Dog ectoparasites as sentinels for pathogenic Rickettsia and Bartonella in rural Guatemala.

Background: Fleas and ticks serve as vectors of multiple pathogens in the genera Rickettsia and Bartonellathat cause diseases in humans and other animals. Although human rickettsiosis and bartonellosis have been reported in all countries in Central America, limited research has been conducted to investigate the natural cycles of flea- and tick-borne rickettsiosis and bartonellosis, especially in Guatemala. Methods: We evaluated dog parasites as sentinels for zoonotic disease risk in rural Guatemala by sampling ticks and fleas from dogs, which were then identified and individually screened for Rickettsia and Bartonella. Results: A total of 77 households were surveyed and 80.52% of them had dogs. Overall, 133 dogs were examined for fleas and ticks, of which 68.42% had fleas and 35.34% had ticks. A total of 433 fleas and 181 ticks were collected from the infested dogs, with an additional 33 ticks collected from house walls. Three flea species were identified: Ctenocephalides felis (70%), Echidnophaga gallinacea(11.8%), and Pulex sp. (17.8%). Among the collected ticks, 97% were Rhipicephalus sanguineus with the rest being Amyblyomma cajennense, A. auricularium, and A. ovale. Rickettsia felis were detected in six C. felis, in one Pulex sp., and in two R. sanguineus, while Candidatus R. senegalensis was detected in one C. felis. Bartonella was detected only in fleas, including three Pulexsp. infected with B. vinsonii subsp. Berkhoffii, B. henselae, and Bartonella sp., respectively, and 11 C. felis infected with B. henselae. Conclusions: This study reports Candidatus R. senegalensis and B. vinsonii subsp. Berkhoffiiin Guatemala for the first time, and indicates the potential risk of human and dog exposure to Rickettsia and Bartonella species. These results show that dogs provide critical information relevant to managing human potential exposure to flea- and tick-borne pathogens in rural Guatemala.

Domestic Dog Infection with Trypanosoma cruzi from Northern and Southern Regions of Mexico.

Background: Chagas disease or American trypanosomiasis, caused by Trypanosoma cruzi and vectored by triatomines, affects millions of people worldwide. In endemic countries including Mexico, infections in domestic animals, such as dogs, may affect the risk of human disease when they serve as a source of infection to vectors that subsequently infect humans. Materials and Methods: We conducted a cross-sectional study of 296 dogs from two cities near the northern and southern borders of Mexico: Reynosa, Tamaulipas, and Tuxtla Gutierrez, Chiapas. Infection was measured based on testing of blood using T. cruzi quantitative PCR (qPCR) and up to three antibody detection assays. The StatPak immunochromatographic assay was used to screen samples and the indirect fluorescent antibody (IFA) and multiplex microsphere immunoassay (MIA) tests were used as secondary tests on all samples that screened positive and a subset of negatives. Serologic positivity was defined based on reactivity on at least two independent tests. Results: Of the 280 samples tested for parasite DNA, two (0.7%) were positive, one of which (0.4%) was confirmed as T. cruzi discrete typing unit TcIV. Overall, 72 (24.3%) samples were reactive for T. cruzi antibodies via StatPak of which 8 were also positive using MIA and 2 were also positive using IFA (including one of the PCR-positive dogs). Overall, nine dogs (3.4%) met study criteria of positivity based on either/both serology or PCR tests. Positive dogs were found in both regions of Mexico; five (2.7%) from Reynosa and four (3.6%) from Tuxtla Gutierrez. We found no association between infection status and state of origin, sex, age group, breed group, neighborhood, and whether other pets lived in the home. Conclusion: Our results re-emphasize dogs' utility as sentinels for T. cruzi in Mexico and underscore the need for improved veterinary diagnostic tests and parasite surveillance at the household level in endemic countries.

Apparent absence of Trypanosoma cruzi in Mexican free-tailed bats (Tadarida brasiliensis) from Texas, USA.

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.

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

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

Fluralaner systemic treatment of chickens results in mortality in Triatoma gerstaeckeri, vector of the agent of Chagas disease

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.

Domestic Triatoma spp. Infections with Trypanosoma cruzi, Household Infestations, and Molecular Identification in Oaxaca, México.

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.

Brown Dog Tick (Rhipicephalus sanguineus Sensu Lato) Infection with Endosymbiont and Human Pathogenic Rickettsia spp., in Northeastern México.

