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Shabab Ahmad
Muhammad Oneeb
Muhammad Lateef
Muhammad Ijaz
Muhammad Irfan Siddique
Sajida Nawaz


Rhipicephalus microplus, Mentha piperita, Mentha piperitax.  acaricidal activity, Toxicity evaluation


Ticks and tick-borne diseases (TTBDs) are among the global health challenges. The irrational use of synthetic chemicals poses serious threats in terms of toxicity, environmental hazards and resistance. The use of essential oils as an augmentative approach can be highly important for combatting this issue. However, their volatile nature, low stability and direct exposure to extreme environmental conditions compromise their efficacy. The use of suitable polymers to encapsulate essential oils is one way forward. Mentha (M.) piperita essential oil (EO) is a possible alternative for the control of TTBDs. The aim of this study was to evaluate the in vitro acaricidal efficacy of chitosan (CS)-encapsulated M. piperita EO against R. microplus. The M. piperita EO was encapsulated in Preparation of CS nanoparticles (NPs) following emulsification/ionic gelation. The adult immersion bioassay was employed to analyse the acaricidal activity of encapsulated M. piperita EO (8 mg/ml to 0.5 mg/ml). Furthermore, the toxicity of M. piperita EO NPs against non-target species was evaluated by three tests including:  Acute dermal irritation test, acute dermal toxicity study and skin sensitization test. It exhibited excellent acaricidal activity against R. microplus, the efficacy of encapsulated M. piperita EO was the highest, increasing up to 100% (RC50=0.877, RC90=5.231) at the highest concentration. Similarly, the inhibition of oviposition was maximum at highest concentration. While, hatching rate of laid eggs increased with the decrease of concentration. Moreover, the M. piperita EO NPs were found nontoxic for non-target species. The present study provides the first description of the acaricidal activity of nanoencapsulated M. piperita EO against adult R. microplus, which is an eco-friendly alternate control strategy for ticks.

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1. Nepveu-Traversy M-E, Fausther-Bovendo H, Babuadze GJV. Human Tick-Borne Diseases and Advances in Anti-Tick Vaccine Approaches: A Comprehensive Review. 2024;12(2):141.
2. Reis AC, Konig IFM, Mendonça GHA, Lunguinho AdS, Cardoso MdG, Remedio RNJE, et al. Toxic effect of Callistemon viminalis essential oil on the ovary of engorged Rhipicephalus microplus female ticks (Acari: Ixodidae). 2023;90(1):99-117.
3. Cruz RR, García DID, Silva SL, Domínguez FR. Integrated management of the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) and the Acaricide resistance mitigation. Insecticides-Impact and Benefits of Its Use for Humanity: IntechOpen; 2021.
4. Jain P, Satapathy T, Pandey RKJB, biotechnology a. Rhipicephalus microplus: A parasite threatening cattle health and consequences of herbal acaricides for upliftment of livelihood of cattle rearing communities in Chhattisgarh. 2020;26:101611.
5. Jamil A, Yu Z, Wang Y, Xin Q, Gao S, Abdul Wahab M, et al. Tick-borne Rickettsia, Anaplasma, Theileria, and enzootic nasal tumor virus in ruminant, PET, and poultry animals in Pakistan. 2024;15:1359492.
6. Kebede A, Mitiku A, Tilahun A, Oli Y, Gutata MJW. Prevalence of Ixodidae Ticks and its Associated Risk Factors in Local Breed (Zebu) Cattle at Mao Komo Special District Benishangul Gumuz Regional States, Western Ethiopia. 2024;12:1-7.
7. Tagel W. Prevalence and Associated Risk Factors of Ixodid Ticks Infestation And Tick Borne Haemoparasites of Small Ruminants In Enarje Enawuga District, North Western Ethiopia 2022.
8. Reshma KR, Prakasan KJE, Letters AS. Synthetic Acaricides As A Promising Tool in Tick Control Program-The Present Scenario. 2020;7(2-2020):58-69.
