Effect of Ashwagandha (Withania somnifera) root extract enriched shrikhand on Salmonella infected albino mice: Effect of Ashwagandha enriched shrikhand on albino mice

Ashok Kumar Yadav; Dinesh Chandra Rai; Aman Rathaur

doi:10.62310/liab.v5i2.231

Authors

Ashok Kumar Yadav Department of Food Technology, Rajiv Gandhi University, Rono Hills, Doimukh, Itanagar, India https://orcid.org/0000-0002-7863-6143
Dinesh Chandra Rai Department of Dairy Science and Food Technology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi, India
Aman Rathaur School of Advanced Agriculture Sciences and Technology, Chhatrapati Shahu Ji Maharaj University, Kanpur, India https://orcid.org/0000-0003-3000-3301

DOI:

https://doi.org/10.62310/liab.v5i2.231

Keywords:

Ashwagandha root, Serum lipids, Growth performance, Immunomodulatory activity, Mice

Abstract

Herbs have long been utilized as food flavor, preservatives, and medicinal ingredients. The current study investigated the effects of Ashwagandha (Withania somnifera (L.) root extract enriched shrikhand on growth performance, blood lipid profile, liver enzyme activities, bacterial counts, serum immunoglobulin, and spleen lymphocyte activity in albino mice infected with Salmonella typhi. A total of 72 albino mice (20-25 days) were fed a basal diet for one week, followed by Salmonella infection. After one week, the mice were randomly distributed into three treatment groups for 23 days: T1 (basal diet), T2 (basal diet + 0.7 g/kg shrikhand), and T3 (basal diet + 0.7 g/ kg Ashwagandha root extract enriched shrikhand). Compared with the other treatment groups, the T3 group displayed the highest body weight, with no significant difference in feed intake across the treatments. However, compared to T1 group, the blood lipid profiles in the T2 and T3 groups significantly improved. Lipid peroxidation was significantly (p<0.05) reduced in mice fed Ashwagandha root extract-enriched shrikhand accompanied by increased superoxide dismutase and catalase activities. Salmonella counts in the small intestine and fecal matter were significantly (p<0.05) lower in the T3 group than in T1 and T2 groups. Serum immunoglobulin and spleen lymphocytes and immunoglobulin production was significantly enhanced in T3 group compared to others. According to the results of the present study, the Ashwagandha root extract-enriched shrikhand has immunomodulatory, antioxidant, antibacterial, and lipid-lowering effects in Salmonella-infected mice, highlighting its potential application as a functional food for immune enhancement and infection management.

Metrics

Metrics Loading ...

References

Attia G, El-Eraky W, Hassanein E, El-Gamal M, Farahat M, Hernandez-Santana A. (2017). Effect of dietary inclusion of a plant extract blend on broiler growth performance, nutrient digestibility, caecal microflora and intestinal histomorphology. International Journal of Poultry Science 16(9): 344-353. https://doi.org/10.3923/ijps.2017.344.353

Ayoub M, Lallouette P, Sütterlin BW, Bessler WG, Huber M, Mittenbühler K. (2003). Modulation of the Th1/Th2 bias by an immunoglobulin histamine complex in the ovalbumin allergy mouse model. International Immunopharmacology 3(4): 523-539. https://doi.org/10.1016/S1567-5769(03)00031-6

Azimi V, Mirakzehi MT, Saleh H. (2020). Hydroalcoholic extract of Withania somnifera leaf and α-tocopherol acetate in diets containing oxidised oil: effects on growth performance, immune response, and oxidative status in broiler chickens. Italian Journal of Animal Science 19(1): 917-928. https://doi.org/10.1080/1828051X.2020.1808537

Birla H, Keswani C, Rai SN, Singh SS, Zahra W, Dilnashin H, Rathore AS, Singh SP. (2019). Neuroprotective effects of Withania somnifera in BPA induced-cognitive dysfunction and oxidative stress in mice. Behavioral and Brain Functions 15: 9. https://doi.org/10.1186/s12993-019-0160-4

Chandrasekaran S, Veronica J, Sundar S, Maurya R. (2017). Alcoholic fractions F5 and F6 from Withania somnifera leaves show a potent Antileishmanial and Immunomodulatory activities to control experimental Visceral Leishmaniasis. Frontiers in medicine 4: 55. https://doi.org/10.3389/fmed.2017.00055

Chandrasekhar K, Kapoor J, Anishetty S. (2012). A prospective, randomized double-blind, placebo-controlled study of safety and efficacy of a high-concentration full-spectrum extract of Ashwagandha root in reducing stress and anxiety in adults. Indian Journal of Psychological Medicine 34: 255-262. https://doi.org/10.4103/0253-7176.106022

Ehuwa O, Jaiswal AK, Jaiswal S. (2021). Salmonella, food safety and food handling practices. Foods 10:9 07. https://doi.org/10.3390/foods10050907.

