Current perspectives and difficulties in the design of acaricides and repellents from plant-derived compounds for tick control.

Ticks and tick-borne diseases have gained increasing attention in recent years due to their impact on public health and significant losses in livestock production. The use of synthetic compounds for tick control is becoming problematic, mainly due to the resistance to commercially available products as well as their toxicity. Therefore, new alternative control methods are required. For this purpose, plant-derived extracts may be considered as effective repellents and/or acaricides. The present literature review focuses on studies evaluating the acaricidal and repellent activity of plant-derived extracts and plant secondary metabolites. We also noted recent advances in protein-ligand-docking simulation to examine the possible toxic effect of natural chemical compounds on ticks. In conclusion, plant-derived repellents/acaricides can be effective against ticks, especially in rural areas and livestock farms.

Novel ß-Cyclodextrin and Catnip Essential Oil Inclusion Complex and Its Tick Repellent Properties.

Cyclodextrin inclusion complexes have been successfully used to encapsulate essential oils, improving their physicochemical properties and pharmacological effects. Besides being well-known for its effects on cats and other felines, catnip (Nepeta cataria) essential oil demonstrates repellency against blood-feeding pests such as mosquitoes. This study evaluates the tick repellency of catnip oil alone and encapsulated in ß-cyclodextrin, prepared using the co-precipitation method at a 1:1 molar ratio. The physicochemical properties of this inclusion complex were characterized using GC-FID for encapsulation efficiency and yield and SPME/GC-MS for volatile emission. Qualitative assessment of complex formation was done by UV-Vis, FT-IR, 1H NMR, and SEM analyses. Catnip oil at 5% (v/v) demonstrated significant tick repellency over time, being comparable to DEET as used in commercial products. The prepared [catnip: ß-CD] inclusion complex exerted significant tick repellency at lower concentration of the essential oil (equivalent of 1% v/v). The inclusion complex showed that the release of the active ingredient was consistent after 6 h, which could improve the effective repellent duration. These results demonstrated the effective tick repellent activity of catnip essential oil and the successful synthesis of the inclusion complex, suggesting that ß-CDs are promising carriers to improve catnip oil properties and to expand its use in repellent formulations for tick management.

Acaricidal activity of leaves of Morus nigra against the cattle tick Rhipicephalus microplus / Atividade acaricida das folhas de Morus nigra em carrapato Rhipicephalus microplus

The acaricidal activity of crude ethanolic extract and fractions from the leaves of Morus nigra (Moraceae) was carried out on female cattle ticks Rhipicephalus microplus, using the adult immersion test. The mortality and fertility of females exposed to different concentrations of hexane, chloroform and ethyl acetate fractions, as well as ethanolic extract of M. nigra with concentrations of 5, 10 and 25mg/ml were evaluated using three treatment groups, two control groups and triplicate tests. The study also identified the main phenolic compounds of the extract and fractions of this species by HPLC. The chloroform fraction of leaves of M. nigra (25mg/mL) showed the best results for this species, obtaining 62.6% of inhibition of oviposition, 39.3% of eggs eclosion average and 65.4% of effectiveness. HPLC analysis revealed the presence of phenolic compounds, which may be related to biological activity shown by the extracts, which can be used as an alternative control against R. microplus adult tick.(AU)

A atividade acaricida do extrato etanólico bruto e das frações das folhas de Morus nigra (Moraceae) foi avaliada no carrapato Rhipicephalus microplus, utilizando-se o teste de imersão de fêmeas ingurgitadas. Foram avaliadas a mortalidade e a fertilidade das fêmeas ingurgitadas, expostas a diferentes concentrações (5, 10 e 25 mg/mL) das frações hexânica, clorofórmica e acetato de etila, bem como do extrato etanólico. Os testes foram realizados em triplicata. O estudo também identificou os principais compostos fenólicos do extrato e das frações dessa espécie por HPLC. A fração clorofórmica das folhas de M. nigra (25mg/mL) apresentou o melhor resultado para essa espécie, obtendo-se 62,6% de inibição da oviposição, 39,3% da eclosão dos ovos e 65,4% de eficácia. A análise por HPLC revelou a presença de compostos fenólicos, os quais podem estar relacionados com a atividade biológica demonstrada pelos extratos, que podem ser utilizados como auxiliar no controle contra carrapato adulto de R. microplus.(AU)

Plant products and secondary metabolites with acaricide activity against ticks.

