Characterization of Triatoma infestans (Klug, 1834) (Hemiptera: Heteroptera, Reduviidae) from Residual Foci in the States of Bahia and Rio Grande do Sul, Brazil, Using Geometric Morphology.

Triatoma infestans is one of the main vectors of Chagas disease in Latin America. Although the species is under control in most Latin countries, it is still necessary to maintain epidemiological surveillance. The present study aims to characterize T. infestans populations from residual foci in Bahia and Rio Grande do Sul, Brazil, comparing them with natural populations in Argentina and Bolivia. For this, we adopt the geometric morphometry of the heads. It is possible to report the morphometric variety of the studied populations. In addition, we show that the size of the heads contributes to the differentiation between populations, while the shape has less power to discriminate groups. Furthermore, we show that some natural populations have morphometric proximity to residual populations, suggesting a relationship between these triatomines. Our data do not support the origin of residual populations but demonstrate the importance of new studies with other techniques to understand the dynamics of distribution and reintroduction of these vectors in Brazilian territory.

Integrative taxonomy and phylogenetic systematics of the Triatoma rubrovaria subcomplex (Hemiptera, Triatominae).

Triatoma rubrovaria subcomplex consists of T. carcavalloi, T. circummaculata, T. klugi, T. limai, T. oliveirai, T. pintodiasi, T. rubrovaria, T. patagonica and T. guasayana, which can be vectors of Trypanosoma cruzi, the etiologic agent of Chagas disease. In this study, morphological, morphometric, and genetic characters of T. circummaculata, T. pintodiasi, T. carcavalloi, T. klugi, and T. rubrovaria were analyzed in view of the integrative taxonomy and phylogeny of the T. rubrovaria subcomplex. Molecular studies were carried out through the sequencing and analysis of the mitochondrial genes COI and CytB, nuclear genes ITS I, ITS 2, 16S, and 28S from rDNA and rescued a monophyletic group. Furthermore, differential morphological characters were found among the five species in the pronotum, scutellum, stridulatory sulcus, male genitalia, and external female genitalia. Finally, morphometric analyses made it possible to differentiate the five species. Phylogenetic analyzes rescued the relationship of T. pintodiasi with members of the T. rubrovaria subcomplex and demonstrated that this subcomplex is a monophyletic group composed of the species T. carcavalloi, T. circummaculata, T. klugi, T. guasayana, T. limai, T. oliveirai, T. patagonica, T. pintodiasi, and T. rubrovaria. Furthermore, through integrative taxonomy, it was possible to confirm the specific status of the species T. carcavalloi, T. circummaculata, T. pintodiasi, T. klugi, and T. rubrovaria, offering new useful morphological characters for the differentiation and characterization of these potential vectors and distributed in Southern Brazil.

Growth Curve, Morphological and Molecular Characterization of Two Strains of Trypanosoma cruzi (Kinetoplastida, Trypanosomatidae) isolated from Triatoma sherlocki (Hemiptera, Reduviidae, Triatominae).

BACKGROUND: Trypanosoma cruzi presents great variability in morphology, virulence, pathogenicity, avoidance of the host immune system, and antigenic constitution, associated with different clinical manifestations of the disease. METHODS: Two strains of T. cruzi were cultivated in liver infusion tryptose to determine growth kinetics, morphometry and molecular characterization using restriction fragment length polymorphism polymerase chain reaction. RESULTS: The biological parameters showed sharp growth by the 7th day. Morphologically, both strains showed short and thin forms and were classified as Group I. CONCLUSION: Group TcI presents cardiac manifestations and T. sherlocki is adapting to the home environment, requiring attention to future problems.

Immature instars of three species of Rhodnius Stål, 1859 (Hemiptera, Reduviidae, Triatominae): morphology, morphometry, and taxonomic implications.

