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Abstract Trypanosoma cruzi infection in dogs can cause heart failure and sudden death with few treatment options available. A litter of 4 dogs living in a T cruzi endemic area were randomized to prophylaxis and nonprophylaxis groups as part of a study evaluating a modified benznidazole dosing regimen administered twice weekly to prevent T cruzi infection during a vector transmission season. The 2 dogs that received prophylaxis remained healthy without T cruzi infection or cardiac disease for >2 years. One dog that did not receive prophylaxis died unexpectedly with acute T cruzi–induced pancarditis, and the second dog tested positive for T cruzi and developed complex arrhythmias with markedly increased cardiac troponin I and improved with a higher benznidazole treatment dose. Although the small sample size precludes definitive conclusions, we describe the potential clinical benefit of prophylactic and early treatment with modified benznidazole dosing regimens for dogs with T cruzi infection.

Dirofilaria immitis and Dirofilaria striata (Spirurida: Onchocercidae) are epidemiologically important filarial nematodes detected in wild carnivores sympatric to domestic animals and humans. In this study we surveyed for Dirofilaria species among previous studies archived blood samples (n = 202) of wild carnivores sourced across Texas between the years of 2014–2016 and 2020 to 2023. In total, 117 coyotes (Canis latrans), 67 raccoons (Procyon lotor), 12 gray foxes (Urocyon cinereoargenteus), five bobcats (Lynx rufus), and one striped skunk (Mephitis mephitis) were tested through the amplification of the partial cytochrome oxidase c subunit 1 (COI) gene followed by sequencing. Dirofilaria immitis was detected in 11.39% (95% CI = 7.71–16.51) of the samples (21 coyotes and two raccoons), while D. striata was detected in a single bobcat. Dirofilaria immitis sequences had 99.85%–100% (99.92% ± 0.08) similarity with other D. immitis sequences in GenBank. The sequence of D. striata from the bobcat was 100% similar to the single COI sequence available in GenBank. Data from this study reinforce the role of coyotes as a wild reservoir for D. immitis and suggest that raccoons may also play a role in the epidemiology of this parasite. This study additionally provides molecular data on D. striata, an understudied filarioid of felids.

Abstract Domestic dogs are susceptible to numerous vector-borne pathogens that are of significant importance for their health. In addition to being of veterinary importance, many of these pathogens are zoonotic and thus may pose a risk to human health. In the USA, owned dogs are commonly screened for exposure to or infection with several canine vector-borne pathogens. Although the screening data are widely available to show areas where infections are being diagnosed, testing of owned dogs is expected to underestimate the actual prevalence in dogs that have no access to veterinary care. The goal of this study was to measure the association between the widely available data from a perceived low-risk population with temporally and spatially collected data from shelter-housed dog populations. These data were then used to extrapolate the prevalence in dogs that generally lack veterinary care. The focus pathogens included Dirofilaria immitis, Ehrlichia spp., Anaplasma spp., and Borrelia burgdorferi. There was a linear association between the prevalence of selected vector-borne pathogens in shelter-housed and owned dog populations and, generally, the data suggested that prevalence of heartworm (D. immitis) infection and seroprevalence of Ehrlichia spp. and B. burgdorferi are higher in shelter-housed dogs, regardless of their location, compared with the owned population. The seroprevalence of Anaplasma spp. was predicted to be higher in areas that have very low to low seroprevalence, but unexpectedly, in areas of higher seroprevalence within the owned population, the seroprevalence was expected to be lower in the shelter-housed dog population. If shelters and veterinarians make decisions to not screen dogs based on the known seroprevalence of the owned group, they are likely underestimating the risk of exposure. This is especially true for heartworm. With this new estimate of the seroprevalence in shelter-housed dogs throughout the USA, shelters and veterinarians can make evidence-based informed decisions on whether testing and screening for these pathogens is appropriate for their local dog population. This work represents an important step in understanding the relationships in the seroprevalences of vector-borne pathogens between shelter-housed and owned dogs, and provides valuable data on the risk of vector-borne diseases in dogs. Graphical abstract

