Cattle Fever Research Articles

Desiccant Dusts, With and Without Bioactive Botanicals, Lethal to Rhipicephalus (Boophilus) microplus Canestrini (Ixodida: Ixodidae) in the Laboratory and on Cattle.

The exotic southern cattle fever tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae), since its eradication from the United States in 1943, made a strong incursion into Texas, beginning 2016. The pest is arguably the most economically detrimental ectoparasite of cattle, Bos taurus L., worldwide. Current R. (B.) microplus control mostly relies on conventional synthetic acaricides to which the ixodid has been developing resistance. Our study demonstrates that commercially available desiccant dust products, with and without bioactive botanical additives, are strongly lethal, when applied dry, against larval R. (B.) microplus in the laboratory, and after being released on dust-treated cattle. Deadzone (renamed Celite 610, a diatomaceous earth product), Drione (silica gel + pyrethrins + piperonyl butoxide synergist), and EcoVia (silica gel + thyme oil), each prophylactically prevented larval R. (B.) microplus from attaching to and feeding on stanchioned calves. Desiccant dust-based products are less likely than conventional synthetic acaricides to decline in terms of efficacy as a result of ixodid resistance, and other desiccant dust advantages, including extended residual, flexibility in terms of application methods, environmental, animal, and human safety, and possible compatibility with organic, or 'green', production systems, are discussed. We anticipate that the desiccant dusts we evaluated, and others not included in this study (e.g., kaolin, perlite, and silica gel) will be effective when used with other control tactics in integrated pest management approaches for controlling R. (B.) microplus (and other ixodid species).

Movement ecology of exotic nilgai antelope: A threat to the re‐emergence of cattle fever ticks in the southern USA

AbstractWildlife plays an important role in the emergence of livestock diseases, and their movements can complicate disease management efforts. One of the most significant vector‐borne diseases of livestock worldwide is bovine babesiosis, spread by cattle fever ticks (CFTs; Rhipicephalus [=Boophilus] microplus and Rhipicephalus [=Boophilus] annulatus). Although CFTs were eradicated from the United States by 1943, bovine babesiosis and CFTs are prevalent in México. Recently, management of CFTs in the Texas–México region has been complicated by the presence of free‐ranging, exotic nilgai antelope (Boselaphus tragocamelus). Nilgai are abundant in this region and are competent hosts for CFTs. The goal of this study was to better understand nilgai movements and space use to inform CFT treatment strategies. We analyzed hourly locations from 40 GPS‐collared nilgai in Cameron County, TX, USA, from April 2019 to September 2020. We assigned each nilgai a movement behavior using the net squared displacement metric. We estimated nilgai home range sizes at different temporal scales (monthly, seasonally, and overall) using Brownian bridge movement models. We observed movement patterns consistent with residency (52.5%), seasonal movers (17.5%), dispersal (5%), and unclassified (25%). Resident nilgai had an average maximum axial distance of 7.8 km, while two young females made separate dispersal movements of about 40 km within a year. Overall, nilgai had large and highly variable home ranges: annual median home range estimate for females was 563 ha (range = 105–1545 ha), and for males, it was 937 ha (range = 221–1602 ha). Peak nilgai movements occurred during crepuscular hours, and median hourly movement for females was 57 m/h, and for males, it was 66 m/h. Nilgai home ranges and long‐distance movements have the potential to overlap multiple ranches, as the typical ranch size in South Texas ranges from 250 to 6000 ha. Resident nilgai were more likely to maintain local infestations of CFT. Dispersal events took place during peak tick season, demonstrating the potential for nilgai to aid in the spread of CFT. Understanding these behaviors will help the CFT Eradication Program develop more efficacious treatment strategies to treat infestations in nilgai.

Presence of Anaplasma phagocytophilum Ecotype I in UK Ruminants and Associated Zoonotic Risk.

