Variety is the spice: The role of morphological variation of Triatoma infestans (Hemiptera, Reduviidae) at a macro-scale.

Chagas disease is caused by the protozoan parasite Trypanosoma cruzi (Chagas, 1909). One of the primary vectors of T. cruzi in South America is Triatoma infestans (Klug, 1834). This triatomine species is distributed across a huge latitudinal gradient, inhabiting domiciliary , peridomiciliary , and wild environments. Its wide geographic distribution provides an excellent opportunity to study the relationships between environmental gradients and intraspecific morphological variation. In this study, we investigated variations in wing size and shape in T. infestans across six ecoregions. We aimed to address the following questions: How do wing size and shape vary on a regional scale, does morphological variation follow specific patterns along an environmental or latitudinal gradient, and what environmental factors might contribute to wing variation? Geometric morphometric methods were applied to the wings of 162 females belonging to 21 T. infestans populations, 13 from Argentina (n = 105), 5 from Bolivia (n = 42), and 3 from Paraguay (n = 15). A comparison of wing centroid size across the 21 populations showed significant differences. Canonical Variate Analysis (CVA) revealed significant differences in wing shape between the populations from Argentina, Bolivia, and Paraguay, although there was a considerable overlap, especially among the Argentinian populations. Well-structured populations were observed for the Bolivian and Paraguayan groups. Two analyses were performed to assess the association between wing size and shape, geographic and climatic variables: multiple linear regression analysis (MRA) for size and Partial Least Squares (PLS) regression for shape. The MRA showed a significant general model fit. Six temperature-related variables, one precipitation-related variable, and the latitude showed significant associations with wing size. The PLS analysis revealed a significant correlation between wing shape with latitude, longitude, temperature-related, and rainfall-related variables. Wing size and shape in T. infestans populations varied across geographic distribution. Our findings demonstrate that geographic and climatic variables significantly influence T. infestans wing morphology.

An increased number of individuals of a potential host facilitates non-photic synchronisation in the haematophagous insect Triatoma infestans.

BACKGROUND: Triatoma infestans (Kissing bug) is the main vector of the parasite causative of Chagas disease in Latin-America. This species shows clear activity rhythms easily synchronised to day-night cycles (photic cycle). The haematophagous nature of these insects lead us to think that they may temporally adapt to the particular activity rhythms of potential hosts (non-photic cycle). Our previous data showed that kissing bugs were weakly affected by the activity-inactivity rhythm of a single host. OBJETIVE: To determine if by increasing the number of individuals of a potential host, T. infestans could increase the likelihood of synchronisation. METHODS: Individual activity rhythms of experimental insects, maintained in constant darkness in light-tight cabinets, localised in a room with 24 rodents, were continuously monitored. Another insect group that served as control was maintained in the same conditions but in a room without rodents. FINDINGS: Most of the experimental insects synchronised, expressing a 24 h period coincident with the activity-inactivity rhythms of the rodents, while the controls free ran with a period significantly longer than 24 h. CONCLUSION: Analogous to what happens with high vs low light intensity in photic synchronisers, a high number of rodents, in contrast to the previous one-rodent experiment, increased the potency of this non-photic zeitgeber.

Phenotypic variability in traits related to flight dispersal in the wing dimorphic species Triatoma guasayana.

BACKGROUND: Triatoma guasayana is considered an emerging vector of Chagas disease in the Southern Cone of South America. The presence of a triatomine population with brachypterous individuals, in which both wings are reduced, has recently been reported for this species. The aim of the present study was to determine if flight-related traits varied across populations, if these traits could explain differences in flight capacity across populations and if flight-related traits are associated with geographic and/or climatic variation. METHODS: The study involved 66 male T. guasayana specimens from 10 triatomine populations. Digital images of wing, head and pronotum were used to estimate linear and geometric morphometric variables. Variations in size and shape were analysed using one-way analysis of variance and canonical variate analysis (CVA), respectively. Mantel tests were applied to analyse the relationship between morphometric and geographic distances, and the association between size measurements was analysed using Pearson's correlation. We explored covariation between size and shape variables using partial least square analyses (PLS). The association of geographic and climatic variables with size measurements was tested using linear regression analyses. We performed PLS analyses for shape measurements. RESULTS: Wing size differed significantly across triatomine populations. The CVA showed that wing shape of the brachypterous population is well discriminated from that of the other populations. The Mantel test showed a positive and significant association between wing shape and geographic distances. The heads of the brachypterous population were significantly larger than those of the other populations. Similar to wing shape, the head shape of the brachypterous population was well discriminated from those of the other populations. Pronotum width did not show significant differences across populations. Geographic and climatic factors were associated with size and shape of both the wing and head, but not with pronotum width. CONCLUSIONS: Most of the traits related to flight dispersal varied across populations. Wing shape and head shape were found to be better markers for differentiated morphological variation across populations. Head measurements also varied in accordance with this condition. Geographic and climatic variables were associated with most of the flight-related traits.