Elsemelawy SAE, Tag Al Deen R. (2016). Effect of feeding cake fortified with different concentrations of Ashwagandha root against sodium arsenite toxicity in male rats. Bulletin of the National Nutrition Institute of the Arab Republic of Egypt 47(2): 1-22. https://doi.org/10.21608/bnni.2016.4215

Felix J, Kanetkar P, Bunkar DS, Rose H, Paswan VK. (2025). Study on Quality Characteristics and Nutraceutical Properties of Paneer Fortified with Ashwagandha (Withania somnifera) Powder. Asian Journal of Dairy and Food Research. DR-2294: 1-9. https://doi.org/10.18805/ajdfr.DR-2294

Fujii J, Homma T, Osaki T. (2022). Superoxide radicals in the execution of cell death. Antioxidants 11: 501. https://doi.org/10.3390/antiox11030501

Grand View Research (2019). Functional foods market size, share & trends analysis report by ingredient (carotenoids, prebiotics & probiotics, fatty acids, dietary fibers), by product, by application, by region, and segment forecasts, 2022-2030. https://www.grandviewresearch.com/industry-analysis/functional-food-market, Accessed 12 May 2025.

Hamelet J, Demuth K, Dairou J, Ledru A, Paul JL, Dupret JM, Janel N. (2007). Effects of catechin on homocysteine metabolism in hyperhomocysteinemic mice. Biochemical and Biophysical Research Communications 355(1): 221-227. https://doi.org/10.1016/j.bbrc.2007.01.142

He JW, Zhou XJ, Lv JC, Zhang H. (2020). Perspectives on how mucosal immune responses, infections and gut microbiome shape IgA nephropathy and future therapies. Theranostics 10(25): 11462-11478. https://doi.org/10.7150/thno.49778

Ismaiel A, Dumitraşcu DL. (2019). Cardiovascular risk in fatty liver disease: the liver-heart axis—literature review. Frontiers in Medicine 6: 202. https://doi.org/10.3389/fmed.2019.00202

Joshi VK. Joshi A. (2021). Rational use of Ashwagandha in Ayurveda (Traditional Indian Medicine) for health and healing. Journal of Ethnopharmacology 276: 114101. https://doi.org/10.1016/j.jep.2021.114101

Jyotsana PK, Berwal R. (2019). Supplementation of Ashwagandha root powder improving gut micro flora and immune response of broilers. The Pharma Innovation Journal 8: 114-118.

Khabiya R, Choudhary GP, Jnanesha AC, Kumar A, Lal RK. (2024). An insight into the potential varieties of Ashwagandha (Indian ginseng) for better therapeutic efficacy. Ecological Frontiers 44(3): 444-450. https://doi.org/10.1016/j.chnaes.2023.06.009

Khan S, Malik F, Suri KA, Singh J. (2009). Molecular insight into the immune up-regulatory properties of the leaf extract of Ashwagandha and identification of Th1 immunostimulatory chemical entity. Vaccine 27: 6080-6087. https://doi.org/10.1016/j.vaccine.2009.07.011

Kumar V, Verma T, Chandra S, Wanjari A, Patel V, Singh S. (2024). Study of Phytochemical Attributes of Ashwagandha Ghrita from Desi Cow Milk. Asian Journal of Dairy and Food Research 43(3): 453-461. https://doi.org/10.18805/ajdfr.DR-2229

Kuo-Haller P, Cu Y, Blum J, Appleton JA, Saltzman WM (2010). Vaccine delivery by polymeric vehicles in the mouse reproductive tract induces sustained local and systemic immunity. Molecular pharmaceutics, 7(5): 1585-1595. https://doi.org/10.1021/mp100009e

Langade D, Dawane J, Dhande P. (2023). Sub-acute toxicity of Ashwagandha (Withania somnifera) root extract in wistar rats. Toxicology Reports 11: 389-395. https://doi.org/10.1016/j.toxrep.2023.10.009

Lin T, Brunner T, Tietz B, Madsen J, Bonfoco E, Reaves M, Huflejt M, Green DR. (1998). Fas ligand- mediated killing by intestinal intraepithelial lymphocytes. Participation in intestinal graft-versus-host disease. Journal of Clinical Investigation 101(3): 570-577. https://doi.org/10.1172/JCI896

