Essential oil pharmaceuticals for killing ectoparasites on dogs.

BACKGROUND: External parasites, particularly ticks and fleas, are among the most common problems affecting dogs. Chemical medicines are commonly used to prevent and eliminate such external parasites, but their improper use can cause adverse reactions, and the toxins they contain may remain in the environment. OBJECTIVES: The objective of this study was to investigate the in vitro efficacy of Zanthoxylum limonella, citronella, clove, peppermint, and ginger essential oils against dog ticks and fleas and to test the sensitivity of dogs' skin to these essential oils. METHODS: The five essential oils were tested for in vitro efficacy against ticks and fleas, and the two most effective essential oils were then tested on the dogs' skin. RESULTS: The results revealed that these five essential oils at 16% concentrations effectively inhibited the spawning of female engorged ticks. In addition, all five essential oils had a strong ability to kill tick larvae at concentrations of 2% upward. Furthermore, 4% concentrations of the five essential oils quickly eliminated fleas, especially clove oil, which killed 100% of fleas within 1 h. A 50%, 90%, and 99% lethal concentration (LC50, LC90, and LC99) for the essential oils on tick larvae in 24 h were found to be low values. LC50, LC90, and LC99 for the essential oils on flea in 1 h was lowest values. Clove oil at 16% concentration was the most satisfactory essential oil for application on dogs' skin, with a low percentage of adverse effects. CONCLUSIONS: This study confirmed the effectiveness of essential oils for practical use as tick and flea repellents and eliminators. Essential-oil-based pharmaceutical can replace chemical pesticides and provide benefits for both consumers and the environment.

Evaluation of synergism in essential oils against the cattle tick Rhipicephalus microplus in Burkina Faso.

The cattle tick Rhipicephalus microplus affects animal production economically by reducing weight gain and milk production and causing diseases, such as babesiosis and anaplasmosis. Using synthetic acaricides to reduce their incidence has caused the emergence of resistant tick populations. The present study aimed to assess the in vitro acaricidal activity of combinations of essential oils (EOs) from Ocimum americanum, Ocimum gratissimum, and Lippia multiflora against R. microplus larvae. In fact, numerous biological properties have been reported on EOs from these three plants, including acaricidal properties. Hence, a larval immersion test was performed using a population of R. microplus resistant to synthetic acaricides used in Burkina Faso. Results revealed that EO from O. gratissimum was the most effective on R. microplus larvae with LC50 and LC90 values at 10.36 and 15.51 mg/mL, respectively. For EO combinations, the most significant synergistic effect was obtained by combination 6 (1/3 O. americanum + 2/3 O. gratissimum +1/6 L. multiflora), with a combination index value of 0.44. All combinations presented dose reduction index >1, indicating a favorable dose reduction. According to the literature, this is the first study to determine the combination effect of EOs from the abovementioned plants in controlling R. microplus activity in vitro. Thus, the combination of these EOs is an alternative to control the resistant populations of invasive cattle ticks.

Efficacy of hydroethanolic extract of Randia aculeata seed against the southern cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) on naturally infested cattle under field conditions.

The cattle tick Rhipicephalus (Boophilus) microplus is a major problem of concern for cattle industry in tropical and subtropical areas. Control of cattle tick is based mainly on the use of chemical acaricides, which has contributed to the emerging problem of selection of resistant tick lineages. Plants have been used as an alternative to conventional acaricidal drugs. On the other hand, the acaricidal activity of hydroethanolic extract of Randia aculeata seed (EHRA) has been demonstrated against R. microplus under laboratory conditions. However, the utility of EHRA seed as a potential acaricidal needs to be determined under field conditions. For this reason, the aim of this study was to evaluate the efficacy of the EHRA against R. microplus sprayed on naturally infested calves, determine the effect of the EHRA seed on acetylcholinesterase activity in R. microplus larval and identify the chemical composition of EHRA. Forty-five male calves were divided in three groups and treated with: G1 water; G2 EHRA 20% w/v and G3 coumaphos 0.2% v/v. Acetylcholinesterase (AChE) activity in R. microplus larvae was determined by a colorimetric assay. The chemical composition of EHRA was accessed through HPLC/MS. Significantly fewer ticks were observed after 24 h on the treated group compared to control group. EHRA significantly inhibited in vitro AChE activity in R. microplus at all tested concentrations. Chlorogenic acid, vanillinic acid, p-coumaric acid, caffeic acid. rutin, quercetin, (-)-epicatechin, 4-hydroxybenzoic acid, quercetin, vanillin, 2,4-dimethoxy-6-methylbenzoic acid, scopoletin and ferulic acid were identified in the extract. The results provided new data for the elucidation of the mechanisms of EHRA acaricide action and to further evaluate the use as a new alternative control agent against R. microplus under in vivo conditions.

