Trypanosoma cruzi infection reduces the population fitness of Mepraia spinolai, a Chagas disease vector.

The hematophagous insect Mepraia spinolai (Hemiptera: Reduviidae: Triatominae) is naturally infected with the protozoan parasite Trypanosoma cruzi, the agent of Chagas disease in humans. In this study, we compared the demographic parameters of M. spinolai with and without T. cruzi infection. We collected the immature life table data of 479 M. spinolai individuals of control cohort (reared on mice without T. cruzi infection) and 563 M. spinolai individuals of treatment cohort (reared on mice with T. cruzi infection). Nymphs were maintained in individual compartments inside a growth chamber (26°C; 65-75%) until adult emergence; moulting and survival were recorded daily. For the adult life table study of the control, we used 24 pairs of adults from the control cohort. For the adult life table study of T. cruzi-infected cohort, 25 infected females were paired with 25 males from the control cohort. Life table data were analysed using bootstrap-match technique based on the age-stage, two-sex life table. The preadult survival rate (0.5282) of the control cohort was significantly higher than that of the infected cohort (0.2913). However, the mean fecundity of reproductive females (Fr = 22.29 eggs/♀) and net reproductive rate of population (R0 = 5.07 offspring/individual) of the 0.5th percentile bootstrap-match control cohort were not significantly different from those of the infected cohort (Fr = 23.35 eggs/♀, R0 = 3.77 offspring/individual). Due to the shorter total preoviposition period and higher proportion of reproductive female, the intrinsic rate of increase (r = 0.0053 d-1 ) and finite rate of increase (λ = 1.0053 d-1 ) of control cohort of M. spinolai were significantly higher than those of the T. cruzi-infected cohort (r = 0.0035 d-1 , λ = 1.0035 d-1 ). These results suggest that T. cruzi infection reduces the population fitness of the Chagas disease vector M. spinolai.

Revised New World bioregions and environmental correlates for vectors of Chagas disease (Hemiptera, Triatominae).

The subfamily Triatominae includes a group of hematophagous insects, vectors of the parasite Trypanosoma cruzi, which is the etiological agent of Chagas disease, also known as American trypanosomiasis. Triatomines occur in the Old and New World and occupy diverse habitats including tropical and temperate areas. Some studies suggest the distributions of triatomines group into three or four regions. This study objectively determined bioregions focused specifically on New World Triatominae, using clustering and ordination analysis. We also identified indicator species by bioregion and investigated relationships among bioregions and environmental variables using redundancy analysis and multivariate regression trees. We delineated seven bioregions specific to Triatominae and linked each with indicator species. This result suggests more biogeographical structure exists than was revealed in earlier studies that were more general, subjective, and based on older taxonomic and distributional information. Precipitation, elevation, and vegetation were important variables in the delimitating bioregions. This implies that more detailed study of how these factors influence triatomine distributions could benefit understanding of how Chagas disease is spread.

Acute Chagas Disease Presenting as Preseptal Cellulitis.

Chagas disease, also known as American trypanosomiasis, is caused by Trypanosoma cruzi, a parasite transmitted by hematophagous triatomine insects (subfamily Triatominae) belonging to the Reduviidae family, order Hemiptera. Infection occurs through contact with the feces of the infected vector at the site of its bite or on intact mucosa. [Pediatr Ann. 2023;52(10):e394-e397.].

Insights into the microRNA landscape of Rhodnius prolixus, a vector of Chagas disease.

