NAAS Score 2018


Declaration Format

Please download DeclarationForm and submit along with manuscript.


Free counters!

Previous Next

Comparative Efficacy of Herbal (Lemon Grass Oil and Litsea Grass oil) and Synthetic (Deltamithrin, Fipronil, Amitraz) Agents against Rhipicephalus microplus Infestation in Mithun (Bos frontalis)

Prakash Ranjan Dutta Jayanta Kumar Chamuah Akhilesh Kumar Runjun Dowarah Meraj Haider Khan
Vol 8(11),221-228

Ticks are important ectoparasites which results in economic loss in livestock industry. Therefore, the present study was undertaken with aim to study the comparative efficacy of some herbal extracts i.e. oil of Lemon grass (Cymbopogon citratus) and Litsea grass (Litsea cubeba) and three synthetic acaricides viz. Deltamethrin, Fipronil, and Amritraz against the Rhipicephalus microplus tick infestation in mithun. Among the chemical drugs, Fipronil showed 100% efficacy followed by Amitraz (98.38%) and Deltamithrin (93.28%). The lemon grass oil on 12 days of application resulted in very high efficacy (98.00 %) comparable to synthetic acaricides (Fipronil, Amitraz and Deltamithrin), however, Litsea grass oil did not showed significant efficacy on day 12 post application. However, the Adult Immersion Test (AIT) showed dose dependent acaricidal efficacy for Deltamethrin, Fipronil and Amritraz at final concentration of undiluted, 0.40%, 0.20 %, 0.10 % and 0.05% while among the herbal acaricides in 100 % concentration, lemon grass oil showed comparable efficacy with deltamethrin (12.5 W/V) while Listea grass oil did not showed better effects. Author concludes that lemon grass oil has potential acaricidal activity and may be recommended for holistic management of nymphal stages of tick infestation and can be proved as an alternative to synthetic acaricides without adversely affecting health and environment.

Keywords : Herbal Ticks Efficacy Ectoparasites Mithun Acaricidal

Ticks are among the most competent and versatile vectors of pathogens and are second to mosquitoes as vectors of a number of human pathogens, like viruses, bacteria, rickettsia, spirochetes and the most important vector of pathogens affecting cattle worldwide (Ghosh and Nagar, 2014 and Kakati et al., 2015). Ticks represent a serious problem worldwide among livestocks leading to huge losses by transmitting diseases. Rhipicephalus microplus also referred as Asian Blue Tick that parasitises a variety of livestock species in tropical and subtropical regions (Klafke et al., 2006). Initially, this tick was geographically limited to Asian continent later it has been reported from Mexico, Central and South America, Africa, Madagascar, Australia, and Taiwan (Labruna et al., 2009). The  R. microplus mainly infests cattle, deer and buffalo, but it can also be found on horses, goats, sheep, donkeys, dogs, pigs and some wild mammals (Kahn and Line, 2003). R. microplus has been found potential vectors and reservoirs Babesia bovis, Anaplasma phagocytophilum, Anaplasma ovis, Ehrlichia canis, Ehrlichia ewingii and Ehrlichia muris (Matysiak et al., 2016).

Mithun (Bos frontalis), unique ruminant species geographically limited to North-Eastern Hilly region of India and particularly to the states of Arunachal Pradesh, Nagaland, Manipur and Mizoram. Traditionally mithun being reared under forest ecosystem make the animal prone to ecotoparatic infestation. However, the limited reports of different tick infestation are available in mithun (Chamuah et al., 2012; Chamuah et al., 2016). Management of tick infestation in mithun with different acaricides has been attempted by various workers from time to time (Chamuah et al., 2014). The emergence of resistance is a constant challenge for control of R. microplus (Morel et al., 2017) and development of new acaricides is a long and expensive process, which reinforces the need for alternative approaches to control ticks infestations (Graf et al., 2004). In past few years, the studies on herbal acaricides have gain momentum worldwide in attempt to overcome the increasing resistant to chemical acaricides (Magadum et al., 2009; Chagas et al., 2011).

