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Author(s): Sumit Kumar Dubey1, Krishan Kumar Verma2, Arifa Qureshi3, Neha Behar*4

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    1Department of Microbiology and Biotechnology, D.L.S. P.G. College, Bilaspur, Chhattisgarh, India
    2Department of Microbiology and Biotechnology, D.L.S. P.G. College, Bilaspur, Chhattisgarh, India
    3Department of Microbiology and Biotechnology, D.L.S. P.G. College, Bilaspur, Chhattisgarh, India
    4Department of Microbiology and Biotechnology, D.L.S. P.G. College, Bilaspur, Chhattisgarh, India
    *Corresponding Author Email:

Published In:   Volume - 4,      Issue - 2,     Year - 2022

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Sumit Kumar Dubey, Krishan Kumar Verma, Arifa Qureshi and Neha Behar (2022) Assessment of Antimicrobial Activity of Commercially available Withania Somnifera (Ashwagandha) Powder against Pathogenic microorganisms. A Review on various analytical methodology for Ondansetron. NewBioWorld A Journal of Alumni Association of Biotechnology,4(2):25-30.

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 NewBioWorld A Journal of Alumni Association of Biotechnology (2022) 4(2):25-30            


Assessment of Antimicrobial Activity of Commercially available Withania Somnifera (Ashwagandha) Powder against Pathogenic microorganisms

Sumit Kumar Dubey, Krishan Kumar Verma, Arifa Qureshi and Neha Behar*


Department of Microbiology and Biotechnology, D.L.S. P.G. College, Bilaspur, Chhattisgarh, India,

*Corresponding Author Email:




Article history:


25 September 2022

Received in revised form

28 October 2022


15 November 2022


Withania somnifer;

Indian ginseng;


human pathogens;

dietary supplement;

boost immunity


Withania somnifera L (or Ashwagandha or Indian ginseng) is a sub-tropical medicinal plant of Solanaceae family that is used by folk healers for multipurpose ailments in alternative system of treatment. It serves as bioactive ingredient in more than 100 ayurvedic formulations of Ayurveda, Siddha, and Unani medication. The ethnopharmacological application of Ashwagandha includes adaptogenic, anti-convulsion, anti-microbial anti-sedative, anti-stress, arthritis, geriatric debilities, mental disorders, and neurological disorders. The present study included validation of antimicrobial properties of commercially available W. Somnifera powder against human pathogens viz., P. aeruginosa (gram -ve), S. aureus (gram +ve), and E. coli (gram -ve). The experimental results exhibited that chloroform, isopropanol, and distilled water extract of W. somnifera showed significant antibacterial action, 15.5 mm, 20.5 mm, and 27 mm clear zones (in radius) against P. aeruginosa, S. aureus, and E. coli respectively. The prospects of Ashwagandha powder could be its applicability as a folklore pharmacological agent and by means of a harmless routine dietary supplement to boost immunity and prevent bacterial infection.



Withania somnifera is alternatively known as Ashwagandha or Indian ginseng or winter cherry and widely used as indigenous woody shrub around Asia and America. Indian Ayurveda used it as a medicinal herb since long ago to treat pain, inflammation, insomnia, and diabetes and, to increase reproductive efficacy, control bacterial infection and boost overall immunity. It has a broad spectrum of pharmacological potential towards inflammation, stress, tumor, neuroprotection, cardiac protection, maintaining cholesterol, and so forth. Moreover, it also helps in the regulation of apoptosis and stimulates endothelial function in the biological system. The Ashwagandha is used in herbal formulations e.g., ashwagandharishta, Ashwagandhadi Lehya, Balarishta, and alike since ancient times under the brand name Himalaya, Patanjali, Dabur, and Baidyanath. Indian Ayurveda practiced since 6000 BC. The Ashwagandha root is used as an effective Rasayana in Ayurveda. The Ashwagandha smells like ashwa (horse) hence it is called so. The antibacterial and antifungal action of Ashwagandha has been experimentally well-discussed in works of literature earlier (Kothalawala et al., 2019; Kandani et al., 2019; Gautam and Thakur, 2016; Sun et al., 2015; Dar et al., 2015; Soni et al., 2015; Bisht and Rawat 2014; Mwitari et al., 2013; Maurya, 2010; Sangwan et al., 2008; Gupta and Rana, 2007; Arora et al., 2004; Mishra et al., 2000;).

