Genome resequencing reveals population divergence and local adaptation of blacklegged ticks in the United States.

Tick vectors and tick-borne disease are increasingly impacting human populations globally. An important challenge is to understand tick movement patterns, as this information can be used to improve management and predictive modelling of tick population dynamics. Evolutionary analysis of genetic divergence, gene flow and local adaptation provides insight on movement patterns at large spatiotemporal scales. We develop low coverage, whole genome resequencing data for 92 blacklegged ticks, Ixodes scapularis, representing range-wide variation across the United States. Through analysis of population genomic data, we find that tick populations are structured geographically, with gradual isolation by distance separating three population clusters in the northern United States, southeastern United States and a unique cluster represented by a sample from Tennessee. Populations in the northern United States underwent population contractions during the last glacial period and diverged from southern populations at least 50 thousand years ago. Genome scans of selection provide strong evidence of local adaptation at genes responding to host defences, blood-feeding and environmental variation. In addition, we explore the potential of low coverage genome sequencing of whole-tick samples for documenting the diversity of microbial pathogens and recover important tick-borne pathogens such as Borrelia burgdorferi. The combination of isolation by distance and local adaptation in blacklegged ticks demonstrates that gene flow, including recent expansion, is limited to geographical scales of a few hundred kilometres.

Prevalence of Borrelia burgdorferi and diversity of its outer surface protein C (ospC) alleles in blacklegged ticks (Ixodes scapularis) in Delaware.

Characterizing the diversity of genes associated with virulence and transmission of a pathogen across the pathogen's distribution can inform our understanding of host infection risk. Borrelia burgdorferi is a vector-borne bacterium that causes Lyme disease in humans and is common in the United States. The outer surface protein C (ospC) gene of B. burgdorferi exhibits substantial genetic variation across the pathogen's distribution and plays a critical role in virulence and transmission in vertebrate hosts. In fact, B. burgdorferi infections that disseminate across host tissues in humans are associated with only a subset of ospC alleles. Delaware has a high incidence of Lyme disease, but the diversity of ospC in B. burgdorferi in the state has not been evaluated. We used PCR to amplify ospC in B. burgdorferi-infected blacklegged ticks (Ixodes scapularis) in sites statewide and used short-read sequencing to identify ospC alleles. B. burgdorferi prevalence in blacklegged ticks varied across sites, but not significantly so. We identified 15 previously characterized ospC alleles accounting for nearly all of the expected diversity of alleles across the sites as estimated using the Chao1 index. Nearly 40% of sequenced infections (23/58) had more than one ospC allele present suggesting mixed strain infections and the relative frequencies of alleles in single infections were positively correlated with their relative frequencies in mixed infections. Turnover of ospC alleles was positively related to distance between sites with closer sites having more similar allele compositions than more distant sites. This suggests a degree of B. burgdorferi dispersal limitation or habitat specialization. OspC alleles known to cause disseminated infections in humans were found at the highest frequencies across sites, corresponding to Delaware's high incidence of Lyme disease.

Where have all the grouse ticks gone? Apparent decline in collections of Haemaphysalis chordeilis Packard.

Haemaphysalis chordeilis Packard, also known as the grouse or bird tick, is a three-host tick native to North America. Literature from the early 20th century reported a widespread distribution of this tick across the US and Canada. As its name implies, ground-dwelling birds such as grouse and quail were frequent hosts, and occasionally large infestations were reported in domestic flocks making it a pest of economic importance. However, after the mid-1900's records of this species appear scarce, and a number of more recent studies of ticks on birds (including ostensibly favored host species) did not detect it. To confirm this perception with data, we conducted a literature search for collection records of this species and compared the records across two eras (pre-1965 and post-1965), finding very few records of H. chordeilis in recent years, despite increased attention brought to this genus by the detection of exotic Haemaphysalis longicornis Neumann populations in the eastern US. We also compiled a list of studies after 1965 that examined appropriate hosts for ectoparasites but failed to find H. chordeilis. We interpret the apparent decline of ticks in the context of documented population declines in several major host species over the same time frame and discuss whether ectoparasite populations should be subject to the same conservation consideration as their hosts.

