Species-specific differences in the egg exochorium of the sympatric taxa Triatoma costalimai and Triatoma jatai (Hemiptera: Reduviidae: Triatominae).

Triatoma costalimai and Triatoma jatai are related species, which occur in sympatry in Paranã, Tocantins, Brazil, in rocky outcrops and in peridomicile and intradomicile environments. This study compared morphologic and morphometric aspects of the eggs of these species using optical microscopy (OM) and scanning electron microscopy (SEM). Operculum cells (OP) and egg body (EB) were drawn and photographed, their surfaces were measured, and spots were quantified. Statistical analyses were performed using ANOVA and t-tests. OM showed an egg exochorium with spots in T. costalimai and a predominance of short lines in T. jatai. We found significant differences in egg length and width, which were larger in T. costalimai. SEM analysis showed that the operculum of both species had cells with straight and/or rounded rims, with a smooth aspect, random spots, and predominantly pentagonal shape. In the EB, hexagonal cells were predominant, with indices exceeding 60% in both species. Triatoma costalimai cells were flat, with a discrete definition of the rims, whereas T. jatai cells were smooth with well-defined rims. Statistical tests showed significant differences for EB, where T. costalimai cells were larger and have more spots than T. jatai. The eggs can thus be differentiated, thereby contributing to integrative taxonomy.

The connection between Trypanosoma cruzi transmission cycles by Triatoma brasiliensis brasiliensis: A threat to human health in an area susceptible to desertification in the Seridó, Rio Grande do Norte, Brazil.

An outbreak of Chagas disease, possibly involving its vector Triatoma brasiliensis brasiliensis, was identified in the state of Rio Grande do Norte (RN). Given the historical significance of this vector in public health, the study aimed to evaluate its role in the transmission dynamics of the protozoan Trypanosoma cruzi in an area undergoing desertification in the Seridó region, RN, Brazil. We captured triatomines in sylvatic and anthropic ecotopes. Natural vector infection was determined using parasitological and molecular methods and we identified discrete typing units (DTUs) of T. cruzi by analyzing the COII gene of mtDNA, 24Sα rDNA, and mini-exon gene. Their blood meals sources were identified by amplification and sequencing of the mtDNA cytochrome b gene. A total of 952 T. b. brasiliensis were captured in peridomestic (69.9%) and sylvatic ecotopes (30.4%). A wide range of natural infection rates were observed in peridomestic (36.0% - 71.1%) and sylvatic populations (28.6% - 100.0%). We observed the circulation of TcI and TcII DTUs with a predominance of Tcl in sylvatic and peridomestic environments. Kerodon rupestris, rocky cavy (13/39), Homo sapiens, human (8/39), and Bos taurus, ox (6/39) were the most frequently detected blood meals sources. Thus, Triatoma b. brasiliensis is invading and colonizing the human dwellings. Furthermore, high levels of natural infection, coupled with the detection of TcI and TcII DTUs, and also the detection of K. rupestris and H. sapiens as blood meals sources of infected T. b. brasiliensis indicate a risk of T. cruzi transmission to human populations in areas undergoing desertification.

External female genitalia of Triatoma jatai, Triatoma costalimai and Triatoma williami (Hemiptera: Reduviidae: Triatominae).

BACKGROUND: Taxonomic identification of triatomines is generally performed based on aspects of their external morphology. However, the use of a multidisciplinary approach, considering morphological aspects of the external genitalia, morphometry, genetics, and phylogeography has been suggested, especially for similar and/or cryptic species. The rupestral species Triatoma jatai Gonçalves et al., 2013, Triatoma costalimai Verano & Galvão, 1959 and Triatoma williami Galvão et al., 1965, which are morphologically similar, have been found naturally infected with Trypanosoma cruzi (Chagas, 1909) in wild, peridomestic, and intradomestic environments, representing a risk of new outbreaks of Chagas disease. This study presents morphological description complementation of these species, with an emphasis on the structures of the female external genitalia, using scanning electron microscopy. METHODS: The females of T. jatai and T. costalimai (n = 10 of each) were captured in the Brazilian municipalities of Paranã and Aurora do Tocantins and were identified with the use of a dichotomous key for the Matogrossensis subcomplex. Females of T. williami (n = 5), were obtained from a laboratory colony. The females were cut transversely at the sixth abdominal segment and examined under scanning electron microscopy (SEM) at the Oswaldo Cruz/Fiocruz Institute Electronic Microscopy Platform. RESULTS: It was possible to differentiate the three species based on the characteristics of urotergites VII, VIII and IX and urosternite VII, as well as the genital plaques, gonocoxites, and gonapophyses. To our knowledge, morphological differences in the spines present on gonapophysis 8 in triatomines are described here for the first time. CONCLUSIONS: The results show that external genitalia of females are useful structures to differentiate T. costalimai, T. jatai and T. williami. SEM analysis contributes to and corroborates, together with other tools morphological and molecular, the distinction of the three species.

