Transboundary Tick and Tick-Borne Pathogen Threats to Cattle.

Transboundary incursions of ticks and tick-borne pathogens are ever present concerns for US cattle industries. Global trade in livestock and wildlife, historic and emerging transboundary issues with endemic tick populations and pathogens, and migratory bird flyways are pathways of concern. Transboundary challenges are presented for the Asian long-horned tick and Theileria orientalis Ikeda, for 2 cattle fever tick species [Rhipicephalus (Boophilus) annulatus and R (B) microplus] and Babesia bigemina and B bovis, and for the tropical bont tick and Ehrlichia ruminantium.

Science abhors a surveillance vacuum: Detection of ticks and tick-borne pathogens in southern New Mexico through passive surveillance.

Robust tick surveillance enhances diagnosis and prevention of tick-borne pathogens, yet surveillance efforts in the United States are highly uneven, resulting in large surveillance vacuums, one of which spans the state of New Mexico. As part of a larger effort to fill this vacuum, we conducted both active and passive tick sampling in New Mexico, focusing on the southern portion of the state. We conducted active tick sampling using dragging and CO2 trapping at 45 sites across Hidalgo, Doña Ana, Otero, and Eddy counties between June 2021 to May 2022. Sampling occurred intermittently, with at least one sampling event each month from June to October 2021, pausing in winter and resuming in March through May 2022. We also conducted opportunistic, passive tick sampling in 2021 and 2022 from animals harvested by hunters or captured or collected by researchers and animals housed in animal hospitals, shelters, and farms. All pools of ticks were screened for Rickettsia rickettsii, Rickettsia parkeri, Rickettsia amblyommatis, Ehrlichia ewingii, and Ehrlichia chaffeensis. Active sampling yielded no ticks. Passive sampling yielded 497 ticks comprising Carios kelleyi from pallid bats, Rhipicephalus sanguineus from dogs, mule deer, and Rocky Mountain elk, Otobius megnini from dogs, cats, horses, and Coues deer, Dermacentor parumapertus from dogs and black-tailed jackrabbits, Dermacentor albipictus from domesticated cats, mule deer and Rocky Mountain elk, and Dermacentor spp. from American black bear, Rocky Mountain elk, and mule deer. One pool of D. parumapterus from a black-tailed jackrabbit in Luna County tested positive for R. parkeri, an agent of spotted fever rickettsiosis. Additionally, a spotted fever group Rickettsia was detected in 6 of 7 C. kelleyi pools. Two ticks showed morphological abnormalities; however, these samples did not test positive for any of the target pathogens, and the cause of the abnormalities is unknown. Passive surveillance yielded five identified species of ticks from three domestic and six wild mammal species. Our findings update tick distributions and inform the public, medical, and veterinary communities of the potential tick-borne pathogens present in southern New Mexico.

Species identification of adult ixodid ticks by Raman spectroscopy of their feces.

BACKGROUND: Ticks and tick-borne diseases pose significant challenges to cattle production, thus the species identification of ticks and knowledge on their presence, abundance, and dispersal are necessary for the development of effective control measures. The standard method of inspection for the presence of ticks is the visual and physical examination of restrained animals, but the limitations of human sight and touch can allow larval, nymphal, and unfed adult ticks to remain undetected due to their small size and site of attachment. However, Raman spectroscopy, an analytical tool widely used in agriculture and other sectors, shows promise for the identification of tick species in infested cattle. Raman spectroscopy is a non-invasive and efficient method that employs the interaction between molecules and light for the identification of the molecular constituents of specimens. METHODS: Raman spectroscopy was employed to analyze the structure and composition of tick feces deposited on host skin and hair during blood-feeding. Feces of 12 species from a total of five genera and one subgenus of ixodid ticks were examined. Spectral data were subjected to partial least squares discriminant analysis, a machine-learning model. We also used Raman spectroscopy and the same analytical procedures to compare and evaluate feces of the horn fly Haematobia irritans after it fed on cattle. RESULTS: Five genera and one sub-genus at overall true prediction rates ranging from 92.3 to 100% were identified from the Raman spectroscopy data of the tick feces. At the species level, Dermacentor albipictus, Dermacentor andersoni and Dermacentor variabilis at overall true prediction rates of 100, 99.3 and 100%, respectively, were identified. There were distinct differences between horn fly and tick feces with respect to blood and guanine vibrational frequencies. The overall true prediction rate for the separation of tick and horn fly feces was 98%. CONCLUSIONS: Our findings highlight the utility of Raman spectroscopy for the reliable identification of tick species from their feces, and its potential application for the identification of ticks from infested cattle in the field.

