Experimental Hybrids of the Triatoma brasiliensis Species Complex Show Higher Susceptibility to the Trypanosoma cruzi Infection Than Their Parentals.

The Triatoma brasiliensis species complex is a monophyletic group encompassing two subspecies and six species. Recently, a hybrid zone of members of this complex was recorded in the state of Pernambuco. Questions concerning the capability of the hybrids to become infected with Trypanosoma cruzi have been raised. This study aimed to compare the susceptibility of Triatoma b. brasiliensis, Triatoma juazeirensis, and their experimental hybrids to infection with T. cruzi. We infected the parentals and their experimental hybrids (obtained through reciprocal crosses) through artificial feeding with citrated rabbit blood, to which the TcI 0354 strain of T. cruzi had been added. The insects were weighed before and after feeding on the rabbit blood, and then they were dissected on the 10th, 20th, and 30th day after infection. Both the hybrids and the parentals remained infected throughout the experiment. The parasite was mostly found in the epimastigote form. The number of epimastigotes was significantly lower in the stomach and small intestine of T. juazeirensis than in the hybrids or in T. b. brasiliensis. A significantly higher percentage of metacyclic trypomastigotes was detected in the small intestine and rectum of the hybrids. Hybrids demonstrated higher susceptibility to the TcI 0354 strain than their parentals, opening up new avenues to be investigated.

Oral Trypanosoma cruzi Acute Infection in Mice Targets Primary Lymphoid Organs and Triggers Extramedullary Hematopoiesis.

During the acute phase of Chagas disease, Trypanosoma cruzi circulation through the bloodstream leads to high tissue parasitism in the host. In primary lymphoid organs, progenitor cell reduction paralleled transient immunosuppression. Herein we showed that acute oral infection in mice promotes diffuse parasitism in bone marrow cells at 14 and 21 days post-infection (dpi), with perivascular regions, intravascular regions, and regions near the bone being target sites of parasite replication. Phenotypic analysis of hematopoietic differentiation in the bone marrow of infected mice showed that the cell number in the tissue is decreased (lineage-negative and lineage-positive cells). Interestingly, analysis of hematopoietic branching points showed that hematopoietic stem and progenitor cells (HSPCs) were significantly increased at 14 dpi. In addition, the pool of progenitors with stem plasticity (HSC-MPP3), as well as multipotent progenitors (MPPs) such as MPP4, also showed this pattern of increase. In contrast, subsequent progenitors that arise from MPPs, such as common lymphoid progenitors (CLPs), lymphoid-primed MPPs (LMPPs), and myeloid progenitors, were not enhanced; conversely, all presented numeric decline. Annexin V staining revealed that cell death increase in the initial hematopoietic branching point probably is not linked to CLPs and that myeloid progenitors decreased at 14 and 21 dpi. In parallel, our investigation provided clues that myeloid progenitor decrease could be associated with an atypical expression of Sca-1 in this population leading to a remarkable increase on LSK-like cells at 14 dpi within the HSPC compartment. Finally, these results led us to investigate HSPC presence in the spleen as a phenomenon triggered during emergency hematopoiesis due to mobilization or expansion of these cells in extramedullary sites. Splenocyte analysis showed a progressive increase in HSPCs between 14 and 21 dpi. Altogether, our study shows that the bone marrow is a target tissue in T. cruzi orally infected mice, leading to a hematopoietic disturbance with LSK-like cell bias accounting on HSPCs possibly affecting myeloid progenitor numbers. The LMPP and CLP reduction converges with defective thymocyte development. Lastly, it is tempting to speculate that the extramedullary hematopoiesis seen in the spleen is a mechanism involved in the hematological maintenance reported during the acute phase of oral T. cruzi infection.

Proteome of the Triatomine Digestive Tract: From Catalytic to Immune Pathways; Focusing on Annexin Expression.

