Sôbre o Trypanosoma conorrhini, hemoparasito do rato transmitido pelo Triatoma rubrofasciata: presença do vector infectado na cidade do Rio de Janeiro

Os autores, após historiar a desccberta do Trypanosoma conorrhini (DONOVAN, 1909) (sinonimia Crithidia conorrhini DONOVAN, 1909, Trypanosoma boy lei LAFONT, 1912) no inseto transmissor e no hospedeiro vertebrado, o Rattus rattus diardi, fazem um estudo sumário do seu vector intermediário, o barbeiro cosmopolita Triatoma rubrofasciata (DE GEER, 1773) . Em seguida, referem a captura, no centro da cidade do Rio de Janeiro, Brasil, de um exemplar adulto dêste barbeiro parasitado por flagelados que foram identificados ao Trypanosoma conorrhini, fato êste pela primeira vez verificado no Novo Mundo. Certas formas evolutivas dêste parasito no inseto são muito semelhantes às do Schizotrypanum cruzi, mas os tripanosomas metacíclicos apresentam alguns caractéres morfológicos que permitem seu reconhecimento: os mais notáveis são o pequeno tamanho e a colocação do núcleo muito para trás, bem junto ao blefaroplasto. São dadas as medidas de 100 tripanosomas metaciclicos do T. conorrhini e de Schizotrypanum, cujo comprimento total médio foi, respectivamente, 13.88 u e 19.85 u. Nas seguintes espécies de barbeiro foi facilmente obtida a evolução do Trypanosoma conorrhini: Triatoma infestans, Triatoma vitticeps, Rhodnius prolixus e Panstrongylus megistus. Por inoculação de triatomas infectados foi obtida a transmissão do T. conorrhini ao camondongo branco, ao rato e ao macaco rhesus. As infecções foram muito fracas, custando-se a ver o tripanosoma no sangue, a fresco. Em gotas espêssas êle é encontrado com mais facilidade, mas o método de escôlha para verificação da sua presença no sangue dos animais é o xenodiagnóstico. Nesta prova o Triatoma infestans foi empregado com os resultados mais favoráveis. Por êste processo foi observada a infecção inaparente de um camondongo até 53 dias depois da inoculação. A forma sanguícola do T. conorrhini é um tripanosoma ttpico, de grande dimensões (40-60u), de extremidade posterior muito alongada e pontuda, membrana ondulante ampla e com largas pregas, blefaroplasto subcentral; em muitos indivíduos é notável uma estrutura particular, junto e à frente do blefaroplasto. Poucas horas depois de inoculados com as dejeções, os flagelados passam para a circulação, transformando-se em 3-4 dias nos grandes tripanosomas que acabam de ser descritos. O número de parasitos depende do número de flagelados inoculados. Não se conhece o processo de multiplicação do T. conorrhini no vertebrado, nunca tendo sido observados tripanosomas em via de divisão no sangue nem formas proliferativas nos tecidos. As sub-inoculações (de animal a animal) em geral não passam da primeira (MORISHITA, 1935). Tentativas até agora realizadas por outros pesquisadores no sentido de cultivar o T. conorrhini a partir do sangue de animais infectados, têm sido negativas. Por enquanto as pesquisas tendentes a determinar o hospedeiro vertebrado do parasito no Rio de Janeiro limitam-se a aplicação do xenodiagnós-tico em cinco ratos do Morro de Santo Antônio, cujo resultado...

Estudos anatômicos e histológicos sôbre a subfamília Triatominae (Hetroptera, Reduviidae): III parte: pesquisas sôbre o mecanismo da picada dos Triatominae

