Assessment of cattle tick infestation: Molecular insights into Rhipicephalus annulatus and the efficacy of garlic oil and nanoemulsion as acaricidal agents.

Ticks, particularly Rhipicephalus annulatus, pose significant threats to livestock, causing economic losses and transmitting various infectious diseases. This study aimed to evaluate the potential acaricidal properties of garlic oil and its nanoemulsion against ticks infesting cattle, Rhipicephalus annulatus through the evaluation of mortality rate and morphological changes of the treated ticks. The study also included prevalence, risk factors, and molecular confirmation of tick species. Genetic characterization confirmed the identity of R. annulatus. Our results revealed a high prevalence of R. annulatus (46.9%) with a higher risk in male cattle (50%) than females (44.9%) and a nonsignificant high infection (49.1%) in animals ≤ 1 year old. The acaricidal efficiency of garlic oil and its nanoemulsion was concentration and time-dependent. The high concentration of garlic oil (20 mg/L) induced complete mortality within 48 hours. The nanoemulsion formulation enhanced efficacy, particularly at 5 mg/L, which exhibited rapid and substantial acaricidal activity. Scanning electron microscopy revealed morphological alterations induced by garlic oil and its nanoemulsion, including changes to the anterior capitulum, dorsal, and ventral cuticles. The study contributes to the exploration of effective, safe, and eco-friendly alternatives for tick control. Further research is warranted to validate their efficacy under diverse conditions and assess practical strategies.

Dissipation behavior, residue distribution, and exposure risk assessment of tebufenpyrad and milbemectin acaricides in strawberries under open field conditions.

Strawberries are a favorite fruit for most people, but the residues of pesticides on strawberries might be risky to human health. Tebufenpyrad and milbemectin are broad-spectrum acaricides with insecticide properties authorized for use on strawberries in Egypt. As a result, it is crucial to investigate their residues in the final product to ensure customers' safety. Consequently, field trials were conducted following the Good Agricultural Practices (GAPs) to study the dissipation rate and terminal residues of tebufenpyrad and milbemectin on strawberries. Tebufenpyrad and milbemectin residues in strawberries declined due to first-order decay process, showing significant degradation (88.5% and 94.7%, respectively) after 14 days. Risk assessment study was carried out by comparing the national estimated daily intake (NEDI) to the acceptable daily intake (ADI). The results demonstrated that the dietary risk posed by the residues of tebufenpyrad and milbemectin in strawberry fruits was acceptable for consumers. It is envisaged that the current study's findings would support the safe application of tebufenpyrad and milbemectin to strawberries and perhaps other crops in Egypt and other countries with similar climatic conditions.

Assessment of the in vitro acaricidal activity of Bravecto® (fluralaner) and a proposed orange oil-based formulation vehicle for the treatment of Sarcoptes scabiei.

BACKGROUND: Sarcoptic mange is a serious animal welfare concern in bare-nosed wombats (Vombatus ursinus). Fluralaner (Bravecto®) is a novel acaricide that has recently been utilised for treating mange in wombats. The topical 'spot-on' formulation of fluralaner can limit treatment delivery options in situ, but dilution to a volume for 'pour-on' delivery is one practicable solution. This study investigated the in vitro acaricidal activity of Bravecto, a proposed essential oil-based diluent (Orange Power®), and two of its active constituents, limonene and citral, against Sarcoptes scabiei. METHODS: Sarcoptes scabiei were sourced from experimentally infested pigs. In vitro assays were performed to determine the lethal concentration (LC50) and survival time of the mites when exposed to varying concentrations of the test solutions. RESULTS: All compounds were highly effective at killing mites in vitro. The LC50 values of Bravecto, Orange Power, limonene and citral at 1 h were 14.61 mg/ml, 4.50%, 26.53% and 0.76%, respectively. The median survival times of mites exposed to undiluted Bravecto, Orange Power and their combination were 15, 5 and 10 min, respectively. A pilot survival assay of mites collected from a mange-affected wombat showed survival times of < 10 min when exposed to Bravecto and Orange Power and 20 min when exposed to moxidectin. CONCLUSIONS: These results confirm the acaricidal properties of Bravecto, demonstrate acaricidal properties of Orange Power and support the potential suitability of Orange Power and its active constituents as a diluent for Bravecto. As well as killing mites via direct exposure, Orange Power could potentially enhance the topical delivery of Bravecto to wombats by increasing drug penetration in hyperkeratotic crusts. Further research evaluating the physiochemical properties and modes of action of Orange Power and its constituents as a formulation vehicle would be of value.