The initial phytochemical examination of W. somnifera showed that it comprises pharmacologically active alkaloids i.e., withanolides, Withanine, withanolide D, withaferin A (these account for around 38% of total alkaloids content) (Yoshida et al., 1979; Atal et al., 1975; Glotter et al., 1973; Lavie et al., 1972). Later, many literature authenticated presence of alkaloids, flavonoids, glycosides, phenolics, phytophenols, steroids, and saponins (Das et al., 2019; Divisha et al., 2018; Alam et al., 2011; Mirjalili et al., 2009).

DOI: 10.52228/NBW-JAAB.2022-4-2-5

Hence the present research was focused on the evaluation of the antibacterial efficacy of commercially available Ashwagandha powder against human pathogenic bacteria viz., Escherichia coli (resistant to Penicillin and Erythromycin), Pseudomonas aeuroginousa (resistant to Penicillin G and Aminopenicillin), Staphylococcus aureus (resistant to Penicillin and erythromycin). These pathogenic bacterial stains usually occupied the major fraction of nosocomial infection (Khan et al., 2015; Horan et al., 2008). The examination of commercially available Ashwagandha powder against human pathogenic bacteria could contribute towards the substitution of harmful antibiotics with Ashwagandha powder with the least or no side effects.


Materials and Methods

The commercially available Ashwagandha root powder of the Patanjali brand (an intact and undamaged pack) was purchased from an authorized Patanjali store located at Seepat Road, Bilaspur 495001 Chhattisgarh, India. The active cultures of test human pathogenic bacterial strains viz., E. coli (MTCC443), P. aeuroginosa (MTCC779), and S. aureus (MTCC737) were procured from the Department of Microbiology, D.L.S. PG. College, Bilaspur (C.G.) and stored in the refrigerator until used.

Sample preparation: The 2.0 g of Patanjali Ashwagandha root powder was taken in four sterile glass containers and 10 ml of solvent chloroform, acetone, distilled water, and isopropanol were poured into the glass container (one solvent in each glass container) mixed well using a clean glass rod to make a homogenous mixture and properly covered with silver foil (Figure 1). The aforementioned preparations were allowed to rest for 48 h. After the completion of the resting phase, the preparations were filtered using Whatman filter paper no.1 and collected. The filtered preparations were centrifuged (at 1000 rpm for 10 min) to remove reimaging insoluble particles. The clear supernatant was collected in clean air-tight glass and kept in a cool and dark place until used. These final preparations were further used to evaluate their antibacterial efficacy against test human pathogens.

Test pathogen culture preparation: Loopful culture from each active slant of E. coli, S. aureus, P. aeuroginosa were inoculated in 5.0 ml of broth culture separately and incubated overnight to prepare active inoculum under BSL2 guidelines as prescribed by the National Centre for Disease Control. 

Antimicrobial activity assay: The antimicrobial assay was examined using Disk Diffusion Method (DDM) also known as Kirby–Bauer test (CLSI, 2012). A 1.0 ml of active inoculums (matched to 0.5 McFarland standard, 1.5×108 CFU/ml) were spread over Muller Hilton Agar (MHA) plate and disk (diameter of 6.0 mm) flooded with Ashwagandha solvent extract and placed as per experimental design. The standard antibiotics (a broad-spectrum antibiotic erythromycin, 100 mg/ml) were used as control.

