Misidentification of two Brazilian triatomes, Triatoma arthurneivai and Triatoma wygodzinskyi, revealed by geometric morphometrics

Triatoma arthurneivai Lent & Martins and Triatoma wygodzinskyi Lent (Hemiptera: Reduviidae) are two Brazilian species found in the sylvatic environment. Several authors may have misidentified T. arthurneivai and consequently published erroneous information. This work reports the use of geometric morphometric analysis on wings in order to differentiate T. arthurneivai and T. wygodzinskyi, and thus to detect possible misidentifications. Triatomines collected from the field in the states of Minas Gerais and São Paulo, and from laboratory colonies, were used. Analyses show a clear differentiation between specimens of T. arthurneivai and T. wygodzinskyi. This indicates that T. arthurneivai populations from São Paulo state were misidentified and should be considered as T. wygodzinskyi. This study also suggests that T. arthurneivai is an endemic species from Serra do Cipó, Minas Gerais state.     

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.     

First genotyping of Trypanosoma cruzi from naturally infected Triatoma juazeirensis,Triatoma melanica and Triatoma sherlocki from Bahia State,Brazil

Many previous studies have shown a great phylogenetic and biological variability of Trypanosoma cruzi using different molecular and biochemical methods. Populations of T. cruzi were initially clustered into two main lineages called TcI and TcII by the size of the mini‐exon PCR product. In the present study, 33 isolates derived from three triatomine taxa, which belong to the Triatoma brasiliensis species complex (Triatoma juazeirensis, Triatoma melanica and Triatoma sherlocki); collected in three distinct areas of Bahia state were characterized by PCR. The isolates were identified by the size of the mini‐exon gene, 18S rRNA and 24Sα rRNA amplicons. T. cruzi isolates obtained in sylvatic and intradomiciliar ecotopes, derived from T. juazeirensis and T. melanica, were identified as TcI while the parasites originated from T. sherlocki were characterized as TcI and TcII genotypes, respectively. Those species are present in sylvatic ecotopes but are able to infest intradomiciliar areas. Therefore, it would be important to maintain studies in those localities of Bahia and further investigate the possibilities of Chagas disease transmission. Human disease may occur by any T. cruzi genotype and not only by TcII as it is the case in Amazonia.     

Volatile compounds emitted by Triatoma dimidiata,a vector of Chagas disease: chemical analysis and behavioural evaluation

In this study, we evaluated the responses of Triatoma dimidiata Latreille (Hemiptera: Reduviidae) to volatiles emitted by conspecific females, males, mating pairs and metasternal gland (MG) extracts with a Y‐tube olfactometer. The volatile compounds released by mating pairs and MGs of T. dimidiata were identified using solid‐phase microextraction and coupled gas chromatography‐mass spectrometry (GC‐MS). Females were not attracted to volatiles emitted by males or MG extracts; however, they preferred clean air to their own volatiles or those from mating pairs. Males were attracted to volatiles emitted by males, females, mating pairs, pairs in which the male had the MG orifices occluded or MG extracts of both sexes. However, males were not attracted to volatiles emitted by pairs in which the female had the MG orifices occluded. The chemical analyses showed that 14 and 15 compounds were detected in the headspace of mating pairs and MG, respectively. Most of the compounds identified from MG except for isobutyric acid were also detected in the headspace of mating pairs. Both females and males were attracted to octanal and 6‐methyl‐5‐hepten‐2‐one, and males were attracted to 3,5‐dimethyl‐2‐hexanol. Males but not females were attracted to a seven‐compound blend, formulated from compounds identified in attractive MG extracts.     

Recovery of Triatoma infestans populations after insecticide application: an experimental field study

