Citizen Science Provides an Efficient Method for Broad-Scale Tick-Borne Pathogen Surveillance of Ixodes pacificus and Ixodes scapularis across the United States.

Tick-borne diseases have expanded over the last 2 decades as a result of shifts in tick and pathogen distributions. These shifts have significantly increased the need for accurate portrayal of real-time pathogen distributions and prevalence in hopes of stemming increases in human morbidity. Traditionally, pathogen distribution and prevalence have been monitored through case reports or scientific collections of ticks or reservoir hosts, both of which have challenges that impact the extent, availability, and accuracy of these data. Citizen science tick collections and testing campaigns supplement these data and provide timely estimates of pathogen prevalence and distributions to help characterize and understand tick-borne disease threats to communities. We utilized our national citizen science tick collection and testing program to describe the distribution and prevalence of four Ixodes-borne pathogens, Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia microti, across the continental United States. IMPORTANCE In the 21st century, zoonotic pathogens continue to emerge, while previously discovered pathogens continue to have changes within their distribution and prevalence. Monitoring these pathogens is resource intensive, requiring both field and laboratory support; thus, data sets are often limited within their spatial and temporal extents. Citizen science collections provide a method to harness the general public to collect samples, enabling real-time monitoring of pathogen distribution and prevalence.

Borreliosis Transmission from Ticks Associated with Desert Tortoise Burrows: Examples of Tick-Borne Relapsing Fever in the Mojave Desert.

Ticks transmit pathogens and parasitize wildlife in turn causing zoonotic diseases in many ecosystems. Argasid ticks, such as Ornithodoros spp., harbor and transmit Borrelia spp., resulting in tick-borne relapsing fever (TBRF) in people. In the western United States, TBRF is typically associated with the bite of an infected Ornithodoros hermsi tick found in habitats at high elevations (>1500 ft). This report describes the first TBRF cases in people in the Mojave Desert (Clark County, NV). Individuals documented in these case studies were exposed to Ornithodoros ticks during excavation of soil burrows associated with Mojave Desert tortoises (Gopherus agassizii), with bacteria from one of the human case's blood sample genetically matching to Borrelia turicatae as determined by quantitative PCR and sequencing. Our findings should serve as a precaution to individuals working with tortoises or animal burrows, or those in contact with Ornithodoros ticks in this region.

Examining Prevalence and Diversity of Tick-Borne Pathogens in Questing Ixodes pacificus Ticks in California.

Tick-borne diseases in California include Lyme disease (caused by Borrelia burgdorferi), infections with Borrelia miyamotoi, and human granulocytic anaplasmosis (caused by Anaplasma phagocytophilum). We surveyed multiple sites and habitats (woodland, grassland, and coastal chaparral) in California to describe spatial patterns of tick-borne pathogen prevalence in western black-legged ticks (Ixodes pacificus). We found that several species of Borrelia-B. burgdorferi, Borrelia americana, and Borrelia bissettiae-were observed in habitats, such as coastal chaparral, that do not harbor obvious reservoir host candidates. Describing tick-borne pathogen prevalence is strongly influenced by the scale of surveillance: aggregating data from individual sites to match jurisdictional boundaries (e.g., county or state) can lower the reported infection prevalence. Considering multiple pathogen species in the same habitat allows a more cohesive interpretation of local pathogen occurrence. IMPORTANCE Understanding the local host ecology and prevalence of zoonotic diseases is vital for public health. Using tick-borne diseases in California, we show that there is often a bias to our understanding and that studies tend to focus on particular habitats, e.g., Lyme disease in oak woodlands. Other habitats may harbor a surprising diversity of tick-borne pathogens but have been neglected, e.g., coastal chaparral. Explaining pathogen prevalence requires descriptions of data on a local scale; otherwise, aggregating the data can misrepresent the local dynamics of tick-borne diseases.

Equids engineer desert water availability.

Megafauna play important roles in the biosphere, yet little is known about how they shape dryland ecosystems. We report on an overlooked form of ecosystem engineering by donkeys and horses. In the deserts of North America, digging of ≤2-meter wells to groundwater by feral equids increased the density of water features, reduced distances between waters, and, at times, provided the only water present. Vertebrate richness and activity were higher at equid wells than at adjacent dry sites, and, by mimicking flood disturbance, equid wells became nurseries for riparian trees. Our results suggest that equids, even those that are introduced or feral, are able to buffer water availability, which may increase resilience to ongoing human-caused aridification.