Anaplasma phagocytophilum is the causative agent of tick-borne fever in sheep, pasture fever in cattle, and granulocytic anaplasmosis in humans. The increasing prevalence and transboundary spread of A. phagocytophilum in livestock, ticks, and wildlife in the UK poses a potential zoonotic risk that has yet to be estimated. Several ecotypes of A. phagocytophilum show variable zoonotic potential. To evaluate the possible risk associated with the transmission of A. phagocytophilum from ruminants to humans, the ecotype was determined by sequencing the groEL gene from 71 positive blood and tissue samples from UK ruminants. Thirty-four groEL sequences were obtained, fourteen of which were identified in multiple samples. Of the 13 nucleotide polymorphisms identified through pairwise comparison, all corresponded to synonymous substitutions. The subsequent phylogenetic estimation of the relationship with other European/world isolates indicated that all the groEL sequences clustered with other ecotype I sequences. The presence of ecotype I closely reflects that observed in ruminants in continental Europe and suggests a lower risk of zoonotic transmission from this reservoir.

Immunogenicity of a freeze-dried combined vaccine against Rift Valley fever and bovine ephemeral fever in cattle.

The target of vaccination is to encourage a strong, covering and long-lasting immune response against antigens. For achieving these objectives; effective adjuvant and new vaccine strategies are demanded to make the vaccine sufficiently immunogenic to instigate a powerful immune response. This study was completed for elaboration and evaluation of freeze-dried combined vaccine against both Rift Valley fever (RVF) and bovine ephemeral fever (BEF) viruses using different stabilizers. Three formulae were prepared from such vaccine including: formula (1): stabilized with a mixture of 5% Lactalbumin Hydrolysate and 2.5% sucrose, formula (2): stabilized with a mixture of 50% the previous stabilizer and 50% of 1% Carbopol and formula (3): stabilized with 1% Carbopol solution. Samples of the three vaccine formulae were reconstituted on the time of experimental animal vaccination using saponin diluent which acts as an adjuvant for both RVFv and BEFv and as an inactivator BEF virus. The ratio between both viruses in all vaccine formulae was 1:1. All vaccine batches were proved to be free of any foreign contaminants and unharmed for experimentally vaccinated animals. Each of the three groups of calves was vaccinated S/C with 2 ml of a reconstituted vaccine formula and their immune response was evaluated using serum neutralization test. The gained results revealed that the prepared combined freeze-dried vaccine with Carbopol elicited a better humoral immune response than the other two vaccine formulae. It could be recommended to use Carbopol as a stabilizer for the preparation of the aimed vaccine.

A "Dual-Addition" Calcium Fluorescence Assay for the High-Throughput Screening of Recombinant G Protein-Coupled Receptors.

G protein-coupled receptors (GPCRs) represent the largest superfamily of receptors and are the targets of numerous human drugs. High-throughput screening (HTS) of random small molecule libraries against GPCRs is used by the pharmaceutical industry for target-specific drug discovery. In this study, an HTS was employed to identify novel small-molecule ligands of invertebrate-specific neuropeptide GPCRs as probes for physiological studies of vectors of deadly human and veterinary pathogens. The invertebrate-specific kinin receptor was chosen as a target because it regulates many important physiological processes in invertebrates, including diuresis, feeding, and digestion. Furthermore, the pharmacology of many invertebrate GPCRs is poorly characterized or not characterized at all; therefore, the differential pharmacology of these groups of receptors with respect to the related GPCRs in other metazoans, especially humans, adds knowledge to the structure-activity relationships of GPCRs as a superfamily. An HTS assay was developed for cells in 384-well plates for the discovery of ligands of the kinin receptor from the cattle fever tick, or southern cattle tick, Rhipicephalus microplus. The tick kinin receptor was stably expressed in CHO-K1 cells. The kinin receptor, when activated by endogenous kinin neuropeptides or other small molecule agonists, triggers Ca2+ release from calcium stores into the cytoplasm. This calcium fluorescence assay combined with a "dual-addition" approach can detect functional agonist and antagonist "hit" molecules in the same assay plate. Each assay was conducted using drug plates carrying an array of 320 random small molecules. A reliable Z' factor of 0.7 was obtained, and three agonist and two antagonist hit molecules were identified when the HTS was at a 2 µM final concentration. The calcium fluorescence assay reported here can be adapted to screen other GPCRs that activate the Ca2+ signaling cascade.

Mechanical devices for off-host tick surveillance

Standard methods for collecting off-host ticks—flags, drags and leggings—expose the investigator to hazards and are typically less efficient for detection of larvae compared to adults. The potential for using mechanical devices such as vacuums and robots was explored. A vacuum device made by reversing the tube on a leaf blower was tested in a pasture known to be infested with cattle fever ticks. For comparison the same pasture was sampled using leggings and by scratch inspection of the infested cattle. Detection of larvae was markedly episodic by both methods reflecting life cycle periodicity. There was a lag of about 3 weeks between detection of larvae in the pasture and adult ticks on the cattle. Over a year of sampling the vacuum collected greater numbers of larval ticks, but the leggings detected clusters of ticks with somewhat greater frequency.

