Triatomine Research Articles

Assessing the impact of a potential canine vaccine for the control of Chagas disease: a mathematical modeling study

IntroductionChagas disease, a zoonotic infection transmitted by triatomine bugs, poses serious public health risks in endemic areas. As dogs are important reservoirs in the disease’s spread, developing a canine vaccine could be transformative for controlling disease transmission to dogs and humans.MethodsWe developed a compartmental Susceptible-Infected model to simulate the transmission dynamics of Trypanosoma cruzi, considering interactions among dogs, humans, cats, rodents, and triatomine vectors. We used the model to assess the direct and indirect impacts of two vaccine mechanisms—all-or-nothing and leaky—on disease incidence across different host populations. The sensitivity of the model’s outcomes to changes in input parameters was analyzed using univariate sensitivity analysis.ResultsOur model showed that with a 90% vaccine efficacy, an all-or-nothing vaccine could reduce the cumulative incidence of T. cruzi in dogs by 91.3% over five years. The 60% and 30% vaccine efficacies would result in reductions of 63.47% and 33%, respectively, over 5 years. Similarly, the leaky vaccine achieved a 92.62% reduction in dog infections over 5 years with 90% efficacy. The indirect effects on human T. cruzi infection were notable; the all-or-nothing vaccine reduced human disease incidence by 14.37% at 90% efficacy, while the leaky vaccine achieved a 32.15% reduction over 5 years. Both vaccine mechanisms may substantially reduce T. cruzi incidence among dogs, and generate indirect benefit to other hosts, such as humans, by reducing their infection risk. The indirect benefits of vaccination were heavily influenced by the proportion of triatomine bugs blood meals taken from dogs.DiscussionThe study highlights the potential of targeted canine vaccination in controlling Chagas disease transmission and burden in endemic countries. It provides additional evidence for pursuing the development of a canine vaccine as a valuable tool for Chagas disease elimination.

Genome report: First whole genome sequence of Triatoma sanguisuga (Le Conte, 1855), vector of Chagas disease.

Triatoma sanguisuga is the most widespread triatomine bug species in the United States (US). The species vectors the human parasite Trypanosoma cruzi , which causes Chagas disease. Vector-borne Chagas disease is rarely diagnosed in the US, but T. sanguisuga has been implicated in a handful of cases. Despite its public health importance, little is known about the genomics or population genetics of T. sanguisuga . Here, we used long-read sequencing to assemble the first whole genome sequence for T. sanguisuga using DNA extracted from one adult specimen from Delaware. The final size of the genome was 1.162 Gbp with 77.7x coverage. The assembly consisted of 183 contigs with an N50 size of 94.97 Kb. The Benchmarking Universal Single-Copy Ortholog (BUSCO) complete score was 99.1%, suggesting a very complete assembly. Genome-wide GC level was 33.56%, and DNA methylation was 18.84%. The genome consists of 61.4% repetitive DNA and 17,799 predicted coding genes. The assembled T. sanguisuga genome was slightly larger than that of Triatominae species Triatoma infestans and Rhodnius prolixus (949 Mbp with 90.4% BUSCO score and 706 Mbp with 96.5% BUSCO score, respectively). The T. sanguisuga genome is the first North American triatomine species genome to be sequenced, and it is the most complete genome yet for any Triatominae species. The T. sangsuiga genome will allow for deeper investigations into epidemiologically relevant aspects of this important vector species, including blood feeding, host seeking, and parasite competence, thus providing potential vector-borne disease management targets and strengthening public health preparedness.

From Trypomastigotes to Trypomastigotes: Analyzing the One-Way Intracellular Journey of Trypanosoma cruzi by Ultrastructure Expansion Microscopy.