9. Stafford KA, Coles GCCJADRMoDR, Volume 1. Drug resistance in ectoparasites of medical and veterinary importance. 2017:735-44.
10. Gonçalves IL, das Neves GM, Kagami LP, Eifler-Lima VL, Merlo AAJB, Chemistry M. Discovery, development, chemical diversity and design of isoxazoline-based insecticides. 2021;30:115934.
11. Rodriguez-Vivas RI, Jonsson NN, Bhushan CJPr. Strategies for the control of Rhipicephalus microplus ticks in a world of conventional acaricide and macrocyclic lactone resistance. 2018;117:3-29.
12. Pelletier J, Rocheleau J-P, Aenishaenslin C, Dimitri Masson G, Lindsay LR, Ogden NH, et al. Fluralaner baits reduce the infestation of Peromyscus spp. mice (Rodentia: Cricetidae) by Ixodes scapularis (Acari: Ixodidae) larvae and nymphs in a natural environment. 2022;59(6):2080-9.
13. Ren Y, Dou W, Wang JJ, Yuan GJPMS. Lethal and sublethal effects of fluralaner on the citrus red mite, Panonychus citri (McGregor). 2024.
14. van Asselt E, Pikkemaat M, Jansen L, Hoek-van den Hil E. Prioritisation of chemical substances for national monitoring: Applied to antibiotics, antiparasitics, carbamates and NSAIDs in bovine, porcine and poultry products. RIKILT Wageningen University & Research; 2019.
15. Rodrigues L, Giglioti R, Morita Katiki L, Franceschini Sarria AL, José Veríssimo CJIJoA. Screening of volatile compounds of essential oils with acaricidal activity in Rhipicephalus microplus larvae using a new in vitro evaluation method. 2024:1-9.
16. Valcárcel F, Olmeda AS, González MG, Andrés MF, Navarro-Rocha J, González-Coloma AJFiA. Acaricidal and insect antifeedant effects of essential oils from selected aromatic plants and their main components. 2021;3:662802.
17. Štrbac F, Krnjajić S, Stojanović D, Ratajac R, Simin N, Orčić D, et al. In vitro and in vivo anthelmintic efficacy of peppermint (Mentha x piperita L.) essential oil against gastrointestinal nematodes of sheep. 2023;10.
18. Isman MBJPr. Commercial development of plant essential oils and their constituents as active ingredients in bioinsecticides. 2020;19:235-41.
19. Assadpour E, Can Karaça A, Fasamanesh M, Mahdavi SA, Shariat-Alavi M, Feng J, et al. Application of essential oils as natural biopesticides; recent advances. 2023:1-21.
21. Wu Z, Tan B, Liu Y, Dunn J, Martorell Guerola P, Tortajada M, et al. Chemical composition and antioxidant properties of essential oils from peppermint, native spearmint and scotch spearmint. 2019;24(15):2825.
22. Katekar VP, Rao AB, Sardeshpande VRJSC, Pharmacy. A hydrodistillation-based essential oils extraction: A quest for the most effective and cleaner technology. 2023;36:101270.
23. Karpiński TMJB. Essential oils of Lamiaceae family plants as antifungals. 2020;10(1):103.
24. Hudz N, Kobylinska L, Pokajewicz K, Horčinová Sedláčková V, Fedin R, Voloshyn M, et al. Mentha piperita: essential oil and extracts, their biological activities, and perspectives on the development of new medicinal and cosmetic products. 2023;28(21):7444.
25. Mahendran G, Rahman LUJPR. Ethnomedicinal, phytochemical and pharmacological updates on Peppermint (Mentha× piperita L.)—A review. 2020;34(9):2088-139.
26. Sartoratto A, Machado ALM, Delarmelina C, Figueira GM, Duarte MCT, Rehder VLGJBJoM. Composition and antimicrobial activity of essential oils from aromatic plants used in Brazil. 2004;35:275-80.