Establishment of antitick efficacy of a phytoformulation prepared from Annona squamosa leaf extracts for the management of acaricide resistant tick infestations on cattle.

To combat the problem of acaricide resistance in ticks, efforts have been made to develop eco-friendly herbal acaricides which are safe for animal use with no possibilities of evoking environment and residual toxicity. In the present study, the acaricidal properties of a commonly available plant, Annona squamosa have been established to develop a novel antitick phytoformulation using leaf extract, suitable solvents and emulsifier for managing resistant tick infestations in animals. The quality control of the safe and stable phytoformulation using Rutin as a major constituent was maintained by HPTLC profiling. Several phytformulations (A-E) were prepared and formulation 'C' is identifried as the most effective showing 52.5-75% antitick activity against in vitro treated resistant strains of R. microplus (IVRI-IV and V) with 33.8-40.2% inhibition of oviposition while 65-85% mortality against multiacaricide resistant field populations. The phytoformulation provided 70.4% efficacy after first larval challenge in experimentally infested animals. The field efficacy of the formulation in different multilocational field trials was 68.53-77.8%. The detail antitick efficacy of the formulation along with safety and stability are explained as a component of antitck technology to manage resistant tick problem in livestock.

Acaricidal and repellent activity of the essential oils of Backhousia citriodora, Callistemon viminalis and Cinnamodendron dinisii against Rhipicephalus spp.

The ticks Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato (s.l) are of great importance to agriculture, veterinary medicine and public health. Due to a number of problems related to the use of synthetic acaricides, natural products emerge as promising substances for alternative tick control. In the present study, essential oils of Backhousia citriodora, Callistemon viminalis and Cinnamodendron dinisii were extracted by hydrodistillation, characterized by GC-MS and GC-FID and biologically evaluated for acaricidal activity against R. microplus and repellent activity against R. sanguineus s.l. Inhibition of acetylcholinesterase (AChe) by the essential oils was also evaluated. The major constituent of B. citriodora is citral (98.9 %), whereas the essential oil from C. viminalis is rich in 1.8-cineole (78.1 %), α-pinene (12.5 %) and limonene (3.36 %), and that from C. dinisii contains α-pinene (30.8 %), ß-pinene (12.5 %) and sabinene (11.3 %) as the principal constituents. The median lethal concentrations (LC 50) estimated for the essential oils on engorged R. microplus females were 3.276 µL.mL-1 for B. citriodora, 8.195 µL.mL-1 for C. dinisii and 8.936 µL.mL-1 for C. viminalis. The essential oil of B. citriodora showed the best repellent effect against unfed R. sanguineus s.l. adults, demonstrating repellent action up to 3 h after application. The essential oil of C. viminalis was able to reduce AChe activity, with an average inhibitory concentration (IC 50) of 0.33 µg mL-1. Thus, these oils can be considered as sources of bioactive compounds for tick control.

First report on efficacy of Citrus limetta seed oil in controlling cattle tick Rhipicephalus microplus in red Sahiwal calves.