The present review documents the results of studies evaluating the acaricidal activity of different plant products and secondary metabolites against ticks that are resistant and susceptible to conventional acaricides. Studies published from 1998 to 2016 were included. The acaricidal activity of plant extracts, essential oils and secondary compounds from plants have been evaluated using bioassays with ticks in the larval and adult stages. There is variable effectiveness according to the species of plant and the concentrations used, with observed mortalities ranging from 5 to 100% against the Rhipicephalus (Boophilus), Amblyomma, Dermacentor, Hyalomma, and Argas genera. A number of plants have been reported to cause high mortalities and/or affect the reproductive capacity of ticks in the adult phase. In the majority of these trials, the main species of plants evaluated correspond to the families Lamiaceae, Fabaceae, Asteraceae, Piperaceae, Verbenaceae, and Poaceae. Different secondary metabolites such as thymol, carvacrol, 1,8-cineol and n-hexanal, have been found to be primarily responsible for the acaricidal activity of different essential oils against different species of ticks, while nicotine, dibenzyldisulfide and dibenzyltrisulfide have been evaluated for plant extracts. Only thymol, carvacrol and 1,8-cineol have been evaluated for acaricidal activity under in vivo conditions. The information in the present review allows the conclusion that the secondary metabolites contained in plant products could be used as an alternative for the control of ticks that are susceptible or resistant to commercial acaricides.

In vitro assessment of the acaricidal activity of Piper longum, Piper nigrum, and Zingiber officinale extracts against Hyalomma anatolicum ticks.

Ticks and tick-borne diseases are a major constraint for the sustainable cattle industry in the tropical and subtropical regions including the Indian subcontinent. The development of resistance to most of the commonly used acaricides leads to an attempt to screen plant extracts and their combinations for their possible acaricidal activity to develop an eco-friendly tick control alternative. An alcoholic and various aqueous extracts of Piper longum, Piper nigrum and Zingiber officinale and their combinations were evaluated for acaricidal activity against the three-host ixodid tick, Hyalomma anatolicum by larval immersion test using 14-21 days old unfed larvae. The efficacy was assessed by measuring larval mortality (%) and the lethal concentrations for 50% (LC50) and 95% (LC95) with their 95% confidence limits (CL) values were estimated by applying regression equation analysis to the probit transformed data of mortality. A concentration-dependent mortality response was recorded in all extracts prepared from seeds of P. longum and P. nigrum and their combinations. The highest acaricidal property was exhibited by the alcoholic extract of P. longum seeds with the minimum LC50 and LC95 (95% CL) values of 0.071% (0.07-0.072) and 0.135% (0.13-0.14), respectively, followed by alcoholic combinations. Interestingly, no acaricidal activity was recorded in extracts prepared from the rhizome of Z. officinale. The results indicated that the ethanolic extracts of P. longum and P. nigrum and their combinations can be used effectively for tick control in an integrated format.

Application of ethnobotanical repellents and acaricides in prevention, control and management of livestock ticks: A review.

