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.

RhoprCAPA-2 acts as a gonadotropin regulating reproduction in adult female, Rhodnius prolixus.

CAPA peptides play diverse roles in insects, modulating muscle contraction, regulating fluid balance, and reproduction. In Rhodnius prolixus, a hematophagous insect and a vector for human Chagas disease, three CAPA peptides are encoded by the capability gene, including RhoprCAPA-1, RhoprCAPA-2, and RhoprCAPA-PK-1. RhoprCAPA-2 is an anti-diuretic hormone in R. prolixus. Here, we explore the involvement of RhoprCAPA-2 in reproduction in adult female R. prolixus. Double-label immunohistochemistry reveals co-localization of RhoprCAPA-2-like and the glycoprotein hormone (GPA2/GPB5) subunit GPB5-like immunoreactivity in neurosecretory cells in the mesothoracic ganglionic mass and in their neurohemal sites, suggesting these peptides can be co-released to regulate physiological processes. qPCR analysis reveals changes in transcript expression levels of the RhoprCAPA receptor (CAPAR) in the fat body and reproductive tissues after feeding in adult female R. prolixus. RNA interference-mediated knockdown of CAPAR transcript decreases egg production and reduces hatching rate and survival rate in female R. prolixus. Downregulation of CAPAR decreases vitellogenin RhoprVg1 transcript expression in the fat body and deceases its receptor RhoprVgR transcript level in the ovaries; accompanied by a reduction in vitellogenin content in the fat body and hemolymph. Incubation of fat body and ovaries in vitro with RhoprCAPA-2 increases RhoprVg1 transcript expression in the fat body, vitellogenin content in the fat body culture medium, and increases RhoprVgR transcript in the ovaries. These findings implicate the CAPA signaling pathway in reproduction, with RhoprCAPA-2 acting as a gonadotropin in adult female R. prolixus.

Comparative analysis of detoxification-related gene superfamilies across five hemipteran species.

BACKGROUND: Hemiptera is one of the most speciose orders of insects, and the most speciose considering Hemimetabola. Through their evolutive history, hemipterans with different feeding habits have adapted to deal with different chemical challenges. Three major gene families are involved in xenobiotic detoxification in insects: the cytochromes P450 (CYPs), carboxyl/cholinesterases (CCEs), and glutathione transferases (GSTs). Here we perform a comparative analysis on the complement of these gene superfamilies across five hemipteran species; four heteropterans (the pentatomid plant feeders Nezara viridula and Halyomorpha halys; the hematophagous Cimex lectularius, Cimicidae, and Rhodnius prolixus, Reduviidae), and one Auchenorrhyncha plant feeder (Nilaparvata lugens). RESULTS: Our results point to an expansion of several enzyme families associated with xenobiotic detoxification in heteropterans with respect to other species and the existence of a dynamic evolution pattern including CYP3 clan, hormone and pheromone processing class in the CCE superfamily, and sigma class in GST superfamily. Other detoxification-related families are reduced in the hemipteran species analyzed here: reduction or even absence of epsilon class and reduced delta class in GST superfamily; absence of mitochondrial CYP12 family; absence of CYP9 family in CYP3 clan; and reduction or even absence of some dietary/detoxification groups of CCEs. Interestingly, the most polyphagous species analyzed here (H. halys) is also the one that presents the largest repertoire of detoxification enzymes. Gene cluster analysis suggests that this could be due to gene duplication events. CONCLUSIONS: The evolutionary analysis performed here reveals characteristics that are both common and particular for heteropterans. The composition and organization of detoxification-related gene families could shed light on evolutionary forces that shaped their divergence. These families are important for both the detoxification of diet products and for conferring tolerance or resistance to synthetic insecticides. Furthermore, we present the first comprehensive analysis of detoxification gene superfamilies in N. viridula, an understudied species in spite of its economic relevance as a crop pest. The information obtained is of interest for basic insect science as well as for the control of harmful species and the management of insecticide resistance.

Effect of salivary CYP4EM1 and CYP4EM2 gene silencing on the life span of Chagas disease vector Rhodnius prolixus (Hemiptera, Reduviidae) exposed to sublethal dose of deltamethrin.

