to 66.3% and in T. rubrovaria from 48.8% to 85%. Data from xenodiagnosis in the same hosts, carrying acute infections by the same parasite stocks, gave the five sylvatic vectors a positive rating of approximately 100%, thus suggesting that the heavy loads of parasites circulating in the acute hosts obscured the characteristic interspecific differences for the parasite stock. Nonetheless these latter were revealed in the same hosts with chronic infections stimulated by very low numbers of the same parasite stocks. Certain observations here described lead us to speculate as to the possibility of further results from other parasite stocks, allowing the association of the infectivity rates produced in bugs by different parasite stocks with the isoenzymic patterns revealed by these stocks.

Pesquisa da proteína 24 do vírus da imunodeficiência humana (HIV) nas fezes de triatomíneos alimentados em pacientes com aids

OBJETIVO: Verificar se pode haver eliminação da proteína p24, antígeno que é um dos marcadores da infecção pelo HIV, pelas fezes de triatomíneos. Foi avaliado o possível risco de contaminação por parte de profissionais que exercem atividades laboratoriais relacionadas aos triatomíneos, e também verificado o eventual mecanismo de disseminação do HIV. MÉTODO: Os triatomíneos (Triatoma infestans) alimentaram-se com sangue de 23 pacientes acometidos de AIDS e nos quais estava presente a p24. As fezes desses insetos foram examinadas 24 e 48 horas depois como tentativas de evidenciar a presença do antígeno. As pesquisas da p24 sempre ocorreram por meio de técnica imunoenzimática. RESULTADO E CONCLUSÃO: Em nenhuma das ocasiões sucedeu detecção da p24. De acordo com a metodologia adotada o objetivo pôde ser alcançado, no sentido de mostrar que a eliminação da p24 nunca aconteceu. Talvez outras formas de agir revelem fatos diferentes e subsidiem o que se conhece quanto aos riscos de veiculação do HIV.

Chagas disease in an area of recent occupation in Cochabamba, Bolivia

INTRODUCTION: A descriptive, entomological and seroepidemiological study on Chagas disease was conducted in a place of recent occupation on the outskirts of Cochabamba, Bolivia: Avaroa/Primer de Mayo (population:3,000), where the socio-economic level is low and no control measures have been made available. METHODS: The immunofluorescent antibody test (IFAT) was used for IgG and IgM anti-Trypanosoma cruzi antibodies in filter paper bloodspot eluates from 128 subjects (73 females, 55 males) selected by systematic sampling. Concerning each subject age, gender, birthplace, occupation, duration of residence and building materials used in their houses were recorded. Vectors were captured both in domestic and peridomestic environments. RESULTS: Seropositive, 12.5% (16/128): females, 15.1% (11/73); males, 9.1% (5/55). Average time of residence: 6.1 years for the whole population sample and 7.4 years for the seropositive subjects. Most houses had adobe walls (76.7% , n= 30), galvanized iron rooves (86.7%) and earthen floors (53.4%) 80% of the walls had crevices. One hundred forty seven specimens of Triatoma infestans were captured, of which 104 (70.7%) were domestic, and 1 peridomestic Triatoma sordida. Precipitin host identification: birds, 67.5%; humans, 27.8%; rodents, 11.9%; dogs, 8.7%; cats, 1.6%. House infestation and density indices were 53.3 and 493.0 respectively. We found 21 (14.3%) specimens of T. infestans infected with trypanosomes, 18 (85.7%) of which in domestic environments. DISCUSSION: The elements for the vector transmission of Chagas disease are present in Avaroa/Primer de Mayo and the ancient custom of keeping guinea pigs indoors adds to the risk of human infection. In neighboring Cochabamba, due to substandard quality control, contaminated blood transfusions are not infrequent, which further aggravates the spread of Chagas disease. Prompt action to check the transmission of this infection, involving additionally the congenital and transfusional modes of acquisition, is required.

