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Objective: To describe associations between cardiac abnormalities and Trypanosoma cruzi serostatus by use of a simplified diagnostic evaluation in dogs at risk for T cruzi infection., Methods: A prospective, cross-sectional study was performed using a simplified diagnostic evaluation including high-sensitivity cardiac troponin I, 30-second ECG, and echocardiogram with 7 variables in 46 client-owned dogs from high-risk environments. Dogs were categorized as serologically positive (SP), negative (SN), or discordant (SD) by use of 2 antibody tests. Functional evaluation of cardiac health scores and blood PCR were obtained., Results: Dogs were SP (n = 19), SN (17), and SD (10), with 9 PCR positive (7 SP, 1 SN, 1 SD). Troponin was above reference range in 6 of 46 (4 SP, 1 SN, 1 SD), and functional evaluation of cardiac health scores were 0 in all dogs. Conduction system abnormalities (prolonged interval durations, second-degree atrioventricular block, splintered QRS complex) and ventricular arrhythmias were documented in 8 (7 SP, 0 SN, 1 SD). Twenty-six (12 SP, 8 SN, 6 SD) had echocardiographic abnormalities, most often myxomatous mitral valve disease (MMVD) and left ventricular enlargement. Seropositive dogs were significantly older and had a higher likelihood of MMVD. Conduction system abnormalities were associated with positive serostatus., Conclusions: Echocardiographic abnormalities were complicated by MMVD and did not distinguish between serostatus. An ECG with assessment and detailed measurement of complexes and cardiac troponin I are simple tests to perform with abnormalities detected in seroreactive dogs., Clinical Relevance: Electrocardiographic abnormalities in high-risk or seroreactive dogs should prompt further evaluation and monitoring of T cruzi infection.

The Mexican free-tailed bat (Tadarida brasiliensis) is one of the most abundant mammals in North America. Mexican free-tailed bats have a wide geographic range stretching from northern South America to the western United States. Bats are theorized to be the original hosts for Trypanosoma cruzi -the causative agent of Chagas disease- and can serve as a source of infection to triatomine insect vectors that feed upon them. Chagas disease is a neglected tropical disease across the Americas where triatomines are present, including the southern United States, where Texas reports this highest number of locally-acquired human cases. To learn more about the role of bats in the ecology of Chagas disease in Texas, we surveyed a colony of Mexican free-tailed bats from Brazos County, Texas, for T. cruzi using carcasses salvaged after an extreme weather event. A total of 283 Mexican free-tailed bats collected in February 2021 were dissected and DNA from the hearts and kidneys was used for T. cruzi detection via qPCR. None of the bat hearts or kidneys tested positive for T. cruzi; this sample size affords 95% confidence that the true prevalence of T. cruzi in this population does not exceed 1%. Future sampling of multiple bat species as well as migrant and resident colonies of Mexican free-tailed bats across different times of the year over a broader geographic range would be useful in learning more about the role of bats in the ecology of Chagas disease in Texas., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests. Ilana Mosley reports financial support was provided by National Institutes of Health. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Triatomine insects are vectors of the protozoan parasite Trypanosoma cruzi- the causative agent of Chagas disease. Chagas disease is endemic to Latin America and the southern United States and can cause severe cardiac damage in infected mammals, ranging from chronic disease to sudden death. Identifying interactions among triatomines, T. cruzi discrete typing units (DTUs), and blood feeding hosts is necessary to understand parasite transmission dynamics and effectively protect animal and human health. Through manual insect trapping efforts, kennel staff collections, and with the help of a trained scent detection dog, we collected triatomines from 10 multi-dog kennels across central and south Texas over a one-year period (2018-2019) and tested a subset to determine their T. cruzi infection status and identify the primary bloodmeal hosts. We collected 550 triatomines, including Triatoma gerstaeckeri (n = 515), Triatoma lecticularia (n = 15), Triatoma sanguisuga (n = 6), and Triatoma indictiva (n = 2), with an additional 10 nymphs and 2 adults unable to be identified to species. The trained dog collected 42 triatomines, including nymphs, from areas not previously considered vector habitat by the kennel owners. Using qPCR, we found a T. cruzi infection prevalence of 47 % (74/157), with T. lecticularia individuals more likely to be infected with T. cruzi than other species. Infected insects harbored two T. cruzi discrete typing units: TcI (64 %), TcIV (23 %), and mixed TcI/TcIV infections (13 %). Bloodmeal host identification was successful in 50/149 triatomines, revealing the majority (74 %) fed on a dog (Canis lupus), with other host species including humans (Homo sapiens), raccoons (Procyon lotor), chickens (Gallus gallus), wild pig (Sus scrofa), black vulture (Coragyps atratus), cat (Felis catus), and curve-billed thrasher (Toxostoma curviostre). Given the frequency of interactions between dogs and infected triatomines in these kennel environments, dogs may be an apt target for future vector control and T. cruzi intervention efforts., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Trypanosoma cruzi , the causative agent of Chagas disease, is a zoonotic, vector-borne, protozoan hemoflagellate with a wide host range. An 11-yr-old, captive-bred male De Brazza's monkey ( Cercopithecus neglecus ) presented with weight loss despite normal appetite. Examination revealed hypoglycemia, nonregenerative anemia, and many trypanosomes on a blood smear. A whole blood sample was PCR-positive for T. cruzi discrete typing unit TcIV and the monkey seroconverted using two different methods. The monkey was treated with the standard human dose of benznidazole twice daily for 60 d; however, blood obtained over the next 1.5 yr posttreatment remained PCR-positive for T. cruzi . A second course of benznidazole at a higher dose but lower frequency for 26 wk was required for the monkey to convert to sustained PCR-negative status. The monkey recovered with no apparent lasting effects.

