A New Record of the Introduced Species Triatoma infestans (Hemiptera: Reduviidae) in Mexico.

Here, we report a new record of Triatoma infestans (Klug) in Mexico after 50 years and provide a brief description of the discovery area. Fifty-nine specimens (71.2% adults) of the introduced species were collected from the peridomestic areas of a single house in the port of Manzanillo in the state of Colima, Mexico. Thirty-one specimens (52.5%) were collected from the exterior walls of the house and were apparently attracted to light. The other specimens (47.5%) were associated with chickens. No specimen was infected with Trypanosoma cruzi Chagas, the causative agent of Chagas disease, possibly because they were feeding on chickens. We speculate that the introduced species travelled from South America to Mexico via seed shipment in a twenty-foot equivalent unit (TEU) maritime container. Because Mexican phytosanitary regulations demand only the cargo to be inspected, the triatomines could have escaped notice during inspection. Subsequently, as the cargo was unloaded and the TEU was stored, the triatomines likely flew to and invaded the nearby residential areas. The rediscovery of this domestic vector of T. cruzi in Mexico warrants further investigation owing to the potential risk of transmission to the inhabitants of the study area.

Revisiting the hybridization processes in the Triatoma brasiliensis complex (Hemiptera, Triatominae): Interspecific genomic compatibility point to a possible recent diversification of the species grouped in this monophyletic complex.

Triatomines are hematophagous insects of great epidemiological importance, since they are vectors of the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. Triatoma brasiliensis complex is a monophyletic group formed by two subspecies and six species: T. b. brasiliensis, T. b. macromelasoma, T. bahiensis, T. juazeirensis, T. lenti, T. melanica, T. petrocchiae and T. sherlocki. The specific status of several species grouped in the T. brasiliensis complex was confirmed from experimental crossing and analysis of reproductive barriers. Thus, we perform interspecific experimental crosses between T. lenti and other species and subspecies of the T. brasiliensis complex and perform morphological analysis of the gonads and cytogenetic analysis in the homeologous chromosomes of the hybrids of first generation (F1). Besides that, we rescue all the literature data associated with the study of reproductive barriers in this monophyletic complex of species and subspecies. For all crosses performed between T. b. brasiliensis, T. b. macromelasoma, T. juazeirensis and T. melanica with T. lenti, interspecific copulas occurred (showing absence of mechanical isolation), hybrids were obtained, none of the male hybrids presented the phenomenon of gonadal dysgenesis and 100% pairing between the chromosomes homeologous of the hybrids was observed. Thus, we demonstrate that there are no pre-zygotic reproductive barriers installed between T. lenti and the species and subspecies of the T. brasiliensis complex. In addition, we demonstrate that the hybrids obtained between these crosses have high genomic compatibility and the absence of gonadal dysgenesis. These results point to reproductive compatibility between T. lenti and species and subspecies of the T. brasiliensis complex (confirming its inclusion in the complex) and lead us to suggest a possible recent diversification of the taxa of this monophyletic group.

Triatoma sordida (Hemiptera, Triatominae) from La Paz, Bolivia: an incipient species or an intraspecific chromosomal polymorphism?

BACKGROUND: Triatoma sordida is one of the main Chagas disease vectors in Brazil. In addition to Brazil, this species has already been reported in Bolivia, Argentina, Paraguay, and Uruguay. It is hypothesized that the insects currently identified as T. sordida are a species subcomplex formed by three cytotypes (T. sordida sensu stricto [s.s.], T. sordida La Paz, and T. sordida Argentina). With the recent description of T. rosai from the Argentinean specimens, it became necessary to assess the taxonomic status of T. sordida from La Paz, Bolivia, since it was suggested that it may represent a new species, which has taxonomic, evolutionary, and epidemiological implications. Based on the above, we carried out molecular and experimental crossover studies to assess the specific status of T. sordida La Paz. METHODS: To evaluate the pre- and postzygotic barriers between T. sordida La Paz and T. sordida s.s., experimental crosses and intercrosses between F1 hybrids and between F2 hybrids were conducted. In addition, cytogenetic analyses of the F1 and F2 hybrids were applied with an emphasis on the degree of pairing between the homeologous chromosomes, and morphological analyses of the male gonads were performed to evaluate the presence of gonadal dysgenesis. Lastly, the genetic distance between T. sordida La Paz and T. sordida s.s. was calculated for the CYTB, ND1, and ITS1 genes. RESULTS: Regardless of the gene used, T. sordida La Paz showed low genetic distance compared to T. sordida s.s. (below 2%). Experimental crosses resulted in offspring for both directions, demonstrating that there are no prezygotic barriers installed between these allopatric populations. Furthermore, postzygotic barriers were not observed either (since the F1 × F1 and F2 × F2 intercrosses resulted in viable offspring). Morphological and cytogenetic analyses of the male gonads of the F1 and F2 offspring demonstrated that the testes were not atrophied and did not show chromosome pairing errors. CONCLUSION: Based on the low genetic distance (which configures intraspecific variation), associated with the absence of prezygotic and postzygotic reproductive barriers, we confirm that T. sordida La Paz represents only a chromosomal polymorphism of T. sordida s.s.

