Comparative egg morphology of six Meccus species and Triatoma recurva (Stål, 1868) Hemiptera: Reduviidae

We investigated the morphology and morphometry of eggs from the colonies of the Entomology Laboratory at ENCB‐IPN belonging to six species of the genus Meccus and Triatoma recurva, using both light and scanning electron microscopy. Egg ornamentation is an important parameter to consider in the differentiation of species. Samples were observed under a scanning electron microscope. Measurements of fifty eggs per species included length, width, and opercular diameter, which were used for the morphometric analysis. The results showed that the seven species display a polygonal ornamentation dominated by hexagons; the operculum shows also a polygonal ornamental characteristic in each species. Morphometry provided valuable information for discriminating between closely related species of the genus Meccus and Triatoma recurva, a species akin to this group, thereby facilitating the complete discrimination of these species.     

External female genitalia of six species of the genus Meccus (Hemiptera: Reduviidae: Triatominae)

Triatomine classification is based on morphological characteristics. Studies have been conducted to improve their identification by observing many characteristics. However, there are problems of differentiating among highly interrelated species and new criteria are required. The purpose of this study was to determine the morphological differences in the external female genitalia of M. pallidipennis, M. longipennis, M. picturatus, M. bassolsae, M. mazzottii, and M. phyllosomus in order to distinguish among species using scanning electron microscopy. Observations were made of the dorsal, posterior, lateral, and ventral views of the female external genitalia for each species. In the six species we studied, relevant differences were observed in the dorsal view of the X segment, as well as the IX, VIII, and VII tergites. In the posterior and lateral view, the most visible differences were registered in the gonocoxite size of the segments VIII, IX, and X. Finally, in the ventral view of the VII sternite, differences among species were observed in the size of the inflection in the top and upper corner. Our results show that it was possible to differentiate among the triatominae species for each of the four views by using a scanning electron microscope to analyze morphological characteristics of the VII, VIII, IX, and X abdomen segments.     

Morphology and morphometry of the scutellum of six species in the genus Meccus (Hemiptera: Triatominae)

We studied the morphology and morphometry of scutella from six species of the hemipteran genus Meccus to identify new tools to help solve taxonomic problems in closely related insect species of epidemiological relevance. Scutellum samples were observed by scanning electron microscopy and were subjected to morphometric analysis. The results mainly show differences in central depression shape, posterior process, and vestiture. We found significant dimensional differences in scutellum morphometry and a clear sexual dimorphism among species. A combination of morphology and morphometry can be used to differentiate among species of the genus Meccus.     

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 revaluated 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.”     

Morphological study of eggs from five Mexican species and two morphotypes in the genus Triatoma (Laporte, 1832)

We describe and compare the morphology and morphometry of the egg exochorion for five species and two morphotypes of Mexican triatomines with scanning electron microscopy. The results show differences in egg ornamentation for each species, including between morphotypes. Polygonal ornamentation was observed in each species, including pentagons, octagons and, in certain cases, a majority of hexagons. We observed small perforations in T. protracta protracta, small spheres on the T. lecticularia polygons, a crown with festoons in T. barberi, and less complex ornamentation in T. mexicana. Through morphometric analysis, significant differences in egg dimensions were determined for the studied species.     

New structures for goat corrals to control peridomestic populations of Triatoma infestans (Hemiptera: Reduviidae) in the Gran Chaco of Argentina

Goat production is an important economic activity for rural communities in the Gran Chaco of Argentina. Goat corrals are important for the survival of peridomestic populations of Triatoma infestans. This study evaluated the impact of modifying the traditional structure of goat corrals on T. infestans populations and goat productivity in the region of Los Llanos (La Rioja). Thirty-nine experimental corrals were constructed and 57 traditional corrals were used as controls. We evaluated the infestations of the control and experimental corrals for five years following construction of the structures. The results showed that the new structures did not prevent the colonization, although it enhanced the detection of infestation at low densities of T. infestans. No significant difference was found in T. infestans population abundance between control and experimental corrals, probably because of the different detectability in the two types of structures, especially among the small nymphs. Although goat productivity average was higher in experimental than in control corrals, no significant difference was found because of high variability. The new structures can be used as a complement to promote the development of rural communities. Acceptability and adoption of the new corrals by the owners was high, as the enclosures offered better protection for the goats, increased growth of kids and facilitated herd handling.     