Ticks transmit at least the same number or even more pathogens than any other group of blood-feeding arthropods worldwide affecting humans and animals. The eco-friendly control and management of tick vectors in a constantly changing environment is a crucial challenge. Besides the development of vaccines against ticks, IPM practices aimed at reducing tick interactions with livestock, emerging pheromone-based control tools, and few biological control agents, the extensive employment of acaricides and tick repellents still remain the most effective and ready-to-use strategies. However, the former is limited by the development of growing resistances as well as environmental concerns. Exploiting plants and plant products as sources of effective tick repellents and acaricides represents a promising strategy. In this scenario, the preservation of ethnobotanical information on repellent and acaricidal potential of plants is crucial. Here, we evaluated relevant information published in recent years, focused on plants used as repellents and acaricides against tick vectors in different regions worldwide. We selected a total of 238 plant species, which are traditionally used against ticks by native and local communities of Africa (Kenya, Uganda, Zimbabwe, South Africa), Europe (Serbia, Macedonia, Romania), Asia (Pakistan, India) and America (Brazil, Canada), from 56 families. However, only 7 families (i.e. Asteraceae, Euphorbiaceae, Fabaceae, Lamiaceae, Meliaceae, Apocynaceae and Solanaceae) represent the major quote (46%) of all plant species. We evaluated the differences in acaricidal and repellent efficacy of different formulations used. In the final section, implications arising from the surveyed anti-tick ethnobotanical knowledge and challenges for its future are discussed.

Acaricidal activity of ethanolic extract of Artemisia absinthium against Hyalomma anatolicum ticks.

The aim of the study was to evaluate the in vitro efficacy of different concentrations of ethanolic extract obtained from the aerial parts of Artemisia absinthium in comparison to amitraz on adults, eggs and larvae of Hyalomma anatolicum using the adult immersion test (AIT), egg hatchability test and larval packet test (LPT), respectively. Four concentrations of the extract (2.5, 5, 10 and 20%) with three replications for each concentration were used in all the bioassays. In AIT, the mortality rates at 2.5, 5 and 10% were significantly different (p < 0.05) in comparison to the control group; however, at 20%, it was similar to the positive control group. Maximum mortality of 86.7% was recorded at 20%. The LC50 and LC95 values were calculated as 6.51 and 55.43%, respectively. The oviposition was reduced significantly by 36.8 and 59.1% at concentrations of 10 and 20%, respectively. Egg hatchability was reduced significantly at all concentrations (2.5-20%) in comparison to the control. In LPT, the extract caused 100% mortality of larvae at all the concentrations after 24 h. The results show that ethanolic extract obtained from the aerial parts of A. absinthium has acaricidal properties and could be useful in controlling H. anatolicum.

Tick repellents for human use: prevention of tick bites and tick-borne diseases.

Ticks are arthropods and the most important vectors of major human diseases after mosquitoes. Due to their impact on public health, in vitro and in vivo assays have been developed to identify molecules with repellent activities on ticks. Repellents are useful to reduce tick bite exposure and the potential transmission of pathogens; they can be used topically or in impregnated clothing. Presently, mainly synthetic molecules are commercialized as skin repellents, e.g., N,N-diethyl-meta-toluamide (DEET), IR3535, picaridin or KBR 3023, and para-menthanediol. Permethrin is largely used for fabric impregnation. Intensive research has been conducted to identify new molecules with repellent activity and more recently, plant-derived molecules, as an alternative to synthetic molecules.

Morphological alterations of epidermis of rabbits infested by R. sanguineus ticks and exposed to Selamectin (active principle of Pfizer Revolution(®) acaricide): a confocal microscopy study.

The present study analyzed, by means of confocal laser scanning microscopy, the epidermis of rabbits infested by the brown dog tick Rhipicephalus sanguineus and exposed to concentrations of 50%, 80% and 100% of a selamectin-based commercial acaricide (Pfizer's Revolution(®)). The results demonstrated that rabbits exposed to concentrations of 80% and 100% of the Revolution acaricide, which contains 12% selamectin, showed thinning of epithelial tissue of the epidermis with associated cellular disorganization. Individuals exposed to a 50% concentration showed lower epidermal tissue disorganization when compared to those exposed to the higher doses of the acaricide (80% and 100%). Whereas selamectin, when used in higher concentrations (80% and 100% Revolution(®)) can alter the morphology of the epidermis, at lower concentrations (50%), even though still able to eliminate ectoparasites, it causes less toxicity damage to the host. Selamectin can be considered a dose-dependent toxic agent, since higher concentrations increase the morphological changes in the epidermis of the host rabbits.

