Maria Jesus Ramirez-Sierra

Re-infestation of houses by Triatoma dimidiata after intra-domicile insecticide application in the Yucatán peninsula, Mexico.

In most countries, Chagas disease transmission control remains based on domestic insecticide application. We thus evaluated the efficacy of intra-domicile cyfluthrin spraying for the control of Triatoma dimidiata, the only Chagas disease vector in the Yucatán peninsula, Mexico, and monitored potential re-infestation every 15 days for up to 9 months. We found that there was a re-infestation of houses by adult bugs starting 4 months after insecticide application, possibly from sylvatic/peridomicile areas. This points out the need to take into account the potential dispersal of sylvatic/peridomestic adult bugs into the domiciles as well as continuity action for an effective vector control.

Patterns of house infestation dynamics by non‐domiciliated Triatoma dimidiata reveal a spatial gradient of infestation in rural villages and potential insect manipulation by Trypanosoma cruzi

Objective  Chagas disease is a major vector‐borne parasitic disease in Latin America, primarily transmitted to humans by triatomine vectors. Non‐domiciliated triatomine species such as Triatoma dimidiata in the Yucatan peninsula, Mexico, can transiently invade houses and are emerging as a major challenge to control Trypanosoma cruzi transmission to humans. We analyzed the spatio‐temporal spreading of house infestation by T. dimidiata in four rural villages.

Public Street Lights Increase House Infestation by the Chagas Disease Vector Triatoma dimidiata

Triatoma dimidiata is one of the primary vectors of Chagas disease. We previously documented the spatio-temporal infestation of houses by this species in the Yucatan peninsula, Mexico, and found that non-domiciliated triatomines were specifically attracted to houses. However, the factors mediating this attraction remained unclear. Artificial light has been known for a long time to attract many insect species, and therefore may contribute to the spread of different vector-borne diseases. Also, based on the collection of different species of triatomines with light traps, several authors have suggested that light might attract triatomines to houses, but the role of artificial light in house infestation has never been clearly demonstrated and quantified. Here we performed a spatial analysis of house infestation pattern by T. dimidiata in relation to the distribution of artificial light sources in three different villages from the Yucatan peninsula, Mexico. In all three villages, infested houses were significantly closer to public street light sources than non-infested houses (18.0±0.6 vs 22.6±0.4 m), and street lights rather than domestic lights were associated with house infestation. Accordingly, houses closer to a public street lights were 1.64 times more likely to be infested than houses further away (OR, CI95% 1.23–2.18). Behavioral experiments using a dual-choice chamber further confirmed that adult male and females were attracted to white light during their nocturnal activity. Attraction was also dependent on light color and decreased with increasing wavelength. While public lighting is usually associated with increased development, these data clearly show that it also directly contributes to house infestation by non-domiciliated T. dimidiata.

Identification of a large hybrid zone between sympatric sibling species of Triatoma dimidiata in the Yucatan peninsula, Mexico, and its epidemiological importance.

Triatoma dimidiata is one of the major Chagas disease vectors, with an extensive diversity in its morphology, habitat, and level of domiciliation. Molecular studies based on the internal transcribed spacer 2 (ITS-2) have subdivided this species into four potential taxonomic groups. Using both ITS-2 and cytochrome B markers, we confirmed the sibling species status of ITS-2 Group 3 and detected an apparent sympatry of ITS-2 Groups 2 and 3 in the Yucatan peninsula, Mexico. Here we examine the geographic distribution of T. dimidiata ITS-2 genotypes in the region and compare their egg production and Trypanosoma cruzi infection rates, as indicators of biological differences between groups. PCR genotyping of large natural populations showed an extensive sympatry of Groups 2 and 3 in most of the peninsula, often within the same house. We also detected a large proportion of individuals displaying ITS-2 sequences from both Groups 2 and 3, suggesting hybridization. Analysis of ITS-2 genotype frequencies indicated a strong departure from Hardy-Weinberg equilibrium in female hybrids, but not in males, due to a large heterozygote deficit. These results suggest random mating between ITS-2 Groups 2 and 3 combined with reduced viability and/or survival in female hybrids. This and other factors may allow for the maintenance of distinct ITS-2 Groups 2 and 3 populations despite high hybrid frequencies. Importantly, T. cruzi infection was much higher in hybrids compared to ITS-2 Groups 2 and 3 individuals, but all three genotypes appeared to seasonally infest houses in a similar manner in the region. These findings warrant further studies on T. dimidiata taxonomy and its epidemiologic implications.

