PATHOLOGY AND DISCRETE TYPING UNIT ASSOCIATIONS OF TRYPANOSOMA CRUZI INFECTION IN COYOTES (CANIS LATRANS) AND RACCOONS (PROCYON LOTOR) OF TEXAS, USA.

Trypanosoma cruzi is a vector-borne, protozoal parasite of mammals. Infected humans, dogs (Canis lupus familiaris), and nonhuman primates may remain asymptomatic or may develop Chagas disease, most commonly characterized by lymphoplasmacytic myocarditis with myocardial degeneration and fibrosis, ultimately resulting in heart failure. Although wildlife species have important roles as sylvatic reservoirs, investigations into the pathology of T. cruzi in wildlife are limited to a few studies documenting histologic lesions in opossums (Didelphis spp.) and raccoons (Procyon lotor). Pathology in coyotes (Canis latrans) has not, to our knowledge, been described, despite their recognition as a reservoir and close genetic relationship to domestic dogs. Our objectives were to perform a detailed, comparative cardiac pathology study of sympatric, naturally infected coyotes and raccoons, to characterize the overall T. cruzi infection prevalence in the heart and blood of each species via PCR, and to identify infecting discrete typing units (DTUs). We sampled hunter-harvested coyotes (n=120) and raccoons (n=24) in a 28 county region of central and south Texas, US. Raccoons were significantly more likely to have positive PCR results (P<0.001) with a prevalence of 62% (15/24), comprising DTU TcIV exclusively, with mild to no evidence of cardiac pathology. In contrast, coyotes had a lower infection prevalence (8%, 10/120), comprising DTU TcI exclusively, with lymphoplasmacytic myocarditis observed in four of the six PCR-positive animals. Many raccoons had PCR-positive blood and heart tissue simultaneously, supporting previous reports that raccoons maintain parasitemia into chronic stages of infection; in contrast, none of the PCR-positive coyotes were positive in both heart and blood. Our findings demonstrate marked differences in T. cruzi infection dynamics between coyotes and raccoons, with important implications for reservoir potential and their role in transmission cycles.

Seroprevalence for the tick-borne relapsing fever spirochete Borrelia turicatae among small and medium sized mammals of Texas.

BACKGROUND: In low elevation arid regions throughout the southern United States, Borrelia turicatae is the principal agent of tick-borne relapsing fever. However, endemic foci and the vertebrate hosts involved in the ecology of B. turicatae remain undefined. Experimental infection studies suggest that small and medium sized mammals likely maintain B. turicatae in nature, while the tick vector is a long-lived reservoir. METHODOLOGY/PRINCIPAL FINDINGS: Serum samples from wild caught rodents, raccoons, and wild and domestic canids from 23 counties in Texas were screened for prior exposure to B. turicatae. Serological assays were performed using B. turicatae protein lysates and recombinant Borrelia immunogenic protein A (rBipA), a diagnostic protein that is unique to RF spirochetes and may be a species-specific antigen. CONCLUSIONS/SIGNIFICANCE: Serological responses to B. turicatae were detected from 24 coyotes, one gray fox, two raccoons, and one rodent from six counties in Texas. These studies indicate that wild canids and raccoons were exposed to B. turicatae and are likely involved in the pathogen's ecology. Additionally, more work should focus on evaluating rodent exposure to B. turicatae and the role of these small mammals in the pathogen's maintenance in nature.

Prevalence and Seroprevalence of Trypanosoma cruzi Infection in a Military Population in Texas.

Recent biosurveillance findings at Joint Base San Antonio (JBSA), a large military installation located in south-central Texas, indicate the potential for vector-borne human Chagas disease. A cross-sectional study was conducted to determine the prevalence and seroprevalence of Trypanosoma cruzi infection in highest risk subpopulations on the installation, including students and instructors who work and sleep in triatomine-endemic field settings. Real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and indirect immunofluorescent antibody assay were performed on enrolled subjects (N = 1,033), none of whom tested positive for T. cruzi or anti-T. cruzi antibodies. Current countermeasures used during field training on JBSA appear to be sufficient for preventing autochthonous human Chagas disease.

A case of Chagas cardiomyopathy following infection in south central Texas.