É analisado o mecanismo da picada dos Triatominae sob a consideração das observações até hoje conhecidas em Heteroptera e Homoptera. Leva à conclusão que os dados da literatura não chegam para esclarecer o mecanismo da picada dos Triatominae. Dos resultados das pesquisas e observações do animal vivo surge o seguinte quadro: Uma contração extremamente forte do músculo protrator pode mover as mandíbulas do adulto de Triatoma infestans para frente no máximo por 500 micra. Uma fonte de alimentação suficiente pode ser encontrada na epiderme do hospedeiro (homen) apenas numa profundidade de pelo menos 1000 a 1500 micra. O lábio nem pode ser encurtado nem dobrado. Correspondendo aos "fingerfoermigen Fortsaetzen" (WEBER) encontra-se na ponta do último segmento labial um mecanismo para a fixação das mandíbulas numa certa posição. Depois do esgotamento das possibilidades de contração dos protratores das mandíbulas as últimas são fixadas na posição atual. Em seguida segue uma dilatação dos protratores e depois do relaxamento do aparelho de fixação das mandíbulas os protratores são capazes de mover novamente as mandíbulas por quasi 500 micra para frente. Êsse processo pode ser repetido umas vêzes . Com o aparelho de fixação é ligado um mecanismo para a limpeza das mandíbulas na parte apical do último segmento labial em forma de uma escova. Na cabeça dos Triatominae é situado um órgão de contrôle muito complicado para estímulos mecânicos controlando a entrada das mandíbulas no tecido do hospedeiro. (O órgão receptivo correspondente para estímulos químicos é o órgão gustativo da epifaringe, já descrito umas vêzes.) O órgão representa um prolongamento do fim do tentório tubiforme dirigindo-se para frente. Perto da alavanca de articulação das mandíbulas o tubo, cheio de um líquido, aumenta-se formando uma vesícula que se insere na alavanca citada e na parede da cabeça diretamente ou por meio de tonofibrilas. Daqui o tubo continua pela antena inteira e insere-se, fechado no fim, por dois feixes de tonofibrilas fortes na extremidade do último segmento antenal. Imediatamente antes do órgão de Johnston insere-se no tubo do órgão de contrôle um scolopídio grande com a contrainserção na parede da antena (pedicelo). Cada tensão na vesícula na alavanca de articulação da mandíbula altera a pressão no líquido do órgão. A alteração da pressão é percebida pelo scolopídio. O scolopídio possui um aparelho terminal que representa uma invaginação da cutícula da parede antenal. Esta formação encontra-se também nos scolopídios do orgão de Johnston. O órgão pertence tanto às formas hematófagas dos gêneros Triatoma e Rhodnius como também à várias formas predatoras e fitófagas. Pode-se supor que o órgão é presente num grande número dos Heteroptera e que, eventualmente, representa uma parte do plano geral da ordem. O grau da evolução muito elevada deste órgão de controle indica que tem um papel importante para a manutenção da vida das espécies.