Laboratory assessment of the acaricidal, repellent and anti-cholinesterase effects of Melaleuca alternifolia and Chamaemelum nobile essential oils against Hyalomma scupense ticks.

In cattle, Hyalomma scupense serves as an important vector of several pathogens resulting in diseases, subsequently affecting the agricultural field as well as the economy. Resistance to chemical acaricides has become widespread affirming the need for new drugs to tick control. The goal of this study was to investigate the acaricidal, repellent activities as well as the putative mode of action of two essential oils (EOs) from Melaleuca alternifolia (Tea tree) and Chamaemelum nobile (Roman chamomile) on Hyalomma scupense. The chemical composition of EOs was also evaluated. Different concentrations of EOs were tested in vitro for their acaricidal property on adults and larvae of H. scupense using adult immersion test (AIT) and larval packet test (LPT). Additionally, using Ellman's spectrophotometric method, the anticholinesterase (AChE) inhibition activity of M. alternifolia and C. nobile EOs was assessed in order to understand their putative mode of action. The main compounds of C. nobile were α-Bisabolene (22.20%) and (E)-ß-Famesene (20.41%). The major components in the analyzed M. alternifolia were Terpinen-4-ol (36.32%) and γ-Terpinene (13.69%). Adulticidal and larvicidal assays demonstrated a promising efficacy of the essential oils against tick H. scupense. The lethal concentration (LC50) values obtained for M. alternifolia and C. nobile oils were 0.84 and 0.96 mg/mL in the AIT and 0.37 and 0.48 mg/mL in the LPT, respectively. Regarding repellent activity, M. alternifolia achieved 100% repellency at the concentration of 1 mg/mL while C. nobile showed 95.98% repellency activity at concentration of 4 mg/mL. Also, M. alternifolia and C. nobile EOs displayed potent AChE inhibition with IC50 value of 91.27 and 100.12 µg/mL, respectively. In the present study, M. alternifolia and, to a lesser degree, C. nobile EOs were found to be effective in vitro acaricides, repellents and acetylcholinesterase inhibitor against H. scupense ticks. These plants may represent an economical and sustainable alternative to toxic synthetic acaricides in the management of ectoparasites of veterinary importance.

Community-based integrated tick management programs: cost and feasibility scenarios.

Numerous studies have assessed the efficacy of environmentally based control methods to suppress populations of the blacklegged tick (Ixodes scapularis Say), but few of these estimated the cost of control. We estimated costs for a range of tick control methods (including habitat management, deer exclusion or population reduction, broadcast of acaricides, and use of host-targeted acaricides) implemented singly or in combination and applied to a model community comprising 320 residential properties and parklands. Using the high end for cost ranges, tick control based on a single method was estimated to have mean annual costs per household in the model community ranging from $132 for treating only forest ecotone with a broadcast synthetic acaricide to kill host-seeking ticks (or $404 for treating all residential forested habitat) to >$2,000 for deployment of bait boxes (SELECT TCS) across all residential tick habitat to treat rodents topically with acaricide to kill infesting ticks. Combining different sets of multiple methods in an integrated tick management program placed the annual cost between $508 and 3,192 annually per household in the model community, underscoring the disconnect between what people in Lyme disease endemic areas say they are willing to pay for tick control (not more than $100-150 annually) and the actual costs for tick control. Additional barriers to implementing community-based tick management programs within residential communities are discussed.

Selective versus strategic control against Rhipicephalus microplus in cattle: A comparative analysis of efficacy, animal health, productivity, cost, and resistance management.

