RESUMO
Triatoma dimidiata is the main vector of Chagas disease in southern Mexico, Central America and northern South America. As a native vector, it moves readily among domestic, peridomestic and sylvatic environments, making it difficult to control only using insecticide as this requires regular application, and re-infestation frequently occurs. Other social innovation alternatives such as those based on Ecohealth principles can be used to tackle the dynamics of the disease in an integral way. We asked whether an Ecohealth intervention, implemented beginning in 2001 in a highly infested village, 41.8%, in southeastern Guatemala, was sustainable in the long term. This intervention included initial insecticide treatments, followed by making low-cost house improvements to eliminate transmission risk factors such as repairing cracked walls, covering dirt floors with a cement-like substance and moving domestic animals outside. We assessed the long-term sustainability through entomological and house condition surveys, as well as an analysis of community satisfaction. We found over a 19-year period, infestation with T. dimidiata was reduced to 2.2% and maintained at a level below the level (8%) where vector transmission is unlikely. This long-term maintenance of low infestation coincided with a large proportion of villagers (88.6%) improving their houses and completing other aspects of the Ecohealth approach to maintain the village at low risk for Chagas transmission. There was unanimous satisfaction among the villagers with their houses, following improvements using the Ecohealth method, which likely played a role in the long-term persistence of the modifications. Although the infestation has remained low, 11 years following the last intervention and as the population grew there has been an increase in the proportion of "at-risk" houses, to 33%, pointing out the necessity of maintaining vigilance. The Ecohealth approach is a low-cost, sustainable approach for the long-term control of vector-borne Chagas disease. We recommend this approach including ongoing community monitoring and institutional response for the long-term, integrated control of Chagas disease.
Assuntos
Doença de Chagas , Inseticidas , Triatoma , Animais , Doença de Chagas/prevenção & controle , Guatemala/epidemiologia , Habitação , Controle de Insetos/métodos , Insetos Vetores/fisiologia , Triatoma/fisiologiaRESUMO
More than 100 years since the first description of Chagas Disease and with over 29,000 new cases annually due to vector transmission (in 2010), American Trypanosomiasis remains a Neglected Tropical Disease (NTD). This study presents the most comprehensive Trypanosoma cruzi sampling in terms of geographic locations and triatomine species analyzed to date and includes both nuclear and mitochondrial genomes. This addresses the gap of information from North and Central America. We incorporate new and previously published DNA sequence data from two mitochondrial genes, Cytochrome oxidase II (COII) and NADH dehydrogenase subunit 1 (ND1). These T. cruzi samples were collected over a broad geographic range including 111 parasite DNA samples extracted from triatomines newly collected across North and Central America, all of which were infected with T. cruzi in their natural environment. In addition, we present parasite reduced representation (Restriction site Associated DNA markers, RAD-tag) genomic nuclear data combined with the mitochondrial gene sequences for a subset of the triatomines (27 specimens) collected from Guatemala and El Salvador. Our mitochondrial phylogenetic reconstruction revealed two of the major mitochondrial lineages circulating across North and Central America, as well as the first ever mitochondrial data for TcBat from a triatomine collected in Central America. Our data also show that within mtTcIII, North and Central America represent an independent, distinct clade from South America, named here as mtTcIIINA-CA, geographically restricted to North and Central America. Lastly, the most frequent lineage detected across North and Central America, mtTcI, was also an independent, distinct clade from South America, noted as mtTcINA-CA. Furthermore, nuclear genome data based on Single Nucleotide Polymorphism (SNP) showed genetic structure of lineage TcI from specimens collected in Guatemala and El Salvador supporting the hypothesis that genetic diversity at a local scale has a geographical component. Our multiscale analysis contributes to the understanding of the independent and distinct evolution of T. cruzi lineages in North and Central America regions.