Predicting the current and future distribution of the western black-legged tick, Ixodes pacificus, across the Western US using citizen science collections.

In the twenty-first century, ticks and tick-borne diseases have expanded their ranges and impact across the US. With this spread, it has become vital to monitor vector and disease distributions, as these shifts have public health implications. Typically, tick-borne disease surveillance (e.g., Lyme disease) is passive and relies on case reports, while disease risk is calculated using active surveillance, where researchers collect ticks from the environment. Case reports provide the basis for estimating the number of cases; however, they provide minimal information on vector population or pathogen dynamics. Active surveillance monitors ticks and sylvatic pathogens at local scales, but it is resource-intensive. As a result, data are often sparse and aggregated across time and space to increase statistical power to model or identify range changes. Engaging public participation in surveillance efforts allows spatially and temporally diverse samples to be collected with minimal effort. These citizen-driven tick collections have the potential to provide a powerful tool for tracking vector and pathogen changes. We used MaxEnt species distribution models to predict the current and future distribution of Ixodes pacificus across the Western US through the use of a nationwide citizen science tick collection program. Here, we present niche models produced through citizen science tick collections over two years. Despite obvious limitations with citizen science collections, the models are consistent with previously-predicted species ranges in California that utilized more than thirty years of traditional surveillance data. Additionally, citizen science allows for an expanded understanding of I. pacificus distribution in Oregon and Washington. With the potential for rapid environmental changes instigated by a burgeoning human population and rapid climate change, the development of tools, concepts, and methodologies that provide rapid, current, and accurate assessment of important ecological qualities will be invaluable for monitoring and predicting disease across time and space.

Leptospira in river and soil in a highly endemic area of Ecuador.

BACKGROUND: Leptospira are shed into the environment via urine of infected animals. Rivers are thought to be an important risk factor for transmission to humans, though much is unknown about the types of environment or characteristics that favor survival. To address this, we screened for Leptospira DNA in two rivers in rural Ecuador where Leptospirosis is endemic. RESULTS: We collected 112 longitudinal samples and recorded pH, temperature, river depth, precipitation, and dissolved oxygen. We also performed a series of three experiments designed to provide insight into Leptospira presence in the soil. In the first soil experiment, we characterized prevalence and co-occurrence of Leptospira with other bacterial taxa in the soil at dispersed sites along the rivers (n = 64). In the second soil experiment, we collected 24 river samples and 48 soil samples at three points along eight transects to compare the likelihood of finding Leptospira in the river and on the shore at different distances from the river. In a third experiment, we tested whether Leptospira presence is associated with soil moisture by collecting 25 soil samples from two different sites. In our river experiment, we found pathogenic Leptospira in only 4 (3.7%) of samples. In contrast, pathogenic Leptospira species were found in 22% of shore soil at dispersed sites, 16.7% of soil samples (compared to 4.2% of river samples) in the transects, and 40% of soil samples to test for associations with soil moisture. CONCLUSIONS: Our data are limited to two sites in a highly endemic area, but the scarcity of Leptospira DNA in the river is not consistent with the widespread contention of the importance of river water for leptospirosis transmission. While Leptospira may be shed directly into the river, onto the shores, or washed into the river from more remote sites, massive dilution and limited persistence in rivers may reduce the environmental load and therefore, the epidemiological significance of such sources. It is also possible that transmission may occur more frequently on shores where people are liable to be barefoot. Molecular studies that further explore the role of rivers and water bodies in the epidemiology of leptospirosis are needed.

Integration of choline geranate into electrospun protein scaffolds affords antimicrobial activity to biomaterials used for cutaneous wound healing.