The protozoan parasite Trypanosoma cruzi is the causative agent of Chagas disease, also called American trypanosomiasis. This neglected tropical disease affects millions of individuals across the Americas. To complete its life cycle, T. cruzi parasitizes both vertebrate hosts and its vector, commonly known as the 'kissing bug'. The parasite's survival and proliferation strategies are driven by the diverse environments it encounters. Despite being described by Carlos Chagas in 1909, significant knowledge gaps persist regarding the parasite's various life forms and adaptive capabilities in response to environmental cues. In this study, we employed Ultrastructure Expansion Microscopy to explore the intricate journey of T. cruzi within the host cell. Upon entry into the host cell, trypomastigotes undergo folding, resulting in intermediate forms characterized by a rounded cell body, anterior positioning of basal bodies, and a shortened flagellum. The repositioning of basal bodies and the kinetoplast and the shortening of the flagella mark the culmination of intracellular amastigogenesis. Furthermore, we analyzed intracellular trypomastigogenesis, identifying discrete intermediate forms, including leaf-shaped stages and epimastigote-like forms, which suggests a complex differentiation process. Notably, we did not observe any dividing intracellular epimastigotes, indicating that these may be non-replicative forms within the host cell. Our detailed examination of amastigote cell division revealed semi-closed nuclear mitosis, with mitotic spindle formation independent of basal bodies. This study provides new insights into the morphological and cytoskeletal changes during the intracellular stages of T. cruzi, providing a model for understanding the dynamics of intracellular amastigogenesis and trypomastigogenesis.

Leveraging transcriptome sequence read archives for virus detection in wild and colony populations of triatomines (Hemiptera: Reduviidae: Triatominae)

Triatomines are infamous as vectors of the parasite Trypanosoma cruzi, the causative agent of Chagas disease. However, climate-driven range expansion and urbanization adaptation of triatomine populations, coupled with their highly diverse feeding strategies (vertebrate haematophagy, kleptohaematophagy, and coprophagy), and has elevated interest in triatomines as potential arboviral vectors. Information on the triatomine virome is scant, with prior records including only eight insect-specific viruses: Triatoma virus (TrV) and Rhodnius prolixus viruses 1–7. Here, we leverage publicly available transcriptome datasets to assess viral diversity in 122 wild and colony kissing bugs representing eight species from six countries. In total, six viruses were detected (including Rhodnius prolixus viruses 4–6), and TrV was detected in almost half of all screened triatomines. This is the first report of TrV in Triatoma brasiliensis and in members of the genus Mepraia (M. gajardoi, M. spinolai, and M. parapatrica), and this effort has vastly expanded the publicly available genomic resources of TrV, adding 39 genome sequences to the single genome sequence currently available in the GenBank database. Furthermore, two additional viruses—Meccus longipennis virus 1 and Drosophila melanogaster Nora virus—are herein reported for the first time from kissing bugs. Meccus longipennis virus 1 was detected in Triatoma infestans from Argentina, Brazil, Chile, and Peru, and Drosophila melanogaster Nora virus was found in T. infestans from Argentina. Our results illustrate the advantage and utility of low-cost transcriptome data mining for the discovery of known and novel arboviruses in triatomines and other potential insect vectors.

A baited trap for kissing bugs (Hemiptera: Reduviidae: Triatominae)

Chagas disease is a key vector-borne disease. This illness is caused by Trypanosoma cruzi Chagas, which is transmitted by triatomine bugs. Largely, the control of this disease relies on reducing such contact. We optimized the performance of a box trap in laboratory conditions to capture four triatomine species: Triatoma pallidipennis (Stål), Triatoma infestans Klug, Triatoma phyllosoma (Burmeister), and Rhodnius prolixus Stål. We varied four components for a box trap: material, color, height, and bait attractants. All species were captured more in corrugated cardboard traps than in other trap material. Moreover, T. infestans and R. prolixus were also captured in plywood traps. T. pallidipennis preferred traps of 15 × 15 × 4 cm and 20 × 20 × 4 cm, while T. phyllosoma and T. infestans were more captured in traps of 10 × 10 × 4 cm, and 15 × 15 × 4 cm. Rhodnius prolixus was more captured to 10 × 10 × 4 cm traps. T. pallidipennis was trapped with traps of any color tested, T. phyllosoma and T. infestans were captured more in red and yellow traps, and R. prolixus was mostly captured in blue, violet, and yellow traps. Triatoma pallidipennis was captured at any height above the ground, while T. phyllosoma, T. infestans, and R. prolixus were mostly captured 50, 100, and 150 cm above the ground. Regarding the lure, T. pallidipennis was trapped with four aldehydes + lactic acid + ammonia; T. infestans and R. prolixus were trapped with a blend of four aldehydes + lactic acid, a blend of the four aldehydes + ammonia, and a blend of four aldehydes + lactic acid + ammonia. Triatoma phyllosoma was trapped with any lure tested. These results showed that the trap boxes offer an alternative method for controlling Chagas disease.

Range size positively correlates with temperature and precipitation niche breadths but not with dietary niche breadth in triatomine insects, vectors of Chagas disease.