Assuntos
Doença de Chagas/parasitologia , Mitocôndrias/genética , Trypanosoma cruzi/classificação , Trypanosoma cruzi/isolamento & purificação , América Central , Complexo I de Transporte de Elétrons/genética , Complexo I de Transporte de Elétrons/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/genética , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Humanos , Mitocôndrias/metabolismo , Filogenia , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , América do Sul , Trypanosoma cruzi/genéticaRESUMO
Assays for parasite detection in insect vectors provide important information for disease control. American Trypanosomiasis (Chagas disease) is the most devastating vector-borne illness and the fourth most common in Central America behind HIV/AIDS and acute respiratory and diarrheal infections (Peterson et al., 2019). Under-detection of parasites is a general problem which may be influenced by parasite genetic variation; however, little is known about the genetic variation of the Chagas parasite, especially in this region. In this study we compared six assays for detecting the Chagas parasite, Trypanosoma cruzi: genomic reduced representation sequencing (here referred to as genotype-by-sequencing or GBS), two with conventional PCR (i.e., agarose gel detection), two with qPCR, and microscopy. Our results show that, compared to GBS genomic analysis, microscopy and PCR under-detected T. cruzi in vectors from Central America. Of 94 samples, 44% (50/94) were positive based on genomic analysis. The lowest detection, 9% (3/32) was in a subset assayed with microscopy. Four PCR assays, two with conventional PCR and two with qPCR showed intermediate levels of detection. Both qPCR tests and one conventional PCR test targeted the 195 bp repeat of satellite DNA while the fourth test targeted the 18S gene. Statistical analyses of the genomic and PCR results indicate that the PCR assays significantly under detect infections of Central American T. cruzi genotypes.
Assuntos
Doença de Chagas , Triatoma , Trypanosoma cruzi , Animais , América Central , Doença de Chagas/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real , Triatoma/genética , Trypanosoma cruzi/genéticaRESUMO
Chagas disease is a lethal, neglected tropical disease. Unfortunately, aggressive insecticide-spraying campaigns have not been able to eliminate domestic infestation of Triatoma dimidiata, the native vector in Guatemala. To target interventions toward houses most at risk of infestation, comprehensive socioeconomic and entomologic surveys were conducted in two towns in Jutiapa, Guatemala. Given the exhaustively large search space associated with combinations of risk factors, traditional statistics are limited in their ability to discover risk factor interactions. Two recently developed statistical evolutionary algorithms, specifically designed to accommodate risk factor interactions and heterogeneity, were applied to this large combinatorial search space and used in tandem to identify sets of risk factor combinations associated with infestation. The optimal model includes 10 risk factors in what is known as a third-order disjunctive normal form (i.e., infested households have chicken coops AND deteriorated bedroom walls OR an accumulation of objects AND dirt floors AND total number of occupants ≥ 5 AND years of electricity ≥ 5 OR poor hygienic condition ratings AND adobe walls AND deteriorated walls AND dogs). Houses with dirt floors and deteriorated walls have been reported previously as risk factors and align well with factors currently targeted by Ecohealth interventions to minimize infestation. However, the tandem evolutionary algorithms also identified two new socioeconomic risk factors (i.e., households having many occupants and years of electricity ≥ 5). Identifying key risk factors may help with the development of new Ecohealth interventions and/or reduce the survey time needed to identify houses most at risk.
Assuntos
Animais Domésticos , Doença de Chagas/epidemiologia , Materiais de Construção/estatística & dados numéricos , Abrigo para Animais , Habitação/estatística & dados numéricos , Insetos Vetores , Triatoma , Algoritmos , Animais , Doença de Chagas/transmissão , Galinhas , Cães , Instalação Elétrica/estatística & dados numéricos , Características da Família , Guatemala/epidemiologia , Humanos , Higiene , Controle de Insetos , Inseticidas , Piretrinas , Fatores de Risco , Comportamento de Redução do Risco , Fatores SocioeconômicosRESUMO
Chagas disease is caused by the protozoan parasite Trypanosoma cruzi and transmitted by triatomine insect vectors. In Guatemala, insecticide spraying is an integral part of management of the main vector, Triatoma dimidiata. Spraying typically has low efficacy, which may be due to incomplete elimination from infested houses, within-village dispersal, or influx from other villages or sylvan environments. To evaluate how these mechanisms contribute to reinfestation, we conducted a time-course analysis of T. dimidiata infestation, abundance and household genetic structure in two nearby villages in Jutiapa, Guatemala; houses in the first village were surveyed, treated with insecticide if infested and then re-surveyed at eight and 22â¯months following spraying, while the second village served as an untreated control to quantify changes associated with seasonal dispersal. Insects were genotyped at 2-3000 SNP loci for kinship and population genetic analyses. Insecticide application reduced overall infestation and abundance, while the untreated village was stable over time. Nevertheless, within two years 35.5% of treated houses were reinfested and genetic diversity had largely recovered. Insects collected from reinfested houses post-spraying were most closely related to pre-spray collections from the same house, suggesting that infestations had not been fully eliminated. Immigration by unrelated insects was also detected within a year of spraying; when it occurred, dispersal was primarily local from neighboring houses. Similar dispersal patterns were observed following the annual dispersal season in the untreated village, with high-infestation houses serving as sources for neighboring homes. Our findings suggest that the efficacy of pyrethroid application is rapidly diminished by both within-house breeding by survivors and annual cycles of among-house movement. Given these patterns, we conclude that house structural improvements, an integral part of the Ecohealth approach that makes houses refractory to vector colonization and persistence, are critical for long-term reduction of T. dimidiata infestation.