Diversity and interactions among triatomine bugs, their blood feeding sources, gut microbiota and Trypanosoma cruzi in the Sierra Nevada de Santa Marta in Colombia.

Chagas disease remains a major neglected disease in Colombia. We aimed to characterize Trypanosoma cruzi transmission networks in the Sierra Nevada de Santa Marta (SNSM) region, to shed light on disease ecology and help optimize control strategies. Triatomines were collected in rural communities and analyzed for blood feeding sources, parasite diversity and gut microbiota composition through a metagenomic and deep sequencing approach. Triatoma dimidiata predominated, followed by Rhodnius prolixus, Triatoma maculata, Rhodnius pallescens, Panstrongylus geniculatus and Eratyrus cuspidatus. Twenty-two species were identified as blood sources, resulting in an integrated transmission network with extensive connectivity among sylvatic and domestic host species. Only TcI parasites were detected, predominantly from TcIb but TcIa was also reported. The close relatedness of T. cruzi strains further supported the lack of separate transmission cycles according to habitats or triatomine species. Triatomine microbiota varied according to species, developmental stage and T. cruzi infection. Bacterial families correlated with the presence/absence of T. cruzi were identified. In conclusion, we identified a domestic transmission cycle encompassing multiple vector species and tightly connected with sylvatic hosts in the SNSM region, rather than an isolated domestic transmission cycle. Therefore, integrated interventions targeting all vector species and their contact with humans should be considered.

Trends in taxonomy of Triatomini (Hemiptera, Reduviidae, Triatominae): reproductive compatibility reinforces the synonymization of Meccus Stål, 1859 with Triatoma Laporte, 1832.

BACKGROUND: Meccus' taxonomy has been quite complex since the first species of this genus was described by Burmeister in 1835 as Conorhinus phyllosoma. In 1859 the species was transferred to the genus Meccus and in 1930 to Triatoma. However, in the twentieth century, the Meccus genus was revalidated (alteration corroborated by molecular studies) and, in the twenty-first century, through a comprehensive study including more sophisticated phylogenetic reconstruction methods, Meccus was again synonymous with Triatoma. Events of natural hybridization with production of fertile offspring have already been reported among sympatric species of the T. phyllosoma subcomplex, and experimental crosses demonstrated reproductive viability among practically all species of the T. phyllosoma subcomplex that were considered as belonging to the genus Meccus, as well as between these species and species of Triatoma. Based on the above, we carried out experimental crosses between T. longipennis (considered M. longipennis in some literature) and T. mopan (always considered as belonging to Triatoma) to evaluate the reproductive compatibility between species of the T. phyllosoma complex. In addition, we have grouped our results with information from the literature regarding crosses between species that were grouped in the genus Meccus with Triatoma, in order to discuss the importance of experimental crosses to confirm the generic reorganization of species. RESULTS: The crosses between T. mopan female and T. longipennis male resulted in viable offspring. The hatching of hybrids, even if only in one direction and/or at low frequency, demonstrates reproductive compatibility and homeology between the genomes of the parents. CONCLUSION: Considering that intergeneric crosses usually do not result in viable offspring in Triatominae, the reproductive compatibility observed between the T. phyllosoma subcomplex species considered in the Meccus genus with species of the Triatoma genus shows that there is "intergeneric" genomic compatibility, which corroborates the generic reorganization of Meccus in Triatoma.

