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Made available in DSpace on 2022-04-28T19:46:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-03-01 Agencia Nacional de Investigación e Innovación American University University of Arizona University of Vermont Junta de Andalucía Comisión Sectorial de Investigación Científica Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Ministerio de Educación y Cultura The subfamily Triatominae (Hemiptera-Reduviidae) includes more than 150 blood-sucking species, potential vectors of the protozoan Trypanosoma cruzi, causative agent of Chagas disease. A distinctive cytogenetic characteristic of this group is the presence of extremely stable chromosome numbers. Unexpectedly, the analyses of the chromosomal location of ribosomal gene clusters and other repetitive sequences place Triatominae as a significantly diverse hemipteran subfamily. Here, we advance the understanding of Triatominae chromosomal evolution through the analysis of the 45S rDNA cluster chromosomal location in 92 Triatominae species. We found the 45S rDNA clusters in one to four loci per haploid genome with different chromosomal patterns: On one or two autosomes, on one, two or three sex chromosomes, on the X chromosome plus one to three autosomes. The movement of 45S rDNA clusters is discussed in an evolutionary context. Our results illustrate that rDNA mobility has been relatively common in the past and in recent evolutionary history of the group. The high frequency of rDNA patterns involving autosomes and sex chromosomes among closely related species could affect genetic recombination and the viability of hybrid populations, which suggests that the mobility of rDNA clusters could be a driver of species diversification. Sección Genética Evolutiva Facultad de Ciencias Universidad de la República Department of Experimental Biology Genetics University of Jaén Department of Biomedicine and Biotechnology University of Alcalá Laboratório de Entomologia em Saúde Pública Departamento de Epidemiologia Faculdade de Saúde Pública Universidade de São Paulo Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista “Júlio de Mesquita Filho” (Unesp) Fundação Oswaldo Cruz Laboratorio de Investigación en Genética Evolutiva – LIGE Universidad Pedagógica y Tecnológica de Colombia Programa de Biología Celular y Molecular ICBM Facultad de Medicina Universidad de Chile Laboratorio de Entomología Aplicada y Parasitología Escuela de Biología Facultad de Farmacia Universidad de San Carlos de Guatemala Department of Biological Sciences Loyola University New Orleans Departamento de Microbiología y Parasitología Facultad de Medicina Universidad Nacional Autónoma de México Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista “Júlio de Mesquita Filho” (Unesp) Comisión Sectorial de Investigación Científica: 160 FAPESP: 17/05015-7 FAPESP: 19/02145-2 FAPESP: 307 398/2018-8 Ministerio de Educación y Cultura: II/FVF/2019/054

Despite successes in reducing transmission, Chagas disease (American trypanosomiasis) remains the greatest economic burden of any parasitic disease in Latin America afflicting mostly the poor and further contributing to poverty. We review a long-term (2001-2022), integrated Ecohealth approach that addresses sustainable development goals to reduce risk of Chagas transmission by the main native vector in Central America, Triatoma dimidiata, s.l. The basis of the Ecohealth intervention was the identification of the risk factors for house infestation, an understanding of and collaboration with local communities, and genetic and proteomic studies that revealed the epidemiology and mechanisms of the rapid reinfestation seen following insecticide application. We review the development of this approach from a pilot project in two Guatemalan villages, to an expanded initiative across three countries with vastly different ecology, cultures, and municipal organization, and finally development of a multi-institutional, large-scale project to develop a strategy to tackle the remaining hot spots in Central America. This integrated Ecohealth approach resulted in reduced risk of transmission as measured by a sustained decrease in house infestation without further use of insecticides, a reduction in vectors with human blood meals and the Chagas parasite, as well as other health and economic benefits. We discuss lessons learned and how this approach could be applied to other vector-borne diseases.

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.

