Kansas beef bulls with chronic anaplasmosis demonstrate satisfactory breeding soundness outcomes at breeding soundness examination.

OBJECTIVE: The objectives of this study were to evaluate the prevalence of chronic Anaplasma marginale infection in beef bulls from eastern Kansas and compare breeding soundness parameters between A marginale-infected and uninfected bulls. We hypothesized that bulls with chronic anaplasmosis would have inferior breeding soundness exam (BSE) outcomes as a result of persistent A marginale infection or the consequence of initial clinical disease compared to uninfected bulls. ANIMALS: 535 client-owned beef bulls from eastern Kansas undergoing routine BSE. METHODS: Complete BSEs were conducted by participating veterinarians according to the second edition of the Society for Theriogenology Manual for Breeding Soundness Examination of Bulls. Blood samples were collected for PCV determination and analysis of A marginale infection status via quantitative PCR and cELISA. Logistic and linear regression methods were used to evaluate factors associated with A marginale infection status and BSE parameters. RESULTS: Prevalence of chronic A marginale infection was 46% (245/535) among bulls. Unsatisfactory BSE outcome was not statistically associated with chronic anaplasmosis in this study population, although more bulls with chronic anaplasmosis had unsatisfactory BSE outcomes (15.0 ± 2.4% vs 12.0 ± 2.2%). CLINICAL RELEVANCE: Chronic anaplasmosis is prevalent among eastern Kansas breeding bulls; however, no negative association between chronic anaplasmosis and breeding soundness at time of BSE was observed.

High levels of alpha-gal with large variation in the salivary glands of lone star ticks fed on human blood.

Tick bites, associated with the secretion of tick saliva containing the xenoglycan galactose-alpha-1, 3-galactose (alpha-gal or aGal), are recognized as the causal factors of alpha-Gal syndrome (AGS; or red meat allergy) in humans. AGS occurs after the increased production of IgE antibodies against aGal, which is found in most mammalian cells, except for the Old World monkey and humans. The aGal sensitization event has been linked to an initial tick bite, followed by consumption of red meat containing the aGal glycan, which triggers the onset of the allergic response resulting in urticaria, anaphylaxis, or even death. In North America, the lone star tick, Amblyomma americanum, has been identified as the main culprit for AGS. However, only a subset of the human population exposed to lone star tick bites develops AGS. This suggests the presence of unidentified variables associated with the sensitization event. To evaluate the quantitative variations of the aGal in ticks, we evaluated the differences in aGal levels in different strains of A. americanum ticks partially fed on different blood sources using an artificial feeding system and animal hosts. We found significantly higher aGal levels in the female ticks fed on human blood than those fed on the blood of other mammals with large variations among different tick populations and individuals. We propose that host-specific genetic components in the A. americanum ticks are involved in the production of high aGal epitope in the tick saliva, which provides a part of the explanation for the variables associated with the AGS sensitization event of the tick bite.

Comparison of the initial and residual speed of Ixodes scapularis kill on dogs treated with a single dose of Bravecto® Chew (25 mg/kg fluralaner) or Simparica TRIO® (1.2 mg/kg sarolaner, 24 µg/kg moxidectin, 5 mg/kg pyrantel).