BACKGROUND: Among the 18 genera of the Triatominae subfamily, three stand out for their diversity and epidemiological importance: Triatoma, Panstrongylus, and Rhodnius. Rhodnius includes 21 species that can transmit Trypanosoma cruzi (the etiological agent of Chagas disease, also known as American trypanosomiasis) and Trypanosoma rangeli. The Rhodnius prolixus complex comprises seven species, including Rhodnius marabaensis, Rhodnius prolixus, and Rhodnius robustus, which occur in the northern region of Brazil. Since both adults and immatures can carry T. cruzi, in this study the five nymphal instars of the three species mentioned were dorsally characterized. METHODS: Using microscopy, morphometrics, and geometric morphometrics, the present work measures and describes the morphological characters of the five nymphal instars of R. marabaensis, R. prolixus, and R. robustus. RESULTS: The study enabled the characterization of all five nymphal instars, as well as the distinction between the three species in each of their instars. CONCLUSIONS: The morphological, morphometrics of the head, thorax, and abdomen and geometric morphometrics studies of the head enabled the specific distinction of these three species in all five instars.

New in morphometry: Geometric morphometry of the external female genitalia of Triatominae (Hemiptera: Reduviidae).

The study of geometric morphometry has an impact on Triatominae studies. Currently, several taxonomic and systematic studies use this approach. The Triatominae subfamily comprises three fossil species and 154 extant species potentially capable of transmitting Trypanosoma cruzi, the causative agent of Chagas disease. This study aims to evaluate the external female genitalia of adult triatomines using multivariate geometric morphometric approaches, not only for validation but also for systematic inferences. Specimens belonging to the genera Panstrongylus, Psammolestes, Rhodnius, and Triatoma were evaluated, in addition to two species previously included in Triatoma: T. longipennis and T. phyllosoma. The results show that the external female genitalia have operational morphology and allow characterization of the species and the genera of the Triatominae. In addition, the multivariate technique enabled delimitation of the phylogenetic relationships of the subfamily, presenting results consistent with systematic studies. It can be concluded that the external female genitalia evaluated by geometric morphometry is a useful character for the taxonomy and systematics of Triatominae.

A review of the taxonomy and biology of Triatominae subspecies (Hemiptera: Reduviidae).

Due to its public health importance, as vectors of Chagas disease, the subfamily Triatominae, has received the attention of taxonomists. Knowing how to correctly identify these insects is of paramount importance for the surveillance and control of these vectors. Over many years, more than 40 triatomine subspecies were proposed. Infraspecific taxa are accepted as real entities in nature and recognized as biologically significant. Infraspecific diversity and population subdivision are frequently confused with the underlying mechanisms that lead to the formation of new species and species-level diversity. The distinction between infraspecific and interspecific divergence processes has received far less attention than species delimitation. In this work, new literature data on subspecies included in the subfamily Triatominae are presented. Data were compiled about Triatoma brasiliensis, Triatoma breyeri, Triatoma circummaculata, Triatoma dimidiata, Triatoma incrassata, Triatoma infestans, Triatoma phyllosoma, Triatoma protracta, Triatoma recurva, Triatoma rubida, Triatoma sanguisuga, Triatoma vitticeps, Panstrongylus megistus, Paratriatoma hirsuta, and Paratriatoma lecticularia subspecies. Although several of these subspecies have been synonymized, herein, we emphasize that some subspecies may be valid. Therefore, further studies are needed on the taxonomy, evolution, phylogeny, biogeography, ecology, physiology, and behavior of species to reinforce, or to invalidate the infraspecific status in the subfamily.

Checklist and phenetics studies of nymphs of two species of triatomines: Triatoma lenti Sherlock & Serafim, 1967 and Triatoma sherlocki Papa, Jurberg, Carcavallo, Cerqueira, Barata, 2002 (Hemiptera: Reduviidae: Triatominae).