Wound healing attempts to maintain homeostasis in the wound while minimizing the risk of infection to the tissue by foreign agents, such as opportunistic bacterial pathogens. Biofilms established by these pathogens are a common cause of chronic infections that slow the healing process. Preparation of skin wound healing devices comprised of electrospun proteins associated with skin have been shown to accelerate the healing process relative to conventional wound dressings. In this work, we have developed electrospinning methods to incorporate the antimicrobial ionic liquid/deep eutectic solvent choline geranate (CAGE) into these devices. Integration of CAGE into the dressing material was verified via 1 H nuclear magnetic resonance spectrometry, and the effect on the material property of the resultant devices were assessed using scanning electron microscopy. CAGE-containing devices demonstrate a concentration-dependent inactivation of exogenously applied solutions of both gram-positive and gram-negative pathogens (Enterococcus sp and Pseudomonas aeruginosa, respectively), but maintain their ability to serve as a compatible platform for proliferation of human dermal neonatal fibroblasts.

Isolation and Molecular Characterization of Tick-Borne Relapsing Fever Borrelia Infecting Ornithodoros (Pavlovskyella) verrucosus Ticks Collected in Ukraine.

BACKGROUND: Tick-borne relapsing fever (TBRF) is a neglected zoonotic bacterial disease known to occur on 5 continents. We report a laboratory-acquired case of TBRF caused by Borrelia caucasica, which is endemic in Ukraine and transmitted by Ornithodoros verrucosus ticks. METHODS: We isolated spirochetes and characterized them by partially sequencing the 16s ribosomal ribonucleic acid (rrs), flagellin (flaB), and deoxyribonucleic acid gyrase (gyrB) genes and conducting a phylogenetic analysis. RESULTS: These analyses revealed a close relationship of Ukrainian spirochetes with the Asian TBRF species, Borrelia persica. The taxonomic and nomenclature problems related to insufficient knowledge on the spirochetes and their vectors in the region are discussed. CONCLUSIONS: Although these findings enhance our understanding of species identities for TBRF Borrelia in Eurasia, further work is required to address the neglected status of TBRF in this part of the world. Public health practitioners should consider TBRF and include the disease into differential diagnosis of febrile illnesses with unknown etiology.

COYOTES (CANIS LATRANS) IN ARIZONA, USA, EXHIBIT IMMUNE AND GENETIC EVIDENCE OF RICKETTSIAL INFECTIONS.

Rocky Mountain spotted fever (RMSF), caused by the bacterium Rickettsia rickettsii, was recognized as endemic in Arizona, US after a 2002 outbreak and has since been a public health concern. The brown dog tick (Rhipicephalus sanguineus sensu lato) is the principal vector of this pathogen in Arizona. Domesticated dogs (Canis lupus familiaris) are the tick's main host, so free-roaming dogs in peridomestic areas have been named the primary risk factor for human cases of RMSF. However, the sudden emergence and long-distance dispersal of the pathogen have not been adequately explained, and one possible mechanism could include wildlife. Coyotes (Canis latrans) are wide ranging in Arizona and closely related to dogs, so it is possible that brown dog ticks parasitize coyotes and infect them. Although R. rickettsii is the most severe spotted fever group (SFG) rickettsial pathogen in humans, others occur in Arizona, and antibodies raised against them are cross-reactive, so we more-broadly hypothesized that coyotes in Arizona are exposed to SFG rickettsiae. We collected coyote tissues in spring 2016 and 2017. We tested sera for antibodies to R. rickettsii and found 9% (8/94) of samples were antibody-positive with titers of ≥256. Subsequent quantitative PCR analyses of skin showed evidence for Rickettsia spp. in 2.9% (4/138) of samples. These data suggest that coyotes have a role in the maintenance of SFG rickettsiae in Arizona. Further investigation is warranted to reveal which specific pathogen-vector complexes act on coyotes in the region and whether they represent a risk to human health.

Time of year and outdoor recreation affect human exposure to ticks in California, United States.

Interactions between humans and ticks are often measured indirectly, using surveillance of tick population abundance and pathogen prevalence, or reported human disease data. We used data garnered as part of a free national citizen science research effort to describe actual human exposures to ticks in California. Human-biting ticks (n = 1,905) submitted for identification were predominantly western black-legged ticks (Ixodes pacificus) (68%), American dog ticks (Dermacentor variabilis) (24%), and Pacific Coast ticks (Dermacentor occidentalis) (7%). Tick exposure occurred predominantly during recreational use of the outdoors, rather than exposure near the home environment. Tick submissions peaked in May, but human exposure to ticks occurred throughout the year. Adult I. pacificus were most frequently found on humans during March-May, though previous research demonstrates that questing adults on vegetation are more abundant earlier in the winter.