Bioactivity-guided investigation of geranium essential oils as natural tick repellents.

The evaluation of 10 essential oils of geranium, Pelargonium graveolens (Geraniaceae), were all shown to have repellent activity against nymphs of the medically important lone star tick, Amblyomma americanum (L.). The biological tests were carried out using a vertical filter paper bioassay, where ticks must cross an area of the paper treated with repellent to approach host stimuli. One of the essential oil samples that repelled >90% of the ticks at 0.103 mg/cm(2) was selected for further fractionation studies. The sesquiterpene alcohol, (-)-10-epi-γ-eudesmol, was isolated and identified by spectral methods. (-)-10-epi-γ-Eudesmol at 0.103 and 0.052 mg of compound/cm(2) of filter paper repelled 90 and 73.3% of the ticks, respectively. (-)-10-epi-γ-Eudesmol exhibited similar repellency to the reference standard N,N-diethyl-meta-toluamide (DEET) at concentrations of ≥0.052 mg of compound/cm(2) of filter paper, with (-)-10-epi-γ-eudesmol losing much of its repellency at 0.026 mg of compound/cm(2) and DEET at 0.013 mg of compound/cm(2). Isomenthone and linalool did not repel ticks at the concentrations tested. Most repellents are marketed with much higher concentrations of active ingredient than the concentrations of the natural repellents tested herein; therefore, effective compounds, such as (-)-10-epi-γ-eudesmol, found in geranium oil, have the potential for commercial development.

Acetylcholinesterases of blood-feeding flies and ticks.

Acetylcholinesterase (AChE) is the biochemical target of organophosphate (OP) and carbamate pesticides for invertebrates, vertebrate nerve agents, and AChE inhibitors used to reduce effects of Alzheimer's disease. Organophosphate pesticides (OPs) are widely used to control blood-feeding arthropods, including biting flies and ticks. However, resistance to OPs in pests affecting animal and human health has compromised control efficacy. OP resistance often results from mutations producing an OP-insensitive AChE. Our studies have demonstrated production of OP-insensitive AChEs in biting flies and ticks. Complementary DNA (cDNA) sequences encoding AChEs were obtained for the horn fly, stable fly, sand fly, and the southern cattle tick. The availability of cDNA sequences enables the identification of mutations, expression and characterization of recombinant proteins, gene silencing for functional studies, as well as in vitro screening of novel inhibitors. The southern cattle tick expresses at least three different genes encoding AChE in their synganglion, i.e. brain. Gene amplification for each of the three known cattle tick AChE genes and expression of multiple alleles for each gene may reduce fitness cost associated with OP-resistance. AChE hydrolyzes the neurotransmitter, acetylcholine, but may have additional roles in physiology and development. The three cattle tick AChEs possess significantly different biochemical properties, and are expressed in neural and non-neural tissues, which suggest separation of structure and function. The remarkable complexity of AChEs in ticks suggested by combining genomic data from Ixodes scapularis with our genetic and biochemical data from Rhipicephalus microplus is suggestive of previously unknown gene duplication and diversification. Comparative studies between invertebrate and vertebrate AChEs could enhance our understanding of structure-activity relationships. Research with ticks as a model system offers the opportunity to elucidate structure-activity relationships for AChE that are important for advances in targeted pest control, as well as potential applications for medicine and biosecurity.

Natural product studies of U.S. endangered plants: volatile components of Lindera melissifolia (Lauraceae) repel mosquitoes and ticks.