Abstract Molecular methods to understand host feeding patterns of arthropod vectors are critical to assess exposure risk to vector‐borne disease and unveil complex ecological interactions. We build on our prior work discovering the utility of PCR‐Sanger sequencing blood meal analysis that work well for soft ticks (Acari: Argasidae), unlike for hard ticks (Acari: Ixodidae), thanks to their unique physiology that retains prior blood meals for years. Here, we apply blood meal metabarcoding using amplicon deep sequencing to identify multiple host species in individual Ornithodoros turicata soft ticks collected from two natural areas in Texas, United States. Of 788 collected O. turicata, 394 were evaluated for blood meal source via metabarcoding, revealing 27 different vertebrate hosts (17 mammals, five birds, one reptile, and four amphibians) fed upon by 274 soft ticks. Information on multiple hosts was derived from 167 individual O. turicata (61%). Metabarcoding revealed mixed vertebrate blood meals in O. turicata while same specimens yielded only one vertebrate species using Sanger sequencing. These data reveal wide host range of O. turicata and demonstrate the value of blood meal metabarcoding for understanding the ecology for known and potential tick‐borne pathogens circulating among humans, domestic animals, and wildlife such as relapsing fever caused by Borrelia turicatae. Our results also document evidence of prior feeding on wild pig from an off‐host soft tick for the first time in North America; a critical observation in the context of enzootic transmission of African swine fever virus if it were introduced to the US. This research enhances our understanding of vector‐host associations and offers a promising perspective for biodiversity monitoring and disease control strategies.

Abstract Background Chagas disease remains a persistent vector-borne neglected tropical disease throughout the Americas and threatens both human and animal health. Diverse control methods have been used to target triatomine vector populations, with household insecticides being the most common. As an alternative to environmental sprays, host-targeted systemic insecticides (or endectocides) allow for application of chemicals to vertebrate hosts, resulting in toxic blood meals for arthropods (xenointoxication). In this study, we evaluated three systemic insecticide products for their ability to kill triatomines. Methods Chickens were fed the insecticides orally, following which triatomines were allowed to feed on the treated chickens. The insecticide products tested included: Safe-Guard® Aquasol (fenbendazole), Ivomec® Pour-On (ivermectin) and Bravecto® (fluralaner). Triatoma gerstaeckeri nymphs were allowed to feed on insecticide-live birds at 0, 3, 7, 14, 28 and 56 days post-treatment. The survival and feeding status of the T. gerstaeckeri insects were recorded and analyzed using Kaplan–Meier curves and logistic regression. Results Feeding on fluralaner-treated chickens resulted 50–100% mortality in T. gerstaeckeri over the first 14 days post-treatment but not later; in contrast, all insects that fed on fenbendazole- and ivermectin-treated chickens survived. Liquid chromatography tandem mass spectrometry (LC-QQQ) analysis, used to detect the concentration of fluralaner and fenbendazole in chicken plasma, revealed the presence of fluralaner in plasma at 3, 7, and 14 days post-treatment but not later, with the highest concentrations found at 3 and 7 days post-treatment. However, fenbendazole concentration was below the limit of detection at all time points. Conclusions Xenointoxication using fluralaner in poultry is a potential new tool for integrated vector control to reduce risk of Chagas disease. Graphical Abstract

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.

Trypanosoma cruzi , the protozoan parasite and cause of Chagas disease, is widely distributed in many vertebrate and triatomine species throughout North, Central, and South America. Variations in housing quality largely determines human infection risk in the Americas. However, the southern U.S. contains widespread, infected triatomine vectors and captive species and domesticated animals with active T. cruzi infection or at high risk of becoming infected and developing Chagas disease. There is a critical need for better detection and intervention strategies, principally focused on human infection throughout the Americas, but mainly in the U.S., for high-value dogs employed in government and other work. In addition to this economic impact, the concentration of largely unavoidable T. cruzi infections in U.S. dogs provides an incomparable opportunity to answer questions related to T. cruzi infection and Chagas disease that are impossible or unethical to address in humans. As the course of T. cruzi infection and Chagas disease, the immune response to infection, and the response to therapeutics are highly similar across the range of mammalian host species, information obtained from studies in other species can directly inform researchers on how to best detect, manage, and treat T. cruzi infection and Chagas disease in humans.