Citizen science informs human-tick exposure in the Northeastern United States.

BACKGROUND: Tick-borne disease is the result of spillover of pathogens into the human population. Traditionally, literature has focused on characterization of tick-borne disease pathogens and ticks in their sylvatic cycles. A limited amount of research has focused on human-tick exposure in this system, especially in the Northeastern United States. Human-tick interactions are crucial to consider when assessing the risk of tick-borne disease since a tick bite is required for spillover to occur. METHODS: Citizen scientists collected ticks from the Northeastern US through a free nationwide program. Submitted ticks were identified to species, stage, and sex. Blacklegged ticks, Ixodes scapularis, were tested for the presence of Borrelia burgdorferi sensu lato (s.l.) and hard-tick relapsing fever Borrelia. Seasonality of exposure and the citizen science activity during tick exposure was recorded by the citizen scientist. A negative binomial model was fit to predict county level CDC Lyme disease cases in 2016 using citizen science Ixodes scapularis submissions, state, and county population as predictor variables. RESULTS: A total of 3740 submissions, comprising 4261 ticks, were submitted from the Northeastern US and were reported to be parasitizing humans. Of the three species submitted, blacklegged ticks were the most prevalent followed by American dog ticks and lone star ticks. Submissions peaked in May with the majority of exposure occurring during every-day activities. The most common pathogen in blacklegged ticks was B. burgdorferi s.l. followed by hard-tick relapsing fever Borrelia. Negative binomial model performance was best in New England states followed by Middle Atlantic states. CONCLUSIONS: Citizen science provides a low-cost and effective methodology for describing the seasonality and characteristics of human-tick exposure. In the Northeastern US, everyday activities were identified as a major mechanism for tick exposure, supporting the role of peri-domestic exposure in tick-borne disease. Citizen science provides a method for broad pathogen and tick surveillance, which is highly related to human disease, allowing for inferences to be made about the epidemiology of tick-borne disease.

Borrelia miyamotoi Infections in Small Mammals, California, USA.

Surveillance to investigate the wildlife-vector transmission cycle of the human pathogen Borrelia miyamotoi in California, USA, revealed infections in dusky-footed woodrats, brush mice, and California mice. Phylogenetic analyses suggest a single, well-supported clade of B. miyamotoi is circulating in California.

Serologic assessment for exposure to spotted fever group rickettsiae in dogs in the Arizona-Sonora border region.

Rocky Mountain spotted fever (RMSF) is a severe tick-borne rickettsial illness. In the south-western United States and Mexico, RMSF displays unique epidemiologic and ecologic characteristics, including Rhipicephalus sanguineus sensu lato (brown dog tick) as the primary vector. Expansion and spread of the disease from hyperendemic regions of Arizona or Mexico to new areas is a key public health concern. Dogs are thought to play an important role in the emergence and circulation of R. rickettsii in these regions and are often one of earliest indicators of RMSF presence. A canine serosurvey was conducted in 2015 among owned and stray dogs at rabies clinic and animal shelters in three southern Arizona counties where RMSF had not previously been identified. Of the 217 dogs sampled, 11 (5.1%) tested positive for spotted fever group rickettsia (SFGR) IgG antibodies, with seropositivity ranging from 2.9% to 12.2% across the three counties. Large dogs were significantly more likely than small dogs to have positive titres reactive with R. rickettsii; no additional statistically significant relationships were observed between seropositivity of canine age, sex, neuter or ownership status. In addition, 17 (7.8%) dogs had ticks attached at the time of sampling, and stray dogs were significantly more likely to have ticks present than owned dogs (p < 0.001). All 57 ticks collected were identified as Rh. sanguineus s.l., and four (7%) had DNA evidence of genera-wide Rickettsia species. The results of this project demonstrated canine seroprevalence levels lower than those previously reported from dogs in highly endemic areas, indicating a low risk of SFGR transmission to humans in the southern Arizona border region at this time. Continued surveillance is critical to identify SFGR emergence in new geographic regions and to inform prevention efforts for humans and dogs in those areas.

Comparative pan-genomic analyses of Orientia tsutsugamushi reveal an exceptional model of bacterial evolution driving genomic diversity.