Control of Chagas disease in endemic countries is primarily accomplished through insecticide spraying for triatomine vectors. In this context, pyrethroids are the first-choice insecticide, and the evolution of insect resistance to these insecticides may represent an important barrier to triatomine control. In insects, cytochrome P450s are enzymes involved in the metabolism of xenobiotics and endogenous chemicals that are encoded by genes divided into different families. In this work, we evaluated the role of three Rhodnius prolixus CYP4EM subfamily genes during blood meal and after deltamethrin exposure. CYP4 gene members were expressed in different insect organs (integument, salivary glands (SGs), midgut, fat body and malpighian tubules) at distinct transcriptional levels. CYP4EM1 gene was highly expressed in the SG and was clearly modulated after insect blood meal. Injection of CYP4EM1dsRNA promoted significant reduction in mRNA levels of both CYP4EM1 and CYP4EM2 genes and induced deleterious effects in R. prolixus nymphs subsequently exposed to sublethal doses of deltamethrin (3.4 or 3.8 ng/nymph treated). The higher dose reduced the survival over time and increased susceptibility of R. prolixus nymphs to deltamethrin. A better understanding of this mechanism can help in developing of more efficient strategies to reduce Trypanosoma cruzi vector transmission in Americas.

Acute Chagas Disease Manifesting as Orbital Cellulitis, Texas, USA.

We report a case of acute, vectorborne Chagas disease, acquired locally in central Texas, USA, manifesting as Romaña's sign, which was initially mistaken for orbital cellulitis. After the infection failed to respond to antibiotics, DNA-based next generation sequencing on plasma yielded high levels of Trypanasoma cruzi; results were confirmed by PCR.

Influence of Serratia marcescens and Rhodococcus rhodnii on the Humoral Immunity of Rhodnius prolixus.

Chagas disease is a human infectious disease caused by Trypanosoma cruzi and can be transmitted by triatomine vectors, such as Rhodnius prolixus. One limiting factor for T. cruzi development is the composition of the bacterial gut microbiota in the triatomine. Herein, we analyzed the humoral immune responses of R. prolixus nymphs treated with antibiotics and subsequently recolonized with either Serratia marcescens or Rhodococcus rhodnii. The treatment with antibiotics reduced the bacterial load in the digestive tract, and the recolonization with each bacterium was successfully detected seven days after treatment. The antibiotic-treated insects, recolonized with S. marcescens, presented reduced antibacterial activity against Staphylococcus aureus and phenoloxidase activity in hemolymph, and lower nitric oxide synthase (NOS) and higher defensin C gene (DefC) gene expression in the fat body. These insects also presented a higher expression of DefC, lower prolixicin (Prol), and lower NOS levels in the anterior midgut. However, the antibiotic-treated insects recolonized with R. rhodnii had increased antibacterial activity against Escherichia coli and lower activity against S. aureus, higher phenoloxidase activity in hemolymph, and lower NOS expression in the fat body. In the anterior midgut, these insects presented higher NOS, defensin A (DefA) and DefC expression, and lower Prol expression. The R. prolixus immune modulation by these two bacteria was observed not only in the midgut, but also systemically in the fat body, and may be crucial for the development and transmission of the parasites Trypanosoma cruzi and Trypanosoma rangeli.

Chagas Disease in the United States: a Public Health Approach.

Trypanosoma cruzi is the etiological agent of Chagas disease, usually transmitted by triatomine vectors. An estimated 20 to 30% of infected individuals develop potentially lethal cardiac or gastrointestinal disease. Sylvatic transmission cycles exist in the southern United States, involving 11 triatomine vector species and infected mammals such as rodents, opossums, and dogs. Nevertheless, imported chronic T. cruzi infections in migrants from Latin America vastly outnumber locally acquired human cases. Benznidazole is now FDA approved, and clinical and public health efforts are under way by researchers and health departments in a number of states. Making progress will require efforts to improve awareness among providers and patients, data on diagnostic test performance and expanded availability of confirmatory testing, and evidence-based strategies to improve access to appropriate management of Chagas disease in the United States.

Decoys and Dilution: The Impact of Incompetent Hosts on Prevalence of Chagas Disease.

Biodiversity is commonly believed to reduce risk of vector-borne zoonoses. However, researchers already showed that the effect of biodiversity on disease transmission is not that straightforward. This study focuses on the effect of biodiversity, specifically on the effect of the decoy process (additional hosts distracting vectors from their focal host), on reducing infections of vector-borne diseases in humans. Here, we consider the specific case of Chagas disease and use mathematical population models to observe the impact on human infection of the proximity of chickens, which are incompetent hosts for the parasite but serve as a preferred food source for vectors. We consider three cases as the distance between the two host populations varies: short (when farmers bring chickens inside the home to protect them from predators), intermediate (close enough for vectors with one host to detect the presence of the other host type), and far (separate enclosed buildings such as a home and hen-house). Our analysis shows that the presence of chickens reduces parasite prevalence in humans only at an intermediate distance under the condition that the vector birth rate from feeding on chickens is sufficiently low.