After identifying a captive herd of white-tailed deer in central Texas with >94% seroprevalence with SARS-CoV-2 neutralizing antibodies in September 2021, we worked retrospectively through archived serum samples of 21 deer and detected seroconversion of all animals between December 2020 and January 2021. We then collected prospective samples to conclude that the duration of persistence of neutralizing antibodies is at least 13 months for 19 (90.5%) of the animals, with two animals converting to seronegative after six and eight months. Antibody titres generally waned over this time frame, but three deer had a temporary 4- to 8-fold increases in plaque reduction neutralization test titres over a month after seroconversion; anamnestic response cannot be ruled out.

Abstract Background The canine heartworm Dirofilaria immitis, a filarioid nematode of dogs and other carnivores, is widespread in the USA and the world. Over 20 different mosquito species serve as intermediate hosts of D. immitis, but their contribution to transmission varies according to factors like host feeding patterns, geographic locations and climatic conditions. The yellow fever mosquito, Aedes aegypti, is a competent vector of D. immitis but is often dismissed as a vector of veterinary relevance given its anthropophilic feeding behavior. We evaluated the prevalence of D. immitis in pet dogs along the USA-Mexico border and assessed whether Ae. aegypti in the area are naturally infected with heartworm and are potentially acting as a vector. Methods A total of 200 whole blood samples collected from pet dogs in the Lower Rio Grande Valley in south Texas from 2016 to 2019 were included in this study. Canine serum samples for D. immitis were tested using the DiroCHEK® Canine Heartworm Antigen Test Kit pre- and post-immune complex dissociations (ICD) and blood samples were tested using high-resolution melt (HRM) quantitative PCR (qPCR) and a probe-based qPCR. Additionally, mosquito specimens were collected and identified, and Ae. aegypti heads, abdomens and pools were tested using conventional PCR (cPCR) and HRM qPCR. Results Overall, heartworm prevalence in dogs aged > 6 months was 40.8% (64/157) when the results from all testing modalities were considered. Heartworm antigen was detected in 33.5% and 40.7% of the dogs using DiroCHEK® pre- and post-ICD, respectively. By molecular screening, 20.1% of dogs tested positive with probe-based qPCR, while only one tested positive with HRM qPCR. Of the Ae. aegypti abdomens from blood-fed Ae. aeygpti tested, 20 (21.7%) from mosquitoes that fed on dogs and four (7%) from those that fed on humans tested positive for heartworm. Among Ae. aegypti heads from blood-fed Ae. aeygpti, two (1.1%) were positive based on cPCR and four (2.5%) were positive based on HRM qPCR. No D. immitis DNA was detected in the 208 pools of whole bodies (358 individuals) of Ae. aegypti gravid females. Conclusions Our study highlights a high prevalence of heartworm in dogs in south Texas and provides evidence that Ae. aegypti could be contributing to heartworm transmission in canine populations in this region. Graphical Abstract

Abstract 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.

We tested 294 domestic pet dogs in Mexico for neutralizing antibodies for mosquito-borne flaviviruses. We found high (42.6%) exposure to West Nile virus in Reynosa (northern Mexico) and low (1.2%) exposure in Tuxtla Gutierrez (southern Mexico) but very limited exposure to Aedes-borne flaviviruses. Domestic dogs may be useful sentinels for West Nile virus.

Abstract Chagas cardiomyopathy, caused by the protozoal parasite Trypanosoma cruzi, is characterized by arrhythmias, myocardial damage, heart failure, and sudden death. We describe 2 dogs with severe, symptomatic Chagas cardiomyopathy characterized by myocardial dysfunction and electrocardiographic abnormalities that were managed with a combination of cardiac medications and antiparasitic treatment with itraconazole and amiodarone. Both dogs died suddenly within 6 months of diagnosis. These cases highlight the need for early detection of Chagas disease in dogs and continued research to develop effective antiparasitic treatment protocols.

Texas is a geographically large state with large human and livestock populations, many farms, a long coastal region, and extreme fluctuations in weather. During the last 15 years, the state of Texas has frequently suffered disasters or catastrophes causing extensive morbidity and economic loss. These disasters often have complicated consequences requiring multi-faceted responses. Recently, an interdisciplinary network of professionals from multiple academic institutions has emerged to collaborate in protecting Texas and the USA using a One Health approach. These experts are training the next generation of scientists in biopreparedness; increasing understanding of pathogens that cause repetitive harm; developing new therapeutics and vaccines against them; and developing novel surveillance approaches so that emerging pathogens will be detected early and thwarted before they can cause disastrous human and economic losses. These academic One Health partnerships strengthen our ability to protect human and animal health against future catastrophes that may impact the diverse ecoregions of Texas and the world.