Result and Discussion

The present course of investigation was an attempt towards the validation of antimicrobial action of commercially available Withania Somnifera (Ashwagandha) Powder against human bacterial pathogens viz., P. aeruginosa (gram negative), S. aureus (gram positive), and E. coli (gram negative). The present result showed that chloroform, isopropanol, and distilled water extract of W. somnifera exhibited maximum antibacterial action of 15.5 mm, 20.5 mm, 27 mm clear zone (in radius) against P. aeruginosa, S. aureus, and E. coli respectively (Table 1 and Figure 2).

The present research investigation was carried out in commercial Ashwagandha powder to evaluate their antibacterial potential against human pathogens viz., E. coli (MTCC443), P. aeruoginosa (MTCC779), and S. aureus (MTCC737). E. coli causes nosocomial infections in the form of diarrhea, urinary tract infection (UTI), respiratory infection, and pneumonia while P. aeruginosa causes meningitis and septicemia malignant like nosocomial infections (Zafar et al., 2020; Jiménez-Mejías and Márquez-Rivas, 2020; Khan et al., 2015). Similarly, S. aureus also serves as a nosocomial pathogen in a hospital setting globally (Al-Talib et al., 2010).

Table 1: Antimicrobial profile of different solvent extracts of W. somnifera against test human pathogens

Solvent Extract of 

W. somnifera

The radius of clear zone (in mm) against test human pathogen

Pseudomonas aeruginosa

Staphylococcus aureus

Escherichia coli

Plate 1

Plate 2


Plate 1

Plate 2


Plate 1

Plate 2
















No zone

No zone





Distilled water




No zone

No zone









No zone

No zone






Arora et al. (2004) explored the antibacterial (Salmonella typhimurium and E. coli) properties of different extracts of Ashwagandha along with purified bioactive agents and revealed that the W. somnifera comprises of broad therapeutic value including ailments e.g., mental retardation, asthma, anti-inflammation, arthritis, tuberculosis, microbial infections, fever, and certain other diseases and disorders. Alam et al. (2012) assessed methanolic extracts of W. somnifera roots for antioxidant and antimicrobial activities and the result indicated significant antioxidant and antibacterial properties. Bisht and Rawat (2014) have been examined the leaf extract of W. sonmifera against methicillin-resistant S. aureus and observed that the methanolic W. somnifera leaf extract reflected maximum inhibition (20.6 mm) against test pathogen (S. aureus). Soni et al. (2015) investigated the antimicrobial potential of W. somnifera using the disk-diffusion method against bacterial pathogens e.g., S. aureus, Proteus vulgaris, P. aeuroginosa, and Bacillus subtilis. The maximum inhibition was observed with W. sonmifera ethanolic leaf extract. Further, numerous animal trials on Ashwagandha have been carried out. The Ashwagandha root has been reported as an effective remedy for Alzheimer’s by interfering with nuclear factor-kβ activation, inhibiting amyloid beta (Aβ) production, a revival of synaptic function, and improving antioxidant action via Nrf2 migration (Sandhir & sood 2017). Ashwagandha comprises of alkaloids, steroids, and saponins. The animal model (rat) based experiments revealed that the Ashwagandha boosts stamina in rats (evaluated by swimming endurance test) by altering ascorbic acid and cortisol content in the adrenal gland that is produced during swimming stress. The Ashwagandha has also been documented as an effective antitumor action against Chinese Hamster Ovary cell carcinoma and has been found a potent cognition-promoting action that is useful to treat children's and older aged person’s memory deficits (Alzoubi et al., 2019).

Both gram-positive and gram-negative bacterial strains are often associated with nosocomial clinical infections. The gram-negative can alter their outer membrane by changing hydrophobic properties or fostering mutations to increase antibiotic resistance that that of gram-positive bacteria. Our findings were most relevant to the previous work Singh G, Kumar P (2011), and Bisht and Rawat (2014). The present research work revealed that commercial Ashwagandha powder could potentially be used to mitigate both gram-positive and gram-negative bacterial infections.