Eco-Bio-Social Determinants for House Infestation by Non-domiciliated Triatoma dimidiata in the Yucatan Peninsula, Mexico

Background Chagas disease is a vector-borne disease of major importance in the Americas. Disease prevention is mostly limited to vector control. Integrated interventions targeting ecological, biological and social determinants of vector-borne diseases are increasingly used for improved control. Methodology/principal findings We investigated key factors associated with transient house infestation by T. dimidiata in rural villages in Yucatan, Mexico, using a mixed modeling approach based on initial null-hypothesis testing followed by multimodel inference and averaging on data from 308 houses from three villages. We found that the presence of dogs, chickens and potential refuges, such as rock piles, in the peridomicile as well as the proximity of houses to vegetation at the periphery of the village and to public light sources are major risk factors for infestation. These factors explain most of the intra-village variations in infestation. Conclusions/significance These results underline a process of infestation distinct from that of domiciliated triatomines and may be used for risk stratification of houses for both vector surveillance and control. Combined integrated vector interventions, informed by an Ecohealth perspective, should aim at targeting several of these factors to effectively reduce infestation and provide sustainable vector control.

Effect of a combination DNA vaccine for the prevention and therapy of Trypanosoma cruzi infection in mice: role of CD4+ and CD8+ T cells.

Chagas disease is a major public health problem, with about 10 million infected people, and DNA vaccines are a promising alternative for the control of Trypanosoma cruzi, the causing agent of the disease. We tested here a new DNA vaccine encoding a combination of two leading parasite antigens, TSA-1 and Tc24, for the prevention and therapy of T. cruzi infection. Immunized Balb/c mice challenged by T. cruzi presented a significantly lower parasitemia and inflammatory cell density in the heart compared to control mice. Similarly, the therapeutic administration of the DNA vaccine was able to significantly reduce the parasitemia and inflammatory reaction in acutely infected Balb/c and C57BL/6 mice, and reduced cardiac tissue inflammation in chronically infected ICR mice. Therapeutic vaccination induced a marked increase in parasite-specific IFNγ producing CD4(+) and CD8(+) T cells in the spleen as well as an increase in CD4(+) and CD8(+) T cells in the infected cardiac tissue. In addition, some effect of the DNA vaccine could still be observed in CD4-knockout C57BL/6 mice, which presented a lower parasitemia and inflammatory cell density, but not in CD8-deficient mice, in which the vaccine had no effect. These results indicate that the activation of CD8(+) T cells plays a major role in the control of the infection by the therapeutic DNA vaccine, and to a somewhat lesser extent CD4(+) T cells. This observation opens interesting perspectives for the potentiation of this DNA vaccine candidate by including additional CD8(+) T cell antigens/epitopes in future vaccine formulations.

Mining the Leishmania genome for novel antigens and vaccine candidates

Leishmaniasis is a neglected disease with an estimated 12 million infected people. The recent completion of the sequencing of the Leishmania major genome has opened opportunities for the identification of targets for vaccine development. We present here the first attempt at identifying novel vaccine candidates by whole genome analysis. We predicted CD8+ T cell epitopes from the L. major proteome and validated in vivo in mice the immunogenicity of some of the best predicted epitopes. Consensus epitope predictions from 8272 annotated protein sequences with 5–8 different algorithms allowed the identification of 78 class I CD8+ epitopes. BALB/c mice were immunized with 26 synthetic peptides corresponding to the most likely epitopes. Fourteen (54%) resulted immunogenic, with eight being strong inducers of T cell IFNγ production. None of the proteins from which the epitopes are derived are differentially expressed, only two may be surface proteins, eight have putative enzymatic, and metabolic activities. These epitopes and proteins represent new antigen candidates for further studies. While pathogen genomes have not yet delivered their full promise in terms of human health applications, our study opens the way for extensive genome mining for antigen identification and vaccine development against Leishmania and other pathogens.