This study compared selective control versus strategic control against Rhipicephalus (Boophilus) microplus, under conditions of a real dairy farm. Strategic control requires that all animals be treated with acaricide at regular pre-determined intervals. Selective control only requires treatment of infested animals and only when they are at or above a pre-determined threshold. Tick counts on animals and in pasture were performed and the susceptibility of tick populations to the different treatment methods was evaluated at the beginning and end of the study using the Larval Packet Test, Larval Immersion Test, and Adult Immersion Test. Over the four years of the study strategic control was more advantageous than the selective control as the group experienced lower tick burden on animals and in pasture, absence of skin lesions and myiases, lower operating costs, treatments concentrated in fewer months of the year and lower resistance pressure.

An economic evaluation of cattle tick acaricide-resistances and the financial losses in subtropical dairy farms of Ecuador: A farm system approach.

Estimates of economic losses in cattle due to tick infestations in subtropical areas are limited, such as in Ecuador. Ticks affect animal production and health, but those direct effects are difficult to estimate since financial exercises carried out in farms consider both costs of the inputs and revenues. This study aims to quantify the costs of inputs involved in milk production and to know the role of acaricide treatment in the production costs on dairy farms in subtropical zones using a farming system approach. Regression and classification trees were used to study the relationship between tick control, acaricide resistance and the presence of high level of tick infestation in the farm system. Even though there was no significant direct association between high levels of tick infestation and the presence of acaricide resistance in ticks, a more complex structure for resistances operates in the manifestation of high tick infestation involving levels of farm technology and no acaricide resistance. Farms with higher levels of technology allocate a lower percentage of sanitary expenses to control ticks (13.41%) in comparison to semi-technified (23.97%) and non-technified farms (32.49%). Likewise, more technified and bigger herds have a lower annual expenditure on acaricide treatment (1.30% of the production budget equivalent to 8.46 USD per animal) compared to non-technified farms where it can represent more than 2.74% of the production budget and where the absence of cypermethrin resistance increases the treatment cost to 19.50 USS per animal annually. These results can motivate the development of information campaigns and control programmes targeted to the reality of small and medium farms that are the most affected in terms of the money they invest in controlling ticks.

Assessment of cypermethrin and amitraz resistance and molecular profiling of voltage-gated sodium channel and octopamine tyramine genes of Rhipicephalus microplus.

Control of ticks and tick-borne pathogens is a priority for human and animal health. Livestock-holders extensively rely on acaricide applications for tick control. Different groups of acaricides including cypermethrin and amitraz have been consistently used in Pakistan. There has been a gap in understanding the susceptibility or resistance of Rhipicephalus microplus, the most prevalent tick in Pakistan, to acaricides. The present study aimed to molecularly characterize cypermethrin and amitraz targeted genes such as voltage-gated sodium channel (VGSC) and octopamine tyramine (OCT/Tyr) of R. microplus ticks in Khyber Pakhtunkhwa (KP), Pakistan to monitor the acaricides resistance. Tick specimens were collected from cattle and buffaloes in northern (Chitral, Shangla, Swat, Dir, and Buner), central (Peshawar, Mardan, Charsadda, Swabi, and Nowshera), and southern districts (Kohat, Karak, Lakki Marwat, Tank, and Dera Ismail Khan) of KP, Pakistan. Different concentrations of commercially available cypermethrin (10%) and amitraz (12.5%) were prepared for in vitro larval immersion tests (LIT). In LIT, the average mortality rate of immersed larvae was recorded that was increased gradually with an increase in the concentration of specific acaricide. The larvae's highest mortality rates (94.5% and 79.5%) were observed at 100-ppm of cypermethrin and amitraz, respectively. A subset of 82 R. microplus ticks was subjected to extract genomic DNA, followed by PCR to amplify partial fragments of VGSC (domain-II) and OCT/Tyr genes. The BLAST results of the consensus sequence of VGSC gene (domain-II) showed 100% identity with the acaricides susceptible tick sequence from the United States (reference sequence). Obtained identical sequences of OCT/Tyr genes showed maximum identity (94-100%) with the identical sequences reported from Australia (reference sequence), India, Brazil, Philippines, USA, South Africa, and China. Thirteen single nucleotide polymorphisms (10 synonymous and three non-synonymous) were observed at various positions of partial OCT/Tyr gene fragments. The SNP at position A-22-C (T-8-P) in OCT/Tyr gene has been linked to amitraz resistance in R. microplus ticks. Molecular analysis and LIT bioassay's findings indicate the availability of resistant R. microplus ticks in the KP region. To our understanding, this is the first preliminary study to monitor cypermethrin and amitraz resistance via molecular profiling of cypermethrin and amitraz targeted genes (VGSC and OCT/Tyr) in combination with in vitro bioassays (LIT) in R. microplus ticks from Pakistan.

