Biogeography of Triatominae (Hemiptera: Reduviidae) in Ecuador: implications for the design of control strategies

Chagas disease control strategies strongly depend on the triatomine vector species involved in Trypanosoma cruzi transmission within each area. Here we report the results of the identification of specimens belonging to various species of Triatominae captured in Ecuador (15 species from 17 provinces) and deposited in the entomological collections of the Catholic University of Ecuador (Quito), Instituto Oswaldo Cruz (Brazil), the Natural History Museum London (UK), the London School of Hygiene and Tropical Medicine (UK), the National Institute of Hygiene (Quito), and the Vozandes Hospital (Quito). A critical review of published information and new field records are presented. We analysed these data in relation to the life zones where triatomines occur (11 life zones, excluding those over 2,200 m altitude), and provide biogeographical maps for each species. These records are discussed in terms of epidemiological significance and design of control strategies. Findings relevant to the control of the main vector species are emphasised. Different lines of evidence suggest that Triatoma dimidiata is not native to Ecuador-Peru, and that synanthropic populations of Rhodnius ecuadoriensis in southern Ecuador-northern Peru might be isolated from their sylvatic conspecifics. Local eradication of T. dimidiata and these R. ecuadoriensis populations might therefore be attainable. However, the presence of a wide variety of native species indicates the necessity for a strong longitudinal surveillance system.

Differentiation and genetic analysis of Rhodnius prolixus and Rhodnius colombiensis by rDNA and RAPD amplification

Domiciliated Rhodnius prolixus and sylvatic R. colombiensis were analyzed in order to confirm their genetic divergence and verify the risk that the latter represents in the domiciliation process, and to provide tools for identifying the sources of possible reinfestation by triatomines in human dwellings allowing control programs to be undertaken. Comparison of random amplified polymorphic DNA amplification patterns and cluster analysis suggests reproductive discontinuity between the two species. The calculated statistical F value of 0.24 and effective migration rate of 0.6 individuals per generation are insufficient to maintain genetic homogeneity between them and confirm the absence of present genetic flow. R. colombiensis presents higher intrapopulation variability. Polymerase chain reaction of ribosomal DNA supports these findings. The low genetic flow between the two species implies that R. colombiensis do not represent an epidemiological risk for the domiciliary transmission of Trypanosoma cruzi in the Tolima Department. The lower variability of the domiciliated R. prolixus could result in greater susceptibility to the use of pesticides in control programs.

The Triatomines of Northern Peru, with Emphasis on the Ecology and Infection by Trypanosomes of Rhodnius ecuadoriensis (Triatominae)

Information on the distribution and synanthropic behaviour of triatomines is essential for Chagas disease vector control. This work summarises such information from northern Peru, and presents new data on Rhodnius ecuadoriensis - an important local vector infesting 10-35% of dwellings in some zones. Three species are strongly synanthropic and may be suitable targets for chemical control of domestic/peridomestic bug populations. Panstrongylus herreri, the main domestic vector in the area, is probably present in sylvatic ecotopes in the Marañón river system. R. ecuadoriensis and Triatoma dimidiata seem exclusively domestic; biogeographical and ecological data suggest they might have spread in association with humans in northern Peru. Confirmation of this hypothesis would result in a local eradication strategy being recommended. Presence of trypanosome natural infection was assessed in 257 R. ecuadoriensis; Trypanosoma rangeli was detected in 4% of bugs. Six further triatomine species are potential disease vectors in the region (T. carrioni, P. chinai, P. rufotuberculatus, P. geniculatus, R. pictipes, and R. robustus), whilst Eratyrus mucronatus, E. cuspidatus, Cavernicola pilosa, Hermanlentia matsunoi, and Belminus peruvianus have little or no epidemiological significance. A strong community-based entomological surveillance system and collaboration with Ecuadorian public health authorities and researchers are recommended.

Oral Susceptibility to Yellow Fever Virus of Aedes aegypti from Brazil

The oral susceptibility to yellow fever virus was evaluated in 23 Aedes aegypti samples from Brazil. Six Ae. aegypti samples from Africa, America and Asia were also tested for comparison. Mosquito samples from Asia showed the highest infection rates. Infection rates for the Brazilian Ae. aegypti reached 48.6%, but were under 13% in 60% of sample tested. We concluded that although the low infection rates estimated for some Brazilian mosquito samples may not favor the establishment of urban cycle of yellow fever in some parts of the country, the founding of Ae. aegypti of noteworthy susceptibility to the virus in cities located in endemic and transition areas of sylvatic yellow fever, do pose a threat of the re-emergence of the urban transmission of the disease in Brazil.

