Comparative Study of the Antennal Phenotype in Species of the Belminus (Hemiptera: Triatominae) Genus Using Optical and Scanning Electron Microscopy.

The genus Belminus Stål, 1859 is distinguished by markedly entomophagous species. This genus is a poorly studied group of the Triatominae (Jeannel, 1919), which includes hematophagous species that are vectors of Trypanosoma cruzi (Chagas, 1909), the etiological agent of Chagas disease. This study reports for the first time the description of the antennal sensilla of Belminus corredori (Galvão & Angulo, 2003), Belminus herreri (Lent & Wygodzinsky, 1979), and Belminus ferroae (Sandoval, Pabón, Jurberg & Galvão, 2007) nymphs and adults throughout scanning electron microscopy and optical microscopy. Nine morphological types of sensilla were identified in the three species, including bristles I and II, trichobothria, tapered hairs, thin-walled trichoid, thick-walled trichoid, basiconic, campaniform, and coeloconica sensilla. The analysis of the most abundant types of sensilla and their distribution in nymphs and adults shows similarities with the antennal phenotypes described in other Triatominae. Quantitative differences showed the important phenotypic plasticity of this morphological trait in the genus and allowed for the separation of the species in both sexes. These differences were associated with sexual dimorphism and wing condition (macropterous and brachypterous). The role of the antennal phenotype in sexual behavior, its usefulness as a taxonomic tool, and its relationship with the dispersal capacity of these species are discussed.

Activity levels of female Triatoma infestans change depending on physiological condition.

BACKGROUND: Triatoma infestans (Hemiptera: Reduviidae) is the main vector of Trypanosoma cruzi, the etiological agent of Chagas disease, in South America. Active dispersal of this vector is the principal cause of recolonization of human dwellings previously treated with insecticides. Due to the persistence of vector populations and their movement between habitats, dispersive behavior studies are important for understanding the epidemiology of Chagas disease. The aim of this study was to analyze the relationship among T. infestans females' activity levels according to their physiological conditions. RESULTS: Two groups of insects were used, unfed and fed females. Each was composed of three subgroups in relation to the reproductive state: fifth-stage nymphs, virgin and fertilized females. There was a significant interaction between reproductive and nutritional states among T. infestans female' activity levels. During the experiments, unfed and fed nymphs remained inactive. Virgin females showed a dual behavior in their movement; fasted insects were more active. Fertilized females, both fed and unfed, were always active. CONCLUSION: The reproductive and nutritional conditions of T. infestans females affect their activity levels. When females with different reproductive states remained together, fertilized females showed permanent activity levels, suggesting that this subgroup of females represents the highest epidemiological risk as colonizers of human dwellings.

Active dispersal of Triatoma infestans and other triatomines in the Argentinean arid Chaco before and after vector control interventions.

Peridomestic structures are considered the main sites where Triatoma infestans (Hemiptera: Reduviidae) remain and disperse, representing the first risk factor for intradomestic invasion, even after vector control activities. This study analyzed T. infestans dispersal during vector control interventions in six rural houses of the arid Chaco (La Rioja, Argentina). Flying and walking dispersers were captured during five months of two consecutive warm seasons after insecticide spraying of intra- and peridomiciles. These data were compared with previous published data in the same scenario but without insecticide spraying in peridomiciles. Recorded climatic conditions were favorable for active dispersion during the study. Total number of T. infestans dispersers moving among domestic habitats decreased after insecticide spraying. Sylvatic triatomines T. guasayana, T. eratyrusiformis, T. garciabesi, and T. platensis, not targeted by insecticide spraying, were captured simultaneously within peridomestic areas and showed higher invasion pressure than T. infestans. Adult T. infestans peridomestic populations showed high nutritional status, indicating low dispersion probability. Some peridomiciles remained infested at the end of the study. However, no intradomiciles were recolonized. These results suggest that there is a low probability of intradomestic recolonization by active dispersion from peridomiciles during 15 months post-spraying.