Ashwagandha is a woody shrub known for its beneficial properties. The Ashwagandha powder is commercially available and widely used for health benefits due to its low-cost and sufficient availability. Ashwagandha has been extensively applicable for antimicrobial action and immunity boosters. The Ashwagandha chloroform, isopropanol, and distilled water extract exhibited maximum clear zone against P. aeruginosa, S. aureus, and E. coli respectively. The commercially available Ashwagandha powder was found to be effective against multidrug–resistant human pathogens. Hence it could potentially be utilized as an antibacterial agent and immunity booster.

Conflict of Interest

The authors declare that there is no conflict of interest.

Ethical Compliance Standard

There is no Ethical Compliance Standard.


The authors would be thankful to D.L.S. P.G. College, Bilaspur for the well-equipped research facility.


Alam N, Hossain M, Khalil MI, Moniruzzaman M, Sulaiman SA, Gan SH (2011) High catechin concentrations detected in Withania somnifera (ashwagandha) by high performance liquid chromatography analysis. Altr Med., 11:65–69.

Alam N, Hossain M, Mottalib MA, Sulaiman SA, Gan, SH, Khalil MI (2012) Methanolic extracts of Withania somnifera leaves, fruits and roots possess antioxidant properties and antibacterial activities. BMC Complementary and Alternative Medicine, 12(1).

Al-Talib HI, Yean CY, Al-Jashamy K, Hasan H (2010) Methicillin-resistant Staphylococcus aureus nosocomial infection trends in Hospital University Sains Malaysia during 2002-2007. Ann Saudi Med., 30(5):358-63.

Alzoubi KH, Al Hilo AS, Al-Balas QA, El-Salem K, El-Elimat T, Alali FQ (2019) Withania somnifera root powder protects against post-traumatic stress disorder-induced memory impairment. Mol Biol Rep., 46(5):4709-4715.   

Arora S, Dhillon S, Rani G, Nagpal A (2004) The in vitro antibacterial/synergistic activities of Withania somnifera extracts. Fitoterapia, 75(3-4):385-8.

Atal CK, Dhar KL, Gupta OP, Raghunathan K (1975) Pharmacognosy and phytochemistry of Withania somnifera (Linn ) Dunal (Ashwagandha) New Delhi: Central Council for Research in Indian Medicine and Homeopathy.

Bisht P, Rawat V (2014) Antibacterial activity of Withania somnifera against Gram-positive isolates from pus samples. Ayu. 35(3):330-2.

CLSI (2012) Performance Standards for Antimicrobial Disk Susceptibility Tests, Approved Standard, 7th ed., CLSI document M02-A11. Clinical and Laboratory Standards Institute, Pennsylvania 19087, USA.

Dar NJ, Hamid A, Ahmad M (2015) Pharmacologic overview of Withania sonmifera, The Indian Ginseng. Cell Mol. Life Sci., 72(2015): 4445-4460.

Das S, Saraf A, Sharma D, Sohal JK (2019) Qualitative screnning of bioactive secondary metabolites present in Withania somnifera and Rauwolfia serpentina root and stem extract with pharmacological importance. Int J Res Ana Rev., 6:69–74.

Divisha R, Ranganathan V, Vijayakaran K, Elamaran A, Senthil KP (2018) Quantifying phytophenols in Andrographis paniculata and Withania somnifera leaf extract. J Pharam., 7:477–479.

Gautam A, Thakur MK (2016) Assessment of cholinergic properties of Ashwagandha leaf - extract in the amnesic mouse brain. Ann neurosci, 23: 68-75.

Glotter E, Kirson I, Abraham A, Lavie D (1973) Constituents of Withania somnifera (Dunal) XIII—the withanolides of chemotype III. Tetrahed., 29:1353–1364.

