The distribution of triatomine (Hemiptera: Reduviidae) vectors of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) in Illinois and Missouri: historical records and specimen submissions from community science programs.

Triatomine species (kissing bugs) infected with Trypanosoma cruzi are found across the southern United States. The northern limits of Trypanosoma cruzi infected kissing bugs are less understood. The objective of this work was to describe the locations of kissing bugs from Illinois and Missouri based on historical records, submissions to Texas A&M University's (TAMU) Kissing Bug Community Science Program and the Centers for Disease Control and Prevention (CDC), and records from online platforms (iNaturalist, BugGuide, and GBIF) up to and including 2022. A total of 228 records were discovered, including 186 from historical or observation platforms and 42 specimens submitted to TAMU or CDC. Species included Triatoma sanguisuga (221 total records, 9 nymphs) and Triatoma lecticularia (7 records). Notably, nearly all (24/26) records submitted to TAMU were collected indoors. Twelve of the 30 (40%) specimens tested were positive for the presence of T. cruzi, including parasite discrete taxonomic units TcI and TcIV. One triatomine sample had been found in a bed feeding on the submitter; this bug was positive for T. cruzi and had evidence of human blood in its gut. Records suggest a ubiquitous distribution in Missouri and potentially to the northernmost border in Illinois. Further investigations into triatomine distribution and infection status are needed within states assumed to be northern limits in order to create public health and veterinary health messaging and baseline distributional maps from which to measure future range shifts in relation to a changing climate.

Trypanosoma cruzi infection in dogs along the US-Mexico border: R0 changes with vector species composition.

Infection with Trypanosoma cruzi, etiological agent of Chagas disease, is common in US government working dogs along the US-Mexico border. This 3145 km long border comprises four states: Texas (TX), New Mexico (NM), Arizona (AZ) and California (CA) with diverse ecosystems and several triatomine (a.k.a., kissing bug) species, primary vectors of T. cruzi in this region. The kissing bug (Heteroptera: Reduviidae) community ranging from CA to TX includes Triatoma protracta (Uhler), Triatoma recurva (Stål) and Triatoma rubida (Uhler) and becomes dominated by Triatoma gerstaeckeri Stål in TX. Here, we ask if T. cruzi infection dynamics in dogs varies along this border region, potentially reflecting changes in vector species and their vectorial capacity. Using reversible catalytic models of infection, where seropositivity can be lost, we estimated an R0 (Estimate ± S.E.) of 1.192 ± 0.084 for TX and NM. In contrast, seropositivity decayed to zero as dogs aged in AZ and CA. These results suggest that dogs are likely infected by T. cruzi during their training in western TX, with a force of infection large enough for keeping R0 above 1, i.e., the disease endemically established, in TX and NM. In AZ and CA, a lower force of infection, probably associated with different vector species communities and associated vectorial capacity and/or different lineages of T. cruzi, results in dogs decreasing their seropositivity with age.

Collection of triatomines from sylvatic habitats by a Trypanosoma cruzi-infected scent detection dog in Texas, USA.

BACKGROUND: Triatomine insects, vectors of the etiologic agent of Chagas disease (Trypanosoma cruzi), are challenging to locate in sylvatic habitats. Collection techniques used in the United States often rely on methods to intercept seasonally dispersing adults or on community scientists' encounters. Neither method is suited for detecting nest habitats likely to harbor triatomines, which is important for vector surveillance and control. Furthermore, manual inspection of suspected harborages is difficult and unlikely to reveal novel locations and host associations. Similar to a team that used a trained dog to detect sylvatic triatomines in Paraguay, we worked with a trained scent detection dog to detect triatomines in sylvatic locations across Texas. PRINCIPLE METHODOLOGY/FINDINGS: Ziza, a 3-year-old German Shorthaired Pointer previously naturally infected with T. cruzi, was trained to detect triatomines. Over the course of 6 weeks in the fall of 2017, the dog and her handler searched at 17 sites across Texas. The dog detected 60 triatomines at 6 sites; an additional 50 triatomines were contemporaneously collected at 1 of these sites and 2 additional sites without the assistance of the dog. Approximately 0.98 triatomines per hour were found when only humans were conducting searches; when working with the dog, approximately 1.71 triatomines per hour were found. In total, 3 adults and 107 nymphs of four species (Triatoma gerstaeckeri, Triatoma protracta, Triatoma sanguisuga, and Triatoma indictiva) were collected. PCR testing of a subset revealed T. cruzi infection, including DTUs TcI and TcIV, in 27% of nymphs (n = 103) and 66% of adults (n = 3). Bloodmeal analysis of a subset of triatomines (n = 5) revealed feeding on Virginia opossum (Didelphis virginiana), Southern plains woodrat (Neotoma micropus), and eastern cottontail (Sylvilagus floridanus). CONCLUSION/SIGNIFICANCE: A trained scent detection dog enhanced triatomine detections in sylvatic habitats. This approach is effective at detecting nidicolous triatomines. Control of sylvatic sources of triatomines is challenging, but this new knowledge of specific sylvatic habitats and key hosts may reveal opportunities for novel vector control methods to block the transmission of T. cruzi to humans and domestic animals.