The capacity of populations of Triatoma infestans (Klug) (Hemiptera: Reduviidae) to survive and recover was assessed after application of insecticide (gamma-HCH at a rate of 0.5 g a.i./m2) at different seasons. T. infestans populations were maintained in experimental chicken houses under natural climatic conditions in a region of Argentina endemic for Chagas disease transmitted by these bugs. Based on previous studies of T. infestans populations in these habitats, each experimental group was set up with a total of 626 T. infestans, comprising 390 eggs, 204 nymphs of particular stages, fourteen male and eighteen female adults. The chicken houses were dismantled and rebuilt at monthly intervals to study the vector population changes over a period of 33 months. When the insecticide was applied during winter, spring or summer, populations of T. infestans recovered to untreated or precontrol levels during the next reproductive season (i.e. during the hot season, October-March). In contrast, populations treated during autumn (March) remained at very low densities for 2 years and then increased rapidly to surpass the untreated populations. All populations of the bugs fell to very low numbers (sometimes less than twenty individuals) after gamma-HCH applications, but none was driven to extinction. In all cases, the density of surviving populations was independent of their density before treatment. The fact that all treated populations recovered within 1-3 years, to at least the density of untreated populations, shows the high reproductive potential of T. infestans to recover from very low population densities. Moreover, the additive effect of climatic-induced mortality and insecticide-induced mortality is only apparent when insecticides are applied just before the onset of the cold winter months during which reproductive rates are at their lowest.     

Morphological and morphometric analysis of scutella of six species and two subspecies of Triatoma (Hemiptera: Reduviidae) in Mexico

Morphological characters can be used to distinguish the vast majority of triatomine species, but the existence of high levels of phenotypic plasticity and recently diverged species can lead to erroneous determinations. To approach this problem, we analyzed the male and female morphologies of the scutella of Triatoma barberi, T. dimidiata, T. lecticularia, T. mexicana, T. recurva, T. rubida, and two sub‐species, T. protracta protracta and T. protracta nahuatlae. Scutellum samples were observed by scanning electron microscopy and subjected to morphological analysis and morphometric investigation using a canonical discriminant analysis. The results revealed differences primarily in central depression shape, posterior process, and vestiture. We observed clear dimension‐based differences in scutellum morphometry in all the taxa under study, providing sound evidence for species and subspecies differentiation. On the other hand, there is no difference between sexes in T. lecticularia, T. protracta protracta, and T. protracta nahuatlae. Our methodology can be implemented to differentiate species of the genus Triatoma.     

Population dynamics and control of Triatoma infestans

Analysis of field populations of Triatoma infestans (Hemiptera: Reduviidae), after a 3-year study, shows that population growth rate is affected by both density-dependent and density-independent mortality. Although an equilibrium exists, apparently as a consequence of a density dependent-mechanism, population density fluctuates throughout the year because of the effect of monthly mean minimum temperature as a density-independent source of mortality. Simulation studies based on Moran curves shows that high population densities have an approximately constant extinction probability (around 0.20), independently of the season the population starts growing. However, at very low population densities, the extinction probability depends strongly on the season when the population begins to grow. Very low density populations beginning in winter or autumn have the highest extinction probability. The outcome of the simulation studies coincides with results observed in field populations affected by insecticide application at different seasons.     

Ecology of Triatoma brasiliensis in northeastern Brazil: seasonal distribution,feeding resources,and Trypanosoma cruzi infection in a sylvatic population

We assessed some ecological parameters of Triatoma brasiliensis in rock piles in the state of Ceará during the rainy and dry seasons. The greatest density was in April (median = 12.5 triatomines/site). The greatest abundance was in December, when the insects were more dispersed and the density per site was lower (6 triatomines/site). The nutritional status of females and 5^th instar nymphs was increased in July. The rate of T. cruzi infection reached its highest peak in July (10.9%). ELISA revealed that the principal food sources were birds (33.1%), followed by armadillos (18.8%). Food sources were more frequently identified during the rainy season. T. brasiliensis specimens collected in the drought tended to: i) present lower rates of T. cruzi infection and gut content reactivity to tested antisera, ii) have a poorer nutritional status, iii) exhibit lower fecundity, iv) be more dispersed among the studied collection sites, and v) be more abundant and easily collected in the surface of the rocks, possibly reflecting an increased searching for blood meals. Such findings underscore epidemiological concerns and allow inferences about the season when triatomines can more frequently invade the peridomestic environment in search of food and recolonize artificial structures.     