Chagas disease is a parasitic infection that can insidiously cause non-ischemic cardiomyopathy. Given the largely silent nature of this progressive disease, asymptomatic blood donors pose potential blood transfusion risk. Blood donation screening has become an unintentional form of Chagas disease surveillance, with thousands of new cases identified since national surveillance was initiated in 2007.We recruited T. cruzi-positive blood donors identified from California and Arizona blood centers for confirmatory blood screening and assessment of lifetime infection risk.Among eight suspected cases, we identified four confirmed US autochthonous infections. The current manuscript details the transmission sources, healthcare-seeking behaviors post-blood donation resulting, and clinical course of disease among persons without any history of travel to endemic Latin American countries.This manuscript presents four additional US-acquired Chagas disease cases and identifies an opportunity for blood centers to assist in confronting barriers surrounding Chagas disease in the US.

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, peri‑domestic 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.

Scientific collections such as the U.S. National Museum (USNM) are critical to filling knowledge gaps in molecular systematics studies. The global taxonomic impediment has resulted in a reduction of expert taxonomists generating new collections of rare or understudied taxa and these large historic collections may be the only reliable source of material for some taxa. Integrated systematics studies using both morphological examinations and DNA sequencing are often required for resolving many taxonomic issues but as DNA methods often require partial or complete destruction of a sample, there are many factors to consider before implementing destructive sampling of specimens within scientific collections. We present a methodology for the use of archive specimens that includes two crucial phases: 1) thoroughly documenting specimens destined for destructive sampling—a process called electronic vouchering, and 2) the pipeline used for whole genome sequencing of archived specimens, from extraction of genomic DNA to assembly of putative genomes with basic annotation. The process is presented for eleven specimens from two different insect subfamilies of medical importance to humans: Anophelinae (Diptera: Culicidae)—mosquitoes and Triatominae (Hemiptera: Reduviidae)—kissing bugs. Assembly of whole mitochondrial genome sequences of all 11 specimens along with the results of an ortholog search and BLAST against the NCBI nucleotide database are also presented.

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.

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.

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.

Kissing bugs in the genus Triatoma are obligate blood feeders that feed mainly on vertebrate blood and have lost the predatory lifestyle found in other reduviid bugs. They occasionally also feed on the hemolymph of arthropods, especially during the first and second instar stages. The largest kissing bug species in the United States, Triatoma recurva (Stal) (Hemiptera: Reduviidae), is poorly known and was chosen to investigate its ability to feed and develop on a diet of cockroach hemolymph. Molting from first instar individuals to second instars readily occurred at approximately the same rate reported for the species feeding on mammalian blood. Subsequent instars also fed on and survived on cockroach hemolymph with some individuals maturing to adults. In the larger instars, development time and survival rates were reduced relative to the results reported in the literature for mammalian-blood-fed individuals. Two other species of kissing bugs, Triatoma protracta (Uhler) and T. rubida (Uhler) failed to survive on cockroach hemolymph with most individuals failing to molt from the first instar stage. Although T. recurva does not thrive on a diet limited to hemolymph of cockroaches, it appears to be an unusual species in which cockroaches might be a primary source of nutrition for smaller individuals and are a viable exclusive source of nutrition for all immatures. At a minimum during times of limited availability of vertebrate blood sources, the presence of cockroaches enhances survival opportunities. Efforts to control populations of this kissing bug species likely will be improved with additional control of cockroach populations in the environment.

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 22 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.

A new species of the genus Triatoma Laporte, 1832 (Hemiptera, Reduviidae) is described based on specimens collected in the department of Huehuetenango, Guatemala. Triatomahuehuetenanguensissp. n. is closely related to T.dimidiata (Latreille, 1811), with the following main morphological differences: lighter color; smaller overall size, including head length; and width and length of the pronotum. Natural Trypanosomacruzi (Chagas, 1909) infection, coupled with its presence in domestic habitats, makes this species a potentially important vector of Trypanosomacruzi in Guatemala.

