Therapeutic efficacy of polyherbal spray containing extracts of Andropogon citrates, Cymbopogon citratus, Ocimum sanctum, Pinus longifoia, Calotropis procera, Datura stramonium, Aegle marmelos, Ricinus communis, Azadirachta indica, Allium sativum, Carica papaya, Annona squamosa and Pongamia glabra was evaluated against tick infestation in buffaloes on the basis of decrease in tick count and improvement in haemato-biochemical parameters after treatment. Single application of polyherbal spray to 20 randomly selected tick infested buffaloes revealed significant reduction in mean tick count from 60.13±7.54 on day 0 to 21.8±4.26 on day 3, to 4.90±1.48 on day 7 while mean tick count was 14.72±2.74 on day 21 post treatment. Haematology revealed non-significant increase in Total Erythrocyte Count, Haemoglobin and Packed Cell Volume on day 21 post-treatment compared to pre-treatment values. No significant changes were observed in Total Leukocyte Count, Differential Leukocyte Count and Platelet Counts over a period of treatment. Also non-significant changes were observed in plasma glucose, total protein, albumin and globulin in treated buffaloes. The results of present study imply the clinical improvement in tick infested buffaloes treated with polyherbal spray and hence it could be probable preparation for use in buffaloes as an alternative to chemical acaricide.
Around 70 percent of Indian population is thriving on agriculture and allied sectors like livestock industry. The buffaloes are important milch animal reared in India whose population has shown increase of 3.19% (108.7 million) in 19th Livestock census compared to 105.3 million of 18th Census (Livestock Census, 2012). Buffaloes are frequently infested with numerous species of ticks, which are responsible for transmitting diseases like babesiosis, theileriosis and anaplasmosis. Also economic losses occur due to degradation of livestock health and production (Ghosh and Nagar, 2014). Tick infestation in livestock produce heavy economic losses to livestock industry due to blood sucking, irritating bites and transmission of various haemoprotozoan diseases. The estimated cost on control of ticks and tick borne diseases of animals in India was US $ 498.7 billion per annum (De Castro, 1997). Heavy tick infestation in wild buffaloes have shown significant negative association with innate immunity thereby signifying the importance of tick control for enhancing the immunocompetence of buffaloes against various infectious diseases (Anderson et al., 2013).
Organophosphates, pyrethroids, formamidine and macrocyclic lactones are the major classes of acaricidal chemicals used for tick control in livestock since long time (Davey et al., 2010; Rao et al., 2014). Intensive and indiscriminate use of acaricidal solutions for control of ectoparasites has culminated in emergence of resistance among different strains of tick populations (Bianchi et al., 2003; Pegram et al., 2000). Also the residues in milk and meat as well as the risk of environmental pollution are the other limitations for use of chemical acaricides in control of livestock ticks (Muhammad et al., 2008). To overcome the limitations of chemical acaricides, identification and validation of novel preparations with acaricidal potency for control and eradication of tick infestations in livestock is essential. Therefore present experiment was designed to evaluate the efficacy of polyherbal spray against tick infestation in riverine buffaloes to overcome the limitations of chemical acaricides.
Materials and Methods
Twenty riverine buffaloes with varying degree of tick infestation irrespective of age, sex and breed were randomly selected for the experiment. The ticks of Boophilus microplus (Rhipicephalus microplus) and Hyalomma anatolicum anatolicum were identified in experimental buffaloes.