Keeping in view the rapid development of resistance, the present study was designed with aim to investigate the acaricidal in-vitro efficacy of Lemon grass oil and Litsea grass oil in comparison to commercial synthetic acaricides against R. microplus infestation in mithun.

Materials and Methods

A total of 24 numbers of R. microplus infested mithuns were divided into six groups. The groupings were done randomly irrespective of age and sex of the animals. Each group was treated separately with Deltrin (deltamethrin emulsifiable concentrate-1.25% W/V, ARIANS BIOPHARMA), Nayflee (Fipronil 0.25% W/V, Intas Pharmaceutical Ltd., Ahmedabad), Extick (Amritraz dip concentrate liquid 12.5% W/V Vet Mankind, New Delhi) in undiluted form. Lemon grass (Cymbopogon citratus) and Litsea (Litsea cubeba) were collected locally from Medziphema, Dimapur, Nagaland and essential oil were prepared by steam distillation (Boukhatem et al., 2014).  Four tick infested animals were maintained without any treatment as the control group. The herbal extracts were prepared as per conventional procedure. Efficacy of various treatments was analyzed at 0th, 3rd, 6th, 9th and 12th days of treatment. The efficacy of all the drugs was expressed by per cent reduction of nymphal stages of ticks per square inch body surface area of the affected animals (Ong’are et al. 1983).

Adult Immersion Test

Briefly, the Adult Immersion Test (AIT) was conducted as per the protocol described by Drummond et al., 1973.  A total of 4 ticks for each groups were immersed in pertidish having 10mL of chemical acaricides (Deltamethrin, Fipronil, Amitraz) in different concentrations of namely, undiluted, 0.40 %, 0.20 %, 0.10 % and 0.05 %. The ticks were exposed to 100 % concentration of Litsea grass oil and Lemon grass oil. The control group ticks were immersed in water. The ticks were observed for cessation of pedal reflex on pricking of needle and time of cessation of movement was recorded.

Statistical Analysis

The experimental data generated were analyzed by adopting repeated measures GLM procedure using the statistical software program SPSS (version 17.0 for Windows; SPSS, Chicago, III., U.S.A.). A one-way analysis of variance (ANOVA) was used for comparison of means according to Duncan’s multiple range tests (Duncan, 1995). The effects were considered to be significant at 5% level.


The findings of the study of efficacy of certain herbal and commercial synthetic acaricides against R. microplus infestation in mithun has been presented in Table 1.

Table 1: Efficacy of acaricides against tick infestation in mithun

Groups Treated with No. of Nymph/Tick Present on Day of Treatment
0 day 3rd day 6th day 9th day 12th day
I Deltamithrin 20.25 ab ±1.45 11.5 c ±1.50 (43.20%) 3.75 ab ±0.75 (81.48%) 1.75 a ±0.48 (91.35%) 1.25 a ±0.48 (93.82%)
II Fipronil (Nayflee) 16.5 a ±1.56 0.00 a ±0.00 0.0 a ±0.00 0.0 a ±0.0 0.0 a ±0.0
III Amitraz (Extick) 15.5 a ±3.22 5.50 b ± 1.56 (64.51%) 1.0 a ±0.41 (93.54%) 0.25 a ±0.25 (98.38%) 0.25 a ±0.29 98.38%
IV Lemon grass oil 25.5 b ±2.32 13.5 cd ±1.71 (47.05%) 6.5±0.87b (74.5%) 1.75±1.18a (93.13%) 0.5±0.50 a (98%)
V Listea oil 20.25 ab ±2.56 16.75 de ±2.75 (17.25%) 13.50 c ±2.53 (33.33%) 13.25 b±2.66