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., Author summary Chagas disease is caused by the parasite Trypanosoma cruzi, which is spread by triatomine kissing bugs. There are many biotic factors that influence the risk of disease transmission, including the strain of the parasite, the vector movement patterns, the community of microbes interacting with the parasite inside the vector's gut, and the availability of suitable vertebrate hosts. DNA from all of these species can be found in the gut of an infected bug, providing an opportunity to investigate all of them simultaneously by genetically analyzing this single tissue. In this study, we developed a DNA-based method to retrieve, separate, and analyze genetic information from the abdomens of 32 T. dimidiata kissing bug vectors collected across Central America. We found two distinct strains of T. cruzi, and four T. dimidiata genetic clusters associated with environmental and geographical characteristics. These populations harbored different bacterial gut communities that were augmented by specifically infection-associated bacteria when the vector was infected by the parasite. In some cases, we could identify what the insect had recently fed on, including chicken, duck, dog and human. Having simultaneous information on all of these organisms may help to fine-tune control strategies that influence the risk of T. cruzi transmission.

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.

In this paper, Triatoma mopan sp. 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.

Phylogenetic relationships of insect vectors of parasitic diseases are important for understanding the evolution of epidemiologically relevant traits, and may be useful in vector control. The sub-family Triatominae (Hemiptera:Reduviidae) includes ∼140 extant species arranged in five tribes comprised of 15 genera. The genus Triatoma is the most species-rich and contains important vectors of Trypanosoma cruzi, the causative agent of Chagas disease. Triatoma species were grouped into complexes originally by morphology and more recently with the addition of information from molecular phylogenetics (the four-complex hypothesis); however, without a strict adherence to monophyly. To date, the validity of proposed species complexes has not been tested by statistical tests of topology. The goal of this study was to clarify the systematics of 19 Triatoma species from North and Central America. We inferred their evolutionary relatedness using two independent data sets: the complete nuclear internal transcribed spacer-2 ribosomal DNA (ITS-2 rDNA) and head morphometrics. In addition, we used the Shimodaira-Hasegawa statistical test of topology to assess the fit of the data to a set of competing systematic hypotheses (topologies). An unconstrained topology inferred from the ITS-2 data was compared to topologies constrained based on the four-complex hypothesis or one inferred from our morphometry results. The unconstrained topology represents a statistically significant better fit of the molecular data than either the four-complex or the morphometric topology. We propose an update to the composition of species complexes in the North and Central American Triatoma, based on a phylogeny inferred from ITS-2 as a first step towards updating the phylogeny of the complexes based on monophyly and statistical tests of topologies.

Background Kissing bugs, vectors of Trypanosoma cruzi , the parasite that causes Chagas disease, are common in the desert Southwest. After a dispersal flight in summer, adult kissing bugs occasionally gain access to houses where they remain feeding on humans and pets. How often wild, free-roaming kissing bugs feed on humans outside their homes has not been studied. This is important because contact of kissing bugs with humans is one means of gauging the risk for acquisition of Chagas disease. Methods We captured kissing bugs in a zoological park near Tucson, Arizona, where many potential vertebrate hosts are on display, as well as being visited by more than 300,000 humans annually. Cloacal contents of the bugs were investigated for sources of blood meals and infection with T. cruzi . Results Eight of 134 captured bugs were randomly selected and investigated. All 8 (100%) had human blood in their cloacae, and 7 of 8 (88%) had fed on various vertebrates on display or feral in the park. Three bugs (38%) were infected with T. cruzi . Three specimens of the largest species of kissing bug in the United States ( Triatoma recurva ) were captured in a cave and walking on a road; 2 of 3 (67%) had fed on humans. No T. recurva harbored T. cruzi . Conclusions This study establishes that free-roaming kissing bugs, given the opportunity, frequently feed on humans outside the confines of their homes in the desert Southwest and that some harbored T. cruzi . This could represent a hitherto unrecognized potential for transmission of Chagas disease in the United States.

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.

A high proportion of triatomine insects, vectors for Trypanosoma cruzi trypanosomes, collected in Arizona and California and examined using a novel assay had fed on humans. Other triatomine insects were positive for T. cruzi parasite infection, which indicates that the potential exists for vector transmission of Chagas disease in the United States.