Gupta GL, Rana AC (2007) Plant review Withania somnifera (Ashwagandha). Pharmacognosy reviews, 1(1).

Horan TC, Andrus M, Dudeck MA (2008) CDC/NHSN surveillance definition of health care–associated infection and criteria for specific types of infections in the acute care setting. American Journal of Infection Control, 36(5):309–332.

Jiménez-Mejías ME, Márquez-Rivas FJ (2020) CNS infections in patients with temporary CSF shunts: Diagnostic, preventive and therapeutic approach. In Enfermedades Infecciosas. Microbiología Clínica, 38(2): 49–53.

Kandani N, Sahu P, Kalwani T, Ardeshna A, Dharajiya D (2019) Antibacterial and antifungal activities of ashwagandha (Withania somnifera). Journal of Durg Delivery and Therapeutics, 9(5-5):154-161.

Khan HA, Ahmad A, Mehboob R (2015) Nosocomial infections and their control strategies. Asian Pacific Journal of Tropical Biomedicine, 5(7): 509–514.

Kothalawala M, Jayaweera JAAS, Arunan S (2019) The emergence of non-albicans candidemic and evaluation of Hi Chrome candida differential agar and VITEK2 YST® Platform for differential of candida bloodstream isolates in teaching hospital Kandy, Sri lanka. BMC Microbiol, 19:136.

Lavie D, Kirson I, Glotter E, Rabinovich D, Shakked Z (1972) Crystal and molecular structure of withanolide E, a new natural steroidal lactone with a 17α–side–chain. J Chem Soc Chem Comm., 15:877–878.

Maurya R (2010) Chemistry and pharmacology of Withania sonmifera coagulans: an Ayurvedic remedy Taxonomical classification phytochemistry pharmacological properties. Journal of pharmacy and pharmacology, 62:153-160.

Mirjalili MH, Moyano E, Bonfill M, Cusido RM, Palazon J (2009) Steroidal lactones from Withania somnifera, an ancient plant for novel medicine. Mole., 14: 2373–2393.

Mishra LC, Singh BB, Dagenais S (2000) Scientific basis for the therapeutic use of Withania sonmifera, 5(4).

Mwitari PG, Ayeka PA, Ondicho J, Matu EN, Bii CC (2013) Antimicrobial Activity and Probable Mechanisms of Action of Medicinal Plants of Kenya: Withania somnifera, Warbugia ugandensis, Prunus africana and Plectrunthus barbatus. PLoS one, 8(6):e65619.

Sangwan RS, Das CN, Lal P, Misra L, Tuli R, Sangwan NS (2008) Withanolide A is inherently de novo biosynthesized in roots of the medicinal plant Ashwagandha (Withania somnifera). Physiol Plant., 133(2):278-87.

Singh G, Kumar P (2011) Evaluation of antimicrobial efficacy of flavonoids of Withania somnifera L. Indian J Pharm Sci., 73(4):473-8.

Soni P, Bahadur AN, Tewari U (2015) Study on antimicrobial activity of leaf extract of Withania somnifera L. Dunal against clinical pathogens. International Journal of Advanced Research in Biological Sciences, 2(11):193–196.

Sun C, Wu Z, Wang Z, Zhang H (2015) Effect of Ethanol/Water Solvents on Phenolic Profiles and Antioxidant Properties of Beijing Propolis Extracts. Evidence-Based Complementary and Alternative Medicine, 2015: 1–9.

Yoshida M, Hoshi A, Kuretani K, Ishiguro M, Ikekawa N (1979) Relationship between chemical structure and antitumor activity of withaferin A analogues. J Pharmacobiodyn., 2:92–97.

Zafar M, Tauseef A, Asghar MS, Khan N, Farooqui N, Dawood M, Alam T, Naman D (2020) Escherichia coli: a rare cause of meningitis in immuno-competent adult. J Community Hosp Intern Med Perspect. 10(1):69-72.




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