NAAS Score – 4.31

Free counters!


Previous Next

Efficacy of Supplementing Ashwgandha Extract (Withania Somnifera) on Immunity and Serum Biochemistry in Broilers during Summer

K. Prashanth Kumar V. Ravinder Reddy M. Gnana Prakash
Vol 8(9), 158-164

The objective of the present study was to determine the effects of different levels of Ashwagandha (Withania somnifera) extract (WSE) on immunity and serum biochemical parameters of broiler chicken reared during hot summer. The average minimum and maximum room temperatures recorded from 0-42 day were 36.8oC±0.25 – 39.7oC±0.25, respectively, with relative humidity ranging from 47 – 74%. Day old broiler chicks (n=160) were randomly allotted to 4 dietary groups with eight replicates of five birds each. The four dietary groups were Positive control (PC) with vit-E (70mg/kg) +Se (0.15mg/kg), Negative control (NC), WSE50 and WSE100 groups supplemented with WSE 50mg/kg and 100mg/kg, respectively. Serum cholesterol levels were significantly (P<0.05) reduced in WSE50 and WSE100 groups, while serum protein levels were unaffected (P>0.05) by supplementation of WSE. Humoral immune response in terms of NDV (Newcastle disease virus) titer and cell mediated immunity in terms of cutaneous basophilic hypersensitivity (CBH) were not significantly influenced by supplementation of WSE in broiler diets.

Keywords : Ashwagandha Extract Cell Mediated Immunity Humoral Immunity Serum Cholesterol Serum Protein

Stress is defined as any biological response developed by an organism when its homeostasis is disrupted (Virden and Kidd 2009). High ambient temperature is considered as a potent climatic stressor causing impaired antioxidant status in poultry (White head and Keller, 2003). Heat stress can result in oxidative stress in birds which has been reported to decrease production performance and immunity hence results in pathogenesis of various diseases (Mujahid et al., 2005). Increase in stress induces sympatho-adrenal activity which further leads to release of corticosteroids, hormones that increase protein and lipid catabolism in turn elevating plasma cholesterol concentration (Sahin et al., 2004). Hypercholesteremia is one of the risk factors for coronary artery disease (CAD) (Gambhir et al., 2000). Heat stress decreases the lymphoid organ weights, reduces the levels of total circulating antibodies (Bartlett and Smith, 2003) and causes damage to T-lymhocytes and B-lymphocytes by lipid peroxidation. Dietary modifications are the most practical ways to alleviate heat stress in broilers by addition of anti-oxidants like Vit-E, Vit-C and Selenium (Sahin et al., 2004). Supplementation of medicinal plants as ingredients for broilers is increasing due to its growth promoting effects, immunomodulatory and hypolipidemic effects. Withania somnifera also known as ashwagandha has adoptogenic, immunomodulatory, hypoliplidemic and anti-stress activity (Davis and Kuttan, 2000). The main constituents of ashwagandha are withanolides, flavonoids and polyphenolic compounds (Uddin et al., 2012). So the present study was aimed to determine the effects of different levels of ashwagandha on immunity and serum biochemical parameters in broilers reared during hot summer.

Materials and Methods

Day old cobb broiler chicks (n=160) were randomly assigned to 32 replicates under 4 dietary treatment groups each with 5 chicks and kept in a closed, ventilated, wire-floor battery caged broiler house under uniform conditions of temperature, humidity, and ventilation.

Table 1. Ingredient and nutrient composition of basal diet

Ingredients (%) Starter (0-21d) Finisher (22-42d)
Maize 56.9 61.31
Soya bean meal 35.43 30.6
Vegetable oil 3.355 4.287
Salt 0.423 0.422
Dicalcium phosphate 1.602 1.542
Limestone powder 1.343 1.177
DL-methionine 0.278 0.217
L-Lysine 0.241 0.076
AB2D3K* 0.015 0.015
B-complex** 0.015 0.015
Trace mineral mixture*** 0.1 0.1
Nutrient Composition
ME (kcal/kg) 3050 3150
Crude protein (%) 21.5 19.5
Lysine (%) 1.25 0.98
Methionine (%) 0.56 0.48
Calcium (%) 0.9 0.82
Available phosphorous (%) 0.42 0.4

*AB2D3K provided per kg diet: Vitamin A 20000 IU, Vitamin B2 25 mg ,Vitamin D3 3000IU, Vitamin K 2mg. **Riboflavin 25mg, Vitamin B1 1mg, Vitamin B6 2mg, Vitamin B12 40mg, and Niacin 15mg. ***Trace mineral provided per kg diet: Manganese 120mg, Zinc 80mg, Iron 25mg, Copper 10mg, Iodine 1mg.

