An ecological and epidemiological single-season survey of Anaplasma and Ehrlichia positive ticks in Victoria Falls National Park, Zimbabwe.

There is an urgent need for continued research on the ecology of tick-borne diseases in Africa. Our objective was to provide a preliminary description of the ecology and epidemiology of tick species, tick-borne pathogens, and animal hosts in Zimbabwe, focusing efforts at Victoria Falls National Park, for a single season. We tested the hypothesis that tick surveillance and pathogen screening data can be used to model associations among ticks, hosts, and pathogens. We collected ticks from domesticated animals and wildlife in Zimbabwe and screened the ticks for the presence of Anaplasma and Ehrlichia bacteria. Nearly 30% of the screened ticks were PCR-positive; 89% of tick species were PCR-positive, and 88% of animal species carried at least one PCR-positive tick. We sequenced a subset of amplicons that were similar to three Anaplasma species and three Ehrlichia species. The odds of a tick being PCR-positive increased when many ticks were collected from the host or the tick was collected from a cow (domesticated animal). Tick species shared host species more often than expected. We demonstrate that ticks in northwestern Zimbabwe present a One Health problem for nearby wildlife and humans.

Community efforts to monitor and manage Aedes mosquitoes (Diptera: Culicidae) with ovitraps and litter reduction in east Tennessee.

BACKGROUND: East Tennessee (USA) is burdened by mosquito-borne La Crosse virus disease, but minimal resources for mosquito surveillance, management, or related community education exist in the region. To address these needs, we developed a program to train middle and high school educators in basic medical entomology. The educators then used their skills in the classroom to teach students about La Crosse virus disease and conduct mosquito collection experiments. As a case study of a potential application of classroom-collected data, we also partnered with a local non-profit organization to assess the potential for a volunteer litter cleanup to reduce mosquito populations in a Tennessee neighborhood. METHODS: Our first objective was to investigate the ability for educators and their students (schools) to collect high-quality mosquito surveillance data. In 2019 and 2020, we collected Aedes (Diptera: Culicidae) eggs during the same study period as schools and assessed whether data collected by schools reflected the same findings as our own data. Our second objective was to investigate the impact of a volunteer litter cleanup event on Aedes mosquito abundance. In 2021, we collected Aedes eggs before and after a neighborhood trash cleanup while schools conducted their own mosquito egg collections. Using the school collections as non-treatment sites, we used a Before-After-Control-Impact analysis to determine if there was a significant decline in egg abundance after the cleanup. RESULTS: In 2019, mosquito abundance trends were similar between our data and school data but differed significantly during some weeks. After refining our protocols in 2020, school data was highly similar to our data, indicating that schools consistently collected high-quality surveillance data in the program's second year. In 2021, we found a significant decline in Aedes egg abundance after the litter cleanup event in comparison to the schools, but the number of adults reared from those eggs did not differ between sites after the cleanup. CONCLUSION: The results of our work demonstrate the potential for community-driven programs to monitor mosquito abundance trends and for volunteer-based cleanup events to reduce the burden of Aedes mosquitoes. In the absence of infrastructure and resources, academic-community partnerships like the ones evaluated here, provide opportunities to help resource limited areas.

Specifying Pathogen Associations of Amblyomma maculatum (Acari: Ixodidae) in Western Tennessee.

Amblyomma maculatum Koch (Acari: Ixodidae) is established in western Tennessee, a region with increased risk for Rocky Mountain spotted fever and ehrlichiosis. This tick transmits Rickettsia parkeri to humans, likely contributing to cases of rickettsiosis in the region. The objective was to determine pathogen associations within questing and host-collected A. maculatum, and identify ecological factors associated with pathogen infection that may increase the effectiveness of surveillance methods. Of 265 ticks tested, 60 (22.6%) were infected with R. parkeri, and 15 (5.7%) with Candidatus Rickettsia andeanae, a Rickettsia of unknown pathogenicity. Two deer-collected ticks tested positive for Ehrlichia ewingii. No ticks were positive for Anaplasma or Borrelia species. None of the ecological factors tested (collection month, collection source, sex, and habitat type) were associated with R. parkeri infection. This project developed baseline prevalence and incidence data for monitoring pathogen prevalence in A. maculatum populations, and identified an inexpensive method for distinguishing R. parkeri from Ca. R. andeanae.

Comparison of novel and conventional methods of trapping ixodid ticks in the southeastern U.S.A.