INTRODUCTION: Triatoma lenti and Triatoma sherlocki are endemic species of the State of Bahia, located in northeastern Brazil, where they have records of domiciliation in the human environment. In view of the epidemiological aspect and taxonomic importance of these species for the systematics of the Triatoma genus, a study was carried out with nymphs of all five instars. METHODS: An extensive review of studies on nymphs from the subfamily Triatominae is presented. Morphology was studied using a scanning electron microscope and morphometric analyses. RESULTS: The morphological study allowed us to characterize and discriminate species by means of scanning electron microscope of the last abdominal segment. In addition, the results show morphometric variability, with the total size of the head that best discriminates the species. CONCLUSIONS: Studies on nymphs are fundamental to the ecosystem; however, the literature on the immature forms of certain groups is scarce, difficult to use, or nonexistent. Therefore, this study includes morphological and morphometric data of the nymphal instars of T. lenti and T. sherlocki, corroborating the specific taxonomy of these species.

Characterization of Female External Genitalia and Eggs of Four South American Species of the Triatoma Laporte, 1832 Genus (Hemiptera: Reduviidae: Triatominae).

Triatoma is the most diversified and one of the most important genera from an epidemiological perspective. Given the difficulty in identifying some species of the Triatoma genus, morphological, histological, and morphometric studies were performed to provide new characters that make it possible to differentiate T. garciabesi, T. guasayana, T. patagonica, and T. sordida sensu stricto, triatomines that overlap geographically and have vector potential. Through the external female genitalia, as well as morphology, morphometry, and histology of eggshells, it was possible to discriminate the four species. In addition, this study reinforces the taxonomic validity of T. garciabesi and provides new data for discussion on systematic issues of T. guasayana and T. patagonica.

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.

Biological Aspects of Rhodnius montenegrensis (Hemiptera, Reduviidae, Triatominae) Under Laboratory Conditions.

Triatominae are insects notorious as vectors of Trypanosoma cruzi, which is the etiologic agent of Chagas disease, and other trypanosomatids. Triatomines of the genus Rhodnius are primarily sylvatic, nevertheless the occurrence of native species that invade households suggests their possible role in the transmission of Chagas disease. Rhodnius montenegrensis was first described in 2012, but the biological aspects of this species are still unknown. This study aimed to analyze the biological aspects of R. montenegrensis under laboratory conditions. The emergence rate was 63.0%, the mean time required for the emergence was 13.9 ± 1.7 days, the biological cycle from egg to adult phase occurred in 105.2 ± 9.2 days, the number of bloodmeals required for each nymphal stage to reach the next stage varies between a minimum of two and a maximum of seven. The weight gained after a bloodmeal varied between 10.6 times on 1st-instar nymphs and 3.9 times on 5th-instar nymphs. The adult specimens had the lowest gain of weight, reaching 2.2 times on females and 1.6 times on males. The sex ratio observed was 1:1. These data are relevant to understand the life cycle of this new described species and to elaborate more effective vector control strategies.

First Report of Natural Infection with Trypanosoma cruzi in Rhodnius montenegrensis (Hemiptera, Reduviidae, Triatominae) in Western Amazon, Brazil.

Rhodnius montenegrensis was described in 2012. Since then, reports of the occurrence of this species associated with palm trees near households in Rondônia have been published. This study aimed to analyze the natural infection of R. montenegrensis with trypanosomatids in the municipality of Monte Negro, Rondônia, Brazilian western Amazon. Capture of triatomines occurred in Attalea speciosa (babassu) specimens around households. Twelve of the 72 captured triatomines were identified by morphological and morphometric characters, by molecular analysis made using the PCR, DNA sequencing, and phylogenetic analyses. The intestinal content was collected from 60 of these 72 specimens. The positivity for trypanosomatids was confirmed by examining the intestinal content followed by PCR amplification of the cathepsin L-like gene specific for Trypanosoma cruzi (PCR-DTcrCatL) and Trypanosoma rangeli (PCR-DTraCatL). Of the 60 specimens analyzed by microscopy, 22 (36.7%) were positive for trypanosomatids in the intestinal content analysis. Of these 22 specimens of R. montenegrensis, 16 (72.7%) were infected with T. cruzi, 2 (9.1%) were infected with T. rangeli, 2 (9.1%) had mixed infection with T. cruzi and T. rangeli, and 2 were negative (9.1%). These data suggest the need for attention of the health surveillance system of Chagas disease in the State of Rondônia, as this study points out to another potential vector of the disease.