Assuntos
Resistência a Inseticidas , Inseticidas/farmacologia , Polimorfismo de Nucleotídeo Único , Piretrinas/farmacologia , Triatoma/crescimento & desenvolvimento , Animais , DNA/genética , Feminino , Técnicas de Genotipagem/métodos , Guatemala , Controle de Insetos , Masculino , Dinâmica Populacional , Triatoma/efeitos dos fármacos , Triatoma/genéticaRESUMO
OBJECTIVES: We tested a rapid and specific immunochromatographic assay (that detects human blood in forensic samples) to determine if human blood was present in triatomines and their fecal excreta. METHODS: We fed Triatoma rubida human blood (positive control) or mouse blood (negative control) and performed the assay on the abdominal contents and fecal excreta. Triatomine field specimens collected in and around human habitations and excreta were also tested. FINDINGS: The assay was positive in triatomines fed human blood (N = 5/5) and fecal excreta from bugs known to have ingested human blood (N = 5/5). Bugs feeding on mice (N = 15/15) and their fecal excreta (N = 8/8) were negative for human blood. Human blood was detected in 47% (N = 23/49) triatomines, representing six different species, collected in the field. MAIN CONCLUSIONS: The pilot study shows that this rapid and specific test may have applications in triatomine research. Further study is needed to determine the sensitivity of this assay compared to other well-established techniques, such as DNA- and proteomics-based methodologies and the assay's application in the field.
Assuntos
Sangue , Fezes/química , Imunoensaio/métodos , Triatominae , Animais , Doença de Chagas/transmissão , Humanos , Camundongos , Projetos Piloto , Padrões de Referência , Valores de Referência , Reprodutibilidade dos Testes , Fatores de TempoRESUMO
BACKGROUND DNA- and proteomics-based techniques are currently used to identify a triatomine human blood meal. These methods are time consuming, require access to laboratories with sophisticated equipment, and trained personnel. OBJECTIVES We tested a rapid and specific immunochromatographic assay (that detects human blood in forensic samples) to determine if human blood was present in triatomines and their fecal excreta. METHODS We fed Triatoma rubida human blood (positive control) or mouse blood (negative control) and performed the assay on the abdominal contents and fecal excreta. Triatomine field specimens collected in and around human habitations and excreta were also tested. FINDINGS The assay was positive in triatomines fed human blood (N = 5/5) and fecal excreta from bugs known to have ingested human blood (N = 5/5). Bugs feeding on mice (N = 15/15) and their fecal excreta (N = 8/8) were negative for human blood. Human blood was detected in 47% (N = 23/49) triatomines, representing six different species, collected in the field. MAIN CONCLUSIONS The pilot study shows that this rapid and specific test may have applications in triatomine research. Further study is needed to determine the sensitivity of this assay compared to other well-established techniques, such as DNA- and proteomics-based methodologies and the assay's application in the field.