Distribution of triatomine (Hemiptera: Reduviidae) vectors of Chagas disease in the state of Hidalgo,Mexico

Chagas disease represents a significant public health problem in Mexico. In the state of Hidalgo, studies on the presence of triatomines are scarce and restricted to a few locations. To determine the risk of transmission in the state of Hidalgo and stratify the vector potential, the distribution of Triatominae was surveyed from 2015 to 2016 in collaboration with primary health care services and local communities. A total of 570 specimens was collected in 278 houses in 25 municipalities. The species of Triatominae detected were T. dimidiata with 391 samples, T. mexicana with 159 samples, T. gerstaeckeri with 17 samples, and T. barberi with three samples. The samples were collected in domestic and peridomestic areas. The general index of natural infection was 9.8%. Indices of colonization and crowding were determined for species and municipality and results were variable.     

Influence of blood meal source on the biological parameters of Triatoma pallidipennis (Heteroptera: Reduviidae) from Mexico

Chagas disease is one of the most important vector-borne diseases in Latin America, including Mexico. Triatoma pallidipennis (Stål) (Hemiptera: Reduviidae) is a Mexican triatomine vector commonly associated with different hosts. The influence of six blood meals (rabbits, rats, mice, dogs, cats and chickens) on six biological parameters of the biology of T. pallidipennis was evaluated. A significant difference was found in the period of egg-to-adult development between the five mammalian feeds (mean 195 days) and the chicken feed (221 days). The probability of survival was significantly lower in the chicken cohort (0.285). The total number of blood meals to moult from the first instar to the adult stage was the highest in the chicken cohort (10–15). This cohort had the significantly highest rate of females at the end cycle. The mean number of eggs laid per female and the egg eclosion rate were similar among the six food sources. Most results seemed to be influenced by the higher nutritional quality of the mammalian blood compared to the bird's blood and the increased energy expenditure required for the digestion of bird blood. These results clearly show that T. pallidipennis, unlike other triatomine species, has a high reproductive capacity when feeding on different hosts.     

Phylogenetic incongruence inferred with two mitochondrial genes in Mepraia spp. and Triatoma eratyrusiformis(Hemiptera,Reduviidae)

Mitochondrial DNA (mtDNA) is widely used to clarify phylogenetic relationships among and within species, and to determine population structure. Due to the linked nature of mtDNA genes it is expected that different genes will show similar results. Phylogenetic incongruence using mtDNA genes may result from processes such as heteroplasmy, nuclear integration of mitochondrial genes, polymerase errors, contamination, and recombination. In this study we used sequences from two mitochondrial genes (cytochrome b and cytochrome oxidase subunit I) from the wild vectors of Chagas disease, Triatoma eratyrusiformis and Mepraia species to test for topological congruence. The results showed some cases of phylogenetic incongruence due to misplacement of four haplotypes of four individuals. We discuss the possible causes of such incongruence and suggest that the explanation is an intra-individual variation likely due to heteroplasmy. This phenomenon is an independent evidence of common ancestry between these taxa.     

