Journal of Alumni Association of Biotechnology (2022) 4(2):25-30
Antimicrobial Activity of Commercially available Withania Somnifera (Ashwagandha) Powder against Pathogenic
Sumit Kumar Dubey, Krishan Kumar Verma, Arifa Qureshi and
Department of Microbiology and
Biotechnology, D.L.S. P.G. College, Bilaspur, Chhattisgarh, India
*Corresponding Author Email: firstname.lastname@example.org
25 September 2022
Received in revised form
28 October 2022
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
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).
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
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
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
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).
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
Table 1: Antimicrobial
profile of different solvent extracts of W.
somnifera against test human pathogens
Solvent Extract of
The radius of clear zone (in mm)
against test human pathogen
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.,
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
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
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.
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