Population structure of the Chagas disease vector,Triatoma infestans,at the urban–rural interface

The increasing rate of biological invasions resulting from human transport or human‐mediated changes to the environment has had devastating ecological and public health consequences. The kissing bug, Triatoma infestans, has dispersed through the Peruvian city of Arequipa. The biological invasion of this insect has resulted in a public health crisis, putting thousands of residents of this city at risk of infection by Trypanosoma cruzi and subsequent development of Chagas disease. Here, we show that populations of Tria. infestans in geographically distinct districts within and around this urban centre share a common recent evolutionary history although current gene flow is restricted even between proximal sites. The population structure among the Tria. infestans in different districts is not correlated with the geographical distance between districts. These data suggest that migration among the districts is mediated by factors beyond the short‐range migratory capabilities of Tria. infestans and that human movement has played a significant role in the structuring of the Tria. infestans population in the region. Rapid urbanization across southern South America will continue to create suitable environments for Tria. infestans, and knowledge of its urban dispersal patterns may play a fundamental role in mitigating human disease risk.     

Comparison of two sampling methods for domestic populations of Triatoma infestans in north-west Argentina

Abstract. In mud-and-thatch houses of Santiago del Estero Province, northwest Argentina, where no insecticides had been sprayed officially to control domestic infestations of the Chagas disease vector Triatoma infestans (Klug) (Hemiptera: Reduviidae), two knockdown (KD) insecticidal collection procedures were compared with the standard flush-out (FO) method for sampling T. infestans. Bugs were collected by FO using 0.2% tetramethrin in bedrooms of (1) 41 houses of Amama village employing 1 man-hour of capture effort per house, and (2) 19 houses of Trinidad and Mercedes villages employing 4 man-hours/house. From the same houses, 2-5 days after the manual FO collection, bugs were collected by KD indoor-spraying of deltamethrin 25mga.i./m² in Amama, or burning of one γ-HCH (=γ-BHC) fumigant tablet 3.1g of γ-isomer) per bedroom in Trinidad and Mercedes. The majority of infestations were detected by both methods, the proportion of positive houses being 81% at Amama and 95% at the other villages. Although the FO method was more sensitive, at least because it was applied first, the KD method detected infestations in 25% of houses where bugs were not found by FO. Bug densities estimated by FO or by subsequent KD in each house were significantly correlated: r = 0.795 for deltamethrin; r= 0.882 for y-HCH. Compared with FO collections of T.infestans large stages, i.e. adults plus fourth and fifth instar nymphs, the KD catch averaged 0.88x with deltamethrin and 0.57× with γ-HCH, regardless of the apparent population density of bugs per house. However, the KD method has practical advantages of speed and standardization.     

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.”     

Determinants of the domiciliary density of Triatoma infestans, vector of Chagas disease

In two heavily infested rural villages of Santiago del Estero, Argentina, where no indoor-spraying with residual insecticides had ever been carried out by official control services, we studied the influence of roof and wall structure, domestic use of insecticide, family size and the number of domestic dogs, on the domiciliary density of Triatoma infestans (Klug). Bug density was significantly associated with (1) the interaction between insecticide use and type of roof, (2) the structure of indoor walls, (3) the number of dogs sharing sleeping areas of people (room-mate dogs), and (4) the number of people plus room-mate dogs, but not with just the number of people resident in the house. The interaction between insecticide use and a roof made of 'simbol', a locally available grass (Pennisetum sp.), also reflected a younger age structure of domestic bug populations. In infested houses, the density of bugs infected with Trypanosoma cruzi Chagas was significantly correlated with overall bug density. Our data suggest that the application of environmental management measures by the affected people, such as plastering of walls and modification of roofs, coupled with keeping dogs away from bedrooms and application of insecticides, should limit the domestic population density of T. infestans and thus reduce the transmission of T. cruzi to people.     

Population differentiation of the Chagas disease vector Triatoma maculata (Erichson, 1848) from Colombia and Venezuela

The emerging vector of Chagas disease, Triatoma maculata (Hemiptera, Reduviidae), is one of the most widely distributed Triatoma species in northern South America. Despite its increasing relevance as a vector, no consistent picture of the magnitude of genetic and phenetic diversity has yet been developed. Here, several populations of T. maculata from eleven Colombia and Venezuela localities were analyzed based on the morphometry of wings and the mitochondrial NADH dehydrogenase subunit 4 (ND4) gene sequences. Our results showed clear morphometric and genetic differences among Colombian and Venezuelan populations, indicating high intraspecific diversity. Inter‐population divergence is suggested related to East Cordillera in Colombia. Analyses of other populations from Colombia, Venezuela, and Brazil from distinct eco‐geographic regions are still needed to understand its systematics and phylogeography as well as its actual role as a vector of Chagas disease.     