Soroepidemiologia da tripanosomíase americana na região do litoral sul, São Paulo

OBJETIVO: Analisar dados de reações sorológicas realizadas em dois períodos, entre fins da década de 1960 e o ano de 1983, quando a sorologia passou a complementar a informação sobre triatomíneos vetores do Estado de São Paulo e, de 1984 até 1999, caracterizando o panorama atual da endemia nessa região. MÉTODOS: Foram realizadas análises soroepidemiológicas de reações, obtidas em inquéritos escolares (fins da década de 1960 e entre 1973 e 1983), em inquéritos populacionais em Cananéia, Iguape e Peruíbe (entre 1980 e 1982) e em moradores de unidades domiciliares. Os resultados da análise foram associados a informações obtidas sobre os vetores encontrados. RESULTADOS: Manteve-se a sororreatividade entre escolares em patamares baixos, porém constantes, ao longo dos anos 1973 a 1982, tornando-se nulos em 1983. A curva de freqüência de distribuição de títulos demonstrou um padrão típico de área negativa no município de Cananéia e de baixa endemicidade nos demais. A investigação de casos revelou predominância de importados e, quando autóctones, sugestivos de transmissão oral. Foram observadas 1.261 unidades domiciliares com relato de presença de triatomíneos vetores (principais espécies: Panstrongylus megistus e Triatoma tibiamaculata), com um total de 5.338 amostras de sangue colhidas e 40 sororreagentes (0,75%). Exemplares adultos predominaram no intradomicílio e cerca da metade deles sem ingesta de sangue humano. Não foi observada diferença entre resultados de sorologia em moradores de casas com triatomíneos infectados e não infectados por Trypanosoma cruzi. CONCLUSÕES: Os resultados mostram ser necessário desenvolver um trabalho integrado entre instituições, que permita o isolamento e estudos de cepas e de isolados de tripanosomatídeos presentes em vetores, reservatórios (domésticos ou não) e humanos, com o objetivo de observar a interação dos pontos de vista evolutivo, de patogenia e de ecologia, de eventuais diferentes linhagens.

Epidemiología de la enfermedad de Chagas, Departamento General Paz, Argentina

OBJETIVO: El objetivo del presente trabajo fue investigar la colonización por triatominos en ambientes domésticos y peridomésticos y evaluar la seroprevalencia de infectados chagásicos en localidades rurales. MÉTODOS: La investigación se realizó en General Paz, Corrientes, Argentina. Las viviendas y peridomicilios se seleccionaron mediante un muestreo simple al azar. La búsqueda de triatominos se efectuó por el método captura/hora/hombre. Los insectos se identificaron taxonómicamente y se clasificaron según sexo y edad. La infección por Trypanosoma cruzi se detectó por observación directa de las heces al microscopio y por la técnica de la reacción en cadena de la polimerasa (PCR). Se evaluaron serológicamente a pobladores voluntarios mediante las técnicas de hemaglutinación indirecta (HAI) y ensayo inmunoenzimático (ELISA). Se hallaron los índices de infestación, densidad, colonización, infección natural y dispersión. RESULTADOS: Se analizaron 42 viviendas y 50 peridomicilios. En el domicilio los índices de infestación e infección de Triatoma infestans fueron 23,8 y 19,4 respectivamente. Los índices de densidad, colonización y dispersión fueron 2,1; 47,0 y 50,0 respectivamente. La infestación del peridomicilio por T. infestans fue 5,9% y por T. sordida 11,8%. T. sordida resultó infectada por Trypanosoma cruzi en un 2,0%. La seroprevalencia al T. cruzi de los 85 pacientes fue del 22,3%. CONCLUSIONES: Se estima que en el área investigada la transmisión del T. cruzi está interrumpida. Sin embargo, la presencia de infectados humanos y de T. infestans parasitadas posibilitan el reinicio del ciclo de transmisión. Se proponen acciones de rociado con insecticidas con el objeto de eliminar las poblaciones del vector.