The objective of the study was to evaluate the acaricidal activity of Citrus limetta seed oil (CLO) for controlling the cattle tick Rhipicephalus microplus. C. limetta seeds were collected as a waste product from different juice corners. CLO was obtained after extraction of seeds on soxhlet apparatus using n-hexane as solvent. It was characterized through Gas Chromatography-High Resolution Mass Spectroscopy (GC-HRMS) to determine the presence of active constituents. In vitro bioassays were performed using adult immersion test (AIT) and larval packet test (LPT). In vivo acaricidal efficacy of CLO was performed on red Sahiwal calves using ear bag method. Clinical safety of CLO was evaluated by observing haematological parameters and skin irritancy assay. Results of GC-HRMS showed that mainly fatty acids such as linoleic acid, stearic acid, oleic acid, palmitic acid, and linolenic acid were present in the CLO. CLO in the concentration of 125 mg/mL (CLO8) exhibited 100 % mortality in both AIT and LPT. CLO significantly (p < 0.001) reduced the number of ticks from 35 to 5.05 and 3.24 on 144 h after treatment with CLO7 and CLO8, respectively. CLO was found clinically safe without producing erythema and edema on skin. Haematological parameters such as haemoglobin (11.48 g/100 mL), total leucocytes count (4.32 106/cumm), total erythrocytes count (6.80 106/cumm), and packed cell volume (34.39 %) were normal and controlled. CLO may be used as effective and safe drug therapy for controlling R. microplus ticks.

Evaluation on two types of paramyosin vaccines for the control of Haemaphysalis longicornis infestations in rabbits.

BACKGROUND: Haemaphysalis longicornis is an obligate hematophagous ectoparasite that transmits a variety of pathogens causing life-threatening diseases in humans and animals. Paramyosin (Pmy) is not only an invertebrate-specific myofibrillar protein but also an important immunomodulatory protein. Therefore, it is one of the ideal candidate antigens for vaccines. METHODS: We conducted two vaccine trials to evaluate the protective efficacy of Pmy recombinant protein (rPmy) and peptide vaccine (KLH-LEE). Each rabbit was immunized with three doses of rPmy or KLH-LEE adjuvanted with Freund's complete/incomplete at 500 µg/dose at 2-week intervals before challenge with 40 female H. longicornis/rabbit. PBS plus adjuvant, Trx or KLH was used as control group. The antibodies of rabbits were detected by ELISA. Then, female ticks were fed on the rabbits until detachment. RESULTS: ELISA results showed that both vaccines induced rabbits to produce antibodies. Compared with the Trx group, the engorgement weight, oviposition and hatchability of the rPmy group decreased by 8.87%, 26.83% and 38.86%, respectively. On the other hand, engorgement weight, oviposition and hatchability of female ticks in the KLH-LEE group correspondingly resulted in 27.03%, 53.15% and 38.40% reduction compared with that of the KLH group. Considering the cumulative effect of vaccination on the evaluated parameters, results showed 60.37% efficacy of the rPmy vaccine formulation and 70.86% efficacy in the KLH-LEE group. CONCLUSIONS: Pmy and particularly epitope LEE have potential for further development of an effective candidate vaccine to protect the host against tick infection. GRAPHIC ABSTARCT.

Acaricidal efficacy and chemical study of hexane extracts of the leaves of Neoglaziovia variegata (Bromeliaceae) against the tick Rhipicephalus microplus.

The frequent use of acaricides against the tick Rhipicephalus microplus increases the risk of development of resistance. Recent studies have revealed that Neoglaziovia variegata, an indigenous plant species known in Brazil as 'caroá', has a deleterious effect against R. microplus. In the current study, extracts of N. variegata were studied for their possible acaricidal properties. A hexane extract of N. variegata leaves was fractionated in a chromatography column and the fractions were tested in adult tick immersion tests in triplicate using three concentrations (5, 10 and 25 mg/ml). All the fractions had harmful effects on the ticks. However, three fractions were more efficaceous. Phytochemical analysis indicated that stigmast-5-en-3-ol and stigmastanol were most abundant; they might be responsible for the acaricidal effects, making them potentially useful as alternative agents to control the tick R. microplus.

Development of an efficient antitick natural formulation for the control of acaricide-resistant ticks on livestock.