Rhodnius prolixus, Panstrongylus megistus, Triatoma infestans, and Dipetalogaster maxima are all triatomines and potential vectors of the protozoan Trypanosoma cruzi responsible for human Chagas' disease. Considering that the T. cruzi's cycle occurs inside the triatomine digestive tract (TDT), the analysis of the TDT protein profile is an essential step to understand TDT physiology during T. cruzi infection. To characterize the protein profile of TDT of D. maxima, P. megistus, R. prolixus, and T. infestans, a shotgun liquid chromatography-tandem mass spectrometry (LC-MS/MS) approach was applied in this report. Most proteins were found to be closely related to metabolic pathways such as gluconeogenesis/glycolysis, citrate cycle, fatty acid metabolism, oxidative phosphorylation, but also to the immune system. We annotated this new proteome contribution gathering it with those previously published in accordance with Gene Ontology and KEGG. Enzymes were classified in terms of class, acceptor, and function, while the proteins from the immune system were annotated by reference to the pathways of humoral response, cell cycle regulation, Toll, IMD, JNK, Jak-STAT, and MAPK, as available from the Insect Innate Immunity Database (IIID). These pathways were further subclassified in recognition, signaling, response, coagulation, melanization and none. Finally, phylogenetic affinities and gene expression of annexins were investigated for understanding their role in the protection and homeostasis of intestinal epithelial cells against the inflammation.

Vector competence and feeding-excretion behavior of Triatoma rubrovaria (Blanchard, 1843) (Hemiptera: Reduviidae) infected with Trypanosoma cruzi TcVI.

BACKGROUND: Several studies addressed changes on the insect vector behavior due to parasite infection, but little is known for triatomine bugs, vectors of Trypanosoma cruzi, the etiological agent of Chagas disease. We assessed infection rates and metacyclogenesis of T. cruzi (TcVI) in fifth-instar nymphs of Triatoma rubrovaria comparing with the primary vector Triatoma infestans. Also, biological parameters related to feeding-excretion behavior were evaluated aiming to identify which variables are most influenced by T. cruzi infection. METHODOLOGY/PRINCIPAL FINDINGS: Fifth-instar nymphs of T. rubrovaria and T. infestans were fed on mice infected with T. cruzi (TcVI). We compared the presence and the number of parasite evolutive forms in excreta of both triatomine species at 30, 60 and 90 days post-infection (dpi) with traditional statistical analyses. Moreover, both species were analyzed through generalized linear models and multinomial logistic regression hypotheses for seven behavioral parameters related to host-seeking and feeding-excretion. Triatoma rubrovaria and T. infestans had similar overall infection and metacyclogenesis rates of T. cruzi TcVI in laboratory conditions. Regarding vector behavior, we confirmed that the triatomine's tendency is to move away from the bite region after a blood meal, probably to avoid being noticed by the vertebrate host. Interspecific differences were observed on the volume of blood ingested and on the proportion of individuals that excreted after the blood meal, revealing the higher feeding efficiency and dejection rates of T. infestans. The amount of ingested blood and the bite behavior of T. rubrovaria seems to be influenced by TcVI infection. Infected specimens tended to ingest ~25% more blood and to bite more the head of the host. Noteworthy, in two occasions, kleptohematophagy and coprophagy behaviors were also observed in T. rubrovaria. CONCLUSIONS/SIGNIFICANCE: Laboratory infections revealed similar rate of T. cruzi TcVI trypomatigotes in excreta of T. rubrovaria and T. infestans, one of the most epidemiological important vectors of T. cruzi. Therefore, TcVI DTU was able to complete its life cycle in T. rubrovaria under laboratory conditions, and this infection changed the feeding behavior of T. rubrovaria. Considering these results, T. rubrovaria must be kept under constant entomological surveillance in Rio Grande do Sul, Brazil.

Oral Route Driven Acute Trypanosoma cruzi Infection Unravels an IL-6 Dependent Hemostatic Derangement.