Animal movements, especially avian migration, can be a mechanism for the large-scale dispersal and geographic range expansion of parasites. The host-parasite relationships among birds during migration have yet to be fully explored. We characterized the haemosporidian parasite lineages in passerines during spring migration on the Texas coast of the Gulf of Mexico, and identified associations among wintering origin (US, Central America, South America) and foraging height (canopy, understory, ground) and infection status. We examined 743 samples representing 52 species of 10 families over six years, 2014–2019. We used PCR and DNA sequencing of the haemosporidian cytB gene from avian blood samples to determine infection status with the genera Plasmodium and Haemoproteus and characterize the lineages of blood parasites. We found an overall haemosporidian infection prevalence of 48.4% among neotropical migrant and Texas wintering birds. Among families, Icterids had the highest prevalence (75%, 24 individuals, 4 species sampled) whereas Parulids had the lowest prevalence (38.4%, 177 individuals, 18 species sampled). Among infected birds, Plasmodium spp. infections were more common than Haemoproteus spp. infections in species that winter in Central America compared to those that winter in the US or South America. Similarly, among infected birds, Plasmodium spp. infections were more common than Haemoproteus spp. infections in species that forage on the ground or in the understory compared to those that forage in the canopy. Infected birds harbored 65 different haemosporidian lineages (71% Plasmodium; 29% Haemoproteus) of which 17 lineages have never previously been reported and six lineages were documented for the first time in North America, having been previously detected only in Central or South America. These data are consistent with the premise that intercontinental parasite dispersal may be facilitated by passerine birds. Future studies focused on surveillance, the probability of establishment of parasite lineages, and the use of individual bird tracking methods to understand infection dispersion over time will allow a more comprehensive understanding of changing avian host-haemosporidian relationships.

Abstract Background The protozoal parasite Trypanosoma cruzi causes myocarditis in dogs. Objectives To describe the cardiac diagnostic test results and outcomes of dogs naturally infected with T. cruzi. Animals Forty‐four client‐owned dogs. Methods Medical records were retrospectively reviewed to identify dogs with an indirect fluorescent antibody test result for T. cruzi ≥1 : 80. Data collected included signalment, cardiac diagnostic test results (ECG, echocardiography, cardiac troponin I) and outcome. Outcomes were categorized as alive, dead (cardiac or noncardiac) or lost to follow up. Results ECG abnormalities were present in 41 dogs with ventricular arrhythmias (n = 28) and atrioventricular block (AVB) (n = 15) most commonly identified. Echocardiographic chamber enlargement was present in 28 dogs and most often included the right ventricle (RV) (n = 15) and left atrium (n = 12). Troponin was ≥2 times the reference range in 20/36 (56%) dogs. In univariate analysis using nonparametric Kaplan‐Meier, ventricular arrhythmias with a modified Lown score ≥2 (P = .02), presence of AVB (P = .04), and RV enlargement (P = .006) were associated with decreased survival times. Right ventricular enlargement (HR 3.6; 95% confidence interval [CI] 1.4‐9.3; P = .007) and higher body weight at presentation (HR 1.0; 95% CI 1.0‐1.1; P = .04) were associated with decreased time to death in the final explanatory multivariable model. Conclusions and Clinical Importance Cardiac abnormalities were common and variable, and RV enlargement was associated with shorter survival time. A diagnostic evaluation that includes screening for arrhythmias, echocardiography, and cTnI can provide useful information related to the characterization of heart disease in dogs seropositive for T. cruzi.