Of the documented tick-borne diseases infecting humans in México, Rocky Mountain spotted fever (RMSF), caused by the Gram-negative bacterium Rickettsia rickettsii, is responsible for most fatalities. Given recent evidence of brown dog tick, Rhipicephalus sanguineus s.l., as an emerging vector of human RMSF, we aimed to evaluate dogs and their ticks for rickettsiae infections as an initial step in assessing the establishment of this pathosystem in a poorly studied region of northeastern México while evaluating the use of dogs as sentinels for transmission/human disease risk. We sampled owned dogs living in six disadvantaged neighborhoods of Reynosa, northeastern México to collect whole blood and ticks. Of 168 dogs assessed, tick infestation prevalence was 53%, composed of exclusively Rh. sanguineus s. l. (n = 2170 ticks). Using PCR and sequencing, we identified an overall rickettsiae infection prevalence of 4.1% (n = 12/292) in ticks, in which eight dogs harbored at least one infected tick. Rickettsiae infections included Rickettsia amblyommatis and Rickettsia parkeri, both of which are emerging human pathogens, as well as Candidatus Rickettsia andeanae. This is the first documentation of pathogenic Rickettsia species in Rh. sanguineus s.l. collected from dogs from northeastern México. Domestic dog infestation with Rickettsia-infected ticks indicates ongoing transmission; thus, humans are at risk for exposure, and this underscores the importance of public and veterinary health surveillance for these pathogens.

Domestic Dogs as Sentinels for West Nile Virus but not Aedes-borne Flaviviruses, Mexico.

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.

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

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

Effectiveness of fluralaner treatment regimens for the control of canine Chagas disease: A mathematical modeling study

Canine Chagas disease is caused by the protozoan parasite Trypanosoma cruzi and transmitted by insect triatomine vectors known as kissing bugs. The agent can cause cardiac damage and long-term heart disease and death in humans, dogs, and other mammals. In laboratory settings, treatment of dogs with systemic insecticides has been shown to be highly efficacious at killing triatomines that feed on treated dogs. Method We developed compartmental vector-host models of T. cruzi transmission between the triatomine and dog population accounting for the impact of seasonality and triatomine migration on disease transmission dynamics. We considered a single vector-host model without seasonality, and model with seasonality, and a spatially coupled model. We used the models to evaluate the effectiveness of the insecticide fluralaner with different durations of treatment regimens for reducing T. cruzi infection in different transmission settings. Results In low and medium transmission settings, our model showed a marginal difference between the 3-month and 6-month regimens for reducing T. cruzi infection among dogs. The difference increases in the presence of seasonality and triatomine migration from a sylvatic transmission setting. In high transmission settings, the 3-month regimen was substantially more effective in reducing T. cruzi infections in dogs than the other regimens. Our model showed that increased migration rate reduces fluralaner effectiveness in all treatment regimens, but the relative reduction in effectiveness is minimal during the first years of treatment. However, if an additional 10% or more of triatomines killed by dog treatment were eaten by dogs, treatment could increase T. cruzi infections in the dog population at least during the first year of treatment. Conclusion Our analysis shows that treating all peridomestic dogs every three to six months for at least five years could be an effective measure to reduce T. cruzi infections in dogs and triatomines in peridomestic transmission settings. However, further studies at the local scale are needed to better understand the potential impact of routine use of fluralaner treatment on increasing dogs' consumption of dead triatomines.

Vector-Borne Blood Parasites of the Great-Tailed Grackle (Quiscalus mexicanus) in East-Central Texas, USA.

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.

Selected cardiac abnormalities in Trypanosoma cruzi serologically positive, discordant, and negative working dogs along the Texas-Mexico border.

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.

Rapid detection of human blood in triatomines (kissing bugs) utilizing a lateral flow immunochromatographic assay - A pilot study.

OBJECTIVES: We tested a rapid and specific immunochromatographic assay (that detects human blood in forensic samples) to determine if human blood was present in triatomines and their fecal excreta. METHODS: We fed Triatoma rubida human blood (positive control) or mouse blood (negative control) and performed the assay on the abdominal contents and fecal excreta. Triatomine field specimens collected in and around human habitations and excreta were also tested. FINDINGS: The assay was positive in triatomines fed human blood (N = 5/5) and fecal excreta from bugs known to have ingested human blood (N = 5/5). Bugs feeding on mice (N = 15/15) and their fecal excreta (N = 8/8) were negative for human blood. Human blood was detected in 47% (N = 23/49) triatomines, representing six different species, collected in the field. MAIN CONCLUSIONS: The pilot study shows that this rapid and specific test may have applications in triatomine research. Further study is needed to determine the sensitivity of this assay compared to other well-established techniques, such as DNA- and proteomics-based methodologies and the assay's application in the field.