Animal production has a key role in global economic development and food security. Ticks, specifically Rhipicephalus microplus cause substantial economic and health impacts on more than eighty percent of the world cattle population. Though synthetic acaricides play a major role in tick management, their injudicious usage has caused environmental pollution and also promote the establishment of multi-acaricide resistant tick populations which is a matter of great concern. To provide an effective tool for controlling these resistant ticks, the present work was aimed to develop safe and inexpensive antitick natural formulations. Our bioprospection studies of Ageratum conyzoides plant established it as a species potentially having strong acaricidal activity due to the presence of potent acaricidal phyto-chemicals. To develop a suitable antitick natural formulation, 41 samples/fractions/formulations were prepared from the dry powder of the whole aerial part of the A. conyzoides plant using different techniques and delivery matrices. The strongest antitick effect was recorded for formulation ACF6, which demonstrated 87 ± 6% mean mortality with 57 % inhibition of oviposition in treated female ticks. Ticks treated with the ACF6 formulation showed a significant (p < 0.001) reduction in cuticular protein (1.238 ± 0.01 mg/mL) as compared to control ticks (2.928 ± 0.01 mg/mL) but no significant difference in chitin content of treated ticks and control ticks was observed. The formulation was found safe in a rat model as no significant differences in biochemical and haematological parameters among treated and control rats were noted. Histopathological studies indicated no sign of hepatocellular necrosis and no significant changes in the weights of liver and spleen was recorded. The overall in vivo efficacy of the formulation was 85 % for experimentally infested cattle with direct mortality of more than 80 % within 96 h post-application. The lethal effect of the formulation was in the form of drying and dead ticks 1-2 d after application. The developed formulation has the potential to be adopted as an alternative tick control measure in an ecofriendly manner.

Control of Rhipicephalus annulatus resistant to deltamethrin by spraying infested cattle with synergistic eucalyptus essential oil-thymol-deltamethrin combination.

The current study investigated the synergistic effect of combinations containing deltamethrin (D), Eucalyptus essential oil (E), and the thyme essential oil component thymol (T), against a field population of Rhipicephalus annulatus in Egypt that was characterized to be resistant to D. Solutions of T, E, or TE at concentrations of 1.25-5% were combined with 5% deltamethrin at different dilutions (0.25-2 mL/L). Results of the adult immersion test used to estimate the in vitro acaricidal activity of these combinations at 5% yielded LC50 values for D, E-D, T-D, and TE-D of 3.87 mL/L, 3.89 mL/L, 0.14 mL/L, and 0.05 mL/L, respectively. Biochemical analyses using whole-body homogenate of ticks from the in vitro tests revealed that the lowest acetylcholinesterase and glutathione peroxidase activity, and the maximum lipid peroxidation were recorded in ticks treated with 5% TE-D. Glutathione content significantly decreased (p ≤ 0.05) in all treated ticks. Three groups, each containing five cross breed cattle naturally infested with R. annulatus from the same area where resistance to D was detected, were sprayed twice at two-week intervals using 1 mL/L of 5% solutions of D, T-D, or TE-D. Overall efficacy of the D, T-D, and TE-D sprays by day 30 post-treatment was 21.6, 88.3, and 95 %, respectively. Ticks collected from infested cattle three days after treatment with the D spray deposited egg masses that were able to hatch, deposited small masses of eggs unable to hatch when exposed to the T-D spray, and laid few eggs that didn't hatch when sprayed with the TE-D combination. Values for liver and kidney function parameters were comparable in cattle before and after treatment with the combination sprays tested. The TE-D spray overcame the insensitivity to D of this R. annulatus population in Egypt, which also highlighted the significant synergistic effect of thymol on the acaricidal activity of deltamethrin observed in vitro. Acaricidal activity of the TE-D combination apparently has deleterious effects on multiple tick systems involving inhibition of acetylcholinesterase, increased lipid peroxidation, and oxidative stress. These findings document that combinations of natural and synthetic products can be part of integrated management solutions to the problem with widespread resistance to pyrethroids like deltamethrin in populations of cattle ticks, including R. annulatus, around the world.

Rhipicephalus microplus (acari: Ixodidae): Clinical safety and potential control by topical application of cottonseed oil (Gossypium sp.) on cattle.