Tick Control in a Connected World: Challenges, Solutions, and Public Policy from a United States Border Perspective.

Ticks are able to transmit the highest number of pathogen species of any blood-feeding arthropod and represent a growing threat to public health and agricultural systems worldwide. While there are numerous and varied causes and effects of changes to tick-borne disease (re)emergence, three primary challenges to tick control were identified in this review from a U.S. borders perspective. (1) Climate change is implicated in current and future alterations to geographic ranges and population densities of tick species, pathogens they can transmit, and their host and reservoir species, as highlighted by Ixodes scapularis and its expansion across southern Canada. (2) Modern technological advances have created an increasingly interconnected world, contributing to an increase in invasive tick species introductions through the increased speed and frequency of trade and travel. The introduction of the invasive Haemaphysalis longicornis in the eastern U.S. exemplifies the challenges with control in a highly interconnected world. (3) Lastly, while not a new challenge, differences in disease surveillance, control, and management strategies in bordering countries remains a critical challenge in managing ticks and tick-borne diseases. International inter-agency collaborations along the U.S.-Mexico border have been critical in control and mitigation of cattle fever ticks (Rhipicephalus spp.) and highlight the need for continued collaboration and research into integrated tick management strategies. These case studies were used to identify challenges and opportunities for tick control and mitigation efforts through a One Health framework.

First serological evidence of Q fever in large ruminants and its associated risk factors in Punjab, Pakistan

Coxiellosis, also known as Q fever, is a zoonotic disease caused by Coxiella burnetii, a gram-negative bacterium that exerts a significant deleterious impact on the productive and reproductive capabilities of livestock, severely effecting the economics of this sector. In this study, 448 sera samples from cattle (n = 224) and buffalo (n = 224) were collected from 112 farms in Pakistan and examined for antibodies against C. burnetii using an indirect ELISA. Ticks were also collected from these animals. Serological analysis revealed a 23.66% and 27.23% seroprevalence of Q fever in cattle and buffalo, respectively. Odds ratio (OR) analysis of the factors associated with C. burnetii seropositivity was performed, and a multivariable logistic model identified five main variables associated with the seropositivity for coxiellosis. These were: (i) the absence of acaricide use (OR 5.61; 95% CI 2.97–10.94); (ii) the presence of ticks (OR 3.23; 95% CI 1.87–5.69); (iii) the abortion history during the preceding year on the farm (OR 14.96; 95% CI 8.09–29.34); (iv) the presence of sheep and goats (OR 2.47; 95% CI 1.20–5.35); and (v) the absence of a separate parturition area (OR 3.17; 95% CI 1.76–5.86). This study provides new insights into the seroprevalence of Q fever in large ruminants across seven studied districts of Punjab, Pakistan, also providing baseline data to inform improved herd management and on-farm practices for the prevention and control of Q fever in large ruminants in the region. Results of this work suggest that further molecular investigation of coxiellosis is warranted to provide a more thorough evaluation of C. burnetii epidemiology in Pakistan.

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.

Periviscerokinin (Cap2b; CAPA) receptor silencing in females of Rhipicephalus microplus reduces survival, weight and reproductive output