Peridomestic distribution of Triatoma garciabesi and Triatoma guasayana in north-west Argentina

The Reduviid bugs Triatoma garciabesi Carcavallo et al. (Hemiptera: Reduviidae: Triatominae), previously known as T. sordida Ståhl in the semi‐arid chaco region, and T. guasayana Wygodzinsky & Abalos, vectors of Trypanosoma cruzi Chagas (Kinetoplastida: Trypanosomatidae), were found to occupy wide but different ranges of ecotopes in the peridomestic environment. At Amamá and nearby rural villages in north‐western Argentina, a combined total of 1233 specimens were collected from 325/2314 (14%) sites surveyed at 6‐monthly intervals from November 1994 to November 1996. Triatoma garciabesi and T. guasayana displayed a significantly different distribution among peridomestic ecotopes. Triatoma garciabesi predominated over T. guasayana in the prevalence of infested sites, the number of colonies and the number of bugs collected. For T. garciabesi, the predominant ecotopes most likely to yield T. garciabesi repeatedly were the rugged bark of Prosopis alba or P. nigra (Fabaceae) trees, where chickens roosted, and chicken coops. For T. guasayana the main ecotopes were goat or sheep corrals, piled materials and orchard fences. Triatoma garciabesi and T. guasayana were collected concurrently from the same site on 9/2314 (0.4%) occasions, and on different dates at the same site on 12 (0.5%) occasions. The observed low frequency of mixed populations (< 1%) was not significantly different from that expected from a hypothesis of independence. Triatoma garciabesi clearly outnumbered T. guasayana in four of the nine mixed populations, none of which persisted as such. Neither T. garciabesi nor T. guasayana colonized human habitations, even in the absence of T. infestans (formerly the predominant domestic vector of T. cruzi in this area), a situation that apparently has not changed in the last 50 years in northern Argentina.     

Excretion/defecation patterns in Triatoma infestans populations that are,respectively, susceptible and resistant to deltamethrin

Pyrethroid resistance has been detected in Triatoma infestans (Klug) (Hemiptera: Reduviidae) specimens from different areas of Argentina and Bolivia. Genes conferring resistance can have a pleiotropic effect with epidemiological and evolutionary consequences. This research studied excretion/defecation patterns in deltamethrin‐resistant T. infestans in order to elucidate its biological performance, adaptive consequences and role in the transmission of Chagas' disease. One deltamethrin‐susceptible strain and two deltamethrin‐resistant strains were used. Fifth‐instar nymphs were fed ad libitum and their defecations recorded during and after the first or second feeding in the stadium. Resistant insects began to defecate later, defecated less, showed a lower proportion of defecating individuals and lower defecation indices compared with susceptible insects during the first hour after feeding. The number of bloodmeals in the stadium did not affect the main variables determining the pattern of defecation. The present study suggests that alterations in the excretion/defecation pattern in resistant insects entail an adaptive cost and, considering only this pattern, determine a lower capacity for transmission of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) compared with susceptible insects.     

Micropropagation in banana using high levels of cytokinins does not involve any genetic changes as revealed by RAPD and ISSR markers

Use of high levels of growth regulators during micropropagation results in undesirable clonal variability in important commercial crops such as banana. The present study investigated the effects of high levels of cytokinins on micropropagation in banana (genotype AAB), and the genetic stability of plantlets was assessed using RAPD and ISSR markers. Cytokinins, such as BA and kinetin were added to the routine shoot multiplication medium at concentrations up to 10 mg l⁻¹. After 12 weeks of culture involving three subcultures, the maximum number of shoot buds were produced in cultures receiving either 5 mg l⁻¹ BA (80 shoot buds) or 4 mg l⁻¹ kinetin (62 shoot buds). Certain morphological abnormalities observed during proliferation of shoot buds in vitro were not observed during acclimatization ex vitro. To check the genetic stability, RAPD and ISSR profiles of micropropagated plantlets obtained from different cytokinin-treatments were compared with control microplants maintained on MS medium as well as the field-grown mother plant. A total of 50 RAPD and 12 ISSR primers resulted in 625 distinct and reproducible bands. Thus a total of 17,400 bands were generated showing homogeneous RAPD and ISSR patterns. Band intensity histogram of each gel confirmed their monomorphic nature with no genetic variation in all the plantlets analysed. Based on these results a protocol for high rate shoot multiplication was worked out leading to uniform shoot production.     