Orientia tsutsugamushi, formerly Rickettsia tsutsugamushi, is an obligate intracellular pathogen that causes scrub typhus, an underdiagnosed acute febrile disease with high morbidity. Scrub typhus is transmitted by the larval stage (chigger) of Leptotrombidium mites and is irregularly distributed across endemic regions of Asia, Australia and islands of the western Pacific Ocean. Previous work to understand population genetics in O. tsutsugamushi has been based on sub-genomic sampling methods and whole-genome characterization of two genomes. In this study, we compared 40 genomes from geographically dispersed areas and confirmed patterns of extensive homologous recombination likely driven by transposons, conjugative elements and repetitive sequences. High rates of lateral gene transfer (LGT) among O. tsutsugamushi genomes appear to have effectively eliminated a detectable clonal frame, but not our ability to infer evolutionary relationships and phylogeographical clustering. Pan-genomic comparisons using 31 082 high-quality bacterial genomes from 253 species suggests that genomic duplication in O. tsutsugamushi is almost unparalleled. Unlike other highly recombinant species where the uptake of exogenous DNA largely drives genomic diversity, the pan-genome of O. tsutsugamushi is driven by duplication and divergence. Extensive gene innovation by duplication is most commonly attributed to plants and animals and, in contrast with LGT, is thought to be only a minor evolutionary mechanism for bacteria. The near unprecedented evolutionary characteristics of O. tsutsugamushi, coupled with extensive intra-specific LGT, expand our present understanding of rapid bacterial evolutionary adaptive mechanisms.

Using citizen science to describe the prevalence and distribution of tick bite and exposure to tick-borne diseases in the United States.

Tick-borne pathogens are increasing their range and incidence in North America as a consequence of numerous factors including improvements in diagnostics and diagnosis, range expansion of primary vectors, changes in human behavior, and an increasing understanding of the diversity of species of pathogens that cause human disease. Public health agencies have access to human incidence data on notifiable diseases e.g., Borrelia burgdorferi, the causative agent of Lyme disease, and often local pathogen prevalence in vector populations. However, data on exposure to vectors and pathogens can be difficult to determine e.g., if disease does not occur. We report on an investigation of exposure to ticks and tick-borne bacteria, conducted at a national scale, using citizen science participation. 16,080 ticks were submitted between January 2016 and August 2017, and screened for B. burgdorferi, B. miyamotoi, Anaplasma phagocytophilum, and Babesia microti. These data corroborate entomologic investigations of tick distributions in North America, but also identify patterns of local disease risk and tick contact with humans throughout the year in numerous species of ticks and associated pathogens.

Ecological niche modeling and distribution of Ornithodoros hermsi associated with tick-borne relapsing fever in western North America.

Tick-borne relapsing fever in western North America is a zoonosis caused by the spirochete bacterium, Borrelia hermsii, which is transmitted by the bite of infected Ornithodoros hermsi ticks. The pathogen is maintained in natural cycles involving small rodent hosts such as chipmunks and tree squirrels, as well as the tick vector. In order for these ticks to establish sustained and viable populations, a narrow set of environmental parameters must exist, primarily moderate temperatures and moderate to high amounts of precipitation. Maximum Entropy Species Distribution Modeling (Maxent) was used to predict the species distribution of O. hermsi and B. hermsii through time and space based on current climatic trends and future projected climate changes. From this modeling process, we found that the projected current distributions of both the tick and spirochete align with known endemic foci for the disease. Further, global climate models predict a shift in the distribution of suitable habitat for the tick vector to higher elevations. Our predictions are useful for targeting surveillance efforts in areas of high risk in western North America, increasing the efficiency and accuracy of public health investigations and vector control efforts.

Preliminary evidence that age and sex affect exercise-induced hTERT expression.