The growing interest in microRNAs (miRNAs) over recent years has led to their characterization in numerous organisms. However, there is currently a lack of data available on miRNAs from triatomine bugs (Reduviidae: Triatominae), which are the vectors of the protozoan parasite Trypanosoma cruzi, the causative agent of Chagas disease. A comprehensive understanding of the molecular biology of vectors provides new insights into insect-host interactions and insect control approaches, which are key methods to prevent disease incidence in endemic areas. In this work, we describe the miRNome profiles from gut, hemolymph, and salivary gland tissues of the Rhodnius prolixus triatomine. Small RNA sequencing data revealed abundant expression of miRNAs, along with tRNA- and rRNA-derived fragments. Fifty-two mature miRNAs, previously reported in Ecdysozoa, were identified, including 39 ubiquitously expressed in the three tissues. Additionally, 112, 73, and 78 novel miRNAs were predicted in the gut, hemolymph, and salivary glands, respectively. In silico prediction showed that the top eight most highly expressed miRNAs from salivary glands potentially target human blood-expressed genes, suggesting that R. prolixus may modulate the host's gene expression at the bite site. This study provides the first characterization of miRNAs in a Triatominae species, shedding light on the role of these crucial regulatory molecules.

Humans as blood-feeding sources in sylvatic triatomines of Chile unveiled by next-generation sequencing.

BACKGROUND: Triatomines are blood-sucking insects capable of transmitting Trypanosoma cruzi, the parasite that causes Chagas disease in humans. Vectorial transmission entails an infected triatomine feeding on a vertebrate host, release of triatomine infective dejections, and host infection by the entry of parasites through mucous membranes, skin abrasions, or the biting site; therefore, transmission to humans is related to the triatomine-human contact. In this cross-sectional study, we evaluated whether humans were detected in the diet of three sylvatic triatomine species (Mepraia parapatrica, Mepraia spinolai, and Triatoma infestans) present in the semiarid-Mediterranean ecosystem of Chile. METHODS: We used triatomines collected from 32 sites across 1100 km, with an overall T. cruzi infection frequency of 47.1% (N = 4287 total specimens) by conventional PCR or qPCR. First, we amplified the vertebrate cytochrome b gene (cytb) from all DNA samples obtained from triatomine intestinal contents. Then, we sequenced cytb-positive PCR products in pools of 10-20 triatomines each, grouped by site. The filtered sequences were grouped into amplicon sequence variants (ASVs) with a minimum abundance of 100 reads. ASVs were identified by selecting the best BLASTn match against the NCBI nucleotide database. RESULTS: Overall, 16 mammal (including human), 14 bird, and seven reptile species were identified in the diet of sylvatic triatomines. Humans were part of the diet of all analyzed triatomine species, and it was detected in 19 sites representing 12.19% of the sequences. CONCLUSIONS: Sylvatic triatomine species from Chile feed on a variety of vertebrate species; many of them are detected here for the first time in their diet. Our results highlight that the sylvatic triatomine-human contact is noteworthy. Education must be enforced for local inhabitants, workers, and tourists arriving in endemic areas to avoid or minimize the risk of exposure to Chagas disease vectors.

Infection susceptibility and vector competence of Rhodnius robustus Larrousse, 1927 and R. pictipes Stal, 1872 (Hemiptera, Reduviidae, Triatominae) for strains of Trypanosoma cruzi (Chagas, 1909) (Kinetoplastida, Trypanosomatidae) I, II and IV.

BACKGROUND: Rhodnius robustus and Rhodnius pictipes are vectors of Trypanosoma cruzi, the etiologic agent of Chagas disease (CD), that are found in the Brazilian Amazon region. Susceptibility to infection and vector competence depend on the parasite-vector relationship. Our objective was to evaluate the interaction between T. cruzi and these two triatomine vectors in pure and mixed experimental infections of T. cruzi strains from the same or different geographic regions. METHODS: Fifth-instar nymphs of R. robustus and R. pictipes were fed on mice infected with four T. cruzi strains, namely genotypes TcIAM, TcIMG, TcIIPR, and TcIVAM, respectively, from the Brazilian states of Amazonas, Minas Gerais and Paraná. Over a period of 120 days, excreta were examined every 20 days to assess vector competence, and intestinal contents (IC) were examined every 30 days to determine susceptibility to infection. RESULTS: The highest positive rate in the fresh examination (%+FE, 30.0%), the highest number of parasitic forms (PF, n = 1969) and the highest metacyclogenesis rate (%MC, 53.8%) in the excreta were recorded for R. robustus/TcIVAM. Examination of the IC of R. pictipes revealed a higher number of PF in infections with TcIAM (22,680 PF) and TcIIPR (19,845 PF) alone or in association (17,145 PF), as well as a %+FE of 75.0% with TcII, in comparison with the other genotypes. The highest %MC (100%) was recorded for the mixed infections of TcIAM with TcIIPR or TcIVAM in the IC of R. pictipes. CONCLUSIONS: Overall, both species were found to be susceptible to the T. cruzi strains studied. Rhodnius robustus showed vector competence for genotypes TcIVAM and TcIAM+TcIVAM and R. pictipes for TcIAM+TcIVAM and TcIAM+TcIIPR; there was elimination of infective forms as early as at 20 days. Our results suggest that both the genetics of the parasite and its geographic origin influence the susceptibility to infection and vector competence, alone or in association.