BACKGROUND: Compliant ectoparasiticide product use is a comprehensive way to control ticks and reduce the risk of tick-borne pathogen transmission to dogs. Because the systemically acting isoxazoline ectoparasiticides require tick attachment for drug delivery, fast speed of kill is essential to minimize tick-borne pathogen transmission risk. METHODS: Dogs of satisfactory tick-carrying capacity were randomly allocated to treatment groups and administered, per label instructions, Bravecto® Chews (minimum 25 mg/kg fluralaner), Simparica TRIO® (minimum 1.2 mg/kg sarolaner, 24 µg/kg moxidectin, 5 mg/kg pyrantel), or no treatment. Dogs were infested with approximately 50 unfed adult (35 female, 15 male) Ixodes scapularis on Day -2, 21 and 28. Live tick counts were performed at 4, 8, 12 and 24 h post-treatment (Day 0) and post-infestation on Day 21 and 28. Tick control efficacy was determined by comparing live tick means for each product-treated group to the untreated control group and each other at all time points using a linear mixed model. The percent of dogs free of live ticks was analyzed using the Fisher's exact test for treatment group comparison. RESULTS: The untreated control group maintained adequate tick infestations throughout the study. Using geometric means, an existing I. scapularis infestation was controlled by 99.7% and 93.0% 12 h post-treatment and by 100% and 99.5% 24 h post-treatment, for Bravecto® and Simparica TRIO®-treated dogs, respectively. Ixodes scapularis infestations were controlled more quickly for Bravecto®- compared to Simparica TRIO®-treated dogs on Day 21 at 8 h (efficacy 74.0% vs. 0.0%, p = 0.003) and 12 h (efficacy 99.2% vs. 39.4%, p < 0.001) post-infestation and Day 28 at 8 h (efficacy 92.2% vs. 0.0%, p < 0.001) and 12 h (efficacy 99.6% vs. 27.7%, p < 0.001) post-infestation. On Day 28 post-treatment, the efficacy of Bravecto® and Simparica TRIO® to control a new I. scapularis infestation was 100% and 96.6%, respectively, by 24 h post-infestation. Of product-treated dogs, 100% of Bravecto®-treated dogs were free of live ticks by 24 h post-treatment or post-infestation. No treatment-related adverse reactions occurred during the study. CONCLUSIONS: Ixodes scapularis infestations are controlled more quickly 21 and 28 days post-treatment for dogs administered a single dose of Bravecto® compared to dogs administered a single dose of Simparica TRIO®.

No evidence for a negative association between bovine leukemia virus status and fertility in Kansas beef herds: a cross-sectional study.

OBJECTIVE: Determine the association between bovine leukemia virus (BLV) status and fertility in beef cows. BLV-status was defined using 3 different testing strategies (ELISA-, quantitative polymerase chain reaction- [qPCR], and high proviral load [PVL]-status). Fertility was defined as the overall probability of pregnancy as well as the probability of becoming pregnant in the first 21 days of the breeding season. ANIMALS: Convenience sample of 2,820 cows from 43 beef herds. PROCEDURES: The association of BLV-status with the probability of becoming pregnant was evaluated with a multivariable logistic regression analysis that used pregnancy status as a binary outcome, herd nested within ranch as a random effect, and BLV-status (ELISA-, qPCR-, and PVL-status as separate models) and potential covariates (eg, age, Body Condition Score [BCS] category, and interactions) as fixed effects. RESULTS: Raw data revealed that 55% (1,552/2,820) of cows were classified as BLV-positive by ELISA, and 95.3% (41/43) of herds had a least 1 ELISA-positive cow. Classification as BLV ELISA-positive was positively associated with the probability of being pregnant; however, when qPCR or PVL were used to classify BLV-status, there was no association with the probability of being pregnant. None of the methods of classifying BLV-status were associated with the probability of becoming pregnant in the first 21 days of the breeding season. CLINICAL RELEVANCE: This study did not find evidence that testing beef cows for BLV-status using ELISA, qPCR, or a cut-off of 0.9 PVL and removing test-positive cows will improve cowherd fertility as described by the probability of becoming pregnant during the breeding season or becoming pregnant during the first 21 days of the breeding season.

Cytauxzoon felis: An Overview.