Lorenz TK, Demas GE, Heiman JR. (2015). Interaction of menstrual cycle phase and sexual activity predicts mucosal and systemic humoral immunity in healthy women. Physiology and Behavior 152: 92-98. https://doi.org/10.1016/j.physbeh.2015.09.018

Maji S, Bumbadiya M, Sao K. (2023). Value addition of traditional indian dairy products using herbs and spices-An overview. International Journal of Bio-resource and Stress Management 14(3): 400-406. https://doi.org/10.23910/1.2023.3363a

Malik F, Singh J, Khajuria A, Suri KA, Satti NK, Singh S, Qazi GN. (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. https://doi.org/10.1016/j.lfs.2007.01.029

Mandlik DS, Namdeo AG (2021). Pharmacological evaluation of Ashwagandha highlighting its healthcare claims, safety, and toxicity aspects. Journal of Dietary Supplements 18(2): 183-226. https://doi.org/10.1080/19390211.2020.1741484

Mathur R, Oh H, Zhang D, Park SG, Seo J, Koblansky A, Ghosh S. (2012). A mouse model of Salmonella typhi infection. Cell 151:590-602. https://doi.org/10.1016/j.cell.2012.08.042

Mikulska P, Malinowska M, Ignacyk M, Szustowski P, Nowak J, Pesta K, Cielecka-Piontek J. (2023). Ashwagandha (Withania somnifera)-Current research on the health-promoting activities: a narrative review. Pharmaceutics 15:1057. https://doi.org/10.3390/pharmaceutics15041057

Mirakzehi MT, Hosseini SJ, Saleh H. (2017). The effects of hydroalcoholic extracts of Withania somnifera root, Withania coagulans fruit and 1,25-dihydroxycholecalciferol on immune response and small intestinal morphology of broiler chickens. Journal of Applied Animal Research 45(1): 591-597. https://doi.org/10.1080/09712119.2016.1236725

Mirakzehi MT, Hosseini SJ, Saleh H. (2018). Effects of two plant extracts and vitamin D3 on bone mechanical and histological properties of broiler chickens. Journal of Animal Plant Science 28(3): 686-694.

Mohammadi M, Mirakzehi MT, Saleh H, Baranzehi T. (2022). The effects of hydroalcoholic extract of Withania Somnifera leaf and fish oil on growth performance, bone calcification, morphological and mechanical characteristics and gene expression in broiler chickens. Poultry Science Journal 10: 169-183. https://doi.org/10.22069/psj.2022.19864.1768

Mondal P, Halder P, Mallick B, Bhaumik S, Koley H, Dutta S, Dutta M. (2023). Controlling the bacterial load of Salmonella Typhi in an experimental mouse model by a lytic Salmonella phage STWB21: a phage therapy approach. BMC Microbiology 23: 324. https://doi.org/10.1186/s12866-023-03040-3

Mossa ATH, Heikal TM, Belaiba M, Raoelison EG, Ferhout H, Bouajila J. (2015). Antioxidant activity and hepatoprotective potential of Cedrelopsis grevei on cypermethrin induced oxidative stress and liver damage in male mice. BMC Complementary And Alternative Medicine 15: 251. https://doi.org/10.1186/s12906-015-0740-2

Naushad S, Ogunremi D, Huang H. (2023). Salmonella: A brief review. In: Huang H, Naushad S, editors, Salmonella - Perspectives for low-cost prevention, control, and treatment. Intech Open, London. Pp 1-23. https://doi.org/10.5772/intechopen.112948

Naveed MA, Kim J, Ansari MA, Kim I, Massoud Y, Kim J, Rho J. (2022). Single-step fabricable flexible metadisplays for sensitive chemical/biomedical packaging security and beyond. ACS Applied Materials and Interfaces 14(27): 31194-31202. https://doi.org/10.1021/acsami.2c09628

Nijnik A, Dawson S, Crockford TL, Woodbine L, Visetnoi S, Bennett S, Cornall RJ. (2009). Impaired lymphocyte development and antibody class switching and increased malignancy in a murine model of DNA ligase IV syndrome. Journal of Clinical Investigation 119(6): 1696-1705. https://doi.org/10.1172/JCI32743

NRC (1995). Subcommittee on laboratory animal nutrition, Nutrient requirements of laboratory animals, Fourth Revised Edition, National Academies Press (US), Washington (DC).

