eJBio Electronic Journal of Biology, 2009, Vol.

5(2): 40-44

Antibacterial Effect of Herbs and Spices Extract on

Escherichia coli
Venugopal Amrita, Dasani Sonal*, Rai Shalini
Department of Life Sciences, Kishinchand Chellaram College, D.W Road, Churchgate, Mumbai-
400 020 India.

* Corresponding author. Tel: +919820291092; E-mail: sonaldasani@[Link]

can cause gastroenteritis, urinary tract infections,

Abstract neonatal meningitis etc.[1].
Escherichia coli are the most commonly present With increasing use of drugs, microorganisms
bacterium in the human intestine, which helps in are attaining resistance to commonly used
preventing the entry of pathogenic microorganisms. antibiotics, which leads to downfall of effectiveness
E. coli are non-pathogenic in normal conditions, but of conventional medicines and therefore, search for
if present in excess, will become causative agent of new antimicrobial agents has become necessary.
various diseases like urinary tract infection, Traditional medicines have been used for many
diarrhoea, vomiting etc. With increasing resistance centuries by a substantial proportion of the
of microorganisms to antibiotics, there is a shift of population of India [2]. The interest in the study of
choice from allopathic to ayurvedic and naturopathy, medicinal plants as a source of pharmacologically
where herbs and spices are very common active compounds has increased worldwide. It is
ingredients of medicines. Herbs and spices are recognized that in developing countries like India,
used in Indian recipes as they impart aroma and plants are the main medicinal source to treat
flavour to it. Most of the studies performed to check infectious diseases [3]. Approximately 20% of the
sensitivity of microorganisms for various herbs and plants found in the world have been subjected to
spices involve extraction of the active component(s) pharmacological or biological test, and a substantial
with some organic solvents. However, when these number of new antibiotics introduced in the market
plant products are used as ingredients of food, the are obtained from natural or semi-synthetic
effectiveness of extracts using organic solvents resources [4].
should not be a criterion. In the present study, The active ingredients of plants against
frequently used herbs and spices are selected, their microorganisms are mostly some of the secondary
extracts are made using distilled water and tested metabolites (i.e. alkaloids, glycosides etc.) that are
for its antimicrobial effect against E. coli the most present in abundance in herbs and spices
common intestinal non pathogenic organism. The commonly used in Indian food preparations. Herbs
antibacterial effect of various herbs and spices were are small plants used by human being for various
evaluated using various methods. All the herbs and purposes like medicines, food supplements for
spices tested were able to inhibit E. coli growth, but imparting flavour or scant, and as a part of offerings
Thyme (herb) and Turmeric (spice) were found to to God since beginning of civilization [5].
be most effective against E. coli. Spices have been defined as plant substances
from indigenous or exotic origin, aromatic or with
Keywords: Escherichia coli; antibacterial activity; strong taste, used to enhance the taste of foods [6].
turbidometry; Thymus serpyllum; Curcuma longa. Spices include leaves (bay, mint, rosemary,
coriander, laurel, oregano), flowers (clove), bulbs
1. Introduction (garlic, onion), fruits (cumin, red chilli, black pepper),
Food borne diseases have always been a major stems (coriander, cinnamon), rhizomes (ginger) and
concern in both developing and developed countries other plant parts [7]. The importance of spices can
[1]. Campylobacter jejuni, Staphylococcus aureus, be found not only in their flavouring, but also in their
Salmonella (over 1600 types), Escherichia coli medicinal, preservative and antioxidant properties
O157:H7, streptococci, etc. are some of the major [8]. Being plants, the natural foodstuffs, spices
bacterial species that causes food borne diseases appeal to consumers who tend to question the
[2]. E. coli are the most commonly found bacterium safety of synthetic additives [9, 10]. Antimicrobial
in the human intestinal tract. Under normal properties of spices have been documented in
conditions, its presence is conducive to digestive ancient literature and the interest continues to the
processes. But when present in excess or in virulent present [11-14]. However, little information is
form it causes diseases. Virulent strains of E. coli available emphasizing the preservative and
antimicrobial properties of spices [15].

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The aim of this study is to test the antibacterial for the turbidometric method. For paper disc and
effect of various herbs and spices on Escherichia Agar ditch method dense E. coli culture inoculated
coli. The herbs and spices used in traditional Indian in N- broth was used.
food preparations (Table 1) are selected for the
Turbidometric Method
study with the aim to spread the awareness of the
utility of herbs and spices among the masses with The inoculum was inoculated to N- broth with
the following objectives. different concentration of sample (20-600 µg/ml).
• Collection and identification of herbs and The tubes were incubated for 24 hrs at 37°C and
spices samples; optical densities were recorded at 540 nm. Each
• Extraction of samples using distilled water; concentration of all the samples was analyzed in
• Optimization of Inoculums concentration (E. duplicate along with the controls and their mean is
coli); used to calculate % inhibition, which is plotted in the
• Standardization and antimicrobial assay of graph against concentration.
herbs and spices using various methods (i.e. Paper Disc Method
Paper disc method, agar ditch method,
Turbidometry). The inoculum (100µl) was spreaded uniformly in N-
agar plates with the help of glass spreader and kept
Table 1. List of herbs and spices used along with their for five minutes. Pre- sterilized paper discs were
active ingredients dipped into different samples (herb and spice
extracts) placed in inoculated plates. The plates
Common Scientific were incubated for 24 hrs at 37°C and size of clear
Components Present
Name Name zones developed surrounding each disc was
Thymus measured by scale to the nearest mm and were
Thyme thymol
vulgaris
plotted in the graph.
Origanum
Oregano carvacrol
Vulgare Agar Ditch Method
Ocimum eugenol (1-hydroxy-2-
Tulsi With the help of cork borer four ditches were made
tenuiflorum methoxy-4-allylbenzene)
phenylpropanoid-derived per plate in the pre inoculated N-agar plates.
Zingiber Different extracts were added to these ditches
Ginger compounds, particularly
officinale
gingerols and shogaols (100µl) and plates were incubated at 37°C. After 24
Curcuma hrs diameter of clear zone produced surrounding the
Turmeric Circumin
longa ditches were measured to the nearest mm with the
eugenol, cinnamaldehyde, help of scale and were presented in the graph.
Cinnamomu
Cinnamon beta-caryophyllene, linalool,
m verum
and methyl chavicol. Statistical Analysis
phenylpropanoides such as
Syzygium The data of mean % inhibition were subjected
Clove carvacrol, thymol, eugenol
aromaticum Single factor analysis of variance (ANOVA) and
and cinnamaldehyde
Ferula unpaired t- test using variance. The Level of
Asafoetida ferulic acid significance of difference and calculated values are
asafoetida
discussed.