Ecological theory predicts that species that can utilise a greater diversity of resources and, therefore, have wider niche breadths should also occupy larger geographic areas (the 'niche breadth-range size hypothesis'). Here, we tested this hypothesis for a blood-sucking group of insects of medical significance: the Triatominae (aka 'kissing bugs') (Hemiptera: Reduviidae). Given that niches can be viewed from different perspectives, we tested this hypothesis based on both dietary and climatic niches. We assembled the most complete dataset of triatomine feeding patterns to date by reviewing 143 studies from the literature up to 2021 and tested whether the niche breadth-range size hypothesis held for this group for both dietary and climatic components of the niche. Temperature and precipitation niche breadths were estimated from macro-environmental variables, while diet breadth was calculated based on literature data that used PCR and/or ELISA to identify different types of hosts as blood sources per triatomine species. Our results showed that temperature and precipitation niche breadths, but not dietary breadth, were positively correlated with range sizes, independent of evolutionary history among species. These findings support the predictions from the range size-niche breadth hypothesis concerning climate but not diet, in Triatominae. It also shows that support for the niche breadth-range size hypothesis is dependent upon the niche axis under consideration, which can explain the mixed support for this hypothesis in the ecological literature.

The Rpfor gene modulates the locomotory activity and host-seeking behaviour of Rhodnius prolixus.

The molecular bases of animal behaviour are intricate due to the pleiotropic nature of behaviour-modulating genes, which are often expressed across multiple tissues. The foraging gene (for) encodes a cGMP-dependent protein kinase (PKG), pivotal in regulating downstream target proteins through phosphorylation. In insects, for has been implicated in various behavioural contexts and physiological processes regarding searching for food. Rhodnius prolixus, a hematophagous bug that transmits Trypanosoma cruzi, the causative agent of Chagas disease, exhibits specific activity patterns associated with its hematophagous behaviour. Our previous work demonstrated a correlation between locomotor activity profiles and the expression of Rpfor, suggesting its involvement in modulating triatomine locomotion. In this study, we investigated the impact of Rpfor knockdown on locomotory activity, host-seeking behaviour, feeding performance and lipid metabolism in R. prolixus nymphs. Using RNA interference, we achieved a significant reduction of Rpfor expression in both the brain and fat body of R. prolixus nymphs. Knocked-down nymphs exhibited diminished non-oriented locomotory activity compared with controls, without altering the characteristic bimodal pattern of activity. Additionally, they displayed an increased tendency to approach a host, suggesting a role for Rpfor in modulating host-seeking behaviour. Feeding performance and lipid metabolism remained unaffected by Rpfor knockdown. Our findings underscore the multifaceted role of Rpfor in modulating locomotor activity and host-seeking behaviour in R. prolixus nymphs, shedding light on the molecular mechanisms underlying their hematophagous behaviour and potential implications for disease transmission. Further research is necessary to elucidate the intricate interplay between Rpfor expression, behaviour and physiological processes in triatomine bugs.

Antimicrobial activity of NK cells to Trypanosoma cruzi infected human primary Keratinocytes.

Infection with the protozoan parasite Trypanosoma cruzi is causative for Chagas disease, which is a highly neglected tropical disease prevalent in Latin America. Humans are primary infected through vectorial transmission by blood-sucking triatomine bugs. The parasite enters the human host through mucous membranes or small skin lesions. Since keratinocytes are the predominant cell type in the epidermis, they play a critical role in detecting disruptions in homeostasis and aiding in pathogen elimination by the immune system in the human skin as alternative antigen-presenting cells. Interestingly, keratinocytes also act as a reservoir for T. cruzi, as the skin has been identified as a major site of persistent infection in mice with chronic Chagas disease. Moreover, there are reports of the emergence of T. cruzi amastigote nests in the skin of immunocompromised individuals who are experiencing reactivation of Chagas disease. This observation implies that the skin may serve as a site for persistent parasite presence during chronic human infection too and underscores the significance of investigating the interactions between T. cruzi and skin cells. Consequently, the primary objective of this study was to establish and characterize the infection kinetics in human primary epidermal keratinocytes (hPEK). Our investigation focused on surface molecules that either facilitated or hindered the activation of natural killer (NK) cells, which play a crucial role in controlling the infection. To simulate the in vivo situation in humans, an autologous co-culture model was developed to examine the interactions between T. cruzi infected keratinocytes and NK cells. We evaluated the degranulation, cytokine production, and cytotoxicity of NK cells in response to the infected keratinocytes. We observed a strong activation of NK cells by infected keratinocytes, despite minimal alterations in the expression of activating or inhibitory ligands on NK cell receptors. However, stimulation with recombinant interferon-gamma (IFN-γ), a cytokine known to be present in significant quantities during chronic T. cruzi infections in the host, resulted in a substantial upregulation of these ligands on primary keratinocytes. Overall, our findings suggest the crucial role of NK cells in controlling acute T. cruzi infection in the upper layer of the skin and shed light on keratinocytes as potential initial targets of infection.