Potential distributions of the parasite Trypanosoma cruzi and its vector Dipetalogaster maxima highlight areas at risk of Chagas disease transmission in Baja California Sur, Mexico, under climate change.

Dipetalogaster maxima is a primary vector of Chagas disease in the Cape region of Baja California Sur, Mexico. The geographic distribution of D. maxima is limited to this small region of the Baja California Peninsula in Mexico. Our study aimed to construct the ecological niche models (ENMs) of this understudied vector species and the parasite responsible for Chagas disease (Trypanosoma cruzi). We modelled the ecological niches of both species under current and future climate change projections in 2050 using four Representative Concentration Pathways (RCPs): RCP 2.6, RCP 4.5, RCP 6.0, and RCP 8.5. We also assessed the human population at risk of exposure to D. maxima bites, the hypothesis of ecological niche equivalency and similarity between D. maxima and T. cruzi, and finally the abundance centroid hypothesis. The ENM predicted a higher overlap between both species in the Western and Southern coastal regions of the Baja California Peninsula. The climate change scenarios predicted a Northern shift in the ecological niche of both species. Our findings suggested that the highly tourist destination of Los Cabos is a high-risk zone for Chagas disease circulation. Overall, the study provides valuable data to vector surveillance and control programs.

Different profiles and epidemiological scenarios: past, present and future.

The multiplicity of epidemiological scenarios shown by Chagas Disease, derived from multiple transmission routes of the aetiological agent, occurring on multiple geo-ecobiosocial settings determines the complexity of the disease and reveal the difficulties for its control. From the first description of the link between the parasite, the vector and its domestic habitat and the disease that Carlos Chagas made in 1909, the epidemiological scenarios of the American Trypanosomiasis has shown a dynamic increasing complexity. These scenarios changed with time and geography because of new understandings of the disease from multiple studies, because of policies change at the national and international levels and because human movements brought the parasite and vectors to new geographies. Paradigms that seemed solid at a time were broken down, and we learnt about the global dispersion of Trypanosoma cruzi infection, the multiplicity of transmission routes, that the infection can be cured, and that triatomines are not only a health threat in Latin America. We consider the multiple epidemiological scenarios through the different T. cruzi transmission routes, with or without the participation of a Triatominae vector. We then consider the scenario of regions with vectors without the parasite, to finish with the consideration of future prospects.

Trypanosoma cruzi, beyond the dogma of non-infection in birds.

Trypanosoma cruzi is a protozoan parasite responsible for Chagas disease affecting seven million people. The disease cycle is maintained between Triatominae insects and Mammalia hosts; a refractory effect against infection was noted in birds, but only verified in poultry. This paper presents a new host record for T. cruzi, the American barn-owl (Tyto furcata). Trypanosoma cruzi DTU II molecular evidence was found in heart, intestine, liver, and breast suggesting an established chronic infection based on the parasite DNA presence in multiple organs but absent in spleen, as in the murine model and chronically infected raccoons (Procyon lotor). For birds, the parasite rejection was explained based on the complement and high body temperature, but these mechanisms vary greatly among the members of the avian class. Therefore, there is a need to investigate whether more bird species can become infected, and if T. furcata has a role in disseminating, transmitting and/or maintaining the parasite.

