Population Structure of the Chagas Disease Vector Triatoma infestans in an Urban Environment

Chagas disease is a vector-borne disease endemic in Latin America. Triatoma infestans, a common vector of this disease, has recently expanded its range into rapidly developing cities of Latin America. We aim to identify the environmental features that affect the colonization and dispersal of T. infestans in an urban environment. We amplified 13 commonly used microsatellites from 180 T. infestans samples collected from a sampled transect in the city of Arequipa, Peru, in 2007 and 2011. We assessed the clustering of subpopulations and the effect of distance, sampling year, and city block location on genetic distance among pairs of insects. Despite evidence of genetic similarity, the majority of city blocks are characterized by one dominant insect genotype, suggesting the existence of barriers to dispersal. Our analyses show that streets represent an important barrier to the colonization and dispersion of T. infestans in Arequipa. The genetic data describe a T. infestans infestation history characterized by persistent local dispersal and occasional long-distance migration events that partially parallels the history of urban development.     

Intensified Surveillance and Insecticide-based Control of the Chagas Disease Vector Triatoma infestans in the Argentinean Chaco

Background

The elimination of Triatoma infestans, the main Chagas disease vector in the Gran Chaco region, remains elusive. We implemented an intensified control strategy based on full-coverage pyrethroid spraying, followed by frequent vector surveillance and immediate selective insecticide treatment of detected foci in a well-defined rural area in northeastern Argentina with moderate pyrethroid resistance. We assessed long-term impacts, and identified factors and procedures affecting spray effectiveness.

Methods and Findings

After initial control interventions, timed-manual searches were performed by skilled personnel in 4,053 sites of 353–411 houses inspected every 4–7 months over a 35-month period. Residual insecticide spraying was less effective than expected throughout the three-year period, mainly because of the occurrence of moderate pyrethroid resistance and the limited effectiveness of selective treatment of infested sites only. After initial interventions, peridomestic infestation prevalence always exceeded domestic infestation, and timed-manual searches consistently outperformed householders'' bug detection, except in domiciles. Most of the infestations occurred in houses infested at baseline, and were restricted to four main ecotopes. Houses with an early persistent infestation were spatially aggregated up to a distance of 2.5 km. An Akaike-based multi-model inference approach showed that new site-level infestations increased substantially with the local availability of appropriate refugia for triatomine bugs, and with proximity to the nearest site found infested at one or two preceding surveys.

Conclusions and Significance

Current vector control procedures have limited effectiveness in the Gran Chaco. Selective insecticide sprays must include all sites within the infested house compound. The suppression of T. infestans in rural areas with moderate pyrethroid resistance requires increased efforts and appropriate management actions. In addition to careful, systematic insecticide applications, housing improvement and development policies that improve material conditions of rural villagers and reduce habitat suitability for bugs will contribute substantially to sustainable vector and disease control in the Gran Chaco.     

Strong Host-Feeding Preferences of the Vector Triatoma infestans Modified by Vector Density: Implications for the Epidemiology of Chagas Disease

Background

Understanding the factors that affect the host-feeding preferences of triatomine bugs is crucial for estimating transmission risks and predicting the effects of control tactics targeting domestic animals. We tested whether Triatoma infestans bugs prefer to feed on dogs vs. chickens and on dogs vs. cats and whether vector density modified host choices and other vital rates under natural conditions.

Methodology

Two host choice experiments were conducted in small caged huts with two rooms between which bugs could move freely. Matched pairs of dog–chicken (six) and dog–cat (three) were assigned randomly to two levels of vector abundance and exposed to starved bugs during three nights. Bloodmeals from 1,160 bugs were tested by a direct enzyme-linked immunosorbent assay.

Principal Findings

Conditional logistic regression showed that dogs were highly preferred over chickens or cats and that vector density modified host-feeding choices. The relative risk of a bug being blood-engorged increased significantly when it fed only on dog rather than chicken or cat. Bugs achieved higher post-exposure weight at higher vector densities and successive occasions, more so if they fed on a dog rather than on a cat.

Conclusions

Our findings strongly refute the hypothesis that T. infestans prefers to blood-feed on chickens rather than dogs. An increase in dog or cat availability or accessibility will increase the rate of bug feeding on them and exert strong non-linear effects on R ₀. When combined with between-dog heterogeneities in exposure, infection, and infectiousness, the strong bug preference for dogs can be exploited to target dogs in general, and even the specific individuals that account for most of the risk, with topical lotions or insecticide-impregnated collars to turn them into baited lethal traps or use them as transmission or infestation sentinels based on their immune response to Trypanosoma cruzi or bug salivary antigens.     