From May through November 2007, intensive weekly surveys at the site of a previously reported autochthonous human case of Chagas parasite infection resulted in the collection of 298 Triatoma sanguisuga (Leconte) specimens, of which 60.4% (180) were polymerase chain reaction positive for Trypanosoma cruzi Chagas. All were adults, in a ratio of approximately 1:1 female to male, indicating that the domicile was not colonized, but was a destination for these host-seeking adults. We report on seasonal activity pattern, T. cruzi prevalence in T. sanguisuga, and attempts at insect exclusion and control at the case residence.

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.

Evidence of the compressive growth of basilar-membrane displacement can be seen in distortion-product otoacoustic emission (DPOAE) levels measured as a function of stimulus level. When the levels of the two stimulus tones (f1 and f2) are related by the formula L1 = 39 dB + 0.4 x L2 [Kummer et al., J. Acoust. Soc. Am. 103, 3431-3444 (1998)] the shape of the function relating DPOAE level to L2 is similar (up to an L2 of 70 dB SPL) to the classic Fletcher and Munson [J. Acoust. Soc. Am. 9, 1-10 (1933)] loudness function when plotted on a logarithmic scale. Explicit estimates of compression have been derived based on recent DPOAE measurements from the laboratory. If DPOAE growth rate is defined as the slope of the DPOAE I/O function (in dB/dB), then a cogent definition of compression is the reciprocal of the growth rate. In humans with normal hearing, compression varies from about 1 at threshold to about 4 at 70 dB SPL. With hearing loss, compression is still about 1 at threshold, but grows more slowly above threshold. Median DPOAE I/O data from ears with normal hearing, mild loss, and moderate loss are each well fit by log functions. When the I/O function is logarithmic, then the corresponding compression is a linear function of stimulus level. Evidence of cochlear compression also exists in DPOAE suppression tuning curves, which indicate the level of a third stimulus tone (f3) that reduces DPOAE level by 3 dB. All three stimulus tones generate compressive growth within the cochlea; however, only the relative compression (RC) of the primary and suppressor responses is observable in DPOAE suppression data. An RC value of 1 indicates that the cochlear responses to the primary and suppressor components grow at the same rate. In normal ears, RC rises to 4, when f3 is an octave below f2. The similarities between DPOAE and loudness compression estimates suggest the possibility of predicting loudness growth from DPOAEs; however, intersubject variability makes such predictions difficult at this time.

Trypanosoma cruzi, the hemoflagellate parasite and cause of Chagas disease in Latin America, is carried by Triatomine vectors, principally Triatoma dimidiata and Rhodnius prolixus in Central America. To assist control efforts and to understand the epidemiology of the disease in Guatemala, the population genetics of T. dimidiata was analyzed among three houses within a village and two adjacent villages in Guatemala. Eleven Randomly Amplified Polymorphic DNA-polymerase chain reaction (RAPD-PCR) primers were screened and three used to amplify bands, 29 of which were scored, from T. dimidiata DNA of ≈50 bugs per house from three houses within a village and from 66 and 33 bugs, respectively, from adjacent villages. Results show very small genetic distances among the three T. dimidiata subpopulations from the houses (D = 0.013–0.022) and the two villages (D = 0.0199). The amount of differentiation among houses (fixation index, FST) was also very small, FST = 0.025 among the houses and the two villages ...

Measurements of DPOAE level in the presence of a suppressor were used to describe a pattern that is qualitatively similar to population studies in the auditory nerve and to behavioral studies of upward spread of masking. DPOAEs were measured in the presence of a suppressor (f3) fixed at either 2.1 or 4.2 kHz, and set to each of seven levels (L3) from 20 to 80 dB SPL. In the presence of a fixed f3 and L3 combination, f2 was varied from about 1 oct below to at least 1/2 oct above f3, while L2 was set to each of 6 values (20-70 dB SPL). L1 was set according to the equation L1 = 0.4L2 + 39 [Janssen et al., J. Acoust. Soc. Am. 103, 3418-3430 (1998)]. At each L2, L1 combination, DPOAE level was measured in a control condition in which no suppressor was presented. Data were converted into decrements (the amount of suppression, in dB) by subtracting the DPOAE level in the presence of each suppressor from the DPOAE level in the corresponding control condition. Plots of DPOAE decrements as a function of f2 showed maximum suppression when f2 approximately = f3. As L3 increased, the suppressive effect spread more towards higher f2 frequencies, with less spread towards lower frequencies relative to f3. DPOAE decrement versus L3 functions had steeper slopes when f2f3, compared to the slopes when f2f3. These data are consistent with other findings that have shown that response growth for a characteristic place (CP) or frequency (CF) depends on the relation between CP or CF and driver frequency, with steeper slopes when driver frequency is less than CF and shallower slopes when driver frequency is greater than CF. For a fixed amount of suppression (3 dB), L3 and L2 varied nearly linearly for conditions in which f3 approximately = f2, but grew more rapidly for conditions in which f3f2, reflecting the basal spread of excitation to the suppressor. The present data are similar in form to the results observed in population studies from the auditory nerve of lower animals and in behavioral masking studies in humans.