Cytauxzoon felis is a tick-transmitted, obligate, hemoprotozoal, piroplasmid pathogen of felids and the causative agent of cytauxzoonosis. It has a complex life cycle which includes a tick as its definitive host and a felid as its intermediate host. Since its first description in 1976, C. felis infections of felids have been reported in several southeastern and south-central U.S. states, overlapping with the ranges of its two known biological vectors, Amblyomma americanum (Lone star tick) and Dermacentor variabilis (American dog tick). Infected felids demonstrate disease as either an acute, often-fatal, infection, or a subclinical carrier infection. To develop effective C. felis transmission control strategies, the incidence of acute cytauxzoonosis, patient risk factors, the role of domestic cat carriers, and ecological variabilities need to be investigated further. Of equal importance is communicating these strategies for high-risk cat populations, including recommending year-round use of an acaricide product for all cats that spend any time outdoors. More studies are needed to further identify factors affecting C. felis and other Cytauxzoon spp. infection, transmission, disease progression, and treatment options and outcomes within the U.S. and globally. Here we provide an overview of C. felis highlighting its lifecycle within its definitive host, transmission to its intermediate host, symptoms and signs providing evidence of transmission, definitive diagnosis, current treatment and prevention strategies, and future considerations regarding this condition.

Cross-sectional study to describe bovine leukemia virus herd and within-herd ELISA prevalence and bovine leukemia virus proviral load of convenience-sampled Kansas beef cow-calf herds.

OBJECTIVE: Determine bovine leukemia virus (BLV) seroprevalence of adult female cattle in Eastern Kansas beef herds and the proviral load (PVL) of those cattle found to be ELISA positive. ANIMALS: Convenience sample of 2,845 cows from 44 beef herds. PROCEDURES: BLV serostatus was determined using an ELISA antibody test (gp-51; IDEXX). BLV quantitative PCR (qPCR) status and PVL were determined utilizing a qPCR test (SS1 qPCR test; CentralStar Laboratories). The association of age, herd size, and body condition score (BCS) category on the probability of being BLV positive was evaluated with a multiple variable logistic regression analysis that used BLV status as a binary outcome, herd nested within ranch as a random effect, and BCS, herd size, and age category as fixed effects. RESULTS: Forty-two of 44 herds had at least 1 BLV ELISA-positive cow (95.5% herd seroprevalence). Overall, 1,564 of the 2,845 cows were BLV ELISA positive (55.0% individual animal prevalence). No association between BLV ELISA status and herd size or BCS was identified. When evaluated by age, the model-adjusted probability of being BLV ELISA positive was lowest for heifers (1 year of age, first parity) and increased until 5 to 6 years of age. Of the 1,564 ELISA-positive animals, 838 were qPCR positive (53.6%). The model-adjusted probability of being qPCR positive was not associated with age, herd size, or BCS category. CLINICAL RELEVANCE: This study indicated that BLV-seropositive status both as a herd classification and individual animal classification was very common in this population. Because the percentage of BLV-seropositive cows varied between herds and by age, this study provides evidence that it is essential for investigators to control for herd and age in any analysis of the association of BLV serostatus and health and production outcomes of interest. Some BLV ELSIA-seropositive cows were classified as BLV negative by qPCR, and risk factors may differ between classification status by ELISA and qPCR.

Satisfactory breeding potential is transiently eliminated in beef bulls with clinical anaplasmosis.

BACKGROUND: Natural service breeding is common in U.S. cow-calf operations. Diseases impacting bull reproductive performance have significant economic consequences for producers. Anaplasmosis may be an underappreciated cause of poor reproductive performance in bulls. The primary systemic effects of bovine anaplasmosis including anemia, fever, and weight loss, can all result in unsatisfactory reproductive performance. The objective of this pilot study was to evaluate breeding soundness examination (BSE) outcomes and clinical changes in bulls during and upon resolution of clinical anaplasmosis. Anaplasma marginale-challenged bulls were observed for clinical disease and infection progression and changes in breeding soundness compared to uninfected control bulls for 16 weeks. RESULTS: All Anaplasma marginale-challenged bulls were PCR-positive, seropositive, and showed clinical signs by 3-, 17-, and 24-days post-challenge, respectively. Clinical signs of anaplasmosis included pallor, icterus, fever (≥ 40.2 °C), and weight loss. Acute anemia was observed in all challenged bulls with PCV nadirs ≤ 18% and peak percent parasitized erythrocyte ≥ 50%. Decreased scrotal circumference and poor semen quality (e.g., increased percentage of abnormal spermatozoa, decreased progressively motile sperm), were initially observed within days after onset of clinical anaplasmosis signs and continued weeks beyond disease resolution. Control bulls remained negative for A. marginale. CONCLUSION: This pilot study demonstrates that clinical anaplasmosis reduces breeding soundness in beef bulls. Anaplasmosis should be considered as a differential for bulls with decreased semen quality, especially within endemic areas. A 90 day or greater retest window is recommended for bulls of unsatisfactory breeding potential recently recovered from clinical anaplasmosis.