Antennal phenotype of Mexican haplogroups of the Triatoma dimidiata complex, vectors of Chagas disease.

Triatoma dimidiata (Latreille) is a species complex that spans North, Central, and South America and which is a key vector of all known discrete typing units (DTU) of Trypanosoma cruzi, the etiologic agent of Chagas disease. Morphological and genetic studies indicate that T. dimidiata is a species complex with three principal haplogroups (hg) in Mexico. Different markers and traits are still inconclusive regarding if other morphological differentiation may indicate probable behavioral and vectorial divergences within this complex. In this paper we compared the antennae of three Mexican haplogroups (previously verified by molecular markers ND4 and ITS-2) and discussed possible relationships with their capacity to disperse and colonized new habitats. The abundance of each type of sensillum (bristles, basiconics, thick- and thin-walled trichoids) on the antennae of the three haplogroups, were measured under light microscopy and compared using Kruskal-Wallis non-parametric and multivariate non-parametric analyses. Discriminant analyses indicate significant differences among the antennal phenotype of haplogroups either for adults and some nymphal stages, indicating consistency of the character to analyze intraspecific variability within the complex. The present study shows that the adult antennal pedicel of the T. dimidiata complex have abundant chemosensory sensilla, according with good capacity for dispersal and invasion of different habitats also related to their high capacity to adapt to conserved as well as modified habitats. However, the numerical differences among the haplogroups are suggesting variations in that capacity. The results here presented support the evidence of T. dimidiata as a species complex but show females and males in a different way. Given the close link between the bug's sensory system and its habitat and host-seeking behavior, AP characterization could be useful to complement genetic, neurological and ethological studies of the closely related Dimidiata Complex haplogroups for a better knowledge of their vectorial capacity and a more robust species differentiation.

Can body traits, other than wings, reflect the flight ability of Triatominae bugs?

INTRODUCTION: Insects of the subfamily Triatominae are vectors of Trypanosoma cruzi , the Chagas disease parasite, and their flying behavior has epidemiological importance. The flying capacity is strikingly different across and within Triatominae species, as well as between sexes or individuals. Many Triatoma infestans individuals have wings but no flying muscles. In other Triatominae species, no clear relationships were found between wing length and flying behavior. If wing presence or size is not reflective of the flying behavior, which other parts of the body could be considered as reliable markers of this important function? METHODS: The genus Mepraia has exceptional characteristics with invariably wingless females and wingless or winged males. We calculated the porous surface exposed to odorant molecules to estimate the olfactory capacity of Mepraia spinolai . The head shape and thorax size were estimated using the geometric morphometric approach and traditional morphometric techniques, respectively. RESULTS: Alary polymorphism in M. spinolai was significantly associated with consistent modification of the thorax size, head shape, and notable change in the estimated olfactory capacity. The macropterous individuals had a larger olfactory surface and thorax size and significantly different head shape compared to those of the micropterous individuals. CONCLUSIONS: We concluded that these structural changes could be associated with the flying potential of Triatominae. Thus, morphological attributes not found on wings could help determine the likely flying potential of the bugs.

Can body traits, other than wings, reflect the flight ability of Triatominae bugs?

Abstract: INTRODUCTION : Insects of the subfamily Triatominae are vectors of Trypanosoma cruzi , the Chagas disease parasite, and their flying behavior has epidemiological importance. The flying capacity is strikingly different across and within Triatominae species, as well as between sexes or individuals. Many Triatoma infestans individuals have wings but no flying muscles. In other Triatominae species, no clear relationships were found between wing length and flying behavior. If wing presence or size is not reflective of the flying behavior, which other parts of the body could be considered as reliable markers of this important function? METHODS : The genus Mepraia has exceptional characteristics with invariably wingless females and wingless or winged males. We calculated the porous surface exposed to odorant molecules to estimate the olfactory capacity of Mepraia spinolai . The head shape and thorax size were estimated using the geometric morphometric approach and traditional morphometric techniques, respectively. RESULTS : Alary polymorphism in M. spinolai was significantly associated with consistent modification of the thorax size, head shape, and notable change in the estimated olfactory capacity. The macropterous individuals had a larger olfactory surface and thorax size and significantly different head shape compared to those of the micropterous individuals. CONCLUSIONS: We concluded that these structural changes could be associated with the flying potential of Triatominae. Thus, morphological attributes not found on wings could help determine the likely flying potential of the bugs.