Does antennal sensilla pattern of different populations of Triatoma maculata (Hemiptera: Reduviidae) reveal phenotypic variability?

BACKGROUND: In Brazil, Triatoma maculata is only found in the State of Roraima and is a vector of Trypanosoma cruzi, the etiological agent of Chagas disease. It occurs in wild, peridomestic and domestic habitats, with an urban infestation in Boa Vista, the capital of this Brazilian state. The aim of this study was to assess the morphological variability of the T. maculata antennal phenotype in three populations of Roraima State, using the antennal sensilla pattern analyzed under optical microscopy. METHODS: The number and distribution of four antennal sensilla types (bristles, thin and thick walled trichoidea, and basiconic) of three Brazilian populations of T. maculata from Roraima State were compared. Univariate and multivariate analyses were performed. RESULTS: The antenna of T. maculata presented the four types of sensilla. According to the density and distribution of the antennal sensilla characteristics, the multivariate analyses showed that the laboratory population is morphologically structured. Urban specimens showed a pronounced phenotypic variability. The main differences were observed in the pedicel segment, and between males and females. CONCLUSIONS: We determined the antennal phenotype in three Roraima populations of T. maculata. These results support the idea that the patterns of antennal sensilla are sensitive markers for distinct populations in the Triatominae. The infestations of T. maculata in different habitats reinforces the ability of this vector to become adapted to a variety of environments, which, could have eco-epidemiological implications for the T. cruzi transmission that are still not well understood.

Triatoma costalimai Naturally Infected by Trypanosoma cruzi: A Public Health Concern.

The rupestrian Triatoma costalimai species has been found infected by Trypanosoma cruzi in wild, peridomicile, and intradomicile environments in the municipality of Aurora do Tocantins, Tocantins, Brazil. Proximity between rock outcrops increases the risk of vector transmission of Chagas disease via this species. This work describes a focus of colonization by T. costalimai specimens infected by T. cruzi in rock outcrops located in an urban area in this municipality. Parasitological examination of feces from the collected specimens, axenic cultivation of T. cruzi-positive samples, and genetic characterization of the isolates were performed. Nymph and adult specimens were collected with a high infection prevalence (64.5%) for T. cruzi discrete type unit (DTU I). Participation of the T. costalimai species in the wild cycle of T. cruzi in rock outcrops located in an urban area demonstrates the need for entomological surveillance and control of vector transmission of Chagas disease in the municipality of Aurora do Tocantins, Tocantins.

Diversity of Mosquitoes At the Itatiaia National Park, State of Rio De Janeiro, Brazil.

A temporal observational study was conducted with the aim of characterizing Culicidae richness, diversity, abundance, and species similarity in fragments of Atlantic Forest within the Itatiaia National Park, state of Rio de Janeiro, Brazil. A total of 31 mosquito species were collected in 2 sampling stations, during the day and night, and the abundance index and dominance coefficient for each species were calculated. The results showed that the culicid density was higher during the day, and the time preference was directly influenced by the climate variables analyzed, which acted as limiting factors for mosquito occurrence. The mosquito fauna at the Itatiaia National Park presented high richness, including some epidemiologically important species: Anopheles cruzii, Haemagogus leucocelaenus, Aedes scapularis, and Sabethes identicus. Studies of this region supply information on biodiversity, and consequently help to understand epidemiological aspects of potential pathogen vector species.

Triatoma maculata colonises urban domicilies in Boa Vista, Roraima, Brazil.