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.     

Clonal and spatial genetic structure in natural populations of Luohanguo (Siraitia grosvenorii), an economic species endemic to South China,as revealed by RAPD markers

Clonal growth is generally expected to have significant effects on the spatial genetic structure within populations. In this study, random amplified polymorphic DNA (RAPD) markers were used to reveal clonal and spatial genetic structure of four natural populations of Luohanguo (Siraitia grosvenorii), an economic vine species endemic to South China. A total of 351 ramets were assigned to 76 distinct multi-locus genotypes (i.e. genets), with the G/N varying from 0.121 to 0.350. No widespread genet was found across different populations. The clonal diversity (D) and evenness (E) ranged from 0.333 to 0.828 and from 0 to 0.741, respectively. While most genets consisted of fewer than five ramets, we observed some dominant genets that had much more (up to 69) ramets and spread over large areas. Spatial autocorrelation analyses revealed a spatial genetic structure (i.e. significant positive autocorrelation within 20 m and negative autocorrelation beyond 40 m) in one population, but not in other three populations with smaller population size. This study highlights the importance of clonal growth in shaping the spatial genetic structure in Luohanguo, which may have complex effects on the dynamics and evolution of its declining populations.     

Effects of chickens on the prevalence of infestation and population density of Triatoma infestans in rural houses of north-west Argentina

Abstract Effects of the presence of chickens on population density of the bug Triatoma infestans , principal vector of Chagas disease, were investigated by standardized sampling (indoors and peridomestically) from sixty-eight houses of three rural communities in north-west Argentina, during March 1992.
The domiciliary density of T. infestans increased linearly with the percentage of bugs that fed on chickens, as identified by agar double-diffusion tests. Bug density was significantly higher in houses where hens usually nested indoors than in those where they did not, as determined by concurrent direct observations and interviews of householders. Multiple linear regression analysis of domiciliary bug density on (a) the total number of people, dogs and cats per house; (b) the percentage of domiciliary bugs that fed on chickens, or (c) the indoor-brooding habit of hens, showed the two variables (b, c) related to chickens as significant predictors in each regression model. Inclusion both variables representing chickens increased the fit significantly. Addition of other potentially confounding factors (domestic insecticide use, type of roof and walls of house) did not affect the significant variables retained in the best-fitting regression model.
Peridomestic infestation was positively associated with the household number of fowls. Chickens were the main bloodmeal source of peridomestic T.infestans populations. Human-fed bugs were detected in peridomestic sites of sixteen houses, indicating active dispersal of adults and large nymphal instars from bedroom areas.
Exclusion of hens from domiciliary areas and promotion of chicken sheds, of an appropriate design that would not harbour bug populations, should limit the triatomine population growth rate and reduce the risk of infestation.     

The morphological variation of the eggs and genital plates of two morphotypes of Triatoma protracta Uhler, 1894

The control of triatomine insects is necessary because these insects are the principal vectors of Trypanosoma cruzi, the agent of Chagas disease. Nevertheless, some of these vectors, such as Triatoma protracta, have not been studied adequately and their importance and taxonomic status has not yet been defined in detail and must be reevaluated in view of the continuing taxonomic uncertainties associated with the species. To help clarify the taxonomic status of T. protracta, the eggs and genital plates of two morphotypes were analyzed. Qualitative and quantitative morphological differences were observed in two morphotypes, designated T. p. protracta and T. p. nahuatlae according to Ryckman (1962). The morphotype T. p. protracta exhibited large and wide eggs with pores forming large padded polygonal structures, whereas the eggs of the morphotype T. p. nahuatlae were small and smooth. The size of the 9(th) genital urosternite was longer and wider in females in contrast to males in both morphotypes. However, these size differences were relatively greater in T. p. protracta. The high morphological variation found between the morphotypes of T. protracta suggests that they should be separated. Accordingly, it is probable that this group should be re-classified.