Chagas Disease Vectors of Paraguay: Entomoepidemiological Aspects of Triatoma sordida (Stål, 1859) and Development of an Identification Key for Paraguayan Triatomines Based on Cytogenetics Data.

Approximately 150,000 people are living with Chagas disease in Paraguay. Although the country has been since 2008 considered as one of the countries that succeeded in interrupted the vector transmission of Chagas by Triatoma infestans in houses of the eastern region, there are nine other species notified in the country that are potential vectors and also deserve attention from vector control programs. Thus, we carried out an entomoepidemiological study of T. sordida in the eastern and western regions of the country and we developed an identification key for Paraguay's triatomines based on cytogenetic data. Between the years 2003 to 2004, 271 specimens of T. sordida were captured in domestic, peridomestic, and wild ecotopes, with 131 insects caught in the eastern (Alto Paraguay, Boquerón and Pte. Hayes) and 140 in the western region of Paraguay (Guairá and Paraguarí). High rates of peridomicillary infestation were observed for both regions. Besides that, the natural infection of the captured insects was detected by optical microscopy in 12% and 10%, and by PCR in 21% and 20% in the eastern and western regions, respectively. Based on cytogenetic data from nine of ten species notified in Paraguay, an identification key was developed to differentiate all taxa. Thus, given the vectorial importance of T. sordida, we highlight the need for continued attention from Paraguay's vector control programs for this species. Further, we provide a taxonomic key that assists in the correct classification of Paraguayan triatomines.

Triatomine and Trypanosoma cruzi discrete typing units distribution in a semi-arid area of northeastern Brazil.

The occurrence of triatomine species, their bloodmeal sources and the discrete typing units (DTUs) of Trypanosoma cruzi isolated from them were determined in different municipalities of the state of Rio Grande do Norte, Brazil. Triatomine captures were carried out in the rural areas of 23 municipalities. The genotyping of T. cruzi isolates was performed using the mitochondrial cytochrome c oxidase subunit 2 (coii) gene, the D7 region of the 24Sα rDNA, and the spliced leader intergenic region (SL-IR). Five triatomine species were captured, and the most frequent was Triatoma brasiliensis (84.3%; 916/1086), which was found in 16 of the 23 municipalities surveyed, and infested all types of environment investigated. The TcI DTU was found in all mesoregions surveyed in 51.5% (17/33) of the culture-positive samples. In contrast, TcII (9.1%; 3/33) was detected in the Central mesoregion, while TcIII (27.3%; 9/33) was found in all mesoregions. The geographic distribution and spatial overlap of different DTUs was inferred using the superposition of the radius of occurrence of isolates and using ecological niche distribution modelling. Triatoma brasiliensis was found infected in all mesoregions and with all three T. cruzi DTUs, including mixed infections. With regard to bloodmeal sources, the DNA of rodents was found in triatomines infected with either TcI or TcIII, while that of domestic animals and humans was associated with both single and mixed infections. Our findings demonstrate that different DTUs of T. cruzi are widely dispersed among triatomines in our study area. The association of T. brasiliensis with several different mammalian hosts, as well as overlapping areas with different DTUs, suggests that this triatomine species may have an important role as a vector in both anthropic and sylvatic environments.

Population structure and genetic diversity of Triatoma longipennis (Usinger, 1939) (Heteroptera: Reduviidae: Triatominae) in Mexico.

Triatoma longipennis (Usinger) is an important vector of Trypanosoma cruzi in western, central and northern Mexico, due to its wide distribution, high infection rates, and epidemiological indices. However, its population genetics has not been completely characterized. In this study, the intra-specific relationships between different T. longipennis populations were analyzed from seven states in Mexico using mitochondrial cyt B as a marker. Our results show that the population of Jalisco was the most diverse, with the highest genetic and haplotypic variation (Hd = 0.978, π = 0.099 and θ = 0.079), even 25 times higher than some other populations analyzed. Heterogeneous migration and gene flow were observed without relation to their geographical distribution, that is, nearby populations may present high values of gene flow with low migration. In contrast, remote populations have low gene flow values with high migration. Genetic isolation was apparently present in the Guanajuato population, however, Mantel's analyzes to determine when an isolation by distance is present did not show correlation between genetic (FST) and geographic (Km) distances (P = 0.064). The STRUCTURE analyzes showed that populations such as Chihuahua, Jalisco, Nayarit and Michoacán appear to show a similar population structure, suggesting a common ancestor. Our results suggest two routes of diversification of T. longipennis highly influenced by anthropogenic effects. Elucidation of the population genetic structure of T. longipennis will help to better understand the role of gene flow and migration in the dispersal of this important Chagas disease vector.