During a medical entomology course in Boa Vista, Roraima, colonies of Triatoma maculata closely associated with pigeon nests were observed in concrete air-conditioner box located on the external plastered and cemented walls of a modern brick-built apartment block. In only one eight-hole ceramic brick, located inside one air-conditioner box, 127 specimens of T. maculata were collected. T. maculata is a recognised vector of Trypanosoma cruzi in the surrounding area and its domiciliation increases the risk of Chagas disease transmission.

Mitochondrial Genes Reveal Triatoma jatai as a Sister Species to Triatoma costalimai (Reduviidae: Triatominae).

Triatoma jatai was described using a set of morphological structures from specimens collected in Paranã municipality of Tocantins State, Brazil. Under a Bayesian framework and using two mitochondrial genes (16S and COI), phylogenetic analysis recovered T. jatai as a sister species to Triatoma costalimai with higher genetic distances than between other well-recognized species. Our results agree with previous suggestions based on morphometric analysis. In the light of the non-monophyly of Matogrossensis subcomplex, the inclusion of T. jatai shall be considered for reevaluating this group.

Triatoma jatai sp. nov. in the state of Tocantins, Brazil (Hemiptera: Reduviidae: Triatominae).

Triatoma jatai sp. nov. is the first new species of triatomine to be described in the state of Tocantins, in the northern region of Brazil. It was caught on rock outcrops in the wild environment and, more recently, invading homes. While T. jatai sp. nov. is morphologically similar to Triatoma costalimai, it is distinguished by its general colouring, differences in the blotches on the connexivum, wing size in females and external structures of the male genitalia. The type series has been deposited in the Entomological Collection and Herman Lent Collection, Oswaldo Cruz Institute-Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.

Triatoma jatai sp. nov. in the state of Tocantins, Brazil (Hemiptera: Reduviidae: Triatominae)

Triatoma jatai sp. nov. is the first new species of triatomine to be described in the state of Tocantins, in the northern region of Brazil. It was caught on rock outcrops in the wild environment and, more recently, invading homes. While T. jatai sp. nov. is morphologically similar to Triatoma costalimai, it is distinguished by its general colouring, differences in the blotches on the connexivum, wing size in females and external structures of the male genitalia. The type series has been deposited in the Entomological Collection and Herman Lent Collection, Oswaldo Cruz Institute-Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.

Biodiversity and times of activity of mosquitoes (Diptera: Culicidae) in the biome of the Atlantic Forest in the State of Rio de Janeiro, Brazil.

A temporal observational study on culicid entomofauna was conducted in a region characterized as a fragment of the Atlantic Forest that forms the Tinguá Biological Reserve in the State of Rio de Janeiro. This investigation was performed with the aim of analyzing the influence of climatic factors (temperature and relative air humidity) on the activity levels at different times of the day among mosquito species within the ecosystems that form the Tinguá Biological Reserve. The abundance index and dominance coefficient were calculated in relation to 61 mosquito species that were caught at four sampling sites, in the mornings, afternoons, and evenings. The results revealed that culicid species were distributed with greater incidence during the two diurnal periods and that their preference for times of the day was directly influenced by the climatic variables analyzed. The latter acted as limiting factors for occurrences of mosquito species.

Sylvatic and peridomestic populations of Triatoma pseudomaculata are not significantly structured by habitat, as revealed by two genetic markers.

Chagas disease remains a public health concern in Brazil and other Latin American countries, mainly due to the potential domiciliation of native triatomine species. We analyzed the genetic variability of Triatoma pseudomaculata in sylvatic and peridomestic ecotopes throughout three localities in the northeastern state of Bahia, Brazil. We studied polymorphisms generated by random amplified polymorphic DNA (RAPD) and isoenzyme electrophoresis analyses. Based on RAPD analysis, each specimen was assigned to one of three genetic clusters. Although all sylvatic specimens from one locality were grouped into the same cluster, sylvatic and peridomestic specimens from the other two localities were broadly distributed between the remaining two clusters, suggesting that geographic population structuring was not occurring. Furthermore, isoenzyme analysis suggested that distinct populations were in Hardy-Weinberg equilibrium. Low statistical values for Wright's Fst index also supported the absence of population structuring and suggested the occurrence of panmixia. We conclude that genetic flow occurs between sylvatic and peridomestic T. pseudomaculata populations, probably as a consequence of passive and active dispersion of the insects, associated with deforestation and anthropic transformations.

