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Transmission risk of Chagas disease has been associated with human-vector contacts and triatomines colonizing dwellings, but alternative scenarios, independent of domestic colonization, are poorly documented. In the present work, we estimated the frequency of human blood meals in triatomines from domicile, peridomicile, and sylvatic environments in two endemic regions in Ecuador. Blood meal origins were identified by sequencing a cytb gene fragment. Human blood meals were detected in 42% of the triatomines among 416 analyzed, including 48% of sylvatic triatomines (both adults and nymphs). In triatomines from domicile and peridomicile, Trypanosoma cruzi infection rate was > 20%, and reached 48% in sylvatic triatomines. Human is a common source of blood for triatomines whether they live in or near dwellings in both regions, and the high rate of T. cruzi infection represents an important risk of transmission of Chagas disease. Consequently, control strategies should also take into account possible nondomestic transmission.

Introduction: Control of triatomine infestation is a key strategy for the prevention of Chagas disease (CD). To promote this strategy, it is important to know which antecedents to behavioral change are the best to emphasize when promoting prevention., Objective: The aim of this study was to determine predictors for intention to prevent home infestation based on the Health Belief Model (HBM), a commonly used health intervention planning theory., Materials & Methods: A cross-sectional study was conducted with 112 heads of household in six communities with endemic and high rates of triatomine infestation in Loja province, Ecuador. The data was collected by a questionnaire including perceived severity, susceptibility, benefits to action, barriers to action, and self-efficacy. These data were also used to predict actual infestation of homes., Results: Community members reported strong intentions to prevent home infestation. HBM constructs predicted about 14% of the observed variance in intentions. Perceived susceptibility and severity did not predict behavioral intention well; perceived barriers to small-scale action that reduce likelihood of infestation and self-efficacy in participating in surveillance systems did. Self-efficacy and perception of barriers were equally powerful predictors. The HBM constructs, however, did not predict well actual infestation., Conclusion: The findings supported the HBM as a way to predict intentions to prevent infestation of the home by triatomine bugs. The findings highlight that messages emphasizing self-efficacy in participating in surveillance systems and overcoming barriers to small-scale action that reduce likelihood of infestation, rather than a focus on risk, should be central messages when designing and implementing educational interventions for CD. The gap between behavioral intention and actual infestation reveals the need to assess home practices and their actual efficacy to fully enact and apply the HBM., Competing Interests: The authors have declared that no competing interests exist.

Background: Eggs have epidemiological and taxonomic importance in the subfamily Triatominae, which contains Chagas disease vectors. The metric properties (size and shape) of eggs are useful for distinguishing between close species, or different geographical populations of the same species., Methods: We examined the effects of egg viability on its metric properties, and the possible consequences on species recognition. Four species were considered: Panstrongylus chinai, P. howardi and Triatoma carrioni (tribe Triatomini), and Rhodnius ecuadoriensis (tribe Rhodniini). Digitization was performed on pictures taken when the viability of the egg could not clearly be predicted by visual inspection. We then followed development to separate viable from non-viable eggs, and the metric changes associated with viability status of the eggs were tested for species discrimination (interspecific difference)., Results: The shape of the complete contour of the egg provided satisfactory species classification (95% of correct assignments, on average), with improved scores (98%) when discarding non-viable eggs from the comparisons. Using only non-viable eggs, the scores dropped to 90%. The morphometric differences between viable and non-viable eggs were also explored (intraspecific comparison). A constant metric change observed was a larger variance of size and shape in non-viable eggs. For all species, larger eggs, or eggs with larger operculum, were more frequently non-viable. However, these differences did not allow for an accurate prediction regarding egg viability., Conclusions: The strong taxonomic signal present in egg morphology was affected by the level of viability of the eggs. The metric properties as modified in non-viable eggs presented some general trends which could suggest the existence of an optimum phenotype for size and for shape. Globally, viable eggs tended to have intermediate or small sizes, and presented a less globular shape in the Triatomini, or a relatively wider neck in Rhodnius ecuadoriensis.

Background: Chagas disease (CD) is a life-threatening illness caused by the protozoan parasite Trypanosoma cruzi, which is transmitted by triatomine bugs. Triatomine bugs inhabit poorly constructed homes that create multiple hiding spots for the bugs. Modifying the actual structure of a home, along with the homeowners' practices, can reduce triatomine infestation. This research was designed to collect culturally-relevant information to develop a health campaign to decrease risk of CD transmission by promoting home maintenance and better hygiene in rural communities of southern Ecuador., Methods and Main Findings: The Health Belief Model (HBM) guided focus group discussions and the interpretation of the results. Four focus groups ranging from 4 to 10 participants were conducted between May and June 2014 in three communities of Loja province in Southern Ecuador. A thematic analysis was used to identify within the data related to perceptions of susceptibility, severity, benefits, barriers and self-efficacy related to CD and its prevention. The results provide clear guidance for the development of Chagas-prevention messages., Conclusion: Data obtained emphasize the importance of standardizing messages presented to the communities for CD prevention. Messages should provide more information on the protective nature of the behaviors promoted for CD prevention; overcoming barriers such as cost and convenience, and build on facilitating factors, including community members' interest on quality of life, protection of their families, and relationship with the land., Competing Interests: The authors have declared that no competing interests exist.