Trypanosoma cruzi infection causes inflammation and fibrosis, resulting in cardiac damage in dogs. The objectives of this study were to describe cardiac magnetic resonance imaging (CMR) in naturally infected dogs with chronic Chagas disease and the frequency of abnormalities for CMR and cardiac diagnostic tests. Ten asymptomatic, client-owned dogs seropositive for T. cruzi were prospectively enrolled in an observational study evaluating echocardiography, ECG (standard and ambulatory), cardiac troponin I (cTnI), and CMR. Standard ECG measurements (3/10) and cTnI concentration (1/10) outside the reference range were uncommon. Ambulatory ECG abnormalities were documented more frequently (6/10 dogs) than with standard ECG and included ventricular arrhythmias (4), supraventricular premature beats (3), second-degree atrioventricular block (2), and sinus arrest (1). Echocardiographic abnormalities were documented in 6/10 dogs including mildly increased left ventricular internal dimension in diastole (1) and decreased right ventricular (RV) systolic function based on reductions in tricuspid annular plane systolic excursion (3) and RV S’ (4). Abnormalities were detected with CMR in 7/10 dogs including delayed myocardial enhancement in 5 of which 2 also had increased extracellular volume, abnormal wall motion in 5, and loss of apical compact myocardium in 1. In conclusion, CMR abnormalities were common, and the results of this study suggest CMR can provide useful information in dogs with T. cruzi infection and may support naturally infected dogs for future clinical investigation as an animal model for Chagas disease.

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.

ABSTRACT Free-ranging white-tailed deer (Odocoileus virginianus) across the United States are increasingly recognized for infection and transmission of SARS-CoV-2. Through a cross-sectional study of 80 deer at three captive cervid facilities in central and southern Texas, we provide evidence of 34 of 36 (94.4%) white-tailed deer at a single captive cervid facility seropositive for SARS-CoV-2 by neutralization assay (PRNT90), with endpoint titers as high as 1,280. In contrast, all tested white-tailed deer and axis deer (Axis axis) at two other captive cervid facilities were seronegative, and SARS-CoV-2 RNA was not detected in respiratory swabs from deer at any of the three facilities. These data support transmission among captive deer that cannot be explained by human contact for each infected animal, as only a subset of the seropositive does had direct human contact. The facility seroprevalence was more than double of that reported from wild deer, suggesting that the confined environment may facilitate transmission. Further exploration of captive cervids and other managed animals for their role in the epizootiology of SARS-CoV-2 is critical for understanding impacts on animal health and the potential for spillback transmission to humans or other animal taxa. IMPORTANCE As SARS-CoV-2 vaccine coverage of the human population increases and variants of concern continue to emerge, identification of the epidemiologic importance of animal virus reservoirs is critical. We found that nearly all (94.4%) of the captive white-tailed deer at a cervid facility in central Texas had neutralizing antibodies for SARS-CoV-2. This seroprevalence is over double than that which has been reported from free-ranging deer from other regions of the United States. Horizontal transmission among deer may be facilitated in confinement. Tracking new infections among wild and confined deer is critical for understanding the importance of animal reservoirs for both veterinary and human health.

Trypanosoma cruzi, a zoonotic protozoan parasite, infects a wide range of mammals. The southern United States has endemic sylvatic transmission cycles maintained by several species of wildlife and domestic dogs. We hypothesized that urban-dwelling opossums (Didelphis virginiana) in South Texas are infected with T. cruzi, and that tissue pathology would be associated with infection. In 2017, we collected blood, heart tissue and anal gland secretions from 100 wild opossums across three seasons that were trapped by animal control in South Texas. In addition, anal gland tissue and intercostal muscle were collected from 43 of the 100 opossums for which time allowed the extra tissue collection. All blood, tissue, and secretion samples were screened for T. cruzi DNA using qPCR with confirmation of positive status achieved through one or more additional PCR assays, including a qPCR to determine the parasite discrete typing unit (DTU). T. cruzi DNA was detected in at least one tissue of 15% of the opossums sampled: blood clot (9%), heart tissue (10%), anal gland secretions (12%), intercostal muscle (16.3%), and anal gland tissue (11.6%). Infection was detected in two or more different tissue types in nine of the opossums. The 35 tissues for which parasite DTU was determined were exclusively ‘Tcl’- a DTU previously associated with locally-acquired human disease in the United States. T. cruzi-positive opossums were nearly 14 times more likely to exhibit significant heart lesions on histopathology (lympoplasmacytic inflammation±fibrosis) when compared to negative opossums (OR = 13.56, CI = 1.23–751.28, p-value = 0.03). Three triatomines were opportunistically collected from the study site, of which two were infected (66.7%), and bloodmeal analysis revealed canine, opossum, and human bloodmeals. Given the presence of parasite in opossum blood, unique potential for shedding of parasite in anal glad secretions, and evidence of vectors feeding on opossums, it is likely that opossums serve as wild reservoirs around urban dwellings in South Texas. Keywords: Chagas disease, Texas, Kissing bugs, Triatomine, Urban wildlife