Single application of polyherbal spray (*Clear Ticks- Rakesh Pharmaceutical, Kalol, Gandhinagar, Gujarat, India) containing of Andropogon citrates, Cymbopogon citratus, Ocimum sanctum, Pinus longifoia, Calotropis procera, Datura stramonium, Aegle marmelos, Ricinus communis, Azadirachta indica, Allium sativum, Carica papaya, Annona squamosa and Pongamia glabra was used in the present study. The spray was uniformly sprayed once over the body of buffaloes after measurement of tick count. Tick counts were measured from seven heavily infested areas over body viz., ear pinna, inguinal region, under tail, back region, wither, dewlap and neck and expressed as mean tick count. Efficacy of spray was evaluated based on reduction in unit tick count compared to pre-treatment counts. Tick counts were performed before application of spray as well as 3, 7 and 21 days after application of spray. Clinical signs like pruritus, scratching, rubbing, alopecia were recorded in tick infested buffaloes before treatment and 21 days post treatment. Blood samples were collected before treatment and 21 days post polyherbal spray treatment as per standard procedure and analysed for Hb, PCV, TEC, TLC, MCHC, MCV, MCH and DLC on fully automated haemoanalyzer (Diatron- Abacus Junior 3.11). Plasma samples were analysed for glucose, total proteins, albumin and globulin on semi-automated biochemistry analyser (CA 2005) using diagnostic kits (Autospan Diagnostics Ltd. Surat, India).
Values obtained from treatment group of buffaloes for tick count were analysed for mean, standard error and analysis of variance (ANOVA) at P < 0.05 significance level while haematology and biochemical parameters were analysed for mean, standard error and level of significance (P <0.05) with paired ‘t’ test by using SPSS software (Version: 16).
Various clinical signs observed in tick-infested cattle were itching, alopecia, redness, rubbing and pruritus in areas with higher density of ticks. Skin coat was coarse with severe hair fall in areas having heavy density of tick population. Treatment of buffaloes with polyherbal spray revealed significant reduction in clinical signs and hair fall from skin coat of affected buffaloes. Skin lustre was improved after 21 days due to significant reduction in the nuisance of ticks on body of buffaloes.
Acaricidal Potential of Polyherbal Spray against Tick Infestation in Cattle
Total tick count in study buffaloes before treatment and over a period of observations after treatment is given in Table 1. Mean tick count in study buffaloes before treatment was 60.13±07.54. Significant (P<0.05) reduction in total tick count was observed on day 3 (21.80±04.26), which persisted till day 7 (04.90±01.48) while non-significant increase in tick count to 14.72±02.74 was observed on day 21 after application of spray compared to day 0 tick count. Percent reduction in tick count was 63.74% on day 3, 91.85% on day 7 while 75.51% on day 21 of the trial period.
No adverse signs like irritation, itching, salivation etc. were observed in buffaloes treated with polyherbal spray. Application of polyherbal spray revealed reduction in the degree of fly infestation on the body of buffaloes and so reported to have additional fly repellent activity.
Effect of Polyherbal Spray on Haematological Parameters
Complete blood count was performed on blood samples collected before start of treatment and 21 days post treatment in study buffaloes. The values of complete blood count before treatment and 21 days after treatment are depicted in Table 2. Total Erythrocyte Count (6.17±0.29 vs. 7.16±0.45 x 1012/l), Haemoglobin (10.77±0.67 vs. 11.03±0.64 gm/dl ), Packed Cell Volume (32.78±2.10% vs 33.82±1.90%), Granulocyte Count (28.96±4.78% vs. 35.48±4.45%) and Platelet Count (149.2±16.90 vs. 166.1±14.90 x 109/l) revealed non-significant increase on 21 days post treatment while total leukocyte count (9.16±0.67 vs. 8.43±0.63 x x 109/l) and lymphocyte count (65.61±4.40% vs. 59.27±4.11%) revealed non-significant decrease on day 21 post treatment as compared to day 0 values.