Entomological aspects of Chagas' disease transmission in the domestic habitat, Argentina

OBJECTIVE: To study the risk of Trypanosoma cruzi domestic transmission using an entomological index and to explore its relationship with household's characteristics and cultural aspects. METHODS: There were studied 158 households in an endemic area in Argentina. Each household was classified according to an entomological risk indicator (number of risky bites/human). A questionnaire was administered to evaluate risk factors among householders. RESULTS: Infested households showed a wide range of risk values (0 to 5 risky bites/human) with skewed distribution, a high frequency of lower values and few very high risk households. Of all collected Triatoma infestans, 44% had had human blood meals whereas 27% had had dogs or chickens blood meals. Having dogs and birds sharing room with humans increased the risk values. Tidy clean households had contributed significantly to lower risk values as a result of low vector density. The infested households showed a 24.3% correlation between time after insecticide application and the number of vectors. But there was no correlation between the time after insecticide application and T. infestans' infectivity. The statistical analysis showed a high correlation between current values of the entomological risk indicator and Trypanosoma cruzi seroprevalence in children. CONCLUSIONS: The risk of T. cruzi domestic transmission assessed using an entomological index show a correlation with children seroprevalence for Chagas' disease and householders' habits.

Biogeografia, origem e distribuição da domiciliação de triatomíneos no Brasil

Considerando a distribuição atual da domicialiação triatomínea no Brasil, pode-se verificar sua associação com a referente aos espaços abertos. Estes podem ser considerados como naturais e artificiais. Os primeiros estão essencialmente situados nos domínios paisagísticos das caatingas, dos cerrados, totalmente incluídos em território brasileiro, e das pradarias mistas subtropicais pertencentes a paisagem que se estende além desses limites. Os outros são os originados da ação antrópica resultando em expansão da paisagem aberta principalmente em virtude da devastação da cobertura florestal do domínio tropical atlântico. Embora tentativamente, procurou-se aplicar o modelo de refúgios paleoecológicos e da existência de centros de endemismo às quatro espécies de domiciliação epidemiologicamente significante. O Triatoma sordida, Triatoma brasiliensis e Triatoma pseudomaculata parece ter seus centros de endemismo nos espaços abertos dos cerrados e das caatingas, enquanto o Panstrongylus megistus teria tido sua origem nas florestas do ambiente tropical atlântico. Quanto ao Triatoma infestans, sua área endêmica estaria localizada em território boliviano, de onde se dispersou e continua se dispersando pela ação do homem. A invasão domiciliar, ao que tudo indica, obedece a mecanismo oportunista propiciado por vários estímulos de abrigo e alimentação. Uma vez instalada, a domiciliação permite não apenas a sobrevivência, mas também a dispersão da espécie. Esses conceitos devem ser levados em conta nas campanhas de controle, uma vez que a probabilidade de sucesso aumenta com essa especialização do triatomíneo. A continuidade da ação antrópica sobre o ambiente, atualmente intensificada na região do domínio equatorial amazônico, resultará na expansão dos espaços abertos. Assim sendo, seja a custa de espécies locais seja a custa de espécies introduzidas, poderá ocorrer a domiciliação triatomínea, como problema de saúde pública, em região onde ainda não foi assinalada.

Infestación por triatominos en comunidades indígenas de Valledupar, Colombia

OBJETIVO: Calcular los índices infestación por triatominos en comunidades indígenas en Colombia. MÉTODOS: Se realizó estudio descriptivo en 19 comunidades indígenas del municipio de Valledupar Departamento de Cesar, Colombia. Durante junio a diciembre de 2007 se recolectaron triatominos por búsqueda activa en las viviendas de los indígenas. Los insectos luego fueron identificados por las claves de Lent & Wygodzinsky. Se desarrolló estudio del proceso infectivo en modelo animal y análisis enzimático de cepas de Trypanosoma cruzi, detectadas en heces de triatominos. RESULTADOS: Rhodnius prolixus presentó índice de densidad en las viviendas de 154,7%, Triatoma dimidiata de 102,45%, Triatoma maculata de 109,25% y Panstrogylus geniculatus de 0,3%. El índice promedio de infestación de las cuatro especies fue de 40,54% y, el de infección con T. cruzi de 9,4%. De cinco hemocultivos positivos para T. cruzi, tres se caracterizaron por isoenzimas, clasificándose en T. cruzi grupo I. El estudio de las biopsias reveló pocas características patológicas durante el proceso de infección con las cepas de T. cruzi aisladas de triatominos domiciliados. CONCLUSIÓN: Los altos índices de infestación por triatominos en las viviendas y el índice de infección por T. cruzi, evidencian la transmisión activa de la enfermedad de Chagas, situación que amerita la aplicación de medidas de control vectorial y el estudio seroepidemilógico de la población en riesgo. La identificación de las cepas de T. cruzi como grupo I concuerda con otros estudios realizados en esta región colombiana.

