Chagas Disease In Venezuela Research Articles

Parasitological and molecular characterization of Trypanosoma cruzi of triatomines and mammals from endemic areas for Chagas disease in Venezuela.

It is estimated that 6-7 million people worldwide are infected with Trypanosoma cruzi, the parasite that causes Chagas disease. In Venezuela, Chagas disease remains a public health problem. In this work, T. cruzi isolates from six species of triatomines and mammals of the orders Didelphimorphia and Xenarthra, captured in rural communities of Monagas, underwent parasitological and molecular characterization. A total of 471 triatomines and 17 mammals were captured, with a natural infection rate of 41.4% and 70.6%, respectively. In the male NMRI mouse model used for parasitological characterization (prepatent period, parasitemia curve, mouse mortality, and tissular parasitism), T. cruzi isolates exhibited high lethality due to their pronounced virulence, irrespective of the parasite load in each mouse, resulting in a mortality rate of 75%. Among the vector isolates, in the mouse model, only 2 out of 6 remained alive, while the rest perished during the evaluation. Conversely, the isolates from mammals proved fatal for all the inoculated mice. All isolates were identified as belonging to DTU TcI, based on the molecular markers such as the intergenic region of the miniexon, D7 divergent domain of the 24Sα rDNA, size-variable domain of the 18S rDNA, and hsp60-PCR-RFLP-EcoRV. This study demonstrates the presence of vectors and mammalian reservoirs naturally infected with T. cruzi in communities of Monagas, the 9th largest and 9th most populous state in Venezuela. This situation represents a neglected epidemiological problem demanding urgent attention and imperative health care intervention.

Tele-entomology and tele-parasitology: A citizen science-based approach for surveillance and control of Chagas disease in Venezuela

Chagas Disease (CD), a chronic infection caused by the Trypanosoma cruzi parasite, is a Neglected Tropical Disease endemic to Latin America. With a re-emergence in Venezuela during the past two decades, the spread of CD has proved susceptible to, and inhibitable by a digital, real-time surveillance system effectuated by Citizen Scientists in communities throughout the country. The #TraeTuChipo (#BringYourKissingBug) campaign implemented in January 2020, has served as such a strategy counting on community engagement to define the current ecological distribution of CD vectors despite the absence of a functional national surveillance program. This pilot campaign collected data through online surveys, social media platforms, and/or telephone text messages. A total of 79 triatomine bugs were reported from eighteen Venezuelan states; 67 bugs were identified as Panstrongylus geniculatus, 1 as Rhodnius pictipes, 1 as Triatoma dimidiata, and 10 as Triatoma maculata. We analyzed 8 triatomine feces samples spotted from 4 Panstrongylus geniculatus which were confirmed positive by qPCR for T. cruzi. Further molecular characterization of discrete typing units (DTUs), revealed that all samples contained TcI, the most highly diverse and broadly distributed strain of T. cruzi. Moreover, analysis of the mitochondrial 12S gene revealed Myotis keaysi, Homo sapiens, and Gallus gallus as the main triatomine feeding sources. This study highlights a novel Citizen Science approach which may help improve the surveillance systems for CD in endemic countries.

Polymorphisms of the TLR4 gene: Risk factor for chronicity and severity in oral vectorial Chagas disease

Chagas disease is one of the parasitic infections with the greatest socio-economic impact in Latin America. In Venezuela, epidemiological data has shown different sources of infection, such as the vectorial route by oral transmission. Given the importance of the TLR4 gene in the innate immune response triggered by infection with Trypanosoma cruzi, this work analyses the role of TLR4 polymorphisms and its possible effect on cytokine expression. Genomic DNA was extracted from the peripheral blood of patients from the main outbreak of oral Chagas disease in Venezuela (n = 90), as well as from a group of healthy individuals (n = 183). Subsequently, peripheral blood was also extracted from individuals with different TLR4 haplotypes and then stimulated with LPS to determine the cytokine concentration by ELISA. The internalization of TLR4 was evaluated by flow cytometry. In comparison to healthy individuals, the analysis showed a significantly increased frequency of the Asp/Gly genotype in symptomatic patients. Also, observed a correlation of the 299/399 haplotype with a significant decrease in cytokine concentration and disease severity. Finally, the parasites’ trypomastigotes cause the internalization or negative regulation of TLR4. The variants of TLR4 associated with low production of cytokines may be a risk factor for chronicity and severity (cardiac involvement) in oral vectorial Chagas disease.