Miocardite no macaco Cebus após inoculações repetidas com Schizotrypanum cruzi

Transmission of Chagas disease is realized through contamination of ocular conjunctiva, mucosa or skin with infected dejections eliminated by the insect vectors of Schizotrypanum cruzi (Triatoma infestans, Panstrongylus megistus and Rhodnius prolixus). The triatomid bugs live in holes and craks in the walls, in beds, behind trunks, etc. Found in primitive mud huts covered with thatched roofs, and so the human dwellers have many chances to contract the disease, reinfections being reasonably more to expect than a single inoculation. Experimental work reproducing those natural conditions is welcomed as some important features in the pathologic picture of the disease such as the extensive myocardial fibrosis seen in chronic cases are still incompletely known. Microscopic changes were studied in the heart muscle of seven Cebus monkeys infected by S. cruzi. This animal survives the acute stage of the disease and so is particularly suited to experiments of long duration in which several inoculations of S. cruzi are performed. Three different strains of S. cruzi isolated from acute cases of Chagas' disease were employed. One monkey was injected in the skin with infected blood and necropsied after 252 days. Two monkeys were three times, and one, eight times infected in skin, one of them with contaminated blood, and two with contaminated blood and dejections from infected bugs. The necropsies were performed after 35, 95 and 149 days. One monkey was three times inoculated through the intact ocular conjunctiva (one time with infected blood, two times with dejections from infected bugs), and one time through the wounded buccal mucosa, and necropsied after 134 days. Another monkey was six times inoculated, four times through the intact ocular conjunctiva (one time with contaminated blood, three times with dejections from infected bugs) and two times injected in the skin with infected blood, and necropsied after 157 days. Finally, another monkey was nine times inoculated, four times through the intact ocular conjunctiva (one time with infected blood, and three times with dejections from infected bugs), and five times injected in the skin (four times with contaminated blood, and one time with dejections from infected bugs), and necropsied after 233 days. The microscopic picture was uniform presenting, however, considerable individual variations, and was represented by diffuse interstitial myocarditis, frequently more (marked in the right ventricle base of the heart), accompanied by lymphatic stasis. The infiltration consists of macrophages, plasma cells and lymphocytes, the cellular reaction having sometimes a perivascular distribution, involving the auriculo-ventricular system of conduction, endocardium, epicardium and cardiac sympathetic gangliae. The loss of cardiac muscle fibers was always minimal. Leishmanial forms of S. cruzi in myocardial fibers are scanty and, in two cases, absent. Fatty necrosis in the epicardium was noted in two cases. Obliterative changes of medium-sized branches of coronary arteries (hypersensitivity reaction?) and multiple infarcts of the myocardium was found in one instance. The diffuse myocarditis induced by S. cruzi in several species of monkeys of the genus Cebus observed after 233 days (several inoculations) and 252 days (single inoculation) is not associated with disseminated fibrosis such as is reported in chronic cases of Chagas' disease. Definite capacity of reversion is another characteristic of the interstitial myocarditis observed in the series of Cebus monkeys here studied. The impression was gained that repeated inoculation with S. cruzi may influence the myocardial changes differently according to the period between the reinoculations. A short period after the first inoculation is followed by more marked changes, while long periods are accompanied by slight changes, which suggests an active immunisation produced by the first inoculation. More data are required, however before a definite statement is made on this subject considering that individual variations, the natural capacity of reversion of the interstitial myocarditis and the employement of more than a species of Cebus monkeys probably exerts influence also in the results here reported.

Estudos anatômicos e histológicos sôbre a sub-família Triatominae (Heteroptera, Reduviidae): Parte XX: estudo comparado dos estigmas nas espécies mais importantes

É feito um estudo comparado sôbre a estrutura e o mecanismo dos estigmas de Triatoma vitticeps, Triatoma maculata, Triatoma sórdida, Triatoma brasiliensis, Rhodnius prolines e Panstrongyius megistus com os de Triatoma infestans, anteriormente já tratados. É, assim, elaborada uma tabela comparativa sôbre o tamanho e localização, dos espiráculos torácicos e abdominais, nas espécies citadas. Os estigmas mesotorácicos nas espécies do gênero Triatoma apresentam-se com várias modificações morfológicas, porém, não tão significantes como as encontradas em Panstrongyius megistus e Rhodnius prolixus. Seu mecanismo mostra acentuadas diferenças morfológicas. De um modo geral, os estigmas mesotorácicos possuem uma zona dorsal e outra ventral, que se diferenciam nas diferentes espécies e gêneros. A abertura do átrio para o exterior faz-se, perpendicularmente ao eixo maior do corpo do inseto. Nota-se uma região onde a cutícula é mais espêssa e que serve de base para a implantação dos músculos destinados ao seu funcionamento. O côndilo, que varia quanto à forma, nos diversos Triatomíneos, não é evidenciado nos estigmas metatorácicos. Os espiráculos metatorácicos encontram-se ocultos, totalmente, pelos escleritos do mesotórax e metatórax. O aspecto morfológico do mesmo é semelhante ao do mesotorácico, porém, apresenta algumas diferenças. Os espiraculos abdominais são em número de 8 pares. O 1.º par está situado na região látero-dorsal da membrana intersegmental, entre o tórax e o abdômen. Em Rhodnius prolixus o estigma apresenta-se circundado por zonas esclerosadas, em forma de listas. Nos demais Triatomíneos os escleritos estigmatíferos são semelhantes e menos reforçados. A forma e a estrutura e o mecanismo dêstes estigmas variam em todas as espécies citadas, Encontram-se 3 elementos de proteção à traquéia que se limita com o átrio. São eles: 1) rêde protetora que recobre parcialmente a entrada do estigma, semelhante a um peritrema; 2) côndilo interno que comanda a entrada e saída do ar das traquéias; e 3) falsos espinhos, situados no átrio e que variam de tamanho e espessura nos diferentes Triatomíneos. Finalmente, observa-se o VIII par de estigmas abdominais, cuja forma, mecanismo e localização o diferenciam dos demais. Êste par encontra-se citado por alguns autores, porém, na realidade, sua estrutura só é apresentada no presente trabalho.