The Gulf Coast Tick, Amblyomma maculatum (Ixodida: Ixodidae), and Spotted Fever Group Rickettsia in the Highly Urbanized Northeastern United States.

We report the multi-year collection of the Gulf Coast tick, Amblyomma maculatum Koch (Acaridae: Ixodida: Ixodidae) in Staten Island, New York City (NYC) as well as their detection in Brooklyn, NYC, and in Atlantic and Cumberland counties in southern New Jersey, USA. The first and most common detections were of adults, however in Freshkills Park on Staten Island larvae were also collected in a following year. The presence of larvae indicates that adults are successfully finding hosts in Staten Island. While it is still unknown how A. americanum reached Staten Island, immatures of this species often parasitize migratory birds, which are now often seen in Freshkills Park. We describe the landscape features of the area in Staten Island where populations were highest and larvae were detected, which could have facilitated the establishment of A. maculatum. Notably, we also report the presence of human pathogens Rickettsia parkeri in 5/10 (50%) of adults tested and R. felis in 1/24 (4.17%) of larvae tested. In addition to established populations in Staten Island we found evidence of A. maculatum in NJ and other NYC boroughs, suggesting current or future establishment is possible. The failure thus far to detect established populations in these areas may be due to inherent difficulties in detecting low density, spatially heterogeneous incipient populations, which could require targeted surveillance efforts for this species. We discuss the consequences to public health of the establishment of A. maculatum and detection of two additional rickettsial pathogens in the densely populated northeastern United States.

Correction: Why Lyme disease is common in the northern US, but rare in the south: The roles of host choice, host-seeking behavior, and tick density.

[This corrects the article DOI: 10.1371/journal.pbio.3001066.].

Why Lyme disease is common in the northern US, but rare in the south: The roles of host choice, host-seeking behavior, and tick density.

Lyme disease is common in the northeastern United States, but rare in the southeast, even though the tick vector is found in both regions. Infection prevalence of Lyme spirochetes in host-seeking ticks, an important component to the risk of Lyme disease, is also high in the northeast and northern midwest, but declines sharply in the south. As ticks must acquire Lyme spirochetes from infected vertebrate hosts, the role of wildlife species composition on Lyme disease risk has been a topic of lively academic discussion. We compared tick-vertebrate host interactions using standardized sampling methods among 8 sites scattered throughout the eastern US. Geographical trends in diversity of tick hosts are gradual and do not match the sharp decline in prevalence at southern sites, but tick-host associations show a clear shift from mammals in the north to reptiles in the south. Tick infection prevalence declines north to south largely because of high tick infestation of efficient spirochete reservoir hosts (rodents and shrews) in the north but not in the south. Minimal infestation of small mammals in the south results from strong selective attachment to lizards such as skinks (which are inefficient reservoirs for Lyme spirochetes) in the southern states. Selective host choice, along with latitudinal differences in tick host-seeking behavior and variations in tick densities, explains the geographic pattern of Lyme disease in the eastern US.

First glimpse into the origin and spread of the Asian longhorned tick, Haemaphysalis longicornis, in the United States.

Established populations of Asian longhorned ticks (ALT), Haemaphysalis longicornis, were first identified in the United States (US) in 2017 by sequencing the mitochondrial cytochrome c oxidase subunit I (cox1) 'barcoding' locus followed by morphological confirmation. Subsequent investigations detected ALT infestations in 12, mostly eastern, US states. To gain information on the origin and spread of US ALT, we (1) sequenced cox1 from ALT populations across 9 US states and (2) obtained cox1 sequences from potential source populations [China, Japan and Republic of Korea (ROK) as well as Australia, New Zealand and the Kingdom of Tonga (KOT)] both by sequencing and by downloading publicly available sequences in NCBI GenBank. Additionally, we conducted epidemiological investigations of properties near its initial detection locale in Hunterdon County, NJ, as well as a broader risk analysis for importation of ectoparasites into the area. In eastern Asian populations (China/Japan/ROK), we detected 35 cox1 haplotypes that neatly clustered into two clades with known bisexual versus parthenogenetic phenotypes. In Australia/New Zealand/KOT, we detected 10 cox1 haplotypes all falling within the parthenogenetic cluster. In the United States, we detected three differentially distributed cox1 haplotypes from the parthenogenetic cluster, supporting phenotypic evidence that US ALT are parthenogenetic. While none of the source populations examined had all three US cox1 haplotypes, a phylogeographic network analysis supports a northeast Asian source for the US populations. Within the United States, epidemiological investigations indicate ALT can be moved long distances by human transport of animals, such as horses and dogs, with smaller scale movements on wildlife. These results have relevant implications for efforts aimed at minimizing the spread of ALT in the United States and preventing additional exotic tick introductions.