Between 5 and 8 million people globally are infected with Trypanosoma cruzi, the causative parasitic agent of Chagas disease. The vast majority of incident infections originate in pockets of Latin America where domestic vector-borne transmission cycles are more common. Since 1955, when the first locally-acquired case was reported, fewer than 30 autochthonous cases have been documented in the United States. We describe the case of an 18-year-old US Air Force trainee, a native Texan with no travel history beyond the continental United States, who screened positive for T cruzi infection on blood donation and was subsequently found to have chronic Chagasic cardiomyopathy. This is the first documented case of Chagas disease in a US military trainee and one of the first known autochthonous cases of Chagasic cardiomyopathy in a Texas resident. Diagnostic, therapeutic, and military implications are discussed.

Diagnosis and Management of Borrelia turicatae Infection in Febrile Soldier, Texas, USA.

In August 2015, a soldier returned from field exercises in Texas, USA, with nonspecific febrile illness. Culture and sequencing of spirochetes from peripheral blood diagnosed Borrelia turicatae infection. The patient recovered after receiving doxycycline. No illness occurred in asymptomatic soldiers potentially exposed to the vector tick and prophylactically given treatment.

Trypanosoma cruzi Infection Prevalence and Bloodmeal Analysis in Triatomine Vectors of Chagas Disease From Rural Peridomestic Locations in Texas, 2013-2014.

Protozoan pathogen Trypanosoma cruzi (Chagas, 1909) is the etiologic agent of Chagas disease, which affects millions of people in Latin America. Recently, the disease has been gaining attention in Texas and the southern United States. Transmission cycle of the parasite involves alternating infection between insect vectors and vertebrate hosts (including humans, wildlife, and domestic animals). To evaluate vector T. cruzi parasite burden and feeding patterns, we tested triatomine vectors from 23 central, southern, and northeastern counties of Texas. Out of the 68 submitted specimens, the majority were genetically identified as Triatoma gerstaeckeri (Stal, 1859), with a few samples of Triatoma sanguisuga (LeConte, 1855), Triatoma lecticularia (Stal, 1859), Triatoma rubida (Uhler, 1894), and Triatoma protracta woodi (Usinger, 1939). We found almost two-thirds of the submitted insects were polymerase chain reaction-positive for T. cruzi Bloodmeal sources were determined for most of the insects, and 16 different species of mammals were identified as hosts. The most prevalent type of bloodmeal was human, with over half of these insects found to be positive for T. cruzi High infection rate of the triatomine vectors combined with high incidence of feeding on humans highlight the importance of Chagas disease surveillance in Texas. With our previous findings of autochthonous transmission of Chagas disease, urgent measures are needed to increase public awareness, vector control in and around homes, and Chagas screening of residents who present with a history of a triatomine exposure.

Assessment of the Geographic Distribution of Ornithodoros turicata (Argasidae): Climate Variation and Host Diversity.

BACKGROUND: Ornithodoros turicata is a veterinary and medically important argasid tick that is recognized as a vector of the relapsing fever spirochete Borrelia turicatae and African swine fever virus. Historic collections of O. turicata have been recorded from Latin America to the southern United States. However, the geographic distribution of this vector is poorly understood in relation to environmental variables, their hosts, and consequently the pathogens they transmit. METHODOLOGY: Localities of O. turicata were generated by performing literature searches, evaluating records from the United States National Tick Collection and the Symbiota Collections of Arthropods Network, and by conducting field studies. Maximum entropy species distribution modeling (Maxent) was used to predict the current distribution of O. turicata. Vertebrate host diversity and GIS analyses of their distributions were used to ascertain the area of shared occupancy of both the hosts and vector. CONCLUSIONS AND SIGNIFICANCE: Our results predicted previously unrecognized regions of the United States with habitat that may maintain O. turicata and could guide future surveillance efforts for a tick capable of transmitting high-consequence pathogens to human and animal populations.

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.

The Biology of the Triatomine Bugs Native to South Central Texas and Assessment of the Risk They Pose for Autochthonous Chagas Disease Exposure.