Trypanosoma cruzi infection in the wild Chagas disease vector, Mepraia spinolai: Parasitic load, discrete typing units, and blood meal sources.

BACKGROUND: Mepraia spinolai, a wild vector of Trypanosoma cruzi in Chile, is an abundant triatomine species that is frequently infected by the parasite that causes Chagas disease. The aim of this study was to determine if the parasitic load of T. cruzi in M. spinolai is related to its blood meal source and the infecting DTUs of T. cruzi. METHODS: The vector was captured in rural areas. In the laboratory, DNA was extracted from its abdomen and T. cruzi was quantified using qPCR. Real time PCR assays for four T. cruzi DTUs were performed. Blood meal sources were identified by real-time PCR amplification of vertebrate cytochrome b gene sequences coupled with high resolution melting (HRM). RESULTS: Trypanosoma cruzi was detected in 735 M. spinolai; in 484 we identified one blood meal source, corresponding to human, sylvatic, and domestic species. From these, in 224 we were able to discriminate the infecting DTU. When comparing the parasitic loads between the unique blood meal sources, no significant differences were found, but infections with more than one DTU showed higher parasitic loads than single infections. DTU TcI was detected in a high proportion of the samples. CONCLUSIONS: Higher parasitic loads are related to a greater number of T. cruzi DTUs infecting M. spinolai, and this triatomine seems to have a wide span of vertebrate species in its diet.

The potential risk of enzootic Trypanosoma cruzi transmission inside four training and re-training military battalions (BITER) in Colombia.

BACKGROUND: Colombia's National Army is one of the largest military institutions in the country based on the number of serving members and its presence throughout the country. There have been reports of cases of acute or chronic cases of Chagas disease among active military personnel. These may be the result of military-associated activities performed in jungles and other endemic areas or the consequence of exposure to Trypanosoma cruzi inside military establishments/facilities located in endemic areas. The aim of the present study was to describe the circulation of T. cruzi inside facilities housing four training and re-training battalions [Battalions of Instruction, Training en Re-training (BITERs)] located in municipalities with historical reports of triatomine bugs and Chagas disease cases. An entomological and faunal survey of domestic and sylvatic environments was conducted inside each of these military facilities. METHODS: Infection in working and stray dogs present in each BITER location was determined using serological and molecular tools, and T. cruzi in mammal and triatomine bug samples was determined by PCR assay. The PCR products of the vertebrate 12S rRNA gene were also obtained and subjected to Sanger sequencing to identify blood-feeding sources. Finally, we performed a geospatial analysis to evaluate the coexistence of infected triatomines and mammals with the military personal inside of each BITER installation. RESULTS: In total, 86 specimens were collected: 82 Rhodnius pallescens, two Rhodnius prolixus, one Triatoma dimidiata and one Triatoma maculata. The overall T. cruzi infection rate for R. pallescens and R. prolixus was 56.1 and 100% respectively, while T. dimidiata and T. maculata were not infected. Eight feeding sources were found for the infected triatomines, with opossum and humans being the most frequent sources of feeding (85.7%). Infection was most common in the common opossum Didelphis marsupialis, with infection levels of 77.7%. Sylvatic TcI was the most frequent genotype, found in 80% of triatomines and 75% of D. marsupialis. Of the samples collected from dogs (n = 52), five (9.6%; 95% confidence interval: 3.20-21.03) were seropositive based on two independent tests. Four of these dogs were creole and one was a working dog. The spatial analysis revealed a sympatry between infected vectors and mammals with the military population. CONCLUSIONS: We have shown a potential risk of spillover of sylvatic T. cruzi transmission to humans by oral and vectorial transmission in two BITER installations in Colombia. The results indicate that installations where 100,000 active military personnel carry out training activities should be prioritized for epidemiological surveillance of Chagas disease.

Trypanosoma cruzi infection follow-up in a sylvatic vector of Chagas disease: Comparing early and late stage nymphs.

