WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Prashith et al. World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 2.786

Volume 3, Issue 8, 1483-1488. Research Article ISSN 2278 – 4357

ANTIMICROBIAL ACTIVITY OF GRAPE SEED EXTRACT

Ranjitha C.Y, Priyanka S, Deepika R, Smitha Rani G.P, Sahana J,

Prashith Kekuda T.R*

Department of Microbiology, S.R.N.M.N College of Applied Sciences, N.E.S Campus,

Balraj Urs Road, Shivamogga-577201, Karnataka, India

Article Received on ABSTRACT

11 May 2014, Grape pomace (skin and seed) is produced during winemaking and is
Revised on 14 June
2014, considered to be a waste product of wine industries. Grape seed is
Accepted on 26 July 2014
known to exhibit various bioactivities. In the present study, we
determined antimicrobial activity of grape seed extract (GSE). Agar
*Correspondence for Author
well diffusion assay was conducted to determine inhibitory effect of
Prashith Kekuda T.R
Department of Microbiology, GSE against Staphylococcus aureus, Escherichia coli and Klebsiella
S.R.N.M.N College of Applied pneumoniae isolated from urinary tract infection. S. aureus and E. coli
Sciences, N.E.S Campus, was inhibited to high and least extent respectively by GSE. Poisoned
Balraj Urs Road, Shivamogga-
food technique was performed to screen antifungal effect of GSE
577201, Karnataka, India
against Colletotrichum capsici. GSE was effective against mycelia
growth of C. capsici (>65% inhibition). GSE can be used in the treatment of urinary tract
infections and in the management of anthracnose of chilli.

Key words: Grape seed extract, Agar well diffusion, Urinary tract infection, Poisoned food
technique, Colletotrichum capsici.

INTRODUCTION
Grape (Vitis vinifera L.) is one among the important and leading fruit crops grown
worldwide. About 80% of the harvest is used in industries for wine making. The solid grape
residues (pomace) namely skin and seeds are produced in large quantities by the winemaking
industries. These are typically waste byproducts of wine industries. Grape seeds are around
15% of the solid waste produced in wine industries. Grape seed is a well-known dietary
supplement and contains vitamins, minerals, and polyphenols. The abundant phenolic
compounds from grape seed are catechins, epicatechin, procyanidin, and some dimmers and
trimers. The polyphenols of grape seeds have been recognized for their beneficial role in

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human health. The grape seed is shown to exhibit bioactivities such as antioxidant, anti-
inflammatory, anti-bacterial, anti-cancer, antiviral, cardioprotective, hepatoprotective,
neuroprotective, antiaging and anti-diabetic. The oil extracted from grape seeds is used in
cosmetic, culinary, pharmaceutical and medical purposes[1-6]. In the present study, we
determined antimicrobial activity of grape seed extract (GSE) against urinary tract bacteria
and Colletotrichum capsici.

MATERIALS AND METHODS

Preparation of GSE
Fully ripe grapes were purchased from a local shop in market of Shivamogga city. The grapes
were crushed and seeds were separated. The seeds were washed well using clean water and
dried in oven at 60oC. The seeds were powdered in a blender. 25g of grape seed powder was
added to a conical flask containing 100ml of methanol and stirred well. The flask was left
aside for 48 hours and occasionally stirred. The content of flask was filtered through
Whatman No. 1 and evaporated to dryness in oven a 50oC[7].

Antibacterial activity of GSE

Agar well diffusion assay was performed to investigate antibacterial efficacy of GSE against
three bacteria viz., Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus from
urinary tract infections. 24 hours old Nutrient broth (HiMedia, Mumbai) cultures of test
bacteria were swabbed uniformly on sterile Nutrient agar (HiMedia, Mumbai) plates. Using
sterile cork borer, wells of 8mm diameter were punched in the inoculated plates. 100µl of
GSE (20mg/ml of 25% Dimethyl sulfoxide [DMSO; HiMedia, Mumbai]), reference antibiotic
(Chloramphenicol, 1mg/ml of sterile distilled water) and DMSO (25%, in sterile water) were
added to labeled wells and the plates were incubated for 24 hours at 37oC. The zones of
inhibition around the wells were measured using a ruler[7].

Antifungal activity of GSE

In order to determine antifungal effect of GSE against C. capsici, we employed poisoned
food technique with minor modifications[8]. The spore suspension of test fungus was
inoculated at the centre of control plates (without extract) and poisoned Potato dextrose agar
(HiMedia, Mumbai) plates (1mg extract/ml of medium) by point inoculation method using
sterile inoculation needle. The plates were incubated for 5 days. The diameter of fungal
colonies on control and poisoned plates was measured on 5th day. Antifungal activity of GSE
was mentioned as

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Prashith et al. World Journal of Pharmacy and Pharmaceutical Sciences

% Inhibition of mycelial growth = (C - T / C) x100, where C is the diameter of colonies on

control plates and T is the diameter of colonies on poisoned plates.