Rapid detection of human blood in triatomines (kissing bugs) utilizing a lateral flow immunochromatographic assay - A pilot study

BACKGROUND DNA- and proteomics-based techniques are currently used to identify a triatomine human blood meal. These methods are time consuming, require access to laboratories with sophisticated equipment, and trained personnel. OBJECTIVES We tested a rapid and specific immunochromatographic assay (that detects human blood in forensic samples) to determine if human blood was present in triatomines and their fecal excreta. METHODS We fed Triatoma rubida human blood (positive control) or mouse blood (negative control) and performed the assay on the abdominal contents and fecal excreta. Triatomine field specimens collected in and around human habitations and excreta were also tested. FINDINGS The assay was positive in triatomines fed human blood (N = 5/5) and fecal excreta from bugs known to have ingested human blood (N = 5/5). Bugs feeding on mice (N = 15/15) and their fecal excreta (N = 8/8) were negative for human blood. Human blood was detected in 47% (N = 23/49) triatomines, representing six different species, collected in the field. MAIN CONCLUSIONS The pilot study shows that this rapid and specific test may have applications in triatomine research. Further study is needed to determine the sensitivity of this assay compared to other well-established techniques, such as DNA- and proteomics-based methodologies and the assay's application in the field.

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

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

Widespread Trypanosoma cruzi infection in government working dogs along the Texas-Mexico border: Discordant serology, parasite genotyping and associated vectors.

BACKGROUND: Chagas disease, caused by the vector-borne protozoan Trypanosoma cruzi, is increasingly recognized in the southern U.S. Government-owned working dogs along the Texas-Mexico border could be at heightened risk due to prolonged exposure outdoors in habitats with high densities of vectors. We quantified working dog exposure to T. cruzi, characterized parasite strains, and analyzed associated triatomine vectors along the Texas-Mexico border. METHODOLOGY/PRINCIPLE FINDINGS: In 2015-2016, we sampled government working dogs in five management areas plus a training center in Texas and collected triatomine vectors from canine environments. Canine serum was tested for anti-T. cruzi antibodies with up to three serological tests including two immunochromatographic assays (Stat-Pak and Trypanosoma Detect) and indirect fluorescent antibody (IFA) test. The buffy coat fraction of blood and vector hindguts were tested for T. cruzi DNA and parasite discrete typing unit was determined. Overall seroprevalence was 7.4 and 18.9% (n = 528) in a conservative versus inclusive analysis, respectively, based on classifying weakly reactive samples as negative versus positive. Canines in two western management areas had 2.6-2.8 (95% CI: 1.0-6.8 p = 0.02-0.04) times greater odds of seropositivity compared to the training center. Parasite DNA was detected in three dogs (0.6%), including TcI and TcI/TcIV mix. Nine of 20 (45%) T. gerstaeckeri and T. rubida were infected with TcI and TcIV; insects analyzed for bloodmeals (n = 11) fed primarily on canine (54.5%). CONCLUSIONS/SIGNIFICANCE: Government working dogs have widespread exposure to T. cruzi across the Texas-Mexico border. Interpretation of sample serostatus was challenged by discordant results across testing platforms and very faint serological bands. In the absence of gold standard methodologies, epidemiological studies will benefit from presenting a range of results based on different tests/interpretation criteria to encompass uncertainty. Working dogs are highly trained in security functions and potential loss of duty from the clinical outcomes of infection could affect the work force and have broad consequences.

Salmonella Surveillance Among Great-Tailed Grackles (Quiscalus mexicanus) and Other Urban Bird Species in Eastern Texas.

Wild birds may play an important role in maintaining and transmitting Salmonella. Their ability to travel large distances and their proximity to human habitations could make them a vehicle for bridging Salmonella from wild and domestic animals to humans. To determine the potential public health risk presented by urban birds, we investigated the prevalence of Salmonella among great-tailed grackles (Quiscalus mexicanus) and other cohabiting urban bird species. Fecal samples were collected from 114 birds communally roosting in parking lots of retail locations in Brazos County, Texas, from February through July of 2015. Great-tailed grackles and European starlings (Sturnus vulgaris) were the predominant species sampled. Standard bacteriologic culture methods were used to isolate Salmonella from samples, and isolates were characterized by serotyping and antimicrobial susceptibility testing. Overall, 1.8% (2/114) of samples were confirmed positive for Salmonella. Both positive birds were great-tailed grackles sampled in June, yielding a 2.6% (2/76) apparent prevalence among this species. Isolates were serotyped as Salmonella Typhimurium and found to be pan-susceptible based on the National Antimicrobial Resistance Monitoring System (NARMS) panel of antimicrobial agents. The occurrence of Salmonella in great-tailed grackles represents a potential threat to public health, particularly considering their population size and tendency to congregate near human establishments such as grocery stores.