Studies in search of a suitable experimental insect model for xenodiagnosis of hosts with Chagas' disease. 1 - Comparative xenodiagnosis with nine triatomine species of animals with acute infections by Trypanosoma cruzi

In search of a suitable vector species for xenodiagnosis of humans and animals with chronic Chagas' disease we first investigated the reactions of different vector species to acute infection with Trypanosoma cruzi. Vector species utilized in this study were: Triatoma infestans, Rhodnius prolixus and Triatoma dimidiata, all well adapted to human habitats; Triatoma rubrovaria and Rhodnius neglectus both considered totally wild species; Panstrongylus megistus, Triatoma sordida, Triatoma pseudomaculata and Triatoma brasiliensis, all essentially sylvatic but some with domiciliary tendencies and others restricted to peridomestic biotopes with incipient colonization of human houses after successful eradication of T. infestans. Results summarized in Table IV suggest the following order of infectivity among the 9 studied vector species: P. megistus with 97.8% of infected bugs, T. rubrovaria with 95% of positive bugs a close second followed by T. Pseudomaculata with 94.3% and R. neglectus with 93.8% of infected bugs, almost identical thirds. R. prolixus, T. infestans and T. dimidiata exhibited low infection rates of 53.1%, 51.6% and 38.2% respectively, coupled with sharp decreases occuring with aging of infection (Fig. 1). The situation was intermediate in T. brasiliensis and T. sordida infection rates being 76.9% and 80% respectively. Results also point to the existence of a close correlation between prevalence and intensity of infection in that, species with high infection rates ranging from 93.8% to 97.8% exhibited relatively large proportions of insects (27.3% - 33.5%) harbouring very dense populations of T. cruzi. In species with low infection rates ranging from 38.2% to 53.1% the proportion of bugs demonstrating comparable parasite densities was at most 6%. No differences attributable to blood-meal size or to greater susceptibility of indigenous vector species to parasites of their own geographical area, as suggested in earlier...

Studies on Trypanosoma rangeli Tejera 1920. VII - Its effect on the survival of infected triatomine bugs

The pathological effects of Trypanosoma rangeli on Rhodnius prolixus and R. robustus, and the relation of mortality to infection, were studied under laboratory conditions. Frequent observations revealed that when the first instar nymphs of R. prolixus and R. robustus were infected with T. rangeli, survival of the bugs during the stages of development to the adult stage decreased. This decrease was statistically significant when compared with uninfected control-bugs, indicating that T. rangeli is pathogenic for both species of triatomine. In R. prolixus the most affected nymphal stages were the first, second and fifth instars, where a higher mortality was also observed. In R. robustus a progressive increase of the mortality from the first to fifth instars, was observed. The pathogenicity of T. rangeli as measured by overall mortality was the same in R. prolixus and R. robustus. The possible pathogenic mechanism of T. rangeli in triatomine-bugs and its epidemiological implications, are discussed.

Estudo anatômico e morfométrico dos folículos testiculares de algumas espécies de Triatominae (Hemiptera: Reduviidae)

Os folículos testiculares nas ordens dos insetos, vêm demonstrando papel relevante sob o ponto de vista da Sistemática e Filogenia. Nos triatomíneos, os folículos testiculares são em número de sete, sendo que o comprimento e a espesura variam consideravelmente entre os gêneros, e pouco entre as espécies. Verificou-se que em Rhodnius spp existem dois folículos longos e grossos e cinco curtos e fuinnos; em Triatoma spp existem dois longos e finos, dois médios e grossos e três curtos e finos; e em Panstrongylus spp existem sete aproximadamente iguais. Através das medições do comprimento dessas estruturas, evidenciou-se que os valores das médias e desvios padrão das médias, foram muito próximos entre as espécies do mesmo gênero, tornando-se inviável estabelecer limites que pudessem caracterizá-las. Em busca de resultados mais precisos, fizeram-se comparações interespecíficas através da análise de variância: entre as diferentes espécies do mesmo gênro (comparação simultânea); entre cada duas espécies do mesmo gênro (comparação aos pares); e entre espécies iguais de procedência diferentes. Os resultados obtidos não demostraram possibilidades de utilização na Sistemática, contudo foram promissores no que concerne à Filogenia, desde que seja observado um número maior de espécies.