13.0 b ±1.78 (35.8%)
VI Control group 21.75 ab ±0.75 21.5 e ±1.32 20.75 d ±1.25 21.0 c ±0.91 21.25 c ±0.63
P value 0.045 <0.001 <0.001 <0.001 <0.001

abcde superscript differ significantly (P<0.05) between the rows


The Fipronil (Nayflee) have shown 100 % efficacy on the third day of application and onwards whereas Amritraz (Extick) Deltrin and lemon grass oil have shown significant increase in efficacy on day 12 in comparison to day 0. The Litsea extract did not show significant efficacy in terms of reduction of nymphal stages of ticks on day 12 in comparison to day 0. No variation in the control group animals in terms of reduction of nymphal stages of tick was observed in the course of the study. The findings of AIT revealed the dose dependent increase in acaricidal efficacy (Table 2). Among, undiluted commercial preparations, deltamethrin showed highest efficacy followed by lemon grass oil, Amitraz and Fipronil.

Table 2: Dose dependent response of Adult Immersion Test (AIT) to various commercial and herbal acaricidal preparations against R. microplus

Final Concentration Deltamethrin 1.25% W/V Amritraj 12.5% W/V Fipronil 0.25% W/V Litsea grass oil (100 %) Lemon grass oil (100 %)
      Undiluted     4 minutes 29 minutes 36 minutes 68 minutes 7 minutes
0.40% 70 minutes 67 minutes 72 minutes
0.20% 83 minutes 94 minutes 92 minutes
0.10% 87 minutes 113 minutes 99 minutes
0.05% 91 minutes 142 minutes 115 minutes


Ticks represent a serious problem worldwide among livestocks leading to huge losses by transmitting diseases. The emergence of resistance to synthetic acaricidal agents are a constant challenge for control of R. microplus. In present study for per cent reduction of nymphal stages of ticks per square inch body surface area of the affected animals, Fipronil showed 100% efficacy followed by Amitraz (98.38%) and Deltamithrin (93.28%). However, on AIT commercial acaricides have shown the dose dependent acaricidal efficacy which was recorded highest in Deltamethrin followed by Amritraz and Fipronil at final concentration of undiluted, 0.40%, 0.20 %, 0.10 % and 0.05%. In other studies on the efficacies of cypermethrin and deltamethrin on the R. microplus populations varied from 48.33% to 70.5% and 61.22% to 76.84%, respectively (Brito et al., 2011). Another study using cis-cypermethrin (0.01%) + DDVP (0.11%), coumaphos (0.05%), deltamethrin (0.0025%), amitraz (0.025%) and cypermethrin (0.015%) on R. microplus showed  that  eggs treated with high cis-cypermethrin and cypermethrin has eclosion inhibitions of 72.1% and 67.3%, respectively, whereas coumaphos was much less effective (only 11.7%). Except for coumaphos, which resulted in 63.3% and 80.0% mortality of nymphs and unfed females, respectively, all compounds tested killed 100% of all tick stages to which they were exposed (Bicalho et al., 2001).

However the  studies suggests that synthetic acaricidal agents fail to overcome the problem and  selective treatment and culling do not represent feasible methods to control R. microplus infestation on cattle (Morel et al., 2017). The resistance of R. microplus to organophosphates (chlorpyrifos, coumaphos and diazinon), pyrethroids (flumethrin, deltamethrin and cypermethrin), phenylpyrazole (fipronil), amitraz and ivermectin has been documented (Sharma et al., 2012; Rodríguez-Vivas et al., 2014, and Shyma et al., 2015). Herbal acaricides may be a potential substitute for synthetic acaricides currently used against tick. The present study showed that lemon grass oil on 12 days of application resulted in very high efficacy comparable to synthetic acaricides (Fipronil, Amitraz and Deltamithrin). Further, the lemon grass oil has also showed the comparable acaricidal efficacy on AIT. In various studies the lemon grass oil has been proved to have anti-bacterial, anti-inflammatory and anti-fungal properties (Naik et al., 2010; Boukhatem et al., 2014), however, the acaricidal efficacy has not been studied.