The present study was performed to determine the acaricidal activity of the cottonseed oil (CSO) against cattle tick Rhipicephalus microplus. CSO was analyzed using Gas Chromatograph with high-resolution Mass Spectrometer (GC-HRMS) to identify the presence of active compounds. In vitro bioassays were performed using larval packet test (LPT) and adult immersion test (AIT) by taking different concentrations of CSO (i.e. 0.1, 0.5, 1.5, 2.5, 5, 7.5, 10 and 12.5%). In vivo acaricidal activity of CSO was evaluated by its topical application on red Sahiwal calves for 144 h. Clinical safety of CSO was evaluated by performing skin irritancy test and examination of hematological profile of calves'. GC-HRMS analysis of CSO revealed the presence of many fatty acids including oleic acid, lauric acid, palmitic acid, stearic acid and other components. Results exhibited that all the concentrations of CSO were effective in reducing the number of ticks and their growth. However, CSO at concentrations of 10% (CSO7) and 12.5% (CSO8) exhibited 100% mortality of R. microplus larvae and adults in LPT and AIT, respectively. In vivo acaricidal assay revealed that CSO7 and CSO8 shown 85% and 89% inhibition of ticks, respectively on calves after 144 h as compared to the control group. CSO was clinically safe on calves' skin with mild erythema up to 20 min. Hematological profile of calves revealed no sign of toxicity after treatment with CSO. Thus, CSO can be used as an alternative and safe drug therapy against R. microplus.

A participatory surveillance of marsh deer (Blastocerus dichotomus) morbidity and mortality in Argentina: first results.

BACKGROUND: In an era of unprecedented socio-ecological changes, managing wildlife health demands high-quality data collection and the engagement of local communities. Blastocerus dichotomus, the largest South American deer, is Vulnerable to extinction mainly due to habitat loss. Diseases have been recognised as a potential threat, and winter mortality has been historically described in marsh deer populations from Argentina. Field difficulties have, however, prevented in-depth studies of their health status. RESULTS: Between May 2014 and April 2017, we investigated marsh deer morbidity and mortality in the two largest populations in Argentina. We collected data by means of a passive surveillance system that involved a network of researchers, field partners (veterinarians, park rangers, and local community), and decision makers. We sampled marsh deer during as well as outside mortality events. A total of 44 marsh deer with different body condition scores were evaluated. We obtained haematology and biochemistry values from animals with good body condition score. Marsh deer with poor body condition had a high burden of the ticks Amblyomma triste and Rhipicephalus microplus. Vector-borne agents such as Theileria cervi, Trypanosoma theileri, Trypanosoma evansi, Ehrlichia chaffeensis, Anaplasma platys, Anaplasma odocoilei, Anaplasma marginale, and Candidatus Anaplasma boleense were also found. Haemonchus spp., Ostertagia spp., and Trichostrongylus spp. were the most frequent gastrointestinal parasites in deer with poor body condition. A Multiple Correspondence Analysis reinforced a possible association of winter period with lower body score condition, high tick loads, infection with E. chaffeensis, and presence of harmful gastrointestinal parasites. CONCLUSIONS: Our approach allowed the establishment of a participatory surveillance network of marsh deer morbidity and mortality in Argentina. We report and analyse the first data obtained opportunistically within the framework of this network, providing information on the infectious and parasitic agents in marsh deer populations. The occurrence of Fasciola hepatica and Leptospira interrogans serovar pyrogenes is reported for the first time in wild marsh deer from Argentina. Our data will be useful to improve the interpretation of future mortality events. The field implementation of a surveillance network is key to a holistic approach to wildlife diseases.

The potential of Allium sativum and Cannabis sativa extracts for anti-tick activities against Rhipicephalus (Boophilus) microplus.