Abstract Background Chagas disease is increasingly recognized in the southern U.S., where triatomine vectors transmit Trypanosoma cruzi among wildlife and domestic dogs with occasional vector spillover to humans. As in humans, clinical outcome in dogs is variable, ranging from acute death to asymptomatic infections or chronic heart disease. In order to characterize cardiac manifestations of T. cruzi infections, we tracked a cohort of naturally-infected dogs and a matched cohort of uninfected dogs. We hypothesized that selected measures of cardiac disease (abnormal rate, abnormal rhythm, and elevated cardiac troponin I (cTnI; a biomarker of cardiac injury)) would occur more commonly in infected than uninfected dogs matched by age, breed, sex and location. In addition to the clearly positive and negative dogs, we specifically tracked dogs with discordant test results across three independent serological assays to gather clinical data that might elucidate the infection status of these animals and inform the utility of the different testing approaches. Results We placed an ambulatory ECG monitor (Holter) on 48 government working dogs and analyzed 39 successful recordings that met length and quality criteria from 17 T. cruzi-infected, 18 uninfected dogs and 4 dogs with discordant results. Overall, 76.5% of positive, 100.0% of discordant, and 11.1% of negative dogs showed > 1 ECG abnormality (p

In Latin America, Mexico is the country with the second highest annual estimated number of Chagas disease cases, caused by Trypanosoma cruzi, due to vector-borne transmission. The state of Oaxaca is the location of the first documented human cases of Chagas disease in Mexico and contained the highest T. cruzi seropositive rate (3.5%) from blood donors. Here, entomological surveys, from 2017 to 2019, were conducted to collect triatomines in 124 villages of 60 municipalities. Four principal domestic Triatoma spp. (Hemiptera: Triatominae), Triatoma phyllosoma, T. barberi, T. mazzotti, and T. dimidiata, of Oaxaca, Mexico were identified by morphology and molecular analysis of the barcode region of the cytochrome oxidase 1 (cox1 or COI or CO1) gene. A total of 41 out of 83 T. phyllosoma specimens examined by microscopy were positive for T. cruzi (49%), 49 out of 171 for T. barberi (28%), 31 out of 177 for T. mazzotti (17%), and none out of 10 for T. dimidiata (0%). Overall, the infestation index was 3.1% of households containing at least one triatomine; the crowding index was a mean of two Triatoma spp./household; and the colonization index was 0.38 for households based on presence of nymphs. Geographical distribution of triatomines in Oaxaca at the municipality level and endophilic behavior is also reported. Precise identification, endophilic habits, and infection rates of these triatomines are paramount for vector control programs of the Ministry of Health of Oaxaca and beyond.

Abstract Background Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, causes sudden death and chronic heart disease with no currently approved treatment. Objective To report epidemiologic and select cardiac characteristics associated with T. cruzi infection in dogs presenting to a teaching hospital in Texas. Animals Three hundred seventy‐five client‐owned dogs. Methods A retrospective search of medical records identified dogs tested for T. cruzi antibodies or with histologic T. cruzi parasites. Data retrieved included signalment, location of residence, reported reason for testing, cardiac troponin I (cTnI) concentration, and ECG abnormalities. Results Trypanosoma cruzi‐infected dogs (N = 63, 16.8%) were significantly younger than negative dogs (N = 312) (mean, 5.9 ± 3.8 versus 7.4 ± 4.0 years; P = .007) with no difference by sex or breed. Ninety‐one breeds were tested; the highest percent infected were non‐sporting (10/35; 29%) and toy breed (10/42; 24%) groups. The odds of infection were 13 times greater among dogs with an infected housemate or littermate (95% confidence interval [CI], 3.94‐50.45; P < .001). Infected dogs were more likely to have ventricular arrhythmias (odds ratio [OR], 2.19; 95% CI, 1.15‐4.33, P = .02), combinations of ECG abnormalities (OR, 2.91; 95% CI, 1.37‐5.99; P = .004), and cTnI >0.129 ng/mL (ADVIA; OR, 10.71; 95% CI, 1.60‐212.21; P = .035). Conclusions and Clinical Importance Dogs infected with T. cruzi were identified in Texas in many breed groups including breeds affected by well‐described heart diseases that mimic Chagas disease suggesting a need for increased awareness, including knowledge of when to consider testing.