Quantitative imagery analysis of spot patterns for the three-haplogroup classification of Triatoma dimidiata (Latreille, 1811) (Hemiptera: Reduviidae), an important vector of Chagas disease.

BACKGROUND: Spots and coloring patterns evaluated quantitatively can be used to discriminate and identify possible cryptic species. Species included in the Triatoma dimidiata (Reduviidae: Triatominae) complex are major disease vectors of Chagas disease. Phylogenetic studies have defined three haplogroups for Mexico and part of Central America. We report here our evaluation of the possibility of correctly discriminating these three T. dimidiata haplogroups using the pattern of the dorsal spots. METHODS: Digital images of the dorsal region of individuals from the three haplogroups were used. Image processing was used to extract primary and secondary variables characterizing the dorsal spot pattern. Statistical analysis of the variables included descriptive statistics, non-parametric Kruskal-Wallis tests, discriminant function analysis (DFA) and a neural classification network. RESULTS: A distinctive spot pattern was found for each haplogroup. The most differentiated pattern was presented by haplogroup 2, which was characterized by its notably larger central spots. Haplogroups 1 and 3 were more similar to each other, but there were consistent differences in the shape and orientation of the spots. Significant differences were found among haplogroups in almost all of the variables analyzed, with the largest differences seen for relative spot area, mean relative area of central spots, central spots Feret diameter and lateral spots Feret diameter and aspect ratio. Both the DFA and the neural network had correct discrimination values of > 90%. CONCLUSIONS: Based on the results of this analysis, we conclude that the spot pattern can be reliably used to discriminate among the three haplogroups of T. dimidiata in Mexico, and possibly among triatomine species.

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.

Revisiting the genetic variability of Brazilian peridomestic populations of the Chagas disease vector Triatoma sordida () (Hemiptera, Triatominae).

Triatoma sordida is an endemic species to South America, currently considered the species most frequently found in the peridomestic environment in Brazil. This triatomine has a wide ecological tolerability that allows it to inhabit several ecotopes and use different food sources. Although the species is considered predominantly peridomestic, it is also capable of colonizing households and forming numerous intra-household colonies, leading to its inclusion among the priorities for triatomine control campaigns in Brazil. All Brazilian populations of T. sordida are considered as T. sordida sensu stricto by chromosomal analyses (which highlights their epidemiological importance), although molecular studies that characterize the genetic diversity of these populations are scarce, being restricted only to the region of Minas Gerais. Thus, several populations of this vector distributed in the states of Bahia, Goiás, Mato Grosso do Sul, and Minas Gerais were analyzed using mitochondrial markers (cyt b and nd1). Low nucleotide diversity, high haplotypic diversity, low genetic distance, and high FST value were observed, as well as the formation of a monophyletic clade of the Brazilian populations of T. sordida, which confirms that this species has low genetic variability, with all specimens in Brazil grouped in T. sordida sensu stricto. In addition to the genetic and evolutionary importance for the knowledge of the biology of these vectors, these results are important from an epidemiological point of view, thus being able to direct vector control programs.

Phylogenetic and phenotypic relationships of the Triatoma sordida subcomplex (Hemiptera: Reduviidae: Triatominae).

Chagas disease, whose etiological agent is the protozoan Trypanosoma cruzi, affects millions of people worldwide and it is mainly transmitted by infected triatomine feces. Triatoma is the most diverse genus and one of the most important from an epidemiological point of view. Species of this genus are grouped into eight complexes and nine subcomplexes. Triatoma sordida subcomplex consists of T. garciabesi, T. guasayana, T. patagonica, and, T. sordida. Given the recent discussions on their phylogenetic status, this study aims to evaluate morphometric and genetics characters that group and distinguish T. garciabesi, T. guasayana, T. patagonica, and T. sordida, as well as to infer the phylogenetic relationships within the group and evaluate the proximity with T. rubrovaria subcomplex. The results corroborate the phylogenetic relationship of T. guasayana and T. patagonica with the T. rubrovaria subcomplex. Molecular data confirm the proximity of T. jurbergi, T. matogrossensis, T. vandae as T. garciabesi, and T. sordida. Together, genetic variability was shown between T. sordida populations.