Trypanosoma cruzi naturally infects a broad range of mammalian species and frequently results in the pathology that has been most extensively characterized in human Chagas disease. Currently employed treatment regimens fail to achieve parasitological cure of T. cruzi infection in the majority of cases. In this study, we have extended our previous investigations of more effective, higher dose, intermittent administration protocols using the FDA-approved drug benznidazole (BNZ), in experimentally infected mice and in naturally infected dogs and nonhuman primates (NHP). Collectively, these studies demonstrate that twice-weekly administration of BNZ for more than 4 months at doses that are ~2.5-fold that of previously used daily dosing protocols, provided the best chance to obtain parasitological cure. Dosing less frequently or for shorter time periods was less dependable in all species. Prior treatment using an ineffective dosing regimen in NHPs did not prevent the attainment of parasitological cure with an intensified BNZ dosing protocol. Furthermore, parasites isolated after a failed BNZ treatment showed nearly identical susceptibility to BNZ as those obtained prior to treatment, confirming the low risk of induction of drug resistance with BNZ and the ability to adjust the treatment protocol when an initial regimen fails. These results provide guidance for the use of BNZ as an effective treatment for T. cruzi infection and encourage its wider use, minimally in high value dogs and at-risk NHP, but also potentially in humans, until better options are available., Competing Interests: The authors declare no conflict of interest.

American trypanosomiasis is caused by the zoonotic protozoa Trypanosoma cruzi and primarily results in heart disease. Organisms also infect the central nervous system (CNS). The Texas A&M University veterinary teaching hospital archive was searched for dogs with CNS disease with intralesional protozoal amastigotes. This study summarizes 4 cases of dogs with disseminated trypanosomiasis and CNS involvement confirmed by quantitative polymerase chain reaction (qPCR) with T. cruzi primers. Clinical signs included lethargy, respiratory distress, tetraparesis, and seizures. Central nervous system lesions included meningeal congestion (1/4), necrosis with hemorrhage in the spinal cord gray and white matter (2/4), and histiocytic meningoencephalitis (4/4), and meningomyelitis (2/4) with intralesional and intracellular protozoal. Genotyping identified 1 case of T. cruzi discrete typing unit (DTU) TcI and 2 cases as TcIV, both are common variants in the United States. Trypanosomiasis should be considered a differential diagnosis for dogs with CNS signs in T. cruzi -endemic areas.