BackgroundThe cattle fever tick, Rhipicephalus (Boophilus) microplus, is a vector of pathogens causative of babesiosis and anaplasmosis, both highly lethal bovine diseases that affect cattle worldwide. In Ecdysozoa, neuropeptides and their G-protein-coupled receptors play a critical integrative role in the regulation of all physiological processes. However, the physiological activity of many neuropeptides is still unknown in ticks. Periviscerokinins (CAP2b/PVKs) are neuropeptides associated with myotropic and diuretic activities in insects. These peptides have been identified only in a few tick species, such as Ixodes ricinus, Ixodes scapularis and R. microplus, and their cognate receptor only characterized for the last two.MethodsExpression of the periviscerokinin receptor (Rhimi-CAP2bR) was investigated throughout the developmental stages of R. microplus and silenced by RNA interference (RNAi) in the females. In a first experiment, three double-stranded (ds) RNAs, named ds680-805, ds956-1109 and ds1102-1200, respectively, were tested in vivo. All three caused phenotypic effects, but only the last one was chosen for subsequent experiments. Resulting RNAi phenotypic variables were compared to those of negative controls, both non-injected and dsRNA beta-lactamase-injected ticks, and to positive controls injected with beta-actin dsRNA. Rhimi-CAP2bR silencing was verified by quantitative reverse-transcriptase PCR in whole females and dissected tissues.ResultsRhimi-CAP2bR transcript expression was detected throughout all developmental stages. Rhimi-CAP2bR silencing was associated with increased female mortality, decreased weight of surviving females and of egg masses, a delayed egg incubation period and decreased egg hatching (P < 0.05).ConclusionsCAP2b/PVKs appear to be associated with the regulation of female feeding, reproduction and survival. Since the Rhimi-CAP2bR loss of function was detrimental to females, the discovery of antagonistic molecules of the CAP2b/PVK signaling system should cause similar effects. Our results point to this signaling system as a promising target for tick control.Graphical

Malignant catarrhal fever in cattle in Spain

AbstractWe report a case of malignant catarrhal fever in cattle in a mixed herd (ovine/bovine). The case occurred in the Basque Country (north of Spain) in April 2021. The infection was confirmed by polymerase chain reaction in a calf that had developed signs consistent with the ‘head and eye’ form of the disease. Just before its appearance, a group of five calves born at the farm (including the affected one), aged between 8 and 12 months, were temporarily housed in a pen adjacent to the sheep shed, and separated by a metallic fence that allowed direct contact. In Spain, there are no data on the incidence of malignant catarrhal fever, as there is no active surveillance programme in place. Furthermore, the disease is uncommonly considered in differential diagnoses.

Nanoencapsulation of 4-Propylguaiacol in β-Cyclodextrin, Ethyl Cellulose, and Polyvinylpyrrolidone

We previously identified 4-propylguaiacol to be a highly potent repellent against Rhipicephalus appendiculatus, which transmits East Coast Fever in cattle. So far, the major method that has been employed for tick control is the use of acaricides, which so far has posed a number of challenges. Encapsulation technology may offer a long-term solution to the existing problems by dispensing the repellent at a controlled rate. 4-Propylguaiacol was encapsulated in various nanoparticles, which included β-cyclodextrin, ethyl cellulose, and polyvinylpyrrolidone. The inclusion of 4-propyl guaiacol in the resulting complexes was confirmed by FT-IR, XRD, and SEM analysis. All the sharp peaks belonging to each of the encapsulating polymers were observed. However, some of the characteristic peaks of 4-propylguaiacol disappeared in the complex formed. The rates and duration of release of 4-propylguaiacol from 0.2 g of each inclusion complex were then compared at 38–40°C every 3 hours for 24 hrs. The observed rates of release for 4-propylguaiacol were 0.396 mg/hr., 0.632 mg/hr., and 0.648 mg/hr. Rate from β-cyclodextrin, ethyl cellulose, and PVP inclusion complexes, respectively. The release rate of 4-propylguaiacol in the β-cyclodextrin complex was more controlled than it was in ethyl cellulose and PVP complexes. This controlled release rate exhibited by the β-cyclodextrin complex in small doses for a relatively long time provides a potential tool for dispensing repellents on cattle to protect them from tick bites.

Evaluation of Repellency of Stop the Bites® Botanical Pesticide to White-Tailed Deer at Corn Feeders

Stop the Bites® botanical pesticide was evaluated for repellent properties against white-tailed deer, Odocoileus virginianus (Zimmerman), at feeders with supplemental corn, Zea mays L., in Cameron County, Texas. The pesticide was tested as a ground treatment around feeders and as spray applied directly to deer as they consumed corn at supplemental feeders. With either treatment, there was no significant difference in numbers of white-tailed deer at the feeders compared to the paired nontreated check. The botanical pesticide may be useful for treatment of cattle fever ticks (Rhipicephalus microplus (Canestrini) and Rhipicephalus annulatus (Say)) on deer at supplemental corn feeders during the hunting season when ivermectin-treated corn is withdrawn, or for treatment using a remotely operated sprayer at fence crossings for tick-infested nilgai, Boselaphus tragocamelus (Pallas).