Acaricidal efficacy evaluation of amitraz and diazinon against Amblyomma variegatum tick species in Waghimra zone, northern Ethiopia.

Ticks pose a substantial economic burden associated with production loss and treatment costs globally. Ethiopia has tremendous livestock resources, but its productivity is hindered by various animal health challenges in which ticks are the top priority, with a limited response to acaricidal treatments. Hence, we designed an acaricidal efficacy trial for the top commercially available Chemicals (amitraz and diazinon) to examine their efficacy against the widely distributed tick species (Amblyoma variegatum). Ticks were collected from animals admitted to veterinary clinics with no history of treatment with acaricides. Adult Immersion Technique (AIT) was employed to detect acaricidal resistance, and the mean percent control and antiparasitic efficacy were used to estimate tick susceptibility with a completely randomized laboratory-based trial (CRT). The mean weights of eggs laid by ticks subjected to amitraz and diazinon indicated that amitraz had a better egg-laying-inhibition effect than diazinon. The mean control percentages of amitraz and diazinon were 92.8 ± 5.6% and 69.7 ± 3.1%, respectively, with a highly significant difference (P-value = 0.00). The antiparasitic efficacy of the two drugs was 57.5 ± 0.96 and 37.5% ± 0.96 for amitraz and diazinon, respectively, which revealed that amitraz was statistically better than diazinon in killing adult ticks (P-value =0.026). In general, ticks treated with diazinon showed evidence of resistance development, and amitraz is relatively the most effective acaricide; we recommend its use in the study area and other locations with similar settings.

Toxicity assessment of common acaricides and mineral oils on Anagyrus vladimiri, an effective biocontrol agent of citrus mealybug.

Chemical pesticides, while playing an important role in the suppression of insect pests, should be used in a manner that minimizes negative effects on natural enemies. The parasitoid, Anagyrus vladimiri Triapitsyn (Hymenoptera: Encyrtidae), plays an important role in the management of mealybug pests of citrus groves in the Mediterranean region. This study was conducted to evaluate the effect of commonly used acaricides (Spirodiclofen, Spirotetramat, Sulfur, Fenpyroximate, Abamectin) and mineral oils (Levanola, EOS, JMS, and Ultrapaz) on acute mortality of A. vladimiri. Toxicity was assessed in 4 cases: (i) direct spray application on adults, (ii) pesticide application on the mummified host, (iii) feeding with contaminated food, and (iv) contact with pesticide residue. The pesticide Abamectin, applied alone and with Levanola oil was highly toxic to adults in all bioassays, with the exception of direct spray application on the mummified host. Fenpyroximate was found to be highly toxic only when sprayed directly on adults, and sulfur was slightly harmful. Mineral oils were harmful when ingested with food; otherwise, they did not cause appreciable adult mortality. The findings of the present study suggest that all tested materials, with the exception of Abamectin and Fenpyroximate, are compatible with the survival of A. vladimiri. Direct ingestion of oils can, however, cause a degree of mortality. Given that indiscriminate use of these pesticides may affect the population ecology of A. vladimiri, they should be used with caution.

Resistance of Panonychus citri (McGregor) (Acari: Tetranychidae) to pyridaben in China: monitoring and fitness costs.

