On-host flea phenology and flea-borne pathogen surveillance among mammalian wildlife of the pineywoods of East Texas.

Flea-borne diseases are endemic in Texas, U.S.A., with an increasing incidence of flea-borne typhus and cat scratch disease. Knowledge of flea natural history could provide information to protect public health, yet many knowledge gaps remain outside of plague-endemic regions. Our objective was to characterize seasonal activity patterns of fleas on common mammalian wildlife species and test fleas and wildlife for Rickettsia and Bartonella pathogens. We performed one year of monthly trapping for rodents and medium-sized mammals in a national forest with high recreational use and urban encroachment in East Texas. From 90 mammal captures representing seven species, 101 fleas were collected representing Polygenis spp., Ctenocephalides felis, and Orchopeas species. Virginia opossums (Didelphis virginianus) hosted 99% of the collected fleas (100 fleas) and a single flea was on an eastern woodrat (Neotoma floridana). Flea infestation prevalence of opossums was 79% (23/29). Mean flea abundance was 4.39 fleas, with intensity peaking in spring. One cat flea removed from an opossum was positive for Bartonella henselae. Furthermore, we identified tissue or blood of four raccoons (Procyon lotor) and one golden mouse (Ochrotomys nuttalli) positive for Rickettsia amblyommatis. These findings provide an ecological basis for the maintenance of vectors and pathogens from sylvatic settings.

Dirofilaria immitis and Dirofilaria striata (Spirurida: Onchocercidae) detected in wild carnivores from Texas, United States.

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

Macro-parasites and micro-parasites co-exist in rodent communities but are associated with different community-level parameters.

Wildlife species are often heavily parasitized by multiple infections simultaneously. Yet research on sylvatic transmission cycles, tend to focus on host interactions with a single parasite and neglects the influence of co-infections by other pathogens and parasites. Co-infections between macro-parasites and micro-parasites can alter mechanisms that regulate pathogenesis and are important for understanding disease emergence and dynamics. Wildlife rodent hosts in the Lyme disease system are infected with macro-parasites (i.e., ticks and helminths) and micro-parasites (i.e., Borrelia spp.), however, there has not been a study that investigates the interaction of all three parasites (i.e., I. pacificus, Borrelia spp., and helminths) and how these co-infections impact prevalence of micro-parasites. We live-trapped rodents in ten sites in northern California to collect feces, blood, ear tissue, and attached ticks. These samples were used to test for infection status of Borrelia species (i.e., micro-parasite), and describe the burden of ticks and helminths (i.e., macro-parasites). We found that some rodent hosts were co-infected with all three parasites, however, the burden or presence of concurrent macro-parasites were not associated with Borrelia infections. For macro-parasites, we found that tick burdens were positively associated with rodent Shannon diversity while negatively associated with predator diversity, whereas helminth burdens were not significantly associated with any host community metric. Ticks and tick-borne pathogens are associated with rodent host diversity, predator diversity, and abiotic factors. However, it is still unknown what factors helminths are associated with on the community level. Understanding the mechanisms that influence co-infections of multiple types of parasites within and across hosts is an increasingly critical component of characterizing zoonotic disease transmission and maintenance.

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.

Host infection and community composition predict vector burden.

Lyme disease is the most prevalent vector-borne disease in the United States, yet critical gaps remain in our understanding of tick and host interactions that shape disease dynamics. Rodents such as deer mice (Peromyscus spp.) and dusky-footed woodrats (Neotoma fuscipes) are key reservoirs for Borrelia burgdorferi, the etiological bacterium of Lyme disease, and can vary greatly in abundance between habitats. The aggregation of Ixodes pacificus, the western black-legged tick, on rodent hosts is often assumed to be constant across various habitats and not dependent on the rodent or predator communities; however, this is rarely tested. The factors that determine tick burdens on key reservoir hosts are important in estimating Lyme disease risk because larger tick burdens can amplify pathogen transmission. This study is the first to empirically measure I. pacificus larval burdens on competent reservoir hosts as a function of community factors such as rodent diversity, predator diversity, and questing tick abundance. Rodents were live trapped at oak woodland sites to collect tick burdens and tissue samples to test for infection with Borrelia burgdorferi sensu lato. We found that N. fuscipes tick burdens were negatively correlated with predator diversity, but positively correlated with questing I. pacificus larvae. In addition, rodent hosts that were infected with B. burgdorferi sensu lato tend to have higher burdens of larval ticks. These results demonstrate that tick burdens can be shaped by variability between individuals, species, and the broader host community with consequences for transmission and prevalence of tick-borne pathogens.

A Beginner's Guide to Collecting Questing Hard Ticks (Acari: Ixodidae): A Standardized Tick Dragging Protocol.

Tick-borne diseases are emerging globally, necessitating increased research and coordination of tick surveillance practices. The most widely used technique for active collection of host-seeking, human-biting tick vectors is 'tick dragging', by which a cloth is dragged across the top of the vegetation or forest floor and regularly checked for the presence of ticks. Use of variable dragging protocols limits the ability of researchers to combine data sets for comparative analyses or determine patterns and trends across different spatial and temporal scales. Standardization of tick drag collection and reporting methodology will greatly benefit the field of tick-pathogen studies. Based on the recommendations of the Center for Disease Control and Prevention and other ecological considerations, we propose that tick dragging should be conducted to sample at least 750 m2 along linear transects when habitat allows in a manner that reduces bias in the sampled area, and report density of each tick species and life stage separately. A protocol for constructing a standard drag cloth, establishing linear transects, and drag sampling is presented, along with a downloadable datasheet that can be modified to suit the needs of different projects. Efforts to align tick surveillance according to these standard best practices will help generate robust data on tick population biology.