Sero-Prevalence of Rift Valley Fever and Brucellosis in Cattle of Northern Benin, West Africa

Sero-Prevalence of Rift Valley Fever and Brucellosis in Cattle of Northern Benin, West Africa

A Serosurvey for Ruminant Pestivirus Exposure Conducted Using Sera From Stray Mexico Origin Cattle Captured Crossing Into Southern Texas.

The US Department of Agriculture (USDA), Animal Plant Health Inspection Service (APHIS), Cattle Fever Tick Eradication Program (CFTEP) monitor a quarantine zone along the Texas border to prevent the introduction of stray livestock carrying cattle fever ticks entering the United States from Mexico. Stray cattle collected by CFTEP are checked for ticks and several infectious disease-causing pathogens, but not for bovine viral diarrhea virus (BVDV). BVDV is one of the most economically impactful viruses affecting US cattle producers. BVDV is present in all parts of the world, but it has been demonstrated that another distantly related pestivirus, HoBi-like pestivirus (HoBiPev), can also cause BVD. To date, HoBiPev has not been detected in the United States, but is commonly found in Brazil, and sporadically in Europe and Asia. The objective of the current study was to evaluate the seroprevalence of pestiviruses, with a specific focus on HoBiPev, in stray cattle. Virus neutralization (VN) assay was used to determine seroprevalence (or antibody titers) of BVDV-1, BVDV-2, and HoBiPev. Approximately 50% (67 of 134) of the samples were seropositive for pestiviruses; all 67 positive samples were positive (50%) for BVDV-1, 66 samples of the 67 were positive (49.3%) for BVDV-2, and the same 66 samples of the 67 were also positive (49.3%) for HoBiPev. Due to the antigenic cross-reactivity among Pestiviruses, the comparative antibody against each pestivirus was calculated from all VN-positive samples. Titers were clearly higher against BVDV-1, and only one sample had a titer clearly higher against BVDV-2. No sample had an antibody titer higher for HoBiPev, and while this does not prove the absence of HoBiPev, it does provide evidence that the prevalence of HoBiPev is less predominant than BVDV-1. Additionally, data from these samples provide evidence on the susceptibility of animals that may enter into the United States, with ~50% of the animals seronegative for bovine pestiviruses. This cattle population provides a unique opportunity to evaluate and monitor changes in seroprevalence of economically important cattle diseases affecting the cattle industry.

Validation of a single liquid chromatography-tandem mass spectrometry approach for oxytetracycline determination in bull plasma, seminal plasma and urine.

Oxytetracycline is a broad‐spectrum antibiotic, which inhibits protein synthesis and is generally used for the treatment of pneumonia, shipping fever, leptospirosis and wound infections in cattle and swine. The present work proposes a novel liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for oxytetracycline quantification in bull plasma, seminal plasma and urine, requiring limited sample treatment before analysis. Extraction with trichloroacetic acid followed by dilution of the supernatant in mobile phase proved to be effective in all three matrices, allowing to rapidly process large batches of samples. Sharp and symmetrical peak shape was obtained using a BEH C18 reversed‐phase column in a chromatographic run of just 3.5 min. The mass spectrometer operated in positive electrospray ionization mode and monitored specific transitions for oxytetracycline (461.1 → 425.8) and the internal standard demeclocycline (465.0 → 447.6). The method was validated over concentration ranges suitable for field concentrations of oxytetracycline found in each matrix, showing good linearity during each day of testing (R 2 always >0.99), as also confirmed by analysis of variance (ANOVA) and lack‐of‐fit tests. Excellent accuracy and precision were demonstrated by calculated bias always within ±15% and CV% below 10% at all quality control (QC) levels in the three matrices. Matrix effect and recovery were investigated for both analytes, which showed consistent and comparable behaviour in each matrix. To our knowledge, this is the first validated approach for mass spectrometric determination of oxytetracycline in seminal plasma and urine. The method was successfully applied to samples collected during a pharmacokinetic study in bulls, allowing to assess the oxytetracycline concentration–time profile in plasma, seminal plasma and urine.

The Prevalence and Genetic Diversity of Babesia divergens in Ixodes ricinus Nymphs Collected from Farm- and Woodland Sites in Ireland.