RESULTS
Antibacterial activity of GSE
The result of antibacterial efficacy of GSE is shown in Table 1. GSE was found to inhibit all
test bacteria but to a varied extent. Among bacteria, marked inhibition was observed in case
of S. aureus whereas E. coli was inhibited to least extent. Inhibitory activity of reference
antibiotic was higher than that of GSE. DMSO was not effective in inhibiting test bacteria.

Table 1: Inhibitory activity of GSE against test bacteria

Zone of inhibition in cm
Test bacteria
GSE Antibiotic DMSO
E. coli 1.1±0.0 3.1±0.2 0.0±0.0
K. pneumoniae 1.3±0.1 2.2±0.0 0.0±0.0
S. aureus 1.8±0.0 3.3±0.2 0.0±0.0
Antifungal activity of GSE
The GSE was found to inhibit mycelial growth of C. capsici and the result is shown in Table
2. The growth of C. capsici was drastically reduced on plates poisoned with GSE (with an
inhibition of 68%) when compared to control plates.

Table 2: Inhibitory activity of GSE against C. capsici

Treatment Colony diameter (cm) Inhibition (%)
Control 3.2±0.1 -
GSE 1.0±0.0 68.75
DISCUSSION
Urinary tract infections (UTIs) are one among the most common infections in community and
hospital settings affecting individuals of all age groups. Several bacteria are known to cause
UTIs among which E. coli is involved in most cases. The infections may involve single
species or sometimes it may be polymicrobial in nature. UTIs are treated using antibiotics.
However, most strains have already developed resistance against commonly used antibiotics.
Hence, research on alternative disease treatment strategies is of much interest. It has been
already shown that plants exhibit marked inhibitory efficacy against uropathogenic
bacteria[9-16]. In the present study, we screened the efficacy of GSE against three bacterial
isolates recovered from urinary tract infections. The GSE was more effective against S.
aureus followed by K. pneumoniae and E. coli i.e., susceptibility to GSE was high in case of
Gram positive bacterium when compared to Gram negative bacteria. Similar result was

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Prashith et al. World Journal of Pharmacy and Pharmaceutical Sciences

observed in the study of Jayaprakasha et al.[17] where solvent extracts of grape seed powder
inhibited Gram positive bacteria at lower concentration while Gram negative bacteria were
inhibited at high concentration. In another study, Corrales et al.[18] showed the efficacy of
GSE against food borne pathogens. It was observed that Gram positive pathogens were
inhibited by GSE while Gram negative pathogens remained unaffected.

The term chilli refers to ripe fruit of the genus Capsicum (Solanaceae). Chilli is extensively
grown for consumption, nutritional and economy purposes. Anthracnose is an important
disease of chilli (Capsicum annuum L.). The disease results in drastic loss in yield and
deterioration of fruit quality. The typical symptoms of anthracnose include sunken necrotic
tissues with concentric rings of acervuli on chilli fruit. The disease is caused by various
species of genus Colletotrichum among which C. capsici is an important pathogen. The
management of disease is mainly based on the use of synthetic fungicides. However, the
pathogen is reported to exhibit resistance against synthetic fungicides. Hence, search for
cheaper, ecofriendly alternative strategies for disease control is of interest. Plants are one of
the best alternatives for management of fungal diseases[8,19-21]. In the present study, we
evaluated antifungal efficacy of GSE against C. capsici isolated from anthracnose of chilli.
GSE was found to exhibit marked inhibition of mycelial growth of fungus. An inhibition of
>65% of fungus by extract was observed in this study. In an earlier study, Mendoza et al.[22]
showed inhibitory effect of anthocyanin rich extracts from grape pomace against
phytopathogenic fungus Botrytis cinerea.

CONCLUSION
Grape pomaces are the major byproduct, often considered as waste, generated during
winemaking process. The utilization of grape seeds is advantageous in terms of waste
reduction and production of added value products. In the present study, the GSE was found to
exhibit marked antibacterial and antifungal activity. The GSE can be used to treat urinary
tract infections and to control anthracnose disease in chilli.

ACKNOWLEDGEMENTS
Authors are thankful to Head, Department of Microbiology, Principal, S.R.N.M.N College of
Applied Sciences, Shivamogga and N.E.S, Shivamogga for providing facilities to conduct
work.

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