Predictions of Future Geographical Distribution of Two Vectors of American Trypanosomiasis: Implications for Endemic Chagas disease in Texas, USA

It is known that climate has a direct effect on vectorborne and zoonotic diseases, and in the face of climate change, understanding this link has become more urgent. Many such vector-borne diseases primarily afflict impoverished populations and have therefore been previously understudied. One major focus of our research is to understand the influence that climate has on the distribution of disease causing microorganisms and their vectors, especially those in relation to American trypanosomiasis (Chagas disease). Chagas disease is caused by the hemoflagellate protozoan parasite, Trypanosoma cruzi. For this study, we hypothesized that the increasing prevalence Chagas in the state of Texas is due to expanding distributions of vectors. To test this hypothesis, historical data on vector distribution and climate was used to determine the probable locations of prevalent vectors in Texas. Predictions for the future distributions were made using environmental niche models for bioclimatic variables with a maximum entropy algorithm. Of the two Triatominae species studied, the range and concentration of both decreased under a global warming scenario, a finding that is consistent with the current research of risk of Chagas disease in Venezuela. In future, this same procedure will be used on more Chagas vectors to better understand if there is a northward shift for vectors, or if Texas is becoming more inhospitable to all vectors of Chagas.

Rhodnius prolixus intoxicated

Rhodnius prolixus (Hemiptera: Reduviidae) is a hematophagous insect native from South America. By the end of the 20th century, it was one of the main vectors of Chagas disease in Venezuela, Colombia, several Central American countries and southern Mexico. The aim of the present article is to review the literature regarding R. prolixus toxicology. British entomologist Vincent B. Wigglesworth carried out the first studies on this subject over seventy years ago. A wide bibliographical search allowed to locate one hundred and thirty scientific articles describing the effects of different insecticides on R. prolixus. About one-third of these articles report the acute toxicity and/or sublethal effects produced by the main synthetic neurotoxic families of insecticides (organochlorines, organophosphates, carbamates and pyrethroids). Only a couple of these studies have regarded the toxicokinetics or toxicodynamics of these insecticides. Insect growth or development disruptors, such as juvenoids, chitin synthesis inhibitors, precocenes, azadirachtin and lignoids, have been thoroughly studied in R. prolixus. Important aspects on the mode of action of ureases were also described in this species. By the end of the 1960’s, resistance to insecticides was detected in R. prolixus from Venezuela. Some years later, the existence of pyrethroid-resistant individuals was also reported. Control programmes for R. prolixus in countries where Chagas is endemic have only used synthetic neurotoxic insecticides. In 2011, Central America and southern Mexico were declared free of this insect. The recent sequencing of the R. prolixus genome will provide valuable information to understand the molecular basis of insecticide resistance in this species.

Evaluación de la Patogenicidad Inhalatoria del Micoinsecticida Beauveria bassiana LF14 en Ratones