An increased number of individuals of a potential host facilitates non-photic synchronisation in the haematophagous insect Triatoma infestans

BACKGROUND Triatoma infestans (Kissing bug) is the main vector of the parasite causative of Chagas disease in Latin-America. This species shows clear activity rhythms easily synchronised to day-night cycles (photic cycle). The haematophagous nature of these insects lead us to think that they may temporally adapt to the particular activity rhythms of potential hosts (non-photic cycle). Our previous data showed that kissing bugs were weakly affected by the activity-inactivity rhythm of a single host. OBJETIVE To determine if by increasing the number of individuals of a potential host, T. infestans could increase the likelihood of synchronisation. METHODS Individual activity rhythms of experimental insects, maintained in constant darkness in light-tight cabinets, localised in a room with 24 rodents, were continuously monitored. Another insect group that served as control was maintained in the same conditions but in a room without rodents. FINDINGS Most of the experimental insects synchronised, expressing a 24 h period coincident with the activity-inactivity rhythms of the rodents, while the controls free ran with a period significantly longer than 24 h. CONCLUSION Analogous to what happens with high vs low light intensity in photic synchronisers, a high number of rodents, in contrast to the previous one-rodent experiment, increased the potency of this non-photic zeitgeber.

History of insecticide resistance of Triatominae vectors

In the last 15 years, different types of Triatominae resistance to different insecticides have been reported; thus, resistance may be more widespread than known, requiring better characterization and delimitation, which was the aim of this review. This review was structured on a literature search of all articles from 1970 to 2015 in the PubMed database that contained the keywords Insecticide resistance and Triatominae. Out of 295 articles screened by title, 33 texts were selected for detailed analysis. Insecticide resistance of Triatomines is a complex phenomenon that has been primarily reported in Argentina and Bolivia, and is caused by different factors (associated or isolated). Insecticide resistance of Triatominae is a characteristic inherited in an autosomal and semi-dominant manner, and is polygenic, being present in both domestic and sylvatic populations. The toxicological profi le observed in eggs cannot be transposed to different stages of evolution. Different toxicological profi les exist at macro- and microgeographical levels. The insecticide phenotype has both reproductive and developmental costs. Different physiological mechanisms are involved in resistance. Studies of Triatomine resistance to insecticides highlight three defi ciencies in interpreting the obtained results: I) the vast diversity of methodologies, despite the existence of a single guiding protocol; II) the lack of information on the actual impact of resistance ratios in the fi eld; and III) the concept of the susceptibility reference lineage. Research on the biological and behavioral characteristics of each Triatominae species that has evolved resistance is required in relation to the environmental conditions of each region.

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.

Rural houses infestation by Triatoma infestans in northwestern Argentina: Vector control in a high spatial heterogeneous infestation area.

Triatoma infestans (Hemiptera: Reduviidae) is a vector of the Trypanosoma cruzi parasite, causative agent of Chagas disease. During the last decade, vector control activities have been systematically carried out in northwestern Argentina, an endemic region for this disease. The general aim of this study to evaluate was spatio-temporal variation of infestation by T. infestans in rural communities of Los Llanos in La Rioja province. We estimated house infestation using two sampling methods: passive and active. Passive collection was conducted with community participation collecting triatomines. Six passive collections were carried out in 397 houses during the warm season between 2014 and 2017. Active collection of T. infestans was thoroughly performed by trained staff for 60 minutes and was carried out once in March 2016. The estimate of intradomestic infestation did not show significant differences between both collection methods (p = 0.39). However, passive collection method had lower sensitivity than active collection method for the estimation of peridomestic infestation and intradomestic colonization (PDI: p< 0.01; ID colonization: p< 0.01). The results obtained with passive collection methods showed that the infestation in the study area was spatially heterogeneous and temporally variable. Intradomiciliary infestation decreased over time (14.4% to 7.9%, p<0.05) although the effect of the chemical treatment application was not associated with the infestation level of T. infestans (p = 0.15) and the Departments had a different response each year (p<0.01). A high infestation cluster was located in the south of our study area during 2016-2017. The vector presence in the houses confirms the importance of to improve entomological surveillance programs. The search for triatomines carried out by the inhabitants might be a useful method to complement the activities of vector control programs in isolated and rural areas.