Surveillance for Amblyomma maculatum (Acari: Ixodidae) and Rickettsia parkeri (Rickettsiales: Rickettsiaceae) in the State of Delaware, and Their Public Health Implications.

Amblyomma maculatum Koch is the primary vector of Rickettsia parkeri, the etiologic agent of tidewater spotted fever, and can also carry and transmit a variety of other pathogens. This tick historically has been a costly nuisance to livestock owners in the southeastern United States. Over the past 6 yr, A. maculatum has been collected in numbers sufficient to demonstrate their establishment in Kent County, Delaware, and the presence of R. parkeri has been documented. Our goals were to determine the geographic distribution of A. maculatum and R. parkeri in Delaware, and to equate this to relative risk to the public of encountering R. parkeri-infected ticks. We surveyed for ticks in four locations throughout the state from May to August 2019, and found established A. maculatum populations in all three counties. Laboratory analysis of collected specimens by quantitative polymerase chain reaction detected R. parkeri in A. maculatum populations across the state. These results indicate that A. maculatum could present a health risk to inhabitants of the state, and they also historically have posed a risk to the livestock industry, making them an important consideration in the development and implementation of continued tick surveillance efforts and future policies regarding tick management.

Effects of Deltamethrin Treatment on Small Mammal and Ectoparasite Population Dynamics and Plague Prevalence in a North American Mixed-Grass Prairie System.

Sylvatic plague affects many species in North American prairie ecosystems. Deltamethrin is commonly used to manage fleas in potential outbreak areas. Understanding the role of small mammals and their ectoparasites in sylvatic plague maintenance is pertinent to understanding the ecology of plague and its persistence in nature. This study examined the effects of plague management using deltamethrin on communities of small mammals, their flea faunas, and Yersinia pestis prevalence. We trapped small mammals from 2014 to 2016 on the Lower Brule Indian Reservation (LOBR), South Dakota, and analyzed the effects of deltamethrin treatment on small mammal populations, flea loads, and Y. pestis prevalence. We collected higher flea loads from small mammals on sites not treated with deltamethrin (1.10 fleas per animal) than from deltamethrin-treated sites (1.03 fleas per animal). We observed significant negative trends in mean flea load per animal between pre- and post-treatment collections. We detected no significant effects of deltamethrin treatment on animal captures pre- and post-treatment, but observed significant differences in animal captures by experimental unit. We detected no serological evidence for the presence of Y. pestis antibodies in small mammals and 1.2% Y. pestis prevalence across all sampled fleas. Although there is little overlap in the species of fleas infesting small mammals and prairie dogs, the occurrence of flea spillover has been documented. In our study, treatment with deltamethrin reduced flea loads on small mammals by up to 49%. Our data suggest that although the efficacy of deltamethrin on the LOBR-a mixed-grass system-may not be as high as that found in a comparable study in a short-grass system, deltamethrin is still a useful tool in the management of plague.

Surveillance for Ixodes scapularis (Acari Ixodidae) and Borrelia burgdorferi in Eastern South Dakota State Parks and Nature Areas.

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

Flea and Small Mammal Species Composition in Mixed-Grass Prairies: Implications for the Maintenance of Yersinia pestis.