In a study of efficacy of extracts from leaf, bark, and seed of Azadirachta indica, leaf and seed of Prunus persica, bark of Mangifera indica, and leaf of Psidium guajava against Boophilus microplus, the A. indica seed extract have shown 80 % efficacy after 5 h of treatment and caused the significant reduction (P < 0.01) in the reproductive index of ticks in comparison to control. The efficacy of the neem seed extracts was also found comparable with the commonly used synthetic pyrethroids against B. microplus (Srivastava et al., 2008). Pirali-Kheirabadi and Teixeira da Silva (2010) studied Lavandula angustifolia essential oil as a novel and promising natural candidate for tick Rhiphicephalus annulatus control. Pirali-Kheirabadi and Teixeira da Silva (2011) also reported In vitro efficacy of the acaricidal properties of Artemisia annua and Zataria multiflora essential oils to control cattle ticks. Chagas et al. (2011) reported In vitro efficacy of Artemisia annua extract against Rhiphicephalus microplus. In vitro efficacy of ethanolic extracts of Ageratum conyzoides and Artemisia absinthium was also assessed on Rhipicephalus microplus using adult immersion test (AIT). The A. absinthium was to have better acaricidal properties than A. conyzoides againts R. microplus (Praveen et al., 2014).

The 100% acaricidal activity of Annona squamosa and 60% activity of neem oil against ticks of genera Boophilus microplus and R. haemaphysalis were reported by Kalakumar et al. (2000). However, Magadum et al. (2009) had reported 70.8% efficacy of A. squamosa against R. microplus during In-vitro trial. While, Kumar et al. (2005) had reported that a 50:50 mixture spray of neem + karanj oil was effective on 97.95 per cent of ticks from natural infection in cows on day 15 post application suggesting that formulation is safe alternative to control ticks in field conditions. In a comprehensive study by Ghosh et al. (2013), 95% ethanolic extract of Ricinus communis extract significantly affected the mortality rate of ticks with an additional effect on reproductive physiology by inhibiting oviposition. Chamuah et al. (2014) has also recorded efficacy of crude extract of neem at high concentration and crude extract of tobacco at low concentration against R. microplus in naturally infested mithun. The leaves of tobacco (Nicotina tabacum) was also found effective against R. haemaphysaloides (Choudhury et al., 2004).



The author concludes that lemon grass oil could be used as a potential substitute to synthetic acaricides due to natural origin, safe and environment friendly. Further, the repetitive and long term application of lemon grass oil did not pose any problem to development resistance which occurs commonly with synthetic agents.


The authors duly acknowledge the  Director, ICAR-National Research Centre on Mithun , Medziphema, Nagaland for providing the facilities and help during the period of the study.