Geometric morphometric differences between Panstrongylus geniculatus from field and laboratory

The finding of Panstrongylus geniculatus nymphs inside a house in northeastern Antioquia, Colombia, and the reports related to their increasing presence in homes suggest the need for surveillance methods for monitoring the invasion processes. We analyzed the morphological differences between a wild population and its laboratory descendants, using the techniques of geometric morphometry, with the idea that such differences might parallel those between sylvatic and synanthropic populations. The analyses over five generations showed differences in size but not in shape. Head size and wing size were both reduced from sylvatic to laboratory populations, but the decrease in head size occurred only up to the second generation while the decrease in wing size proceeded up to the fifth generation. In contrast, although a decrease in sexual size dimorphism has been proposed as a marker of colonization in human dwellings, we did not detect any significant loss of dimorphism between sexes of P. geniculatus over the five generations studied. We conclude that size changes may have a physiological origin in response to a change of ecotopes, but more than five generations may be required for the expression of permanent morphological markers of human dwellings colonization.

Triatoma rubrovaria (Blanchard, 1843) (Hemiptera, Reduviidae, Triatominae) II: trophic resources and ecological observations of five populations collected in the State of Rio Grande do Sul, Brazil

Triatoma rubrovaria has become the most frequently captured triatomine species after the control of T. infestans in the State of Rio Grande do Sul (RS), Brazil. Isoenzymatic and chromatic studies indicate the existence of, at least, two distinct phenotypic patterns of T. rubrovaria in RS. The geographic variation noted through molecular tools may also result in distinct profiles of vectorial potentiality. In order to enhance our understanding of the bionomic knowledge of T. rubrovaria separate batches of the species were collected from different municipalities of RS distant from 72 to 332 km: Santana do Livramento (natural ecotope), Santana do Livramento (artificial ecotope), Santiago (natural ecotope), Canguçu (peridomicile) and Encruzilhada do Sul (natural ecotope). A total of 285 specimens were collected, 85 specimens kept sufficient fecal material in their guts for the precipitin analysis. The results indicated the food eclecticism for this species and the anti-rodent serum showed the highest positivity in most localities. From the total of analyzed samples, only 1.3% of unique positivity for human blood was registered, all of them for Santiago population. This reactivity to human blood may be associated to pastures activities in the field.

Estudo morfométrico do gênero Panstrongylus Berg, 1879 (Hemiptera, Reduviidae, Triatominae)

The subfamily Triatominae is actually represented by 137 species distributed among 6 tribes and 19 genera. Within this subfamily, the genus Panstrongylus, Berg 1879, is composed by 13 species widespread in sylvatic, peridomestic, and domestic habitats of Neotropical regions. These species are vectors of Chagas disease and consequently are found associated with its main hosts, such as birds and mammals. Interest in species of this genus has been increasing in the last few years. Reports of several authors indicate these Triatominae to invade and colonize houses, increasing their epidemiological significance. Morphometry was used in this study to investigate correlations among possible closely related species. We measured 224 specimens among 13 species through a set of metric variables of the head. The results indicated that the genus Panstrongylus seems to be homogeneous since 10 of the 14 species were shown to be closely related.

Re-infestation of houses by Triatoma dimidiata after intra-domicile insecticide application in the Yucatán peninsula, Mexico

In most countries, Chagas disease transmission control remains based on domestic insecticide application. We thus evaluated the efficacy of intra-domicile cyfluthrin spraying for the control of Triatoma dimidiata, the only Chagas disease vector in the Yucatán peninsula, Mexico, and monitored potential re-infestation every 15 days for up to 9 months. We found that there was a re-infestation of houses by adult bugs starting 4 months after insecticide application, possibly from sylvatic/peridomicile areas. This points out the need to take into account the potential dispersal of sylvatic/peridomestic adult bugs into the domiciles as well as continuity action for an effective vector control.