Reproductive aspects of Chagas disease vectors (Hemiptera, Triatominae) with anatomical teratologies.

As Chagas disease has no cure, vector control is the most effective method of preventing this neglected disease. Generally the anatomical teratologies are associated with hormonal dysfunction of the corpus allatum (juvenile hormone), presents genetic basis or unfavorable conditions, and are recessive and not sex linked. Thus, we characterize the male gonads and spermatogenesis of triatomines with anatomical teratologies to evaluate if the malformations interfere in the gametogenesis of these vectors. All teratogenic triatomines analyzed did not present presented gonadal dysgenesis and alterations in spermatogenesis. Thus, we characterize the presence of anatomical teratologies in some species of triatomines and demonstrate that these malformations in the external morphology do not interfere in the gonads and gametogenesis of these vectors. In addition, although new studies with the corpus allatum should be conducted we suggesting that the use of juvenile hormones does not present effectiveness in the reproductive control of these vectors.

Third record of Rhodnius amazonicus and comparative study with R. pictipes (Hemiptera, Reduviidae, Triatominae).

Rhodnius amazonicus Almeida et al. (1973) is a triatomine of rare occurrence. This species was found for the third time in Breves city, Pará state, Brazil. Morphometric and morphological studies were carried out on one male and one female. Lent and Wygodzinsky (1979) considered this species as a synonym of R. pictipes, until its revalidation after 23 years by Bérenger and Pluot-Sigwalt (2002). Considering the synonym mentioned above, a comparative study between these two species was performed in order to report the third encounter of this species, and increase the number of morphological characters that distinguish R. amazonicus from R. pictipes. The dorsal side of the head, the dorsal and ventral portions of the thorax, the dorsal, ventral and posterior sides of the female genitalia, eggs and the median process of the pygophore were examined by scanning electronic microscopy (SEM). The head, thorax, abdomen and egg parameters of these two species were also measured. The identification of characters on the head, stridulatory sulcus, mesosternum, metasternum, scutellum, process of urotergite I, external female genitalia, pygophore and eggs made the distinction between R. amazonicus and R. pictipes possible. This study has shown the new characters recorded and described for the first time for R. amazonicus and R. pictipes by SEM here made it possible to confirm the morphological separation between these two species. The morphometric analysis also confirmed that the above two taxa are different at specific level.

Combined phylogenetic and morphometric information to delimit and unify the Triatoma brasiliensis species complex and the Brasiliensis subcomplex.

"Triatoma brasiliensis species complex" was defined as a monophyletic group of the species: T. brasiliensis, T. juazeirensis, T. melanica, and T. sherlocki. An alternative grouping scheme proposed the concept of "Brasiliensis subcomplex" which included the former species together with T. melanocephala, T. petrocchiae, T. lenti, T. tibiamaculata, and T. vitticeps. To evaluate the relationship among these taxa we combined the results obtained with four mitochondrial genes (12S, 16S, COI and Cytb, adding to 1811bp) and geometric morphometric analysis of wings and heads. Panstrongylus megistus was included in the analysis as it was previously found related to T. tibiamaculata, T. melanocephala and T. vitticeps. The results of both molecular and morphometric approaches clearly grouped the species analyzed into two monophyletic units, supported by both genetic and wing variability. The first one (G1) comprises the four species originally included in the T. brasiliensis species complex plus T. lenti and T. petrocchiae. The second group (G2) was composed by T. melanocephala, T. tibiamaculata and T. vitticeps, and remarkably, P. megistus if considering wing variability and phylogenetic results. Nevertheless, geometric morphometrics of heads provided a quantitative measurement that discriminates Panstrongylus from the Triatoma species based on the position of the antennal insertion relative to eyes, as it is used as the generic distinctive character. The discrepancy among approaches questions the validity of this character to define Panstrongylus genus. Independently of the chosen group definition -"T. brasiliensis species complex" or "Brasiliensis subcomplex"-we propose to delimit it to species of G1 that are all associated with the Caatinga biome in the Brazilian Northeast. G2 are the ones associated with the Atlantic Forest biome.