Although rare, there have been isolated reports of autochthonous transmission of Trypanosoma cruzi Chagas in the United States. In June 2006, a human case of domestically transmitted T. cruzi was identified in southern Louisiana. To examine the localized risk of human T. cruzi infection in the area surrounding the initial human case, environmental surveys of households in the area and a serological survey of the residents were performed between September 2008 and November 2009. Human T. cruzi infection was determined using a rapid antigen field test, followed by confirmatory enzyme-linked immunosorbent assay testing in the laboratory. A perimeter search of each participating residence for Triatoma sanguisuga (LeConte), the predominant local triatomine species, was also performed. No participating individuals were positive for antibodies against T. cruzi; however, high levels of T. cruzi infection (62.4%) were detected in collected T. sanguisuga. Households with T. sanguisuga presence were less likely to use air conditioning, and more likely to have either chickens or cats on the property. While the human risk for T cruzi infection in southeastern Louisiana is low, a high prevalence of infected T. sanguisuga does indicate a substantial latent risk for T. cruzi peridomestic transmission. Further examination of the behavior and ecology of T. sanguisuga in the region will assist in refining local T. cruzi risk associations.

Primary and secondary sources combine to produce the 2f1-f2 distortion product otoacoustic emission (DPOAE) measured in the ear canals of humans. DPOAEs were obtained in nine normal-hearing subjects using a fixed-f2 paradigm in which f1 was varied. The f2 was 2 or 4 kHz, and absolute and relative primary levels were varied. Data were obtained with and without a third tone (f3) placed 15.6 Hz below 2f1-f2. The level of f3 was varied in order to suppress the stimulus frequency otoacoustic emission (SFOAE) coming from the 2f1-f2 place. These data were converted from the complex frequency domain into an equivalent time representation using an inverse fast Fourier transform (IFFT). IFFTs of unsuppressed DPOAE data were characterized by two or more peaks. Relative amplitudes of these peaks depended on overall primary level and on primary-level differences. The suppressor eliminated later peaks, but early peaks remained relatively unaltered. Results are interpreted to mean that the DPOAE measured in humans includes components from the f2 place (intermodulation distortion) and DP place (in the form of a SFOAE). These findings build on previous work by providing evidence that multiple peaks in the IFFT are due to a secondary source at the DP place.

Through the Southern Cone Initia-tive, Chagas disease was effectively ‘whac’d’ byintergovernmental partnerships supervised by thePan American Health Organization (PAHO),and transmission halted in Uruguay (1997),Chile (1999), and in most areas of Argentina,Paraguay and Bolivia, by eliminating the insectvectors and blood transfusion transmission.Called the silent killer, Chagas disease usuallydemonstrates mild symptoms during the initialweeks following infection (swelling at the site ofinfection, fever, malaise and lymphadenopathy),so diagnosis at this stage is rare. Most patientsprogress to the ‘silent’, sometimes decades-longasymptomatic period before 20–40% of infectedpeople advance to fatal heart disease and/ormegasyndrome. Around 80% of transmissionoccurs via insect vectors; parasites in the fecestypically gain access through the bite wound ormucous membranes. Transmission by bloodproducts and solid-organ transplantation, con-genitally and even by contaminated food or drinkalso occurs. Different strains of