Background: Egg morphometrics in the Triatominae has proved to be informative for distinguishing tribes or genera, and has been based generally on traditional morphometrics. However, more resolution is required, allowing species or even population recognition, because the presence of eggs in the domicile could be related to the species ability to colonize human dwellings, suggesting its importance as a vector., Results: We explored the resolution of modern morphometric methods to distinguish not only tribes and genera, but also species or geographic populations in some important Triatominae. Four species were considered, representing two tribes and three genera: Panstrongylus chinai and P. howardi, Triatoma carrioni and Rhodnius ecuadoriensis. Within R. ecuadoriensis, two geographical populations of Ecuador were compared. For these comparisons, we selected the most suitable day of egg development, as well as the possible best position of the egg for data capture. The shape of the eggs in the Triatominae does not offer true anatomical landmarks as the ones used in landmark-based morphometrics, except for the egg cap, especially in eggs with an evident "neck", such as those of the Rhodniini. To capture the operculum shape variation, we used the landmark- and semilandmark-based method. The results obtained from the metric properties of the operculum were compared with the ones provided by the simple contour of the whole egg, as analyzed by the Elliptic Fourier Analysis. Clear differences could be disclosed between the genera, between the species - among which two very close species (P. chinai and P. howardi), as well as between two allopatric, conspecific populations. The whole egg contour (including the operculum) produced reclassification scores much more satisfactory than the ones obtained using the operculum only., Conclusions: We propose the outline-based approach as the most convenient characterization tool to identify unknown eggs at the species or population levels.

Chagas disease is caused by Trypanosoma cruzi, which is mainly transmitted by the faeces of triatomine insects that find favourable environments in poorly constructed houses. Previous studies have documented persistent triatomine infestation in houses in the province of Loja in southern Ecuador despite repeated insecticide and educational interventions. We aim to develop a sustainable strategy for the interruption of Chagas disease transmission by promoting living environments that are designed to prevent colonisation of rural houses by triatomines. This study used positive deviance to inform the design of an anti-triatomine prototype house by identifying knowledge, attitudes and practices used by families that have remained triatomine-free (2010-2012). Positive deviants reported practices that included maintenance of structural elements of the house, fumigation of dwellings and animal shelters, sweeping with "insect repellent" plants and relocation of domestic animals away from the house, among others. Participants favoured construction materials that do not drastically differ from those currently used (adobe walls and tile roofs). They also expressed their belief in a clear connection between a clean house and health. The family's economic dynamics affect space use and must be considered in the prototype's design. Overall, the results indicate a positive climate for the introduction of housing improvements as a protective measure against Chagas disease in this region.

Background: Molecular epidemiology at the community level has an important guiding role in zoonotic disease control programmes where genetic markers are suitably variable to unravel the dynamics of local transmission. We evaluated the molecular diversity of Trypanosoma cruzi, the etiological agent of Chagas disease, in southern Ecuador (Loja Province). This kinetoplastid parasite has traditionally been a paradigm for clonal population structure in pathogenic organisms. However, the presence of naturally occurring hybrids, mitochondrial introgression, and evidence of genetic exchange in the laboratory question this dogma., Methodology/principal Findings: Eighty-one parasite isolates from domiciliary, peridomiciliary, and sylvatic triatomines and mammals were genotyped across 10 variable microsatellite loci. Two discrete parasite populations were defined: one predominantly composed of isolates from domestic and peridomestic foci, and another predominantly composed of isolates from sylvatic foci. Spatial genetic variation was absent from the former, suggesting rapid parasite dispersal across our study area. Furthermore, linkage equilibrium between loci, Hardy-Weinberg allele frequencies at individual loci, and a lack of repeated genotypes are indicative of frequent genetic exchange among individuals in the domestic/peridomestic population., Conclusions/significance: These data represent novel population-level evidence of an extant capacity for sex among natural cycles of T. cruzi transmission. As such they have dramatic implications for our understanding of the fundamental genetics of this parasite. Our data also elucidate local disease transmission, whereby passive anthropogenic domestic mammal and triatomine dispersal across our study area is likely to account for the rapid domestic/peridomestic spread of the parasite. Finally we discuss how this, and the observed subdivision between sympatric sylvatic and domestic/peridomestic foci, can inform efforts at Chagas disease control in Ecuador.

Rhodnius ecuadoriensis is considered the second most important vector of Chagas disease in Ecuador. It is distributed across six of the 24 provinces and occupies intradomiciliary, peridomiciliary and sylvatic habitats. This study was conducted in six communities within the coastal province of Guayas. Triatomine searches were conducted in domestic and peridomestic habitats and bird nests using manual searches, live-bait traps and sensor boxes. Synantrhopic mammals were captured in the domestic and peridomestic habitats. Household searches (n = 429) and randomly placed sensor boxes (n = 360) produced no live triatomine adults or nymphs. In contrast, eight nymphs were found in two out of six searched Campylorhynchus fasciatus (Troglodytidae) nests. Finally, Trypanosoma cruzi DNA was amplified from the blood of 10% of the 115 examined mammals. Environmental changes in land use (intensive rice farming), mosquito control interventions and lack of intradomestic adaptation are suggested among the possible reasons for the lack of domestic triatomine colonies.