The number of endangered plant species in the U.S. is significant, yet studies aimed towards utilizing these plants are limited. Ticks and mosquitoes are vectors of significant pathogenic diseases of humans. Repellents are critical means of personal protection against biting arthropods and disease transmission. The essential oil and solvent extracts from Lindera melissifolia (Walt.) Blume (Lauraceae) (pondberry) drupes were gathered and analyzed by GC and GC-MS. The essential oil obtained from this endangered plant showed a significant dose dependent repellency of ticks and a moderate mosquito repellent effect while the subsequent hexanes extract was completely ineffective. Fractional freezing enriched the tick repellent components of the essential oil. Several known tick repellent components were recognized by the GC-MS comparison of the resulting fractions and ß-caryophyllene, α-humulene, germacrene D and ß-elemene warrant evaluations for tick repellency. Identifying pondberry as a potential renewable source for a broad spectrum repellent supports efforts to conserve similar U.S. endangered or threatened plant species.

Evaluation of different extracts and synthesised silver nanoparticles from leaves of Euphorbia prostrata against Haemaphysalis bispinosa and Hippobosca maculata.

The present study was based on assessments of the antiparasitic activity to determine the efficacies of hexane, chloroform, ethyl acetate, acetone, methanol and aqueous leaf extracts of Euphorbia prostrata Ait. (Euphorbiaceae) and synthesised Ag nanoparticles (NPs) using aqueous leaf extract against the adult cattle tick Haemaphysalis bispinosa Neumann (Acarina: Ixodidae) and the haematophagous fly Hippobosca maculata Leach (Diptera: Hippoboscidae). Synthesised Ag NPs were characterised with ultraviolet-vis (UV-vis) spectrum, scanning electron microscopy (SEM), X-ray diffraction (XRD) and Fourier transform infrared (FTIR) support the biosynthesis of Ag NPs. Parasites were exposed to varying concentrations of plant extracts and synthesised silver NPs for 24 h. All extracts showed the maximum toxic effect on parasites; however, the highest mortality was found in the hexane, chloroform, ethyl acetate, acetone, methanol and aqueous leaf extracts of E. prostrata and synthesised Ag NPs against the adult of H. bispinosa (LC(50)=45.24, 40.07, 21.91, 25.32, 19.30, 10.16 and 2.30 ppm; LC(90)=86.95, 88.66, 70.92, 83.22, 48.28, 70.27 and 8.28 ppm) and against H. maculata (LC(50)=39.37, 41.98, 19.92, 27. 93, 21.97, 9.79 and 2.55 ppm; LC(90)=89.44, 98.52, 76.59, 90.18, 55.07, 54.35 and 9.03 ppm), respectively. Mortality of 100% was found in synthesised Ag NPs at a concentration of 10 mg l(-1). UV-vis spectrograph of the colloidal solution of Ag NPs has been recorded as a function of time. The absorption spectrum of E. prostrata leaf extracts at different wavelengths ranging from 300 to 600 nm revealed a peak at 420 nm after 6 h. The FTIR spectra of Ag NPs exhibited prominent peaks at 3431; 1616; 1381; 1045; 818; 509; and 420 cm(-1). SEM analyses of the synthesised Ag NPs were rod shaped and measured 25-80 nm with an average size of 52.4 nm. The chemical composition of aqueous leaf extract was analysed by gas chromatography-mass spectrometry (GC-MS). The major chemical constituent was identified as 2-phenylethanol. These results suggest that the leaf methanol, aqueous extracts of E. prostrata and green synthesis of Ag NPs have the potential to be used as an ideal eco-friendly approach for the control of H. bispinosa and H. maculata. In addition, toxicity tests were conducted to analyse the toxicological effects of particle size on Daphnia magna and Ceriodaphnia dubia, and the animal model test was evaluated against Bos indicus for 24-h treatment. No toxicity on daphnids and no adverse effects were noted on animals after exposure to solvent extracts and synthesised Ag NPs.

Evaluation of medicinal plant extracts against ticks and fluke.