<p>Se realizaron pruebas de bioseguridad para evaluar los posibles efectos patogénicos del aislamiento autóctono LF14 de <em>Beauveria bassiana</em> en ratones albinos NMRI; el cual posee una elevada virulencia hacia <em>Rhodnius prolixus</em> y <em>Triatoma maculata</em> (Triatominae), vectores de enfermedad de Chagas en Venezuela. Se administró 1x10<sup>9</sup> conidias/ml del hongo mediante gotas nasales. Se realizaron observaciones clínicas y de comportamiento, así como de peso corporal en forma diaria y por 14 días. Asimismo, se estimó el aclaramiento fúngico mediante examen directo y cultivo de muestras intranasales, y de infectividad con la toma de muestras de tejidos de varios órganos, incluyendo un estudio histopatológico a los 3, 7 y 14 días después de la inoculación. No ocurrieron muertes ni alteraciones clínico-patológicas discernibles, siendo el tipo de comportamiento activo-normal en el 100% de los casos. El peso promedio corporal se incrementó, tanto en los animales expuestos a LF14 <em>B. bassiana</em> como los controles, habiendo diferencias entre sexos (p<0.0000). No se detectaron anormalidades en la inspección anatomopatológica ni hubo reacciones tisulares de consideración. Solo se obtuvieron conidias viables sin germinar al día tres de la necropsia en pulmón y tráquea, sugiriendo que no existe evidencia de multiplicación fúngica. Los resultados obtenidos permiten sugerir que el aislamiento <em>B. bassiana</em> LF14 no es patogénico por vía inhalatoria en <em>Mus musculus</em> (NMRI).</p>

The impact of climate change on the geographical distribution of two vectors of Chagas disease: implications for the force of infection.

Chagas disease, caused by the parasite Trypanosoma cruzi, is the most important vector-borne disease in Latin America. The vectors are insects belonging to the Triatominae (Hemiptera, Reduviidae), and are widely distributed in the Americas. Here, we assess the implications of climatic projections for 2050 on the geographical footprint of two of the main Chagas disease vectors: Rhodnius prolixus (tropical species) and Triatoma infestans (temperate species). We estimated the epidemiological implications of current to future transitions in the climatic niche in terms of changes in the force of infection (FOI) on the rural population of two countries: Venezuela (tropical) and Argentina (temperate). The climatic projections for 2050 showed heterogeneous impact on the climatic niches of both vector species, with a decreasing trend of suitability of areas that are currently at high-to-moderate transmission risk. Consequently, climatic projections affected differently the FOI for Chagas disease in Venezuela and Argentina. Despite the heterogeneous results, our main conclusions point out a decreasing trend in the number of new cases of Tr. cruzi human infections per year between current and future conditions using a climatic niche approach.

Molecular Epidemiologic Source Tracking of Orally Transmitted Chagas Disease, Venezuela

Oral outbreaks of Chagas disease are increasingly reported in Latin America. The transitory presence of Trypanosoma cruzi parasites within contaminated foods, and the rapid consumption of those foods, precludes precise identification of outbreak origin. We report source attribution for 2 peri-urban oral outbreaks of Chagas disease in Venezuela via high resolution microsatellite typing.

Trypanosoma cruzi genotyping supports a common source of infection in a school-related oral outbreak of acute Chagas disease in Venezuela.

Trypanosoma cruzi I, a discrete typing unit (DTU) found in human infections in Venezuela and other countries of the northern region of South America and in Central America, has been recently classified into five intra-DTU genotypes (Ia, Ib, Ic, Id, Ie) based on sequence polymorphisms found in the spliced leader intergenic region. In this paper we report the genotype identification of T. cruzi human isolates from one outbreak of acute orally acquired Chagas disease that occurred in a non-endemic region of Venezuela and from T. cruzi triatomine and rat isolates captured at a guava juice preparation site which was identified as the presumptive source of infection. The genotyping of all these isolates as TcId supports the view of a common source of infection in this oral Chagas disease outbreak through the ingestion of guava juice. Implications for clinical manifestations and dynamics of transmission cycles are discussed.