Morphological and histological study of eggs of six species of the Triatoma genus (Hemiptera: Reduviidae)

The objective of this study was to analyze the morphological and histological characteristics of eggs from six triatomine species in order to obtain a generic and specific characterization of this group of Trypanosoma cruzi vectors and better understand their phylogenetic and taxonomic aspects. The eggs of the respective species came from the collection of the Laboratório de Triatomíneos e Culicídeos da Faculdade de Saúde Pública/USP, from the Insetário do Servi?o Especial de Saúde de Araraquara - SESA and from the Triatomine insectary of the Laboratório Nacional e Internacional de Referência em Taxonomia de Triatomíneos (FIOCRUZ), in Rio de Janeiro. Morphological studies were done with optical microscopy and scanning electron microscopy. Histological analyses used only optical microscopy. All of the species analyzed showed a predominance of hexagonal cells. The exchorion of Triatoma breyeri Del Ponte, T. costalimai Verano Galv?o, and T. tibiamaculata (Pinto) consists of unornamented cells, overlapping or slightly separated, with a smooth, padded appearance. Meanwhile, in T. matogrossensis Leite Barbosa, T. sherlocki Papa, Juberg, Carcavallo, Cerqueira Barata, and T. williami Galv?o, Souza Lima, the exchorion cells are ornamented on their entire surface with perforations and fissures. Egg histology showed undulations, pores, and orifices. The study helps to expand the generic and specific knowledge of the Triatominae subfamily. The characteristics identified in the exchorion of the eggshells can help separate close species. These new parameters will back the elaboration of future dichotomous keys, help to determine each vector species role and expand knowledge of the various species.     

Comparison of biological fitness in crosses between subspecies of Meccus phyllosomus (Hemiptera: Reduviidae: Triatominae) in southern Mexico

Understanding the biological parameters of some triatomine subspecies of Meccus phyllosomus (Burmeister) is a crucial first step in estimating the epidemiologic importance of this group. Biological parameters related to hatching, lifetime, number of blood meals to molt, percentage of females at the end of the cycle, number of laid eggs, and mortality for each instar of 3 M. phyllosomus subspecies [M. p. mazzottii (Usinger), M. p. pallidipennis (Stål), and M. p. phyllosomus] and their laboratory hybrids were evaluated and compared. No significant differences (P > 0.05) were found among the experimental hybrids (MaPa, MaPhy, PaPhy) and reciprocal cohorts. In 5 (hatching, number of blood meals to molt, accumulative mortality, percentage of females, and mean number of laid eggs) of the 6 studied parameters (with the exception of development time), the hybrid cohorts had better fitness results than the parental cohorts involved in each set of crosses. The increase in hybrid fitness found in our study could lead to an increase in the epidemiologic risks caused by transmission of Trypanosoma cruzi to humans.     

Abundance and nightly activity behavior of a sylvan population of Triatoma dimidiata (Hemiptera: Reduviidae: Triatominae) from the Yucatan,México

Triatoma dimidiata is the vector of Trypanosoma cruzi in the Yucatan Peninsula (YP). Earlier studies have shown that domestic and peri‐domestic populations of the vector originated from the sylvan stock and that effectiveness of insecticide‐spraying was affected by re‐infestations of houses from the sylvan T. dimidiata population. In addition, in the YP most previously published reports have focused on domestic and peri‐domestic populations and very little is known about the nocturnal behavior of the sylvan populations. The main aim of our study was to determine the nightly activity patterns of adult T. dimidiata in a selected location in the YP. Secondly, we sought to document the reproductive status and infection rate of active females. During eight sampling nights spaced from late March to late July, 2007, we collected 544 adult T. dimidiata. We found that square‐cloth illuminated white traps were effective to attract the sylvan individuals and that T. dimidiata adults exhibited a unimodal activity pattern throughout the night. The accumulated mean of captured bugs also showed a non‐linear distribution for females and males. Furthermore, we found that male and female catches were significantly correlated with the means of temperature and humidity recorded during the sampling period. Out of 46 dissected females, we observed that 43.5% of females had fully‐formed eggs in their abdomens, and only two females (4.4%) had sperm within the spermatheca. The infection rate of T. dimidiata harboring T. cruzi was found to be 3.7%. The implications of the light attraction to bugs and potential dispersal capabilities are discussed in the paper in the context of infestation/re‐infestation of rural houses by sylvan T. dimidiata flying adults.     