Birds were vaccinated against Newcastle (7th and 28th d) and infectious Bursal disease (15th d) as per the standard vaccination schedule. Maize and soybean meal-based diets were prepared to contain 3050 and 3,150 kcal ME/kg during the starter (0 to 21 d) and finisher (22 to 42 d of age) phases, with respective crude protein contents of 21.5 and 19.5 g/100 g feed (Table 1).

The experimental treatments were as follows: Positive control (PC) (basal diet +Vit-E 70mg/kg + Se 0.15 mg/kg), Negative control (NC)-basal diet, WSE50 (basal diet + 50 mg WS extract/kg diet) and WS100 (Basal diet + 100mg WS extract/kg diet). Water and mash feed were provided adlibitum throughout 42 days trial period. The temperature and relative humidity of shed during trial period were 36.8oC±0.25 – 39.7oC±0.25 and 47 – 74% respectively.


The humoral immunity was determined in birds by measuring antibody titre to Newcastle disease (ND) vaccine (antibody production against ND virus). At 21st and 42nd days of age blood was collected and serum was separated. Haemagglutination inhibition (HI) activity of serum was estimated and the antibody titers (log2) were measured following the standard procedure (Wegmann and Smithies., 1966). The cell mediated immune (CMI) response was assessed by measuring cutaneous basophilic hypersensitivity (CBH) to phytohaemagglutinin phosphate (PHA-P). On 40th day of experiment, one bird per replicate was injected with 100 μg of PHA-P suspended in 0.1 ml of phosphate buffer saline (PBS) into the web between third and fourth inter-digital space of right foot, while the left web (control) was injected with 0.1 ml of PBS. The web thickness of both feet was measured by micrometer after 24 h of injection and CBH was calculated by method of Edelman et al., 1986.

Serum Bio-Chemistry

On 41st day of experiment, blood collected from one bird per replicate into a clean sterilized tube and kept in a slanted position at room temperature to facilitate separation of serum. Serum protein and cholesterol were determined by using commercially available diagnostic kits (Erba Mannheim Chemicals).

Statistical Analysis

The data was analysed by using SPSS 15th version.

Results and Discussion

The mean (log2) antibody response to ND vaccine at 3rd and 6th week of age and cell mediated immune response at 41st day were not statistically significant (P>0.05). These results are similar to the findings of Mushtaq et al. (2012) who supplemented ashwagandha in drinking water (20g/litre). On the contrary Akotkar et al. (2007) reported that feeding 1.25% ashwagandha significantly improved HI titer and CMI in broilers. Similarly Ansari et al. (2013) and Vasanthakumar et al. (2015) reported improved ND titer in groups fed WSE at 0.15% and ashwagandha root powder at 2.5% – 5% levels in broiler diets respectively. Improved HI and CMI was also reported by feeding ashwgandha in broiler diets (Davis and Kuttan, 2000; Sujatha et al., 2010; and Singh et al., 2016). Improved immune parameters may be due to antimicrobial and immunomodulatory effects. Immunomodulatory effects of ashwagandha due to presence of glycowithanolides (Ghosal et al., 1989), which stimulates the production, phagocytic activity of circulating macrophages (Malik et al., 2007), stimulates bone marrow (Davis and Kuttan, 2000) and increases antibody producing lymphocytes. Decreased immune organ (spleen) weight corroborates the decreased immune response (HI & CMI) in the present study, which might be due to higher temperatures (36.8oC±0.25 – 39.7oC±0.25) during trial period. High temperatures are reported to reduce lymphoid organ weights (Hassan et al., 2007) and decreased circulating macrophages and antibodies (Smith, 2003; Mashaly et al., 2004; Niu et al., 2009). This reduction could be indirectly due to stimulation of hypothalamic pituitary adrenal (HPA) axis which results in release of corticosterone that inhibits antibody production (Gross et al., 1992; Star et al., 2008).