Maintenance of sylvatic plague in prairie dogs (Cynomis spp.) was once thought unlikely due to high mortality rates; yet more recent findings indicate that low-level enzootic plague may be maintained in susceptible prairie dog populations. Another hypothesis for the maintenance of sylvatic plague involves small mammals, other than prairie dogs, as an alternative reservoir in the sylvatic plague system. These hypotheses, however, are not mutually exclusive, as both prairie dogs and small mammals could together be driving sylvatic cycles of plague. The concept of a bridging vector has been used to explain the transmission of pathogens from one host species to another. In the case of sylvatic plague, this would require overlap in fleas between small mammals and prairie dogs, and potentially other species such as carnivores. Our goal was to evaluate the level of flea sharing between black-tailed prairie dogs (Cynomis ludovicianus) and other small mammals in a mixed-grass prairie in South Dakota. We investigated the species richness of small mammals and small-mammal fleas in a mixed-grass prairie system and compared findings with previous studies from a short-grass ecosystem in Colorado. Over the summer field seasons 2014-2016 we live-trapped small mammals, collected fleas, and showed differences between both the flea and small mammal composition of the two systems. We also recorded higher densities of deer mice and lower densities of northern grasshopper mice in mixed versus shortgrass prairies. We confirmed, as is the case in shortgrass prairies, a lack of substantial flea species overlap on small mammal hosts and fleas from prairie dogs and their burrows. Moreover this study demonstrates that although small mammals may not play a large part in interepizootic plague cycling in shortgrass prairie ecosystems, their role in mixed-grass prairies requires further evaluation.

First Evidence of an Established Population of Ixodes scapularis (Acari: Ixodidae) in South Dakota.

Ixodes scapularis Say (Acari: Ixodidae) is the most important vector of human disease in the United States. Recent efforts by public health officials to determine its presence on a county-by-county basis have been undertaken to assist in Lyme disease risk assessment. Recent modeling efforts show that South Dakota can potentially support populations of I. scapularis based on favorable climatic conditions and presence of suitable hosts to support tick populations within the state. We provide the first documentation of an established population of I. scapularis in Clay County, SD, providing only the third record of the presence of this tick species within the state.

Parasitology, virology, and serology of free-ranging coyotes (Canis latrans) from central Georgia, USA.

We examined 31 free-ranging coyotes (Canis latrans) from central Georgia, USA, for select parasites and viral agents. Sixteen coyotes had adult heartworms (Dirofilaria immitis). Serum samples from 27 animals revealed antibodies against canine parvovirus (100%), canine distemper virus (48%), canine adenovirus (37%), and Trypanosoma cruzi (7%); none were detected against Leishmania spp. Twenty-two of 24 (92%) coyotes were positive for Toxoplasma gondii. Real-time PCR of feces revealed 32% of coyotes were shedding canine parvovirus, and sequencing revealed type 2b and 2c. Because coyotes could be a spillover host of domestic dog (Canis lupus familiaris) pathogens, studies of the transmission of pathogens between coyotes and domestic dogs are warranted.

Persistence of two Trichomonas gallinae isolates in chlorinated and distilled water with or without organic material.

Trichomonas gallinae is a protozoan parasite commonly found in columbids, passerines, and raptors. In passerines and columbids, trichomonosis causes significant morbidity and mortality associated with contaminated bird feeders and waters. However, there has been little work on the persistence of T. gallinae in water to determine if artificial waters are a likely source of infection for naive birds. To examine drinking water as a source of T. gallinae transmission, we inoculated 1 x 10(6) trichomonads into containers with 500 ml of either distilled or chlorinated water. In addition, we inoculated the same number of trichomonads in distilled or chlorinated water contaminated with 15 g organic matter. Aliquots of 0.5 ml were collected from each container at 0, 0.5, 1, 5, 10, or 20 min; inoculated into a Trichomonas culture packet; and incubated at 37 C for 6 days. Survival was best in the presence of organic matter, with either distilled or chlorinated water. Uncontaminated chlorinated water did not allow survival at any sampling period.