Detection of winter tick, Dermacentor albipictus, infestations using near infrared reflectance spectroscopy of bovine feces.

The objective of this study was to determine whether artificial infestations of D. albipictus could be detected in cattle using near infrared reflectance spectroscopy of bovine feces (fNIRS) and if detection capability was sensitive to size of tick infestation and phase of on-host stage-specific tick development. Fecal samples were collected daily from six non-infested then later tick-infested Bos taurus yearling heifers who each served as their own control. Cattle with D. albipictus infestations arising from as few as 1000 larvae were identified by fecal chemistry changes using fNIRS technology. In two separate trials, three animal pairs were infested with one of three treatment levels (low: ∼ 1000, medium: ∼ 4000, and high: ∼ 8000) of D. albipictus larvae in a repeated measures experimental design. Trial 1 consisted of tick naïve cattle while Trial 2 consisted of prior tick exposed cattle. Date of drop and daily sum of engorged female ticks were tabulated to characterize each infestation. Cluster, common factor, principal component and MANOVA analyses were used to define and assess fecal spectra changes associated with experimental stages of infestation. Cluster analyses found significant differences in fecal samples for heifer pairs in each treatment level group (low, medium, and high) in Trial 1 and then in Trial 2 from two pre-infestation control periods (outside and inside), three stages of tick development (larval feeding, nymphal feeding, adult feeding), and post-tick recovery periods. Five shifts in fecal chemistry of non-infested and tick-infested periods were identified by six clusters of NIRS fecal spectra measured between 576 and 1126 nm. The PCA's resulted in 97.56% and 97.77% for Trials 1 and 2 respectively of the total variation in the 1050 frequencies being explained by the first three principal components (P1, P2, P3). Results from the MANOVA and the Wilk's Lambda test for both trials showed highly significant evidence (p-values < 0.0001) of a difference in the means of the three principal components across the six Stages. There was significant evidence in Trial 1 (p-values = 0.0067) and Trial 2 (p-values < 0.0001) of a difference between the means of the three principal components across the three levels of tick infestation. These significant pair-wise comparisons reflect developmental phases of tick attachment and blood-feeding that define periods of increasing, peak and declining stress identified in five fecal chemistry shifts defined by six fecal spectral clusters.

Tularemia cases increase in the USA from 2011 through 2019.

Tularemia is a rare but potentially serious bacterial zoonosis, which has been reported in the 47 contiguous states of the USA during 2001-2010. This report summarizes the passive surveillance data of tularemia cases reported to the Centers for Disease Control and Prevention from 2011 through 2019. There were 1984 cases reported in the USA during this period. The average national incidence was 0.07 cases per 100,000 person-years (PY), compared to 0.04 cases per 100,000 PY during 2001-2010. The highest statewide reported case 2011-2019 was in Arkansas (374 cases, 20.4% of total), followed by Missouri (13.1%), Oklahoma (11.9%), and Kansas (11.2%). Regarding race, ethnicity, and sex, tularemia cases were reported more frequently among white, non-Hispanic, and male patients. Cases were reported in all age groups; however, individuals 65 years-old and older exhibited the highest incidence. The seasonal distribution of cases generally paralleled the seasonality of tick activity and human outdoor activity, increasing during spring through mid-summer and decreasing through late summer and fall to winter lows. Improved surveillance and education of ticks and tick- and water-borne pathogens should play a key role in efforts to decrease the incidence of tularemia in the USA.

Modeling Effects of Vertebrate Host Exclosures and Host-Targeted Acaricides on Lone Star Tick (Amblyomma americanum, L.) Infestations.