27. Al-Marby A. Nematicidal and antimicrobial evaluation of extracts, nanosized materials and fractions of selected plants and the identification of the bioactive phytochemicals. 2017.
28. Rodrigues MMG. Control of the Root-Knot Nematode, Meloidogyne Incognita, Using Plant Bioproducts: Universidade do Minho (Portugal); 2022.
29. Pereira BDF. Activity of essential oils and hydroalcoholic extracts from 12 plants against food spoilage yeasts: evaluation of their potential as new biopesticides: Universidade do Minho (Portugal); 2021.
30. Raveau R, Fontaine J, Lounès-Hadj Sahraoui AJF. Essential oils as potential alternative biocontrol products against plant pathogens and weeds: A review. 2020;9(3):365.
31. Noaman V, Bahreininejad BJJoA-PE. Evaluation of the acaricide activity of extracts from eight species of medicinal plants on larvae of Hyalomma anatolicum (Acari: Ixodidae) ticks. 2024:102231.
32. Salman M, Abbas RZ, Israr M, Abbas A, Mehmood K, Khan MK, et al. Repellent and acaricidal activity of essential oils and their components against Rhipicephalus ticks in cattle. 2020;283:109178.
33. Aljaafari MN, Yap PSX, Seboussi R, Lai KS, Lim SHE. The potential use of essential oils as natural biocides against plant pathogens. Pesticides in the Natural Environment: Elsevier; 2022. p. 317-28.
34. Shah MA, Wani SH, Khan AAJJoFB, Nanoprocessing. Nanotechnology and insecticidal formulations. 2016;1(3):285-310.
35. Soudy BA, Allam SF, Hassan M, Khalil N, Youssef DAJEJoC. A new philosophy in fabrication pesticides (Abamectin) and Essential oils (Orange oil) into nano-form. 2021;64(7):3951-9.
36. Radwan IT, Khater HF, Mohammed SH, Khalil A, Farghali MA, Mahmoud MG, et al. Synthesis of eco-friendly layered double hydroxide and nanoemulsion for jasmine and peppermint oils and their larvicidal activities against Culex pipiens Linnaeus. 2024;14(1):6884.
37. Machado S, Pereira R, Sousa RMOJSoTTE. Nanobiopesticides: Are they the future of phytosanitary treatments in modern agriculture? 2023:166401.
38. Muntean D, Licker M, Alexa E, Popescu I, Jianu C, Buda V, et al. Evaluation of essential oil obtained from Mentha× piperita L. against multidrug-resistant strains. 2019:2905-14.
39. Harke M, Somkuwar A, Dubey S, Limsay R, Umap S, Sawarkar A, et al. Qualitative and quantitative phytochemical evaluation of ethanolic extract of Mentha pipperita (Linn.). 2021;10(5):996-1000.
40. Shetta A, Ali IH, Sharaf NS, Mamdouh WJIJoBM. Review of strategic methods for encapsulating essential oils into chitosan nanosystems and their applications. 2024:129212.
41. Bandaranayaka K, Dissanayaka U, Rajakaruna RJCJoS. Experimental aspects of life cycles of two hard tick species, Rhipicephalus (Boophilus) microplus and Hyalomma isaaci (Acari: Ixodidae), on New Zealand white rabbits. 2021;50(1):47-56.
42. Udobi MI. Time Since Blood-feeding (hunger) in Ixodes Ricinus Ticks: Measurement and Consequences: University of Bristol; 2023.
43. Njoroge PW. Effect of East Coast Fever Marikebuni Vaccine on Feeding and Reproductive Success of Rhipicephalus Appendiculatus: University of Nairobi; 2023.
44. Shakya M, Singh M, Gautam K, Jamra S, Agrawal V, Jayraw A, et al. Assessment of Antitick and Repellent Activity of Azadirachta indica Oil against Adults and Larvae Stages of Rhipicephalus microplus. 2024;2024.