The present study was based on assessments of the antiparasitic activities to determine the efficacies of leaf hexane, chloroform, ethyl acetate, acetone and methanol extracts of Aegle marmelos (Linn.) Correa ex Roxb, Andrographis lineata Wallich ex Nees., Andrographis paniculata (Burm.f.) Wallich ex Nees., Cocculus hirsutus (L.) Diels, Eclipta prostrata L., and Tagetes erecta L. against the adult cattle tick Haemaphysalis bispinosa Neumann 1897 (Acarina: Ixodidae), the larvae of Rhipicephalus (Boophilus) microplus Canestrini 1887 (Acari: Ixodidae) and sheep fluke Paramphistomum cervi Zeder 1790 (Digenea: Paramphistomatidae). All plant extracts showed moderate toxic effect on parasites after 24 h of exposure; however, the highest parasitic activity was found in leaf ethyl acetate extract of A. lineata, methanol extract of A. marmelos, A. paniculata, and C. hirsutus against H. bispinosa (LC(50) = 395.27, 358.45, 327.21 and 420.50 ppm); ethyl acetate extract of A. paniculata, C. hirsutus, methanol extracts of A. marmelos, A. lineata, and E. prostrata against the larvae of R. microplus (LC(50) = 207.70, 258.61, 134.09, 206.00, and 274.33 ppm); hexane extract of A. lineata, ethyl acetate extract of A. paniculata, E. prostrata, acetone extracts of T. erecta, methanol extracts of A. marmelos and C. hirsutus against P. cervi (LC(50) = 254.23, 451.17, 425.73, 253.60, 542.71, and 360.17 ppm), respectively. The present study is the first report on the veterinary parasitic activity of plant extracts from Southern India.

Repellent activity of catmint, Nepeta cataria, and iridoid nepetalactone isomers against Afro-tropical mosquitoes, ixodid ticks and red poultry mites.

The repellent activity of the essential oil of the catmint plant, Nepeta cataria (Lamiaceae), and the main iridoid compounds (4aS,7S,7aR) and (4aS,7S,7aS)-nepetalactone, was assessed against (i) major Afro-tropical pathogen vector mosquitoes, i.e. the malaria mosquito, Anopheles gambiae s.s. and the Southern house mosquito, Culex quinquefasciatus, using a World Health Organisation (WHO)-approved topical application bioassay (ii) the brown ear tick, Rhipicephalus appendiculatus, using a climbing repellency assay, and (iii) the red poultry mite, Dermanyssus gallinae, using field trapping experiments. Gas chromatography (GC) and coupled GC-mass spectrometry (GC-MS) analysis of two N. cataria chemotypes (A and B) used in the repellency assays showed that (4aS,7S,7aR) and (4aS,7S,7aS)-nepetalactone were present in different proportions, with one of the oils (from chemotype A) being dominated by the (4aS,7S,7aR) isomer (91.95% by GC), and the other oil (from chemotype B) containing the two (4aS,7S,7aR) and (4aS,7S,7aS) isomers in 16.98% and 69.83% (by GC), respectively. The sesquiterpene hydrocarbon (E)-(1R,9S)-caryophyllene was identified as the only other major component in the oils (8.05% and 13.19% by GC, respectively). Using the topical application bioassay, the oils showed high repellent activity (chemotype A RD(50)=0.081 mg cm(-2) and chemotype B RD(50)=0.091 mg cm(-2)) for An. gambiae comparable with the synthetic repellent DEET (RD(50)=0.12 mg cm(-2)), whilst for Cx. quinquefasciatus, lower repellent activity was recorded (chemotype A RD(50)=0.34 mg cm(-2) and chemotype B RD(50)=0.074 mg cm(-2)). Further repellency testing against An. gambiae using the purified (4aS,7S,7aR) and (4aS,7S,7aS)-nepetalactone isomers revealed overall lower repellent activity, compared to the chemotype A and B oils. Testing of binary mixtures of the (4aS,7S,7aR) and (4aS,7S,7aS) isomers across a range of ratios, but all at the same overall dose (0.1 mg), revealed not only a synergistic effect between the two, but also a surprising ratio-dependent effect, with lower activity for the pure isomers and equivalent or near-equivalent mixtures, but higher activity for non-equivalent ratios. Furthermore, a binary mixture of (4aS,7S,7aR) and (4aS,7S,7aS) isomers, in a ratio equivalent to that found in chemotype B oil, was less repellent than the oil itself, when tested at two doses equivalent to 0.1 and 0.01 mg chemotype B oil. The three-component blend including (E)-(1R,9S)-caryophyllene at the level found in chemotype B oil had the same activity as chemotype B oil. In a tick climbing repellency assay using R. appendiculatus, the oils showed high repellent activity comparable with data for other repellent essential oils (chemotype A RD(50)=0.005 mg and chemotype B RD(50)=0.0012 mg). In field trapping assays with D. gallinae, addition of the chemotype A and B oils, and a combination of the two, to traps pre-conditioned with D. gallinae, all resulted in a significant reduction of D. gallinae trap capture. In summary, these data suggest that although the nepetalactone isomers have the potential to be used in human and livestock protection against major pathogen vectors, intact, i.e. unfractionated, Nepeta spp. oils offer potentially greater protection, due to the presence of both nepetalactone isomers and other components such as (E)-(1R,9S)-caryophyllene.