Tick-borne disease surveillance and research rely on resource-effective methods for tick collection. This study compared the respective performances of several trapping methods in a mixed grassland-forest habitat in western Tennessee. To test for temporal differences in effectiveness, sites were sampled monthly (April-August 2013) using dry ice, dragging, flagging, sweep netting, carbon dioxide (CO2 ) dragging and CO2 flagging methods. To evaluate the effect of habitat on method effectiveness, four methods (dragging, CO2 dragging, CO2 flagging and dry ice) were compared in four habitat types (bottomland deciduous, upland deciduous, coniferous and grassland) in June 2014. In the temporal comparison, ticks were found to be most abundant in April and May, and there was a significant sampling period and method interaction, such that method effectiveness varied across sampling period. Sweep netting was significantly less effective than the other methods. In the habitat comparison, dry ice trap collections represented the most effective method in upland deciduous and coniferous habitats. Flagging using CO2 was significantly less effective than CO2 dragging and dragging in bottomland deciduous habitats. The success of the various collection methods did not differ significantly within grassland habitats. Overall, dry ice trapping and dragging were the most effective methods for tick collection across time and habitat.

Evaluating Surveillance Methods for Arboviral Vectors of La Crosse Virus and West Nile Virus of Southern Appalachia.

To monitor mosquito-borne diseases, public health departments conduct mosquito and pathogen surveillance. Our objective was to evaluate mosquito monitoring methods for collecting La Crosse virus (LACV) and West Nile virus (WNV) vectors (Aedes and Culex mosquitoes, respectively) in southern Appalachia. Centers for Disease Control and Prevention (CDC) light traps baited with carbon dioxide (CO(2)), CDC light traps baited with CO(2) and BG lure, BG-Sentinel traps baited with CO(2), gravid traps baited with oak (Quercus)-water infusion, and resting traps were compared in eastern Tennessee in 2013. Traps operated at 8 different urban sites throughout Knox County were randomly assigned to and rotated among 6 plots within each site. Results were specific for each vector; the BG-Sentinel trap was the best method for Aedes triseriatus, the CDC trap baited with CO(2) and BG lure was the best method for Ae. albopictus, and the gravid trap was the best method for Ae. japonicus. Culex erraticus collections varied by week and trapping method, indicating no single method was best, but the questing traps collected more mosquitoes. There was no significant trapping difference for Cx. pipiens complex in this region using the methods tested. The results suggest using a combination of trapping methods when sampling for LACV and/or WNV mosquito vectors in southern Appalachia. Effective trapping methods are necessary to enable accurate surveillance, improve control methods, enhance understanding of dispersal, and use for early detection of vectors and pathogens.

Molecular Detection of Rickettsia Species Within Ticks (Acari: Ixodidae) Collected from Arkansas United States.

Rocky Mountain spotted fever (RMSF), caused by the etiological agent Rickettsia rickettsii, is the most severe and frequently reported rickettsial illness in the United States, and is commonly diagnosed throughout the southeast. With the discoveries of Rickettsia parkeri and other spotted fever group rickettsiae (SFGR) in ticks, it remains inconclusive if the cases reported as RMSF are truly caused by R. rickettsii or other SFGR. Arkansas reports one of the highest incidence rates of RMSF in the country; consequently, to identify the rickettsiae in Arkansas, 1,731 ticks, 250 white-tailed deer, and 189 canines were screened by polymerase chain reaction (PCR) for the rickettsial genes gltA, rompB, and ompA. None of the white-tailed deer were positive, while two of the canines (1.1%) and 502 (29.0%) of the ticks were PCR positive. Five different tick species were PCR positive: 244 (37%) Amblyomma americanum L., 130 (38%) Ixodes scapularis Say, 65 (39%) Amblyomma maculatum (Koch), 30 (9%) Rhipicephalus sanguineus Latreille, 7 (4%) Dermacentor variabilis Say, and 26 (44%) unidentified Amblyomma ticks. None of the sequenced products were homologous to R. rickettsii. The most common Rickettsia via rompB amplification was Rickettsia montanensis and nonpathogenic Candidatus Rickettsia amblyommii, whereas with ompA amplification the most common Rickettsia was Ca. R. amblyommii. Many tick specimens collected in northwest Arkansas were PCR positive and these were commonly A. americanum harboring Ca. R. amblyommii, a currently nonpathogenic Rickettsia. Data reported here indicate that pathogenic R. rickettsii was absent from these ticks and suggest by extension that other SFGR are likely the causative agents for Arkansas diagnosed RMSF cases.