A panel of ten microsatellite loci for the Chagas disease vector Rhodnius prolixus (Hemiptera: Reduviidae)

Rhodnius prolixus is the main vector of Chagas disease in Venezuela, where it is found colonising rural housing consisting of unplastered adobe walls with palm and/or metal roofs. Vector control failure in Venezuela may be due to the invasion of houses by silvatic populations of R. prolixus found in palms. As part of a study to determine if domestic and silvatic populations of R. prolixus are isolated, thus clarifying the role of silvatic populations in maintaining house infestations, we constructed three partial genomic microsatellite libraries. A panel of ten dinucleotide polymorphic microsatellite markers was selected for genotyping. Allele numbers per locus ranged from three to twelve, with observed and expected heterozygosity ranging from 0.26 to 0.55 and 0.32 to 0.66. The microsatellite markers presented here will contribute to the control of Chagas disease in Venezuela and Colombia through the provision of population information that may allow the design of improved control strategies.

Molecular Genetics Reveal That Silvatic Rhodnius prolixus Do Colonise Rural Houses

Background Rhodnius prolixus is the main vector of Chagas disease in Venezuela. Here, domestic infestations of poor quality rural housing have persisted despite four decades of vector control. This is in contrast to the Southern Cone region of South America, where the main vector, Triatoma infestans, has been eliminated over large areas. The repeated colonisation of houses by silvatic populations of R. prolixus potentially explains the control difficulties. However, controversy surrounds the existence of silvatic R. prolixus: it has been suggested that all silvatic populations are in fact Rhodnius robustus, a related species of minor epidemiological importance. Here we investigate, by direct sequencing (mtcytb, D2) and by microsatellite analysis, 1) the identity of silvatic Rhodnius and 2) whether silvatic populations of Rhodnius are isolated from domestic populations.Methods and FindingsDirect sequencing confirmed the presence of R. prolixus in palms and that silvatic bugs can colonise houses, with house and palm specimens sharing seven cytb haplotypes. Additionally, mitochondrial introgression was detected between R. robustus and R. prolixus, indicating a previous hybridisation event. The use of ten polymorphic microsatellite loci revealed a lack of genetic structure between silvatic and domestic ecotopes (non-significant FST values), which is indicative of unrestricted gene flow.ConclusionsOur analyses demonstrate that silvatic R. prolixus presents an unquestionable threat to the control of Chagas disease in Venezuela. The design of improved control strategies is essential for successful long term control and could include modified spraying and surveillance practices, together with housing improvements.

Morphometric evidence for a possible role of Rhodnius prolixus from palm trees in house re-infestation in the State of Barinas (Venezuela)

The main vector of Chagas disease in Venezuela is Rhodnius prolixus. Specimens of Rhodnius sp., identified elsewhere as R. prolixus by molecular tools, were collected in Barinas State (Venezuela) before insecticide application, and compared by morphometric techniques with post-spraying, re-infesting insects after control. Geometric morphometry was applied, allowing separate analyses of shape and size. The idea of a single species sharing silvatic and domestic/peri-domestic ecotopes was supported, suggesting new evolutionary scenarios for the controversial R. prolixus origins. The same data allowed to understand the possible mechanisms of villages re-infestation after a control campaign, either recolonization by local recovery of survivors or re-infestation by external migrants. Both mechanisms were apparent. Although shape properties could distinguish very close subpopulations such as insects from houses and insects from peri-domestic shelters, they were unable to identify the insects from palm trees as a distinct subpopulation. This strongly suggested that human environment could receive immigrants from palm trees. The pattern of size variation supported the hypothesis of a one-way exchange from silvatic to human environments, but did not support the reverse movement. Thus, morphometric data indicated that a silvatic population of the local vector is probably responsible for re-infesting villages after insecticide application, and they also pointed to the existence of re-infestation by local recovery of survivors. According to this interpretation, new epidemiological scenarios must be considered to improve Chagas disease control in Venezuela.

Exposure to Citral, Cinnamon and Ruda Disrupts the Life Cycle of a Vector of Chagas Disease

The main vector of Chagas disease in Venezuela was exposed to the odors of citral, cinnamon and ruda. Cinnamon was found to stop the life cycle of Rhodnius prolixus relative to untreated animals. Citral and ruda also influenced the life cycle but not to the extent of animals exposed to cinnamon. We suggest that future research be directed toward using cinnamon in field and toxicity tests.