Assuntos
Humanos , Imunoensaio , Cromatografia de Afinidade/métodos , Triatominae , Projetos PilotoRESUMO
Chagas disease, considered a neglected disease by the World Health Organization, is caused by the protozoan parasite Trypanosoma cruzi, and transmitted by >140 triatomine species across the Americas. In Central America, the main vector is Triatoma dimidiata, an opportunistic blood meal feeder inhabiting both domestic and sylvatic ecotopes. Given the diversity of interacting biological agents involved in the epidemiology of Chagas disease, having simultaneous information on the dynamics of the parasite, vector, the gut microbiome of the vector, and the blood meal source would facilitate identifying key biotic factors associated with the risk of T. cruzi transmission. In this study, we developed a RADseq-based analysis pipeline to study mixed-species DNA extracted from T. dimidiata abdomens. To evaluate the efficacy of the method across spatial scales, we used a nested spatial sampling design that spanned from individual villages within Guatemala to major biogeographic regions of Central America. Information from each biotic source was distinguished with bioinformatics tools and used to evaluate the prevalence of T. cruzi infection and predominant Discrete Typing Units (DTUs) in the region, the population genetic structure of T. dimidiata, gut microbial diversity, and the blood meal history. An average of 3.25 million reads per specimen were obtained, with approximately 1% assigned to the parasite, 20% to the vector, 11% to bacteria, and 4% to putative blood meals. Using a total of 6,405 T. cruzi SNPs, we detected nine infected vectors harboring two distinct DTUs: TcI and a second unidentified strain, possibly TcIV. Vector specimens were sufficiently variable for population genomic analyses, with a total of 25,710 T. dimidiata SNPs across all samples that were sufficient to detect geographic genetic structure at both local and regional scales. We observed a diverse microbiotic community, with significantly higher bacterial species richness in infected T. dimidiata abdomens than those that were not infected. Unifrac analysis suggests a common assemblage of bacteria associated with infection, which co-occurs with the typical gut microbial community derived from the local environment. We identified vertebrate blood meals from five T. dimidiata abdomens, including chicken, dog, duck and human; however, additional detection methods would be necessary to confidently identify blood meal sources from most specimens. Overall, our study shows this method is effective for simultaneously generating genetic data on vectors and their associated parasites, along with ecological information on feeding patterns and microbial interactions that may be followed up with complementary approaches such as PCR-based parasite detection, 18S eukaryotic and 16S bacterial barcoding.
Assuntos
DNA/genética , DNA/isolamento & purificação , Comportamento Alimentar , Microbioma Gastrointestinal , Triatoma/genética , Trypanosoma cruzi/isolamento & purificação , Animais , Archaea/genética , Archaea/isolamento & purificação , Bactérias/genética , Bactérias/isolamento & purificação , América Central , Análise por Conglomerados , Biologia Computacional , Fungos/genética , Fungos/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Nematoides/genética , Nematoides/isolamento & purificação , Filogenia , Análise de Sequência de DNA , Triatoma/microbiologia , Triatoma/parasitologia , Triatoma/fisiologia , Trypanosoma cruzi/genética , Vírus/genética , Vírus/isolamento & purificaçãoRESUMO
In this paper, Triatoma mopansp. n. is described based on five males and six females collected in the Rio Frio cave, Cayo District, Belize. This species is similar to Triatoma dimidiata (Latreille), but can be distinguished by characters found on the pronotum, legs, and abdomen. Geometric morphometry and phylogenetic comparisons are also provided. Presently, the species is known only from the type locality and is a potential Chagas vector.
RESUMO
To date, the phylogeny of Triatoma dimidiata sensu lato (s. l.) (Hemiptera: Reduviidae: Triatominae), the epidemiologically most important Chagas disease vector in Central America and a secondary vector in Mexico and northern South America, has only been investigated by one multi-copy nuclear gene (Internal Transcribed Spacer - 2) and a few mitochondrial genes. We examined 450 specimens sampled across most of its native range from Mexico to Ecuador using reduced representation next-generation sequencing encompassing over 16,000 single nucleotide polymorphisms (SNPs). Using a combined phylogenetic and species delimitation approach we uncovered two distinct species, as well as a well-defined third group that may contain multiple species. The findings are discussed with respect to possible drivers of diversification and the epidemiological importance of the distinct species and groups.