Management of Haemaphysalis longicornis (Acari: Ixodidae) on a cow-calf farm in East Tennessee, USA.

Haemaphysalis longicornis Neumann is an exotic and invasive tick species rapidly expanding across the United States. Large infestations of these ticks on cattle can cause anemia, and in eastern regions of the country, they are transmitting Theileria orientalis Ikeda within herds. Methods for managing H. longicornis in the environment are rarely successful and rely on chemicals. To document population effects from producer management decisions, we investigated the change in H. longicornis populations to different tick management practices chosen by producers at 3 different H. longicornis-infested farms. Farm 1 kept a closed herd, chose to cut brush monthly, used on-animal acaricides, and allowed us to drag weekly. Farm 2 kept an open herd, chose to cut brush yearly, did not use on-animal acaricides, and allowed us to drag for ticks weekly/monthly. Farm 3 kept an open herd, chose to cut brush yearly, used on-animal acaracides, and allowed us to drag for ticks monthly. We assessed H. longicornis populations responding to those methods over time using active surveillance methods at each farm. Management decisions on Farms 1 and 3 significantly reduced H. longicornis on farms. Importantly, Farm 1 management decisions reduced the chance of a tick being present by 90%; the number of questing ticks collected fell from 5,000 to 12 H. longicornis in 2 years. Therefore, combined management strategies such as keeping a closed herd, use of on-animal chemical control, mechanical control, and reducing the amount of vegetation on farms were effective ways for producers to manage Haemaphysalis longicornis Neuman (Ixodidae).

Environmental variables serve as predictors of the invasive Asian longhorned tick (Haemaphysalis longicornis Neumann): An approach for targeted tick surveillance.

Since the 2017 discovery of established populations of the Asian longhorned tick, (Haemaphysalis longicornis Neumann) in the United States, populations continue to be detected in new areas. For this exotic and invasive species, capable of transmitting a diverse repertoire of pathogens and blood feeding on a variety of host species, there remains a lack of targeted information on how to best prepare for this tick and understand when and where it occurs. To fill this gap, we conducted two years of weekly tick surveillance at four farms in Tennessee (three H. longicornis-infested and one without) to identify environmental factors associated with each questing life stage, to investigate predictors of abundance, and to determine the likelihood of not collecting ticks at different life stages. A total of 46,770 ticks were collected, of which 12,607 H. longicornis and five other tick species were identified. Overall, abundance of H. longicornis were associated with spring and summer seasons, forested environments, relative humidity and barometric pressure, sunny conditions, and in relation with other tick species. The likelihood of not collecting H. longicornis was associated with day length and barometric pressure. Additional associations for different life stages were also identified and included other tick species, climatic variables, and environmental conditions. Here, we demonstrated that environmental variables can be useful to predict the presence of questing H. longicornis and provide ideas on how to use this information to develop a surveillance plan for different southeastern areas with and without infestations.

Supplemental feeding of deer reduces tick abundance in Mississippi, U.S.A.

Prior research on baiting and feeding of wildlife found changes in habitat and the concentration of wildlife on a local scale (e.g., hundreds of meters). Since changes in habitat and host density affect ticks, feeding wildlife may lead to changes in tick and tick-borne disease ecology. We quantified the effect of feeding deer on ticks and tick-borne diseases at 79 pairs of sites with and without deer feeders during May-August of 2019 and 2020. We captured 0.4 fewer adult (p<0.05) and 1.2 fewer nymphal ticks (p=0.01) at feeder sites. This effect intensified over time with one fewer tick trapped at old feeders (≥5 years) compared to new feeders (<5 years, p<0.05). Greater daily wildlife visitation rates (p<0.001) may have allowed questing ticks to encounter hosts more readily. Most collected ticks were Amblyomma americanum (92.8%), a vector of Ehrlichia and Rickettsia pathogens, though prevalence of these pathogens did not differ (p>0.13) at a local scale. Supplemental deer feeding appears to influence ticks, possibly due to decreased tick habitat and increased wildlife use around feeders. Our findings indicate feeding does not lead to increased prevalence of Ehrlichia or Rickettsia bacteria within A. americanum locally.

Costs of Horn Fly (Diptera: Muscidae) Control for Cow-calf Producers in Tennessee and Texas, 2016.