The Azadirachtins: potent insect growth inhibitors

In the course of their coevolution with insects, plants have learnt to protect themselves by chemical means. Semiochemical act as antifeedants or deterrents, others by disrupting growth and development. By use of the Epilachna varivestis bioassay we isolated from Azadirachta indica seed a group of triterpenoids which interfee with larval growth and development in ppm range. Main components are the azadirachtins A and B with identical biological activity. Various other azadirachtins were obtained, either as minor seed components or by chemical modification of the naturally occuring compounds. Structure vs. activity relation studies enabled us to postulate a basic structural element that should still be biologically active and with much simpler chemical structure than natural compounds. What underlies the biological activity of these insect growth inhibitors? Their interference with the hormonal regulation of development and reproduction has been studied in Locusta migratoria and Rhodnius prolixus. In addition, tritiated dihydroazadirachtin A was used. With this approach, a precise correlation between administered dose, resulting effects, and retention of the compound was established. The azadirachtins either interrupt, delay, or deviate whole developmental programs. Results from these studies provide another chemical probe for studies in insect endocrinology and physiology.

Estudo comparado do ciclo evolutivo de triatomíneos a duas temperaturas

We studied the life cicle of several triatominae species: Dipetalogaster maximus (Uhler, 1894); Panstrongylus herreri Wygodzinsky, 1948; Panstrongylus megistus (Burmeister, 1835); Rhodnius ecuadoriensis Lent & Leon, 1958; Rhodnius nasutus Stal, 1859; Rhodnius neglectus Lent, 1954; Rhodnius pictipes Stal, 1872; Rhodnius prolixus Stal, 1859; Rhodnius robustus Larrousse, 1927; Triatoma brasiliensis Neiva, 1911; Triatoma infestans (Klug, 1834); Triatoma maculata (Erichson, 1848); Triatoma matogrossensis Leite & Barbosa, 1953; Triatoma platensis Neiva, 1913; Triatoma protracta (Uhler, 1894); Triatoma sordida (Stal, 1859); Triatoma tibiamaculata (Pinto, 1926) e Triatoma vitticeps (Stal, 1859) (Hemiptera, Reduviidae). The main purpose of the study was to obtain information to improve control measures specially in those peridomiciliar species. The experiments were performed in two climatized chambers, both with an humidity of 70 ± 5% and photoperiod of 12 hours. One was maintained at 25 ± 0,5ºC and the other at 30 ± 1ºC.

Aspects of classification of Hemiptera hemocytes from six triatomine species

The objective of this work was to characterize, and compare different morphological types of hemocytes of Rhodnius prolixus, Rhodnius, Rhodnius neglectus, Triatoma infestans, Panstrongylus megistus, and Dipetalogaster maximus. This information provides the basis for studying the cellular immune systems of these insects. Seven morphological hemocyte types wereidentified by phase-contrast microscopy: prohemocytes, plasmatocytes, granular cells, cytocytes, oenocytoids, adipohemocytes and giant cells. All seven types of hemocytes are not present in every species. For example, adipohemocytes and oenocytoids were not observed in P. megistus and P. infestans, and giant cells were rarely found in any of the species studied. The hemocytes of rhodnius and Dipetalogaster are more similar to each other than those from Triatoma and Panstrongylus which in turn closely resemble each other. Emphasis is placed on methodological problems arising in this work wicah are discussed in detail.