Assuntos
Variação Genética , Genoma , Triatoma/genética , Animais , América Central , Doença de Chagas/parasitologia , Doença de Chagas/patologia , DNA/química , DNA/isolamento & purificação , DNA/metabolismo , Genes Mitocondriais , Humanos , Insetos Vetores/genética , Filogenia , Polimorfismo de Nucleotídeo Único , Triatoma/classificação , Triatoma/parasitologia , Trypanosoma cruzi/fisiologiaRESUMO
Little is known about the strains of Trypanosoma cruzi circulating in Central America and specifically in the most important vector in this region, Triatoma dimidiata. Approximately six million people are infected with T. cruzi, the causative agent of Chagas disease, which has the greatest negative economic impact and is responsible for ~12,000 deaths annually in Latin America. By international consensus, strains of T. cruzi are divided into six monophyletic clades called discrete typing units (DTUs TcI-VI) and a seventh DTU first identified in bats called TcBat. TcI shows the greatest geographic range and diversity. Identifying strains present and diversity within these strains is important as different strains and their genotypes may cause different pathologies and may circulate in different localities and transmission cycles, thus impacting control efforts, treatment and vaccine development. To determine parasite strains present in T. dimidiata across its geographic range from Mexico to Colombia, we isolated abdominal DNA from T. dimidiata and determined which specimens were infected with T. cruzi by PCR. Strains from infected insects were determined by comparing the sequence of the 18S rDNA and the spliced-leader intergenic region to typed strains in GenBank. Two DTUs were found: 94% of infected T. dimidiata contained TcI and 6% contained TcIV. TcI exhibited high genetic diversity. Geographic structure of TcI haplotypes was evident by Principal Component and Median-Joining Network analyses as well as a significant result in the Mantel test, indicating isolation by distance. There was little evidence of association with TcI haplotypes and host/vector or ecotope. This study provides new information about the strains circulating in the most important Chagas vector in Central America and reveals considerable variability within TcI as well as geographic structuring at this large geographic scale. The lack of association with particular vectors/hosts or ecotopes suggests the parasites are moving among vectors/hosts and ecotopes therefore a comprehensive approach, such as the Ecohealth approach that makes houses refractory to the vectors will be needed to successfully halt transmission of Chagas disease.
Assuntos
Doença de Chagas/parasitologia , Variação Genética , Insetos Vetores/parasitologia , Triatoma/parasitologia , Trypanosoma cruzi/genética , Animais , Doença de Chagas/epidemiologia , Doença de Chagas/transmissão , Quirópteros/parasitologia , Colômbia/epidemiologia , Genótipo , Haplótipos , Humanos , México/epidemiologia , Filogenia , Trypanosoma cruzi/classificação , Trypanosoma cruzi/isolamento & purificação , Trypanosoma cruzi/fisiologiaRESUMO
The widespread and diverse Triatoma dimidiata is the kissing bug species most important for Chagas disease transmission in Central America and a secondary vector in Mexico and northern South America. Its diversity may contribute to different Chagas disease prevalence in different localities and has led to conflicting systematic hypotheses describing various populations as subspecies or cryptic species. To resolve these conflicting hypotheses, we sequenced a nuclear (internal transcribed spacer 2, ITS-2) and mitochondrial gene (cytochrome b) from an extensive sampling of T. dimidiata across its geographic range. We evaluated the congruence of ITS-2 and cyt b phylogenies and tested the support for the previously proposed subspecies (inferred from ITS-2) by: (1) overlaying the ITS-2 subspecies assignments on a cyt b tree and, (2) assessing the statistical support for a cyt b topology constrained by the subspecies hypothesis. Unconstrained phylogenies inferred from ITS-2 and cyt b are congruent and reveal three clades including two putative cryptic species in addition to T. dimidiata sensu stricto. Neither the cyt b phylogeny nor hypothesis testing support the proposed subspecies inferred from ITS-2. Additionally, the two cryptic species are supported by phylogenies inferred from mitochondrially-encoded genes cytochrome c oxidase I and NADH dehydrogenase 4. In summary, our results reveal two cryptic species. Phylogenetic relationships indicate T. dimidiata sensu stricto is not subdivided into monophyletic clades consistent with subspecies. Based on increased support by hypothesis testing, we propose an updated systematic hypothesis for T. dimidiata based on extensive taxon sampling and analysis of both mitochondrial and nuclear genes.