Triatomine bugs are a group of hematophagous arthropods that can serve as biological vectors for Trypanosoma cruzi , the etiological agent of American trypanosomiasis (Chagas disease). Because of differences in the biology and feeding habits among triatomine species, some are more likely than others to be involved in zoonotic and/or human-to-human transmission cycles of T. cruzi . In an attempt to assess the risk for Chagas disease exposure in south-central Texas, human habitations across Texas Health Service Region 8 (HSR 8) and surrounding counties were surveyed for triatomines to characterize the geographic distribution, species-specific biology, and T. cruzi -infection prevalence better. Between May 2010 and August 2013, a total of 545 triatomines representing all 5 known indigenous species (Triatoma gerstaeckeri, Triatoma indictiva, Triatoma lecticularia, Triatoma sanguisuga, and Triatoma protracta woodi) were collected from 59 sites across the region. Triatoma gerstaeckeri was the species most commonly found in domestic and peridomestic ecotopes across Texas HSR 8, representing over 80% of the triatomines collected. Adult T. gerstaeckeri exhibited a seasonal dispersal pattern that began in late April, peaked in mid-May, and then continued into August. On homes with available crevices in the exterior walls, adult T. gerstaeckeri were observed emerging from or entering these protective microhabitats, suggesting possible opportunistic colonization of some exterior walls compartments. Laboratory testing of triatomine hindgut contents for T. cruzi by PCR demonstrated the adult T. gerstaeckeri-infection prevalence across Texas HSR 8 to be 64%. Monitoring peridomestic adult T. gerstaeckeri over the seasonal dispersal peak demonstrated statistically significant increases in both their T. cruzi -infection prevalence (P < 0.01) and tendency to invade human dwellings (P < 0.01) in the later aspect of the emergence peak. In addition to the adult insects, variably sized and staged nymphs were recovered from the inside of 6 separate homes across Texas HSR 8. The results of this study show that T. gerstaeckeri is a widespread and common triatomine species across Texas HSR 8 and documented it to have some notable synanthropic tendencies. The high prevalence of T. cruzi infection in native triatomines, and the high frequency with which T. gerstaeckeri is recovered from human habitations, suggests that there is a risk for human exposure to T. cruzi in Texas HSR 8. Because of this, Chagas disease should be considered on the list of differential diagnoses for cases of cardiac arrhythmia, dilated cardiomyopathy, or heart failure in south-central Texas.

Ball python nidovirus: a candidate etiologic agent for severe respiratory disease in Python regius.

UNLABELLED: A severe, sometimes fatal respiratory disease has been observed in captive ball pythons (Python regius) since the late 1990s. In order to better understand this disease and its etiology, we collected case and control samples and performed pathological and diagnostic analyses. Electron micrographs revealed filamentous virus-like particles in lung epithelial cells of sick animals. Diagnostic testing for known pathogens did not identify an etiologic agent, so unbiased metagenomic sequencing was performed. Abundant nidovirus-like sequences were identified in cases and were used to assemble the genome of a previously unknown virus in the order Nidovirales. The nidoviruses, which were not previously known to infect nonavian reptiles, are a diverse order that includes important human and veterinary pathogens. The presence of the viral RNA was confirmed in all diseased animals (n = 8) but was not detected in healthy pythons or other snakes (n = 57). Viral RNA levels were generally highest in the lung and other respiratory tract tissues. The 33.5-kb viral genome is the largest RNA genome yet described and shares canonical characteristics with other nidovirus genomes, although several features distinguish this from related viruses. This virus, which we named ball python nidovirus (BPNV), will likely establish a new genus in Torovirinae subfamily. The identification of a novel nidovirus in reptiles contributes to our understanding of the biology and evolution of related viruses, and its association with lung disease in pythons is a promising step toward elucidating an etiology for this long-standing veterinary disease. IMPORTANCE: Ball pythons are popular pets because of their diverse coloration, generally nonaggressive behavior, and relatively small size. Since the 1990s, veterinarians have been aware of an infectious respiratory disease of unknown cause in ball pythons that can be fatal. We used unbiased shotgun sequencing to discover a novel virus in the order Nidovirales that was present in cases but not controls. While nidoviruses are known to infect a variety of animals, this is the first report of a nidovirus recovered from any reptile. This report will enable diagnostics that will assist in determining the role of this virus in the causation of disease, which would allow control of the disease in zoos and private collections. Given its evolutionary divergence from known nidoviruses and its unique host, the study of reptile nidoviruses may further our understanding of related diseases and the viruses that cause them in humans and other animals.

Immunohistochemical detection of a unique protein within cells of snakes having inclusion body disease, a world-wide disease seen in members of the families Boidae and Pythonidae.