The importance of the opossum (Didelphis albiventris) as a reservoir for Trypanosoma cruzi in Bambuí, Minas Gerais state

In a survey realized on the sylvatic and peridomestic environment at Bambuí county, Minas Gerais State, 44 (37.9%) out of 116 opossums (Didelphis albiventris) captured were found to be naturally infected with Trypanosoma cruzi. One handred and forty three parasite samples were obtanied from 43 infected opossums using simultaneously hemoculture, xenodiagnosis (Triatoma infestans, Panstrongylus megistus and Rhodnius neglectus) and examination of anal glands contents. The parasite samples were characterized according to six isoenzyme patterns. All samples, independently of the method of isolation, presented an isoenzyme pattern similar to the standard T. cruzi Z1, showing that either xenodiagnosis or hemoculture can used without selecting parasite subpopulation from naturally infected opossums. Preveous isoenzyme patterns reported for human T.cruzi isolates from same region were completely different. This isoenzyme dissimilarity between sylvatic and domiciliar environments suggests the existence of two independent T. cruzi transmission cycles in Bambuí. The epidemiological implicatinos of these results are discussed.

Sexual distinction between 5th instar nymphs of six species of Triatominae (Hemiptera, Reduviidae)

The aim of this paper is to present the analysis of sexual morphological differences observed in 5th instar nymphs of the following species: Panstrongylus megistus; Rhodnius neglectus; Triatoma brasiliensis; T. infestans; T. matogrosensis and T. tibiamaculata. Male and female nymphs were examined and photographed with a Scanning Electron Microscope. The 9th segment dimensions of dorsal and ventral faces were determined through a Profile Projector. Results and statistical analysis showed significant differences: the 9th sternite is significantly broader in male than female nymphs, while in five species; tergites in female nymphs are broad and in male are narrow.

Cytogenetics as a tool for Triatomine species distinction (Hemiptera-Reduviidae)

Several cytogenetic traits were tested a species diagnostic characters on five triatomine species: Rhodnius pictipes, R. nasutus, R. robustus, Triatoma matogrossensis and T. pseudomaculata. Four of them are described for the first time. The detailed analysis of the meiotic process and the application of C-banding allowed us to identify seven cytogenetic characters wich result useful to characterize and differentiate triatomine species.

Relative susceptibility of different stages of Rhodinius Prolixus to the entomopathogenic Hyphomycete Beauveria Bassiana

Laboratory bioassays were conducted to determine the relative suscepbility of eggs, 1st-, 3rd-, 5th- instar nymphs and adults of Rhodnius prolixus to one isolate of the entomopathogenic hyphomycete, Beauveria bassiana. Treatments consisted of directly spraying on insects of increasing doses of inoculum (3 x 10* to 3 x 10 (elevated to 5th potency) conidia per cm*). Mortality due to all doses of conidia was very high in the five tested stages of the target insect. Experiments on eggs demonstrated that the fungal isolate was able to kill eggs before they hatched. Both time-mortality and dose-mortality responses showed that the susceptibility of R. prolixus varied according to its stage of development and increased with age. As matter of fact, at the dose of 3 x 10* conidia per cm*, LD50 varied between 11.2 days in 1st-instar nymphs and 6.4 days in both 5th-instar nymphs and adults. Comparison of LD50 permitted to estimate that 1st-instar nymphs were about 700-fold less susceptible than the two oldest stages

Aggregation pheromone in five species of Triatominae (Hemiptera: Reduviidae)

An aggregation pheromone found in the faeces of 5th instar nymphs and adults of Triatoma mazzottii Usinger, Triatoma longipennis Usinger, Triatoma pallidipennis (Stal), Triatoma barberi Usinger and Rhodnius prolixus (Stal) was studied under laboratory conditions. Bioassays were performed using a 30 cm-diameter arena and wind tunnel. T. longipennis nymphs showed a stronger response than the other triatomine nymphs tested. There were no significant differences in faecal attractiveness to nymphs, but the faeces of T. longipennis and T. pallidipennis were most active. The responses of all species to male and female faeces of T. mazzottii was significantly different, but there was no significant difference in the responses of the development stages to male and female faeces of T. mazzottii. However, male faeces were more active than female faeces. The feeding status of nymphs did not affect the response.