Abstract Background Despite their importance as vectors of zoonotic parasites that can impact human and animal health, Culicoides species distribution across different habitat types is largely unknown. Here we document the community composition of Culicoides found in an urban environment including developed and natural sites in east central Texas, a region of high vector diversity due to subtropical climates, and report their infection status with haemoparasites. Results A total of 251 individual Culicoides were collected from May to June 2016 representing ten Culicoides species, dominated by C. neopulicaris followed by C. crepuscularis. We deposited 63 sequences to GenBank among which 25 were the first deposition representative for six Culicoides species: C. arboricola (n = 1); C. nanus (n = 4); C. debilipalpis (n = 2); C. haematopotus (n = 14); C. edeni (n = 3); and C. hinmani (n = 1). We also record for the first time the presence of C. edeni in Texas, a species previously known to occur in the Bahamas, Florida and South Carolina. The urban environments with natural area (sites 2 and 4) had higher species richness than sites more densely populated or in a parking lot (sites 1 and 3) although a rarefaction analysis suggested at least two of these sites were not sampled sufficiently to characterize species richness. We detected a single C. crepuscularis positive for Onchocercidae gen. sp. DNA and another individual of the same species positive for Haemoproteus sacharovi DNA, yielding a 2.08% prevalence (n = 251) for both parasites in this species. Conclusions We extend the knowledge of the Culicoides spp. community in an urban environment of Texas, USA, and contribute to novel sequence data for these species. Additionally, the presence of parasite DNA (Onchocercidae gen. sp. and H. sacharovi) from C. crepuscularis suggests the potential for this species to be a vector of these parasites.

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.

Tick-host bloodmeal associations are important factors when characterizing risks of associated pathogen transmission and applying appropriate management strategies. Despite their biological importance, comparatively little is known about soft tick (Argasidae) host associations in the United States compared to hard ticks (Ixodidae). In this study, we evaluated a PCR and direct Sanger sequencing method for identifying the bloodmeal hosts of soft ticks. We collected 381 cave-associated Ornithodoros turicata near San Antonio, Texas, USA, and also utilized eight colony-reared specimens fed artificially on known host blood sources over 1.5 years ago. We correctly identified the vertebrate host bloodmeals of two colony-reared ticks (chicken and pig) up to 1,105 days post-feeding, and identified bloodmeal hosts from 19 out of 168 field-collected soft ticks, including raccoon (78.9%), black vulture (10.5%), Texas black rattlesnake (5.3%), and human (5.3%). Our results confirm the retention of vertebrate blood DNA in soft ticks and advance the knowledge of argasid host associations in cave-dwelling O. turicata.

The ecology and host feeding patterns of many soft ticks (Ixodida: Argasidae) remain poorly understood. To address soft tick–host feeding associations, we fed Ornithodoros turicata Dugès on multiple host species and evaluated quantitative PCR (qPCR) and stable isotope analyses to identify the vertebrate species used for the bloodmeal. The results showed that a qPCR with host-specific probes for the cytochrome b gene successfully identified bloodmeals from chicken (Gallus gallus L.), goat (Capra aegagrus hircus L), and swine (Sus scrofa domesticus) beyond 330 days post-feeding and through multiple molting. Also, qPCR-based bloodmeal analyses could detect multiple host species within individual ticks that fed upon more than one species. The stable isotope bloodmeal analyses were based on variation in the natural abundance of carbon (13C/12C) and nitrogen (15N/14N) isotopes in ticks fed on different hosts. When compared to reference isotope signatures, this method discerned unique δ13C and δ15N signatures in the ticks fed on each host taxa yet could not discern multiple host species from O. turicata that fed on more than one host species. Given the significance of soft tick-borne zoonoses and animal diseases, elucidating host feeding patterns from field-collected ticks using these methods may provide insight for an ecological basis to disease management.

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.

In 2017, we surveyed forests in the upper Tennessee Valley, Tennessee, USA. We found Ixodes scapularis ticks established in 23 of 26 counties, 4 of which had Borrelia burgdorferi sensu stricto–infected ticks. Public health officials should be vigilant for increasing Lyme disease incidence in this region.