Table 2: Effect of ashwagandha extract on humoral immunity and cell mediated immunity in broiler chicken (1-42 d)

Treatment NDV titers (log2) *PHA-P Response (Thickness Index)
3rd wk 6th wk 6th week
PC 3.25 3.75 164.1
NC 3.5 2.75 144.3
WSE50 3.25 3.25 150.3
WSE100 3.38 3.75 162.6
SEM 0.133 0.132 2.355
P-Value 0.678 0.103 0.452

Means bearing different superscripts in a column differ significantly (P<0.05).

Table 3: Effect of ashwagandha extract on spleen weights, serum cholesterol and serum total protein levels in broiler chicken (1-42 d)  

Treatment Cholesterol (mg/dl) Total Protein (g/dl) Spleen (% live weight)
PC 148.6ab 3.26 0.09
NC 164.4b 2.85 0.08
WSE50 146.2a 3.07 0.09
WSE100 132.8a 3.67 0.07
SEM 3.558 0.127 0.007
P-Value 0.009 0.123 0.774

Means bearing different superscripts in a column differ significantly (P<0.05).

Serum cholesterol concentrations were significantly (P<0.05) decreased in groups supplemented with WSE, while serum protein levels were unaffected (P>0.05). WSE100 group recorded low level of serum cholesterol compared to NC group and comparable to WSE50 and PC group. Increase in stress induces sympatho-adrenal activity which further leads to protein and lipid catabolism in turn elevating plasma cholesterol concentration (Sahin et al., 2004). Low cholesterol in PC compared to NC may be due to Se, due to inhibitory effect on 3-hydroxy-3-methylglutaryl-coA (HMG-CoA) reductase, a key enzyme in cholesterol biosynthesis (Nassir et al., 1997). By quenching free radicals Vit-E reduces the effects of oxidative stress there by reduces corticosteroids secretion (Sahin et al., 2002). Similar results were found by Habibian et al. (2014) by feeding Vit-E at 250 mg/kg & Se at 0.5-1 mg /kg diet. Ansari et al. (2013) reported decreased cholesterol levels by feeding ashwagandha 2.5% & 5.0%. Similarly, Sujatha et al. (2010) reported reduced cholesterol levels by supplementation of polyherbal mix at 0.1% level, reduced cholesterol levels due to hypolipidaemic and hypocholesterolaemic effect of E. officinalis in polyherbal mixture (Mathur et al., 1996).  Ahmed et al. (2017); Mushtaq et al. (2016) and Pedhavi et al. (2017) also reported supplementation of ashwgandha led to decreased serum cholesterol. Reduced cholesterol levels in present study could be due to inhibitory effects of ashwagandha exerted at the levels of 3-hydroxyl-3-methyl-glutaryl-coA reductase, a key enzyme in cholesterol biosynthesis (Ansari et al., 2013; Ostlund, 2006) and increased excretion of cholesterol and bile acids through faecal sterol excretion (Ebihara and Schneeman, 1989). In the present study serum protein levels in all groups were not significantly influenced. On the contrary Sujatha et al. (2010) and Ansari et al. (2013) observed increased protein levels by feeding herbal mixture and ashwgandha respectively. Serum protein levels depends on availability of dietary protein, this suggests that proteins of ashwagandha fed diets were more available to the birds (Obikaonu et al., 2011.


The results of present study indicated that supplementation of Ashwagandha extract at 100mg/kg diet significantly decreased serum cholesterol levels without any deleterious effects on performance and did not show any significant effect on immunity.