The ability to repair cellular damage is reduced with aging, resulting in cellular senescence. Telomeres shorten as cells divide but the rate of telomere attrition is modulated by telomerase, an enzyme that adds nucleotides to the chromosome. Shelterin is a protein complex that acts as a negative regulator of telomerase. The aim of the present study was to investigate age-related differences in telomerase and shelterin responses to acute exercise. We hypothesized that acute exercise would stimulate an increased activity of telomerase (measured by telomerase reverse transcriptase, hTERT) without an increase in activity of shelterin (measured by telomeric repeat binding factor 2, TRF2) in both young and older individuals and that hTERT response would be attenuated in older individuals. Young (22±2y, n=11) and older (60±2y, n=8) men and women performed 30min of cycling. Blood was collected pre-exercise and 30, 60, and 90-min post-exercise. The trial induced a significant hTERT response in the cohort as a whole (p<0.05) with greater increases in the young as compared to the older group (time-by-group interaction p<0.05). As expected, TRF2 did not change in response to the trial, however older individuals had a higher TRF2 response at 60min (p<0.05). There was an unexpected sex difference, regardless of age, where men had significantly greater hTERT and TRF2 responses to the acute exercise as compared to women (p<0.05). These data support the hypothesis that aging is associated with attenuated telomerase activation in response to high-intensity exercise; however, this was only evident in men.

Meta-analysis to estimate the load of Leptospira excreted in urine: beyond rats as important sources of transmission in low-income rural communities.

BACKGROUND: Leptospirosis is a major zoonotic disease with widespread distribution and a large impact on human health. Carrier animals excrete pathogenic Leptospira primarily in their urine. Infection occurs when the pathogen enters a host through mucosa or small skin abrasions. Humans and other animals are exposed to the pathogen by direct contact with urine, contaminated soil or water. While many factors influence environmental cycling and the transmission of Leptospira to humans, the load of pathogenic Leptospira in the environment is likely to play a major role. Peridomestic rats are often implicated as a potential source of human disease; however exposure to other animals is a risk factor as well. The aim of this report is to highlight the importance of various carrier animals in terms of the quantity of Leptospira shed into the environment. For this, we performed a systematic literature review and a meta-analysis of the amount of pathogen that various animal species shed in their urine. RESULTS: The quantity of pathogen has been reported for cows, deer, dogs, humans, mice, and rats, in a total of 14 research articles. We estimated the average Leptospira per unit volume shed by each animal species, and the daily environmental contribution by considering the total volume of urine excreted by each carrier animal. Rats excrete the highest quantity of Leptospira per millilitre of urine (median = 5.7 × 106 cells), but large mammals excrete much more urine and thus shed significantly more Leptospira per day (5.1 × 108 to 1.3 × 109 cells). CONCLUSIONS: Here we illustrate how, in a low-income rural Ecuadorian community, host population demographics, and prevalence of Leptospira infection can be integrated with estimates of shed Leptospira to suggest that peridomestic cattle may be more important than rats in environmental cycling and ultimately, transmission to humans.

COMPARISON OF CURRENT METHODS FOR THE DETECTION OF CHRONIC MYCOPLASMAL URTD IN WILD POPULATIONS OF THE MOJAVE DESERT TORTOISE (GOPHERUS AGASSIZII).

Pathogens that cause subclinical diseases or exhibit low infection intensities are difficult to quantify in wild populations. Mojave desert tortoises ( Gopherus agassizii ) have been the focus of much research aimed at measuring the presence of upper respiratory disease (URTD) and URTD-associated pathogens, and techniques used to quantify disease in Gopherus species have also been used for disease surveillance in other species of turtles and tortoises of conservation concern. Published surveys of G. agassizii populations have found a relatively low prevalence of URTD, with most URTD-positive animals exhibiting moderate, intermittent signs of morbidity. Therefore, multiple tests have been developed to quantify URTD including genetic detection of the pathogens Mycoplasma agassizii and Mycoplasma testudineum , detection of M. agassizii -specific antibodies, and standardized quantification of clinical signs of URTD and body condition. These diagnostic tests have only been compared in diseased or moribund, semicaptive animals. We compared diagnostic techniques (TaqMan® and SYBR™ Green qPCR, serology, and visible examination) to detect M. agassizii -associated URTD in 126 wild desert tortoises sampled in Nevada and California, US in 2010. All had healthy body condition indices and none exhibited more than mild-to-moderate visual signs of URTD. Pairwise comparisons of diagnostic techniques indicated poor performance in diagnosing disease in individual animals. We found stronger, but inconsistent, statistical associations among diagnostic techniques at the population level. Our findings have implications for quantifying subclinical respiratory disease in tortoises.