Host-Seeking Behavior and Dispersal of Triatoma infestans,a Vector of Chagas Disease,under Semi-field Conditions

Chagas disease affects millions of people in Latin America. The control of this vector-borne disease focuses on halting transmission by reducing or eliminating insect vector populations. Most transmission of Trypanosoma cruzi, the causative agent of Chagas disease, involves insects living within or very close to households and feeding mostly on domestic animals. As animal hosts can be intermittently present it is important to understand how host availability can modify transmission risk to humans and to characterize the host-seeking dispersal of triatomine vectors on a very fine scale. We used a semi-field system with motion-detection cameras to characterize the dispersal of Triatoma infestans, and compare the behavior of vector populations in the constant presence of hosts (guinea pigs), and after the removal of the hosts. The emigration rate – net insect population decline in original refuge – following host removal was on average 19.7% of insects per 10 days compared to 10.2% in constant host populations (p = 0.029). However, dispersal of T. infestans occurred in both directions, towards and away from the initial location of the hosts. The majority of insects that moved towards the original location of guinea pigs remained there for 4 weeks. Oviposition and mortality were observed and analyzed in the context of insect dispersal, but only mortality was higher in the group where animal hosts were removed (p-value <0.01). We discuss different survival strategies associated with the observed behavior and its implications for vector control. Removing domestic animals in infested areas increases vector dispersal from the first day of host removal. The implications of these patterns of vector dispersal in a field setting are not yet known but could result in movement towards human rooms.     

Cost-Effectiveness of Chagas Disease Vector Control Strategies in Northwestern Argentina

Background

Control and prevention of Chagas disease rely mostly on residual spraying of insecticides. In Argentina, vector control shifted from a vertical to a fully horizontal strategy based on community participation between 1992 and 2004. The effects of such strategy on Triatoma infestans, the main domestic vector, and on disease transmission have not been assessed.

Methods and Findings

Based on retrospective (1993–2004) records from the Argentinean Ministry of Health for the Moreno Department, Northwestern Argentina, we performed a cost-effectiveness (CE) analysis and compared the observed CE of the fully horizontal vector control strategy with the expected CE for a vertical or a mixed (i.e., vertical attack phase followed by horizontal surveillance) strategy. Total direct costs (in 2004 US$) of the horizontal and mixed strategies were, respectively, 3.3 and 1.7 times lower than the costs of the vertical strategy, due to reductions in personnel costs. The estimated CE ratios for the vertical, mixed and horizontal strategies were US$132, US$82 and US$45 per averted human case, respectively. When per diems were excluded from the costs (i.e., simulating the decentralization of control activities), the CE of vertical, mixed and horizontal strategies was reduced to US$60, US$42 and US$32 per averted case, respectively.

Conclusions and Significance

The mixed strategy would have averted between 1.6 and 4.0 times more human cases than the fully horizontal strategy, and would have been the most cost-effective option to interrupt parasite transmission in the Department. In rural and dispersed areas where waning vertical vector programs cannot accomplish full insecticide coverage, alternative strategies need to be developed. If properly implemented, community participation represents not only the most appealing but also the most cost-effective alternative to accomplish such objectives.     

The biting rate of Triatoma infestans in Argentina

1. The daily proportion of fed individuals in a population of the reduviid bug Triatoma infestans (Klug), maintained under natural climatic conditions in experimental chicken houses in central Argentina, was estimated from the proportion of bugs retaining colourless urine in the rectum. 2. From the estimates of feeding frequency throughout a 1 year period, it was shown that temperature has a dominant effect on biting rate, but density-dependent effects became apparent during the warmest months. 3. These and other data on the determinants of blood consumption by T. infestans, were incorporated into a detailed hypothesis of density regulation in this species.     

Triatoma infestans in Greater Buenos Aires, Argentina

The Health Administration Agencies of many municipalities in Greater Buenos Aires (GBA) receive frequent reports on triatomines in houses. The aim of this work was to identify and describe the dispersal foci of Triatoma infestans in an urban neighborhood of GBA, and contribute to the knowledge of the epidemiological situation in the region. In June 1998, potentially infested places were entomologically evaluated. T. infestans was only detected in a hen building for egg production, which housed approximately 6,000 birds. A total of 2,930 insects were collected. Density was about 9 triatomines/m(2). The proportions of fifth instar nymphs and adults were significantly higher than those of the other stages (p<0.001). The number of triatomines collected largely exceeded the highest domestic infestation found in one house from rural endemic areas of Argentina. Though triatomines were negative for Trypanosoma cruzi, they could acquire the parasite by coming in contact with infected people living in GBA. Besides, the numerous and widely distributed places housing hens and chickens, would favor the settlement of the vector. Together, both facts may constitute a risk of parasitic vectorial transmission. It is recommended to intensify systematic activities of vector search and case detection in GBA.     