Comparison of high throughput RNA sequences between Babesia bigemina and Babesia bovis revealed consistent differential gene expression that is required for the Babesia life cycle in the vertebrate and invertebrate hosts.

Bovine babesiosis caused by Babesia bigemina and Babesia bovis is an economically important disease that affects cattle worldwide. Both B. bigemina and B. bovis are transovarially transmitted by Rhipicephalus ticks. However, little is known regarding parasite gene expression during infection of the tick vector or mammalian host, which has limited the development of effective control strategies to alleviate the losses to the cattle industry. To understand Babesia gene regulation during tick and mammalian host infection, we performed high throughput RNA-sequencing using samples collected from calves and Rhipicephalus microplus ticks infected with B. bigemina. We evaluated gene expression between B. bigemina blood-stages and kinetes and compared them with previous B. bovis RNA-seq data. The results revealed similar patterns of gene regulation between these two tick-borne transovarially transmitted Babesia parasites. Like B. bovis, the transcription of several B. bigemina genes in kinetes exceeded a 1,000-fold change while a few of these genes had a >20,000-fold increase. To identify genes that may have important roles in B. bigemina and B. bovis transovarial transmission, we searched for genes upregulated in B. bigemina kinetes in the genomic datasets of B. bovis and non-transovarially transmitted parasites, Theileria spp. and Babesia microti. Using this approach, we identify genes that may be potential markers for transovarial transmission by B. bigemina and B. bovis. The findings presented herein demonstrate common Babesia genes linked to infection of the vector or mammalian host and may contribute to elucidating strategies used by the parasite to complete their life cycle.

Effect of Protracted Free-Choice Chlortetracycline-Medicated Mineral for Anaplasmosis Control on Escherichia coli Chlortetracycline Resistance Profile from Pastured Beef Cattle.

Anaplasmosis is an economically-significant, hemolytic, tick-borne disease of cattle caused by Anaplasma marginale which can cause clinical anemia and death. Current control options are limited, and FDA-approved antimicrobial control options do not have a defined duration of use. A practical and routinely used anaplasmosis control method involves feeding free-choice chlortetracycline (CTC)-medicated mineral to pastured cattle for several months. Constant antimicrobial use poses the risk of expediting the development and dissemination of antimicrobial resistance in off-target commensal bacteria in the bovine gastrointestinal tract. The objective of this study was to determine the CTC-susceptibility of Escherichia coli isolated from anaplasmosis endemic beef cattle herds provided different FDA-approved free-choice CTC-medicated mineral formulations, all intended to provide cattle a dosage of 0.5 to 2.0 mg CTC/lb bodyweight per day. A closed-herd, comprised of Hereford-Angus cows, naturally endemic for anaplasmosis, were grazed in five different pastures with one herd serving as an untreated control group. The other cattle herds were randomly assigned one of four FDA-approved CTC-medicated mineral formulations (700, 5000, 6000, and 8000 g CTC/ton) labeled for "the control of active anaplasmosis" and provided their respective CTC-medicated mineral formulation for five consecutive months. Fecal samples were collected monthly from a subset of cows (n = 6 or 10) per pasture. Fecal samples were cultured for E. coli isolates and the minimal inhibitory concentration of CTC was determined. Baseline CTC-susceptibility of E. coli was variable among all treatment and control groups. The susceptibility of E. coli isolates was significantly different between study herds over the treatment period (p = 0.0037 across time and 0.009 at the final sampling time). The interaction between study herds and treatment period was not significant (p = 0.075).