Publishing data to support the fight against human vector-borne diseases.

Vector-borne diseases are responsible for more than 17% of human cases of infectious diseases. In most situations, effective control of debilitating and deadly vector-bone diseases (VBDs), such as malaria, dengue, chikungunya, yellow fever, Zika and Chagas requires up-to-date, robust and comprehensive information on the presence, diversity, ecology, bionomics and geographic spread of the organisms that carry and transmit the infectious agents. Huge gaps exist in the information related to these vectors, creating an essential need for campaigns to mobilise and share data. The publication of data papers is an effective tool for overcoming this challenge. These peer-reviewed articles provide scholarly credit for researchers whose vital work of assembling and publishing well-described, properly-formatted datasets often fails to receive appropriate recognition. To address this, GigaScience's sister journal GigaByte partnered with the Global Biodiversity Information Facility (GBIF) to publish a series of data papers, with support from the Special Programme for Research and Training in Tropical Diseases (TDR), hosted by the World Health Organisation (WHO). Here we outline the initial results of this targeted approach to sharing data and describe its importance for controlling VBDs and improving public health.

Characterization of triatomine bloodmeal sources using direct Sanger sequencing and amplicon deep sequencing methods.

Knowledge of host associations of blood-feeding vectors may afford insights into managing disease systems and protecting public health. However, the ability of methods to distinguish bloodmeal sources varies widely. We used two methods-Sanger sequencing and amplicon deep sequencing-to target a 228 bp region of the vertebrate Cytochrome b gene and determine hosts fed upon by triatomines (n = 115) collected primarily in Texas, USA. Direct Sanger sequencing of PCR amplicons was successful for 36 samples (31%). Sanger sequencing revealed 15 distinct host species, which included humans, domestic animals (Canis lupus familiaris, Ovis aries, Gallus gallus, Bos taurus, Felis catus, and Capra hircus), wildlife (Rattus rattus, Incilius nebulifer, Sciurus carolinensis, Sciurus niger, and Odocoileus virginianus), and captive animals (Panthera tigris, Colobus spp., and Chelonoidis carbonaria). Samples sequenced by the Sanger method were also subjected to Illumina MiSeq amplicon deep sequencing. The amplicon deep sequencing results (average of 302,080 usable reads per sample) replicated the host community revealed using Sanger sequencing, and detected additional hosts in five triatomines (13.9%), including two additional blood sources (Procyon lotor and Bassariscus astutus). Up to four bloodmeal sources were detected in a single triatomine (I. nebulifer, Homo sapiens, C. lupus familiaris, and S. carolinensis). Enhanced understanding of vector-host-parasite networks may allow for integrated vector management programs focusing on highly-utilized and highly-infected host species.

American triatomine species occurrences: updates and novelties in the DataTri database.

The causative agent of Chagas disease (Trypanosoma cruzi) is transmitted to mammals, including humans, mainly by insect vectors of the subfamily Triatominae (Hemiptera: Reduviidae). Also known as "kissing bugs", the subfamily currently includes 157 validated species (154 extant and three extinct), in 18 genera and five tribes. Here, we present a subdataset (7852 records) of American triatomine occurrences; an update to the most complete and integrated database available to date at a continental scale. New georeferenced records were obtained from a systematic review of published literature and colleague-provided data. New data correspond to 101 species and 14 genera from 22 American countries between 1935 and 2022. The most important novelties refer to (i) the inclusion of new species, (ii) synonymies and formal transferals of species, and (iii) temporal and geographical species records updates. These data will be a useful contribution to entomological surveillance implicated in Chagas disease.