Circadian system responses to nocturnal and diurnal hosts in the kissing bug, Triatoma infestans.

Insects express diverse behavioral rhythms synchronized to environmental cycles. While circadian entrainment to light-dark cycles is ubiquitous in living organisms, synchronization to non-photic cycles may be critical for hematophagous bugs that depend on rhythmic hosts. The purpose was to determine whether Triatoma infestans are capable of synchronizing to the circadian rhythms of potential hosts with temporally distinct activity patterns; and, if so, if this synchronization occurs through masking or entrainment. Precise synchronization with the food source may be critical for the insects' survival due to the specific predatory or defensive nature of each host. Kissing bugs were housed in a compartment in constant dark, air-flow-connected to another compartment with a nocturnal or a diurnal host; both hosts were synchronized to a light-dark cycle. The activity rhythms of kissing bugs were modulated by the daily activity rhythms of the vertebrates. Effects were a decrease in the endogenous circadian period, independent of the host being nocturnal or diurnal; in some cases relative coordination occurred and in others synchronization was clearly achieved. Moreover, splitting and bimodality arose, phenomena that were also affected by the host presence. The results indicate that T. infestans were able to detect the non-photic cycle of their potential hosts, an ability that surely facilitates feeding and hinders predation risk. Understanding triatomines behavior is of fundamental importance to the design of population control methods.

Morphometric variability among the species of the Sordida subcomplex (Hemiptera: Reduviidae: Triatominae): evidence for differentiation across the distribution range of Triatoma sordida.

BACKGROUND: The Sordida subcomplex (Triatominae) comprises four species, Triatoma garciabesi, T. guasayana, T. patagonica and T. sordida, which differ in epidemiological importance and adaptations to human environments. Some morphological similarities among species make taxonomic identification, population differentiation and species delimitation controversial. Triatoma garciabesi and T. sordida are the most similar species, having been considered alternatively two and a single species until T. garciabesi was re-validated, mostly based on the morphology of male genitalia. More recently, T. sordida from Argentina has been proposed as a new cryptic species distinguishable from T. sordida from Brazil, Bolivia and Paraguay by cytogenetics. We studied linear and geometric morphometry of the head, wings and pronotum in populations of these species aiming to find phenotypic markers for their discrimination, especially between T. sordida and T. garciabesi, and if any set of variables that validates T. sordida from Argentina as a new species. RESULTS: Head width and pronotum length were the linear variables that best differentiated species. Geometric morphometry revealed significant Mahalanobis distances in wing shape between all pairwise comparisons. Triatoma patagonica exhibited the best discrimination and T. garciabesi overlapped the distribution of the other species in the morphometric space of the first two DFA axes. Head shape showed differentiation between all pairs of species except for T. garciabesi and T. sordida. Pronotum shape did not differentiate T. garciabesi from T. guasayana. The comparison between T. garciabesi and T. sordida from Argentina and T. sordida from Brazil and Bolivia revealed low differentiation based on head and pronotum linear measurements. Pronotum and wing shape were different between T. garciabesi and T. sordida from Brazil and Bolivia and T. sordida from Argentina. Head shape did not differentiate T. garciabesi from T. sordida from Argentina. CONCLUSIONS: Wing shape best delimited the four species phenotypically. The proposed cryptic species, T. sordida from Argentina, differed from T. sordida from Brazil and Bolivia in all measured shape traits, suggesting that the putative new species may not be cryptic. Additional studies integrating cytogenetic, phenotypic and molecular markers, as well as cross-breeding experiments are needed to confirm if these three entities represent true biological species.

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.

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

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.

Dispersión de triatoma infestans y otros triatominos en el Chaco árido de la Argentina / Spread of triatoma infestans and other triatomines in the arid Chaco in Argentina

Nuestro estudio comprueba la existencia de dispersión activa de adultos y ninfas de T. infestans caminando y confirma la dispersión por vuelo de la especie.

Dispersal of Triatoma infestans and other Triatominae species in the arid Chaco of Argentina: flying, walking or passive carriage? The importance of walking females.