Objective: To characterize clinical and epidemiologic features of SARS-CoV-2 in companion animals detected through both passive and active surveillance in the US., Animals: 204 companion animals (109 cats, 95 dogs) across 33 states with confirmed SARS-CoV-2 infections between March 2020 and December 2021., Procedures: Public health officials, animal health officials, and academic researchers investigating zoonotic SARS-CoV-2 transmission events reported clinical, laboratory, and epidemiologic information through a standardized One Health surveillance process developed by the CDC and partners., Results: Among dogs and cats identified through passive surveillance, 94% (n = 87) had reported exposure to a person with COVID-19 before infection. Clinical signs of illness were present in 74% of pets identified through passive surveillance and 27% of pets identified through active surveillance. Duration of illness in pets averaged 15 days in cats and 12 days in dogs. The average time between human and pet onset of illness was 10 days. Viral nucleic acid was first detected at 3 days after exposure in both cats and dogs. Antibodies were detected starting 5 days after exposure, and titers were highest at 9 days in cats and 14 days in dogs., Clinical Relevance: Results of the present study supported that cats and dogs primarily become infected with SARS-CoV-2 following exposure to a person with COVID-19, most often their owners. Case investigation and surveillance that include both people and animals are necessary to understand transmission dynamics and viral evolution of zoonotic diseases like SARS-CoV-2.

Trypanosoma cruzi naturally infects a wide variety of wild and domesticated mammals, in addition to humans. Depending on the infection dose and other factors, the acute infection can be life-threatening, and in all cases, the risk of chagasic heart disease is high in persistently infected hosts. Domestic, working, and semi-feral dogs in the Americas are at significant risk of T. cruzi infection and in certain settings in the southern United States, the risk of new infections can exceed 30% per year, even with the use of vector control protocols. In this study, we explored whether intermittent low-dose treatment with the trypanocidal compound benznidazole (BNZ) during the transmission season, could alter the number of new infections in dogs in an area of known, intense transmission pressure. Preliminary studies in mice suggested that twice-weekly administration of BNZ could prevent or truncate infections when parasites were delivered at the mid-point between BNZ doses. Pre-transmission season screening of 126 dogs identified 53 dogs (42.1%) as T. cruzi infection positive, based upon blood PCR and Luminex-based serology. Serial monitoring of the 67 uninfected dogs during the high transmission season (May to October) revealed 15 (22.4%) new infections, 6 in the untreated control group and 9 in the group receiving BNZ prophylaxis, indicating no impact of this prophylaxis regimen on the incidence of new infections. Although these studies suggest that rigorously timed and more potent dosing regimen may be needed to achieve an immediate benefit of prophylaxis, additional studies would be needed to determine if drug prophylaxis reduced disease severity despite this failure to prevent new infections., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Bustamante et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Flies and other arthropods mechanically transmit multiple pathogens and a recent experimental study demonstrated house flies, Musca domestica L. (Diptera: Muscidae), can mechanically transmit SARS-CoV-2. The purpose of this study was to explore the possibility of mechanical transmission of SARS-CoV-2 by domestic insects and their potential as a xenosurveillance tool for detection of the virus. Flies were trapped in homes where at least one confirmed human COVID-19 case(s) resided using sticky and liquid-baited fly traps placed inside and outside the home in the Texas counties of Brazos, Bell, and Montgomery, from June to September 2020. Flies from sticky traps were identified, pooled by taxa, homogenized, and tested for the presence of SARS-CoV-2 RNA using quantitative reverse transcription PCR (RT-qPCR). Liquid traps were drained, and the collected fluid similarly tested after RNA concentration. We processed the contents of 133 insect traps from 40 homes, which contained over 1,345 individual insects of 11 different Diptera families and Blattodea. These individuals were grouped into 243 pools, and all tested negative for SARS-CoV-2 RNA. Fourteen traps in seven homes were deployed on the day that cat or dog samples tested positive for SARS-CoV-2 RNA by nasal, oral, body, or rectal samples. This study presents evidence that biting and nonbiting flies and cockroaches (Blattodea) are not likely to contribute to mechanical transmission of SARS-CoV-2 or be useful in xenosurveillance for SARS-CoV-2., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America.)

