5-Title: In-vitro and in-vivo evaluation of herbal extracts (garlic and papaya seed extract) against Rhipicephalus microplus in Western Maharashtra
Authors: BH Belsare, PD Pawar, GM Chigure, VS Dhaygude, SN Jadhav, VV Karande, VG Nimbalkar, NR Mahisekar, SR Patil, SH Belsare and LD Singla
Source: Ruminant Science (2025)-14(1):29-36.

Abstract

5. How to cite this manuscript: Belsare BH, Pawar PD, Chigure GM, Dhaygude VS, Jadhav SN, Karande VV, Nimbalkar VG, Mahisekar NR, Patil SR, Belsare SH and Singla LD (2025). In-vitro and in-vivo evaluation of herbal extracts (garlic and papaya seed extract) against Rhipicephalus microplus in Western Maharashtra. Ruminant Science 14(1):29-36.
Abstract
The present research aimed to evaluate the efficacy of herbal extracts-specifically garlic and papaya seed extracts—against Rhipicephalus microplus ticks infesting cattle across four distinct agro-climatic zones of Western Maharashtra, India. A total of 375 animals were screened for tick infestation during the study period from June to February, with 200 animals testing positive. The in-vitro efficacy of the herbal extracts was assessed using the Adult Immersion Test (AIT), Larval Packet Test (LPT), Larval Immersion Test (LIT), and Egg Hatch Assay (EHA). In-vivo trials were conducted by spraying both extracts on the animals and in their sheds. The in-vitro evaluations revealed that a 15% concentration of garlic extract led to a mortality rate of 66.66±6.66% in adult female ticks, with an LC50 value of 13.45% and an 86.44±0.7831% inhibition of oviposition. The 10% concentration of papaya seed extract exhibited a remarkable 86.66±6.66% mortality rate and an impressive 99.59±1.45% inhibition of oviposition, with an LC50 value of 7.09%. The LIT showed mortality rates of 72.62±6.09% for 5% garlic extract and 96.441±0.487% for 1% papaya seed extract. The 10% papaya seed extract demonstrated a significant 74.56% reduction in egg hatchability in the EHA, while the 15% garlic extract resulted in a notable 54.58% decrease. However, neither of the herbal extracts displayed significant efficacy against naturally tick-infested animals.
References
Aboelhadid SM, Kamel AA, Arafa WM and Shokier KA (2013). Effect of Allium sativum and Allium cepa oils on different stages of Boophilus annulatus. Parasitology Research 112:1883-1890.
Ajith KG, Fular A, Chigure G, Sharma AK, Nagar G, Souza FF, Bechara GH and Ghosh S (2019). Comparative impact of coumaphos, amitraz and plant extract of Ageratum conyzoides on the oogenesis of Rhipicephalus microplus. Ticks and Tick-Borne Diseases 10(5):1085-1095.
Bharkad GP, Palampalle HY, Narladkar BW, Bannalikar AS, Zende RJ and Ingole SD (2018). Effect of herbal acaricides on hatchability of eggs of deltamethrin and amitraz resistant cattle tick Rhipicephalus microplus. Journal of Entomology and Zoology Studies 6(5):629-633.
Bravo-Ramos JL, Flores-Primo A, Paniagua-Vega D, Sánchez-Otero MG, Cruz-Romero A and Romero-Salas D (2021). Acaricidal activity of the hexanic and hydroethanolic extracts of three medicinal plants against southern cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae). Experimental and Applied Acarology 85(1):113-129.
Dhamale Madhuri, Ravikumar RK, Ksheersagar Vivek Hindurao and Kumar Vipin (2017). Social construction of technology: An illustrative model for scaling up experimental wisdom of community in livestock welfare. Ruminant Science 6(1):119-123.
Finney DJ (1962). Probit Analysis. 2nd Edn, Cambridge University Press.
Ghosh Srikanta, Tiwari Shashi Shankar, Srivastava Sharad, Sharma Anil Kumar, Kumar Sachin, Ray DD and Rawat AKS (2013). Acaricidal properties of Ricinus communis leaf extracts against organophosphate and pyrethroids resistant Rhipicephalus (Boophilus) microplus. Veterinary Parasitology 192(1-3):259-267.
Heinrich M, Rimpler H and Antonio-Barrera N (1992). Indigenous phytotherapy of gastrointestinal disorders in a low land Mixe community (Oaxaca, Mexico): Ethnopharmacological evaluation. Journal of Ethnopharmacology 36:63-80.
Hitaishi VN, Lakshmanan Bindu, Jain Jose K, Emmanuel Seph Ross and Shameem H (2021). Molecular detection of Theileria spp. in ticks on goats in Kerala, South India. Ruminant Science 10(2):281-84.