Domestic Triatoma sanguisuga-Human Exposure in the South Carolina Coastal Region.

A collaborative investigation was initiated in rural coastal South Carolina in response to a reported triatomine bite. The eastern conenose bug, Triatoma sanguisuga, was identified and tested for Trypanosoma cruzi. The insect was negative by PCR, and no additional triatomines were found in the vicinity of the home. This is the first published report of a bite from T. sanguisuga in South Carolina despite the fact that triatomine vectors have been documented in the state since the 1850s, and specimens have been collected from homes in the past. Sylvatic T. cruzi reservoirs are common throughout the southeastern United States, and this case brings to light the possibility of human contact with infected triatomines in the state of South Carolina for public health and clinical and entomology professionals.

Identifying Chagas disease vectors using elliptic Fourier descriptors of body contour: a case for the cryptic dimidiata complex.

BACKGROUND: Triatoma dimidiata (Reduviidae: Triatominae) is an important vector of Chagas disease in various countries in the Americas. Phylogenetic studies have defined three lineages in Mexico and part of Central America. While there is a marked genetic differentiation, methods for identifying them using morphometric analyses with landmarks have not yet been fully resolutive. Elliptical Fourier descriptors (EFDs), which mathematically describe the shape of any closed two-dimensional contours, could be a potentially useful alternative method. Our objective was to validate the use of EFDs for the identification of three lineages of this species complex. METHOD: A total of 84 dorsal view images of individuals of the three lineages were used. Body contours were described with EFDs using between 5 and 30 harmonics. The number of obtained coefficients was reduced by a principal components analysis and the first axis scores were used as shape variables. A linear discriminant function analysis and an ordination plot of the discriminant analysis were performed using the shape variables. A confusion matrix of the ordination plot of the discriminant analysis was obtained to estimate the classification errors, the first five PC scores were statistically compared, and a neural network were then performed using the shape variables. RESULTS: The first principal component explained 50% of the variability, regardless the number of harmonics used. The results of discriminant analysis get improved by increasing the number of harmonics and components considered. With 25 harmonics and 30 components, the identification of haplogroups was achieved with an overall efficiency greater than 97%. The ordering diagram showed the correct discrimination of haplogroups, with only one error of discrimination corroborated by the confusion matrix. When comparing the first five PC scores, significant differences were found among at least two haplogroups. The 30 multilayer perceptron neural networks were also efficient in identification, reaching 91% efficiency with the validation data. CONCLUSIONS: The use of EFD is a simple and useful method for the identification of the main lineages of Triatoma dimidiata, with high values of correct identification.

Molecular data supports monophyly of Triatoma dispar complex within genus Triatoma.

The genus Triatoma contains numerous species, principal or secondary vectors of Chagas disease, which have been included in the three main lineages of Triatomini tribe based on morphological and biogeographical characteristics: North American, South American, and T. dispar complex. The three members of the T. dispar complex are distributed in Ecuador. This complex has been scarcely studied through molecular approaches, and the taxonomic position of this complex is not confirmed. In this study, we explored the phylogenetic relationships within the genus Triatoma, including five species from North and Central America, six from South America, and the three species belonging to the T. dispar complex. Partial sequences of four mitochondrial genes (Cyt b, COII, 16S-rRNA, 12S-rRNA) and two nuclear genes (18S-rRNA, ITS2) were obtained from 74 specimens. Phylogenetic trees were built with concatenated and single sequences through maximum likelihood (ML), maximum parsimony (MP), and Bayesian methods. The trees built using concatenated sequences showed three main branches (clusters) highly supported by significant bootstrap values; the T. dispar complex appeared as a monophyletic group separate from species of North and Central American origin and South American origin. On the contrary, for each gene tree, the three main clusters were not always significantly supported, mostly because genetic information is dramatically reduced when a single gene is considered. Consequently, concatenation of genes gives relevant results and is highly recommended for further in-depth examination of the relationships of several species and complexes of triatomines that remain unresolved. Moreover, our current molecular data fully revealed the division of genus Triatoma into at least three main genetic groups.