Development of an operational trap for collection, killing, and preservation of triatomines (Hemiptera: Reduviidae): the kissing bug kill trap.

Surveillance of triatomines or kissing bugs (Hemiptera: Reduviidae: Triatominae), the insect vectors of Trypanosoma cruzi, a Chagas disease agent, is hindered by the lack of an effective trap. To develop a kissing bug trap, we made iterative improvements over 3 years on a basic design resulting in 7 trap prototypes deployed across field sites in Texas, United States and Northern Mexico, yielding the capture of 325 triatomines of 4 species (Triatoma gerstaeckeri [Stål], T. sanguisuga [LeConte], T. neotomae [Neiva], and T. rubida [Uhler]). We began in 2019 with vertical transparent tarpaulin panel traps illuminated with artificial light powered by AC current, which were successful in autonomous trapping of flying triatomines, but were expensive, labor-intensive, and fragile. In 2020, we switched to white LED lights powered by a solar cell. We tested a scaled-down version of the vertical panel traps, a commercial cross-vane trap, and a multiple-funnel trap. The multiple-funnel traps captured 2.6× more kissing bugs per trap-day than cross-vane traps and approached the performance of the vertical panel traps in number of triatomines captured, number of triatomines per trap-day and triatomines per arthropod bycatch. Multiple-funnel traps required the least labor, were more durable, and had the highest triatomines per day per cost. Propylene glycol in the collection cups effectively preserved captured triatomines allowing for molecular detection of T. cruzi. The trapping experiments established dispersal patterns for the captured species. We conclude that multiple-funnel traps with solar-powered LED lights should be considered for adoption as surveillance and potentially mass-trapping management tools for triatomines.

Life history data of four populations of Triatoma mexicana (Hemiptera: Reduviidae) from Central Mexico.

Triatomine bugs are vectors for the Trypanosoma cruzi Chagas parasites, the etiological agent for Chagas disease. This study evaluated 6 epidemiologically significant behaviors (development time, number of blood meals required for molting to the next instar, mortality rate, aggressiveness, feeding duration, and defecation delay) across 4 populations of Triatoma mexicana Herrich-Schaeffer (Heteroptera: Reduviidae), a major T. cruzi vector in Central Mexico. We collected triatomines from areas characterized by high (HP), medium (MP), medium-high (MHP), and low (LP) prevalence of human T. cruzi infection. The MHP population had the shortest development time, <290 days. Both the HP and MP populations required the most blood meals to molt to the next instar, with a median of 13. Mortality rates varied across all populations, ranging from 44% to 52%. All of the tested populations showed aggressive behavior during feeding. All populations shared similar feeding durations, with most exceeding 13 min and increasing with each instar. Quick defecation, during feeding, immediately after or less than 1 min after feeding, was observed in most nymphs (78%-90%) from the MP and MHP populations and adults (74%-92%) from HP, MP, and MHP populations. Though most parameters suggest a low potential for T. mexicana to transmit T. cruzi, unique feeding and defecation behaviors in 3 populations (excluding the LP group) could elevate their epidemiological importance. These population-specific differences may contribute to the varying prevalence rates of T. cruzi infection in areas where T. mexicana is found.

Zoonotic Cycle of American Trypanosomiasis in an Endemic Region of the Argentine Chaco, Factors That Influenced a Paradigm Shift.