Thermal preferences and limits of Triatoma brasiliensis in its natural environment--field observations while host searching.

The goal of this work was to explore the thermal relationship between foraging Triatoma brasiliensis and its natural habitat during the hottest season in the state of Ceará, Brazil. The thermal profiles were determined using infrared analysis. Although the daily temperature of rock surfaces varied in a wide range, T. brasiliensis selected to walk through areas with temperatures between 31.7-40.5ºC. The temperature of T. brasiliensis body surface ranged from 32.8-34.4ºC, being higher in legs than the abdomen. A strong relationship was found between the temperature of the insect and the temperature of rock crevices where they were hidden (r: 0.96, p < 0.05). The species was active at full sunlight being a clear example of how the light-dark rhythm may be altered, even under predation risk. Our results strongly suggest a thermal borderline for T. brasiliensis foraging activity near 40ºC. The simultaneous determination of insect body and rock temperatures here presented are the only obtained in natural habitats for this or other triatomines.

Thermal preferences and limits of Triatoma brasiliensis in its natural environment - Field observations while host searching

The goal of this work was to explore the thermal relationship between foraging Triatoma brasiliensis and its natural habitat during the hottest season in the state of Ceará, Brazil. The thermal profiles were determined using infrared analysis. Although the daily temperature of rock surfaces varied in a wide range, T. brasiliensisselected to walk through areas with temperatures between 31.7-40.5ºC. The temperature of T. brasiliensisbody surface ranged from 32.8-34.4ºC, being higher in legs than the abdomen. A strong relationship was found between the temperature of the insect and the temperature of rock crevices where they were hidden (r: 0.96, p < 0.05). The species was active at full sunlight being a clear example of how the light-dark rhythm may be altered, even under predation risk. Our results strongly suggest a thermal borderline for T. brasiliensisforaging activity near 40ºC. The simultaneous determination of insect body and rock temperatures here presented are the only obtained in natural habitats for this or other triatomines.

Control of rural house infestation by Triatoma infestans in the Bolivian Chaco using a microencapsulated insecticide formulation.

BACKGROUND: Triatoma infestans, the main vector of Trypanosoma cruzi (causative agent of Chagas disease) has been successfully eliminated over much of its original geographic distribution over the southern cone countries of South America. However, populations of the species are still infesting houses of rural communities of the Gran Chaco region of Argentina and Bolivia. This study reports for the first time a large-scale effect of a vector control intervention using a microencapsulated formulation of organophosphates and insect growth regulator on house infestation by T. infestans, in the southwestern region of Santa Cruz de la Sierra Department, within the Bolivian chaco. METHODS: The vector control intervention included the treatment and entomological evaluation of 1626 individually coded and georeferenced houses with the microencapsulated formulation. House infestation by T. infestans was evaluated by active searches with fixed capture effort carried out before and after two, 16 and 32 months of the treatment application. RESULTS: House infestation prevalence was 30.5% before the intervention, spatially aggregated in two clusters of 38 and 25 localities that showed 41% and 38% house infestation by T. infestans. Infestation prevalence was reduced to 2.4% two months after the intervention and remained very low (1.7%) until the end of the study after 32 months of the control intervention, without any other additional vector control intervention. CONCLUSIONS: The obtained results show an important long lasting effect on house protection against triatomine infestation in a region of known pyrethroid resistant populations of T. infestans, as the result of the slow release of the active ingredients, protected by the formulation microcapsule.