Trypanosoma cruzi strains from triatomine collected in Bahia and Rio Grande do Sul, Brazil

OBJECTIVE Collection of triatomines in domestic, peridomestic and sylvatic environments in states of Bahia and Rio Grande do Sul, Northeastern and Southern Brazil respectively, and isolation of Trypanosoma cruzi strains. METHODS First, the captured triatomines were identified using insect identification keys, then their intestinal content was examined by abdominal compression, and the samples containing trypanosomatid forms were inoculated in LIT medium and Swiss mice. RESULTS Six triatomine species were collected in cities in Bahia, namely Panstrongylus geniculatus (01), Triatoma melanocephala (11), T. lenti (94), T. pseudomaculata (02), T. sherlocki (26) and T. sordida (460), and two in cities in Rio Grande do Sul, namely T. circummaculata (11) and T. rubrovaria (115). Out of the specimens examined, T. cruzi was isolated from 28 triatomine divided into four different species: T. melanocephala (one), T. lenti (one), T. rubrovaria (16) and T. sordida (10). Their index of natural infection by T. cruzi was 6.4%. CONCLUSIONS The isolation of T. cruzi strains from triatomines found in domestic and peridomestic areas shows the potential risk of transmission of Chagas disease in the studied cities. The maintenance of those T. cruzi strains in laboratory is intended to promote studies that facilitate the understanding of the parasite-vector-host relationship.

Tentativas de criação de triatomíneos em laboratório por meio de alimentação "in vitro"

Para simplificar a criação de triatomíneos em laboratório, necessária, por exemplo, à execução do xenodiagnóstico e em estudos de caráter biológico, foi tentada alimentação "in vitro", mediante emprego de sangue, citratado ou desfibrinado, de galinhas abatidas em matadouro avícola. Para avaliação da eficácia desse propósito, observações de duas naturezas, com Triatoma infestans, tiveram lugar, através das seguintes medidas; a) análise do encadeamento de estádios ninfais sucessivos e da exeqüibilidade de satisfatórios acasalamentos, oviposição fértil e adequada reprodução; b) apreciação da infectividade e da viabilidade do Trypanosoma cruzi nos insetos. Como conclusão, ficou evidente que, a despeito da facilitação operacional, o intuito não conduziu ao êxito desejado, já que o procedimento clássico, usado como controle, mostrou-se sempre superior.

Possible oral transmission of acute Chagas' disease in Brazil

In October, 1986, 7 to 22 days after a meeting at a farm in Paraíba state, 26 individuals presented with a febrile illness associated with bilateral eyelid and lower limb edema, mild hepatosplenomegaly, lymphadenopathy and, occasionally a skin rash. A 11-year-old boy exhibited atrial premature complexes and a 74-year-old patient developed acute heart failure. In two patients hospitalized in São Paulo city, acute Chagas' disease was diagnosed by the demonstration of circulating Trypanosoma cruzi. At autopsy in a fatal case, acute Chagas' cardiomyopathy was demonstrated. Xenodiagnosis were positive in 9 out of 14 tested patients. A specific IgG immune response was found in all patients and specific IgM antibodies were identified in 20 out of 22 tested patients. A epidemiological survey showed the existence of Triatoma brasiliensis in the outbuildings of this farm, but none in the house where most of the guests stayed. A high rate of infection with Trypanosoma cruzi was found in opossums. These observations together with those related to the food consumed by the patients, lead the authors to suggest that the human infections resulted from oral contamination probably originating from naturally infected marsupials in the area or crushed infected bugs.