45. Youssef AA, Abdelgawad AA, Abdel-Ghany HS, Abdelshafy SJEJoVS. Chemical Composition of the Essential Oil from Ruta chalepensis L. Growing Wild and its Acaricidal Activity Against the Cattle Tick Rhipicephalus annulatus (Acari: Ixodidae). 2024;55(4):955-64.
46. Jain P, Satapathy T, Pandey RKJEP. Rhipicephalus microplus (acari: Ixodidae): Clinical safety and potential control by topical application of cottonseed oil (Gossypium sp.) on cattle. 2020;219:108017.
47. Dzemo WD, Vudriko P, Ramatla T, Thekisoe OJP. Acaricide Resistance Development in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) Populations against Amitraz and Deltamethrin on Communal Farms of the King Sabata Dalindyebo Municipality, South Africa. 2023;12(7):875.
48. OECD. Test No. 404: Acute Dermal Irritation/Corrosion2015.
49. OECD. Test No. 402: Acute Dermal Toxicity2017.
50. Rajput S, Sharma P, Malviya RJRAiFN, Agriculture. Utilization of Herbal Components as Insecticidal and Repellent Effects. 2023;14(3):144-54.
51. Tourabi M, Nouioura G, Touijer H, Baghouz A, El Ghouizi A, Chebaibi M, et al. Antioxidant, Antimicrobial, and Insecticidal Properties of Chemically Characterized Essential Oils Extracted from Mentha longifolia: In Vitro and In Silico Analysis. 2023;12(21):3783.
52. Kalemba D, Synowiec AJM. Agrobiological interactions of essential oils of two menthol mints: Mentha piperita and Mentha arvensis. 2019;25(1):59.
53. Klafke GM, Thomas DB, Miller RJ, de León AAPJT, Diseases T-b. Efficacy of a water-based botanical acaricide formulation applied in portable spray box against the southern cattle tick, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae), infesting cattle. 2021;12(4):101721.
54. da Silva Lunguinho A, das Graças Cardoso M, Ferreira VRF, Konig IFM, Gonçalves RRP, Brandão RM, et al. Acaricidal and repellent activity of the essential oils of Backhousia citriodora, Callistemon viminalis and Cinnamodendron dinisii against Rhipicephalus spp. 2021;300:109594.
55. Kaya EC, Akdeniz M, Ugurlu P, Yener I, Firat M, Ertas AJJoEOBP. Content analysis, nanoparticle forming potential and acaricidal effects on Tick (Hyalomma marginatum) of essential oils from two common Thymbra species. 2024:1-22.
56. Castilho CV, Fantatto RR, Gaínza YA, Bizzo HR, Barbi NS, Leitão SG, et al. In vitro activity of the essential oil from Hesperozygis myrtoides on Rhipicephalus (Boophilus) microplus and Haemonchus contortus. 2017;27:70-6.
57. Bravo-Ramos J, Flores-Primo A, Paniagua-Vega D, Sánchez-Otero M, Cruz-Romero A, Romero-Salas DJE, et al. Acaricidal activity of the hexanic and hydroethanolic extracts of three medicinal plants against southern cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae). 2021;85(1):113-29.
58. Rai VK, Mishra N, Yadav KS, Yadav NP. Nanoemulsion as pharmaceutical carrier for dermal and transdermal drug delivery: Formulation development, stability issues, basic considerations and applications. Journal of controlled release. 2018;270:203-25.
59. Mohammadifar M, Aarabi MH, Aghighi F, Kazemi M, Vakili Z, Memarzadeh MR, et al. Anti-osteoarthritis potential of peppermint and rosemary essential oils in a nanoemulsion form: behavioral, biochemical, and histopathological evidence. BMC Complementary Medicine and Therapies. 2021;21(1):1-12.
60. Teshome K, Gebre-Mariam T, Asres K, Perry F, Engidawork E. Toxicity studies on dermal application of plant extract of Plumbago zeylanica used in Ethiopian traditional medicine. Journal of ethnopharmacology. 2008;117(2):236-48.