  1. Bicalho KA, Ferreira F, Borges and Ribeiro MFB (2001). In vitro evaluation of the effects of some acaricides on life stages of Rhipicephalus sanguineus. Brazilian Journal of Veterinary and Animal Science 53 (5): 548-552.
  2. Boukhatem MN, Ferhat MA, Kameli, A, Saidi, F. and Kebir HT. 2014. Lemon grass (Cymbopogon citratus) essential oil as a potent anti-inflammatory and antifungal drugs. Libyan Journal of Medicine 9: 25431.
  3. Brito LG, Barbieri, FS, Rocha RB, Oliveira MCS and Ribeiro ES (2011). Evaluation of the Efficacy of Acaricides Used to Control the Cattle Tick, Rhipicephalus microplus, in Dairy Herds Raised in the Brazilian Southwestern Amazon Veterinary Medicine International, Article ID 806093, 6 pages doi:10.4061/2011/806093.
  4. Chagas ACS, Georgetti CS, de Carvalho CO, de Sena Oliveira MC, Rodrigues RA, Foglio MA, and de Magalhães PM. 2011. In vitro activity of Artemisia annua L (Asteraceae) extracts against Rhipicephalus (Boophilus) microplus. Revista Brasileira de Parasitologia Veterinaria 20 (1): 31–35.
  5. Chamuah JK,  Dutta PR, Prakash V, Raina OK, Sakhrie A,  Borkotoky D,  Perumal P, Neog    and Rajkhowa  C. 2014. Comparative efficacy of some plant extracts on Rhipicephalus (Boophilus) microplus infestation on mithun (Bos frontalis) in the north east. Journal of Veterinary Parasitology 27:5-7.
  6. Chamuah JK, Singh V, Dutta PR, Khate K, Mech A. and Rajkhowa C. 2012. Tick infestation in mithun. Indian Veterinary Journal  89: 140
  7. Chamuah JK, Lama S, Dutta PR, Raina OK. and Khan  2016. Molecular identification of Ixodid ticks of mithun (Bos frontalis) from Nagaland. Indian Journal of Animal Science 86:762–763.
  8. Choudhary RK, Vasanthi C, Latha BR. and John L. (2004). In vitro effect Nicotiana tobacum aqueous extract on Rhipicephalus haemaphysaloides Indian Journal of Animal Science. 74: 730-731.
  9. Drummond RO, Ernst SE, Trevino JL, Gladney WJ, and Graham, OH. (1973). Boophilus annulatus and microplus: laboratory tests of insecticides. Journal of Economic Entomology 66 (1): 130–133.
  10. Duncan DB. 1995. Multiple range and multiple F-test. Biometrics II: 1-42.
  11. Ghosh S. and Nagar G. (2014). Problem of ticks and tick-borne diseases in India with special emphasis on progress in tick control research: A review. Journal of Vector Borne Diseases 259–270.
  12. Ghosh S, Tiwari SS, Srivastava S, Sharma AK, Kumar S and Ray DD. Acaricidal properties of Ricinus communis leaf extracts against organophosphates and pyrethroids resistant Rhipicephalus (Boophilus) microplus. Veterinary Parasitology 192: 259-67.
  13. Graf JF, Gogolewski R, Leach-Bing N, Sabatini GA, Molento MB, Bordin EL. and Arantes GJ. 2004. Tick control: an industry point of view. Parasitology 129:S427–S442.
  14. Kahn CM. and Line S. 2003. The Merck veterinary manual [online]. Whitehouse Station, NJ: Merck and Co.
  15. Kakati P, Sarmah PC, Ray D, Bhattacharjee K, Sharma RK, Barkalita LM, Sarma DK, Baishya BC, Borah P. and Stanley B. 2015. Emergence of oriental theileriosis in cattle and its transmission through Rhipicephalus (Boophilus) microplus in Assam, India. Veterinary World 8(9): 1099-1104.
  16. Kalakumar B, Kumar HSA, Kumar KBA and Reddy, K. S. 2000. Evaluation of custard seed oil and neem oil as acaricides. Journal of Veterinary Parasitology 14:171-172.
  17. Klafke GM, Sabatini GA, Albuquerque TA, Martins JR, Kemp, DH, Miller RJ. and Schumaker TTS. 2006. Larval immersion tests with ivermectin in populations of the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) from State of Sao Paulo, Brazil. Veterinary Parasitology 142: 386–390.