Oral transmission of Trypanosoma cruzi, the etiologic agent of Chagas disease, is presently the most important route of infection in Brazilian Amazon. Other South American countries have also reported outbreaks of acute Chagas disease associated with food consumption. A conspicuous feature of this route of transmission is presenting symptoms such as facial and lower limbs edema, in some cases bleeding manifestations and risk of thromboembolism are evident. Notwithstanding, studies that address this route of infection are largely lacking regarding its pathogenesis and, more specifically, the crosstalk between immune and hemostatic systems. Here, BALB/c mice were orally infected with metacyclic trypomastigotes of T. cruzi Tulahuén strain and used to evaluate the cytokine response, primary and secondary hemostasis during acute T. cruzi infection. When compared with control uninfected animals, orally infected mice presented higher pro-inflammatory cytokine (TNF-α, IFN-γ, and IL-6) serum levels. The highest concentrations were obtained concomitantly to the increase of parasitemia, between 14 and 28 days post-infection (dpi). Blood counts in the oral infected group revealed concomitant leukocytosis and thrombocytopenia, the latter resulting in increased bleeding at 21 dpi. Hematological changes paralleled with prolonged activated partial thromboplastin time, Factor VIII consumption and increased D-dimer levels, suggest that oral T. cruzi infection relies on disseminated intravascular coagulation. Remarkably, blockade of the IL-6 receptor blunted hematological abnormalities, revealing a critical role of IL-6 in the course of oral infection. These results unravel that acute T. cruzi oral infection results in significant alterations in the hemostatic system and indicates the relevance of the crosstalk between inflammation and hemostasis in this parasitic disease.

Asymptomatic Plasmodium infection in a residual malaria transmission area in the Atlantic Forest region: Implications for elimination.

INTRODUCTION: Elimination of malaria in areas of interrupted transmission warrants careful case assessment to avoid the reintroduction of this disease. Occasional malaria cases are reported among visitors of the Atlantic Forest area of Brazil, while data on residents of this area are scarce. METHODS: A sectional study was carried out to examine 324 individuals living in a municipality where autochthonous cases were detected. RESULTS: Asymptomatic Plasmodium infections were detected in 2.8% of the individuals by polymerase chain reaction (PCR), with one case of P. falciparum (0.3%), two cases of P. vivax (0.6%), and six cases of P. malariae (1.9%). The thick blood smears were negative in all individuals. Serological tests performed in 314 subjects were reactive in 11.1%, with 3.5% for P. falciparum and 7.7% for P. vivax. A subsample of 42 reactive individuals for any Plasmodium species showed P. malariae in 30.9% of specimens. Individuals who entered the Atlantic Forest region were 2.7 times more likely to exhibit reactive serology for P. vivax compared with individuals who did not enter this region (p<0.05). Children <15 years had a higher chance of reactive serology for P. falciparum and P. vivax than individuals ≥15 years of age (p<0.05). Individuals living in the Paraiso district had a higher chance of reactive serology for P. vivax compared to other districts (p<0.05). No associations were found between sex, past exposure to malaria, or serological response to antibodies of any Plasmodium species. CONCLUSIONS: The implications of these results for the elimination of malaria were discussed.

Asymptomatic plasmodium infection in a residual malaria transmission area in the atlantic forest region: implications for elimination

Abstract INTRODUCTION: Elimination of malaria in areas of interrupted transmission warrants careful case assessment to avoid the reintroduction of this disease. Occasional malaria cases are reported among visitors of the Atlantic Forest area of Brazil, while data on residents of this area are scarce. METHODS: A sectional study was carried out to examine 324 individuals living in a municipality where autochthonous cases were detected. RESULTS: Asymptomatic Plasmodium infections were detected in 2.8% of the individuals by polymerase chain reaction (PCR), with one case of P. falciparum (0.3%), two cases of P. vivax (0.6%), and six cases of P. malariae (1.9%). The thick blood smears were negative in all individuals. Serological tests performed in 314 subjects were reactive in 11.1%, with 3.5% for P. falciparum and 7.7% for P. vivax. A subsample of 42 reactive individuals for any Plasmodium species showed P. malariae in 30.9% of specimens. Individuals who entered the Atlantic Forest region were 2.7 times more likely to exhibit reactive serology for P. vivax compared with individuals who did not enter this region (p<0.05). Children <15 years had a higher chance of reactive serology for P. falciparum and P. vivax than individuals ≥15 years of age (p<0.05). Individuals living in the Paraiso district had a higher chance of reactive serology for P. vivax compared to other districts (p<0.05). No associations were found between sex, past exposure to malaria, or serological response to antibodies of any Plasmodium species. CONCLUSIONS: The implications of these results for the elimination of malaria were discussed.

Lipoproteins from vertebrate host blood plasma are involved in Trypanosoma cruzi epimastigote agglutination and participate in interaction with the vector insect, Rhodnius prolixus.