Independent and joint action of cis- and trans-permethrin in Triatoma infestans (Hemiptera: Reduviidae)

The toxicity of pure cis- and trans-permethrin or mixtures of the two isomers topically applied to first, third, and fifth instar nymphs of Triatoma infestans (Klug) at 26°C was determined. The cis-isomer was more active than the trans-isomer in the three stages evaluated. When the two isomers were simultaneously applied to first instar nymphs, an additive effect was observed. Similar treatments of third and fifth instar nymphs resulted in an antagonistic effect. In third instar nymphs, the cis-isomer was more active than trans-isomer at all the three temperatures assayed (16°, 26°, and 36°C). The toxicity of the cis-isomer was lower at 36°C than at either 16° or 26°C. Temperature had no significant effect on the toxicity of the trans-isomer within the temperature range assayed. The toxicity of either isomer to third instar nymphs was not affected by pretreatment of nymphs with PBO (an inhibitor of mixed-function oxidases activity) or TPP (an inhibitor of esterase activity), suggesting that these detoxification pathways are not relevant in the metabolism of cis- or trans-isomers. Arch. Insect Biochem. Physiol. 37:225–230, 1998. © 1998 Wiley-Liss, Inc.     

Factors associated with the occurrence of Triatoma sordida (Hemiptera: Reduviidae) in rural localities of Central-West Brazil

This study estimates the factors of artificial environments (houses and peridomestic areas) associated with Triatoma sordida occurrence. Manual searches for triatomines were performed in 136 domiciliary units (DUs) in two rural localities of Central-West Brazil. For each DU, 32 structural, 23 biotic and 28 management variables were obtained. Multiple logistic regression analysis was performed in order to identify statistically significant variables associated with occurrence of T. sordida in the study areas. A total of 1,057 specimens (99% in peridomiciles, mainly chicken coops) of T. sordida were collected from 63 DUs (infestation: 47%; density: ~8 specimens/DU; crowding: ~17 specimens/infested DU; colonisation: 81%). Only six (0.6%) out of 945 specimens examined were infected with Trypanosoma cruzi. The final adjusted logistic regression model indicated that the probability of T. sordida occurrence was higher in DU with wooden chicken coops, presence of > 30 animals in wooden corrals, presence of wood piles and presence of food storeroom. The results show the persistence of T. sordida in peridomestic habitats in rural localities of Central-West Brazil. However, the observed low intradomestic colonisation and minimal triatomine infection rates indicate that T. sordida has low potential to sustain high rates of T. cruzi transmission to residents of these localities.     

Isolation and characterization of polymorphic microsatellite loci in the Chagas’ disease vector Triatoma infestans (Hemiptera: Reduviidae)

Microsatellites were identified and characterized from Triatoma infestans, the principal vector of Chagas’ disease in the Southern Cone of South American countries. Ninety‐three microsatellite loci were isolated from partial genomic libraries, of which 30 were amplified and 10 were selected for genotyping. The degree of intrapopulation variation in these loci was determined using 34 specimens from the locality of Chancaní (Córdoba, Argentina). The number of alleles per locus ranged from five to 19 and the expected heterozygosity ranged from 0.608 to 0.941. The variability of these microsatellite markers provides a valuable molecular tool for population genetic studies in T. infestans.     

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.     