Assuntos
Doença de Chagas/transmissão , Insetos Vetores/classificação , Insetos Vetores/genética , Triatoma/classificação , Triatoma/genética , Animais , América Central , Citocromos b/genética , DNA Espaçador Ribossômico , Genes Mitocondriais , Haplótipos , Humanos , Insetos Vetores/microbiologia , Filogenia , Filogeografia , Triatoma/microbiologia , Trypanosoma cruziRESUMO
Triatoma dimidiata (Latreille, 1811) is the most abundant and significant insect vector of the parasite Trypanosoma cruzi in Central America, and particularly in Guatemala. Tr. cruzi is the causative agent of Chagas disease, and successful disease control requires understanding the geographic distribution and degree of migration of vectors such as T. dimidiata that frequently re-infest houses within months following insecticide application. The population genetic structure of T. dimidiata collected from six villages in southern Guatemala was studied to gain insight into the migration patterns of the insects in this region where populations are largely domestic. This study provided insight into the likelihood of eliminating T. dimidiata by pesticide application as has been observed in some areas for other domestic triatomines such as Triatoma infestans. Genotypes of microsatellite loci for 178 insects from six villages were found to represent five genetic clusters using a Bayesian Markov Chain Monte Carlo method. Individual clusters were found in multiple villages, with multiple clusters in the same house. Although migration occurred, there was statistically significant genetic differentiation among villages (FR T = 0.05) and high genetic differentiation among houses within villages (FSR = 0.11). Relatedness of insects within houses varied from 0 to 0.25, i.e., from unrelated to half-sibs. The results suggest that T. dimidiata in southern Guatemala moves between houses and villages often enough that recolonization is likely, implying the use of insecticides alone is not sufficient for effective control of Chagas disease in this region and more sustainable solutions are required.
Assuntos
Migração Animal , Fluxo Gênico , Insetos Vetores/fisiologia , Repetições de Microssatélites , Triatoma/fisiologia , Animais , Teorema de Bayes , Doença de Chagas/transmissão , Feminino , Guatemala , Humanos , Insetos Vetores/genética , Masculino , Triatoma/genética , Trypanosoma cruzi/fisiologiaRESUMO
BACKGROUND: Triatoma dimidiata, currently the major Central American vector of Trypanosoma cruzi, the parasite that causes Chagas disease, inhabits caves throughout the region. This research investigates the possibility that cave dwelling T. dimidiata might transmit the parasite to humans and links the blood meal sources of cave vectors to cultural practices that differ among locations. METHODOLOGY/PRINCIPAL FINDINGS: We determined the blood meal sources of twenty-four T. dimidiata collected from two locations in Guatemala and one in Belize where human interactions with the caves differ. Blood meal sources were determined by cloning and sequencing PCR products amplified from DNA extracted from the vector abdomen using primers specific for the vertebrate 12S mitochondrial gene. The blood meal sources were inferred by ≥ 99% identity with published sequences. We found 70% of cave-collected T. dimidiata positive for human DNA. The vectors had fed on 10 additional vertebrates with a variety of relationships to humans, including companion animal (dog), food animals (pig, sheep/goat), wild animals (duck, two bat, two opossum species) and commensal animals (mouse, rat). Vectors from all locations fed on humans and commensal animals. The blood meal sources differ among locations, as well as the likelihood of feeding on dog and food animals. Vectors from one location were tested for T. cruzi infection, and 30% (3/10) tested positive, including two positive for human blood meals. CONCLUSIONS/SIGNIFICANCE: Cave dwelling Chagas disease vectors feed on humans and commensal animals as well as dog, food animals and wild animals. Blood meal sources were related to human uses of the caves. We caution that just as T. dimidiata in caves may pose an epidemiological risk, there may be other situations where risk is thought to be minimal, but is not.
Assuntos
Cavernas , Cultura , Insetos Vetores/parasitologia , Natação , Triatoma/fisiologia , Animais , Sequência de Bases , Belize , Doença de Chagas/epidemiologia , Doença de Chagas/parasitologia , Doença de Chagas/transmissão , DNA/genética , Primers do DNA/genética , Vetores de Doenças , Guatemala , Humanos , Reação em Cadeia da Polimerase , Triatoma/parasitologia , Trypanosoma cruzi/genéticaRESUMO
Triatoma dimidiata, a Chagas disease vector distributed in Mexico, Central America, Colombia, Venezuela, Peru and Ecuador, has been studied using genetic markers and four groups have been defined by ITS-2 sequences: 1A, 1B, 2 and 3. To gather evidence on the divergence and reproductive isolation among T. dimidiata ITS-2 groups, we carried out 15 crossbreeding experiments with field-collected sylvan and domestic T. dimidiata from Guatemala where three groups are found: 1A, 2 and 3. Reciprocal crosses between individuals from groups 1A and 2, and a cross between group 2 individuals from different habitats, produced an average 129.78±42.29 eggs with hatching success ranging from 31.6 to 90.1%. The offspring of these crosses reached the adult stage, and crosses between F1 insects produced eggs. These results suggest that there are no pre- or post-zygotic reproductive barriers between groups 1A and 2, or within group 2. Crosses between group 3 females and males from groups 1A or 2 produced on average 85.67±30.26 eggs and none of them hatched. These results support the existence of pre-zygotic barriers between T. dimidiata group 3 and groups 1A and 2. The group 3 individuals were collected in sylvatic environments in Yaxha, Peten, Guatemala. Previously, distinct chromosomal characteristics (cytotype 3) were described in individuals from this population. Based on this evidence we suggest that this population is divergent at the species level from other T. dimidiata populations.