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., (© 2022. The Author(s).)

As part of a longitudinal household transmission study of pets living with persons with COVID-19 in Texas, two pets were confirmed to be infected with the SARS-CoV-2 B.1.1.7 variant of concern (VOC). The pets were a dog and a cat from the same household, sampled two days after their owner tested positive for COVID-19. The oral, nasal and fur swabs for both pets tested positive for SARS-CoV-2 by qRT-PCR and consensus whole-genome sequences from the dog and cat were 100% identical and matched the B.1.1.7 VOC. Virus was isolated from the cat's nasal swab. One month after initial detection of infection, the pets were re-tested twice at which time only the fur swabs (both pets) and oral swab (dog only) remained positive, and neutralizing antibodies for SARS-CoV-2 were present in both animals. Sneezing by both pets was noted by the owner in the weeks between initial and follow-up testing. This study documents the first detection of B.1.1.7. in companion animals in the United States, and the first genome recovery and isolation of B.1.1.7 variant of concern globally in any animal., (© 2021 Wiley-VCH GmbH.)

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 (PRNT 90 ), 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.

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., Competing Interests: The authors have declared that no competing interests exist.

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.

Trypanosoma cruzi is a zoonotic protozoan parasite vectored by triatomine insects that are endemic to the Americas, including the southern United States. Surveillance of domestic dogs for T. cruzi exposure allows for the determination of geographic regions of transmission that are relevant for human and animal health. The U.S. Department of Homeland Security (DHS) working dogs provide critical security and detection services across the country, and many train or work in the southern United States, where they are at risk for T. cruzi exposure. We sampled blood from 1,610 working dogs (predominantly Belgian Malinois, German shepherds, and Labrador retrievers) from six task forces (including the Transportation Security Administration, Customs and Border Protection, Secret Service, and more) and two canine training centers across 41 states from 2015 to 2018. Canine sera that were reactive on at least two independent serological assays were considered positive for anti- T.-cruzi antibodies. In addition, up to three independent polymerase chain reaction (PCR) assays were used to detect and type T. cruzi DNA. Overall seroprevalence was 7.5%, and four dogs (0.25%, n = 1,610) had detectable parasite DNA in the blood, comprising parasite discrete taxonomic units (DTUs) TcIV and a coinfection of TcI/TcIV. Dogs that worked within versus outside of the geographic range of established triatomines showed comparable seroprevalence (7.3% and 9.2%, respectively; P = 0.61). Determining the prevalence of T. cruzi in these working dogs and looking at spatially associated risk factors have practical implications for disease risk management and could assist with improved control measures to protect both animal and human health.

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 < 0.0001), and positive and discordant dogs had a higher mean number of different types of ECG abnormalities than negative dogs (p < 0.001-0.014). The most common cardiac abnormalities included supraventricular and ventricular arrhythmias and atrioventricular block. Positive dogs had higher serum concentrations of cTnI than both negative dogs (p = 0.044) and discordant dogs (p = 0.06). Based on dog handler reports, nearly all (4/5; 80%) dogs with reported performance decline or fatigue were T. cruzi-infected dogs., Conclusions: Further understanding cardiac manifestations in dogs naturally infected with T. cruzi is critical for prognostication, establishing a baseline for drug and vaccine studies, and better understanding of zoonotic risk.