Tennessee and Texas cow-calf producers were surveyed to assess their 2016 expenses for horn fly control methods. Cattle producers who were members of the Texas and Southwestern Cattle Raisers Association and Tennessee cattle producers who have participated in the Tennessee Agricultural Enhancement Program participated in the survey. Average horn fly management costs in Tennessee and Texas were $9.50/head and $12.40/head, respectively. An ordinary least squares regression and quantile regression were estimated to examine how horn fly costs are influenced by producer and farm demographics, seasonality of horn flies, producer horn fly perceptions, and management practices. When controlling for these variables, Tennessee and Texas cattle producers did not spend significantly different amounts on horn fly control methods. Horn fly costs were associated with producer and farm demographics, producer perceptions of horn flies, and management practices. For example, results indicate that horn fly management costs vary depending on a producer's level of education and income. Having Angus cattle and larger herd sizes were associated with lower costs per head spent on horn fly management. Producers who did not consider horn flies to be a problem until greater quantities of flies were present on the animal spent 15% less per head on managing horn flies. In terms of horn fly control methods, feedthrough insecticides increased horn fly costs the most, followed by using ear tags. This is the first known research to estimate horn fly management costs among cattle producers.

A Unique Academic-Government Collaboration Yields First Report of Detailed Habitat Description for Haemaphysalis longicornis (Ixodida: Ixodidae) in Madison County, KY.

Haemaphysalis longicornis Neumann, Asian longhorned tick, was collected in Madison County, Kentucky, United States as part of an ongoing collaborative-tick surveillance project. This is the first collection of this invasive tick that includes ancillary data on habitat and landscape features derived from the USDA Forest Service, Forest Inventory and Analysis program.

The Common Bed Bug (Hemiptera: Cimicidae) Does Not Commonly Use Canines and Felines as a Host in Low-Income, High-Rise Apartments.

The common bed bug (Cimex lectularius L.) is a known pest and an obligate blood-feeding ectoparasite. Bed bugs can feed on warm-blooded animals including humans, bats, poultry, and rabbits, but no research has investigated the use of companion animals (canines and/or felines) as a blood source. This study investigates how long known host DNA could be detected in a bed bug and the prevalence of bed bugs feeding on companion animals. Laboratory-reared bed bugs were fed host blood to determine how long DNA of human, feline, canine, and rabbit blood could be detected up to 21 d postfeeding. Additionally, 228 bed bugs were collected from 12 apartments with pets (6: canine, 5: feline, and 1: canine and feline), characterized as engorged or unengorged, and then screened with host-specific primers to identify the bloodmeal. Host meals of human, rabbit, feline, and canine blood were detected up to 21 d after feeding laboratory strains. All bed bugs died after feeding on the canine blood, but DNA could be detected up to 21 d post feeding/death. Of the field-collected bed bugs analyzed, human DNA was amplified in 158 (69.3%) bed bugs, canine DNA amplified in 7 bed bugs (3.1%), and feline DNA amplified in 1 bed bug (0.4%). Results of this study suggest that bed bugs predominately feed on humans and rarely feed on companion animals when they cohabitate in low-income, high-rise apartments. Additionally, results from this study warrant future investigations into host use by bed bugs in different housing structures and socioeconomic environments.

Spray and Pour-On Acaricides Killed Tennessee (United States) Field-Collected Haemaphysalis longicornis Nymphs (Acari: Ixodidae) in Laboratory Bioassays.

Haemaphysalis longicornis Neumann (Asian longhorned tick) is an exotic and invasive tick species presenting a health and economic threat to the United States (U.S.) cattle industry due to its ability to transmit pathogens and infest hosts in large numbers. The objective of this study was to evaluate available products at causing H. longicornis mortality in a laboratory bioassay. The efficacy of products was evaluated at label rates using H. longicornis nymphs collected from a cattle farm in eastern Tennessee in two different bioassays (spray or dip) against untreated controls. After exposure, ticks were transferred to clean petri dishes and checked for mortality at 0, 1, 2, 3, 4, 21, 24, and 48 h post exposure. No mortality occurred in the untreated controls, whereas all treated ticks were dead within 24 h of exposure (P < 0.0001). These findings support the hypothesis that currently available spray and pour-on products are effective at causing H. longicornis mortality. We conclude that these acaricides can be used as a component to prevent H. longicornis dispersal and for control in the U.S.

No Evidence of Competition Between the Blacklegged Tick (Acari: Ixodidae) and American Dog Tick on the Rodent Host White-Footed Deermouse (Rodentia: Cricetidae) in Southwestern Tennessee.