Assuntos
Insetos Vetores , Isolamento Reprodutivo , Triatoma/fisiologia , Animais , América Central , DNA Espaçador Ribossômico/genética , Feminino , Genótipo , Hibridização Genética , Masculino , Triatoma/classificação , Triatoma/genéticaRESUMO
Triatoma dimidiata is the most important Chagas disease insect vector in Central America as this species is primarily responsible for Trypanosoma cruzi transmission to humans, the protozoan parasite that causes Chagas disease. T. dimidiata sensu lato is a genetically diverse assemblage of taxa and effective vector control requires a clear understanding of the geographic distribution and epidemiological importance of its taxa. The nuclear ribosomal internal transcribed spacer 2 (ITS-2) is frequently used to infer the systematics of triatomines. However, oftentimes amplification and sequencing of ITS-2 fails, likely due to both the large polymerase chain reaction (PCR) product and polymerase slippage near the 5' end. To overcome these challenges we have designed new primers that amplify only the 3'-most 200 base pairs of ITS-2. This region distinguishes the ITS-2 group for 100% of known T. dimidiata haplotypes. Furthermore, we have developed a PCR-restriction fragment length polymorphism (RFLP) approach to determine the ITS-2 group, greatly reducing, but not eliminating, the number of amplified products that need to be sequenced. Although there are limitations with this new PCR-RFLP approach, its use will help with understanding the geographic distribution of T. dimidiata taxa and can facilitate other studies characterising the taxa, e.g. their ecology, evolution and epidemiological importance, thus improving vector control.
Assuntos
DNA Espaçador Ribossômico/análise , Insetos Vetores/genética , RNA Ribossômico/análise , Triatoma/genética , Animais , Doença de Chagas/transmissão , Amplificação de Genes/genética , Guatemala , Haplótipos , Insetos Vetores/classificação , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Triatoma/classificaçãoRESUMO
Triatoma dimidiata is the most important Chagas disease insect vector in Central America as this species is primarily responsible for Trypanosoma cruzi transmission to humans, the protozoan parasite that causes Chagas disease. T. dimidiata sensu lato is a genetically diverse assemblage of taxa and effective vector control requires a clear understanding of the geographic distribution and epidemiological importance of its taxa. The nuclear ribosomal internal transcribed spacer 2 (ITS-2) is frequently used to infer the systematics of triatomines. However, oftentimes amplification and sequencing of ITS-2 fails, likely due to both the large polymerase chain reaction (PCR) product and polymerase slippage near the 5' end. To overcome these challenges we have designed new primers that amplify only the 3'-most 200 base pairs of ITS-2. This region distinguishes the ITS-2 group for 100% of known T. dimidiata haplotypes. Furthermore, we have developed a PCR-restriction fragment length polymorphism (RFLP) approach to determine the ITS-2 group, greatly reducing, but not eliminating, the number of amplified products that need to be sequenced. Although there are limitations with this new PCR-RFLP approach, its use will help with understanding the geographic distribution of T. dimidiata taxa and can facilitate other studies characterising the taxa, e.g. their ecology, evolution and epidemiological importance, thus improving vector control.