As tick-borne diseases continue to emerge across the United States, there is need for a better understanding of the tick and pathogen communities in the southern states and of habitat features that influence transmission risk. We surveyed questing and on-host ticks in pine-dominated forests with various fire management regimes in the Sam Houston National Forest, a popular recreation area near Houston, Texas. Four linear transects were established- two with a history of controlled burns, and two unburned. Systematic drag sampling yielded 112 ticks from two species, Ixodes scapularis (n=73) and Amblyomma americanum (n=39), with an additional 106 questing ticks collected opportunistically from drag cloth operators. There was a significant difference in systematically-collected questing tick density between unburned (15 and 18 ticks/1000 m 2 ) and burned (2 and 4 ticks/1000 m 2 ) transects. We captured 106 rodents and found 74 ticks on the rodents, predominantly Dermacentor variabilis. One unburned transect had significantly more ticks per mammal than any of the other three transects. DNA of Rickettsia species was detected in 146/292 on and off-host ticks, including the 'Rickettsial endosymbiont of I. scapularis' and Rickettsia amblyommatis, which are of uncertain pathogenicity to humans. Borrelia lonestari was detected in one A. americanum, while Borrelia burgdorferi sensu stricto, the agent of Lyme disease, was not detected in any tick samples. Neither Borrelia nor Rickettsia spp. were detected in any of the mammal ear biopsies (n=64) or blood samples (n=100) tested via PCR. This study documents a high prevalence in ticks of Rickettsia spp. thought to be endosymbionts, a low prevalence of relapsing fever group Borrelia in ticks, and a lack of detection of Lyme disease-group Borrelia in both ticks and mammals in an east Texas forested recreation area. Additionally, we observed low questing tick density in areas with a history of controlled burns. These results expand knowledge of tick-borne disease ecology in east Texas which can aid in directing future investigative, modeling, and management efforts., (Copyright © 2019 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Changing climatic conditions and the northward expansion of ticks and pathogens are of immense importance to human, animal, and environmental health assessment and risk management. From 2014 through 2015, a wildlife rehabilitation center in south-central Arizona, US noted soft ticks (Argasidae) infesting 23 birds, including Cooper's Hawks ( Accipiter cooperii ), Gray Hawks ( Buteo plagiatus ), Harris's Hawks ( Parabuteo unicinctus ), Red-tailed Hawks ( Buteo jamaicensis ), Great Horned Owls ( Bubo virginianus ), Common Ravens ( Corvus corax ), and a Greater Roadrunner ( Geococcyx californianus ), during the late summer seasonal rainy seasons. The parasites numbered in the hundreds on individual birds. Infested birds were moribund, obtunded, or paralyzed on presentation, with no prior histories of illness or evidence of trauma. Tick and avian blood samples were collected for vector-borne pathogen analysis focusing on the molecular detection of Rickettsia and Borrelia species. Ticks were identified as the neotropical species of soft tick, Argas ( Persicargas ) giganteus ; their occurrence in the southern US on raptors represented an expansion of host range. Pathogen testing of ticks showed that 41% of 54 ticks were infected with Rickettsia hoogstraalii and 23% of 26 ticks were infected with a relapsing fever Borrelia ; both agents are associated with uncertain health consequences. Among the blood samples, one was infected with the same Borrelia spp.; this Red-tailed Hawk also had Borrelia- positive ticks. With supportive therapy, the majority of birds, 74% (17/23), recovered and were released or permanent residents. The management of soft tick-infested birds and mitigation of future disease risk will require additional characterizations of these poorly studied soft ticks and their associated pathogens.

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., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Ixodes scapularis (Acari: Ixodidae) is the principal vector of Borrelia burgdorferi (the etiologic agent of Lyme disease) in the eastern and midwestern United States. Recent efforts have documented the first established population of I. scapularis in South Dakota, representing a western expansion of the known species distribution. Our goal was to describe the current distribution of I. scapularis in eastern South Dakota and to survey for the presence of B. burgdorferi in questing I. scapularis. We surveyed for the presence of adult and nymphal I. scapularis in seven counties within South Dakota, including 13 locales from 2016 to 2017. We then tested all I. scapularis, including those collected in 2015 from a previous study, for the presence of B. burgdorferi sensu stricto, via quantitative and conventional polymerase chain reaction. Here, we document the presence of I. scapularis in four new counties in South Dakota, and report the first instance of B. burgdorferi in a questing tick in South Dakota. Coupled with data from the Centers for Disease Control and Prevention and Companion Animal Parasite Council our data show that the risk of contracting Lyme disease in South Dakota is low, but existent and should be an important consideration with regard to public health, pets, and wildlife.

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.