Potential of medicinal plants in mosquito control.

Medicinal plants have long history as important components in traditional medicine, and food of humans since ancient Egyptians and Chinese. Naturally occurring botanical compounds contain a broad range of chemical active ingredients can intervene in all biological processes of the mosquito, thus interrupt its life cycle and dispersal and reduce harms to humans and animals. Many medicinal plants were tested for their pesticide and repellent potential, as crude material, essential oils or individual active ingredients. This article reviewed studies on the efficacy of many well known and commonly used safe medicinal plants or their products in controlling the mosquitoes; Aedes aegypti, Anopheles gambiae, An. stephensi and Culex quinquefasciatus and the ticks, Dermacentor variabilis, Amblyomma americanum, Ixodes scapularis and I. ricinus. Promising and encouraging results were obtained against these arthropod-vectors of zoonotic diseases.

Extracts of Calpurnia aurea leaves from southern Ethiopia attract and immobilise or kill ticks.

Calpurnia aurea extracts are used in southern Ethiopia to protect stock against ticks. Acetone, hexane and water leaf extracts of C. aurea collected in southern Ethiopia were tested for repellent/attractant and acaricidal properties on unfed adult Rhipicephalus pulchellus ticks. In contrast to many other plant species evaluated, C. aurea extracts did not have repellent properties, but rather had a slight attractant capacity. With 20% and 10% acetone extracts, all ticks were either killed or their mobility severely compromised after 1microl of extract was topically applied on the abdomen. At a 5% concentration, 85% of ticks were still affected. A 10% aqueous solution also had a marked effect. The results prove the efficacy of the traditional use of this extract and may lead to a product that can be used commercially to protect animals against tick infestation, under subsistence as well as industrialized conditions.

Adulticidal and larvicidal efficacy of some medicinal plant extracts against tick, fluke and mosquitoes.