Inclusion body disease (IBD) is a worldwide disease in captive boa constrictors (boa constrictor) and occasionally in other snakes of the families Boidae and Pythonidae. The exact causative agent(s) and pathogenesis are not yet fully understood. Currently, diagnosis of IBD is based on the light microscopic identification of eosinophilic intracytoplasmic inclusion bodies in hematoxylin and eosin stained tissues or blood smears. An antigenically unique 68 KDa protein was identified within the IBD inclusion bodies, called IBD protein. A validated immuno-based ante-mortem diagnostic test is needed for screening snakes that are at risk of having IBD. In this study, despite difficulties in solubilizing semi-purified inclusion bodies, utilizing hybridoma technology a mouse anti-IBD protein monoclonal antibody (MAB) was produced. The antigenic specificity of the antibody was confirmed and validated by western blots, enzyme-linked immunosorbent assay, immuno-transmission electron microscopy, and immunohistochemical staining. Paraffin embedded tissues of IBD positive and negative boa constrictors (n=94) collected from 1990 to 2011 were tested with immunohistochemical staining. In boa constrictors, the anti-IBDP MAB had a sensitivity of 83% and specificity of 100% in detecting IBD. The antibody also cross-reacted with IBD inclusion bodies in carpet pythons (Morelia spilota) and a ball python (python regius). This validated antibody can serve as a tool for the development of ante-mortem immunodiagnostic tests for IBD.

Snakebite injuries treated in United States emergency departments, 2001-2004.

OBJECTIVE: Venomous and nonvenomous snakes are found throughout most of the United States. While the literature on treatment is robust, there is not a current national epidemiologic profile of snakebite injuries in the United States. National estimates of such injuries treated in emergency departments (EDs) are presented along with characteristics of the affected population. METHODS: Data on snakebite injuries were abstracted from the National Electronic Injury Surveillance System-All Injury Program (2001-04). Variables included age, gender, body part affected, cause, disposition, and treatment date. When available, location, intentionality of the interaction, and snake species were coded based on narrative comments. Estimates were weighted and analyzed with SPSS Complex Samples. RESULTS: An estimated 9873 snakebites were treated in US EDs each year between 2001 and 2004. Males were more frequently seen in the ED for snakebites than were females (males: 72.0% [95% confidence interval (CI), 68.0-75.7]; females: 28.0% [95% CI, 24.3-32.0]). Approximately 32% of patients were known to be bitten by venomous species. Overall, more than one quarter of patients were hospitalized (27.9% [95% CI, 15.9-44.2]), although 58.9% of patients with known venomous bites were hospitalized (95% CI, 41.5-74.3). CONCLUSIONS: While they are rare events, snakebites cause nearly 10,000 visits to EDs for treatment every year. Epidemiologic data regarding snakebites provide practicing physicians with an understanding of the population affected and can help guide public health practitioners in their prevention efforts.

Venomous adversaries: a reference to snake identification, field safety, and bite-victim first aid for disaster-response personnel deploying into the hurricane-prone regions of North America.

Each hurricane season, emergency-preparedness deployment teams including but not limited to the Office of Force Readiness and Deployment of the US Public Health Service, Federal Emergency Management Agency, Deployment Medical Assistance Teams, Veterinary Medical Assistance Teams, and the US Army and Air Force National Guard are at risk for deploying into hurricane-stricken areas that harbor indigenous hazards, including those posed by venomous snakes. North America is home to 2 distinct families of venomous snakes: 1) Viperidae, which includes the rattlesnakes, copperheads, and cottonmouths; and 2) Elapidae, in which the only native species are the coral snakes. Although some of these snakes are easily identified, some are not, and many rank among the most feared and misunderstood animals. This article specifically addresses all the native species of venomous snakes that inhabit the hurricane-prone regions of North America and is intended to serve as a reference to snake identification, basic field safety procedures, and the currently recommended first-aid measures for snakebite casualties.

Wide-band spectral tuning of heat receptors in the pit organ of the copperhead snake (Crotalinae).

Receptors located in the facial pit organ of certain species of snake signal the presence of prey. Infrared radiation is an effective stimulus suggesting that these receptors may be low-threshold temperature receptors. We recorded from the nerve innervating the pit organ of snakes belonging to the family of Crotalinae while stimulating the receptive area with well-defined optical stimuli. The objective was to determine the sensitivity of these receptors to a wide range (0.400-10.6 micro m) of optical stimuli to determine if a temperature-sensitive or photosensitive protein initiated signal transduction. We found that receptors in the pit organ exhibited a unique broad response to a wide range of electromagnetic radiation ranging from the near UV to the infrared. The spectral tuning of these receptors parallels closely the absorption spectra of water and oxyhemoglobin, the predominant chromophore in tissue. Our results support the hypothesis that these are receptors activated by minute temperature changes induced by direct absorption of optical radiation in the thin pit organ membrane.