High incidence of Trypanosoma cruzi infections in dogs directly detected through longitudinal tracking at 10 multi-dog kennels, Texas, USA.

Canine Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, is increasingly recognized as a health concern for dogs in the USA, and infected dogs may signal geographic regions of risk for human disease. Dogs living in multi-dog kennel environments (kennels with more than one dog) where triatomine vectors are endemic may be at high risk for infection. We monitored a cohort of 64 T. cruzi-infected and uninfected dogs across 10 kennels in Texas, USA, to characterize changes in infection status over one year. We used robust diagnostic criteria in which reactivity on multiple independent platforms was required to be considered positive. Among the 30 dogs enrolled as serologically- and/or PCR-positive, all but one dog showed sustained positive T. cruzi diagnostic results over time. Among the 34 dogs enrolled as serologically- and PCR-negative, 10 new T. cruzi infections were recorded over a 12-month period. The resulting incidence rate for dogs initially enrolled as T. cruzi-negative was 30.7 T. cruzi infections per 100 dogs per year. This study highlights the risk of T. cruzi infection to dogs in kennel environments. To protect both dog and human health, there is an urgent need to develop more integrated vector control methods as well as prophylactic and curative antiparasitic treatment options for T. cruzi infection in dogs.

Comparison of the Bacterial Gut Microbiome of North American Triatoma spp. With and Without Trypanosoma cruzi.

Chagas disease, caused by the hemoflagellate protist Trypanosoma cruzi, affects nearly 6 million people worldwide, mainly in Latin America. Hematophagous triatomine insects ("kissing bugs") are the primary vectors of T. cruzi throughout the Americas and feed on a variety of animals, including humans. Control of triatomines is central to the control of T. cruzi infection. Recent advances in mitigation of other insect-borne diseases via the manipulation of insect-associated bacteria as a way to halt or slow disease transmission has opened questions to the applicability of these methods to Chagas disease vectors. Few studies have examined the hindgut microbiome of triatomines found in North America. In the current study, two species of triatomines were collected across Texas, United States, screened for the presence of T. cruzi, and analyzed for the bacterial composition of their hindguts using a 16S rRNA gene-fragment metabarcoding approach. We compared diversity of microbial community profiles across 74 triatomine insects to address the hypothesis that the richness and abundance of bacterial groups differ by T. cruzi infection and strain type, blood meal engorgement status, insect species, sex, and collection location. The gut microbial community of individual triatomines was characterized by low intraindividual taxonomic diversity and high interindividual variation that was weakly predicted by triatomine species, and was not predicted by triatomine sex, collection location, T. cruzi infection status, or blood meal score. However, we did find bacterial groups enriched in T. cruzi-positive individuals, including Enterobacterales, and Petrimonas. Additionally, we detected Salmonella enterica subspecies diarizonae in three triatomine individuals; this species is commonly associated with reptiles and domesticated animals and is a pathogen of humans. These data suggest that Triatoma spp. in Texas have variable patterns of colonized and transient bacteria, and may aid in development of novel means to interfere with transmission of the Chagas disease parasite T. cruzi. Deeper understanding of the effects of parasite infection on diverse insect vector microbiomes may highlight disease transmission risk and facilitate discovery of possible intervention strategies for biological control of this emerging vector-borne disease of global health significance.

Repeated cross-sectional study of Trypanosoma cruzi in shelter dogs in Texas, in the context of Dirofilaria immitis and tick-borne pathogen prevalence.

BACKGROUND: Vector-borne diseases have an adverse impact on health of dogs, and infected dogs can be sentinels for human infection. Infection with Trypanosoma cruzi, an agent of Chagas disease, causes fatal heart disease in dogs across the southern United States but has been neglected from wide-scale prevalence studies. OBJECTIVES: To determine the prevalence of exposure to T. cruzi, Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi, and infection with Dirofilaria immitis among dogs in shelters across Texas and to identify risk factors for T. cruzi seropositivity. ANIMALS: Six hundred and eight dogs. METHODS: This repeated cross-sectional study was performed by collecting blood from ~30 dogs during each of the 3 visits to 7 shelters. We tested serum for antibodies to T. cruzi using 2 tests in series and for antibodies to Ehrlichia spp., Anaplasma spp., and B. burgdorferi and D. immitis antigen using the IDEXX SNAP 4DX Plus point-of-care test. DNA was extracted from blood clots and tested for T. cruzi DNA and strain type via quantitative polymerase chain reactions (qPCR). We used logistic regression to assess risk factors. RESULTS: One hundred ten (18.1%) of 608 dogs were seropositive for T. cruzi. Prevalence of exposure to the other vector-borne agents was: Ehrlichia spp. 3.6%; Anaplasma spp. 6.9%; B. burgdorferi 0.2%; and D. immitis infection 16.0%. Six of 559 (1.1%) dogs were qPCR-positive for T. cruzi. CONCLUSIONS AND CLINICAL IMPORTANCE: T. cruzi seroprevalence was comparable to D. immitis prevalence and higher than seroprevalence of the tick-borne pathogens. T. cruzi is an underrecognized health threat to dogs across Texas and possibly other southern states where triatomine vectors are endemic.