Isoenzimatic analysis of four Anopheles (Kerteszia) cruzii (Diptera: Culicidae) populations of Brazil

Anopheles cruzii is a small sylvatic mosquito and primary human Plasmodium vector in Southern Brazil. The distribution of this bromeliad-breeding mosquito follows the Atlantic forest coastal distribution, where bromeliads are abundant. Morphological, genetic, and molecular polymorphisms among different populations have been reported and it has recently been suggested that An. cruzii is a complex of cryptic species. The aim of this work is to analyze the gene flow between different populations of An. cruzii collected in four localities within the geographic distribution range of the species, and to examine if An. cruzii is a complex of cryptic species. The genetic distances show that populations of the states of Santa Catarina, São Paulo, and Rio de Janeiro are genetically closer (0.032 to 0.083) than populations of Bahia (0.364 to 0.853) based on profiles from 10 distinct isoenzyme loci. The Fst was lower (0.077) when the Bahia population was excluded than when it was included (0.300) in the analyses. The inferred number of migrants per generation was 2.99 individuals among populations from the states of Santa Catarina, São Paulo, and Rio de Janeiro and 0.58 migrants per generation among all populations. Results suggest that An. cruzii is a complex of species and that the specimes of state of Bahia can be considered as belonging to a species that is distinct from other three closely-related populations studied.

Ciclo biológico em laboratório de Rhodnius brethesi Matta, 1919 (Hemiptera, Reduviidae, Triatominae), potencial vetor silvestre da doença de Chagas na Amazônia

R. brethesi is a sylvatic species from the Amazon region; it has been incriminated as responsible for the transmission of Chagas disease in collectors of piaçaba in this region. The aim of present study was to investigate the efficiency of these insects as potential vectors of Trypanosoma cruzi. Aspects related with feeding and defecation patterns, life time, and mortality had been observed in each instar of R. brethesi. We use 5th instar nymphs to get adults virgins, after the moulting 3 groups with 6 females and 2 males each were created to obtain eggs. After hatching, 1st instar nymphs had been weighed and kept in bottles until the next moult. Insects were fed once a week in mice. Results showed that the average period of incubation was 17 days, the number of blood meal was increasing from the 1st to the 5th instar nymph with 7 (average) to become adult, a significative numbers of the defecations occurring immediately after the bloodmeals. The total percentual of mortality was 16%. This results suggests that this species presents a good exploitation of blood meals and a brief nymphal development in laboratory conditions reflecting its behavior in sylvatic environments.

Peridomiciliary colonies of Triatoma vitticeps (Stal, 1859) (Hemiptera, Reduviidae, Triatominae) infected with Trypanosoma cruzi in rural areas of the state of Espírito Santo, Brazil

In Brazil, the colonization of human dwellings by triatomines occurs in areas with native vegetation of the caatinga or cerrado types. In areas of Atlantic forest such as in the Brazilian state of Espírito Santo, there are no species adapted to live in human habitations. The few autochthonous cases of Chagas disease encountered in Espírito Santo have been attributed to adult specimens of Triatoma vitticeps that invade houses from forest remnants. In recent years, the entomology unit of the Espírito Santo State Health Secretariat has recorded nymphs infected with flagellates similar to Trypanosoma cruzi in rural localities. Entomological surveys were carried out in the residences and outbuildings in which the insects were found, and serological examinations for Chagas disease performed on the inhabitants. Four colonies were found, all associated with nests of opossums (Didelphis aurita), 111 specimens of T. vitticeps, and 159 eggs being collected. All the triatomines presented flagellates in their frass. Mice inoculated with the faeces presented trypomastigotes in the circulating blood and groups of amastigotes in the cardiac muscle fibres. Serological tests performed on the inhabitants were negative for T. cruzi. Even with the intense devastation of the forest in Espírito Santo, there are no indications of change in the sylvatic habits of T. vitticeps. Colonies of this insect associated with opossum nests would indicate an expansion of the sylvatic environment into the peridomicile.

Participation of cytokines in the necrotic-inflammatory lesions in the heart and skeletal muscles of Calomys callosus infected with Trypanosoma cruzi