Comparative biology of the two sister species of Triatominae (Hemiptera: Reduviidae).

INTRODUCTION: Triatoma pseudomaculata and T. wygodzinskyi (Hemiptera: Reduviidae: Triatominae) are two Brazilian vectors of Chagas disease. The first is an arboricolous species in sylvatic environment and considered a vector of T. cruzi in peridomestic structures; the second, a rupicolous species in the wild environment of no epidemiological importance. In order to test the assumption that sister species share biological traits, comparative studies of their development cycle and blood ingestion were conducted. METHODS: Eggs laid by five field females of each species were randomly selected. The nymphs were observed daily and fed on mice weekly. The time required to pass through the different stages to adulthood was recorded in days. The triatomines were weighed individually before and after feeding. The mortality rate according to each nymphal stage was calculated. RESULTS AND CONCLUSIONS: Analysis of the results shows that they display only minor biological differences even though they exhibit a distinct ecology. This suggests that the biological traits are important criteria to determine the relationship between species.

Comparative biology of the two sister species of Triatominae (Hemiptera: Reduviidae) / Biologia comparativa de duas espécies irmãs de Triatominae (Hemiptera: Reduviidae)

INTRODUCTION: Triatoma pseudomaculata and T. wygodzinskyi (Hemiptera: Reduviidae: Triatominae) are two Brazilian vectors of Chagas disease. The first is an arboricolous species in sylvatic environment and considered a vector of T. cruzi in peridomestic structures; the second, a rupicolous species in the wild environment of no epidemiological importance. In order to test the assumption that sister species share biological traits, comparative studies of their development cycle and blood ingestion were conducted. METHODS: Eggs laid by five field females of each species were randomly selected. The nymphs were observed daily and fed on mice weekly. The time required to pass through the different stages to adulthood was recorded in days. The triatomines were weighed individually before and after feeding. The mortality rate according to each nymphal stage was calculated. RESULTS AND CONCLUSIONS: Analysis of the results shows that they display only minor biological differences even though they exhibit a distinct ecology. This suggests that the biological traits are important criteria to determine the relationship between species.

INTRODUÇÃO: Triatoma pseudomaculata e T. wygodzinskyi (Hemiptera: Reduviidae: Triatominae) são dois vetores Brasileiros da doença de Chagas. A primeira é uma espécie arborícola em ambiente silvestre e considerada vetor do T. cruzi em estruturas peridomesticas. A segunda é rupícola em ambientes silvestres e sem importância epidemiológica. Com o objetivo de testar a hipótese que espécies irmãs compartilham características semelhantes, realizamos um estudo comparativo do ciclo biológico e ingesta alimentar. MÉTODOS: Ovos pertencentes a cinco fêmeas de cada espécie provenientes do campo foram selecionados aleatoriamente. As ninfas foram observadas diariamente e alimentadas com camundongos semanalmente. O tempo requerido para passar até o estágio adulto foi registrado em dias. Os triatomíneos foram pesados individualmente antes e depois da alimentação. A mortalidade de cada estágio foi calculada. RESULTADOS E CONCLUSÕES: Analise dos resultados mostram que eles exibem pequenas diferencias biológicas apesar de apresentarem diferenças marcantes quanto a sua ecologia. Isso sugere que as características biológicos são critérios importantes para determinar a relação entre as espécies.

Phylogenetic and phenotypic relationships among Triatoma carcavalloi (Hemiptera: Reduviidae: Triatominae) and related species collected in domiciles in Rio Grande do Sul State, Brazil.

Triatoma carcavalloi is considered a rare Chagas disease vector often collected inside domiciles in Rio Grande do Sul State. In this Brazilian state, T. carcavalloi has been collected in the same ecotope (rock piles) with two other species (T. rubrovaria and T. circummaculata), with which it also shares morphological characteristics. Previous morphological studies placed T. carcavalloi in the same species complex ("infestans complex") and subcomplex ("rubrovaria subcomplex") as T. rubrovaria, whereas T. circummaculata was placed in the "circummaculata complex." The phylogeny of a group composed of 16 species of triatomines was reevaluated with the inclusion of T. carcavalloi by Bayesian analysis using mtDNA sequences of subunits 12S and 16S of the ribosomal RNA, and the cytochrome oxidase I (COI) genes. The phenotypic relationship among T. carcavalloi and related triatomines was also inferred from morphometrics. Phylogenetic results indicate that T. carcavalloi is a sister species of T. rubrovaria, and both were recovered as closely related to T. circummaculata. Morphometric studies confirmed the closeness among T. carcavalloi, T. rubrovaria, and T. circummaculata, prompting the placement of the latter species in the "infestans complex" and "rubrovaria subcomplex."