We used a spatially explicit model to simulate the potential effects of exclosures and acaricides targeted at medium-sized mammalian hosts on the local distribution and abundance of lone star ticks (Amblyomma americanum) within forestlands of the southeastern United States. Both exclosures and acaricides were successful in markedly reducing the densities of all off-host tick life stages inside the treatment areas. Densities dropped to almost zero immediately inside the edges of the exclosures, with noticeably depressed densities extending outward 30 to 60 m from the exclosures, and the simulated exclosures maintained their effectiveness as their sizes were decreased from 4.5 to 2.25 to 0.8 ha. Densities exhibited a smooth gradient across the edges of the acaricide-treated areas, with depressed densities extending ≈100 m outward from the edges, but with perceptible densities extending ≈60 m inward from the edges; thus, the simulated acaricide areas lost their effectiveness as size was decreased to slightly less than one-half the diameter of the activity range of the targeted host. Our simulation results indicated that off-host nymph densities responded to reductions of medium-sized host densities. These results suggest that targeting acaricides at medium-sized hosts may be an effective, and currently under-utilized, method for tick suppression.

Amplification and sequencing of entire tick mitochondrial genomes for a phylogenomic analysis.

The mitochondrial genome (mitogenome) has proven to be important for the taxonomy, systematics, and population genetics of ticks. However, current methods to generate mitogenomes can be cost-prohibitive at scale. To address this issue, we developed a cost-effective approach to amplify and sequence the whole mitogenome of individual tick specimens. Using two different primer sites, this approach generated two full-length mitogenome amplicons that were sequenced using the Oxford Nanopore Technologies' Mk1B sequencer. We used this approach to generate 85 individual tick mitogenomes from samples comprised of the three tick families, 11 genera, and 57 species. Twenty-six of these species did not have a complete mitogenome available on GenBank prior to this work. We benchmarked the accuracy of this approach using a subset of samples that had been previously sequenced by low-coverage Illumina genome skimming. We found our assemblies were comparable or exceeded the Illumina method, achieving a median sequence concordance of 99.98%. We further analyzed our mitogenome dataset in a mitophylogenomic analysis in the context of all three tick families. We were able to sequence 72 samples in one run and achieved a cost/sample of ~ $10 USD. This cost-effective strategy is applicable for sample identification, taxonomy, systematics, and population genetics for not only ticks but likely other metazoans; thus, making mitogenome sequencing equitable for the wider scientific community.

Interaction between anti-tick vaccine and a macrocyclic lactone improves acaricidal efficacy against Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae) in experimentally infested cattle.

The southern cattle fever tick (SCFT) Rhipicephalus (Boophilus) microplus, is considered the most important ectoparasite of livestock in the world because of high financial losses associated with direct feeding and transmission of the hemoparasites Babesia bovis, B. bigemina, and Anaplasma marginale. Unfortunately, SCFT in many parts of the world have evolved resistance to all market-available pesticides thus driving development of new control technologies. Vaccination against ticks using the tick gut protein Bm86 has been shown to be effective against acaricide-resistant ticks. This technique has been successfully implemented in Puerto Rico for the control of acaricide-resistant R. microplus on dairy and beef cattle. Observations from Puerto Rico indicate a potentially positive interaction between anti-tick vaccination when used in conjunction with systemic acaricide treatment. In this project, controlled animal studies were completed directly comparing efficacy of anti-tick vaccination with and without systemic acaricide. Results show that the Bm86 anti-tick vaccine in combination with the macrocyclic lactone, Moxidectin, expressed a synergistic interaction, providing greater and longer efficacy than either treatment alone.

Increasing Incidence of Spotted Fever Group Rickettsioses in the United States, 2010-2018.

Spotted fever group Rickettsia species are intracellular bacteria transmitted by tick or mite vectors and that cause human diseases referred to as spotted fever group rickettsioses, or spotted fevers. In the United States, the most recognized and commonly reported spotted fevers are Rocky Mountain spotted fever (RMSF) (Rickettsia rickettsii), Rickettsia parkeri rickettsiosis, Pacific Coast tick fever (Rickettsia species 364D), and rickettsialpox (Rickettsia akari). In this study, we summarize and evaluate surveillance data on spotted fever cases reported to the Centers for Disease Control and Prevention (CDC) through the National Notifiable Diseases Surveillance System from 2010 to 2018. During this period, there were 36,632 reported cases of spotted fevers with 95.83% (N = 35,104) reported as meeting the case definition as probable and 4.17% (N = 1528) reported as meeting the case definition as confirmed. The average national incidence of total cases, both probable and confirmed, was 12.77 cases per million persons per year. The highest statewide incidence was in Arkansas, with 256.84 per million per year, whereas the lowest incidence occurred in California, with 0.32 per million per year (note that spotted fevers were not notifiable in Hawaii and Alaska). Cases of spotted fevers were reported more frequently among males by gender, White by race, and non-Hispanic by ethnicity. The incidence of spotted fevers increased significantly from 2010 to 2018, but it is uncertain how many of the reported cases were RMSF and how many developed from more moderate spotted fevers. Improvement of the ability to differentiate between spotted fever group Rickettsia species is needed.