  1. Ahmed, S. T. and Yang, C. J. 2017. Effects of Dietary Punica granatum By-products on Performance, Immunity, Intestinal and Fecal Microbiology, and Odorous Gas Emissions from Excreta in Broilers. The Journal of Poultry Science, 54(2): 157-166.
  2. Akotkar, N. S., Sarag, A. N., Rekhate, D. H., and Dhok, A. P. 2007. Effect of supplementation of Ashwagandha (Withania somnifera) on performance of broilers. Indian Journal of Poultry Science, 42(1): 92-94.
  3. Ansari, J., Khan, S. H., Haq, A. U., Ahmad, T., and Abbass, M. I. 2013. Effect of Supplementation of Withania somnifera (Linn.) Dunal Roots on Growth Performance, Serum Biochemistry, Blood Hematology, and Immunity of Broiler Chicks. Journal of herbs, spices & medicinal plants, 19(2): 144-158.
  4. Davis, L., and Kuttan, G. 2000. Immunomodulatory activity of Withania somnifera. Journal of Ethnopharmacology, 71(1): 193-200.
  5. Ebihara K. and B. O. Schneeman. 1989. Interaction of bile acids, phospholipids, cholesterol and triglyceride with dietary fibers in the small intestine of rats. J. Nutr.119: 1100-110.
  6. Edelman A S, Sachez P I, Robinson M E, Hochwalad G M and Thorbecke, G. J. 1986. Primary and Secondary Swelling response to Phytohaemagglutinin as a measure of immune competence in chickens. Avian Diseases, 30:105-111.
  7. Gambhir, D. S., Gambhir, J. K., and Sudha, R. (2000). Dyslipidemia and coronary heart disease: management issues from Indian perspective.
  8. Ghosal, S., Lal, J., Srivastava, R., Bhattacharya, S. K., Upadhyay, S. N., Jaiswal, A. K.and Chattopadhyay, U. 1989. Immunomodulatory and CNS effects of sitoindosides IX and X, two new glycowithanolides from Withania somnifera. Phytotherapy Research, 3(5): 201-206.
  9. Gross, W. B. 1992. Effect of short-term exposure of chickens to corticosterone on resistance to challenge exposure with Escherichia coli and antibody response to sheep erythrocytes. American journal of veterinary research, 53(3):291-293.
  10. Habibian, M., Ghazi, S., Moeini, M. M., and Abdolmohammadi, A. 2014. Effects of dietary selenium and vitamin E on immune response and biological blood parameters of broilers reared under thermoneutral or heat stress conditions. International journal of biometeorology, 58(5): 741-752.
  11. Hassan, A. M., Hussein, M. M., and Osman, M. M. 2007. Effect of chronic heat stress on broiler chicks performance and immune system. SCVMJ, 12: 55-68.
  12. Malik, F., Singh, J., Khajuria, A., Suri, K. A., Satti, N. K., Singh, S. and Qazi, G. N. 2007. A standardized root extract of Withania somnifera and its major constituent withanolide-A elicit humoral and cell-mediated immune responses by up regulation of Th1-dominant polarization in BALB/c mice. Life Sciences, 80(16): 1525-1538.
  13. Mashaly, M. M., Hendricks 3rd, G. L., Kalama, M. A., Gehad, A. E., Abbas, A. O., and Patterson, P. H. 2004. Effect of heat stress on production parameters and immune responses of commercial laying hens. Poultry Science, 83(6): 889-894.
  14. Mathur R, Sharma A, Dixit VP, Verma M 1996. Hypolipidaemic effect of fruit of officinalis in cholesterol fed rabbits. J. Ethnopharmacol, 50(2):61-68.
  15. Mujahid, A., Yoshiki, Y., Akiba, Y., and Toyomizu, M. 2005. Superoxide radical production in chicken skeletal muscle induced by acute heat stress. Poultry Science, 84(2): 307-314.
  16. Mushtaq, M., Durrani, F. R., Imtiaz, N., Sadique, U., Hafeez, A., Akhtar, S., and Ahmad, S. 2012. Effect of administration of Withania somnifera on some hematological and immunological profile of broiler chicks. Pak Vet J. 32: 70-72.