The efficacy of Allium sativum and Cannabis sativa against Rhipicephalus microplus ticks was evaluated using the adult immersion and the larval packet test. In addition, an in silico approach was utilized by performing a docking study in order to identify the active ingredients from both plants. Results showed a comparatively high lethal effect of A. sativum and C. sativa on egg laying (index of egg laying = 0.26 and 0.24, respectively), egg hatching (33.5 and 37.1, respectively), and total larval mortality (100%, both), at 40 mg/mL. When applied to cattle which had been inoculated with larvae ticks, it was observed that a 45% solution of both herbal extracts significantly reduced the number of ticks by 96 h post treatment. We analyzed in silico 27 known active molecules from both plants and identified in the PubChem database to explore the hypothesis that the effect found on ticks was based on inhibition of acetylcholinesterase (AChE). Vitamin E and cannabidiol are the most potent AChE inhibitors with docking scores of -15.85 and -14.38, respectively. Based on these findings, we conclude that A. sativum and C. sativa may potentially be used for the control of R. microplus, and should be further investigated as a potential supplement to or replacement of synthetic acaricides.

In vitro acaricidal activity of Cymbopogon citratus, Cymbopogon nardus and Mentha arvensis against Rhipicephalus microplus (Acari: Ixodidae).

The chemical composition and acaricidal activity of plant-derived essential oils was assessed against Rhipicephalus microplus ticks. The essential oils of Mentha arvensis, Cymbopogon citratus and C. nardus were assessed for acaricidal activity against Rhipicephalus microplus. Essential oils (EO) of plants were separated by hydrodistillation (three times) and analyzed using gas chromatography - mass spectrometer (GC-MS). For bioassays, engorged females of R. microplus were exposed to C. citratus and C. nardus EO at 2%, 3%, 4% and 5% concentrations; and to M. arvensis EO at 1%, 3%, and 5% for 5 min. The weight egg mass, nutrient index (N.I), egg production index (E.P.I), hatching and control rate were evaluated. Non-feed larvae of R. microplus were exposed to essential oils with 0.25%, 0.5%; 1%; 1.5% and 2% concentrations; the mortality rate was measured after 48 h. Only engorged females presented reduced biological activities (oviposition, E.P.I) after exposure to M. arvensis at 3%, when in comparison to both positive and negative controls. The hatchability of R. microplus larvae ranged from 66.9% (after exposure to C. nardus EO at 5%) to 99.2% (positive control). The nutrition index was lower (46.6%) for the exposure to M. arvensis EO at 5%. M. arvensis at 3% and 5% concentrations was significantly efficient for engorged females when compared to control (53.7% and 47.5%, respectively). C. citratus EO at 1%, 1.5% and 2% concentrations yielded better results in the larval packet test, causing 100% mortality. Nonetheless, C. nardus and M. arvensis EO at 2% yielded 66% and 39% mortality, respectively. The study showed that M. arvensis presented potential for the control of R. microplus engorged females while C. citratus and C. nardus presented potential as a larvicide.

Repellency effect of Pilocarpus spicatus A. St.-Hil essential oil and nanoemulsion against Rhipicephalus microplus larvae.

Rhipicephalus microplus, the cattle tick, is a major cause of economic losses in bovine production. Due to the widespread acaricidal resistance to commercially available products, as well as their toxicity and environmental impact, alternative control methods are required. Nanoformulations produced from plant extracts as bioactive substances are very promising as innovative acaricidal agents. Thus, the aim of this study was to evaluate the in vitro repellent activity of Pilocarpus spicatus essential oil and its nanoemulsion against R. microplus, using larval repellent test (RT). The essential oil was extracted by hydrodistillation, using a Clevenger-type apparatus. The nanoemulsion was prepared with 5% essential oil, 5% tween 80, and 90% water, using the phase inversion method (50 mg/mL). Limonene was the major component (46.8%) of the essential oil, as determined by gas chromatography-mass spectrometry (GC/MS) and confirmed by flame ionization detection (GC/FID). According to the RT results, the essential oil had a repellent activity greater than 69%, from concentrations of 3.12 mg/mL (69.81 ± 10%) to 50 mg/mL (98.10 ± 0.6%), whereas the nanoemulsion at 50 mg/mL presented repellent activities of 97.14 ± 1.37% and 97.89 ± 0.52% 6 and 10 h after treatment, respectively. These values regarding to total repellency were very close to those calculated for mortality corrected by Abbott's formula. The phase inversion method preserved the chemical and physical characteristics of the essential oil since both reached an equal repellent effect at the same concentration. Therefore, P. spicatus essential oil and nanoemulsion had excellent repellent activities against R. microplus larvae, demonstrating its potential for future use as an alternative for tick control.