Effect of feeding frequency on the reproductive efficiency of two species of Triatoma with different epidemiological importance.

INTRODUCTION: In Triatominae, reproductive efficiency is an important factor influencing population dynamics, and a useful parameter in measuring a species' epidemiological significance as a vector of Trypanosoma cruzi (Chagas, 1909). The reproductive efficiency of triatomines is affected by food availability; hence, we measured and compared the effects of feeding frequency on the reproductive parameters of Triatoma patagonica (Del Ponte, 1929) and Triatoma infestans (Klug, 1934), and the effects of starvation on T. patagonica. METHODS: Couples from both species were fed weekly, or every 3 weeks; in addition, females in couples of T. patagonica were not fed. Each couple was observed weekly and reproductive efficiency was assessed on the following parameters: fecundity (eggs/female), fertility (eggs hatched/eggs laid), initiation and end of oviposition, initiation of mating, number of matings/week, and number of reproductive weeks. Relative meal size index (RMS), blood consumption index (CI), and E values (eggs/mg blood) were also calculated. RESULTS: Changes in feeding frequency affected the reproductive parameters of T. patagonica only, with a decrease in fecundity and number of reproductive weeks for those fed every 3 weeks, or not fed. The reproductive period, RMS index, and CI were lower for T. patagonica than T. infestans. However, despite the lower fecundity of T. patagonica, this species required less blood to produce eggs, with an E values of 2 compared to 2.94 for T. infestans. CONCLUSIONS: Our results suggest that the differences in fecundity observed between species reflect the availability of food in their natural ecotopes.

Effect of feeding frequency on the reproductive efficiency of two species of Triatoma with different epidemiological importance

Introduction In Triatominae, reproductive efficiency is an important factor influencing population dynamics, and a useful parameter in measuring a species' epidemiological significance as a vector of Trypanosoma cruzi (Chagas, 1909). The reproductive efficiency of triatomines is affected by food availability; hence, we measured and compared the effects of feeding frequency on the reproductive parameters of Triatoma patagonica (Del Ponte, 1929) and Triatoma infestans (Klug, 1934), and the effects of starvation on T. patagonica. Methods Couples from both species were fed weekly, or every 3 weeks; in addition, females in couples of T. patagonica were not fed. Each couple was observed weekly and reproductive efficiency was assessed on the following parameters: fecundity (eggs/female), fertility (eggs hatched/eggs laid), initiation and end of oviposition, initiation of mating, number of matings/week, and number of reproductive weeks. Relative meal size index (RMS), blood consumption index (CI), and E values (eggs/mg blood) were also calculated. Results Changes in feeding frequency affected the reproductive parameters of T. patagonica only, with a decrease in fecundity and number of reproductive weeks for those fed every 3 weeks, or not fed. The reproductive period, RMS index, and CI were lower for T. patagonica than T. infestans. However, despite the lower fecundity of T. patagonica, this species required less blood to produce eggs, with an E values of 2 compared to 2.94 for T. infestans. Conclusions Our results suggest that the differences in fecundity observed between species reflect the availability of food in their natural ecotopes. .

Circadian entrainment by light and host in the Chagas disease vector, Triatoma infestans.

Triatoma infestans (Reduviidae: Triatominae, "kissing bug") is the main insect vector of Trypanosoma cruzi, the causative agent of Chagas disease, a chronic trypanosomiasis infecting 10 million people world-wide. This hematophagous bug feeds on diurnal and nocturnal species during each host's quiescent time. As the hosts are also its major predators, kissing bugs are subjected to dual selective pressures from a single source. Therefore, synchronization of feeding with the host's behavior is critical to the insects' survival. We show that nonphotic signals linked to the host eclipse the role of light and dark as the primary circadian zeitgeber for these bugs, although light still strongly inhibits locomotor behavior directly. In nature, this combination provides the insect with great flexibility in organizing physiology and behavior: anticipating a quiescent host or avoiding its potential predation while remaining directly responsive to immediate environmental conditions. Manipulation of nonphotic entrainment could be a useful chronobiotic tool in the control of Chagas disease.