Raman-based identification of tick species (Ixodidae) by spectroscopic analysis of their feces.

Ticks are blood-feeding parasites that vector a large number of pathogens of medical and veterinary importance. There are strong connections between tick and pathogen species. Timely detection of certain tick species on cattle can cease the spread of numerous devastating diseases such as Bovine babiesiosis and anaplasmosis. Detection of ticks is currently performed by slow and laborious scout-based inspection of cattle. In this study, we investigated the possibility of identification of tick species (Ixodidae) based on spectroscopic signatures of their feces. We collected Raman spectra from individual grains of feces of seven different species of ticks. Our results show that Raman spectroscopy (RS) allows for highly accurate (above 90%) differentiation between tick species. Furthermore, RS can be used to predict the tick developmental stage and differentiate between nymphs, meta-nymphs and adult ticks. We have also demonstrated that diagnostics of tick species present on cattle can be achieved using a hand-held Raman spectrometer. These findings show that RS can be used for non-invasive, non-destructive and confirmatory on-site analysis of tick species present on cattle.

Bovine fecal chemistry changes with progression of Southern Cattle Tick, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) infestation.

Surveillance for cattle fever ticks is an essential activity in the U.S. Cattle Fever Tick Eradication Program which prevents reestablishment of these tick vectors of the pathogens causing bovine babesiosis. Other methods of detecting tick infested cattle could augment current physical inspection of restrained cattle by program inspectors. The objective of this study was to determine whether a single infestation of ∼5000 Rhipicephalus (Boophilus) microplus larvae induced changes in fecal chemistry that were detectable using near-infrared reflectance spectroscopy (NIRS). Fecal samples were collected daily from 6 tick-infested and 6 non-infested Bos taurus yearling heifers. Each infested animal received ticks from one of 6 different strains of laboratory colonies of R. microplus. Date of drop and daily sum of engorged female ticks were tabulated to characterize each infestation. Cluster, common factor, principal component and MANOVA analyses were used to define and assess fecal spectra changes associated with experimental stages of infestation. Cluster analyses found no significant differences in fecal samples from each of the 6 infested heifers. Two shifts in fecal chemistry of infested animals were identified by three clusters of NIRS fecal spectra. The first cluster was comprised of samples from pre-infestation to 9 days after infestation, a period inclusive of larval tick attachment and feeding. The second cluster was comprised of samples from day 10-22 corresponding to the period of nymphal feeding, adult feeding, and early drop of engorged females. A third cluster was comprised of samples from days 23-46 corresponding to the period of engorged female drop and declining tick numbers. A Tukey-Kramer multiple comparison procedure identified significant differences in fecal spectra between five experimental stages of R. microplus infestation for principal component 1 including pre-infestation to nymphal feeding, pre-infestation to adult feeding, larval feeding to adult feeding, nymphal feeding to adult feeding and nymphal feeding to engorged female drop; for principal component 2 including pre-infestation to nymphal feeding, pre-infestation to adult feeding, and pre-infestation to engorged female drop; and for principal component 3 including pre-infestation to drop, and adult feeding to drop. These significant pair-wise comparisons reflect developmental phases of tick attachment and blood-feeding that define periods of increasing, peak and declining stress identified in two fecal chemistry shifts defined by three fecal spectra clusters. Among non-infested animals, two shifts in fecal chemistry were also detected by three fecal-spectra clusters that occurred in synchrony with those of their tick-infested counterparts. There were no significant differences in principal components or MANOVA analyses between infested and non-infested animals and the pattern of significant pair-wise Tukey-Kramer multiple comparisons for non-infested animals were similar to those of infested animals. This unintended confounding effect is attributed to the manner in which all 12 animals were preconditioned as a group, then isolated in randomly assigned blind stalls in a common barn facility for the study, creating the basis for physiological stress resonance among non-infested animals.