Participation of Trypanosoma cruzi gp63 molecules on the interaction with Rhodnius prolixus.

Trypanosoma cruzi is the causative agent of Chagas disease, a vector-borne disease. The parasite molecules involved in vector interaction have been little investigated. Metallopeptidases and gp63 molecules have been implicated in parasite adhesion of several trypanosomatids to the insect midgut. Although gp63 homologues are highly expanded in the T. cruzi genome, and are implicated in parasite-mammalian host interaction, its role in the insect vector has never been explored. Here, we showed that divalent metal chelators or anti-Tcgp63-I antibodies impaired T. cruzi adhesion to Rhodnius prolixus midgut. Parasites isolated after insect colonization presented a drastic enhancement in the expression of Tcgp63-I. These data highlight, for the first time, that Tcgp63-I and Zn-dependent enzymes contribute to the interaction of T. cruzi with the insect vector.

Synthesis and secretion of volatile short-chain fatty acids in Triatoma infestans infected with Beauveria bassiana.

Physically disturbed Triatoma infestans (Hemiptera: Reduviidae) adults, as well as adults of other Chagas' disease vectors, secrete a mix of volatile organic compounds (VOCs) with alarm and possible sexual and defence functions. The aim of the present research was to test whether infection with the entomopathogenic fungus Beauveria bassiana (Ascomycota: Hypocreales: Clavicipitaceae) has an effect on VOC secretion in disturbed T. infestans and on the expression of two genes (Ti-brnq and Ti-bckdc) potentially involved in VOC biosynthesis. The volatiles released by insects at different time periods after fungal treatment were identified and their relative amounts measured. Isobutyric acid was the most abundant volatile found in both healthy and fungus-infected insects and underwent no significant relative changes through the infection process. The secretion of propionic acid, however, was significantly higher at 1-4 days post-infection (d.p. i.) compared with that in controls. A slight induction of both Ti-brnq and Ti-bckdc genes was found by real-time polymerase chain reaction at 4 d.p. i., with expression values reaching up to three-fold those in controls. The early stages of fungal infection seem to affect the composition of the alarm pheromone by changing the expression pattern of both genes analysed. These results help to elucidate the impact of fungal infections on the chemical ecology of triatomine bugs.

The involvement of Rhopr-CRF/DH in feeding and reproduction in the blood-gorging insect Rhodnius prolixus.

Rhodnius prolixus is a blood-gorging insect and a vector for human Chagas disease. The insect transmits the disease following feeding, when it excretes urine and feces contaminated with the Trypanosoma cruzi parasite. A corticotropin-releasing factor-like peptide acts as a diuretic hormone in R. prolixus (Rhopr-CRF/DH); however, its distribution throughout the insect's central nervous system (CNS) and the expression of its receptor in feeding-related tissue as well as the female reproductive system suggests a multifaceted role for the hormone beyond that of diuresis. Here we investigate the involvement of Rhopr-CRF/DH in feeding and reproduction in R. prolixus. Immunohistochemistry of the CNS showed diminished CRF-like staining in neurosecretory cells (NSCs) of the mesothoracic ganglionic mass (MTGM) immediately following feeding, and partial restocking of those same cells two hours later, indicating Rhopr-CRF/DH stores in this regions are involved in feeding. The results of the temporal qPCR analysis were consistent with the immunohistochemical findings, showing an increase in Rhopr-CRF/DH transcript expression in the MTGM immediately after feeding, presumably capturing the restocking of Rhopr-CRF/DH in the lateral NSCs following release of the peptide during feeding. Elevating haemolymph Rhopr-CRF/DH titres by injection of Rhopr-CRF/DH prior to feeding resulted in the intake of a significantly smaller blood meal in 5th instars and adults without an apparent effect on the rate of short-term diuresis. When adult females were injected with Rhopr-CRF/DH, they also produced and laid significantly fewer eggs. Finally, in vitro oviduct contraction assays illustrate that Rhopr-CRF/DH inhibits the amplitude of contractions of the lateral oviducts, highlighting a potential mechanism via which the hormone diminishes reproductive capacity. To conclude, the study of the Rhopr-CRF/DH pathway, its components and mechanisms of action, has implications for vector control by highlighting targets to alter feeding, diuresis, and reproduction of this disease vector.