Contributions of citizen scientists to arthropod vector data in the age of digital epidemiology.

Citizen-collected arthropod vectors are useful for epidemiological studies of vector-borne disease, especially since the vectors encountered by the public are the subset of vectors in nature that have a disproportionate impact on health. Programs integrating educational efforts with collecting efforts may be particularly effective for public health initiatives, resulting in an empowered public with knowledge of vector-borne disease prevention. Citizen science programs have been successfully implemented for the collection of unprecedented sample sets of mosquitos, ticks, and triatomines. Cyber infrastructure employed in digital epidemiology-including websites, email, mobile phone apps, and social media platforms-has facilitated vector citizen science initiatives to assess disease risk over vast spatial and temporal scales, advancing research to mitigate vector-borne disease risk.

Parasitic interactions among Trypanosoma cruzi, triatomine vectors, domestic animals, and wildlife in Big Bend National Park along the Texas-Mexico border.

National parks attract millions of visitors each year. Park visitors, employees, and pets are at risk of infection with various zoonotic pathogens, including Trypanosoma cruzi, causative agent of Chagas disease. Big Bend National Park is located along the Texas-Mexico border in a region with endemic triatomine insects- vectors of T. cruzi- yet the degree to which the parasite is transmitted in this region is unknown. We collected triatomines for T. cruzi detection and discrete typing unit (DTU) determination, and conducted blood meal analyses to determine recent hosts. As an index of domestic/peridomestic transmission, we tested residential dogs in the Park for exposure to T. cruzi. From 2015 to 2017, 461 triatomines of three species-Triatoma rubida, Triatoma gerstaeckeri, and Triatoma protracta-were collected in and around the Park. Adult triatomine encounters peaked in June of each year (52.8% of collections). We detected an overall infection prevalence of 23.1% in adult triatomines (n = 320) and 4.2% in nymph triatomines (n = 24). DTU TcI was the only T. cruzi strain detected. Of 89 triatomines subjected to blood meal analyses, vertebrate host DNA was successfully amplified from 42 (47.2%); blood meal sources included humans, domestic animals, and avian and mammalian wildlife species. Tested dogs were considered positive if reactive on at least two independent serologic assays; we found 28.6% seroprevalence in 14 dogs. These findings reveal interactions between infected triatomines, humans, dogs, and wildlife in and around Big Bend National Park, with potential risk of human disease.

A Pilot Radio Telemetry Field Study of Triatomine Vectors (Hemiptera: Reduviidae) of the Chagas Disease Parasite.

We conducted the first pilot radio telemetry study of hematophagous arthropods by placing transmitters on wild-caught triatomine insects ('kissing bugs'), vectors of the Chagas disease parasite. In Texas-a recognized hotspot for triatomine diversity and locally-acquired human and animal Chagas disease-we tagged five female and four male Triatoma gerstaeckeri (Stål) (Hemiptera: Reduviidae), as well as one female and one male Triatoma sanguisuga (Leconte) (Hemiptera: Reduviidae) in three counties from 2015 to 2017. In comparative trials, placement of the transmitter on the dorsal side of the abdomen underneath the hemelytra wings, with the transmitter antenna shortened to 3 cm, yielded the best results. We tracked the movements of the 11 tagged bugs over an average of 4.8 d (range of 1 to 12 d) and detected 18 movement events with an average distance of 3.8 m (range of 1 to 20 m). This pilot study demonstrates the potential utility for using telemetry as a tool for studying fine-scale non-flight movement of triatomines and the discovery of cryptic resting habitats. Future studies using this or similar technologies to study movement and behavior of triatomines could test for site-fidelity of resting habitats and provide novel insight into aspects of vector biology that could be targeted in disease risk reduction efforts.