Demographic effects of deltamethrin resistance in the Chagas disease vector Triatoma infestans

Triatoma infestans (Heteroptera: Reduviidae) Klug is the main vector of Chagas disease in Latin America. Resistance to deltamethrin was reported in Argentina and recently associated with reproductive and longevity trade‐offs. The objectives of the present study were to describe the demographic consequences of deltamethrin resistance in T. infestans and to establish possible target stages for chemical control in susceptible and resistant colonies. A stage‐classified matrix model was constructed based on the average stage length for susceptible, resistant and reciprocal matings' progeny. The differences between colonies were analysed by prospective and retrospective analysis. The life table parameters indicated reduced fecundity, fertility and population growth in resistant insects. The retrospective analysis suggested the latter was associated with lower reproductive output and increased fifth‐instar nymph stage length. The prospective analysis suggested that the adult stage should be the main target for insecticide control. Although, fifth‐instar nymphs should also be targeted when resistance has been detected. The presented results show demographic effects of deltamethrin resistance in T. infestans. While the older stages could be the main targets for chemical control, this approach is impeded by their higher tolerance to insecticides. It is concluded that the different mode of action insecticides would be more effective than a dose increase for the control of deltamethrin‐resistant T. infestans.     

Molecular phylogeography of the Chagas' disease vector Triatoma infestans in Argentina

Triatoma infestans is the main vector of Chagas' disease in South America between latitudes 10°S and 46°S. A multilocus microsatellite data set of 836 individuals from 27 populations of T. infestans, from all its range of distribution in Argentina, was analyzed. Our results favor the hypothesis of two independent migration events of colonization in Argentina and secondary contacts. The majority of the populations of the western provinces of Catamarca, La Rioja, San Juan and the west of Cordoba province, had almost no shared ancestry with the rest of the populations analyzed. Probably those populations, belonging to localities close to the Andean region, could have been established by the dispersal line of T. infestans that would have arrived to Argentina through the Andes, whereas most of the rest of the populations analyzed may have derived from the dispersal line of T. infestans in non-Andean lowlands. Among them, those from the provinces of Formosa, Chaco, Santiago del Estero and Santa Fe shared different percentages of ancestry and presented lower degree of genetic differentiation. The migratory movement linked to regional economies and possibly associated with passive dispersal, would allow a higher genetic exchange among these populations of T. infestans. This study, using microsatellite markers, provides a new approach for evaluating the validity of the different hypotheses concerning the evolutionary history of this species. Two major lineages of T. infestans, an Andean and non-Andean, are suggested.     

Domestic host availability modifies human-triatomine contact and host shifts of the Chagas disease vector Triatoma infestans in the humid Argentine Chaco

Domestic animals may affect human-vector contact and parasite transmission rates. We investigated the relationships between host-feeding choices, site-specific host availability, bug nutritional status, stage and abundance of Triatoma infestans Klug (Heteroptera: Reduviidae) in rural houses of Pampa del Indio during spring. We identified the bloodmeal sources of 865 triatomines collected in 70 sites from four main ecotopes. The main sources in domiciles were human (65.9%), chicken (23.4%) and dog (22.4%); dog (64.4%, 35.3%) and chicken (33.1%, 75.4%) in kitchens and storerooms, respectively; and chicken (94.7%) in chicken coops. Using random-intercept logistic regression clustered by domicile, the fraction of human-fed triatomines strongly decreased with increasing proportions of chicken- and dog-fed bugs, dropping from 96.4% when no chicken or dog slept indoors at night to 59.4% when both did. The fraction of dog-fed bugs significantly decreased with increasing human and chicken blood indices, and marginally increased with an indoor-resting dog. Mixed blood meals occurred 3.62 times more often when a chicken or a dog slept indoors. Host blood source did not affect mean body weight adjusted for body length and bug stage. Indoor-resting chickens and dogs greatly modified human-bug contact rates, and may be targeted with long-lasting systemic insecticides to suppress infestation.     