Failure to Eliminate Persistent Anaplasma marginale Infection from Cattle Using Labeled Doses of Chlortetracycline and Oxytetracycline Antimicrobials.

Bovine anaplasmosis, caused by the intracellular rickettsial pathogen Anaplasma marginale, is the most prevalent tick-transmitted disease of cattle worldwide. In the U.S., tetracycline antimicrobials are commonly used to treat and control anaplasmosis. Oxytetracycline, administered by injection, is indicated for treatment of clinical anaplasmosis in beef and dairy cattle and calves. Chlortetracycline, administered orally, is indicated for control of active anaplasmosis infection in beef and nonlactating dairy cattle. Tetracyclines have been demonstrated to be effective for treating active anaplasmosis, but their ability to eliminate A. marginale at currently approved therapeutic doses or dosing regimens remains unclear. In the absence of approved dosing regimens for A. marginale clearance, a study was conducted to determine the effect of approved oxytetracycline and chlortetracycline indications on A. marginale bacteremia. Fifteen animals with persistent anaplasmosis were enrolled and divided into three treatment groups. Group 1 (n = 6) received oral chlortetracycline (1.1 mg/kg bodyweight) administered via hand-fed medicated feed for 60 consecutive days. Group 2 (n = 6) received injectable oxytetracycline administered subcutaneously at 19.8 mg/kg bodyweight three times in 3-week intervals. Group 3 (n = 3) served as an untreated control. After 60 days, bacteremia failed to permanently decrease in response to treatment. This result indicates that clearance of A. marginale is unlikely to be reliably achieved using currently approved tetracycline-based regimens to manage anaplasmosis.

Early onset of pre-lethal effects of lotilaner (Credelio®) on Amblyomma americanum ticks on experimentally infested dogs.

BACKGROUND: The speed with which acaricides paralyze and kill ticks is relevant to impeding pathogen transmission. The objective of this study was to assess early-onset lotilaner effects on the motility and weights of Amblyomma americanum ticks collected from treated dogs. METHODS: Twelve healthy dogs were randomized between two groups to receive either lotilaner (Credelio®) on Day 0 or to be sham treated. On Day 7, 25 male and 25 female A. americanum were placed under bandages, two on each flank of each dog. After 30 or 45 min, all unattached ticks were removed and T = 0 was set. At T = 2, 4, 8 and 24 h post attachment, 5 attached ticks removed from each bandage on each dog were weighed, assessed by blinded observers for righting ability and movement recorded. RESULTS: After the infestation period significantly fewer treated than control dogs had 20 ticks attached (50.0% versus 91.7%, P = 0.0015). At 24 h post attachment, mean weights of ticks from treated dogs (males 1.69 mg; females 2.72) were significantly less than ticks from controls (males 2.66 mg; females 4.67) (Pmale = 0.0002; Pfemale < 0.0001). Mean tick weights from the treated group were significantly lower at 24 h than at earlier time points (Pmale < 0.0307; Pfemale = 0.0021). At 4 and 8 h, significantly fewer ticks from treated (14.3%, 0.0%, respectively) than from control dogs could right (73.3%, 70.0%) (P4h < 0.0001; P8h = 0.0024) (at 24 h, all ticks from treated dogs were dead), and distance moved was significantly less at all time points (P2h = 0.0413; P4h, P8h < 0.0001). Mean and maximum velocity of ticks from treated dogs were significantly lower, relative to controls, at 4 and 8 h (P ≤ 0.0001). Within the treated group, collected ticks had significantly lower mean and maximum velocities at 4 and 8 h compared to 2 h (Pmean < 0.0042; Pmax < 0.0194). CONCLUSION: The observed changes indicate that lotilaner may disrupt tick attachment. In ticks that attached, a progressive impairment of neuromuscular processes began within 2 h. Those irreversible changes could substantially reduce the risk of pathogen transmission from tick to host.