Clear Resin Casting of Arthropods of Medical Importance for Use in Educational and Outreach Activities.

Arthropod-related morbidity and mortality represent a major threat to human and animal health. An important component of reducing vector-borne diseases and injuries is training the next generation of medical entomologists and educating the public in proper identification of arthropods of medical importance. One challenge of student training and public outreach is achieving a safe mounting technique that allows observation of morphological characteristics, while minimizing damage to specimens that are often difficult to replace. Although resin-embedded specimens are available from commercial retailers, there is a need for a published protocol that allows entomologists to economically create high-quality resin-embedded arthropods for use in teaching and outreach activities. We developed a detailed protocol using readily obtained equipment and supplies for creating resin-embedded arthropods of many species for use in teaching and outreach activities.

Analysis of over 1500 triatomine vectors from across the US, predominantly Texas, for Trypanosoma cruzi infection and discrete typing units.

Across the Americas, triatomine insects harbor diverse strains of Trypanosoma cruzi (T. cruzi), agent of Chagas disease. Geographic patterns of vector infection and parasite strain associations, especially in vectors encountered by the public, may be useful in assessing entomological risk, but are largely unknown across the US. We collected Triatoma spp. from across the US (mainly Texas), in part using a citizen science initiative, and amplified T. cruzi DNA to determine infection prevalence and parasite discrete typing units (DTUs). We found 54.4% infection prevalence in 1510 triatomines of 6 species; prevalence in adult T. gerstaeckeri (63.3%; n=897) and T. lecticularia (66.7%; n=66) was greater than in T. sanguisuga (47.6%; n=315), T. indictiva (47.8% n=67), T. rubida (14.1%; n=64), and T. protracta (10.5%; n=19). The odds of infection in adults were 9.73 times higher than in nymphs (95% CI 4.46-25.83). PCR of the spliced leader intergenic region (SL-IR) and/or the putative lathosterol/episterol oxidase TcSC5D gene revealed exclusively T. cruzi DTUs TcI and TcIV; 5.5% of T. cruzi-positive samples were not successfully typed. T. gerstaeckeri (n=548) were more frequently infected with TcI (53.9%) than TcIV (34.4%), and 11.9% showed mixed TcI/TcIV infections. In contrast, T. sanguisuga (n=135) were more frequently infected with TcIV (79.3%) than TcI (15.6%), and 5.2% showed mixed infections. Relative abundance of parasite DTUs varied spatially, with both TcI and TcIV co-circulating in vectors in central Texas, while TcIV predominated in northern Texas. Given prior findings implicating TcI in human disease and TcI and TcIV in animal disease in the US, knowledge of spatial distribution of T. cruzi infection and DTUs in vectors is important to understanding public and veterinary health risk of T. cruzi infection.

Bionomics and Spatial Distribution of Triatomine Vectors of Trypanosoma cruzi in Texas and Other Southern States, USA.

Defining spatial and temporal occurrences of triatomine vectors of Trypanosoma cruzi, the agent of Chagas disease, in the US is critical for public health protection. Through a citizen science program and field collections from 2012 to 2016, we collected 3,215 triatomines, mainly from Texas. Using morphological and molecular approaches, we identified seven Triatoma species and report sex, length, and blood engorgement status. Many citizen-collected triatomines (92.9%) were encountered indoors, in peridomestic settings, or in dog kennels and represent spillover transmission risk of T. cruzi to humans and domestic animals. The most commonly collected species were Triatoma gerstaeckeri and Triatoma sanguisuga. Adult T. gerstaeckeri were collected from May to September, peaking from June to July, whereas adult T. sanguisuga were active later, from June to October, peaking from July to September. Based on cross correlation analyses, peaks of captures varied by species and across years. Point pattern analyses revealed unique occurrences of T. sanguisuga in north and east Texas, T. gerstaeckeri in south and west Texas, Triatoma indictiva and Triatoma lecticularia in central Texas, and Triatoma rubida in west Texas. These relatively unique spatial occurrences suggest associations with different suitable habitats and serve as a basis for future models evaluating the ecological niches of different vector species. Understanding the temporal and spatial heterogeneity of triatomines in the southern United States will improve targeted interventions of vector control and will guide public outreach and education to reduce human and animal contact with vectors and reduce the risk of exposure to T. cruzi.