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.

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.

Potential sources of Triatoma infestans reinfesting peridomiciles identified by morphological characterization in Los Llanos, La Rioja, Argentina.

The presence of Triatoma infestans in habitats treated with insecticides constitutes a frequent problem in endemic areas. Basing our study on the hypothesis that descendants of a residual population should be more similar to the pre-treatment population than to any other, we compared the indications of two quantitative morphological approaches. This study seeks to find the origin of 247 T. infestans from three populations found in two chicken coops and a goat corral after treatment with insecticides. The results obtained by quantitative morphology suggest that the T. infestans found between three-34 months after the application of insecticides formed mixed populations with insects derived from residual foci and neighbouring habitats. Our analyses also showed the presence of a phenotype which does not resemble neither the pre-treatment phenotype nor the one from neighbouring populations, suggesting the presence of a particular post-treatment phenotype. The heads size showed some variations in males from different populations and remained unchanged in females, which reinforces the hypothesis of an intraspecific competition for food with priority for females. This article presents, for the first time, the combined analysis of geometric morphometry of heads and antennal phenotypes to identify the composition of reinfesting populations.

Potential sources of Triatoma infestans reinfesting peridomiciles identified by morphological characterization in Los Llanos, La Rioja, Argentina

The presence of Triatoma infestans in habitats treated with insecticides constitutes a frequent problem in endemic areas. Basing our study on the hypothesis that descendants of a residual population should be more similar to the pre-treatment population than to any other, we compared the indications of two quantitative morphological approaches. This study seeks to find the origin of 247 T. infestans from three populations found in two chicken coops and a goat corral after treatment with insecticides. The results obtained by quantitative morphology suggest that the T. infestans found between three-34 months after the application of insecticides formed mixed populations with insects derived from residual foci and neighbouring habitats. Our analyses also showed the presence of a phenotype which does not resemble neither the pre-treatment phenotype nor the one from neighbouring populations, suggesting the presence of a particular post-treatment phenotype. The heads size showed some variations in males from different populations and remained unchanged in females, which reinforces the hypothesis of an intraspecific competition for food with priority for females. This article presents, for the first time, the combined analysis of geometric morphometry of heads and antennal phenotypes to identify the composition of reinfesting populations.

Changes related to gender, geographic population and habitat in the antennal phenotype of Triatoma patagonica Del Ponte, 1929 (Hemiptera: Reduviidae).

Triatomines undergo morphological changes as an adaptive response to different habitats (wild, peridomestic, domestic, laboratory). The characterization of the antennal phenotype provides information on intraspecific variation caused by geographical origin and/or habitat. Triatoma patagonica Del Ponte, 1929 is known to occur in peridomiciles of rural areas in Argentina, where it also invades non-colonized dwellings. Here we describe and compare the antennal phenotype of T. patagonica in populations of different geographic origin, and explore possible modifications induced by laboratory rearing with the aim of investigating the range of phenotypic variation of the species for the first time. Sixty antennas of adult males and females of T. patagonica belonging to two peridomiciliary populations of different geographical origin were analyzed. Four types of sensilla were observed in three antennal segments, showing sexual dimorphism in the species. The multivariate analysis separated the populations of similar habitat (peridomicile) but different geographical origin, without showing differences between the peridomiciliary and laboratory populations of the same geographical origin. These results suggest phenotypic plasticity in T. patagonica, which would allow the species to adapt to a wide range of habitats without having a close association with a given host and its environment. The range of antennal phenotypic variation of T. patagonica would also be an indicator of its current stage of adaptation to the human environment.