Ecological Connectivity of Trypanosoma cruzi Reservoirs and Triatoma pallidipennis Hosts in an Anthropogenic Landscape with Endemic Chagas Disease

Traditional methods for Chagas disease prevention are targeted at domestic vector reduction, as well as control of transfusion and maternal-fetal transmission. Population connectivity of Trypanosoma cruzi-infected vectors and hosts, among sylvatic, ecotone and domestic habitats could jeopardize targeted efforts to reduce human exposure. This connectivity was evaluated in a Mexican community with reports of high vector infestation, human infection, and Chagas disease, surrounded by agricultural and natural areas. We surveyed bats, rodents, and triatomines in dry and rainy seasons in three adjacent habitats (domestic, ecotone, sylvatic), and measured T. cruzi prevalence, and host feeding sources of triatomines. Of 12 bat and 7 rodent species, no bat tested positive for T. cruzi, but all rodent species tested positive in at least one season or habitat. Highest T. cruzi infection prevalence was found in the rodents, Baiomys musculus and Neotoma mexicana. In general, parasite prevalence was not related to habitat or season, although the sylvatic habitat had higher infection prevalence than by chance, during the dry season. Wild and domestic mammals were identified as bloodmeals of T. pallidipennis, with 9% of individuals having mixed human (4.8% single human) and other mammal species in bloodmeals, especially in the dry season; these vectors tested >50% positive for T. cruzi. Overall, ecological connectivity is broad across this matrix, based on high rodent community similarity, vector and T. cruzi presence. Cost-effective T. cruzi, vector control strategies and Chagas disease transmission prevention will need to consider continuous potential for parasite movement over the entire landscape. This study provides clear evidence that these strategies will need to include reservoir/host species in at least ecotones, in addition to domestic habitats.     

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.     

Genetic transformation of a Corynebacterial symbiont from the Chagas disease vector Triatoma infestans

Insect-borne diseases have experienced a troubling resurgence in recent years. Emergence of resistance to pesticides greatly hampers control efforts. Paratransgenesis, or the genetic transformation of bacterial symbionts of disease vectors, is an alternative to traditional approaches. Previously, we developed paratransgenic lines of Rhodnius prolixus, a vector of Chagas disease in Central America. Here, we report identification of a Corynebacterial species as a symbiont of Triatoma infestans, a leading vector of Chagas disease in South America. We have modified this bacterium to produce an immunologically active single chain antibody fragment, termed rDB3. This study establishes the basis for generating paratransgenic T. infestans as a strategy for control of Chagas disease.     

Key Source Habitats and Potential Dispersal of Triatoma infestans Populations in Northwestern Argentina: Implications for Vector Control

Background

Triatoma infestans —the principal vector of the infection that causes Chagas disease— defies elimination efforts in the Gran Chaco region. This study identifies the types of human-made or -used structures that are key sources of these bugs in the initial stages of house reinfestation after an insecticide spraying campaign.

Methodology and Principal Findings

We measured demographic and blood-feeding parameters at two geographic scales in 11 rural communities in Figueroa, northwest Argentina. Of 1,297 sites searched in spring, 279 (21.5%) were infested. Bug abundance per site and female fecundity differed significantly among habitat types (ecotopes) and were highly aggregated. Domiciles (human sleeping quarters) had maximum infestation prevalence (38.7%), human-feeding bugs and total egg production, with submaximal values for other demographic and blood-feeding attributes. Taken collectively peridomestic sites were three times more often infested than domiciles. Chicken coops had greater bug abundance, blood-feeding rates, engorgement status, and female fecundity than pig and goat corrals. The host-feeding patterns were spatially structured yet there was strong evidence of active dispersal of late-stage bugs between ecotopes. Two flight indices predicted that female fliers were more likely to originate from kitchens and domiciles, rejecting our initial hypothesis that goat and pig corrals would dominate.

Conclusions and Significance

Chicken coops and domiciles were key source habitats fueling rapid house reinfestation. Focusing control efforts on ecotopes with human-fed bugs (domiciles, storerooms, goat corrals) would neither eliminate the substantial contributions to bug population growth from kitchens, chicken coops, and pig corrals nor stop dispersal of adult female bugs from kitchens. Rather, comprehensive control of the linked network of ecotopes is required to prevent feeding on humans, bug population growth, and bug dispersal simultaneously. Our study illustrates a demographic approach that may be applied to other regions and triatomine species for the design of innovative, improved vector control strategies.     