Ornithodoros turicata (Dugès) on a Feral Hog in Real County, Texas, USA.

The potential introduction of African swine fever virus (ASFV) into the US has researchers evaluating routes of potential transmission and the establishment of sylvatic cycles. To help document vector-host associations related to ASFV transmission, the finding of O. turicata on a feral hog in Real County, Texas is reported.

Diversity and distribution of the tick-borne relapsing fever spirochete Borrelia turicatae.

Borrelia turicatae is a causative agent of tick-borne relapsing fever (TBRF) in the subtropics and tropics of the United States and Latin America. Historically, B. turicatae was thought to be maintained in enzootic cycles in rural areas. However, there is growing evidence that suggests the pathogen has established endemic foci in densely populated regions of Texas. With the growth of homelessness in the state and human activity in city parks, it was important to implement field collection efforts to identify areas where B. turicatae and its vector circulate. Between 2017 and 2020 we collected Ornithodoros turicata ticks in suburban and urban areas including public and private parks and recreational spaces. Ticks were fed on naïve mice and spirochetes were isolated from the blood. Multilocus sequence typing (MLST) was performed on eight newly obtained isolates and included previously reported sequences. The four chromosomal loci targeted for MLST were 16S ribosomal RNA (rrs), flagellin B (flaB), DNA gyrase B (gyrB), and the intergenic spacer (IGS). Given the complexity of Borrelia genomes, plasmid diversity was also evaluated. These studies indicate that the IGS locus segregates B. turicatae into four genomic types and plasmid diversity is extensive between isolates. Furthermore, B. turicatae and its vector have established endemic foci in parks and recreational areas in densely populated settings of Texas.

Optimizing long-acting acaricide use for integrated tick management of Rhipicephalus australis-infesting cattle in New Caledonia.

The tick Rhipicephalus australis, formerly known as Rhipicephalus microplus, is the most economically important ectoparasite of livestock in New Caledonia, affecting cattle health and production. Decades of control attempts based on the application of chemical acaricides have exerted a strong selective pressure on R. australis populations, some of which have evolved resistance to these treatments. Research to develop integrated tick control programs is now focused on decreasing applications of chemicals. This study reports the implementation of a method of pasture and herd management involving minimal strategic use of long-acting acaricides, here defined as those having substantial efficacy against larvae for several weeks. Diverse parameters concerning the utilization of long-acting acaricides in association with pasture and herd management on 21 New Caledonian farms over a 5-year period were analyzed to optimize their strategic use. Longer larval acaricidal effect was achieved with a commercial pour-on formulation of fluazuron than with a commercial injectable (subcutaneous) formulation containing 3.15% ivermectin. Pasture and herd management allowed an increase in the delay between a long-lasting acaricide application and the subsequent acaricide treatment from 11.0 weeks to 17.7 weeks. However, if ticks were detected and reported by producers on the day of a long-acting acaricide application, the delay to the following treatment was reduced from 18.5 weeks to 11.2 weeks. The impact of a long-acting acaricide treatment on larval populations in pastures was greatest with a stocking rate of 5 animals per hectare grazing during 1 week. These results provide science-based evidence to cattle producers for adaptive integrated tick management in order to delay the development of acaricide resistance.

A Survey of Tick Surveillance and Control Practices in the United States.