Psychology of Learning: A New Approach to Study Behavior of Rhodnius Prolixus Stal under Laboratory Conditions

Conditioning methodologies associated with the psychology of learning are suggested as a new strategy to investigate behavior of the assassin bug Rhodnius prolixus, which is the main vector of Chagas disease in Venezuela. Chagas disease is the fourth leading cause of death in Latin America, as it causes severe chronic illness and approximately 43,000 deaths per year. To illustrate this strategy, two preliminary experiments are reported. In the first, Pavlovian conditioning was examined by pairing an olfactory conditioned stimulus with a temperature unconditioned stimulus. A temperature of 42 degrees C elicits a complex behavioral sequence in R. prolixus consisting of proboscis extension and crawling. Over the course of 12 training trials, this behavioral sequence was not elicited by an olfactory conditioned stimulus. In the second experiment, a latent inhibition paradigm was used to pre-expose R. prolixus to an olfactory conditioned stimulus before pairing the odor with temperature. Over the course of training, an effect of pre-exposure was found. Suggestions for research are discussed and potential conditioned and unconditioned stimuli identified.

Update on Chagas disease in Venezuela: a review

The present article reviews the status of Chagas disease in Venezuela based on the detection of Trypanosoma cruzi infections both in referred patients with clinical presumptive diagnosis (1988-2002) and in individuals sampled from rural localities representative of the different geographical regions of the country (1995-2002). In the former group from 306 individuals examined, 174 (56.8%) were seropositive to T. cruzi; 73 (42%) in the acute phase with 52 (71%) showing blood circulating parasites, and from these 38% were children under 10 years old. The other 101 (58%) showed chronic infection at different degrees of cardiac complication. In addition, serologic examination of 3835 individuals from rural areas revealed 11.7% seroprevalence. From these, 8.5% (38/448) were children aged from 0 to 10 years old. These figures suggest that Chagas disease may be re-emerging in Venezuela judging for the active transmission detected during the last decade. The success of the Venezuelan anti-chagasic campaign during the last 40 years is evaluated in the frame of the present results. The epidemiological situation is discussed and recommendation to consider Chagas disease as a national priority is given.

Interrupting Chagas disease transmission in Venezuela

The interruption of vectorial transmission of Chagas disease in Venezuela is attributed to the combined effects of ongoing entomoepidemiological surveillance, ongoing house spraying with residual insecticides and the concurrent building and modification of rural houses in endemic areas during almost five decades. The original endemic areas which totaled 750,000 km(2), have been reduced to 365,000 km(2). During 1958-1968, initial entomological evaluations carried out showed that the house infestation index ranged between 60-80%, the house infection index at 8-11% and a house density index of 30-50 triatomine bugs per house. By 1990-98, these indexes were further reduced to 1.6-4.0%, 0.01-0.6% and 3-4 bugs per house respectively. The overall rural population seroprevalence has declined from 44.5% (95% C.I.: 43.4-45.3%) to 9.2% (95% C.I.: 9.0-9.4%) for successive grouped periods from 1958 to 1998. The annual blood donor prevalence is firmly established below 1%. The population at risk of infection has been estimated to be less than four million. Given that prevalence rates are stable and appropriate for public health programmes, consideration has been given to potential biases that may distort results such as: a) geographical differences in illness or longevity of patients; b) variations in levels of ascertainment; c) variations in diagnostic criteria; and d) variations in population structure, mainly due to appreciable population migration. The endemic areas with continuous transmission are now mainly confined to piedmonts, as well as patchy foci in higher mountainous ranges, where the exclusive vector is Rhodnius prolixus. There is also an unstable area, of which landscapes are made up of grasslands with scattered broad-leaved evergreen trees and costal plains, where transmission is very low and occasional outbreaks are reported.