Chagas disease, infecting ca. 8 million people in Central and South America, is mediated by the protozoan parasite, Trypanosoma cruzi. The parasite is transmitted by the bite of blood sucking triatomine insects, such as Rhodnius prolixus, that had previously fed on parasite-infected vertebrate blood and voided their contaminated feces and urine into the wound. The stages of the parasite life cycle in both the insect vector and human host are well-known, but determinants of infection in the insect gut are complex and enigmatic. This paper examines the possible role of the R. prolixus gut agglutinins in the parasite life cycle. The results, derived from gut extracts made from R. prolixus fed on various diets with different vertebrate blood components, and cross adsorption experiments, showed for the first time that R. prolixus has two distinct gut agglutinins originating from their vertebrate blood meal, one for T. cruzi (the parasite agglutinin, PA) and the other for the erythrocytes (the hemagglutinin, HA). Again, uniquely, the results also demonstrate that these two agglutinins are derived, respectively, from the plasma and erythrocyte components of the vertebrate blood. Subsequent experiments, examining in more detail the nature of the plasma components forming the T. cruzi PA, used fractionated extracts of the vertebrate plasma (high density lipoprotein, HDL; low density lipoprotein, LDL, and delipidated plasma) in agglutination assays. The results confirmed the identity of the PA as a high density lipoprotein (HDL) in the plasma of the vertebrate blood meal which agglutinates parasites in the R. prolixus gut. In addition, the use of single or double labeled HDL in fluorescence and confocal microscopy showed the interaction of the labeled HDL with the parasite surface and its internalization at later times. Finally, results of T. cruzi parasitization of R. prolixus, incorporating various vertebrate blood components, resulted in highly significant increases in infectivity in the presence of HDL from the 2nd day of infection, thus confirming the important role of this molecule in T. cruzi infection of R. prolixus.

Aspectos da biologia de diferentes espécies de triatomíneos sobre o desenvolvimento de tripanossomatídeoscaracterização de serpina, aglutinina e microbiota intestinal / Aspects of the biology of different species of insects on the development of the serpin tripanossomatídeoscaracterização, agglutinin and intestinal microbiota

Na família Reduviidae, algumas espécies da subfamília Triatominae são vetores de Trypanosoma cruzi e Trypanosoma rangeli. Este último causa apenas uma infecção transitória nos vertebrados e desenvolve a parte do seu ciclo nos triatomíneos, invadindo a hemocele e atingindo posteriormente as glândulas salivares. Já o T. cruzi é o agente causador da doença de Chagas e desenvolve o seu ciclo no hospedeiro invertebrado, ao longo do tubo digestivo com as formas infectivas para o mamífero sendo liberadas nas fezes do inseto durante a alimentação sanguínea subsequente à infectiva. Tanto o sobrenadante da hemolinfa quanto o extrato do tubo digestivo dos triatomíneos apresentam atividades de hemaglutinação (HA) contra eritrócitos de coelho e de aglutinação de parasitos (PA), contra formas de cultura de T. cruzi. Esta atividade foi encontrada em todas as espécies testadas, Rhodnius prolixus, Rhodnius neglectus, Rhodnius brethesi, Triatoma infestans, Panstrongylus megistus e Dipetalogaster maximus. A partir de testes de variação da dieta, de inibição e de adsorção foi demonstrado que as atividades de PA e de HA do extrato de tubo digestivo de R. prolixus são decorrentes de moléculas distintas. PA está associada ao HDL presente no plasma alimentar e sua atividade foi reduzida por alguns inibidores de interação hidrofóbica...