Table 1: Mean± SE values of total tick count before and after treatment in buffaloes with R. microplus and H. anatolicum anatolicum infestation (n= 20).
|Sr. No.||Area of tick infestation||Total tick count|
|Before treatment||After treatment|
|Day 0||Day 3||Day 7||Day 21|
|Mean total tick count||60.13±07.54a||21.80±04.26bc
Table 2. Mean± SE values of haemato-biochemical values before and after treatment in buffaloes with R. microplus and H. anatolicum anatolicum infestation (n= 20).
|Sr. No.||Parameter||Trial period||‘t’ value|
|0 day||21st day|
|Total Erythrocyte Count (x 1012/l)||06.17±00.29||07.16±00.45||-2.036NS|
|Haemoglbin (gm/dl)||10.77±00.67||11.03±00.64||-1.452 NS|
|Packed Cell Volume (%)||32.78±02.10||33.82±01.90||-0.866 NS|
|Total Leukocyte Count (x 109/l)||09.16±00.67||08.43±00.63||1.853 NS|
|Granulocyte %||28.96±04.78||35.48±04.45||-1.768 NS|
|Monocytes %||05.43±01.39||05.25±01.27||0.253 NS|
|Lymphocytes %||65.61±04.40||59.27±04.11||1.764 NS|
|Platelets (x 109/l)||149.20±16.90||166.1±14.9||-1.627 NS|
|Total Proteins (gm/dl)||08.60±00.35||08.20±00.46||0.622NS|
Effect of Polyherbal Spray on Biochemical Parameters
Mean± SE values of plasma total proteins, albumin, globulin and glucose before application of spray and 21 days post treatment are depicted in Table 2. No significant difference (P< 0.05) was observed in values of plasma total proteins, albumin, globulin and glucose in buffaloes infested with tick on day 0 and 21 days post treatment.
Tick infestations in animals are quite severe causing harm through direct injuries or transmission of haemoprotozoan diseases and are responsible for economic losses to the tune of US $ 13.9 to 18.7 billion per year globally (De Castro, 1997). Indiscriminate use of the acaricides over a long period has culminated in the emergence of resistant tick population (Miller et al., 2005; Miller et al., 2007; Kumar et al., 2011; Sharma et al., 2012; Kumar et al., 2014 and Singh et al., 2014). To overcome the limitations as well as adverse effects of chemical acaricides, there is need for search of alternative strategies for tick control. Tick vaccines and phytoacaricides are the strategies which could be used as potential alternative to chemical acaricidess.
Phytoacaricides or herbal acaricides are beneficial in fighting resistance due to their rapid degradation and lack of persistence as well as bioaccumulation in the environment compared to synthetic chemical acaricides (Ghosh and Nagar, 2014). Tendonkeng et al (2005) and Cetin et al (2009) reported acaricidal activity of essential oil harvested from leaves and flowers of Ageratum houstonianum, Origannum onites and O. minutiflourum against R. (B.) annulatus and R. turancicus. Choudhary et al (2004) found acaricidal activity of tobacco leaves against R. haemaphysaloides ticks while Srivastav et al (2008) and Magadum et al (2009) reported good efficacy of ethanolic extracts of Azadirachta indica leaves, barks and seed (80%) and Anona squamosa seed (70.8%) against R. microplus ticks in vitro. Tripathi et al (2016) revealed promising results by thrice application of 1:20 diluted Keetguard liquid against B. microlus ticks in buffaloes.
In the present study polyherbal spray prepared from 13 different herbs might be having additive effect in significantly reducing tick count from 3 days to 7 day post treatment, while tick count was increased on day 21 post treatment. The increase in tick count on 21st day might be due to absence of persistent acaricidal effect and rapid degradation of the herbal formulations used in the study (Ghosh and Nagar, 2014).
Use of polyherbal spray against Rhipicephalus microplus and Hyalomma anatolicum anatolicum infestation in buffaloes revealed significant effect in reducing the tick count till day 7 post treatment with increasing trend towards day 21 post treatment. Polyherbal spray have shown good acaricidal potential till day 7 thereby reducing the nuisance of ticks to buffaloes and thus projecting polyherbal spray as potential alternative ectoparasiticide for controlling tick infestations in buffaloes.
Authors are thankful to Associate Dean, College of Veterinary and Animal Sciences, Udgir for kind support and also thankful to Rakesh Pharmaceuticals, Ahamadabad Gujarat, India for providing grants and polyherbal spray formulations for conducting study in riverine buffaloes.