BACKGROUND: Panonychus citri is a major citrus pest worldwide. The short life cycle and high reproductive potential of P. citri, combined with heavy acaricide use, have led to high levels of resistance to acaricides, posing a threat to global resistance management programs. Here, resistance monitoring was established to determine the pyridaben resistance status of ten P. citri populations in China from 2014 to 2021 using a leaf-dipping assay. Four characterized strains-the susceptible strain (Lab_S), the resistant strain (Pyr_R), as well as the segregated resistant strain (Pyr_Rs) and the segregated susceptible strain (Pyr_Control) derived from the crossing of the Lab_S and Pyr_R strains, were used to evaluate the life-history characteristics using age-stage, two-sex life tables. RESULTS: Most P. citri populations developed high resistance to pyridaben. Resistance levels exceeded 1000-fold in Yuxi, Anyue, Nanning, and Ganzhou populations compared with the Lab_S strain. Compared with Pyr_Control, two key fitness cost criteria, developmental period and fecundity, showed significant differences in Pyr_Rs under consistent conditions. The intrinsic rate of increase, net reproductive rate and gross reproductive rate were lower in the resistant strain compared with the Pyr_Control strain. The Pyr_Rs strain had a lower relative fitness of 0.934 compared with the Pyr_Control. Moreover, the life-history traits and population parameters of the Pyr_R strain also showed significant differences compared with the Lab_S strain. CONCLUSION: The resistance levels to pyridaben varied greatly among the different P. citri populations and showed regional differences. Substantial fitness costs are associated with pyridaben resistance. This study provides potential implications for developing strategies for resistance management in P. citri. © 2022 Society of Chemical Industry.

Economic impact of predominant ticks and tick-borne diseases on Indian dairy production systems.

As per 20th livestock census, India possessing 193.46 million cattle and 109.85 million buffaloes in organized and unorganized sectors. These animals are suffering from tick infestation almost throughout the year causing both direct and indirect losses. Repeated use of chemical acaricides for tick management resulted in the establishment of acaricide resistant tick populations, insecticide residues in livestock products and environmental pollution. In the present study, analysis of the data generated during 1990-2021 revealed pooled prevalence of infestations in cattle and buffaloes was 53% [95% CI = 47-60%] and 38% [95% CI = 28-49%], respectively. Based on peripheral blood smear examination the prevalence of theileriosis, babesiosis and anaplasmosis in cattle is estimated as 17% [95% CI = 12-24%], 4% [95% CI = 3-6%] and 5% [95% CI = 3-7%], respectively, while in buffalo the prevalence of infection is estimated as 7% [95% CI = 2-21%], 2% [95% CI = 0-5%] and 8% [95% CI = 2-36%],respectively.To estimate economic impact, both direct (reduction in milk production, cost of treatment, leather depreciation) and indirect (milk loss and treatment cost) losses were taken into consideration. Loss of milk production was predicted as 13.91, 56.91 and 85.34L/cross-bred cow/lactation in low, moderate and high tick infestation conditions, respectively. Whereas, 20.10, 7.01L milk/buffalo/lactation in Hyalomma spp. and Rhipicephalus spp. infestation was estimated. Similarly,the estimated loss of milk production due to clinical theileriosis, babesiosis and anaplasmosis was 57.96, 30.96 and 59.22L, respectively. The cumulative (milk loss, treatment cost and leather loss) loss due to tick infestation was calculated as 46199.31 million INR (USD595.07 million) while due to TBDs 14877.15 million INR (USD191.15 million) = 61076.46 million INR (USD787.63 million). The data provided base line information for the policy maker to develop strategies at government level so that the significantly high cumulative loss of 787.63 million USD due to ticks and tick borne diseases (TTBDs) can be minimized.

Structure-based in silico design and in vitro acaricidal activity assessment of Acacia nilotica and Psidium guajava extracts against Sarcoptes scabiei var. cuniculi.