Chagas disease is caused by Trypanosoma cruzi and transmitted by the triatomine Mepraia spinolai in the southwest of South America. Here, we examined the T. cruzi-infection dynamics of field-caught M. spinolai after laboratory feeding, with a follow-up procedure on bug populations collected in winter and spring of 2017 and 2018. Bugs were analyzed twice to evaluate T. cruzi-infection by PCR assays of urine/fecal samples, the first evaluation right after collection and the second 40 days after the first feeding. We detected bugs with: the first sample positive and second negative (+/-), the first sample negative and second positive (-/+), and with both samples positive or negative (+/+; -/-). Bugs that resulted positive on both occasions were the most frequent, with the exception of those collected in winter 2018. Infection rate in spring was higher than winter only in 2018. Early and late stage nymphs presented similar T. cruzi-infection rates except for winter 2017; therefore, all nymphs may contribute to T. cruzi-transmission to humans. Assessment of infection using two samples represents a realistic way to determine the infection a triatomine can harbor. The underlying mechanism may be that some bugs do not excrete parasites unless they are fed and maintained for some time under environmentally controlled conditions before releasing T. cruzi, which persists in the vector hindgut. We suggest that T. cruzi-infection dynamics regarding the three types of positive-PCR results detected by follow-up represent: residual T. cruzi in the rectal lumen (+/-), colonization of parasites attached to the rectal wall (-/+), and presence of both kinds of flagellates in the hindgut of triatomines (+/+). We suggest residual T. cruzi-infections are released after feeding, and result 60-90 days after infection persisting in the rectal lumen after a fasting event, a phenomenon that might vary between contrasting seasons and years.

TriatoScore: an entomological-risk score for Chagas disease vector control-surveillance.

BACKGROUND: Triatomine bugs transmit Chagas disease across Latin America, where vector control-surveillance is increasingly decentralized. Locally run systems often deal with highly diverse native-vector faunas-plus, in some areas, domestic populations of non-native species. Flexible entomological-risk indicators that cover native and non-native vectors and can support local decision-making are therefore needed. METHODS: We present a local-scale entomological-risk score ("TriatoScore") that leverages and builds upon information on the ecology-behavior and distribution-biogeography of individual triatomine bug species. We illustrate our approach by calculating TriatoScores for the 417 municipalities of Bahia state, Brazil. For this, we (i) listed all triatomine bug species recorded statewide; (ii) derived a "species relevance score" reflecting whether each species is native/non-native and, if native, whether/how often it invades/colonizes dwellings; (iii) mapped each species' presence by municipality; (iv) for native vectors, weighted presence by the proportion of municipal territory within ecoregions occupied by each species; (v) multiplied "species relevance score" × "weighted presence" to get species-specific "weighted scores"; and (vi) summed "weighted scores" across species to get municipal TriatoScores. Using standardized TriatoScores, we then grouped municipalities into high/moderate/low entomological-risk strata. RESULTS: TriatoScores were higher in municipalities dominated by dry-to-semiarid ecoregions than in those dominated by savanna-grassland or, especially, moist-forest ecoregions. Bahia's native triatomines can maintain high to moderate risk of vector-borne Chagas disease in 318 (76.3%) municipalities. Historical elimination of Triatoma infestans from 125 municipalities reduced TriatoScores by ~ 27% (range, 20-44%); eight municipalities reported T. infestans since Bahia was certified free of Trypanosoma cruzi transmission by this non-native species. Entomological-risk strata based on TriatoScores agreed well with Bahia's official disease-risk strata, but TriatoScores suggest that the official classification likely underestimates risk in 42 municipalities. Of 152 municipalities failing to report triatomines in 2006-2019, two and 71 had TriatoScores corresponding to, respectively, high and moderate entomological risk. CONCLUSIONS: TriatoScore can help control-surveillance managers to flexibly assess and stratify the entomological risk of Chagas disease at operationally relevant scales. Integrating eco-epidemiological, demographic, socioeconomic, or operational data (on, e.g., local-scale dwelling-infestation or vector-infection frequencies, land-use change and urbanization, housing conditions, poverty, or the functioning of control-surveillance systems) is also straightforward. TriatoScore may thus become a useful addition to the triatomine bug control-surveillance toolbox.