Calomys callosus, a sylvatic reservoir of Trypanosoma cruzi, when infected with the Colombian strain (Biodeme Type III, T. cruzi I ) develops necrotic-inflammatory lesions and intense early fibrogenesis in the heart and skeletal muscles, that spontaneously regress. Participation of pro-inflammatory and pro-fibrogenic cytokines, such as tumor necrosis factor-alpha (TNF-alpha), gamma interferon (IFN-gamma) , and tumor growth factor-beta (TGF-beta), in the pathogenesis of the lesions is herein studied. Eighty C. callosus weighing 20 to 30 g were used. Seventy of them were inoculated with the Colombian strain (10(5) blood forms) and 10 were maintained as intact non-infected controls. After infection, C. callosus were sacrificed at different time-points from 15 to 70 days. The heart and skeletal muscle were processed for histopathology and cryopreserved for immunohistochemistry. Early necrotic lesions of parasitized skeletal muscle and myocardium with intense inflammatory lesions were present. Search for the in situ presence of TNF-alpha and IFN-gamma, was performed using rat-IgG anti-mouse antibodies against these cytokines. For the in situ search of TGF-beta, rabbit IgG anti-mouse antibodies were used. Immunolabeling of the cytokines in tissues of infected C. callosus was successful. The cytokines TNF-alpha, IFN-gamma , and TGF-beta were detected in the cytoplasm of macrophages and in the necrotic material from 15 to 45 days post-infection, decreasing their intensity until complete disappearance by the 65th day, which correlated with subsiding histopathological lesions. These findings suggest the participation of these cytokines in the control of parasite multiplication, in the development of an early fibrogenesis and in the regression of fibrotic-inflammatory lesions observed in C. callosus.

Dynamics of feeding and defecation in Triatoma vitticeps (Stal, 1859) (Hemiptera, Reduviidae, Triatominae) and its potential in the transmission of Trypanosoma cruzi

Adults of Triatoma vitticeps infected by flagellates similar to Trypanosoma cruzi are frequently captured by the inhabitants of rural areas in the Brazilian state of Espírito Santo. The dynamics of feeding and defecation were observed in three groups of adult triatomines, consisting of sylvatic T. vitticeps and laboratory-reared specimens of this species and T. infestans. Triatomines were observed from the moment they were presented with an immobilized chicken as a bloodmeal source until 240 min after feeding had ended. Mean times between the end of feeding and defecation for T. infestans, wild T. vitticeps and laboratory-reared specimens of the latter species were 1.2, 21.1, and 64 min respectively. All T. infestans defecated within 10 min of feeding, while only 29.9 of wild and 52.8% laboratory-reared specimens of T. vitticeps did so within this period. These results may explain the low efficiency of T. vitticeps in T. cruzi transmission to man. The shorter time between feeding and defecation in laboratory-reared T. vitticeps may indicate a change in behaviour of this species as a result of adaptation to an artificial environment.

Triatoma juazeirensis sp. nov. from the state of Bahia, Northeastern Brazil (Hemiptera: Reduviidae: Triatominae)

Triatoma juazeirensis, a new triatomine species from the state of Bahia, Northeastern Brazil, is described. The new species is found among rocks in sylvatic environment and in the peridomicile. Type specimens were deposited in the Entomological Collection of Oswaldo Cruz Institute-Fiocruz, Museum of Zoology of University of São Paulo, and Florida Museum of Natural History. T. juazeirensis can be distinguished from the other members of the T. brasiliensis species complex mainly by the overall color of the pronotum, which is dark, and by the entirely dark femora.

Chagas disease in the Amazon Region

The risk that Chagas disease becomes established as a major endemic threat in Amazonia (the world's largest tropical biome, today inhabited by over 30 million people) relates to a complex set of interacting biological and social determinants. These include intense immigration from endemic areas (possibly introducing parasites and vectors), extensive landscape transformation with uncontrolled deforestation, and the great diversity of wild Trypanosoma cruzi reservoir hosts and vectors (25 species in nine genera), which maintain intense sylvatic transmission cycles. Invasion of houses by adventitious vectors (with infection rates > 60%) is common, and focal adaptation of native triatomines to artificial structures has been reported. Both acute (~ 500) and chronic cases of autochthonous human Chagas disease have been documented beyond doubt in the region. Continuous, low-intensity transmission seems to occur throughout the Amazon, and generates a hypoendemic pattern with seropositivity rates of ~ 1-3%. Discrete foci also exist in which transmission is more intense (e.g., in localized outbreaks probably linked to oral transmission) and prevalence rates higher. Early detection-treatment of acute cases is crucial for avoiding further dispersion of endemic transmission of Chagas disease in Amazonia, and will require the involvement of malaria control and primary health care systems. Comprehensive eco-epidemiological research, including prevalence surveys or the characterization of transmission dynamics in different ecological settings, is still needed. The International Initiative for Chagas Disesae Surveillance and Prevention in the Amazon provides the framework for building up the political and scientific cooperation networks required to confront the challenge of preventing Chagas disease in Amazonia.