Assuntos
Animais , DNA Espaçador Ribossômico/análise , Insetos Vetores/genética , RNA Ribossômico/análise , Triatoma/genética , Doença de Chagas/transmissão , Guatemala , Amplificação de Genes/genética , Haplótipos , Insetos Vetores/classificação , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Triatoma/classificaçãoRESUMO
A novel method using vector blood meal sources to assess the impact of control efforts on the risk of transmission of Chagas disease was tested in the village of El Tule, Jutiapa, Guatemala. Control used Ecohealth interventions, where villagers ameliorated the factors identified as most important for transmission. First, after an initial insecticide application, house walls were plastered. Later, bedroom floors were improved and domestic animals were moved outdoors. Only vector blood meal sources revealed the success of the first interventions: human blood meals declined from 38% to 3% after insecticide application and wall plastering. Following all interventions both vector blood meal sources and entomological indices revealed the reduction in transmission risk. These results indicate that vector blood meals may reveal effects of control efforts early on, effects that may not be apparent using traditional entomological indices, and provide further support for the Ecohealth approach to Chagas control in Guatemala.
Assuntos
Sangue/parasitologia , Doença de Chagas/prevenção & controle , Doença de Chagas/transmissão , Insetos Vetores/parasitologia , Avaliação de Programas e Projetos de Saúde/métodos , Triatoma/parasitologia , Animais , Comportamento Alimentar , Guatemala , Habitação , Humanos , Controle de Insetos/métodos , Inseticidas , Programas Nacionais de Saúde , Densidade Demográfica , Fatores de Risco , Fatores de Tempo , Trypanosoma cruzi/patogenicidadeRESUMO
Approximately 10 million people are infected with Trypanosoma cruzi, the causative agent of Chagas disease, which remains the most serious parasitic disease in the Americas. Most people are infected via triatomine vectors. Transmission has been largely halted in South America in areas with predominantly domestic vectors. However, one of the main Chagas vectors in Mesoamerica, Triatoma dimidiata, poses special challenges to control due to its diversity across its large geographic range (from Mexico into northern South America), and peridomestic and sylvatic populations that repopulate houses following pesticide treatment. Recent evidence suggests T. dimidiata may be a complex of species, perhaps including cryptic species; taxonomic ambiguity which confounds control. The nuclear sequence of the internal transcribed spacer 2 (ITS2) of the ribosomal DNA and the mitochondrial cytochrome b (mt cyt b) gene were used to analyze the taxonomy of T. dimidiata from southern Mexico throughout Central America. ITS2 sequence divides T. dimidiata into four taxa. The first three are found mostly localized to specific geographic regions with some overlap: (1) southern Mexico and Guatemala (Group 2); (2) Guatemala, Honduras, El Salvador, Nicaragua, and Costa Rica (Group 1A); (3) and Panama (Group 1B). We extend ITS2 Group 1A south into Costa Rica, Group 2 into southern Guatemala and show the first information on isolates in Belize, identifying Groups 2 and 3 in that country. The fourth group (Group 3), a potential cryptic species, is dispersed across parts of Mexico, Guatemala, and Belize. We show it exists in sympatry with other groups in Peten, Guatemala, and Yucatan, Mexico. Mitochondrial cyt b data supports this putative cryptic species in sympatry with others. However, unlike the clear distinction of the remaining groups by ITS2, the remaining groups are not separated by mt cyt b. This work contributes to an understanding of the taxonomy and population subdivision of T. dimidiata, essential for designing effective control strategies.
Assuntos
Doença de Chagas/epidemiologia , Insetos Vetores/classificação , Triatoma/classificação , Animais , Doença de Chagas/transmissão , Fatores de Confusão Epidemiológicos , DNA Intergênico/genética , DNA Mitocondrial/genética , Ecossistema , Guatemala/epidemiologia , Insetos Vetores/genética , México/epidemiologia , Triatoma/genéticaRESUMO
Seven Triatoma dimidiata (Latreille, 1811) populations from different provinces of Guatemala were compared along with three related triatomine species using the electrophoretic profiles of salivary proteins. The analysis of salivary proteins allowed the separation of two of the species into their respective complexes, phyllosoma (T. pallidipennis) and protracta (T. nitida) (Lent and Wygodzinsky, 1979), whereas T. dimidiata seems slightly separated from either of these. Based on salivary protein profiles, T. dimidiata is most closely related to the cluster including T ryckmani and T. nitida (protracta) and more diverged from T. pallidipennis (phyllosoma). Among Guatemalan T. dimidiata populations, the cave population from Lanquin is separated from the rest of populations analyzed, suggesting that it is in the process of speciation. No difference in protein banding pattern was observed among populations from domestic and peridomestic ecotopes from the same region.