Infestation by Sarcoptes scabiei var. cuniculi mite causes scabies in humans and mange in animals. Alternative methods for developing environmentally friendly and effective plant-based acaricides are now a priority. The purpose of this research was the in silico design and in vitro evaluation of the efficacy of ethanol extracts of Acacia nilotica and Psidium guajava plant leaves against S. scabiei. Chem-Draw ultra-software (v. 12.0.2.1076.2010) was used to draw 36 distinct compounds from these plants that were employed as ligands in docking tests against S. scabiei Aspartic protease (SsAP). With docking scores of - 6.50993 and - 6.16359, respectively, clionasterol (PubChem CID 457801) and mangiferin (PubChem CID 5281647) from A. nilotica inhibited the targeted protein SsAP, while only beta-sitosterol (PubChem CID 222284) from P. guajava interacted with the SsAP active site with a docking score of - 6.20532. Mortality in contact bioassay at concentrations of 0.25, 0.5, 1.0, and 2.0 g/ml was determined to calculate median lethal time (LT50) and median lethal concentration (LC50) values. Acacia nilotica extract had an LC50 value of 0.218 g/ml compared to P. guajava extract, which had an LC50 value of 0.829 g/ml at 6 h. These results suggest that A. nilotica extract is more effective in killing mites, and these plants may have novel acaricidal properties against S. scabiei. Further research should focus on A. nilotica as a potential substitute for clinically available acaricides against resistant mites.

Control Strategies for the Tick Rhipicephalus Microplus (Canestrini, 1888) on Cattle: Economic Evaluation and Report of a Multidrug-Resistant Strain.

PURPOSE: The aim of this study was to evaluate four strategies for application of acaricides to control the tick Rhipicephalus microplus among infested cattle, and to show which of these has the best cost-benefit ratio. METHODS: For this, 72 cattle were selected and divided into four groups: Group 1 (G1): fipronil, pour-on; Group 2 (G2): fluazuron, pour-on; Group 3 (G3): moxidectin, injectable; and Group 4 (G4): chlorpyrifos 30 g, cypermethrin 15 g and fenthion 15 g, spraying (atomizing chamber). Every seven days, the numbers of semi-engorged females were counted and laboratory tests were conducted using different commercial technical-grade products for resistance monitoring. RESULTS: G4 showed the best percentage reduction, with the highest rate on the seventh day post-treatment (DPT) (83.23%). G3 was the second best strategy, with a percentage of inverse reduction such that the best results were on the 28th DPT (82.85%), while G1 and G2 reached their best results on the 21st DPT (32.63% and 2.79%). CONCLUSION: It was noteworthy that the formulation used in G4 was the only one that was efficient for strategic control and that, based on the economic analysis, was shown to be economically viable over the medium term due to the need for investment. The presence of a multidrug-resistant strain in the state of Mato Grosso do Sul in vitro, for the chemical bases amitraz, cypermethrin and cypermethrin + DDVP, is reported here for the first time.

In vitro assessment of the acaricidal activity of a carvacrol shampoo on tick larvae.

Ticks are a widely distributed arthropod of veterinary importance. Resistance of ticks to synthetic acaricides has become widespread, warranting the development of new drugs for tick management. Carvacrol is a volatile monoterpene, with promising results against various species of ticks; however, to be used for therapeutic purposes, carvacrol must be included in a formulation that makes its application feasible. This study aims to develop a formulation of a carvacrol-containing shampoo that is effective against two species of ticks: Rhipicephalus sanguineus and R. microplus. Shampoo sensory characteristics and pH were evaluated at 37, 25 and 5 °C, for a maximum of 15 days. The shampoo remained stable at 25 and 5 °C. The efficacy of the carvacrol-containing formulation against two species of ticks was assessed by the larval immersion test. Mortality of both tick species was significantly higher for the carvacrol shampoo than for a carvacrol hydroalcoholic solution. In conclusion, the carvacrol-containing shampoo showed larvicidal efficacy on ticks.

Lithium salts: assessment of their chronic and acute toxicities to honey bees and their anti-Varroa field efficacy.