Modification of the Daily Activity Pattern of the Diurnal Triatomine Mepraia spinolai (Hemiptera: Reduviidae) Induced by Trypanosoma cruzi (Trypanosomatida: Trypanosomatidae) Infection.

Mepraia spinolai, (Porter) 1934, is a diurnal triatomine endemic to Chile and a wild vector of the protozoan Trypanosoma cruzi, (Chagas) 1909, which causes Chagas disease. Behavioral changes in M. spinolai induced by this parasite have been reported previously, which include detection of a potential host, defecation latency, and some life history traits. In this study we assessed changes in locomotor and daily activity due to infection with T. cruzi. No difference was detected in distance traveled between infected and uninfected individuals. However, the groups differed in their daily activity patterns; infected individuals showed significant reduction of movements during the light phase and concentrated their activity in the dark phase. Uninfected individuals showed no differences in locomotor activity between the phases. The results suggest that T. cruzi induces a displacement in the activity of M. spinolai toward the dark phase of the circadian cycle, which may improve its vector competence.

Diversity and interactions among triatomine bugs, their blood feeding sources, gut microbiota and Trypanosoma cruzi in the Sierra Nevada de Santa Marta in Colombia.

Chagas disease remains a major neglected disease in Colombia. We aimed to characterize Trypanosoma cruzi transmission networks in the Sierra Nevada de Santa Marta (SNSM) region, to shed light on disease ecology and help optimize control strategies. Triatomines were collected in rural communities and analyzed for blood feeding sources, parasite diversity and gut microbiota composition through a metagenomic and deep sequencing approach. Triatoma dimidiata predominated, followed by Rhodnius prolixus, Triatoma maculata, Rhodnius pallescens, Panstrongylus geniculatus and Eratyrus cuspidatus. Twenty-two species were identified as blood sources, resulting in an integrated transmission network with extensive connectivity among sylvatic and domestic host species. Only TcI parasites were detected, predominantly from TcIb but TcIa was also reported. The close relatedness of T. cruzi strains further supported the lack of separate transmission cycles according to habitats or triatomine species. Triatomine microbiota varied according to species, developmental stage and T. cruzi infection. Bacterial families correlated with the presence/absence of T. cruzi were identified. In conclusion, we identified a domestic transmission cycle encompassing multiple vector species and tightly connected with sylvatic hosts in the SNSM region, rather than an isolated domestic transmission cycle. Therefore, integrated interventions targeting all vector species and their contact with humans should be considered.

Machine-learning model led design to experimentally test species thermal limits: The case of kissing bugs (Triatominae).

Species Distribution Modelling (SDM) determines habitat suitability of a species across geographic areas using macro-climatic variables; however, micro-habitats can buffer or exacerbate the influence of macro-climatic variables, requiring links between physiology and species persistence. Experimental approaches linking species physiology to micro-climate are complex, time consuming and expensive. E.g., what combination of exposure time and temperature is important for a species thermal tolerance is difficult to judge a priori. We tackled this problem using an active learning approach that utilized machine learning methods to guide thermal tolerance experimental design for three kissing-bug species: Triatoma infestans, Rhodnius prolixus, and Panstrongylus megistus (Hemiptera: Reduviidae: Triatominae), vectors of the parasite causing Chagas disease. As with other pathogen vectors, triatomines are well known to utilize micro-habitats and the associated shift in microclimate to enhance survival. Using a limited literature-collected dataset, our approach showed that temperature followed by exposure time were the strongest predictors of mortality; species played a minor role, and life stage was the least important. Further, we identified complex but biologically plausible nonlinear interactions between temperature and exposure time in shaping mortality, together setting the potential thermal limits of triatomines. The results from this data led to the design of new experiments with laboratory results that produced novel insights of the effects of temperature and exposure for the triatomines. These results, in turn, can be used to better model micro-climatic envelope for the species. Here we demonstrate the power of an active learning approach to explore experimental space to design laboratory studies testing species thermal limits. Our analytical pipeline can be easily adapted to other systems and we provide code to allow practitioners to perform similar analyses. Not only does our approach have the potential to save time and money: it can also increase our understanding of the links between species physiology and climate, a topic of increasing ecological importance.