Além disso, verificou-se a redução da infecção de R. prolixus por T. cruzi em insetos alimentados apenas com eritrócitos ou com plasma delipidado no lugar do plasma normal. Na hemolinfa, tanto a atividade de HA quanto a de PA são maiores em P. megistus em comparação com as outras espécies de triatomíneos. Paralelamente, verificou-se que o perfil de proteínas de hemolinfa dessa espécie (SDS-PAGE), mostra uma banda majoritária, de 38,8KDa, cuja correspondência foi encontrada também em P. lutzi, mas não nos outros gêneros estudados. A análise da banda submetida à digestão tríptica e espectrometria de massa indicou que parte dos peptídeos apresentava sequências homólogas com serpinas descritas em outros organismos. A partir da informação da sequência de aminoácidos desses peptídeos foram desenhados primers degenerados para identificar o gene da serpina em uma biblioteca de cDNA obtida do corpo gorduroso de P. megistus. Por essa abordagem foi obtida a sequência gênica codificante e deduzida à sequência de aminoácidos da serpina (PMSRP1)Posteriormente, foi definida a estrutura espacial por modelagem mostrando que PMSRP1 é similar às serpinas caracterizadas em outros insetos. Resultados preliminares sugerem que PMSRP1 está inativa na hemolinfa, pelo rompimento da ligação Arg-Ile localizada na alça do centro reativo (RCL)...

O gene de PMSRP1 é expresso em diferentes tecidos de P. megistus, em intensidade decrescente para hemócitos, corpo gorduroso, glândula salivar, intestino e estômago. Finalmente, também foi identificada, pela técnica de DGGE, a estrutura da microbiota intestinal de diferentes espécies de triatomíneos, sendo relacionado para cada gênero um padrão de espécies de bactérias da microbiota, incluindo populações de diferentes gêneros como Serratia, Arsenophonus, Rohrkolberia ou Wolbachia. Comparativamente, a riqueza relativa da microbiota intestinal dos triatomíneos analisados apresentou um grau de complexidade decrescente para as espécies Triatoma infestans, Rhodnius prolixus, Panstrongylus megistus e Dipetalogaster maximus...

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.

Morphobiological aspects of Rhodnius brethesi Matta, 1919 ( Hemiptera:Reduviidae ) from the Upper and Middle Negro River, Amazon region of Brazil: I - scanning electron microscopy

The occurrence of autochthonous cases of Chagas disease in the Amazon region of Brazil over recent decades has motivated an intensification of studies in this area. Different species of triatomines have been identified, and ten of these have be proven to be carriers of the parasite Trypanosoma cruzi or " cruzi-like " parasites. Studies conducted in the municipalities of Santa Isabel do Rio Negro and Barcelos, located on the Upper and Middle of the Negro River, microregion of Negro River, state of Amazonas have confirmed not only that Rhodnius brethesi is present in the palm tree Leopoldinia piassaba, but also that this insect was recognized by palm fiber collectors. A morphological study of eyes, inter-ocular and inter-ocellar regions, antennae, buccula, labrum, rostrum, stridulatory sulcus and feet, including the apex of the tibia, spongy fossette and ctenidium was conducted by scanning electron microscopy. The buccula and the stridulatory sulcus presented notable differences in specimens of different genera and also of different species. These data make it possible to suggest that the details presented in these structures can be included as diagnostic characteristics to be used in new dichotomous keys, thereby contributing towards studies of taxonomy and systematics and furnishing backing for comparative analysis of specimens collected from different localities.

Morphobiological aspects of Rhodnius brethesi Matta, 1919 (Hemiptera:Reduviidae) from the Upper and Middle Negro River, Amazon region of Brazil: I - scanning electron microscopy.

The occurrence of autochthonous cases of Chagas disease in the Amazon region of Brazil over recent decades has motivated an intensification of studies in this area. Different species of triatomines have been identified, and ten of these have be proven to be carriers of the parasite Trypanosoma cruzi or " cruzi-like " parasites. Studies conducted in the municipalities of Santa Isabel do Rio Negro and Barcelos, located on the Upper and Middle of the Negro River, microregion of Negro River, state of Amazonas have confirmed not only that Rhodnius brethesi is present in the palm tree Leopoldinia piassaba, but also that this insect was recognized by palm fiber collectors. A morphological study of eyes, inter-ocular and inter-ocellar regions, antennae, buccula, labrum, rostrum, stridulatory sulcus and feet, including the apex of the tibia, spongy fossette and ctenidium was conducted by scanning electron microscopy. The buccula and the stridulatory sulcus presented notable differences in specimens of different genera and also of different species. These data make it possible to suggest that the details presented in these structures can be included as diagnostic characteristics to be used in new dichotomous keys, thereby contributing towards studies of taxonomy and systematics and furnishing backing for comparative analysis of specimens collected from different localities.