BACKGROUND: Varroa control is essential for the maintenance of healthy honey bee colonies. Overuse of acaricides has led to the evolution of resistance to those substances. Studies of the short-term acaricidal effects and safety of various lithium (Li) salts recently have been reported. This study examined the long-term in vitro and in vivo bee toxicities, short-term motor toxicity to bees and long-term anti-Varroa field efficacy of several Li salts. RESULTS: In an in vitro chronic-toxicity assay, lithium citrate (18.8 mm) was the most toxic of the examined salts, followed by lithium lactate (29.5 mm), and lithium formate (32.5 mm). In terms of acute locomotor toxicity to bees, all of the Li salts were well-tolerated and none of the treatment groups differed from the negative control group. In an in vitro survival study, all of the Li treatments significantly reduced bee life spans by a factor of 1.8-7.2, as compared to the control. In terms of life expectancy, lithium citrate was the most toxic salt, with no significant differences noted between lithium formate and lithium lactate. In the bee-mortality field study, none of the examined treatments differed from the negative control. Amitraz and lithium formate exhibited similar acaricide effects, which were significantly different from those observed for lithium lactate and the negative control. CONCLUSION: In light of lithium formate's honey bee safety and efficacy as an acaricide, additional sublethal toxicity studies in brood, drones and queens, as well as tests aimed at the optimization of administration frequency are warranted. © 2022 Society of Chemical Industry.

Pyrokinin receptor silencing in females of the southern cattle tick Rhipicephalus (Boophilus) microplus is associated with a reproductive fitness cost.

BACKGROUND: Rhipicephalus microplus is the vector of deadly cattle pathogens, especially Babesia spp., for which a recombinant vaccine is not available. Therefore, disease control depends on tick vector control. However, R. microplus populations worldwide have developed resistance to available acaricides, prompting the search for novel acaricide targets. G protein-coupled receptors (GPCRs) are involved in the regulation of many physiological processes and have been suggested as druggable targets for the control of arthropod vectors. Arthropod-specific signaling systems of small neuropeptides are being investigated for this purpose. The pyrokinin receptor (PKR) is a GPCR previously characterized in ticks. Myotropic activity of pyrokinins in feeding-related tissues of Rhipicephalus sanguineus and Ixodes scapularis was recently reported. METHODS: The R. microplus pyrokinin receptor (Rhimi-PKR) was silenced through RNA interference (RNAi) in female ticks. To optimize RNAi, a dual-luciferase assay was applied to determine the silencing efficiency of two Rhimi-PKR double-stranded RNAs (dsRNA) prior to injecting dsRNA in ticks to be placed on cattle. Phenotypic variables of female ticks obtained at the endpoint of the RNAi experiment were compared to those of control female ticks (non-injected and beta-lactamase dsRNA-injected). Rhimi-PKR silencing was verified by quantitative reverse-transcriptase PCR in whole females and dissected tissues. RESULTS: The Rhimi-PKR transcript was expressed in all developmental stages. Rhimi-PKR silencing was confirmed in whole ticks 4 days after injection, and in the tick carcass, ovary and synganglion 6 days after injection. Rhimi-PKR silencing was associated with an increased mortality and decreased weight of both surviving females and egg masses (P < 0.05). Delays in repletion, pre-oviposition and incubation periods were observed (P < 0.05). CONCLUSIONS: Rhimi-PKR silencing negatively affected female reproductive fitness. The PKR appears to be directly or indirectly associated with the regulation of female feeding and/or reproductive output in R. microplus. Antagonists of the pyrokinin signaling system could be explored for tick control.

Field efficacy assessment of a vaccine against Rhipicephalus (Boophilus) australis in New-Caledonia.

Pure European bovine breeding is compromised in South Pacific (i.e. Australia, New Caledonia) due to the impact of acaricide-resistant Rhipicephalus (Boophilus) australis ticks. Farmers need new solutions to sustainably maintain their activity. An integrated tick control program has been applied for 2 years in herds of European breeds (i.e. Charolais and Limousine cattle breeds) in New Caledonia. A field trial including the use of a vaccine based on a local Bm86 antigen, agropastoral measures and a chemical control of the tick was conducted to assess the possibility to reduce the number of acaricidal treatments. Nine farms including 1400 animals were monitored from July 2017 to June 2020. The year mid2017-mid2018 was the year of initial monitoring in order to get control data related to tick infestation during the seasons and to the number of treatments per herd. Vaccinations began between September and December 2018. Results obtained from mid2019 to mid2020 were then compared to the initial period (mid2017-mid2018) to assess the benefits of the vaccine. The average annual number of acaricide treatments decreased significantly from 7.44 before vaccination to 4.45 during mid2019-mid2020 (p = 0.026). Infestation scores, assessed by tick counts when farmers grouped the cattle to apply acaricides, showed a constant and low infestation level. Nevertheless, farmers were willing to tolerate slightly higher tick loads on vaccinated cattle with less acaricidal treatments compared to unvaccinated and frequently treated cattle. Local cutaneous reactions were recorded at the injection site with 46.6% of all injections evolving into nodules and 1% of injections giving depilation or abscesses. At the end of the trial, all the breeders wish to continue the vaccination and consider that it is now possible to serenely maintain their breed.