Trypanosoma cruzi, the causative agent of Chagas disease (American trypanosomiasis), is a highly complex zoonosis that is present throughout South America, Central America, and Mexico. The transmission of this disease is influenced by various factors, including human activities like deforestation and land use changes, which may have altered the natural transmission cycles and their connection to the environment. In this study conducted in the Argentine Chaco region, we examined the transmission dynamics of T. cruzi by collecting blood samples from wild and domestic animals, as well as triatomine bugs from human dwellings, across five sites of varying anthropic intervention. Samples were analyzed for T. cruzi infection via qPCR, and we additionally examined triatomines for bloodmeal analysis via NGS amplicon sequencing. Our analysis revealed a 15.3% infection rate among 20 wild species (n = 123) and no T. cruzi presence in 9 species of domestic animals (n = 1359) or collected triatomines via qPCR. Additionally, we found chicken (34.28%), human (21.59%), and goat (19.36%) as the predominant bloodmeal sources across all sites. These findings suggest that anthropic intervention and other variables analyzed may have directly impacted the spillover dynamics of T. cruzi's sylvatic cycle and potentially reduced its prevalence in human habitats.

A simplified molecular tool for detecting the Chagas etiological agent using a vector feces sample in field conditions

Triatomine bugs are vectors of Trypanosoma cruzi, the etiologic agent of Chagas disease in the American continent. Here, we have tested a loop-mediated isothermal amplification (LAMP) test for a direct detection of T. cruzi in feces of Triatoma infestans, the main vector of this parasite in the Southern Cone of America. The analytical evaluation showed positive results with samples of triatomine feces artificially inoculated with DNA from strains of T. cruzi corresponding to each Discrete Typing Units (I-VI), with a sensitivity of up to one parasite per reaction. Conversely, the reaction yielded negative results when tested with DNA from Trypanosoma rangeli and other phylogenetically related and unrelated organisms. In triatomines captured under real field conditions (from urban households), and defined as positive or negative for T. cruzi using the reference microscopy technique, the LAMP test achieved a concordance of 100 %. Our results demonstrate that this LAMP reaction exhibits excellent analytical specificity and sensitivity without interference from the fecal matrix, since all the reactions were conducted without purification steps. This simple molecular diagnostic technique can be easily used by vector control agencies under field conditions.

A Test of Immune Priming in the Kissing Bug Rhodnius pallescens (Hemiptera: Reduviidae) against the Entomopathogenic Fungus Beauveria bassiana (Hypocreales: Cordycipitaceae) in Panama

Abstract The assumption that the invertebrate immune system lacks memory and specificity has changed over time: many studies now indicate that a primary exposure of the host to a pathogen increases its resistance to a subsequent lethal challenge, a phenomenon known as immune priming. One group of insects in which immune priming has been little investigated is the hematophagous triatomine bugs. Herein, we tested the capability of the kissing bug Rhodnius pallescens Barber (Hemiptera: Reduviidae; hereafter kissing bugs), the vector of Chagas disease, to resist entomopathogenic fungi. Laboratory kissing bugs free of Wolbachia and Trypanosoma spp. as well as kissing bugs collected from the wild were used for tests with the entomopathogen Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae). Against laboratory kissing bugs, the fungus remained virulent for 94 d, indicating long-term viability. Kissing bugs collected from the wild that were exposed to a nonlethal dose of the fungus did not show increased survival against a lethal dose compared with controls inoculated with the lethal dose. However, kissing bugs inoculated with a nonlethal dose had higher levels of total phenoloxidase than control kissing bugs. Although the fungus activates the immune system of the kissing bugs, other variables may influence survival in the face of infection. Moreover, the lethality of the same strain was lower against wild kissing bugs, suggesting that the presence of symbionts or parasites influence the fungus–triatome (host) interaction. This work is one of the few studies that have investigated the fungus–host interaction in terms of immune priming in a hematophagous insect of public health importance. Implications are discussed.

Insecticidal activity of fluralaner (Exzolt®) administered to Gallus gallus domesticus against triatomines (Hemiptera, Reduviidae, Triatominae)