AC-DC electropenetrography unmasks fine temporal details of feeding behaviors for two tick species on unsedated hosts.

Ticks are significant nuisance pests and vectors of pathogens for humans, companion animals, and livestock. Limited information on tick feeding behaviors hampers development and rigorous evaluation of tick and tick-borne pathogen control measures. To address this obstacle, the present study examined the utility of AC-DC electropenetrography (EPG) to monitor feeding behaviors of adult Dermacentor variabilis and Amblyomma americanum in real-time. EPG recording was performed during early stages of slow-phase tick feeding using an awake calf host. Both tick species exhibited discernable and stereotypical waveforms of low-, medium-, and high-frequencies. Similar waveform families and types were observed for both tick species; however, species-specific waveform structural differences were also observed. Tick waveforms were hierarchically categorized into three families containing seven types. Some waveform types were conserved by both species (e.g., Types 1b, 1c, 2b, 2c) while others were variably performed among species and individually recorded ticks (e.g., Types 1a, 2a, 2d). This study provides a proof-of-principle demonstration of the feasibility for using EPG to monitor, evaluate, and compare tick feeding behaviors, providing a foundation for future studies aimed at correlating specific feeding behaviors with waveforms, and ultimately the influence of control measures and pathogens on tick feeding behaviors.

Comparative analysis of gene expression between Babesia bovis blood stages and kinetes allowed by improved genome annotation.

Throughout their life cycle, Babesia parasites alternate between a mammalian host, where they cause babesiosis, and the tick vector. Transition between hosts results in distinct environmental signals that influence patterns of gene expression, consistent with the morphological and functional changes operating in the parasites during their life stages. In addition, comparing differential patterns of gene expression among mammalian and tick parasite stages can provide clues for developing improved methods of control. Hereby, we upgraded the genome assembly of Babesia bovis, a bovine hemoparasite, closing a 139 kbp gap, and used RNA-Seq datasets derived from mammalian blood and tick kinete stages to update the genome annotation. Of the originally annotated genes, 1,254 required structural changes, and 326 new genes were identified, leading to a different predicted proteome compared to the original annotation. Next, the RNA-Seq data was used to identify B. bovis genes that were differentially expressed in the vertebrate and arthropod hosts. In blood stages, 28% of the genes were upregulated up to 300 fold, whereas 26% of the genes in kinetes, a tick stage, were upregulated up to >19,000 fold. We thus discovered differentially expressed genes that may play key biological roles and serve as suitable targets for the development of vaccines to control bovine babesiosis.

AC-DC electropenetrography: fundamentals, controversies, and perspectives for arthropod pest management.

Studying the intimate association of arthropods with their physical substrate is both important and challenging. It is important because substrate is a key determinant for organism fitness; challenging because the intricacies of this association are dynamic, and difficult to record and resolve. The advent of electropenetrography (EPG) and subsequent developments allowed researchers to overcome this challenge. Nonetheless, EPG research has been historically restricted to piercing-sucking hemipteran plant pests. Recently, its potential use has been greatly broadened for additional pests with instrument advances. Thus, blood-feeding arthropods and chewing feeders, as well as non-feeding behaviors like oviposition by both pests and parasitoids, are novel new targets for EPG research, with critical consequences for integrated pest management. EPG can explain mechanisms of crop damage, plant or animal pathogen transmission, and the effects of insecticides, antifeedants, repellents, or transgenic plants and animals, on specific behaviors of damage or transmission. This review broadly covers the principles and development of EPG technology, emphasizing controversies and challenges remaining with suggested research to overcome them. In addition, it summarizes 60+ years of basic and applied EPG research, and previews future directions for pest management. The goal is to stimulate new applications for this unique enabling technology. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.