The aim of this paper was to analyse the active dispersal of Triatoma infestans and the role of chickens as passive carriers of this insect in peridomestic areas of La Rioja, Argentina. To measure active dispersal, monthly catches were made on six consecutive nights for five months (in the warm season) using light traps (for flying insects) and sticky dispersal barriers (for walking insects). The nutritional and reproductive states of adults were evaluated. Over the course of the sampling period, a total of eight flying adults, six walking nymphs and 10 walking adults of the species T. infestans were captured, as well as specimens of Triatoma guasayana, Triatoma eratyrusiformis and Triatoma platensis. Our data demonstrate for the first time that females of T. infestans can disperse by walking. This may be an adaptive strategy because it allows them to move with eggs and/or with good blood reserves, which are not possible when flying. All flying and walking individuals of both genders were of an appropriate physiological state that would allow for colonisation of the target habitat. However, manual inspection of 122 chickens suggests that it is unlikely that these animals passively transport T. infestans. Finally, the dispersal activity of T. infestans was compared with other triatomines using a dispersion index.

Dispersal of Triatoma infestans and other Triatominae species in the arid Chaco of Argentina: Flying, walking or passive carriage? The importance of walking females

The aim of this paper was to analyse the active dispersal of Triatoma infestans and the role of chickens as passive carriers of this insect in peridomestic areas of La Rioja, Argentina. To measure active dispersal, monthly catches were made on six consecutive nights for five months (in the warm season) using light traps (for flying insects) and sticky dispersal barriers (for walking insects). The nutritional and reproductive states of adults were evaluated. Over the course of the sampling period, a total of eight flying adults, six walking nymphs and 10 walking adults of the species T. infestans were captured, as well as specimens of Triatoma guasayana, Triatoma eratyrusiformis and Triatoma platensis. Our data demonstrate for the first time that females of T. infestans can disperse by walking. This may be an adaptive strategy because it allows them to move with eggs and/or with good blood reserves, which are not possible when flying. All flying and walking individuals of both genders were of an appropriate physiological state that would allow for colonisation of the target habitat. However, manual inspection of 122 chickens suggests that it is unlikely that these animals passively transport T. infestans. Finally, the dispersal activity of T. infestans was compared with other triatomines using a dispersion index.

Phenotypic variability and population structure of peridomestic Triatoma infestans in rural areas of the arid Chaco (Western Argentina): spatial influence of macro- and microhabitats.

Triatoma infestans, one of the main vectors of Trypanosoma cruzi, is strongly associated with rural human ecotopes. Infested peridomestic places could act as a source of house infestation as a result of the movement of vectors among habitats. The quantitative study of phenotypic traits and nutritional variables could be informative about the population structure and the relative mobility of vectors. This is the first approach toward analyzing the phenetic characteristics of T. infestans and the relationship with their potential role to invade and colonize other habitats in the arid Chaco region. This article compares the phenotypic structure of T. infestans populations living in chicken coops and goat corrals, analyzing the geometric morphometry of wings and heads, antennal phenotypes, and the nutritional status of 257 adults and nymphs. The study area remained untreated with insecticides during the 15 years before the present study. The insects collected in peridomestic structures located in two zones 80 km apart exhibited significant differences in their morphotypes, suggesting an environmental effect and/or the absence of individual exchange between zones. The population structure was clear in T. infestans from goat corrals and chicken coops in one zone and less pronounced in the other. These results suggest the existence of a different rate of individual exchange among ecotopes within each zone. Morphometric variables of heads and wings were significantly correlated in insects collected in goat corrals but not in chicken coops, suggesting a habitat effect and supporting the hypothesis of different canalization forces that affect the two organs. The nutritional status of adults indicated a low dispersal probability with zonal differences. This article provides the first combined morphometric analysis of the head of adults and fifth instar nymphs in triatomines and the first combined analysis of head and wing in T. infestans.

Phenotypic variability associated to genomic changes in the main vector of Chagas disease in the southern cone of South America.

This study shows for the first time, the relationship between chromosomic and meristic quantitative characters of Triatoma infestans, the main vector of Chagas disease in the southern cone of South America. The antennal phenotype of 225 adults from five countries and three cytotypes, were analyzed using univariate and multivariate techniques. The study found a clear association between cytotypes and antennal phenotypes. The complexity of the antennal sensilla pattern was higher in the non-Andean cytotype, showing more chemoreceptors usually linked to sexual behaviour, host finding and flight. The phenetic differences were more notorious on females than on males. Changes in the sexual dimorphism were detected, being the differences most notorious in the non-Andean cytotype. Similitude of the geographic populations using the antennal phenotype coincides with environmental gradient of the area, showing a close association between the physical features of the living zones and quantitative traits of the vectors. The epidemiological implications of the phenetic differences observed in the study are discussed.