Botanical acaricides and repellents in tick control: current status and future directions.

Ticks are obligate blood-sucking ectoparasites and notorious as vectors of a great diversity of, in many instances, zoonotic pathogens which can cause considerable damage to animal and human health. The most commonly used approach for the control of ticks is the application of synthetic acaricides. However, the negative impacts of synthetic acaricides on the treated animals and the environment, in addition to its documented role in the development of resistance has led to the search for safer and more environmentally friendly alternative methods without compromising efficacy. An emerging promising approach for the control of ticks which has attracted much attention in recent years is the use of botanicals. Indeed, botanicals have been widely reported to show diverse effects and great potential as tick repellent and control. Although several excellent reviews have previously focused on this topic, studies on the exploration and application of botanicals to control ticks have expanded rapidly. Herein, we provide an update on the current understanding and status of botanical acaricides and repellents in tick control using recently published articles between 2017 and 2019. We also discuss the challenges and future directions in the application of botanicals in tick control, with a view of providing important clues for designing new integrated tick control methods.

A comparison of the efficacy of two commercial acaricides (fipronil and amitraz) with Azadirachta indica (neem) on the brown dog tick (Rhipicephalus sanguineus) from canines in Trinidad.

The brown dog tick (Rhipicephalus sanguineus) is prevalent on canids in Trinidad. It is directly (by causing anaemia) and indirectly (by acting as a vector of tick-borne pathogens) responsible for morbidity and mortalities in the canine population. The most commonly used commercial acaricides available to pet owners in Trinidad are amitraz and fipronil. Often, these acaricides may be abused and misused in a desperate attempt to rid pets of ticks. The objective of this study was to compare the efficacy of amitraz and fipronil with the herbal alternative, neem (Azadirachta indica). Triplicate in vitro trials utilizing the Larval Packet Test (LPT) were conducted using three concentrations (low, recommended and high) of fipronil (0.025%, 0.05% and 0.1%), amitraz (0.01%, 0.02% and 1%), neem oil (10%, 20% and 40%) and neem leaf extract (0.25%, 0.5% and 2%) for each trial. Statistical analysis using the mixed-effect Poisson regression analysis indicated that there was a significant difference (p < .05) in the survival of ticks pre-treatment versus post-treatment with amitraz, fipronil and all controls when compared to the neem oil. Fipronil and amitraz caused ≥99% mortality for all concentrations used in this study. Mortalities for neem oil and neem leaf extract ranged from 72.7% to 82% and 38% to 95.3%, respectively, with the greatest percentage of mortalities occurring at the lower concentrations. Neem oil and neem leaf extract can be used as alternative acaricides, and however, they are less efficacious against the brown dog tick than amitraz and fipronil.

Utilisation of indigenous knowledge to control ticks in goats: a case of KwaZulu-Natal Province, South Africa.

Local indigenous knowledge (IK) informs decision-making about fundamental aspects of life. The inclusion of indigenous knowledge is increasingly becoming a topical subject to enhance livestock veterinary care. The objective of the current study was to explore the extent of utilisation of indigenous knowledge to control ticks in goats. There was an association (P < 0.05) between the use of IK and gender, with males using IK (76.58%) more than females. The association between age distribution and IK use was (P < 0.05); however, farmers above the age of 50 years were using IK more than all group ages. Farmers ranked the purposes of using IK differently (P < 0.05). Ectoparasites were ranked as the most important constraint limiting goat productivity. Ticks were ranked as the most important external parasites. Amblyomma tick species were ranked as the most important amongst the tick species, followed by Rhipicephalus evertsi evertsi ticks. A significant population of farmers (80.7%) are dependent on the use of tick sprays, whereas others used injections (3.3%). Cissus quadrangularis L. (Inhlashwana) was singled out as the most used ethno-veterinary plant to control ticks with a frequency of (64%), followed by Gomphocarpus physocarpus E. Mey (Uphehlacwathi) (55.9%). The probability of keeping goats in wet rangelands (P < 0.05) was 3.04 times more likely to influence the extent of IK use compared to their contemporaries in the dry rangeland. Male farmers keeping goats (P < 0.01) were 2.95 more likely to influence the extent of use of IK than females. The type of rangeland, gender, age, residing on farm and also having the herbalist in the locality were the most common factors that influenced the extent of IK utilisation.