The adulticidal and larvicidal effect of indigenous plant extracts were investigated against the adult cattle tick Haemaphysalis bispinosa Neumann, 1897 (Acarina: Ixodidae), sheep fluke Paramphistomum cervi Zeder, 1790 (Digenea: Paramphistomatidae), fourth instar larvae of malaria vector, Anopheles subpictus Grassi and Japanese encephalitis vector, Culex tritaeniorhynchus Giles (Diptera: Culicidae). The aim of this study was to evaluate the toxic effect of leaf hexane, chloroform, ethyl acetate, acetone and methanol extracts of Annona squamosa L., Centella asiatica (L.) Urban, Gloriosa superba L., Mukia maderaspatensis (L.) M.Roem, Pergularia daemia (Forsk.) Chiov. and Phyllanthus emblica L. were exposed to different concentrations. All plant extracts showed moderate toxic effect on parasites after 24h of exposure; however, the highest mortality was found in leaf hexane extract of A. squamosa, methanol extracts of G. superba and P. emblica against H. bispinosa (LC(50)=145.39, 225.57 and 256.08ppm); methanol extracts of C. asiatica, G. superba, P. daemia and P. emblica against P. cervi (LC(50)=77.61, 60.16, 59.61, and 60.60ppm); acetone, ethyl acetate extracts of A. squamosa, methanol extract of C. asiatica, acetone extracts of G. superba, ethyl acetate, hexane and methanol extracts of P. daemia against A. subpictus (LC(50)=17.48, 18.60, 26.62, 18.43, 34.06, 13.63, and 50.39ppm); and chloroform, ethyl acetate extracts of A. squamosa, ethyl acetate extract of P. daemia, ethyl acetate and methanol extracts of P. emblica against C. tritaeniorhynchus (LC(50)=63.81, 60.01, 31.94, 69.09, and 54.82ppm), respectively. These results demonstrate that methanol extracts of C. asiatica, G. superba, P. daemia and P. emblica extracts may serve as parasites control even in their crude form.

Tick repellent substances in the essential oil of Tanacetum vulgare.

The repellent effect of the essential oils of flower heads of the aromatic plant tansy, Tanacetum vulgare L. (Asteraceae), originating from Sweden, was tested against host-seeking nymphs of the common tick Ixodes ricinus (L.). The essential oils were obtained by steam distillation (SD) and by using an online solvent extraction separation setup. Further fractionations of the SD oils were obtained by medium-pressure liquid chromatography on silica gel. The volatiles of the essential oils and the fractions that exhibited strong tick repellency (90-100%) were collected by solid phase microextraction and identified by gas chromatography-mass spectrometry. The chemical analyses of the oils show that the populations of T. vulgare from Uppsala and Stockholm may represent different chemotypes, but that they exhibited similar tick repellency. Main volatiles detected from oils of T. vulgare collected at Uppsala were alpha-pinene (27%), beta-pinene (11%), pinocamphone (11%), 1,3,3-trimethylcyclohex-1-ene-4-carboxaldehyde (11%), and 1,8-cineole (10%). In the sample collected in Stockholm, the main components were beta-thujone (39%) and camphor (23%) followed by alpha-thujone (11%) and 1,8-cineole (8%). When constituents in the oils, e.g., alpha-terpineol, 4-terpineol, alpha+beta-thujone, 1,8-cineol, verbenol, and verbenone, were tested separately (each diluted 0.5%, vol:vol), 64-72% tick repellency was obtained.

In vitro investigation of the toxic effects of extracts of Allium sativum bulbs on adults of Hyalomma marginatum rufipes and Rhipicephalus pulchellus.

The toxic effects of the extracts of Allium sativum (Garlic) were evaluated against adults of Hyalomma marginatum rufipes and Rhipicephalus pulchellus using three types (Types A, B and C) of contact toxicity bioassays. A. sativum bulbs were extracted with acetone, ethanol and dichloromethane (DCM) solvents. Among these three solvents, it is the DCM extract of A. sativum that appears to have anti-tick activity. In the Type A contact toxicity bioassay, DCM extracts of A. sativum demonstrated a high acaricidal bioactivity against H. m. rufipes with 100% of ticks killed in less than an hour, and toxicity persisted to the second day. A weak acaricidal activity of aqueous extracts of A. sativum was observed in the Type B contact toxicity bioassay. In the Type C contact toxicity bioassay, a concentration of 24% w/v of DCM extracts of garlic in sunflower oil (Helianthus annuus) had killed 100% of H. m. rufipes (LC50 = 5.9% w/v) and R. pulchellus (LC50 = 10.3% w/v) by 24 hours post-treatment of ticks. The results obtained from this study suggest that DCM extract of A. sativum is a potential source of novel acaricidal agents.