Patterns of vector species richness and species composition as drivers of Chagas disease occurrence in Brazil.

Chagas disease represents one of the major health issue in Latin America. Epidemiological control is focused on disease vectors, so studies on the ecology of triatomine vectors constitute a central strategy. Recently, research at large spatial scale has been produced, and authors commonly rely on the assumption that geographical regions presenting good environmental conditions for most vector species are also those with high risk of infection. In the present work, we provide an explicit evaluation for this assumption. Employing species distribution models and epidemiological data for Chagas disease in Brazilian territory, our results show that species richness is a poor predictor for the observed pattern of Chagas disease occurrence. Species composition proved to be a better predictor. We stress that research on macroecology of infectious diseases should go beyond the analysis of biodiversity patterns and consider human infections as a central part of the focal ecological systems.

Hosts and vectors of Trypanosoma cruzi discrete typing units in the Chagas disease endemic region of the Paraguayan Chaco.

Active Trypanosoma cruzi transmission persists in the Gran Chaco region, which is considered hyperendemic for Chagas disease. Understanding domestic and sylvatic transmission cycles and therefore the relationship between vectors and mammalian hosts is crucial to designing and implementing improved effective control strategies. Here we describe the species of triatomine vectors and the sylvatic mammal reservoirs of T. cruzi, in different localities of the Paraguayan and Bolivian Chaco. We identify the T. cruzi genotypes discrete typing units (DTUs) and provide a map of their geographical distribution. A total of 1044 triatomines and 138 sylvatic mammals were captured. Five per cent of the triatomines were microscopically positive for T. cruzi (55 Triatoma infestans from Paraguay and one sylvatic Triatoma guasayana from Bolivia) and 17 animals (12·3%) comprising eight of 28 (28·5%) Dasypus novemcinctus, four of 27 (14·8%) Euphractus sexcinctus, three of 64 (4·7%) Chaetophractus spp. and two of 14 (14·3%) Didelphis albiventris. The most common DTU infecting domestic triatomine bugs was TcV (64%), followed by TcVI (28%), TcII (6·5%) and TcIII (1·5%). TcIII was overwhelmingly associated with armadillo species. We confirm the primary role of T. infestans in domestic transmission, armadillo species as the principal sylvatic hosts of TcIII, and consider the potential risk of TcIII as an agent of Chagas disease in the Chaco.

Selective Insecticide Applications Directed Against Triatoma infestans (Hemiptera: Reduviidae) Affected a Nontarget Secondary Vector of Chagas Disease, Triatoma garciabesi.

The control of nondomiciliated triatomine species adapted to peridomestic habitats represents a challenge because they are connected to sylvatic colonies, and pyrethroid insecticides have limited effects outdoors. The effects of residual insecticide spraying have rarely been assessed on secondary triatomines. Triatoma garciabesi (Carcavallo, Martinez, Cichero, Prosen & Ronderos, 1967) is a nontarget vector that inhabits the dry western Chaco region, and a member of the Triatoma sordida Stål 1859 complex. Little is known on the capacity of T. garciabesi to invade and establish viable domestic or peridomestic colonies, and on its response to residual insecticide sprays directed against Triatoma infestans Klug 1834. The presence and abundance of triatomines were assessed by timed manual collections annually or biannually (spring and fall) during 10 yr after a community-wide insecticide spraying campaign and selective insecticide sprays directed against T. infestans in a rural village of northwestern Argentina. T. garciabesi mainly occupied peridomestic habitats associated with chickens, and was unable to colonize human sleeping quarters. Trees with chickens occurred in nearly all houses and were infested in >25% of the occasions. The abundance of bugs at house-compound level was best explained by a generalized estimating equation model that included selective insecticide sprays during the previous semester (negative effects), chicken abundance (positive effects), seasonality, and their interactions. Our results suggest that insecticide applications targeting T. infestans affected the abundance of T. garciabesi, and reduced the likelihood of future infestation.

Infection Rate by Trypanosoma cruzi and Biased Vertebrate Host Selection in the Triatoma dimidiata (Hemiptera: Reduvidae) Species Complex.