The parasite, Babesia divergens causes redwater fever in cattle and a rare, albeit life-threatening disease in humans. In Ireland, B. divergens has always been considered an important pathogen as the high incidence of redwater fever precluded areas of the country from cattle farming. Moreover a relatively large proportion of human cases were reported here. Red deer (Cervus elaphus), which often harbour babesias that are genetically very similar (if not identical) to B. divergens, are quite widespread. In this study 1369 nymphal Ixodes ricinus ticks collected from various habitats were screened for the presence of B. divergens using TaqMan followed by conventional nested PCR. Fragments of the 18S rRNA gene locus (560 bp) were compared against published Irish B. divergens isolates from cattle, humans and red deer. Overall just 1% of I. ricinus nymphs were infected with B. divergens, with similar infection rates in ticks collected from farm- and woodland. Most (90%) 18S rRNA gene fragments derived from woodland ticks were 100% identical to published sequences from cattle and humans. One differed by a single nucleotide polymorphism (SNP) as did two isolates from ticks collected in bogland. Two isolates derived from nymphs collected in farmland differed by 2 and 4 SNPs respectively.

Medicated corn feeders to disinfest cattle fever ticks, Boophilus (Boophilus) microplus (Acari: Ixodidae), from a suburban population of white-tailed deer, Odocoileus virginianus (Cervidae)

Following its eradication from the USA, the cattle fever tick, Rhipicephalus (Boophilus) microplus (Canestrini), a vector of bovine babesiosis, has made episodic incursions into, and sometimes beyond, an established barrier zone separating tick-free from endemic areas. In large part the incursions involve hosting and transport by wild ungulates, particularly deer and antelope. One approach to disinfest ticks from wild hosts is with food baits medicated to stop parasites. The approach has had mixed success due to factors that have been previously identified with supplemental feeding of wildlife especially competition for the bait, social dominance behavior, and the availability of alternative food sources. Given that not all of the target hosts will intake a therapeutic dose of the medication (ivermectin) at all seasons of the year, an open question is whether the approach is efficacious as a stand-alone treatment or even as part of an integrated program. As detailed in the present study an intensive effort was successful in eradicating a local outbreak of fever ticks.

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.

Vaccination of cattle with synthetic peptides corresponding to predicted extracellular domains of Rhipicephalus (Boophilus) microplus aquaporin\xa02 reduced the number of ticks feeding to repletion

BackgroundThere have been ongoing efforts to identify anti-tick vaccine targets to protect cattle from infestation with cattle fever ticks Rhipicephalus (Boophilus) microplus. Two commercial vaccines based on the tick gut protein Bm86 have had variable effectiveness, which has led to poor acceptance, and numerous studies have attempted to identify vaccine antigens that will provide more consistently effective protection. Transcriptomic analysis of R. microplus led to identification of three aquaporin genes annotated to code for transmembrane proteins involved in the transport of water across cell membranes. Previous work showed that vaccination with full-length recombinant aquaporin 1 (RmAQP1) reduced tick burdens on cattle. Targeted silencing of aquaporin 2 (RmAQP2) expression suggested it might also be a good anti-tick vaccination target.MethodsThree synthetic peptides from the predicted extracellular domains of RmAQP2 were used to vaccinate cattle. Peptides were conjugated to keyhole limpet hemocyanin (KLH) as an antigenic carrier molecule. We monitored the antibody response with ELISA and challenged vaccinated cattle with R. microplus larvae.ResultsThere was a 25% reduction overall in the numbers of ticks feeding to repletion on the vaccinated cattle. Immune sera from vaccinated cattle recognized native tick proteins on a western blot and reacted to the three individual synthetic peptides in an ELISA. The vaccinated calf with the highest total IgG titer was not the most effective at controlling ticks; ratios of IgG isotypes 1 and 2 differed greatly among the three vaccinated cattle; the calf with the highest IgG1/IgG2 ratio had the fewest ticks. Ticks on vaccinated cattle had significantly greater replete weights compared to ticks on controls, mirroring results seen with RNA silencing of RmAQP2. However, protein data could not confirm that vaccination had any impact on the ability of the tick to concentrate its blood meal by removing water.ConclusionsA reduced number of ticks feed successfully on cattle vaccinated to produce antibodies against the extracellular domains of RmAQP2. However, our predicted mechanism, that antibody binding blocks the ability of RmAQP2 to move water out of the blood meal, could not be confirmed. Further study will be required to define the mechanism of action and to determine whether these vaccine targets will be useful components of an anti-tick vaccine cocktail.Graphical