BackgroundTriatoma infestans, Triatoma brasiliensis, Triatoma pseudomaculata and Rhodnius prolixus are vectors of Trypanosoma cruzi, the etiological agent of Chagas disease. Chickens serve as an important blood food source for triatomines. This study aimed to assess the insecticidal activity of fluralaner (Exzolt®) administered to chickens against triatomines (R. prolixus, T. infestans, T. brasiliensis and T. pseudomaculata).MethodsTwelve non-breed chickens (Gallus gallus domesticus) were randomized based on weight into three groups: negative control (n = 4); a single dose of 0.5 mg/kg fluralaner (Exzolt®) (n = 4); two doses of 0.5 mg/kg fluralaner (Exzolt®) (n = 4). Nymphs of 3rd, 4th and 5th instars of R. prolixus, T. infestans, T. brasiliensis and T. pseudomaculata (all n = 10) were allowed to feed on chickens before treatment, and at intervals of 1, 7, 14, 21, 28, 35 and 56 days after treatment, with insect mortality determined.ResultsTreatment with two doses of fluralaner showed higher insecticidal efficacy against R. prolixus, T. infestans and T. brasiliensis compared to the single-dose treatment. Similar insecticidal efficacy was observed for T. pseudomaculata for one and two doses of fluralaner. Insecticidal activity of fluralaner (Exzolt®) against triatomine bugs was noted up to 21 and 28 days after treatment with one and two doses of fluralaner, respectively.ConclusionsThe results demonstrate that treatment of chickens with fluralaner (Exzolt®) induces insecticidal activity against triatomines for up to 28 days post-treatment, suggesting its potential use as a control strategy for Chagas disease in endemic areas.Graphical

Trypanosomes and gut microbiota interactions in triatomine bugs and tsetse flies: A vectorial perspective.

Triatomines (kissing bugs) and tsetse flies (genus: Glossina) are natural vectors of Trypanosoma cruzi and Trypanosoma brucei, respectively. T. cruzi is the causative agent of Chagas disease, endemic in Latin America, while T. brucei causes African sleeping sickness disease in sub-Saharan Africa. Both triatomines and tsetse flies are host to a diverse community of gut microbiota that co-exist with the parasites in the gut. Evidence has shown that the gut microbiota of both vectors plays a key role in parasite development and transmission. However, knowledge on the mechanism involved in parasite-microbiota interaction remains limited and scanty. Here, we attempt to analyse Trypanosoma spp. and gut microbiota interactions in tsetse flies and triatomines, with a focus on understanding the possible mechanisms involved by reviewing published articles on the subject. We report that interactions between Trypanosoma spp. and gut microbiota can be both direct and indirect. In direct interactions, the gut microbiota directly affects the parasite via the formation of biofilms and the production of anti-parasitic molecules, while on the other hand, Trypanosoma spp. produces antimicrobial proteins to regulate gut microbiota of the vector. In indirect interactions, the parasite and gut bacteria affect each other through host vector-activated processes such as immunity and metabolism. Although we are beginning to understand how gut microbiota interacts with the Trypanosoma parasites, there is still a need for further studies on functional role of gut microbiota in parasite development to maximize the use of symbiotic bacteria in vector and parasite control.

Machine learning for predicting Chagas disease infection in rural areas of Brazil.

Chagas disease is a severe parasitic illness that is prevalent in Latin America and often goes unaddressed. Early detection and treatment are critical in preventing the progression of the illness and its associated life-threatening complications. In recent years, machine learning algorithms have emerged as powerful tools for disease prediction and diagnosis. In this study, we developed machine learning algorithms to predict the risk of Chagas disease based on five general factors: age, gender, history of living in a mud or wooden house, history of being bitten by a triatomine bug, and family history of Chagas disease. We analyzed data from the Retrovirus Epidemiology Donor Study (REDS) to train five popular machine learning algorithms. The sample comprised 2,006 patients, divided into 75% for training and 25% for testing algorithm performance. We evaluated the model performance using precision, recall, and AUC-ROC metrics. The Adaboost algorithm yielded an AUC-ROC of 0.772, a precision of 0.199, and a recall of 0.612. We simulated the decision boundary using various thresholds and observed that in this dataset a threshold of 0.45 resulted in a 100% recall. This finding suggests that employing such a threshold could potentially save 22.5% of the cost associated with mass testing of Chagas disease. Our findings highlight the potential of applying machine learning to improve the sensitivity and effectiveness of Chagas disease diagnosis and prevention. Furthermore, we emphasize the importance of integrating socio-demographic and environmental factors into neglected disease prediction models to enhance their performance.

First Report of Chagas Disease Vector Species Triatoma sanguisuga (Hemiptera: Reduviidae) Infected with Trypanosoma cruzi in Delaware.

