Rachel Curtis-Robles

Shelter dogs as sentinels for Trypanosoma cruzi transmission across Texas.

Chagas disease, an infection with the parasite Trypanosoma cruzi, is increasingly diagnosed among humans in the southern United States. We assessed exposure of shelter dogs in Texas to T. cruzi; seroprevalence across diverse ecoregions was 8.8%. Canine serosurveillance is a useful tool for public health risk assessment.

Combining Public Health Education and Disease Ecology Research: Using Citizen Science to Assess Chagas Disease Entomological Risk in Texas.

Background Chagas disease is a zoonotic parasitic disease well-documented throughout the Americas and transmitted primarily by triatomine ‘kissing bug’ vectors. In acknowledgment of the successful history of vector control programs based on community participation across Latin America, we used a citizen science approach to gain novel insight into the geographic distribution, seasonal activity, and Trypanosoma cruzi infection prevalence of kissing bugs in Texas while empowering the public with information about Chagas disease. Methodology/Principal Findings We accepted submissions of kissing bugs encountered by the public in Texas and other states from 2013–2014 while providing educational literature about Chagas disease. In the laboratory, kissing bugs were identified to species, dissected, and tested for T. cruzi infection. A total of 1,980 triatomines were submitted to the program comprised of at least seven species, of which T. gerstaeckeri and T. sanguisuga were the most abundant (85.7% of submissions). Triatomines were most commonly collected from dog kennels and outdoor patios; Overall, 10.5% of triatomines were collected from inside the home. Triatomines were submitted from across Texas, including many counties which were not previously known to harbor kissing bugs. Kissing bugs were captured primarily throughout April-October, and peak activity occurred in June-July. Emails to our dedicated account regarding kissing bugs were more frequent in the summer months (June-August) than the rest of the year. We detected T. cruzi in 63.3% of tested bugs. Conclusions/Significance Citizen science is an efficient approach for generating data on the distribution, phenology, and infection prevalence of kissing bugs—vectors of the Chagas disease parasite—while educating the public and medical community.

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.

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.

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.

Trypanosomacruzi (Agent of Chagas Disease) in Sympatric Human and Dog Populations in “Colonias” of the Lower Rio Grande Valley of Texas

The 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.

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

Abstract. 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.

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

Abstract: 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.

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

Abstract 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. InTexas—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.