Northern pygmy mice from 2 localities in East Central Texas, USA, had proliferative epidermal lesions on the tail and feet. Electron microscopy of lesion tissue revealed poxvirus. Phylogenetic analyses indicated the virus differed 35% from its closest relatives, the Chordopoxvirinae. Future research is needed to determine whether this virus could affect human health.

Understanding the ecological and epidemiological roles of pets in the transmission of SARS-CoV-2 is critical for animal and human health, identifying household reservoirs, and predicting the potential enzootic maintenance of the virus. We conducted a longitudinal household transmission study of 76 dogs and cats living with at least one SARS-CoV-2-infected human in Texas and found that 17 pets from 25.6% of 39 households met the national case definition for SARS-CoV-2 infections in animals. This includes three out of seventeen (17.6%) cats and one out of fifty-nine (1.7%) dogs that were positive by RT-PCR and sequencing, with the virus successfully isolated from the respiratory swabs of one cat and one dog. Whole-genome sequences of SARS-CoV-2 obtained from all four PCR-positive animals were unique variants grouping with genomes circulating among people with COVID-19 in Texas. Re-sampling showed persistence of viral RNA for at least 25 d-post initial test. Additionally, seven out of sixteen (43.8%) cats and seven out of fifty-nine (11.9%) dogs harbored SARS-CoV-2 neutralizing antibodies upon initial sampling, with relatively stable or increasing titers over the 2–3 months of follow-up and no evidence of seroreversion. The majority (82.4%) of infected pets were asymptomatic. ‘Reverse zoonotic’ transmission of SARS-CoV-2 from infected people to animals may occur more frequently than recognized.

Great-tailed grackles (Quiscalus mexicanus) have dramatically expanded into North America over the past century. However, little is known about the blood that parasites they support. Here, for the first time, we document an assemblage of trypanosome, haemosporida, and filarial nematodes co-circulating in invasive great-tailed grackles. Between February and July, 2015, 61 individuals were captured in an urban environment of College Station, Texas. Field microscopy and molecular diagnostics indicate that 52% (24/46) were visually infected with filarioid nematodes, 24% (11/46) with avian trypanosomes, and 73% (n = 44/60) with haemosporida parasites, such as Haemoproteus (Parahaemoproteus) and Plasmodium cathemerium. Overall, 87% of great-tailed grackles were infected with blood parasites. Although 50% of individuals hosted parasites from multiple phylum, no patterns of parasite assembly were observed. Results indicate that great-tailed grackles can support a relatively high level of blood parasitism. However, the consequences for avian health remain to be determined.

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.

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.

Bird-facilitated introduction of ticks and associated pathogens is postulated to promote invasion of tick-borne zoonotic diseases into urban areas. Results of a longitudinal study conducted in suburban Chicago, Illinois, USA, during 2005–2010 show that 1.6% of 6,180 wild birds captured in mist nets harbored ticks. Tick species in order of abundance were Haemaphysalis leporispalustris, Ixodes dentatus, and I. scapularis, but 2 neotropical tick species of the genus Amblyomma were sampled during the spring migration. I. scapularis ticks were absent at the beginning of the study but constituted the majority of ticks by study end and were found predominantly on birds captured in areas designated as urban green spaces. Of 120 ticks, 5 were infected with Borrelia burgdorferi, spanning 3 ribotypes, but none were infected with Anaplasma phagocytophilum. Results allow inferences about propagule pressure for introduction of tick-borne diseases and emphasize the large sample sizes required to estimate this pressure.

Abstract Parasites and pathogens are increasingly recognized as significant drivers of ecological and evolutionary change in natural ecosystems. Concurrently, transmission of infectious agents among human, livestock, and wildlife populations represents a growing threat to veterinary and human health. In light of these trends and the scarcity of long‐term time series data on infection rates among vectors and reservoirs, the National Ecological Observatory Network (NEON) will collect measurements and samples of a suite of tick‐, mosquito‐, and rodent‐borne parasites through a continental‐scale surveillance program. Here, we describe the sampling designs for these efforts, highlighting sampling priorities, field and analytical methods, and the data as well as archived samples to be made available to the research community. Insights generated by this sampling will advance current understanding of and ability to predict changes in infection and disease dynamics in novel, interdisciplinary, and collaborative ways.