Dynamics of evolution and resistance to starvation of Triatoma vitticeps (Stal 1859) (Reduviidae: Triatominae), submitted to two different regimens of food deprivation.

The resistance to starvation of Triatoma vitticeps has been analyzed comparatively, according to different regimens of food deprivation under laboratory conditions. One cohort, composed of 100 specimens of each evolutionary nymphal stage, was submitted to continuous fasting until death; the second group, arranged in the same way, was fed once on chicken. Through this work, it was possible not only to compare the results obtained from the first group with other authors' results but, also, to analyze the dynamics of evolution, molting, longevity and the increase in insect longevity, in the second group. The average values recorded for survival time revealed statistical differences between the two groups. Among the important results detected, there is one that deserves to be emphasized: the incredible increase in longevity among insects that received only one feed - an average increase in survival time that reached 2.95 to 3.30 times in nymphs of 3rd and 4th stages, respectively. One 5th stage nymph survived for up to 350 days and the females may triplicate their survival rate, what represents an important epidemiological factor. The knowledge about this type of biological characteristic of T. vitticeps may contribute to prevent the domiciliation of this species, what seems to be incipient in some municipal districts, in Brazil.

Dynamics of evolution and resistance to starvation of Triatoma vitticeps (Stal 1859) (Reduviidae: Triatominae), submitted to two different regimens of food deprivation

The resistance to starvation of Triatoma vitticeps has been analyzed comparatively, according to different regimens of food deprivation under laboratory conditions. One cohort, composed of 100 specimens of each evolutionary nymphal stage, was submitted to continuous fasting until death; the second group, arranged in the same way, was fed once on chicken. Through this work, it was possible not only to compare the results obtained from the first group with other authors' results but, also, to analyze the dynamics of evolution, molting, longevity and the increase in insect longevity, in the second group. The average values recorded for survival time revealed statistical differences between the two groups. Among the important results detected, there is one that deserves to be emphasized: the incredible increase in longevity among insects that received only one feed - an average increase in survival time that reached 2.95 to 3.30 times in nymphs of 3rd and 4th stages, respectively. One 5th stage nymph survived for up to 350 days and the females may triplicate their survival rate, what represents an important epidemiological factor. The knowledge about this type of biological characteristic of T. vitticeps may contribute to prevent the domiciliation of this species, what seems to be incipient in some municipal districts, in Brazil

Attempts to improve xenodiagnosis: comparative test of sensibility using Rhodnius neglectus, Panstrongylus megistus, Triatoma vitticeps and Triatoma infestans in endemic areas of Brazil

From June 1984 to July 1992, 392 xenodiagnostic tests were applied on 264 patients with chronic Chagas disease from Brazilian endemic areas of Virgem da Lapa and Coronel Murta, situated in the Jequitinhonha Valley, in the State of Minas Gerais. The susceptibilities of Rhodnius neglectus, Panstrongylus megistus, Triatoma vitticeps and Triatoma infestans were compared. Most of the time nymphs (fourth instar) of each species were applied to 161 women and 103 aged between 5 and 83 years of age. The tests were preapred to compare the susceptibilities of two species at a time, using the same patients for each test. Results showed a xenopositiveness of 26.28 per cent (103 tests) being 27.98 per cent in women (68 positive in 243 applied tests) and 23.49 per cent in men (35 positive in 149 applied tests). The relative frequency of xenopositiveness displayed a great superiority of P. megistus and T. vitticeps. In tests from type I, for example, P. megistus was the unique responsable for 10.73 per cent of positive xenodiagnosis vs. only 0.98 per cent in T. infestans. Other parameters analised in this work confirm this superiority, and corroborate that T. infestans can be replaced by P. megistus and/or T. vitticeps in order to upgrade the efficacy of xenodignosis.