2. Materials and Methods 3. Results and Discussion

Preparation of samples Traditionally, herbs and spices are part of routine
Indian food preparations as they make food
The leaves (1 g) of the various herbs were overnight appealing by providing better appearance, smell and
soaked in water and ground with mortar and pestle taste. As mentioned in ‘Atharva veda’ these herbs
along with 5 ml of distilled water. The samples were and spices have healing, soothing and rejuvenating
then centrifuged (5000 RPM, 10 min) and the properties. Some of the common herbs and spice
supernatants obtained were used as an extract. used are chilli, clove, coriander, mint, garlic, ginger,
Fresh samples of the spices were ground with 5 ml turmeric, tulsi, cinnamon, tamarind, asafoetida etc.
of distilled water and centrifuged at 5000 RPM for ‘Ayurveda’ the indigenous system of Indian
10 min. The supernatant was separated and the medicine, uses a large number of spices in
samples were washed twice using distilled water. combinations as preventive and curative medicines
The pooled supernatants were used as sample [16].
(extract) and represented as µg/ml. There are number of reports available in
Preparation of inoculums (E. coli culture) literature stating the sensitivity of various bacteria
towards these herbs and spices used in food
A loop full of pure culture is inoculated in saline and preparations [17]. Most of these studies involve
the optical density is adjusted to 0.1 using extraction of the active component in the herbs and
spectrophotometer. This saline suspension is used spices using organic solvents. But as an ingredient

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of food, the antibacterial properties using organic cough. It is used in counteracting intestinal
solvent extraction should not be a criterion; flatulence by increases the levels of detoxifying
therefore, in the present study the commonly used enzymes in the body [20]. Ginger is commonly used
herbs and spices extracts are made with distilled to increase digestion, treat joint pains, common cold
water and tested for its antimicrobial effect against and throat infections and is also known to boost
E. coli. immunity [19]. Turmeric is a common ingredient
There are various direct and indirect methods present in most Indian food preparations. It is a
used to enumerate concentration of microorganisms known fact that turmeric prevents Alzheimer’s
from variety of samples [18] like microscopic count, disease which is supported by the fact that Indian
total count, spread plate for viable count etc. In the population has a lower percentage of people
present study three of the methods were used to suffering from this disease as compared to western
check antimicrobial activity of various spices and countries [20]. Cloves are used in Ayurveda,
herbs and to have an idea about most suitable Chinese medicine and Western herbalism. Cloves
method for the similar kind of analysis. Viz. are used as a carminative, to increase hydrochloric
Turbidometric method, Paper disc method and Agar acid in the stomach and to improve peristalsis [21].
ditch method. It is also used in dentistry where the essential oil of
Samples were analysed using turbidometric clove is used as anodyne for dental emergencies
method and mean % inhibition in growth of E. coli [22, 23].
against various concentrations (20-600 µg/ml) of
different spices is represented in Figure 1. All the
samples showed increase in % inhibition of growth
with increase in concentration. However, Turmeric
was found to be most effective in inhibiting growth
of E. coli even at 20µg/ml i.e. lowest concentration
used followed by clove and cinnamon. All the spices
showed statistically significant difference in %
inhibition (P< 0.05).

Figure 2. % Inhibition of E. coli when various

concentrations of herbs were tested using turbidometric
method

Figure 2 represents % inhibition in growth of E. coli

against various concentrations (20-600 µg/ml) of
Figure 1. % Inhibition of E. coli when various
concentrations of spices were tested using turbidometric
different herbs. All the three herbs tested showed
method increase in % inhibition of growth along with
concentration. Thyme was found to be most
Apart from antimicrobial activity, spices used for effective followed by oregano and tulsi. Unlike
present study has properties like analgesic, oregano and thyme, tulsi showed considerable
carminative, digestive, protective etc. Asafoetida is inhibition in E. coli growth only at higher
used in treating chronic bronchitis and whooping concentrations. Tulsi extracts are used in ayurvedic

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remedies for common colds, headaches, stomach Considering the above results, it can be
disorders, inflammation, heart disease, various suggested that addition of herbs and spices to the
forms of poisoning, and malaria [24]. Our results are food preparations helps to keep a check on the
contradictory to the literature According to which concentration Escherichia coli in the body. Tulsi and
tulsi is a plant with maximum antibacterial turmeric are used as most important food additives
compounds. Analysis of variance performed with antimicrobial activities in Indian recipes,
amongst different herbs showed statistically however, our results suggest that addition of thyme
significant difference (P<0.05) amongst herbs. A tea and oregano will also be helpful for the same.
made by infusing the herbs like tulsi, mint etc. in
water can be use for the prevention and cure of References
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