Chagas disease is a vector-borne disease, caused by the protozoan parasite Trypanosoma cruzi and transmitted by hematophagous insects. Triatoma dimidiata (Hemiptera: Reduvidae (Latreille 1811)) is one of the main vectors, and recent molecular studies indicate that it is a species complex, with potentially different vectorial competences. We investigated the differences in natural T. cruzi infection rate within T. dimidiata complex in Yucatan, Mexico. ITS-2 hybrid bugs had a twofold higher infection rate than ITS-2 Groups 2 and 3 bugs, and this pattern was consistent over time and in several villages. To test if T. dimidiata ITS-2 hybrid bugs could feed more frequently on T. cruzi-infected hosts, we evaluated their host-seeking behavior in a dual-choice chamber. Group 2 and 3 bugs were equally attracted to T. cruzi-infected or uninfected mice. On the contrary, ITS-2 hybrid bugs reached three times more frequently the T. cruzi-infected mouse, compared to the uninfected one, indicating a significant bias toward an infected host. This behavior may explain in part their higher natural infection rate. Further studies should explore the complex and unique interactions among T. cruzi, triatomines vectors, and mammalian hosts, as this may led to new strategies to interfere with transmission cycles and improve Chagas disease control.

Immune defence mechanisms of triatomines against bacteria, viruses, fungi and parasites.

Triatomines are vectors that transmit the protozoan haemoflagellate Trypanosoma cruzi, the causative agent of Chagas disease. The aim of the current review is to provide a synthesis of the immune mechanisms of triatomines against bacteria, viruses, fungi and parasites to provide clues for areas of further research including biological control. Regarding bacteria, the triatomine immune response includes antimicrobial peptides (AMPs) such as defensins, lysozymes, attacins and cecropins, whose sites of synthesis are principally the fat body and haemocytes. These peptides are used against pathogenic bacteria (especially during ecdysis and feeding), and also attack symbiotic bacteria. In relation to viruses, Triatoma virus is the only one known to attack and kill triatomines. Although the immune response to this virus is unknown, we hypothesize that haemocytes, phenoloxidase (PO) and nitric oxide (NO) could be activated. Different fungal species have been described in a few triatomines and some immune components against these pathogens are PO and proPO. In relation to parasites, triatomines respond with AMPs, including PO, NO and lectin. In the case of T. cruzi this may be effective, but Trypanosoma rangeli seems to evade and suppress PO response. Although it is clear that three parasite-killing processes are used by triatomines - phagocytosis, nodule formation and encapsulation - the precise immune mechanisms of triatomines against invading agents, including trypanosomes, are as yet unknown. The signalling processes used in triatomine immune response are IMD, Toll and Jak-STAT. Based on the information compiled, we propose some lines of research that include strategic approaches of biological control.

Ovarian nutritional resources during the reproductive cycle of the hematophagous Dipetalogaster maxima (Hemiptera: Reduviidae): focus on lipid metabolism.

In this study, we have analyzed the changes of the ovarian nutritional resources in Dipetalogaster maxima at representative days of the reproductive cycle: previtellogenesis, vitellogenesis, as well as fasting-induced early and late atresia. As expected, the amounts of ovarian lipids, proteins, and glycogen increased significantly from previtellogenesis to vitellogenesis and then, diminished during atresia. However, lipids and protein stores found at the atretic stages were higher in comparison to those registered at previtellogenesis. Specific lipid staining of ovarian tissue sections evidenced remarkable changes in the shape, size, and distribution of lipid droplets throughout the reproductive cycle. The role of lipophorin (Lp) as a yolk protein precursor was analyzed by co-injecting Lp-OG (where OG is Oregon Green) and Lp-DiI (where DiI is 1,10-dioctadecyl-3,3,30,30-tetramethylindocarbocyanine) to follow the entire particle, demonstrating that both probes colocalized mainly in the yolk bodies of vitellogenic oocytes. Immunofluorescence assays also showed that Lp was associated to yolk bodies, supporting its endocytic pathway during vitellogenesis. The involvement of Lp in lipid delivery to oocytes was investigated in vivo by co-injecting fluorescent probes to follow the fate of the entire particle (Lp-DiI) and its lipid cargo (Lp-Bodipy-FA). Lp-DiI was readily incorporated by vitellogenic oocytes and no lipoprotein uptake was observed in terminal follicles of ovaries at atretic stages. Bodipy-FA was promptly transferred to vitellogenic oocytes and, to a much lesser extent, to previtellogenic follicles and to oocytes of ovarian tissue at atretic stages. Colocalization of Lp-DiI and Lp-Bodipy-FA inside yolk bodies indicated the relevance of Lp in the buildup of lipid and protein oocyte stores during vitellogenesis.