Tickborne diseases are an increasing public health threat in the United States. Prevention and diagnosis of tickborne diseases are improved by access to current and accurate information on where medically important ticks and their associated human and veterinary pathogens are present, their local abundance or prevalence, and when ticks are actively seeking hosts. The true extent of tick and tickborne pathogen expansion is poorly defined, in part because of a lack of nationally standardized tick surveillance. We surveyed 140 vector-borne disease professionals working in state, county, and local public health and vector control agencies to assess their 1) tick surveillance program objectives, 2) pathogen testing methods, 3) tick control practices, 4) data communication strategies, and 5) barriers to program development and operation. Fewer than half of respondents reported that their jurisdiction was engaged in routine, active tick surveillance, but nearly two-thirds reported engaging in passive tick surveillance. Detection of tick presence was the most commonly stated current surveillance objective (76.2%). Most of the programs currently supporting tick pathogen testing were in the Northeast (70.8%), Upper and Central Midwest (64.3%), and the West (71.4%) regions. The most common pathogens screened for were Rickettsia spp. (Rickettsiales: Rickettsiaceae) and bacterial and viral agents transmitted by Ixodes (Acari: Ixodidae) ticks. Only 12% of respondents indicated their jurisdiction directly conducts or otherwise financially supports tick control. Responses indicated that their ability to expand the capacity of tick surveillance and control programs was impeded by inconsistent funding, limited infrastructure, guidance on best practices, and institutional capacity to perform these functions.

Integrated control of the cattle tick, Rhipicephalus australis (Acari: Ixodidae), in New Caledonia through the Pasture and Cattle Management method.

Development of the Pasture and Cattle Management (PCM) method is a priority to control the cattle tick, Rhipicephalus australis, in New Caledonia. The PCM method provides the foundation for sustainable integrated tick control because approximately 95% of cattle ticks in infested pastures are off the host in the non-parasitic life stages, and the practice of treating cattle intensely with chemical acaricides is a risk for the emergence of resistance to these active ingredients in commercial acaricidal products available for veterinary use. Here, we report the findings of an assessment survey to document the utility of the PCM method. Analyses of questionnaire data provided by 21 beef cattle producers describing their management of 37 herds informed how to (1) assess the ability of PCM to reduce acaricide use and (2) prioritize best practices and define recommendations to breeders promoting efficient tick control with minimum acaricide use. Boosted regression tree analysis showed a significant (p = 0.002) reduction of ≈33% in the number of acaricide treatments from 7.9 to 5.3 per year by using PCM. Of the 24 factors identified as potentially affecting acaricide use, six factors accounted for ≈86% of the variability in number of acaricide treatments applied annually. The six most influential factors involved farm characteristics as well as pasture and herd management recommendations. These results demonstrated the usefulness of PCM for integrated control of R. australis infestations while reducing acaricide use to improve cattle production in New Caledonia.

Simulated dynamics of southern cattle fever ticks (Rhipicephalus (Boophilus) microplus) in south Texas, USA: investigating potential wildlife-mediated impacts on eradication efforts.

BACKGROUND: Cattle fever ticks (CFT), Rhipicephalus (Boophilus) annulatus and R. (B.) microplus, are vectors of microbes causing bovine babesiosis and pose a threat to the economic viability of the US livestock industry. Efforts by the Cattle Fever Tick Eradication Program (CFTEP) along the US-Mexico border in south Texas are complicated by the involvement of alternate hosts, including white-tailed deer (Odocoileus virginianus) and nilgai (Boselaphus tragocamelus). METHODS: In the present study, we use a spatially explicit, individual-based model to explore the potential effects of host species composition and host habitat use patterns on southern cattle fever ticks (SCFT, R. (B.) microplus) infestation dynamics and efficacy of eradication schemes. RESULTS: In simulations without eradication efforts, mean off-host larval densities were much higher when cattle were present than when only white-tailed deer and nilgai were present. Densities in mesquite and meadows were slightly higher, and densities in mixed brush were much lower, than landscape-level densities in each of these scenarios. In eradication simulations, reductions in mean off-host larval densities at the landscape level were much smaller when acaricide was applied to cattle only, or to cattle and white-tailed deer, than when applied to cattle and nilgai. Relative density reductions in mesquite, mixed brush, and meadows depended on host habitat use preferences. Shifting nilgai habitat use preferences increasingly toward mixed brush and away from mesquite did not change mean off-host larval tick densities noticeably at the landscape level. However, mean densities were increased markedly in mesquite and decreased markedly in mixed brush, while no noticeable change in density was observed in meadows. CONCLUSIONS: Our results suggest that continued integration of field data into spatially explicit, individual-based models will facilitate the development of novel eradication strategies and will allow near-real-time infestation forecasts as an aid in anticipating and preventing wildlife-mediated impacts on SCFT eradication efforts.