Behavioral plasticity of Triatominae related to habitat selection in northeast Brazil.

Triatoma pseudomaculata Corrêa and Espínola, 1964 and Triatoma juazeirensis Costa and Felix, 2007 ( = T. brasiliensis Neiva, 1911 [part]), are sylvatic vectors of Trypanosoma cruzi (Chagas, 1909), the causative agent of Chagas disease, in northeast Brazil-especially in the caatinga region. In an area of caatinga in the State of Bahia, we compared the wild and peridomestic habitats of these two species of Triatominae to assess their behavioral plasticity in relation to habitat selection in different environments. In the sylvatic environment, the habitat of these two species is never shared. T. pseudomaculata is found in trees and bird nests, but without apparent preference for any particular tree species. In contrast, T. juazeirensis is exclusively rupicolous (found among rocks). Both species invade peridomestic structures but do not display a significant ability to colonize human dwellings. In the peridomestic area, they are highly adaptable to different habitats and can occupy substrates that they do not colonize in the sylvatic environment. This behavioral plasticity seems to be more striking in T. juazeirensis-rupicolous in sylvatic environments but colonizing wooden structures in the peridomicile in >80% of cases.

Ecology of Anopheline (Diptera, Culicidae), malaria vectors around the Serra da Mesa Reservoir, State of Goiás, Brazil. 1--Frequency and climatic factors.

The ecology of anopheline species (Diptera, Culicidae) was studied in the vicinity of the Serra da Mesa Reservoir, State of Goiás, Brazil. Climatic factors and frequency of anopheline populations were analyzed. Bimonthly human-bait and Shannon trap captures were conducted for 36 consecutive months (January 1997 through December 1999). A total of 5,205 adult anophelines belonging to five species were collected. Anopheles darlingi was the most frequently collected anopheline (61.4%), followed by An. albitarsis s.l. (35.4%), An. triannulatus. (2.5%), An. oswaldoi (0.4%), and An. evansae (0.2%). The water level and vegetation along the banks of the reservoir were crucial to the frequency of the various anopheline species. Climatic factors had a secondary influence. The reservoir's water-level stability, increased frequency of An. darlingi, and the arrival of gold prospectors were responsible for the increase in malaria cases.

Ecology of Anopheline (Diptera, Culicidae), malaria vectors around the Serra da Mesa Reservoir, State of Goiás, Brazil: 1 - Frequency and climatic factors

Foram estudados aspectos da ecologia de anofelinos (Diptera, Culicidae) em áreas da Usina Hidrelétrica de Serra da Mesa, Estado de Goiás, Brasil. Foram analisados a incidência das populações de anofelinos e os fatores climáticos. Foram realizadas capturas bimestrais, em isca humana e armadilha Shannon, ao longo de 36 meses consecutivos (janeiro de 1997 a dezembro de 1999). Capturamos um total de 5.205 anofelinos adultos, pertencentes a cinco espécies. Anopheles darlingi foi o anofelino mais freqüente (61,4 por cento), seguido de An. albitarsis s.l. (35,4 por cento), An. triannulatus (2,5 por cento), An. oswaldoi (0,4 por cento) e An. evansae (0,2 por cento). O nível da água e a vegetação às margens do reservatório foram determinantes para a freqüência das espécies de anofelinos. Os fatores climáticos tiveram influência secundária. A estabilidade do reservatório, o aumento na freqüência do An. darlingi e a chegada de garimpeiros foram responsáveis pelos casos de malária diagnosticados.

[Scanning electron microscopy of eggs of Haemagogus leucocelaenus (Diptera: Culicidae)]. / Microscopia eletrônica de varredura de ovos de Haemagogus leucocelaenus (Diptera: Culicidae).