In July and October 2023, two live triatomine bugs were found inside a home in New Castle County, Delaware. The bugs were identified as Triatoma sanguisuga, the most widespread triatomine bug species in the United States. Triatoma sanguisuga is a competent vector of Trypanosoma cruzi, the causative agent of Chagas disease. The two specimens were tested via real-time PCR (qPCR) for infection with T. cruzi, and one of the specimens was positive. Despite T. sanguisuga being endemic to the area, attainment of accurate species identification and T. cruzi testing of the bugs required multiple calls to federal, state, private, and academic institutions over several months. This constitutes the first report of T. sanguisuga infected with T. cruzi in Delaware. In addition, this is the first published report of T. sanguisuga in New Castle County, the northernmost and most densely populated county in Delaware. New Castle County still conforms to the described geographic range of T. sanguisuga, which spans from Texas to the East Coast of the United States. The T. cruzi infection prevalence of the species has not been studied in the northeastern United States, but collections in southern states have found prevalences as high as 60%. The Delaware homeowner's lengthy pursuit of accurate information about the vector highlights the need for more research on this important disease vector in Delaware.

CURRENT STATUS OF METATRANSCRIPTOMIC AND RELATED STUDIES IN HEMATOPHAGOUS DISEASE-TRANSMITTING VECTORS

The incidence of numerous vector-borne diseases (VBDs) has recently increased alarmingly due to various widespread factors, including unplanned urbanization, greater human mobility, environmental changes, vector resistance to insecticides, and evolving pathogens. In this context, the World Health Organization (WHO) has repositioned effective and sustainable vector control as a key approach to prevent and eliminate VBDs. It has been shown that the microbiome influences development, nutrition, and pathogen defense in disease-transmitting vectors such as mosquitoes, sandflies, tsetse flies, triatomine bugs, and ticks. Consequently, understanding the endogenous regulation of vector biology can aid in developing effective approaches for vector control. In this respect, a metatranscriptomic approach analyzes all the expressed RNAs in an environmental sample (meta-RNAs) and can thus reveal how the metabolic activities of the microbiome influence vector biology. This review includes an extensive analysis of available literature on microbial and viral studies for some of the major hematophagous disease-transmitting arthropods, with a focus on studies that used next generation sequencing (NGS) approaches. Since a consensus terminology for these “meta-sequencing analyses” has not yet been established, a definition of these terms is presented here to provide the framework for systematically sorting the available information for each of the VBDs analyzed here to single out metatranscriptomic analyses. Finally, key gaps in knowledge were identified for some of these hematophagous disease-transmitting arthropods which will prove very useful for driving future studies.

TRPA5 encodes a thermosensitive ankyrin ion channel receptor in a triatomine insect

As ectotherms, insects need heat-sensitive receptors to monitor environmental temperatures and facilitate thermoregulation. We show that TRPA5, a class of ankyrin transient receptor potential (TRP) channels absent in dipteran genomes, may function as insect heat receptors. In the triatomine bug Rhodnius prolixus (order: Hemiptera), a vector of Chagas disease, the channel RpTRPA5B displays a uniquely high thermosensitivity, with biophysical determinants including a large channel activation enthalpy change (72kcal/mol), a high temperature coefficient (Q10= 25), and invitro temperature-induced currents from 53°C to 68°C (T0.5= 58.6°C), similar to noxious TRPV receptors in mammals. Monomeric and tetrameric ion channel structure predictions show reliable parallels with fruit fly dTRPA1, with structural uniqueness in ankyrin repeat domains, the channel selectivity filter, and potential TRP functional modulator regions. Overall, the finding of a member of TRPA5 as a temperature-activated receptor illustrates the diversity of insect molecular heat detectors.

Development and survival of Triatoma barberi and Triatoma longipennis (Hemiptera: Reduviidae) is affected by Trypanosoma cruzi colonization

Triatomines are insect vectors of the flagellate protozoan Trypanosoma cruzi, the causative agent of Chagas disease. This occurs when the triatomine defecates on the skin of the vertebrate host when it bites its host for feeding. This lesion causes the penetration of T. cruzi, whose infective phase is the tripomastigote, and the non-infective replicative phase epimastigote. Although the effects of T. cruzi on the development and survival of different species of triatomine bugs are known, it has not been fully described how the parasite affects many of the species that inhabit Mexico. Therefore, the present study aimed to investigate the effect of the presence of T. cruzi on triatomine development and survival. For this, five triatomines of each instar of each species, Triatoma barberi and Triatoma longipennis, were inoculated, from their second instar to their adult stage, female or male. Each triatomine was infected with 3-5 x105 parasites and they were incubated for 100 days and the kinetic of parasites in the feces and T. cruzi phases were recorded. The results showed that T. barberi and T. longipennis development was affected in all stages because of T. cruzi infection. It was greater for T. longipennis, both in its mortality, as well as time to molt in each phase.