Nurjayadi M, Kartika IR, Kurniadewi F, Nurasiah N, Ariastuti D, Sulfianti A, Wardoyo, WM. (2018). Immunogenicity evaluation of recombinant fim-c Salmonella typhi protein as typhoid vaccine candidate on wistar rat. IOP Conference Series: Materials Science and Engineering 434: 012094. https://doi.org/10.1088/1757-899X/434/1/012094

Parasuraman S, Raveendran R, Kesavan R. (2010). Blood sample collection in small laboratory animals. Journal of Pharmacology and Pharmacotherapeutics 1(2): 87-93. https://doi.org/10.4103/0976-500X.72350

Patel S, Thakur V, Parmar H. (2019). Effect of Withania somnifera (Ashwagandha) leaves extract on L-Methionine induced hyperhomocysteinemia in rats. International Journal of Pharmacognosy 6: 75-79. http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.6(2).75-79

Paul S, Chakraborty S, Anand U, Dey S, Nandy S, Ghorai M, Dey A. (2021). Withania somnifera (L.) Dunal (Ashwagandha): A comprehensive review on ethnopharmacology, pharmacotherapeutics, biomedicinal and toxicological aspects. Biomedicine & Pharmacotherapy 143: 112175. https://doi.org/10.1016/j.biopha.2021.112175

Rizwana H, Hazzani AAA, Shehata AI, Moubayed NMS. (2012). Antibacterial potential of Withania somnifera L. against human pathogenic bacteria. African Journal of Microbiology Research 6: 4810-4815. https://doi.org/10.5897/ajmr12.660

Rotimi SO, Ojo DA, Talabi OA, Balogun EA, Ademuyiwa O. (2012). Tissue dyslipidemia in Salmonella-infected rats treated with amoxillin and pefloxacin. Lipids in Health and Disease 11: 1-11. https://doi.org/10.1186/1476-511X-11-152

Savaliya V, Ahuja KK, Thesiya AJ, Hazra T. (2023). Development and evaluation of ginger-honey Shrikhand–a fermented sweet delicacy. Indian Journal of Dairy Science 76(4): 317-325. https://doi.org/10.33785/IJDS.2023.v76i04.002

Sethi S, Tyagi SK, Anurag RK. (2016). Plant-based milk alternatives an emerging segment of functional beverages: a review. Journal of Food Science and Technology 53: 3408-3423. https://doi.org/10.1007/s13197-016-2328-3

Shamji MH, Valenta R, Jardetzky T, Verhasselt V, Durham SR, Würtzen PA, van Neerven RJ. (2021). The role of allergen‐specific IgE, IgG, and IgA in allergic disease. Allergy 76: 3627-3641. https://doi.org/10.1111/all.14908

Shao T, Hsu R, Rafizadeh DL, Wang L, Bowlus CL, Kumar N, Leung PS. (2023). The gut ecosystem and immune tolerance. Journal of Autoimmunity 141: 103114. https://doi.org/10.1016/j.jaut.2023.103114

Sheikh R, Gallehdari M. (2023). The effect of herbal supplement and exercise training on plasma lipid profile in diabetic male rats. Journal of Exercise & Organ Cross Talk 3(2): 86-92. https://doi.org/10.22034/jeoct.2023.399703.1083

Shimmi SC, Jahan N, Sultana N. (2012). Effects of Ashwagandha (Withania somnifera) root extract against gentamicin induced changes of serum electrolytes in rats. Journal of Bangladesh Society of Physiologist 7(1): 29-35. https://doi.org/10.3329/jbsp.v7i1.11157

Spiteller G. (2003). Are lipid peroxidation processes induced by changes in the cell wall structure and how are these processes connected with diseases? Medical Hypotheses 60: 69-83. https://doi.org/10.1016/S0306-9877(02)00333-X

Yadav AK, Rai DC, Rathaur A. (2024). Process optimization for the development of Ashwagandha root extract enriched shrikhand using response surface methodology. Journal of Food Chemistry and Nanotechnology 10(S1): S47-S56. https://doi.org/10.17756/jfcn.2024-s1-007

Yadav AK, Rai DC. (2018). In vitro screening of Ashwagandha root extracts for the maximum functional components. The Pharma Innovation Journal 7(2): 12-16.

Yamada K, Hung P, Park TK, Park PJ, Lim BO. (2011). A comparison of the immunostimulatory effects of the medicinal herbs Echinacea, Ashwagandha, and Brahmi. Journal of Ethnopharmacology 137(1): 231-235. https://doi.org/10.1016/j.jep.2011.05.017

Yan L, Combs GFJr, DeMars LC, Johnson LK. (2011). Effects of the physical form of the diet on food intake, growth, and body composition changes in mice. Journal of the American Association for Laboratory Animal Science 50(4): 488-494.