In 2006, Nabity et al. reported on an atypical presentation of Trypanosoma cruzi (T. cruzi) infection in an 8-month old English Mastiff from central Texas. Clinical signs and laboratory findings included lymphadenopathy, weight loss, amastigotes in lymph node aspirates, and initial serological results suggestive of either T. cruzi or Leishmania infection. Given the poor prognosis, the dog was euthanized and subsequent testing and culture of parasites from a lymph node revealed T. cruzi infection. Because different parasite discrete typing units (DTUs) are potentially associated with different disease outcomes in a variety of mammalian hosts, an understanding of these relationships in naturally infected dogs may be useful for informing canine prognosis and may also have human health implications. Here, we compared strains using culture versus culture-independent methods. We subjected archived cultured parasites harvested from the lymph node in the infected Mastiff to two independent approaches for determining parasite DTU, including sequencing of the TcSC5D gene and use of DTU-specific qPCR probes to hybridize the nuclear spliced leader intergenic region (SL-IR). Both approaches revealed T. cruzi discrete typing unit TcIV. Testing of multiple other tissues directly without culturing, including heart/tongue, intestine, trachea/lymph nodes, and uterus/ovary, provided further evidence of disseminated TcIV infection in this dog. We report T. cruzi DTU TcIV as the cause of a severe disseminated infection in a dog from an area with triatomine vectors in central Texas, adding to the limited body of clinicopathologic data that links specific parasite strains to disease outcomes in dogs in the US. Future studies to type parasites from asymptomatic dogs and those with diverse disease manifestations will be useful in informing the degree to which parasite genetics is associated with disease presentation and severity. If applied to antemortem samples, diagnostic typing of parasites from infected dogs may assist in determining prognosis., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)

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.

Alzheimer's disease (AD) is characterized by neuronal death, neurofibrillary tangles, and senile plaques. Amyloid-beta (Aβ) is the major component of plaques and consists of two prominent isoforms, Aβ40 and Aβ42. As many risk factors for AD are vascular in origin and blood vessel defects in clearing Aβ from the brain are a potential key component of AD pathology, we have focused on the neuron-blood vessel interface, and in particular, the vascular basement membrane, which coats blood vessels and physically separates them from neurons. A prominent component of the vascular basement membrane is the extracellular matrix proteoglycan perlecan. Domain V (DV) is the C-terminal domain and is generated by perlecan proteolysis. DV interacts with the α2 integrin and Aβ is a ligand for both α2β1 and αvβ1. Due to the known interaction of DV with α2β1 and α2β1's requirement for Aβ deposition and neurotoxicity, we hypothesized that DV and/or its C-terminal domain, LG3, might alter neurotoxic signaling pathways by directly blocking or otherwise interfering with α2β1 binding by Aβ. Our study suggests that α2β1 mediates Aβ-induced activation of c-Jun and caspase-3, key components of the neurotoxic pathway, in primary cortical and hippocampal neurons. We further demonstrate that DV and/or LG3 may therapeutically modulate these α2β1 mediated neurotoxic effects suggesting that they or other α2β1 integrin modulators could represent a novel approach to treat AD. Finally, our results suggest different neurotoxicity susceptibilities between cortical and hippocampal neurons to Aβ40 and Aβ42 as further underscored by differing neuroprotective potencies of LG3 in each cell type.

Background: Marking wild mosquitoes is important for understanding their ecology, behaviours and role in disease transmission. Traditional insect marking techniques include using fluorescent dyes, protein labels, radioactive labels and tags, but such techniques have various limitations; notably low marker retention and inability to mark wild mosquitoes at source. Stable isotopes are gaining wide spread use for non-invasive marking of arthropods, permitting greater understanding of mosquito dispersal and responses to interventions. We describe here a simple technique for marking naturally-breeding malaria and dengue vectors using stable isotopes of nitrogen (15N) and carbon (13C), and describe potential field applications., Methods: We created man-made aquatic mosquito habitats and added either 15N-labelled potassium nitrate or 13C-labelled glucose, leaving non-adulterated habitats as controls. We then allowed wild mosquitoes to lay eggs in these habitats and monitored their development in situ. Pupae were collected promptly as they appeared and kept in netting cages. Emergent adults (in pools of ~4 mosquitoes/pool) and individually stored pupae were desiccated and analysed using Isotope Ratio Mass Spectrometry (IRMS)., Findings: Anopheles gambiae s.l and Aedes spp. from enriched 13C and enriched 15N larval habitats had significantly higher isotopic levels than controls (P = 0.005), and both isotopes produced sufficient distinction between marked and unmarked mosquitoes. Mean δ15N for enriched females and males were 275.6±65.1 and 248.0±54.6, while mean δ15N in controls were 2.1±0.1 and 3.9±1.7 respectively. Similarly, mean δ13C for enriched females and males were 36.08±5.28 and 38.5±6.86, compared to -4.3±0.2 and -7.9±3.6 in controls respectively. Mean δ15N and δ13C was significantly higher in any pool containing at least one enriched mosquito compared to pools with all unenriched mosquitoes, P<0.001. In all cases, there were variations in standardized isotopic ratios between mosquito species., Conclusion: Enrichment of semi-natural mosquito larval habitats with stable isotopes of nitrogen and carbon resulted in effective marking of Anopheles and Aedes mosquitoes colonizing these habitats. This approach can significantly enhance studies on mosquito eco-physiology, dispersal, pathogen transmission and responses to control measures.