  17. Mushtaq, M., U. Sadique, N. Chand, I. Ahmed, S.S.A. Shah, I. Ahmad, Imran-Ullah and M. Shah. 2016. Biochemical effect of water based infusion of Withania somnifera in broiler production. Meat Sciences and Veterinary Public Health, 1(1):17-20.
  18. Nassir, F. Moundras, C. Bayle, D. Serougne, C. Gueux, E. Rock, E. and Mazur, A. 1997. Effect of selenium deficiency on hepatic lipid and lipoprotein metabolism in the rat. British Journal of Nutrition, 78(3): 493-500.
  19. Niu, Z. Y., Liu, F. Z., Yan, Q. L., and Li, W. C. 2009. Effects of different levels of vitamin E on growth performance and immune responses of broilers under heat stres s. Poultry science, 88(10): 2101-2107.
  20. Obikaonu, H. O., Okoli, I. C., Opara, M. N., Okoro, V. M. O., Ogbuewu, I. P., Etuk, E. B., and Udedibie, A. B. I. 2012. Haematological and serum biochemical indices of starter broilers fed leaf meal of neem (Azadirachta indica). Journal of Agricultural Technology, 8(1): 71-79.
  21. Ostlund, R. E., and Lin, X. 2006. Regulation of cholesterol absorption by phytosterols. Current atherosclerosis reports, 8(6): 487-491.
  22. Pedhavi, S., Burte, R. G., Kumar, S., Desai, B. G., Bhagat, D. J., Prasade, N. N., and Dhekale, J.S. 2017. Effect of Ashwagandha (Withania somnifera) on Haematology and Serum Biochemistry of Broiler Chicks.Advanced Agricultural Research & Technology Journal.
  23. Sahin N, Onderci M, Sahin K, Gursu MF, Smith MO 2004. Ascorbic acid and melatonin reduces heat induced performance inhibition and oxidative stress in Japanese quails. Br. Poult. Sci, 45: 116-122.
  24. Sahin, K., Sahin, N., and Yaralioglu, S. 2002. Effects of vitamin C and vitamin E on lipid peroxidation, blood serum metabolites, and mineral concentrations of laying hens reared at high ambient temperature. Biological trace element research, 85(1): 35-45.
  25. Singh, P. P., Sahu, D. S., Singh, M. K., Manoj, J., and Kumar, Y. 2016. Effect of supplementation of ashwagandha and vitamin E on haematological and immunological parameters of vencobb-400 broilers. Indian Journal of Poultry Science, 51(2): 164-167.
  26. Smith, M. O. 2003. Effects of different levels of zinc on the performance and immunocompetence of broilers under heat stress. Poultry science, 82(10): 1580-1588.
  27. Star, L., Decuypere, E., Parmentier, H. K., and Kemp, B. 2008. Effect of single or combined climatic and hygienic stress in four layer lines: 2. Endocrine and oxidative stress responses. Poultry science, 87(6): 1031-1038.
  28. Sujatha, V., Korde, J. P., Rastogi, S. K., Maini, S., Ravikanth, K., and Rekhe, D. S. 2010. Amelioration of heat stress induced disturbances of the antioxidant defense system in broilers. Journal of Veterinary Medicine and Animal Health, 2(3): 18-28.
  29. Uddin, Q., Samiulla, L., Singh, V. K. and Jamil, S. S. 2012. Phytochemical and Pharmacological Profile of Withania somnifera Dunal: A Review.
  30. Vasanthakumar, P., Pangayarselvi, B., Sasikumar, P., Chandrasekaran, D., Doraisamy, K. A., and Purushothaman, M. R. 2015. Performance of broilers fed ashwagandha (Withania somnifera) incorporated diets during summer season for alleviating heat stress. Indian Journal of Animal Research, 49(3): 333-335.
  31. Virden, W. S. and Kidd, M. T. (2009). Physiological stress in broilers: Ramifications on nutrient digestibility and responses. Journal of Applied Poultry Research18(2): 338-347.
  32. Wegmann, T. G., and Smithies, O. (1966). A simple hemagglutination system requiring small amounts of red cells and antibodies. Transfusion6(1): 67-73.
  33. Whitehead, C. C. and Keller, T. 2003. An update on ascorbic acid in poultry. World’s Poultry Science Journal, 59(2): 161-184.
Full Text Read : 3058 Downloads : 514
Previous Next

Open Access Policy