Host Bloodmeal Identification in Cave-Dwelling Ornithodoros turicata Dugès (Ixodida: Argasidae), Texas, USA.

Tick-host bloodmeal associations are important factors when characterizing risks of associated pathogen transmission and applying appropriate management strategies. Despite their biological importance, comparatively little is known about soft tick (Argasidae) host associations in the United States compared to hard ticks (Ixodidae). In this study, we evaluated a PCR and direct Sanger sequencing method for identifying the bloodmeal hosts of soft ticks. We collected 381 cave-associated Ornithodoros turicata near San Antonio, Texas, USA, and also utilized eight colony-reared specimens fed artificially on known host blood sources over 1.5 years ago. We correctly identified the vertebrate host bloodmeals of two colony-reared ticks (chicken and pig) up to 1,105 days post-feeding, and identified bloodmeal hosts from 19 out of 168 field-collected soft ticks, including raccoon (78.9%), black vulture (10.5%), Texas black rattlesnake (5.3%), and human (5.3%). Our results confirm the retention of vertebrate blood DNA in soft ticks and advance the knowledge of argasid host associations in cave-dwelling O. turicata.

Raw pacific biosciences and illumina sequencing reads and assembled genome data for the cattle ticks Rhipicephalus microplus and Rhipicephalus annulatus.

Ticks from the genus Rhipicephalus have enormous global economic impact as ectoparasites of cattle. Rhipicephalus microplus and Rhipicephalus annulatus are known to harbor infectious pathogens such as Babesia bovis, Babesia bigemina, and Anaplasma marginale. Having reference quality genomes of these ticks would advance research to identify druggable targets for chemical entities with acaricidal activity and refine anti-tick vaccine approaches. We sequenced and assembled the genomes of R. microplus and R. annulatus, using Pacific Biosciences and HiSeq 4000 technologies on very high molecular weight genomic DNA. We used 22 and 29 SMRT cells on the Pacific Biosciences Sequel for R. microplus and R. annulatus, respectively, and 3 lanes of the Illumina HiSeq 4000 platform for each tick. The PacBio sequence yields for R. microplus and R. annulatus were 21.0 and 27.9 million subreads, respectively, which were assembled with Canu v. 1.7. The final Canu assemblies consisted of 92,167 and 57,796 contigs with an average contig length of 39,249 and 69,055 bp for R. microplus and R. annulatus, respectively. Annotated genome quality was assessed by BUSCO analysis to provide quantitative measures for each assembled genome. Over 82% and 92% of the 1066 member BUSCO gene set was found in the assembled genomes of R. microplus and R. annulatus, respectively. For R. microplus, only 189 of the 1066 BUSCO genes were missing and only 140 were present in a fragmented condition. For R. annulatus, only 75 of the BUSCO genes were missing and only 109 were present in a fragmented condition. The raw sequencing reads and the assembled contigs/scaffolds are archived at the National Center for Biotechnology Information.

Identification of Host Bloodmeal Source in Ornithodoros turicata Dugès (Ixodida: Argasidae) Using DNA-Based and Stable Isotope-Based Techniques.

The ecology and host feeding patterns of many soft ticks (Ixodida: Argasidae) remain poorly understood. To address soft tick-host feeding associations, we fed Ornithodoros turicata Dugès on multiple host species and evaluated quantitative PCR (qPCR) and stable isotope analyses to identify the vertebrate species used for the bloodmeal. The results showed that a qPCR with host-specific probes for the cytochrome b gene successfully identified bloodmeals from chicken (Gallus gallus L.), goat (Capra aegagrus hircus L), and swine (Sus scrofa domesticus) beyond 330 days post-feeding and through multiple molting. Also, qPCR-based bloodmeal analyses could detect multiple host species within individual ticks that fed upon more than one species. The stable isotope bloodmeal analyses were based on variation in the natural abundance of carbon (13C/12C) and nitrogen (15N/14N) isotopes in ticks fed on different hosts. When compared to reference isotope signatures, this method discerned unique δ13C and δ15N signatures in the ticks fed on each host taxa yet could not discern multiple host species from O. turicata that fed on more than one host species. Given the significance of soft tick-borne zoonoses and animal diseases, elucidating host feeding patterns from field-collected ticks using these methods may provide insight for an ecological basis to disease management.