Hurtado OJB and Giraldo-Ríos C (2018). Economic and health impact of the ticks in production animals. Ticks and Tick-borne Pathogens 9:1-9.
Jarial MS (2001). Toxic effect of garlic extracts on the eggs of Aedesaegypti (Diptera: Culicidae): A scanning electron microscopic study. Journal of Medical Entomology 38(3):446-450.
Katuri Ram Naresh, Das G and Singh Alok Kumar (2017). Distribution and prevalence of tick infestation on bovines in and around Jabalpur, Madhya Pradesh. Ruminant Science 6(1):63-67.
Kaur P, Juyal PD, Sharma A, Bal MS and Singla LD (2016). Seroprevalence of B. bigemina in dairy animals from low-lying regions of Punjab, India. Indian Journal of Animal Research 50(3):406-410.
Kumari Shimpi, Shekhar Pallav, Kumar Pankaj, Bachan Madhurendra, Kumar Ajit and Kumar Pankaj (2022). Molecular detection of Theileria annulata and Theileria orientalis in cattle and its prevalence in four agro-climatic zones of Bihar. Ruminant Science 11(2):267-274.
Meenal, Singh AP, Kachhawa JP and Nayak TC (2022). Studies on prevalence and risk factors associated with theileriosis in cattle calves in Bikaner. Ruminant Science 11(2):435-439.
Meriga B, Mopuri R and Murali Krishna T (2012). Insecticidal, antimicrobial and antioxidant activities of bulb extracts of Allium sativum. Asian Pacific Journal of Tropical Medicine 5(5):391-395.
Muraleedharan K, Sahadev A, Mallikarjuna G, Gopinathan N and Nagaraju S (2008). Screening of certain plants for controlling ticks in goats and sheep. Indian Journal of Animal Research 17:44-49.
Nasreen N, Niaz S, Khan A, Zaman MA, Ayaz S, Naeem H, Khan N and Elgorban AM (2020). The potential of Allium sativum and Cannabis sativa extracts for anti-tick activities against Rhipicephalus (Boophilus) microplus. Experimental and Applied Acarology 82:281-294.
Neelu S, Veer S and Shyma KP (2016). In vitro efficacy of commercial preparation of amitraz in Rhipicephalus (Boophilus) microplus ticks in North Gujarat, India. Ruminant Science 5(1):33-37.
Okolie NJC (2006). Larvicidal effects of paw paw (Carica papaya) aqueous extract on mosquito vectors. International Journal of Natural and Applied Sciences 2(4):417-420.
Parmar AB, Panchasara HH, Patel KB, Chauhan HC and Chandel BS (2018). Molecular detection of Babesia bigemina from blood of dairy cattle. Ruminant Science 7(2):287-288
Pendzhiev AM (2002). Proteolytic enzymes of papaya: Medicinal applications. Pharmaceutical Chemistry Journal 36(6):315-317.
Rani Bhawana, Yadav Kashyap, Yashavanth BS, Sanyal Aniket, Murugkar HV and Gandhale PN (2023). Evaluation of general health status and deworming as determinants in the CCHFV seroprevalence in animals in Madhya Pradesh. Ruminant Science 12(2):329-333.
Ravikumar RK, Kumar Vivek, Choudhary Hardev, Kinhekar Amol S and Kumar Vipin (2015). Efficacy of indigenous polyherbal ectoparasiticide formulation against hard tick infestation in cattle (Bos indicus). Ruminant Science 4(1):43-47.
Rawani A, Ghosh A, Laskar S and Chandra G (2012). Aliphatic amide from seeds of Carica papaya as mosquito larvicide, pupicide, adulticide, repellent, and smoke toxicant. Journal of Mosquito Research 2(2).
Saito KC, Bechara GH, Nunes ET, de Oliveira PR, Denardi SE and Mathias MIC (2005). Morphological, histological and ultrastructural studies of the ovary of the cattle tick Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae). Veterinary Parasitology 129(3-4):299-311.
Shyma KP, Singh V, Gupta JP and Pawar MM (2019). In vitro assessment of acaricidal activity of garlic cloves and papaya leaves xii against deltamethrin and cypermethrin susceptible Rhipicephalus (Boophilus) microplus. Ruminant Science 8(2):233-236.
Shyma KP, Gupta JP, Ghosh S, Patel KK, Singh V (2014). Acaricidal effect of herbal extracts against cattle tick Rhipicephalus (Boophilus) microplus using in vitro studies Parasitology Research 113(5):1919-26
Soulsby EJL (1965). Textbook of Veterinary Clinical Parasitology. FA Davis Co, Philadelphia.
Willadsen P (2006). Tick control: Thoughts on a research agenda. Veterinary Parasitology 138(1-2):161-168.