In vivo differentiation of Trypanosoma cruzi--1. Experimental evidence of the influence of vector species on metacyclogenesis

Vector species has not hitherto been studied as influencing metacyclogenesis of Trypanosoma cruzi, while the role of the parasite strain has been frequently stressed as of dominant importance in this process. In order to fill this gap in our knowledge, metacyclogenesis was monitored in nine triatomine species. The first part of this paper presents photographs of the main and intermediate parasite stages in each vector species studied. In the second part of the study the proportional distribution of all these forms, as seen in Giemsa stained smears is summarized, thus providing an opportunity to analyze both: the length of time between the ingestion of the blood trypomastigotes and the appearance of metacyclic forms and the rates of developmental stages leading to these latter. The most remarkable observation was that metacyclogenesis rates in vivo appear to be vector dependent, reaching 50 in Rhodnius neglectus, 37 in its congener R. prolixus and being dramatically lower in the majority of Triatoma species (5 in T. sordida, 3 in T. brasiliensis and 0 in T. pseudomaculata) at the 120th day of infection. These observations suggest that through screening of different vector species it is possible to find some that are capable of minimizing or maximizing metacyclic production.

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..

Estudos anteriores (1982, 1987) enfatizaram a superioridade das espécies vetoras silvestres às domiciliadas, como agentes do xenodiagnòstico nas infecçöes agudas e crônicas pelo isolado Y do T.cruzi. Os dados atuais estabeleceram a generalidade deste fenômeno. Por exemplo, a proporçäo de positivos nas infecçöes por "Säo Felipe" variou de 82,5 por cento a 98,3 por cento nos vetores silvestres, caindo para 42,5 por cento a 71,3 por cento em espécies domiciliadas. Ficou claro o envolvimento do isolado do T.cruzi na resposta do vetor à infecçäo crônica por T.cruzi. Por exemplo, os índices de infectividade produzidos por "Berenice", "Y", "FL" e "CL" variaram de 56,3 por cento a 88,3 por cento em P.megistus e de 28,8 por cento a 58,8 por cento em T.sordida. Sendo a resposta das mesmas espécies vetoras às infecçöes agudas pelos mesmos isolados altamente uniformes, aproximando-se a 100 por cento, sugere-se que a grande carga parasitária nas infecçöes agudas oculta as diferenças interespecíficas existentes entre os isolados do T.cruzi. A análise dos dados obtidos levou a especular que resultados adicionais de mais isolados permitiriam associar os índices de infectividade, produzidos por diferentes isolados do T.cruzi, aos perfis isoenzimáticos revelados por esses

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 Tripanosoma cruzi

É examinada e comparada a reaçäo de nove espécies vetoras da doença de Chagas à infecçäo, por sete diferentes cepas do T. cruzi (Berenice, Y, FL, CL, Säo Felipe, Colombiana e Gávea). Com base na habilidade em estabelecer e manter a infecçäo, as espécies vetoras podem ser divididas em dois grupos distintos, que diferem em suas reaçöes à infecçäo aguda por T.cruzi. Enquanto a proporçäo de insetos positivos foi baixa em domiciliados (Triatoma infestans e Triatoma dimidiata), foi alta nos considerados completamente selvagens (Rhodnius neglectus e Triatoma rubrovaria), ao serem iniciadas suas colonizaçöes no laboratório, no início da década de 70, e nos essencialmente silvestres (Panstrongylus megistus, Triatoma sordida e Triatoma pseudomaculata). Admite-se que devido à exploraçäo agropecuária e graças às campanhas de controle, os dois últimos grupos encontram mais freqüentemente condiçöes que lhes permitem maior convivência com o homem e animais domésticos. As proporçöes de positivos nas cinco últimas espécies acima citadas, cada qual infectada com uma das sete cepas do T.cruzi, quando somadas (34 "Overalls") variam de 90% a 100%, com exceçäo de 65,6% encontrada em P.megistus infectado com a cepa Gávea. A posiçäo intermediária está sendo ocupada por Triatoma brasiliensis e Rhodnius prolixus, o último alternando entre biótipo natural e artificial. Achado relevante foi a uniformidade de reaçöes dos vetores silvestres às infecçöes com, praticamente, todas as cepas do T.cruzi, sugerindo que o fator ou fatores responsáveis pela reaçäo do P.megistus à infecçäo pela cepa Y também operam nas reaçöes desta espécie com as restantes cepas, embora, várias destas tenham sido bioquimicamente diferentes entre si. A comparaçäo dos dados aqui apresentados com os relatados por outros investigadores, forma a base da discussäo sobre a superioridade de uso do D.maximus como agente no xenodiagnóstico.