Investigations that analyze interspecific associations of vectors on their hosts are important for understanding community structure and implementing ways to comprehend mechanisms of pathogen transmission. We assessed the interspecific association of two tick species (Ixodes scapularis Say (Ixodida: Ixodidae) and Dermacentor variabilis Say (Ixodida: Ixodidae)) on the rodent host Peromyscus leucopus Rafinesque (Rodentia: Cricetidae) at the Hobart Ames Research and Education Center in southwestern Tennessee. Of the rodents captured, 95 (63%) had neither species of tick, 6 (4%) had both tick species, 25 (16%) had I. scapularis only, and 26 (17%) had D. variabilis only. A coefficient of association (C7 = -0.08) was calculated, which suggested there was competition between the two species of ectoparasites, but this value was not significant, indicating that there was a neutral relationship between the tick species on P. leucopus. The co-occurrence of both tick species on their host at the same time suggested that the two tick species can occupy the same host and use the same resources without competing.

Spatial-temporal clusters of host-seeking Aedes albopictus, Aedes japonicus, and Aedes triseriatus collections in a La Crosse virus endemic county (Knox County, Tennessee, USA).

A bite from a La Crosse virus (LACV) infected Aedes mosquito can cause La Crosse encephalitis (LACE), which is a neuro-invasive disease that disproportionately affects children under the age of 16 in Southern Appalachia. The three vectors for LACV are Aedes albopictus (Skuse), Ae. japonicus (Theobald), and Ae. triseriatus (Say). Localized maps of the geographic distribution of vectors are practical tools for mosquito management personnel to target areas with high mosquito abundance. This study hypothesized that LACV vectors have unique species-specific spatial and temporal clusters. To test this, 44 sites were identified in Knox County, Tennessee for their land use/type. At each site, host-seeking mosquitoes were collected approximately every other week from May-October 2018. Spatial clusters of host-seeking mosquito collections for each of the three mosquito species were investigated using Kulldorff's spatial scan statistic, specifying a retrospective space-time Bernoulli model. Most vector clusters were identified in south-central Knox County while the seasonality of clusters varied by mosquito species. Clusters of Ae. albopictus were observed throughout the entire study period while clusters of Ae. japonicus and Ae. triseriatus only occurred May-June. The findings indicate that the relative abundance of LACV vectors were more abundant in south-central Knox County compared to the rest of the county. Of interest, these clusters spatially overlapped with previous LACE diagnosed cases. These findings are useful in guiding decisions on targeted mosquito control in Knox County and may be applied to other counties within Southern Appalachia.

Small-mammal characteristics affect tick communities in southwestern Tennessee (USA).

Life histories can influence the degree of parasite infestations on a host. Pressures exerted on hosts based on age and sex convey varying degrees of parasite prevalence due to differences in host lifestyles, but it is not known how interactions between different host traits affect tick numbers. The objective of this study was to determine if host characteristics (e.g., age, sex, weight, and their interactions) affect the mean number of ticks found on small mammals regardless of host species or habitat. Sherman live traps were placed in forest and grass/forb habitats representative of the southeastern United States. After capture, host characteristics were recorded, and hosts were then searched for ticks. A total of 281 small mammals (148 Peromyscus leucopus, 34 P. maniculatus, 76 Sigmodon hispidus, 16 Microtus pinetorum, and 7 Ochrotomys nuttalli) and 610 ticks (488 Dermacentor variabilis, 114 Ixodes scapularis, 1 Amblyomma americanum, and 7 A. maculatum) were collected in this study. Host's age, sex, and weight affected the number of ticks collected from small mammals and significant interaction effects between host traits occurred (weight by sex, weight by age, and sex by age). For instance, female subadult rodents had significantly more ticks compared to female adults, male subadults had significantly fewer ticks compared to male adults, and the number of ticks on a host increased as host body mass increased. These results support the hypothesis that the number of ticks vary on rodent hosts based on life histories and trait interactions. Therefore, understanding the behavioral mechanisms of a host can aid in the management of parasites in the environment.

Collaborative-Tick Surveillance Works: An Academic and Government Partnership for Tick Surveillance in the Southeastern United States (Acari: Ixodidae).