Acute Cytauxzoon felis Cases in Domestic Cats from Eastern Kansas, a Retrospective Case-Control Study (2006-2019).

Cytauxzoon felis, a tick-borne hemoprotozoal pathogen of felids, causes an acute, often-fatal disease in domestic cats. While public awareness of the disease has increased, few studies have evaluated the incidence of acute cytauxzoonosis cases and their associated risk factors. The objective of this study was to retrospectively review records of cats diagnosed with acute cytauxzoonosis in eastern Kansas from 2006-2019 using clinic records and determine: (i) feline cytauxzoonosis risk factors; and (ii) if cytauxzoonosis case incidence is increasing. Although inter-annual variation of acute cytauxzoonosis diagnosis was observed in the eastern Kansas domestic cat population, the overall incidence trend remained largely unchanged over the 14-year case review period. In comparison to ill (C. felis-unrelated) control cases, more acute cytauxzoonosis cases were diagnosed in spring and summer, suggesting a seasonal fluctuation of infection, with samples most commonly submitted from ≥1 year old, owned, male cats. Although cytauxzoonosis case submissions remained consistent over the broad study period, increasing tick vector and domestic cat reservoir populations may contribute to additional cytauxzoonosis case expansion in endemic areas. Investigating the incidence of acute cytauxzoonosis, patient risk factors, and ecological variables that influence disease transmission are important steps towards developing and communicating the need for effective cytauxzoonosis control strategies for high-risk cat populations.

Transcriptome dataset of Babesia bovis life stages within vertebrate and invertebrate hosts.

Babesia bovis is a hemoprotozoan parasite of cattle that has a complex life cycle within vertebrate and invertebrate hosts. In the mammalian host, B. bovis undergoes asexual reproduction while in the tick midgut, gametes are induced, fuse, and form zygotes. The zygote infects tick gut epithelial cells and transform into kinetes that are released into the hemolymph and invade other tick tissues such as the ovaries, resulting in transovarial transmission to tick offspring. To compare gene regulation between different B. bovis life stages, we collected parasites infecting bovine erythrocytes and tick hemolymph. Total RNA samples were isolated, and multiplexed libraries sequenced using paired-end 100 cycle reads of a HiSeq 2500. The data was normalized using the TMM method and analysed for significant differential expression using the generalized linear model likelihood ratio test (GLM LRT) in edgeR. To validate our datasets, ten genes were selected using NormFinder. Genes that had no significant fold change between the blood and tick stages in the RNA-Seq datasets were tested by quantitative PCR to determine their suitability as "housekeeping" genes. The normalized RNA-Seq data revealed genes upregulated during infection of the mammalian host or tick vector and six upregulated genes were validated by quantitative PCR. These datasets can help identify useful targets for controlling bovine babesiosis.

Prevalence of Cytauxzoon felis Infection-Carriers in Eastern Kansas Domestic Cats.

Cytauxzoon felis is a hemoprotozoal tick-transmitted pathogen of felids. Felids that survive acute disease often remain infected and serve as reservoirs for subsequent tick transmission to other susceptible felines. States adjacent to Kansas have identified C. felis-domestic cat carriers while statewide awareness and concern of cytauxzoonosis have increased. The objective of this study was to determine the prevalence of C. felis-carriers in the eastern Kansas domestic cat population using a sensitive quantitative PCR assay targeting the C. felis Cox3 mitochondrial gene. An overall C. felis infection prevalence of 25.8% was determined for asymptomatic domestic cats in eastern Kansas. Significantly more C. felis-carrier cats were identified in spring and fall, suggesting a seasonal fluctuation of survivors. Additionally, a greater percentage of feral and owned cats were positive for C. felis compared to rescue/rescinded cats. This study demonstrates that C. felis-domestic cat carriers are common among cats that spend at least a portion of time outdoors in eastern Kansas, and that more cats likely survive cytauxzoonosis than expected. Understanding the role of domestic cat carriers of C. felis is essential in developing cytauxzoonosis mitigation strategies, including recommending year-round use of acaricide products for all cats that spend any time outdoors.