Isozyme variability and differentiation between Rhodnius prolixus, R.robustus and R.pictipes, vectors of Chagas disease in Venezuela.

Enzyme polymorphism in triatomine bugs of the genus Rhodnius (Hemiptera: Reduviidae), vectors of Chagas disease, is analysed using both starch and polyacrylamide gel electrophoresis. Out of forty-five enzymes assayed, the electromorphs of seventeen of them: AO, CA, DIA, ES, ES-A, FH, GPD, G6PD, GPI, MDH, ME, 6PGD, PGM, ACON, ACPH, LAP and SOD, involving twenty-two putative structural loci, were scorable. These gene-enzyme systems were therefore selected for routine characterization of R.prolixus Stal adults from different strains. The first thirteen enzymes, involving sixteen structural loci, were also analysed in first instar nymphs of the three species, R.prolixus, R.robustus Larrousse and R.pictipes Stal. Allelic frequencies were calculated for three R.prolixus strains: three to five loci appeared to be polymorphic. The proportion of polymorphic loci (22%) and the average heterozygosity (0.06) indicated low genetic variability, with significant differences between the strains at individual loci. Rhodnius prolixus and R.robustus were found to have identical isozymic patterns. R.pictipes was genetically well differentiated, with twelve diagnostic loci.

Rural housing for control of Chagas disease in Venezuela.

The home is an important protective element for the health of its inhabitants - but these inhabitants often include not only the householders but also domestic pests and vectors of disease. This is particularly so in Latin America where domestic triatomine bugs thrive in many of the poorer quality rural houses, emerging from their crevices at night to feed and transmit Trypanosoma cruzi in their faeces. At the public health level, there is neither drug nor vaccine suitable for controlling T. cruzi - causative agent of Chagas disease - but transmission can be interrupted by control of the domestic vectors. Traditionally, vector control has involved spraying houses with residual insecticides, but a more long-term solution, with many colateral benefits, is to improve rural housing in such a way that colonization by triatomine bugs is inhibited. Such an approach involves development of low-cost techniques for house construction, and mobilization of rural communities to make use of them. In this, Venezuela has played a leading role, as Roberto Briceno-Leon reports.

Long-term control of Chagas disease in Venezuela: effects on serologic findings, electrocardiographic abnormalities, and clinical outcome.

To evaluate the long-term effects (20 years) of a Chagas control program (CCP) in Venezuela, a prospective serologic evaluation was carried out from 1981 to 1984 on 5771 inhabitants (8%) of Roscio county. This region was selected as a representative area where the national CCP was implemented effectively. Comparison with a serologic survey performed in the same region before the CCP disclosed a reduction in seropositive subjects from 47.8% to 17.1% (p less than .001), most marked amongst children and teenagers from 29.9% to 1.9%, suggesting that transmission of the disease had diminished. Similar seropositivity changes after the CCP were observed nationwide. Because decreased superinfection would also be expected to occur, we tried to ascertain whether the clinical outcome of seropositive individuals living in Roscio county had improved. The mean age of seropositive subjects between both surveys increased significantly from 34.9 +/- 17.3 to 46.7 +/- 15.1 years (p less than .001). Additionally, we examined clinically and obtained electrocardiograms from 775 seropositive subjects. They were classified as asymptomatic (group A, n = 614) or as symptomatic, having mild-to-moderate heart symptoms (group B, n = 99) or having advanced congestive heart failure (group C, n = 62). Their electrocardiograms were compared with those of 923 seronegative subjects collected simultaneously and with published data obtained before the CCP. Comparison of the age-related rates of electrocardiographic abnormalities of seropositive individuals before and after the CCP showed that they did not differ significantly by linear regression analysis, by the Kruskal-Wallis test, or by the normal approximation to the binomial distribution.(ABSTRACT TRUNCATED AT 250 WORDS)