Tick surveillance provides essential information on distributions and encounter frequencies; it is a component of operational activities in public health practice. Our research objectives were a proof-of-concept for collaborative surveillance, which involved establishing an academic and government partnership to enhance tick surveillance efforts. The University of Tennessee (UT) collaborated with United States Department of Agriculture Forest Service, Southern Research Station Forest Inventory Analysis (FIA) in an Occupational Health and Safety partnership. UT provided FIA crews in the southeastern United States with vials containing 80% ethanol (July 2014-November 2017). Crew members were instructed to put all encountered ticks into the vials and return them to FIA headquarters. UT identified all submitted ticks to species and life stage, and screened Amblyomma americanum (L.) for Ehrlichia bacteria using a nested-PCR assay. From the 198 returned vials, 1,180 ticks were submitted, including A. americanum (90.51%; 202 larvae, 503 nymphs, and 363 adults), Dermacentor variabilis Say (7.12%; 1 nymph, 83 adults), Ixodes scapularis (Say) (1.61%; 19 adults), Amblyomma maculatum Koch (0.59%; 1 nymph, 6 adults), and Amblyomma cajennense (Fabricius) (0.17%; 1 nymph, 1 adult). FIA crews encountered A. americanum with Ehrlichia and collection information was used to generate baseline occurrence data of tick encounters. Results indicate that this collaborative-tick surveillance can be improved and used to generate useful data including pathogen detection, and because crews revisit these sites, changes in tick encounters can be monitored.

Cow-Calf Producers' Willingness to Pay for Bulls Resistant to Horn Flies (Diptera: Muscidae).

Horn flies (Haematobia irritans (L.)) have long posed animal health and welfare concerns. Economic losses to the cattle and dairy industries from their blood-feeding behavior include decreased weight gain, loss in milk productivity, and transmission of bacteria causing mastitis in cattle. Horn fly management strategies are labor intensive and can become ineffective due to the horn fly's ability to develop insecticide resistance. Research indicates that for some cattle herds, genetically similar animals consistently have fewer flies suggesting those animals are horn fly resistant (HFR) and that the trait is heritable; however, it is currently unknown if cattle producers value this trait. Tennessee and Texas cow-calf producers were surveyed to estimate their willingness to pay for HFR bulls and to identify the factors affecting their decision to adopt a HFR bull in their herds. Results indicate that Tennessee and Texas cow-calf producers were willing to pay a premium of 51% and 59% above the base price, respectively, for a HFR bull with the intent to control horn flies within their herd. Producer perceptions of horn fly intensities and the HFR trait, along with their pest management practices, were factors that affected Tennessee and Texas producer willingness to adopt a HFR bull. In Texas, demographics of the producers and their farms also had a role. Knowing producers are willing to pay a premium for the HFR bull indicates that producers value the HFR trait and warrants additional research on the development, implementation, and assessment of the trait.

Beefing Up Biosecurity: Survey of Ticks (Acari: Ixodidae) Currently Threatening the Tennessee Beef Cattle Industry, and a Proposed Monitoring Strategy for Invasive Ticks.

Tick-borne diseases are poised to devastate the North American cattle industry if infected ticks invade the country either by importation of an infested-animal or with natural host migration. Our research objectives were to identify sources for invasive-tick monitoring and use those sources to describe seasonal and regional impacts on infestation prevalence and burden of ticks on beef cattle. Throughout the state of Tennessee, we sampled 25% of the total herd size (or 10 animals) at three university-operated research and education centers (RECs) (sentinel sites), six livestock auctions (check-stations), and nine Extension agents at 21 producer locations (tick scouts) from 2015 to 2016. From 1,817 sampled cattle 740 ticks (Acari: Ixodidae) were collected including 573 Amblyomma americanum (L.) (77.4%), 125 A. maculatum Koch (16.9%), 35 Dermacentor variabilis (Say) (4.7%), and 3 Ixodes scapularis (Say) (0.4%). Western and middle Tennessee were significantly different in infestation prevalence and burden of A. maculatum. For A. maculatum and the species total, infestation prevalence and burden were greater in spring than fall. Auctions (check stations) and RECs (sentinels) had the greatest infestation prevalence of A. maculatum, and the greatest burden of A. maculatum and D. variabilis. High-risk locations clustered in western and middle Tennessee, with low-risk locations in middle and eastern Tennessee. Results from this study provide knowledge necessary to initiate control measures, including seasonal phenology and regional distribution of current tick threats. Use of RECs as sentinel sites and routine tick surveillance at livestock auctions serving as check stations should be used for mitigating invasive tick threats.

Correction: The Microbiome of Ehrlichia-Infected and Uninfected Lone Star Ticks (Amblyomma americanum).

[This corrects the article DOI: 10.1371/journal.pone.0146651.].