Trypanosoma cruzi (Agent of Chagas Disease) in Sympatric Human and Dog Populations in "Colonias" of the Lower Rio Grande Valley of Texas.

AbstractThe zoonotic, vector-borne parasite Trypanosoma cruzi causes Chagas disease throughout the Americas, but human and veterinary health burdens in the United States are unknown. We conducted a cross-sectional prevalence study in indigent, medically underserved human and cohabiting canine populations of seven south Texas border communities, known as colonias. Defining positivity as those samples that were positive on two or more independent tests, we found 1.3% seroprevalence in 233 humans, including one child born in the United States with only short-duration travel to Mexico. Additionally, a single child with no travel outside south Texas was positive on only a single test. Among 209 dogs, seroprevalence was 19.6%, but adjusted to 31.6% when including those dogs positive on only one test and extrapolating potential false negatives. Parasite DNA was detected in five dogs, indicating potential parasitemia. Seropositive dogs lived in all sampled colonias with no difference in odds of positivity across age, sex, or breed. Colonia residents collected two adult Triatoma gerstaeckeri and one nymph triatomine from around their homes; one of three bugs was infected with T. cruzi, and blood meal hosts were molecularly determined to include dog, human, and raccoon. Dogs and the infected vector all harbored T. cruzi discrete typing unit I, which has previously been implicated in human disease in the United States. Colonias harbor active T. cruzi transmission cycles and should be a priority in outreach and vector control initiatives.

Epidemiology and Molecular Typing of Trypanosoma cruzi in Naturally-Infected Hound Dogs and Associated Triatomine Vectors in Texas, USA.

BACKGROUND: Trypanosoma cruzi is the etiologic agent of Chagas disease throughout the Americas. Few population-level studies have examined the epidemiology of canine infection and strain types of T. cruzi that infect canines in the USA. We conducted a cross-sectional study of T. cruzi infection in working hound dogs in south central Texas, including analysis of triatomine vectors collected within kennel environments. METHODOLOGY/PRINCIPLE FINDINGS: Paired IFA and Chagas Stat-Pak serological testing showed an overall seroprevalence of 57.6% (n = 85), with significant variation across kennels. Dog age had a marginally significant effect on seropositivity, with one year of age increase associated with a 19.6% increase in odds of being seropositive (odds ratio 95% CI 0.996-1.435; p = 0.055). PCR analyses of blood revealed 17.4% of dogs harbored parasite DNA in their blood, including both seronegative and seropositive dogs. Molecular screening of organs from opportunistically sampled seropositive dogs revealed parasite DNA in heart, uterus, and mammary tissues. Strain-typing showed parasite discrete typing units (DTU) TcI and TcIV present in dog samples, including a co-occurrence of both DTUs in two individual dogs. Bloodmeal analysis of Triatoma gerstaeckeri and Triatoma sanguisuga insects collected from the kennels revealed exclusively dog DNA. Vector infection with T. cruzi was 80.6% (n = 36), in which T. gerstaeckeri disproportionately harbored TcI (p = 0.045) and T. sanguisuga disproportionately harbored TcIV (p = 0.029). Tracing infection status across dog litters showed some seropositive offspring of seronegative dams, suggesting infection of pups from local triatomine vectors rather than congenital transmission. CONCLUSIONS/SIGNIFICANCE: Canine kennels are high-risk environments for T. cruzi transmission, in which dogs likely serve as the predominant parasite reservoir. Disease and death of working dogs from Chagas disease is associated with unmeasured yet undoubtedly significant financial consequences because working dogs are highly trained and highly valued.

High Trypanosoma cruzi infection prevalence associated with minimal cardiac pathology among wild carnivores in central Texas.