Triatoma brasiliensis species complex: characterization of the external female genitalia.

The Triatoma brasiliensis complex is composed of six species (Triatoma bahiensis, T. juazerensis, T. lenti, T. melanica, T. petrocchiae, and T. sherlocki) and two subspecies (T. brasiliensis brasiliensis and T. b. macromelasoma). Phylogenetic studies have shown that it is a monophyletic group, but the morphological relationships among the members can be better understood. We applied a tool of high resolution based on morphology to identify details that characterize the female genitals for each member of this group. The external genital structures of five to fifteen females of each taxon were analyzed. Insects had their abdomen transversely cut to separate the last abdominal segments, and representative illustrations obtained via scanning electron microscopy (SEM) are shown. Observations were performed in the dorsal and the ventral views. The morphological characterization of female external genitalia allowed a differentiation of each species/subspecies. The subspecies T. b. brasiliensis and T. b. macromelasoma were the most similar. On the other hand, dorsal view of structures evidenced some unique characteristics for T. petrocchiae and T. sherlocki. Regarding these structures for T. petrocchiae, the differentiation was in accordance with molecular studies, since this species exhibits the highest genetic distances in relation to other species of T. brasiliensis complex.

Distribution, genetic characteristics and public health implications of Triatoma rubrofasciata, the vector of Chagas disease in Guangxi, China.

BACKGROUND: Triatomines are natural vectors of Chagas disease and are mainly prevalent in the Americas. In China, previous data from decades ago showed that there were two species of triatomine bugs, Triatoma rubrofasciata and T. sinica. However, the distribution, genetic characteristics and public health implications of triatomines in China are still relatively unknown. In order to gain knowledge on the distribution, genetic characteristics and public health implications of the triatomines in Guangxi, China, an entomological-epidemiological study and genetic research was conducted. METHODS: Different methods were used to elucidate the distribution of triatomines in Guangxi including consultations with county-level Center for Disease Prevention and Control staff and village doctors, the distribution of educational material on triatomines though the internet and social media apps such as Wechat and QQ, and conducting manual inspections and light trapping to collect triatomines. The morphological characteristics of the collected triatomines were identified under light microscopy. The mitochondrial 16S rRNA, cytochrome b (cytb) genes and nuclear 28S rRNA gene were amplified, sequenced and used in phylogenetic analyses. RESULTS: A total of 305 triatomines were captured from 54 different sites in 13 cities in Guangxi. All collected bugs were identified as T. rubrofasciata based on morphology. Most triatomine collection sites were around or inside houses. Four triatomines bite cases were observed during the investigation indicating that triatomine bites are common, the bites can cause serious anaphylaxis and skin papules and urticaria, suggesting a systemic skin response. The 16S rRNA, 28S rRNA and cytb sequence analyses of T. rubrofasciata from Guangxi and other countries showed that T. rubrofasciata sequences from different regions exhibit a high similarity, with no geographical differences. The phylogenetic tree based on the 16S rRNA and cytb genes showed that T. rubrofasciata sequences from different regions and continents were in the same cluster, indicating no differentiation among different geographical populations. CONCLUSIONS: Our study showed that T. rubrofasciata is widely distributed in Guangxi and that people are commonly bitten by this insect in some regions. This highlights the need to enhance surveillance for and control of T. rubrofasciata and to strengthen the monitoring of imported Trypanosoma cruzi in China. The 16S rRNA, 28S rRNA and cytb sequence analyses of T. rubrofasciata from different regions and continents suggested that T. rubrofasciata populations exhibit high similarity, and the clustering in the phylogenetic analyses indicates that T. rubrofasciata has a close ancestor originating in the Americas.

Molecular cytotaxonomy of the Triatoma brasiliensis species subcomplex (Hemiptera, Triatominae).