Bovine anaplasmosis herd prevalence and management practices as risk-factors associated with herd disease status.

Bovine anaplasmosis is a hemolytic disease of cattle caused by Anaplasma marginale which can cause anemia, adult mortality, abortion, and performance reduction. The objectives of this study were to estimate herd-level infection prevalence of bovine anaplasmosis in Kansas cow-calf herds and assess management practices associated with herd infection status. Licensed Kansas veterinarians were randomly selected and provided clientele to generate randomly selected participant herds. Blood samples were collected from 10 mature cows during processing of 925 herds between October 1, 2016 and March 1, 2017. A management survey was completed by 780 herd-owners. Sample status was determined by competitive enzyme-linked immunosorbent assay (cELISA); operations indicating vaccination for anaplasmosis were tested with A.marginale-specific polymerase chain reaction (PCR). Survey data underwent logistic regression analysis for calculation of odds ratios and confidence intervals. The herd-level prevalence was 52.5 % of cow-calf herds. Prevalence ranged from 19.1 % of herds in Western Kansas to 87.3 % of herds in Eastern Kansas. Vaccinated herds were more likely (OR = 2.38; CI = 1.16-4.85; p =  0.02) to be positive compared to non-vaccinated herds, and herds that utilized insecticide ear-tags were more likely to be positive (OR = 1.9; CI = 1.42-2.55; p < 0.01) compared to herds which do not. Operations that prescribe-burned 21-50 % and >50 % of their pastures were more likely to be test positive, OR = 5.74 (CI = 3 .14-10.51; p < 0.01) and OR = 4.78 (CI = 2.33-10.17; p < 0.01), respectively, than operations that prescribe-burned <20 % of their pastures. In summary, anaplasmosis is present across Kansas beef herds at varied prevalence levels and selected management practices were found to be associated with herd infection status.

Determination of plasma-chlortetracycline (CTC) concentrations in grazing beef cattle fed one of four FDA approved free-choice CTC-medicated minerals.

Control of active bovine anaplasmosis in the United States is predicated on the use of chlortetracycline (CTC)-medicated feed throughout the vector season. However, data describing population pharmacokinetics of chlortetracycline in cows, on pasture, having free-choice access to CTC-medicated mineral for consecutive months is lacking. This study documented plasma-CTC concentrations in grazing cows during peak vector season in an anaplasmosis endemic herd. Each pasture was administered one of the four Food and Drug Administration approved CTC-medicated mineral formulations and were assigned as follows: 0.77 g/kg, Aureo Anaplaz C700 Pressed (Sweetlix Livestock Supplements, Mankato, MN); 5.5 g/kg, Purina Anaplasmosis Block (Purina Animal Nutrition, Gray Summit, MO); 6.6 g/kg, Stockmaster Aureo FC C6000 Mineral (Hubbard Feeds, Mankato, MN); 8.8 g/kg, MoorMan's Special Range Minerals AU 168XFE (ADM Animal Nutrition, Quincy, IL). Blood samples were collected monthly for determining plasma drug concentration by Ultra performance liquid chromatography (UPLC) and mass spectrometry. Continued plasma-CTC monitoring allowed for characterization of trends between treatment groups (pastures), age groups (<3 yr or >4 yr), and sampling times (June to October). Results indicate formulation (pasture) and time were significant factors affecting concentrations of CTC in plasma. Cows exposed to 5.5 g/kg block formulation recorded higher CTC plasma concentrations compared with other pasture groups (P = 0.037). Plasma-CTC concentrations increased over time (month of measurement; P = 0.0005). Specifically, concentrations measured after 5 months of continuous CTC treatment were higher than those measured in earlier months.