Reproductive and developmental costs of deltamethrin resistance in the Chagas disease vector Triatoma infestans

Effective chemical control relies on reducing vector population size. However, insecticide selection pressure is often associated with the development of resistant populations that reduce control success. In treated areas, these resistant individuals present an adaptive advantage due to enhanced survival. Resistance can also lead to negative effects when the insecticide pressure ceases. In this study, the biological effects of deltamethrin resistance were assessed in the Chagas disease vector Triatoma infestans. The length of each developmental stage and complete life cycle, mating rate, and fecundity were evaluated. Susceptible and resistant insects presented similar mating rates. A reproductive cost of resistance was expressed as a lower fecundity in the resistant colony. Developmental costs in the resistant colony were in the form of a shortening of the second and third nymph stage duration and an extension of the fifth stage. A maternal effect of deltamethrin resistance is suggested as these effects were identified in resistant females and their progeny independently of the mated male's deltamethrin response. Our results suggest the presence of pleiotropic effects of deltamethrin resistance. Possible associations of these characters to other traits such as developmental delays and behavioral resistance are discussed.     

Dispersive flight by Triatoma infestans under natural climatic conditions in Argentina

Male and female T. infestans were released on two consecutive nights in the salinas of Cordoba Province, Argentina, when air temperatures during the flying period averaged 28.5 degrees C. 136 males (43% of the total released) and 170 females (57%) flew on the first night, and 6 males (18%) and 7 females (27%) on the second. Of these, we recovered 23 males and 14 females within 100 m, and a further female within 200 m, all from the first night's release. The remainder appear to have flown more than 100 m and possibly more than 200 m. In support of this conclusion 4 male and 5 female bugs were recovered in an abandoned brick house 550 m from the release point. Since the proportion of bugs apparently flying more than 200 m is considerably greater than that deduced from previous experiments at lower temperatures (Lehane & Schofield, 1981) it is possible that temperature influences not only the proportion of bugs flying but also the distance flown. Flight appears to be affected by wind speed. On the first night, when there was negligible wind, 43% of male and 57% of female bugs flew. On the second night, with winds gusting at 4-5 m/s, only 18% of the male and 27% of the female bugs flew.     

Spatial pattern of reinfestation by Triatoma infestans in Chancanif, Argentina.

We examined the environmental correlates and the spatial pattern of infestation by Triatoma infestans, a vector of Chagas disease, in a rural area of Argentina five years following an insecticidal campaign. Patterns of infestation were identified in an entomological survey, as mapped with high-resolution satellite imagery and analyzed in a geographic information system. Logistic regression was used to relate infestation to observed household characteristics as well as the location and density of households. Location was the most significant predictor of infestation for domiciles. For peridomestic structures surrounding the domiciles, the combination of location and the presence/absence of goat pens was most significant. In considering any infestation, whether domiciliary or peridomestic, the combination of location, presence/absence of animal pens, and the type of household construction were found to be most significant. Using these statistical relationships to back-classify the field data resulted in accuracies between 85% and 87%. A map of infestation probability for the town of Chancaní was developed from the logistic regression.     

Mitochondrial 16S DNA variation in populations of Triatoma infestans from Argentina

Abstract.  Variation in the mtDNA 16S ribosomal RNA gene in populations of Triatoma infestans (Klug) was surveyed. DNA sequence comparisons yielded 18 haplotypes among 130 individuals from 16 localities that represent a large proportion of the range of T. infestans in Argentina. The most common genotype in all populations was found in 76.9% of individuals and two other haplotypes were shared among different populations. The remaining 15 haplotypes were present exclusively in one of the populations, suggesting currently low levels of genetic exchange. Analysis of mtDNA 16S sequences uncovered substantial genetic variation among T. infestans populations. Haplotype and nucleotide diversities varied among populations, from 0% to 0.84% and 0% to 0.29%, respectively. It appears that this locus has a low mutation rate. Uncorrected pairwise differences of T. infestans haplotypes ranged from 0% to 1.2%. The molecular phylogeny supported the monophyly of T. infestans haplotypes and clustered two different pairs of haplotypes with a moderate degree of bootstrap support (∼ 60%). Mitochondrial DNA phylogeographic differentiation was not evident, suggesting a recent rapid spread of the species. Analysis of molecular variance showed hierarchical structure in the data. Considerably less variation was found among T. infestans populations from the northwest and northeast regions than among those belonging to the central area. Such a lack of variation may be indicative of one or more past population bottlenecks.