The Triatoma genus is paraphyletic, and its species are grouped into complexes and subcomplexes. Given the fact that species that make up a given subcomplex generally share chromosomal traits, we analyzed the distribution of AT- and CG-rich DNA of the T. brasiliensis species subcomplex, in order to establish affinities among members of the T. brasiliensis subcomplex based on chromatin and chromosome traits and develop an identification key for the four monophyletic Triatoma subcomplexes from South America. All species exhibited a CG-rich X sex chromosome and autosomes, as well as an AT-rich Y sex chromosome. This feature can be used as a diagnostic characteristic to determine whether a given species is a member of the T. brasiliensis subcomplex, because it enables the differentiation of these species from all Triatoma of South America. Thus, we confirmed the chromosomal relationship of the T. brasiliensis species subcomplex and developed a dichotomous key based on the chromocenter to differentiate the species from this subcomplex from the other monophyletic Triatoma subcomplexes from South America.

Prezygotic isolation confirms the exclusion of Triatoma melanocephala, T. vitticeps and T. tibiamaculata of the T. brasiliensis subcomplex (Hemiptera, Triatominae).

Chagas disease is caused by the Trypanosoma cruzi and transmitted mainly by triatomines. Triatoma is a paraphyletic group and the species of this genus are grouped into complexes and subcomplexes. Morphological data and geographical distribution grouped initially T. melanocephala, T. vitticeps and T. tibiamaculata in the T. brasiliensis subcomplex. However, karyotypic and phylogenetic analysis suggested the exclusion of T. melanocephala, T. vitticeps and T. tibiamaculata from this subcomplex. Considering that studies of experimental crosses can help to understand the systematics of species, we performed experimental crosses between T. melanocephala, T. vitticeps and T. tibiamaculata with T. b. brasiliensis. No crosses resulted in hybrids. Taking into account that the species of the T. brasiliensis subcomplex do not present interspecific prezygotic barriers, the characterization of reproductive barriers shows that T. melanocephala, T. vitticeps and T. tibiamaculata do not present an evolutionary proximity relationship with the species of this subcomplex. Thus, we confirmed the exclusion of these species from the T. brasiliensis subcomplex and we emphasize the importance of experimental crosses for evolutionary studies.

Biological attributes of the kissing bug Triatoma rubrofasciata from Vietnam.

BACKGROUND: Triatoma rubrofasciata is the only kissing bug species distributed globally. In the Americas, this species transmits the parasite Trypanosoma cruzi, responsible for Chagas disease. The presence of T. rubrofasciata in several Asian countries has greatly increased recently. In Vietnam, it is found in large numbers, closely associated with human environments. Although T. rubrofasciata from Asia is not infected with Tryp. cruzi, it carries other parasites such as Trypanosoma lewisi and Trypanosoma conorhini. Reports of bites by T. rubrofasciata have increased significantly in several places of Vietnam, becoming a public health problem as it produces severe anaphylactic reactions. METHODS: Specimens of T. rubrofasciata were collected from seven provinces in central Vietnam. We analyzed different biological attributes (life-cycle, starvation resistance, feeding and reproductive capacities) and genetic characteristics (chromosomes and DNA sequences) of T. rubrofasciata from Vietnam and compared them with Brazilian specimens. Natural infection with Tryp. conorhini and Tryp. lewisi were analyzed in a sample of 100 collected insects. RESULTS: Species identification of T. rubrofasciata from central Vietnam was corroborated by genetic markers. Cytogenetic analyses showed that T. rubrofasciata from central Vietnam share the same chromosomal characteristics with individuals from Brazil and Hanoi. DNA sequence analyses of a mitochondrial cytochrome b gene fragment showed little variation between Old and New World specimens. Our study sample, compared with Brazilian individuals, showed a higher survival capacity revealed by a higher hatching rate (98% compared with 80.5%), a larger amount of blood taken in single meal and long-term starvation resistance. Furthermore, this species had a high natural rate of infection with Tryp. conorhini (46%) and Tryp. lewisi (27%). CONCLUSIONS: For T. rubrofasciata of Vietnam, a high rate of fecundity throughout the year, a high capacity for starvation, and its occurrence in synanthropic environments of urban areas with a high availability of food sources are risk factors to be taken into account by vector control campaigns. The several allergic reactions caused by their bites and their high infection with Tryp. lewisi highlight the need to implement specific control programmes for T. rubrofasciata in Vietnam.