Interactions among age, threshold, and frequency in relation to distortion product otoacoustic emissions (DPOAE) have yet to be resolved. The effects of these variables were explored by analyzing DPOAEs in ears with thresholds not exceeding 20 dB HL. Multivariate regression analyses were performed in two different ways. For data to be included in the first analysis, audiometric threshold had to be 20 dB HL or better only at the particular frequency under study, but might exceed 20 dB HL at other half-octave frequencies. Significant main effects were found for age, threshold, and frequency. There was also an age-by-frequency interaction, but a significant age-by-threshold interaction was not observed. DPOAE amplitudes decreased as either age, frequency, or threshold increased. In the second analysis, when a more stringent inclusion criterion was applied (normal thresholds at all frequencies), the main effects for age, threshold, and frequency were not significant. The significant age-by-frequency interaction remained, whereby DPOAE amplitudes decreased as age and frequency increased, but the age-by-threshold interaction again was not significant. The magnitude of DPOAE amplitude change across age, threshold, and frequency and for the age-by-frequency interaction was small but similar for both groups of subjects. Age in association with threshold did not account for observed changes in DPOAE amplitudes for either group. Importantly, the lack of a significant age-by-threshold interaction indicates that there may be processes intrinsic to aging alone that act on DPOAE generation.

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.

International audience; Triatomines are hemipteran bugs acting as vectors of the protozoan parasite Trypanosoma cruzi. This parasite causes Chagas disease, one of the major parasitic diseases in the Americas. Studies of triatomine genetics and evolution have been particularly useful in the design of rational vector control strategies, and are reviewed here. The phylogeography of several triatomine species is now slowly emerging, and the struggle to reconcile the phenotypic, phylogenetic, ecological and epidemiological species concepts makes for a very dynamic field. Population genetic studies using different markers indicate a wide range of population structures, depending on the triatomine species, ranging from highly fragmented to mobile, interbreeding populations. Triatomines transmit T. cruzi in the context of complex interactions between the insect vectors, their bacterial symbionts and the parasites; however, an integrated view of the significance of these interactions in triatomine biology, evolution and in disease transmission is still lacking. The development of novel genetic markers, together with the ongoing sequencing of the Rhodnius prolixus genome and more integrative studies, will provide key tools to expanding our understanding of these important insect vectors and allow the design of improved vector control strategies.Heredity advance online publication, 7 September 2011; doi:10.1038/hdy.2011.71.

PCR detection of Trypanosoma cruzi in Rhodnius prolixus using fresh tissue or fecal drops on filter paper showed comparable results: 38.7% infection rate using the fresh tissue sample and 37.9% by dried fecal drop.

Chagas disease remains endemic across much of Latin America, but is largely enzootic--restricted to wild mammals and triatomine vectors in the United States. Within the United States, there are ten recognized species of triatomines and 18 mammals reported naturally infected with Trypanosoma cruzi, the causative agent of Chagas disease. However, only six cases of autochthonous vector-borne transmission of T. cruzi to humans have been reported in the United States. As a follow-up to the sixth reported case, triatomine insects were collected from the index case site, in a rural area of New Orleans, LA, USA. During the summer months of 2006 and 2007, 344 Triatoma sanguisuga were collected and showed a T. cruzi infection prevalence of 56%. A subset of these insects was analyzed to infer intraspecific genetic variation from a 606 bp fragment of cytochrome b (n=54) and a 340 bp fragment of 16S ribosomal DNA (n=17). From the 54 samples, 37 cytb haplotypes (H(d)=0.978) were observed and 14.7% of positions were polymorphic. Phylogenetic analysis divides the population into two distinct groups with an average pairwise genetic distance of ~5%. The 16S rDNA sequences revealed 6 haplotypes among 17 specimens (H(d)=0.713) with 1.2% of the positions exhibiting polymorphisms. 16S polymorphism data support the concept of two groups within this single population. The genetic distance of Group 1 from Group 2 suggests that Group 1 could represent a sub-species as this level of divergence is similar to that observed among other triatomine subspecies.