OBJECTIVE: To observe morphological details of the eggs of Haemagogus (Conopostegus) leucocelaenus, seen for the first time via scanning electron microscopy (SEM), with morphometric analysis of the main structures. METHODS: Eggs of Hg. leucocelaenus were obtained from females captured in the Biological Reserve of Tinguá, State of Rio de Janeiro. Some of the eggs were kept for hatching and others underwent processing for scanning electron microscopy studies. Three eggs were submitted to morphometric analysis. The material was fixed in 2.5% glutaraldehyde and postfixed in 1% osmium tetroxide, both in 0.1M, pH 7.2 sodium cacodylate buffer, then dehydrated in ethanol and dried using the critical point method. This was then set up on metallic supports, covered with gold and observed using the Jeol 5310 scanning electron microscope. Measurements were made with the aid of the Semafore analysis software coupled to the electron microscope. RESULTS: The eggs presented elliptical outlines of approximately 574 mm in length and 169 mm in width, with an egg index (l/w ratio) of 3.39 mm. The exochorion was extremely regular and had ornamentation that was usually hexagonal but sometimes pentagonal. Tubercles were observed on the chorionic cells, symmetrically arranged in relation to the longitudinal axis. Inside the cells, there were smaller, individualized tubercles, some arranged peripherally and others grouped to a greater or lesser extent in the center. The surface of the chorionic reticulum did not present rugosity. The micropylar apparatus was formed by a prominent continuous collar of 8.32 mm in thickness, with a slightly irregular surface. The micropylar disk was very evident, and was continuous with the collar. The micropyle was seen at the center of this disk, measuring 1.6 mm and with a micropylar apparatus of 27.3 mm in diameter. CONCLUSIONS: The ornamentation of the exochorion presents differences in relation to the tubercles of chorionic cells and the external chorionic reticulum between the eggs of Hg. Leucocelaenus, in comparison with the eggs of Hg. janthinomys and Hg. equinus, and also in relation to those of Aedes aegypti, Ae. albopictus and Ae. bahamensis. In various aspects, the eggs of Hg. leucocelaenus have more resemblance to those of Hg. Equinus than those of Hg. janthinomys, with greater differences presented in relation to the eggs of Hg. spegazzinii and Hg. lucifer.

Microscopia eletrônica de varredura de ovos de Haemagogus leucocelaenus (Diptera: Culicidae) / Scanning electron microscopy of eggs of Haemagogus leucocelaenus (Diptera: Culicidae)

OBJETIVO: Observar detalhes morfológicos de ovos de Haemagogus leucocelaenus visualizados pela primeira vez por microscopia eletrônica de varredura (MEV) e realizar morfometria das principais estruturas. MÉTODOS: Foram utilizados ovos de Hg. leucocelaenus provenientes de fêmeas capturadas na Reserva Biológica do Tinguá, RJ, sendo parte destinada à eclosäo e outra ao processamento de MEV, dos quais três foram submetidos à análise morfométrica. O material foi fixado em glutaraldeído 2,5 por cento e pós-fixado em tetróxido de ósmio 1 por cento, ambos em tampäo cacodilato de sódio 0.1M, pH 7.2, processado e observado ao MEV Jeol 5310. Medições foram realizadas com o auxílio do software de análise Semafore. RESULTADOS: Os ovos apresentaram contorno elíptico com aproximadamente 574 æm de comprimento e 169 æm de largura, sendo o índice do ovo (l/wratio) 3,39 æm. O exocório é extremamente regular, possuindo ornamentaçäo hexagonal e algumas vezes pentagonal. Nas células coriônicas, observaram-se tubérculos simetricamente dispostos com relaçäo ao eixo longitudinal, e, no interior delas, tubérculos menores, individualizados, dispostos na periferia, e poucos agrupados no centro. A superfície do retículo coriônico näo apresentou rugosidades. O aparelho micropilar apresenta colar proeminente, contínuo, com disco micropilar bem evidente. CONCLUSÕES: A ornamentaçäo do exocório apresenta diferenças em relaçäo aos tubérculos das células coriônicas e ao retículo coriônico externo entre os ovos de Hg. leucocelaenus comparados aos ovos de Hg. janthinomys e Hg. equinus, bem como com relaçäo aos de Aedes aegypti, Ae. albopictus e Ae bahamensis.