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., (Copyright © 2015 Elsevier B.V. All rights reserved.)

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.

Migratory birds have the potential to transport exotic vectors and pathogens of human and animal health importance across vast distances. We systematically examined birds that recently migrated to the United States from the Neotropics for ticks. We screened both ticks and birds for tick-borne pathogens, including Rickettsia species and Borrelia burgdorferi. Over two spring seasons (2013 and 2014), 3.56% of birds (n = 3,844) representing 42.35% of the species examined (n = 85) were infested by ticks. Ground-foraging birds with reduced fuel stores were most commonly infested. Eight tick species were identified, including seven in the genus Amblyomma, of which only Amblyomma maculatum/Amblyomma triste is known to be established in the United States. Most ticks on birds (67%) were neotropical species with ranges in Central and South America. Additionally, a single Ixodes genus tick was detected. A total of 29% of the ticks (n = 137) and no avian blood samples (n = 100) were positive for infection with Rickettsia species, including Rickettsia parkeri, an emerging cause of spotted fever in humans in the southern United States, a species in the group of Rickettsia monacensis, and uncharacterized species and endosymbionts of unknown pathogenicity. No avian tick or blood samples tested positive for B. burgdorferi, the etiologic agent of Lyme disease. An extrapolation of our findings suggests that anywhere from 4 to 39 million exotic neotropical ticks are transported to the United States annually on migratory songbirds, with uncertain consequences for human and animal health if the current barriers to their establishment and spread are overcome., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Identification of the vertebrate hosts upon which hematophagous arthropods feed provides key information for understanding the ecology and transmission of vector-borne diseases. Bloodmeal analysis of ticks presents unique challenges relative to other vectors, given the long interval between bloodmeal acquisition and host-seeking, during which DNA degradation occurs. This study evaluates DNA-based and stable isotope-based bloodmeal analysis methodologies for the lone star tick, Amblyomma americanum (Linneaus, 1758), in an experimental study with chicken as the known host. We subjected ticks of different ages and environmental rearing conditions to three DNA-based approaches and a stable isotopic analysis, which relies on the natural variation of nitrogen ((15)N/(14)N) and carbon ((13)C/(12)C) isotopes. While all three DNA-based approaches were successful in identifying the bloodmeal host of the engorged nymphs, only the probe-based RT-PCR was able to detect host DNA in aged ticks, the success of which was low and inconsistent across age and rearing treatments. In contrast, the stable isotope analysis showed utility in determining the host across all ages of ticks when isotopic values of ticks were compared with a panel of candidate vertebrate species. There was a positive shift in both δ(13)C and δ(15)N in adult A. americanum until 34 wk postnymphal bloodmeal. Through analyzing the isotopic signatures of eight potential vertebrate host species, we determined that the magnitude of this isotopic shift that occurred with tick age was minor compared with the heterogeneity in the δ(15)N and δ(13)C signatures among species. These results suggest that stable isotopes are a useful tool for understanding tick-host interactions., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The predominant human-biting tick throughout the southeastern United States is Amblyomma americanum. Its ability to transmit pathogens causing Lyme disease-like illnesses is a subject of ongoing controversy. Results of previous testing by the Department of Defense Human Tick Test Kit Program and other laboratories indicated that it is highly unlikely that A. americanum transmits any pathogen that causes Lyme disease. In contrast, a recent publication by Clark and colleagues (K. L. Clark, B. Leydet, and S. Hartman, Int. J. Med. Sci. 10:915-931, 2013) reported detection of Lyme group Borrelia in A. americanum using a nested-flagellin-gene PCR. We evaluated this assay by using it and other assays to test 1,097 A. americanum ticks collected from humans. Using the Clark assay, in most samples we observed nonspecific amplification and nonrepeatability of results on subsequent testing of samples. Lack of reaction specificity and repeatability is consistent with mispriming, likely due to high primer concentrations and low annealing temperatures in this protocol. In six suspect-positive samples, Borrelia lonestari was identified by sequencing of an independent gene region; this is not a Lyme group spirochete and is not considered zoonotic. B. burgdorferi was weakly amplified from one pool using some assays, but not others, and attempts to sequence the amplicon of this pool failed, as did attempts to amplify and sequence B. burgdorferi from the five individual samples comprising this pool. Therefore, B. burgdorferi was not confirmed in any sample. Our results do not support the hypothesis that A. americanum ticks are a vector for Lyme group Borrelia infections., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