Development of essential oil-based phyto-formulations to control the cattle tick Rhipicephalus microplus using a mixture design approach.

The cattle tick Rhipicephalus (Boophilus) microplus is one of the most important ectoparasites for livestock in tropical and subtropical areas around the world. This tick economically impacts cattle production by reducing weight gain and milk production. Moreover, it is a vector of pathogens causing diseases such as babesiosis and anaplasmosis. Conventional tick control relies mainly on the use of chemical acaricides; however, their intensive use has led to the rapid appearance of resistant tick populations. It is therefore necessary to look for alternative tick control products. In that sense, plant extracts might represent a promising source of new acaricides. Previously, we reported a significant acaricide effect of essential oils from selected plant species. In the present study, we used a mixture design approach to develop phyto-formulations by combining individual essential oils. We produced several mixtures at 10% containing different proportions of individual essential oils (ranging from 0 to 1) from cinnamon (Cinnamomum zeylanicum), cumin (Cuminum cyminum) and allspice (Pimenta dioica) and tested their acaricidal activity against R. microplus ticks by means of larval packet test (LPT) and adult immersion test (AIT) assays. The optimal mixture predicted against R. microplus was composed of 66%, 17% and 17% of essential oils from C. zeylanicum, C. cyminum and P. dioica, respectively. We generated an estimated response surface contour plot that estimates 80%-100% acaricidal efficacy. In the optimal mixture 34 compounds were identified, which represent 98.65% of the total composition, with cinnamaldehyde (37.77%), ß-caryophyllene (13.92%), methyl eugenol (12.27%) and cuminaldehyde (8.99%) being the major components. Next, we developed emulsions by combining the optimal mixture with several surfactants and determined particle size, Zeta potential, stability and bioactivity. Emulsions containing 2% and 5% Tween 20 or Tween 80 remain stable after 14 days at 54 °C. Finally, optimized emulsion retained a high acaricidal activity against larval and adult R. microplus ticks. Taken together, our findings showed the usefulness of mixture design method for the development of essential oil mixtures with potent acaricidal activity. These formulations have the potential to successfully control tick infestations.

Sublethal concentrations of acetylcarvacrol strongly impact oocyte development of engorged female cattle ticks Rhipicephalus microplus (Canestrini, 1888) (Acari: Ixodidae).

Rhipicephalus microplus, commonly known as southern cattle tick, causes huge economic losses in the cattle industry. Its infestation affects the production of meat and milk and causes discomfort to hosts. In addition, it is the vector of Babesia spp. and Anaplasma spp. The most frequent method used to control these parasites is through synthetic acaricides. However, their indiscriminate use can be toxic for hosts and environment as well as cause selection of resistant ticks. Plant extracts and essential oils emerge as promising alternatives to manage tick infestation. Carvacrol, an aromatic monoterpene extracted from plants, has recognized antimicrobial, antioxidant, insecticidal, repellent and acaricidal activities. Acetylation of carvacrol is believed to enhance its nematicidal and acaricidal activities and to decrease its toxicity to hosts. Thus, the aim of this study was to evaluate the effect of different concentrations of acetylcarvacrol in the morphology of ovaries of engorged R. microplus ticks. The most remarkable morphological alterations found in the female germ cells were irregular and thicker chorion, decreasing in size and irregular shape of female germ cells (oocytes), cytoplasmatic vacuolization as well as ring-shaped nucleoli. These alterations were analyzed through a semi-quantitative method proposed in this study for ixodid ticks. Treatment group IV, which was exposed to 4.5 µL/mL of carvacrol acetate, showed the most significant alterations, and it was also statistically different when compared to control groups. Therefore, sublethal concentrations of acetylcarvacrol demonstrated to impact the reproductive system of R. microplus by causing several damages in the female germ cells. This would hinder the generation of new individuals, probably contributing for a long-term control of tick infestation.