Effects of mammal host diversity and density on the infection level of Trypanosoma cruzi in sylvatic kissing bugs.

Several reports have described host species diversity and identity as the most important factors influencing disease risk, producing either dilution or amplification of the pathogen in a host community. Triatomine vectors, mammals and the protozoan Trypanosoma cruzi (Trypanosomatida: Trypanosomatidae) Chagas are involved in the wild cycle of Chagas disease, in which infection of mammals occurs by contamination of mucous membranes or skin abrasions with insect-infected faeces. We examined the extent to which host diversity and identity determine the infection level observed in vector populations (i.e. disease risk in humans). We recorded infection in triatomine colonies and on the coexisting host mammalian species in semi-arid Chile. Host diversity, and total and infected host species densities are used as predictor variables for disease risk. Disease risk did not correlate with host diversity changes. However, the densities of each infected rodent species were positively associated with disease risk. We suggest that the infected host density surrounding the vector colonies is a relevant variable for disease risk and should be considered to understand disease dynamics. It is crucial to pay attention on the spatial scale of analysis, considering the pattern of vector dispersal, when the relationship between host diversity and disease risk is studied.

Laboratory maintenance and care of Rhodnius prolixus (Hemiptera: Reduviidae) and other Triatominae.

Triatomines (Hemiptera: Reduviidae) are hematophagous insects that transmit Trypanosoma cruzi, the etiological agent for Chagas disease, to humans and other mammals. As medically important vectors, species such as Rhodnius prolixus (Hemiptera: Reduviidae) have long been used as a model organism for physiological studies. Laboratory rearing of triatomines is needed to support vector and parasite research. Many environmental conditions, such as suitable housing containers, light source and duration, temperature, humidity, and density, must be addressed when adapting triatomines from a natural habitat for artificial rearing to create conditions for optimal growth and survival. Food source is also an important factor, as triatomines are considered the obligate blood feeders. Parasites and pathogens present risks not only for triatomines but also for the laboratorians handling them. Equipping an insectary space should apply best practices to ensure community, personnel, and insect health. Various triatomine colonies have been maintained in the Centers for Disease Control and Prevention (CDC) Entomology Branch insectary for over 25 years and have more recently been made available to the research community through the Biodefense and Emerging Infections Research Resources Repository (BEI Resources). The CDC Rhodnius prolixus genome has been characterized and thus represents an opportunity for continued model organism research. In addition to fulfilling requests for live triatomines, inquiries are received for support in establishing new and troubleshooting existing laboratory colonies. To accompany the extensive MR4 manual, Methods in Anopheles Research, procedures for triatomine husbandry have been developed and are shared here to address the aforementioned topics.

Prevalence and Diversity of Trypanosoma cruzi in Triatomine Vectors and Their Blood Meal Sources from South Central Texas, USA.

The prevalence of Trypanosoma cruzi was assessed in 117 triatomine insects from central Texas. The qPCR-based results revealed T. cruzi in 59% of the insects (62 adults and eight nymphs), with overall prevalences of T. cruzi of 0% (0/9), 64% (11/17), 58% (10/17), 73% (30/41), and 57% (19/33) for the Bastrop, Caldwell, Gonzales, Guadalupe, and Hays counties, respectively. Analyses of 18S rRNA fragments confirmed T. cuzi in 81% of these samples. Vectors were identified as Triatoma gerstaeckeri (35% of which 65% were positive for T. cruzi), T. sanguisuga (21%, 43% positive), and Paratriatoma leticularia (0.3%, 100% positive). Food sources were recovered from 29% of the insects. Raccoons were 53% of the blood meals (83% positive for T. cruzi), while the remainder came from a variety of sources, including humans (33% positive), house geckos, Eastern woodrats, plain-bellied water snakes (50% positive), hispid cotton rats (0% positive), chickens (100% positive); Asian forest turtles, bison, and pigs (0% positive). The serendipitous detection of blood meal sources at known minimum distances from the collection of the vector insect enabled us to provide several instances where the insect foraging distance was greater than 400 m. These vector foraging distances are novel information that can assist in our understanding of the landscape dynamics for the spread of the pathogen.