The bioactive compound carvacrol as a potential acaricide: An assessment of its effects on the integument of female Rhipicephalus sanguineus sensu lato ticks.

Studies seeking control methods for infestation of Riphicephalus sanguineus sensu lato (s.l.) ticks (dog ticks) have been carried out in order to minimize damage to both the tick's hosts and the environment, the latter due to the misuse of acaricide products. In this regard, carvacrol has been used as a natural alternative against ticks as it displays several properties including acaricidal. In this context, the present study aimed to evaluate the ultramorphology and morphohistochemistry of the integument of semi-engorged R. sanguineus s.l. females exposed to different carvacrol concentrations. The findings indicate that the integument surface of females exposed to the highest carvacrol concentration (25 µl/ml) became wrinkled, suggesting dehydration or the result of integument cuticular and epithelial layer disorganization in response to the toxic product. Morphohistochemical integument layer alterations were more significant and intense in females exposed to the highest carvacrol concentration (25 µl/ml), confirming dose-dependent carvacrol action. Among other cell and tissue alterations, changes in epithelial cell shape, size, and arrangement (epidermal layer) were noted, alongside altered and pyknotic-shaped nuclei, suggesting a death process for these cells. This epithelium changed from simple cubic to stratified, also in response to the presence of the evaluated bioactive compound. Thus, the findings reported herein demonstrate that carvacrol may be an alternative for an efficient and more sustainable tick control in the near future.

Successive treatments with ivermectin (3.15%) to control the tick Rhipicephalus (Boophilus) microplus in cattle: Pharmacokinetic and efficacy assessment.

This study aimed to evaluate the pharmacokinetics, the potential accumulation in the body of treated animals and the efficacy of ivermectin long-acting formulation (3.15%) against the cattle tick Rhipicephalus (Boophilus) microplus in a scheme of three successive treatments. Fifteen 12-month-old heifers, naturally infested with R. microplus, were divided into two groups (G). Cattle from GI (n = 10) were subjected to three treatments with ivermectin 3.15% (IVOMEC GOLD®, Merial Argentina S.A.) at a rate of 1 mL/50 kg on days 0, 35, and 70. Cattle from GII (n = 5) were not treated. From day 1 to 202 post-treatment blood samples were taken to measure ivermectin concentrations by HPLC and female ticks (4.5-8 mm) were counted to evaluate the efficacy of the treatment. The level of tick resistance to ivermectin was evaluated before and after finishing the scheme of successive treatments by larval immersion test (LIT) bioassay from engorged females collected from GI. The area under the concentration vs. time curves (AUC0-35d) obtained post-second treatment was 1.51 ± 0.39-fold higher than those observed post-first treatment (P<0.05). The mean plasma concentrations of ivermectin 3.15% at 20 days after the first, second and third treatment were 17.0, 27.5 and 37.8 ng/mL, respectively (P<0.01). The elimination half-life of ivermectin post-third treatment was significantly longer than that was previously reported after a single dose (P<0.01). Values of therapeutic efficacy percentage reached 75.6% post-first treatment and between 95.9 and 100% after the second treatment. Ticks evaluated by LIT showed a significant increase in lethal concentrations after treatments. Although the efficacy level was high, the successive treatments with long-acting ivermectin formulation generate a significant accumulation of drug in plasma and could increase the levels of resistance to this drug in the tick population.