An innovative ecohealth intervention for Chagas disease vector control in Yucatan, Mexico

Background Non-domiciliated (intrusive) triatomine vectors remain a challenge for the sustainability of Chagas disease vector control as these triatomines are able to transiently (re-)infest houses. One of the best-characterized examples is Triatoma dimidiata from the Yucatan peninsula, Mexico, where adult insects seasonally infest houses between March and July. Methods We focused our study on three rural villages in the state of Yucatan, Mexico, in which we performed a situation analysis as a first step before the implementation of an ecohealth (ecosystem approach to health) vector control intervention. Results The identification of the key determinants affecting the transient invasion of human dwellings by T. dimidiata was performed by exploring associations between bug presence and qualitative and quantitative variables describing the ecological, biological and social context of the communities. We then used a participatory action research approach for implementation and evaluation of a control strategy based on window insect screens to reduce house infestation by T. dimidiata. Conclusions This ecohealth approach may represent a valuable alternative to vertically-organized insecticide spraying. Further evaluation may confirm that it is sustainable and provides effective control (in the sense of limiting infestation of human dwellings and vector/human contacts) of intrusive triatomines in the region.

Effects of genetic factors and infection status on wing morphology of Triatoma dimidiata species complex in the Yucatán peninsula, Mexico.

Triatoma dimidiata is one of the main vectors of Chagas disease, and it has been shown to be a species complex. In the Yucatán peninsula, Mexico, vector populations are non-domiciliated, and the transmission of Trypanosoma cruzi thus critically relies on vector dispersal. This leads us to study the morphologic variations in T. dimidiata wings with respect to genetic factors (sex and genotype at the ITS-2 locus), geographic location, and T. cruzi-infection status. Females were found to have larger and more symmetrical wings than males. Wing shape was influenced by ITS-2 genotypes, although differences are unlikely sufficient to allow taxonomic discrimination of the sibling species. Hybrids were shown to have similar fluctuating asymmetries in wing size and shape as parental species, but the level of asymmetry in shape varied slightly between villages. The two later findings are consistent with a high level of gene flow between parental species, and the high dispersal potential of these non-domiciliated vectors. More surprisingly, individuals infected with T. cruzi were found to have larger wings than non-infected ones. This effect, which was consistently observed across sexes, genotypes and villages, is likely to be due to a direct impact of T. cruzi on insect development. Sex and infection status are thus likely to be key factors influencing vector dispersal with important impacts on disease transmission, since dispersal directly controls the domestic abundance of these vectors. These aspects should be investigated further to fully capture the ecology and evolution of Chagas disease transmission by non-domiciliated vectors.

Comparative Field Trial of Alternative Vector Control Strategies for Non-Domiciliated Triatoma dimidiata

Chagas disease is a major vector-borne disease, and regional initiatives based on insecticide spraying have successfully controlled domiciliated vectors in many regions. Non-domiciliated vectors remain responsible for a significant transmission risk, and their control is a challenge. We performed a proof-of-concept field trial to test alternative strategies in rural Yucatan, Mexico. Follow-up of house infestation for two seasons following the interventions confirmed that insecticide spraying should be performed annually for the effective control of Triatoma dimidiata; however, it also confirmed that insect screens or long-lasting impregnated curtains may represent good alternative strategies for the sustained control of these vectors. Ecosystemic peridomicile management would be an excellent complementary strategy to improve the cost-effectiveness of interventions. Because these strategies would also be effective against other vector-borne diseases, such as malaria or dengue, they could be integrated within a multi-disease control program.