Fine structure of the salivary glands of Triatoma infestans (Hemiptera: Reduviidae)

The fine structure of the salivary glands of adult Triatoma infestans (Hemiptera: Reduviidae) bugs has been analyzed. Stereomicroscopy and scanning electron microscopy showed that each insect presents a pair of salivary glands, each pair containing three distinct units (main, supplementary, and accessory) with different sizes and colors. Transmission electron microscopy demonstrated that all gland units consist of a monolayer of epithelial cells surrounding a large central lumen. The gland units are enveloped by a thick basal lamina containing bundles of muscle cells. Microvilli are present at the apical plasma membrane domain of the gland cells, thus enlarging the available membrane area for saliva secretion towards the large gland lumen, although occasionally budding vesicles could be observed among the microvilli. Cytochemical analysis showed that the salivary gland cells of T. infestans present abundant endoplasmic reticulum profiles and several lipid droplets.     

Life history of two abundant populations of Dipetalogaster maximus (Uhler, 1894) (Hemiptera: Reduviidae: Triatominae) in northwestern Mexico

We focused on the analysis of biological parameters of two different abundant populations of Dipetalogaster maximus (Uhler) (Hemiptera: Reduviidae: Triatominae) from northwestern Mexico. The biological parameters were related to hatching of eggs, life cycle, and meal needed for molting by each instar of two populations from environmentally similar areas (El Fandango and San Dionisio) with a similar number of available hosts as blood meal sources. The D. maximus populations from the two locations were evaluated and compared. No significant differences (P>0.05) were recorded for the average hatching time of the two cohorts. The median egg‐to‐adult development time and the number of blood meals at each nymphal group were significantly shorter (P<0.05) for the El Fandango cohort. The number of obtained females at the end of the cycles, number of eggs laid, and rate of egg hatching were significantly higher (P<0.05) for the El Fandango cohort. These results contribute to the estimation of abundances of the studied populations of D. maximus in areas where many tourists go for camping and have an increased risk of being bitten and infected by this species.     

Genetic structure of Triatoma venosa (Hemiptera: Reduviidae): molecular and morphometric evidence

Triatoma venosa presents a restricted geographical distribution in America and is considered as a secondary vector of Chagas disease in Colombia and Ecuador. A total of 120 adult insects were collected in domestic and peridomestic habitats in an endemic area of the department of Boyacá, Colombia, in order to determine their genetic structure through morphometric and molecular techniques. The head and wings of each specimen were used for the analyses of size, shape, and sexual dimorphism. A significant sexual dimorphism was found, although no differences in size among the studied groups were detected. Differences were found in the analyzed structures except for male heads. DNA was extracted from the legs in order to carry out the internal transcriber space-2 (ITS-2) amplification and the randon amplified polymorphic DNA (RAPD) analyses. Length polymorphisms were not detected in the ITS-2. Fst and Nm values were estimated (0.047 and 3.4, respectively). The high genetic flow found among the insects captured in the domicile and peridomiciliary environment does not permit a genetic differentiation, thus establishing the peridomicile as an important place for epidemiological surveillance.     

Comparison of insecticidal paint and deltamethrin against Triatoma infestans (Hemiptera: Reduviidae) feeding and mortality in simulated natural conditions

The vector of Chagas disease, Triatoma infestans, is largely controlled by the household application of pyrethroid insecticides. Because effective, large‐scale insecticide application is costly and necessitates numerous trained personnel, alternative control techniques are badly needed. We compared the residual effect of organophosphate‐based insecticidal paint (Inesfly 5A IGR? (I5A)) to standard deltamethrin, and a negative control, against T. infestans in a simulated natural environment. We evaluated mortality, knockdown, and ability to take a blood meal among 5^th instar nymphs. I5A paint caused significantly greater mortality at time points up to nine months compared to deltamethrin (Fisher's Exact Test, p < 0.01 in all instances). A year following application, mortality among nymphs in the I5A was similar to those in the deltamethrin (χ2 = 0.76, df=1, p < 0.76). At months 0 and 1 after application, fewer nymphs exposed to deltamethrin took a blood meal compared to insects exposed to paint (Fisher's Exact Tests, p < 0.01 and p < 0.01, respectively). Insecticidal paint may provide an easily‐applied means of protection against vectors of Chagas disease.