Metacyclogenesis of Trypanosoma cruzi in B. ferroae (Reduviidae: Triatominae) and feces infectivity under laboratory conditions / Metaciclogénesis de Trypanosoma cruzi en Belminus ferroae (Reduviidae: Triatominae) y capacidad infectiva de las heces en condiciones de laboratorio.

Introduction: Belminus ferroae is a triatominae with entomophagous behavior. However, it may occasionally feed on vertebrates. Currently, there is no evidence of natural infection with Trypanosoma cruzi or the occurrence of metacyclogenesis in this species. Objective: To test T. cruzi metacyclogenesis in B. ferroae and the infectivity of their feces or intestinal contents in rodents under laboratory conditions. Materials and methods: Twenty nymphs of B. ferroae were infected with an autochthonous strain of T. cruzi (M/HOM/VE/09/P6). Fecal and urine samples were collected from spontaneous droppings or by compressing the bugs' abdomens and, eventually, by removing their gut contents, and then examined at 10, 20, 30, 40, 50, and 60 days. We quantified T. cruzi parasitic load, as well as the evolutionary forms in feces, urine, and intestinal contents by Giemsa staining. Similarly, we evaluated the infectivity of T. cruzi metacyclic trypomastigotes in albino mice. Results: The parasitological analysis showed three insects (15%) infected with T. cruzi at 30 (n=1), 40 (n=1), and 50 (n=1) days post-infection. We observed parasitic loads of up to 1.62 x 105 trypanosomes/mm3 and metacyclogenesis percentages between 3.5% and 6.78%. Conclusions: This is the first time that T. cruzi metacyclogenesis is reported in a species of the genus Belminus under laboratory conditions and the infectivity of Belminus' feces is demonstrated on a vertebrate host.

Introducción. Belminus ferroae es un triatomino de comportamiento entomófago, sin embargo, puede alimentarse de vertebrados ocasionalmente. No se ha demostrado infección natural por Trypanosoma cruzi en esta especie, como tampoco la metaciclogénesis del parásito. Objetivo. Examinar la metaciclogénesis de T. cruzi en B. ferroae y la capacidad infectiva de las heces o sus contenidos intestinales en roedores. Materiales y métodos. Se analizaron las heces y la orina expulsadas espontáneamente por los insectos o mediante compresión abdominal o extracción del contenido intestinal a los 10, 20, 30, 40, 50 y 60 días. Se cuantificó la carga parasitaria de T. cruzi y sus formas evolutivas se identificaron con tinción de Giemsa. Asimismo, se evaluó en ratones albinos la apacidad infectiva de los tripomastigotes metacíclicos de T. cruzi obtenidos de las heces o contenidos intestinales de los especímenes infectados. Resultados. El análisis parasitológico reveló tres (15 %) insectos infectados con T. cruzi a los 30 (n=1), 40 (n=1) y 50 (n=1) días después de la infección con cargas parasitarias de hasta 1,62 x 105 tripanosomas/mm3 y porcentajes de metaciclogénesis entre el 3,5 y el 6,78 %. Conclusiones. Se demuestra por primera vez, en una especie del género Belminus, la metaciclogenésis de T. cruzi en condiciones de laboratorio y la capacidad infectiva de las heces para un huésped vertebrado.

Bioecological aspects of triatomines and marsupials as wild Trypanosoma cruzi reservoirs in urban, peri-urban and rural areas in the Western Brazilian Amazon.