Lethality of triatomines (Hemiptera: Reduviiae), vectors of Chagas' disease, feeding on blood baits containing synthetic insecticides, under laboratory conditions

A laboratory study was conducted to test the toxicity of synthetic insecticides added to defibrinated sheep blood kept at room temperature and offered as food to the following triatomine species: Triatoma infestans, Panstrongylus megistus, Triatoma vitticeps, Triatoma pseudomaculata, Triatoma brasiliensis and Rhodnius prolixus. The insecticides used, at a concentration of 1g/l, were: HCH, DDT, Malathion and Trichlorfon, and the lethalithy observed at the end of a 7-day period varied according to the active principle of each. HCH was the most effective by the oral route, killing 100% of the insects, except P. megistus (95.7%) and T. pseudomaculata (94.1%). Trichlorfon killed the insects at rates ranging from 71.8% (T. vitticeps) to 98% (R. prolixus). Malathion was slightly less efficient, killing the insects at rates from 56.8% (T. vitticeps) to 97% (T.brasiliensis). DDT was the least effective, with a killing rate of 10% (T. vitticeps) to 75% (T.brasiliensis). Since the tests were performed at room temperature, we suggest that baits of this type should be tried for the control of triatomines in the field.

Hemolymphatic components in vectors of Trypanosoma cruzi: study in several species of the subfamily Triatominae (Hemiptera: Reduviidae)

The members of the subfamily Triatominae (Hemiptera : Reduviidae) comprise a great number of species of medical importance in the transmission of the T. cruzi (American trypanosomiasis). The aim of this study was to contribute to the knowledge about the chemical composition in proteins, lipids, lipoproteins, and carbohydrates of vectors of Chagas' disease corresponding to twelve members of the subfamily Triatominae. This study was carried out in ninphs of the fifth instar and adult males of the species: T. delpontei, T. dimidiata, T. guasayana, T. infestans, T. mazzotti, T. pallidipennis, T. patagonica, T. platensis, T. rubrovaria, T. sordida of the Triatoma genus, and D. maximus and P. megistus of the Dipatalogaster and Panstrongylus genera respectively. The results show on one hand, qualitative differences in the protein composition, and on the other hand, similarity in the lipoprotein profiles. Lipids, proteins, and carbohydrates did not show significant differences between species or/and stages.

Domestic vectors of Chagas' disease in three rural communities of Nicaragua

A triatomine survey was conducted in three rural settlements of Nicaragua (Santa Rosa, Quebrada Honda and Poneloya) where Chagas' disease is endemic, to determine rates of house infestation, evaluate the housing condition and to asess the performance of the María sensor box in detection of domestic vectors. A total of 184 households were selected and vectors were sought by the methods of timed manual capture and by sensor boxes. The sole vectors species found in this study was Triatoma dimidiata. Of the examined bugs 50, 60 and 33%, in the respective communities, were infected with T. cruzi. The rates of house infestation as determined by manual capture and sensor boxes were respectively, 48.3% and 54.2% in Santa Rosa, 29.8% and 51.2% in Quebrada Honda and in Poneloya 3.8 and 5.9% with significant difference between the methods in Quebrada Honda. When compared with the manual capture, the Maria sensor box detected vectors in 71.4% of positive houses in two of the communities but also was able to detect bugs in 39.3% and 41.1% of houses where manual capture had been negative. Housing condition was evaluated according to three structural parameters, in this way, in the first community 79.2% of houses were classified as bad, 20.8% as regular; in the second one 42.5% were bad and 57.5% regular, whereas in the third 62.5% of the houses were regular. Rates of infestation did not differ greatly between the different housing conditions. Our results show that the sensor box is as efficient as manual capture and could be implemented in our country.