Infection with the zoonotic vector-borne protozoal parasite Trypanosoma cruzi causes Chagas disease in humans and dogs throughout the Americas. Despite the recognized importance of various wildlife species for perpetuating Trypanosoma cruzi in nature, relatively little is known about the development of cardiac disease in infected wildlife. Using a cross-sectional study design, we collected cardiac tissue and blood from hunter-donated wildlife carcasses- including raccoon (Procyon lotor), coyote (Canis latrans), gray fox (Urocyon cinereoargenteus), and bobcat (Lynx rufus) - from central Texas, a region with established populations of infected triatomine vectors and increasing diagnoses of Chagas disease in domestic dogs. Based on PCR analysis, we found that 2 bobcats (14.3%), 12 coyotes (14.3%), 8 foxes (13.8%), and 49 raccoons (70.0%) were positive for T. cruzi in at least one sample (right ventricle, apex, and/or blood clot). Although a histologic survey of right ventricles showed that 21.1% of 19 PCR-positive hearts were characterized by mild lymphoplasmocytic infiltration, no other lesions and no amastigotes were observed in any histologic section. DNA sequencing of the TcSC5D gene revealed that raccoons were infected with T. cruzi strain TcIV, and a single racoon harbored a TcI/TcIV mixed infection. Relative to other wildlife species tested here, our data suggest that raccoons may be important reservoirs of TcIV in Texas and a source of infection for indigenous triatomine bugs. The overall high level of infection in this wildlife community likely reflects high levels of vector contact, including ingestion of bugs. Although the relationship between the sylvatic cycle of T. cruzi transmission and human disease risk in the United States has yet to be defined, our data suggest that hunters and wildlife professionals should take precautions to avoid direct contact with potentially infected wildlife tissues.

Survey of Feral Swine ( Sus scrofa ) Infection with the Agent of Chagas Disease ( Trypanosoma cruzi ) in Texas, 2013-14.

: Feral swine ( Sus scrofa ) are an invasive species and reservoir of numerous zoonotic pathogens in the US, and Texas leads the nation in the estimated population size of feral hogs. Texas also harbors enzootic transmission cycles of the protozoan parasite Trypanosoma cruzi , agent of Chagas disease. Given previous evidence that swine can serve as reservoirs of T. cruzi in Latin America and new evidence of triatomines (kissing bugs) feeding on swine in Texas, we measured the prevalence of T. cruzi infection in feral swine in Texas. From 2013 to 2014, we sampled blood and/or cardiac tissue from 78 feral swine across 14 Texas counties (seven with and seven without prior documentation of kissing bug occurrence) and used PCR and histopathology to detect T. cruzi infection. We determined an overall infection prevalence of 6% (3 of 54) based on PCR evaluation of cardiac tissue, and no blood samples were positive (n=72). All three positive pigs were from counties where kissing bugs are documented. No T. cruzi amastigotes were noted on histopathology (n=54). Sarcocysts were observed in 10 (18%) of the samples, five of which also had mild focal areas of degeneration and inflammatory cell infiltration. Eco-epidemiologic investigations can provide an assessment of contributions of feral hogs to maintenance of T. cruzi across a landscape to help protect human and animal health.

Chagas disease in a Texan horse with neurologic deficits.

A 10-year-old Quarter Horse gelding presented to the Texas A&M University Veterinary Teaching Hospital with a six month-history of ataxia and lameness in the hind limbs. The horse was treated presumptively for equine protozoal myeloencephalitis (EPM) based on clinical signs but was ultimately euthanized after its condition worsened. Gross lesions were limited to a small area of reddening in the gray matter of the thoracic spinal cord. Histologically, trypanosome amastigotes morphologically similar to Trypanosoma cruzi, the agent of Chagas disease in humans and dogs, were sporadically detected within segments of the thoracic spinal cord surrounded by mild lymphoplasmacytic inflammation. Ancillary testing for Sarcocystis neurona, Neospora spp., Toxoplasma gondii and Leishmania spp. was negative. Conventional and real time polymerase chain reaction (PCR) of affected paraffin embedded spinal cord were positive for T. cruzi, and sequencing of the amplified T. cruzi satellite DNA PCR fragment from the horse was homologous with various clones of T. cruzi in GenBank. While canine Chagas disease cases have been widely reported in southern Texas, this is the first report of clinical T. cruzi infection in an equid with demonstrable amastigotes in the spinal cord. In contrast to previous instances of Chagas disease in the central nervous system (CNS) of dogs and humans, no inflammation or T. cruzi amastigotes were detected in the heart of the horse. Based on clinical signs, there is a potential for misdiagnosis of Chagas disease with other infectious diseases that affect the equine CNS. T. cruzi should be considered as a differential diagnosis in horses with neurologic clinical signs and histologic evidence of meningomyelitis that originate in areas where Chagas disease is present. The prevalence of T. cruzi in horses and the role of equids in the parasite life cycle require further study.