In the Amazon region, Trypanosoma cruzi transmission cycles involve a great diversity of Triatominae vectors and mammal reservoirs. Some Rhodnius spp. mainly inhabit palm trees that act as microhabitats for hosts and vectors. The current study aimed to describe aspects of the bio-ecology of the vectors and reservoirs of T. cruzi in relation to human populations resident near areas with large quantities of palm trees, in rural, peri-urban and urban collection environments, located in the Western Brazilian Amazon. Rhodnius pictipes and Didelphis marsupialis were respectively the most predominant vector and reservoir, with rates of 71% for R. pictipes and 96.5% for D. marsupialis. The vast majority of T. cruzi isolates clustered with TcI. The most prevalent haplotype was TcI COII1 (69.7%). Mauritia flexuosa and Attalea phalerata were the main ecological indicators of infestation by triatomines. Birds were the most common food source (27,71%). T. cruzi isolated from R. robustus has the haplotype HUM-13, previously detected in a chronic Chagas patient living in the same area. Our results demonstrate the relevance of this study, with the occurrence of elevated infection rates in animals, and suggest the importance of the Amazon zones where there is a risk of infection in humans.

Susceptibility dynamics between five Trypanosoma cruzi strains and three triatomine (Hemiptera: Reduviidae) species.

American trypanosomiasis is a zoonosis caused by the parasite Trypanosoma cruzi and is transmitted mainly by blood-sucking insects belonging to the subfamily Triatominae. The importance of this parasite lies in its wide geographical distribution, high morbidity, and the fact that there has not yet been an effective treatment or vaccine. Previous studies have detailed the interactions between different triatomine species and T. cruzi strains. However, the factors necessary to establish infection in triatomines have not yet been fully elucidated. Furthermore, it is postulated that the coexistence between the parasite and triatomines could modulate the susceptibility to infection in these insects. Accordingly, in this study, we evaluated the susceptibility to T. cruzi infection in the species Triatoma (Meccus) pallidipennis, Triatoma barberi, and Triatoma lecticularia, which were infected with Ninoa, H8, INC-5, Sontecomapan, and Hueypoxtla strains. The criteria used to establish susceptibility were the amount of blood ingested by the insects, percentage of infected triatomines, concentration of parasites in feces, and percentage of metacyclic trypomastigotes in feces. These parameters were analyzed by fresh examination and differential count with Giemsa-stained smears. Our main findings suggest the following order of susceptibility concerning infection with T. cruzi: T. lecticularia > T. barberi > T. (Meccus) pallidipennis. Furthermore, the study concludes that an increased susceptibility to infection of triatomines that share the same geographic region with different strains of T. cruzi is not always a fact.

Detection of Trypanosoma cruzi DNA in false negative samples of collected triatomines, xenodiagnosis material, and biopsies of experimentally infected animals.

Direct test over the gut material from triatomine vectors and xenodiagnosis over mammalian hosts are classical techniques for Trypanosoma cruzi parasitological diagnosis. Nevertheless, negative results can be a source of uncertainty. Experimental models have allowed evaluating the tissue invasion of different strains of T. cruzi, but conventional techniques for tissue biopsies involve time-consuming and elaborated procedures and have low sensitivity. Gut material of collected triatomines (microscopically negative) (n = 114), material of mammal xenodiagnoses (microscopically negative) (n = 138), and biopsy material (microscopically negative) from experimentally infected animals (n = 34) with isolates from endemic areas of Chagas' disease from Venezuela were used for DNA extraction and PCR for the amplification of kinetoplast DNA (kDNA) and satellite DNA (sDNA) of T. cruzi. Positive PCR was observed in 53.6% of collected triatomine material, 15.8% of parasitological negative xenodiagnosis material, and 70.6% in biopsies, revealing underestimation by the parasitological tests and the valour of this analysis with preserved material. Anzoátegui was the state with the highest percentage of infection, and the triatomine species Rhodnius prolixus and Panstrongylus geniculatus had the highest percentages of infection. Didelphis marsupialis and Canis familiaris were the most infected by T. cruzi revealed by PCR of xenodiagnosis material. In addition, the PCR technique allowed demonstrating the invasion of T. cruzi in all tissues analyzed, constituting a molecular marker of tissue invasion.