The residual efficacy of a cypermethrin pour-on formulation applied on goats on the mortality and blood intake of Triatoma infestans

Triatoma infestans is the main vector of Trypanosoma cruzi, the aetiological agent of Chagas disease in the Gran Chaco region of South America. As a frequent blood meal source for triatomine bugs, domestic goats play a key role in the eco-epidemiology of Chagas disease. The aim of this study was to evaluate the mortality and blood intake of T. infestans fed on goats that had been treated with different doses of pour-on insecticide. Third-instar nymphs were fed on goats that had been treated with 0 cc, 5 cc, 10 cc or 15 cc of a pour-on formulation of cypermethrin. The exposure of T. infestans to animals treated at different post-application intervals revealed a residual activity of the insecticide. The mortality rate in the treated groups was higher than in the control groups until 30 days post-insecticide application (p = 0.03), except in the group treated with 5 cc, in which no mortality was detected after seven days of insecticide application. Rainfall affected the triatomicide effect, reducing the time of residual activity. The cypermethrin pour-on treatment decreased the blood intake of T. infestans. Thirty days after the cypermethrin application, nymph mortality was 16% (± 13) with both doses (10 cc and 15 cc). The 15 cc dose did not result in higher insect mortality or increased persistence compared to the 10 cc dose.

Effects of fipronil on dogs over Triatoma infestans, the main vector of Trypanosoma cruzi, causative agent of Chagas disease.

Chagas disease is the most important endemic disease in Latin America, mainly transmitted by Triatoma infestans in the Southern Cone countries of South America. Dogs are one of the main domestic reservoirs of Trypanosoma cruzi, the etiological agent of Chagas disease. The presence of dogs in rural households of endemic areas significantly increases the likelihood of the vectorial transmission of the parasite. We studied the mortality and blood intake of T. infestans exposed to dogs treated with different doses and formulations of fipronil. Two doses, two formulations, and different distances to the application point of fipronil were compared. Third instar nymphs of T. infestans were fed at different time intervals after the insecticide application up to 45 days post-application. No significant difference was found between the blood intake of nymphs fed on control and treated dogs with different doses and formulations (p > 0.05). The spray formulation showed lower effect and persistence than the spot-on formulation. The mortality rate caused by the spot-on formulation in the 26.8-mg active ingredient (a.i.)/kg dose was higher (48%) than with the 13.4-mg a.i./kg dose (25%), 24 h after the insecticide application. The effect was highly heterogeneous among replicates of the same treatment. The mortality rate of nymphs fed over the point of the insecticide application was higher than the mortality of nymphs fed over places 12 cm apart from the fipronil application point, suggesting that the distribution of fipronil over the dog body is lower than the needed one to obtain a persistent triatomicide effect.

Exploring the association between Trypanosoma cruzi infection in rural communities and environmental changes in the southern Gran Chaco.

The association between land use and land cover changes between 1979-2004 in a 2.26-million-hectare area south of the Gran Chaco region and Trypanosoma cruzi infection in rural communities was analysed. The extent of cultural land, open and closed forests and shrubland up to 3,000 m around rural communities in the north, northwest and west of the province of Córdoba was estimated using Landsat satellite imagery. The T. cruzi prevalence was estimated with a cross-sectional serological survey conducted in the rural communities. The land cover showed the same patterns in the 1979, 1999 and 2004 satellite imagery in both the northwest and west regions, with shrinking regions of cultured land and expanding closed forests away from the community. The closed forests and agricultural land coverage in the north region showed the same trend as in the northwest and west regions in 1979 but not in 1999 or 2004. In the latter two years, the coverage remote from the communities was either constant or changed in opposite ways from that of the northwest and west regions. The changes in closed forests and cultured vegetation alone did not have a significant, direct relationship with the occurrence of rural communities with at least one person infected by T. cruzi. This study suggests that the overall decrease in the prevalence of T. cruzi is a consequence of a combined effect of vector control activities and changes in land use and land cover.