  18. Kumar S, Prasad KD, Deb AR, and Kumar A. 2005. Efficacy of ivermectin and neem with Karanj oil against natural Boophilus microplus infestation in cattle. Journal of Veterinary Parasitology 19: 59-60
  19. Labruna MB, Naranjo V, Mangold AJ, Thompson C, Estrada- Pena A, Guglielmone AA, Jongejan F. and de la Fuente J. 2009. Allopatric speciation in ticks: genetic and reproductive divergence between geographic strains of Rhipicephalus (Boophilus) microplus. BMC Evolutionary Biology 9: 46.
  20. Magadum S, Mondal DB and Ghosh S. 2009. Comparative efficacy of Annona squamosa and Azadirachta indica extracts against Boophilus microplus, Izatnagar isolate. Parasitology Research 4: 105: 1085-1091.
  21. Matysiak A, Dudko P, Dudek K,  Dudek M, Junkuszew A. and  Tryjanowski   2016. The occurrence of pathogens in Rhipicephalus microplus ticks from cattle in Madagascar. Veterinarni Medicina 61 (9): 516–523.
  22. Morel N, Signorini ML, Mangold AJ, Guglielmone AA. and Nava S. 2017. Strategic control of Rhipicephalus (Boophilus) microplus infestation on beef cattle grazed in Panicum maximum grasses in a subtropical semi-arid region of Argentina. Preventive Veterinary Medicine 144 (1): 179-183.
  23. Naik, M. I., Fomda BA, Jaykumar E. and Bhat JA. 2010. Antibacterial activity of lemongrass (Cymbopogon citratus) oil against some selected pathogenic bacterias. Asian Pacific Journal of Tropical Medicine 535-538.
  24. Ong’are JO, Rinkanya FG, Mwangi EN. and Kiundi JI. 1983. Evaluation of certain ixodicides for the control of Rhipicephalus appendiculatus using a tick-infested paddock. The Kenya Veterinarian 7: 3-6.
  25. Parveen S, Godara R, Katoch R, Yadav A, Verma PK, Katoch M. and Singh  2014.   In Vitro Evaluation of Ethanolic Extracts of Ageratum conyzoides and Artemisia absinthium against Cattle Tick, Rhipicephalus microplus. Scientific World Journal. Article ID 858973, 6 pages
  26. Pirali-Kheirabadi KH. and Teixeira da Silva JA. 2010. Lavandula angustifolia essential oil as a novel and promising natural candidate for tick (Rhipicephalus (Boophilus) annulatus) control. Experimental Parasitology 126(2):184-186.
  27. Pirali-Kheirabadi KH. and Teixeira da Silva JA. (2011). In-Vitro Assessment of the Acaricidal Properties of Artemisia annua and Zataria multiflora Essential Oils to Control Cattle Ticks. Iranian Journal of Parasitology 6 (1): 58-65.
  28. Rodríguez-Vivas RI, Pérez-Cogollo LC, Rosado-Aguilar JA, Ojeda-Chi MM, Trinidad-Martinez I, Miller RJ, Li AY,  de León AP, Guerrero F,  and  Klafke G. 2014.  Rhipicephalus (Boophilus) microplus resistant to acaricides and ivermectin in cattle farms of Mexico. Brazilian Journal of Veterinary Parasitology 23 (2): 113-122.
  29. Sharma AK, Kumar R, Kumar S, Nagar G, Singh NK, Rawat SS, Dhakad ML, Rawat AK, Ray DD. and Ghosh S. 2012. Deltamethrin and cypermethrin resistance status of Rhipicephalus (Boophilus) microplus collected from six agro-climatic regions of India. Veterinary Parasitology 188(3-4):337-345.
  30. Shyma KP, Gupta JP, Singh V. and Patel KK. 2015. In Vitro Detection of Acaricidal Resistance Status of Rhipicephalus (Boophilus) microplus against Commercial Preparation of Deltamethrin, Flumethrin, and Fipronil from North Gujarat, India. Journal of Parasitology Research., Article ID 506586, 7 pages.
  31. Snedecor GW. and Cochran WG. 1989. Statistical Methods. Oxford and IBH Publishing Co., New Delhi.
  32. Srivastava R, Ghosh S, Mandal DB, Azhahianambi P, Singhal PS. 2008. Efficacy of Azadirachta indica extracts against Boophilus microplus. Parasitology Research 104:  149-153.
Abstract Read : 52 Downloads : 26
Previous Next