In pines, self-pollination rates can be as high as 34% yet only 5% of viable seed is a product of self-fertilization. This decline in selfed seed viability is the consequence of post-fertilization exclusion mechanisms operating via the embryo lethal system. Recent molecular marker dissection studies suggest that the embryo lethal system is composed of semilethal factors dispersed across the genome, but it is not clear whether overdominant lethal factors are rare or representative. The study objective was to determine if overdominance was rare for the embryo lethal system in conifers. Three cohorts of selfed offspring from a single Pinus taeda parent were genotyped for nuclear microsatellites. Maximum likelihood tests based on distorted segregation ratios for single markers and for interval mapping were used to infer the degree of dominance. Four hypotheses about overdominance lethal factors were tested: (1) overdominant lethal factors rarely occur within the embryo lethal system, (2) overdominant lethal factors are rarely detected because they are transient and display stage-specific expression, (3) overdominant lethal factors are rarely detected due to tight linkage with rare marker alleles and (4) dominance estimation is unbiased by gametic selection. Four out of the seven chromosomal segments were linked to an overdominant lethal factor. One of these four segments had symmetric overdominance, an effect which persisted from embryo maturity through germination. Four overdominant lethal factors were linked to common and rare marker alleles. Gametic selection was not a source of bias in dominance estimation. Overdominant or pseudo-overdominant lethal factors are a common component of the conifer embryo lethal system.

Developing microsatellites from the large, highly duplicated conifer genome requires special tools. To improve the efficiency of developing Pinus taeda L. microsatellites, undermethylated (UM) DNA fragments were used to construct a microsatellite-enriched copy library. A methylation-sensitive restriction enzyme, McrBC, was used to enrich for UM DNA before library construction. Digested DNA fragments larger than 9 kb were then excised and digested with RsaI and used to construct nine dinucleotide and trinucleotide libraries. A total of 1016 microsatellite-positive clones were detected among 11 904 clones and 620 of these were unique. Of 245 primer sets that produced a PCR product, 113 could be developed as UM microsatellite markers and 70 were polymorphic. Inheritance and marker informativeness were tested for a random sample of 36 polymorphic markers using a three-generation outbred pedigree. Thirty-one microsatellites (86%) had single-locus inheritance despite the highly duplicated nature of the P. taeda genome. Nineteen UM microsatellites had highly informative intercross mating type configurations. Allele number and frequency were estimated for eleven UM microsatellites using a population survey. Allele numbers for these UM microsatellites ranged from 3 to 12 with an average of 5.7 alleles/locus. Frequencies for the 63 alleles were mostly in the low-common range; only 14 of the 63 were in the rare allele (q < 0.05) class. Enriching for UM DNA was an efficient method for developing polymorphic microsatellites from a large plant genome.