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Available online at www.jpsscientificpublications.com
Life Science Archives (LSA)
ISSN: 2454-1354
Volume – 4; Issue - 1; Year – 2018; Page: 1248 – 1259
DOI: 10.22192/lsa.2018.4.1.2
©2018 Published by JPS Scientific Publications Ltd. All Rights Reserved
Review Article
ANTIMICROBIAL ACTIVITY OF SECONDARY METABOLITES IN AN
INDIAN SPICES – A REVIEW
M. Manigandan* and P. Saranraj
Department of Microbiology, Sacred Heart College (Autonomous), Tirupattur, Tamil Nadu, India.
Abstract
Spices are used majorly in a minor quantity in Indian foods. Traditionally spices are used in food as
flavouring, colouring, preserving agent. The Indian sub-continent is blessed with a varied climate each of its
state produces some spice or the other. Spices have showed numerous health benefits in preventing and
treating a wide variety of diseases such as cancer, aging, metabolic, neurological, cardiovascular and
inflammatory diseases. Spices are obtained from a variety of sources such as herbs, shrubs, plants, trees and
their products as leaves, fruits, seeds, outer layer of tree, barks, etc. The potential uses of spices are due to
their secondary metabolites and numerous phytochemical constituents. The bioactive compounds such as
Piperine, Eugenol, Cinnamaldehyde, Lignans, Linalool, Thymoquinones, Curcumin and Allicin are obtained
from the spices and condiments like Black Pepper, Cloves, Cinnamon and Nutmeg contain active
compounds. These act as natural preventive components of several diseases and represent as antioxidants in
body cells. The most important aspect of spices is it does not contain any of those toxic materials to human.
The present review paper contains the resources of spices with its secondary metabolites and its
antimicrobial effects.
Article History Received : 12.12.2017
Revised : 05.01.2018
Accepted: 20.02.2018
Key words: Spices, Antimicrobial effects, Health benefits and Bioactive compounds
1. Introduction
Spices are the substances, which upgrade
and enhance the flavours of food. Spices are
obtained from dried roots, barks and seeds utilized
entire or pounded, fueled and are sweet-smelling.
The flavor and aroma they give a superb touch to
sustenance things. Normally spices are a fixing
used to season a dish in the supper amid its
planning and toppings are for utilizing at the table
to improve the dish as every individual's tastes
lean toward. Indian spices and condiments have an
* Corresponding author: M. Manigandan
E.mail: [email protected]
uncommon effect in different courses in human
life on account of its one of a kind flavors, taste
and fragrance. A large portion of the flavors and
herbs have dynamic standards in them and
improvement of these through pharmacological
and preclinical and clinical screening would mean
development of significant open doors for
effective commercialization of the item. There is a
long revealing connection amongst herbs and
spices with the human culture, these spices and
toppings have been utilized to improve kind of
sustenance since antiquated circumstances. Spices
are very important and popularized since
M. Manigandan/Life Science Archives (LSA), Volume – 4, Issue – 1, 2018, Page – 1248 to 1259 1249
©2018 Published by JPS Scientific Publications Ltd. All Rights Reserved
antiquated circumstances. Spices are powerful in
people meds and nourishment safeguarding
operators. Indian kitchens are real clients of the
spices and herbs, and India is one of the biggest
purchaser, maker and exporter of spices and herbs.
Financially, the spices assume an essential part
(Deepali et al., 2015).
Spices were proven to possess medicinally
important components, in particular, antimicrobial
compounds. The spices are an imperative slice of
the human diet, which have been used for
thousands of years traditional medicine; a spice is
defined as a natural compound or a mixture of
natural compound that is extracted from the seeds,
fruit, flowers, traunses (skins, root, leaves) of
more than a few plants and added to food in order
to provide clear, taste, smell or flavor (Farrell,
1990).
Secondary metabolites are rich and present
broadly in plants. The real segments are tannins,
terpenoids, alkaloids, and flavonoids. The course
of these mixes from herbs, spices, and plant
separates has been demonstrated to have
antimicrobial properties against an expansive
reach of hurtful microorganisms. At last, an
expanded enthusiasm for the antimicrobial
properties of plant-inferred items has been
watched these days due to its potential uses as
other options to engineered additives. Auxiliary
metabolites from plants have turned out to be
moderately solid and could be utilized to broaden
the timeframe of realistic usability of
nourishments keeping in mind the end goal to get
the best sustenance wellbeing issues (Scalbert,
1991).
2. Antibacterial properties of Spices
Naturally occurring compounds from
spices have shown antimicrobial properties. The
main factors that determine the antimicrobial
activity are the type and conformation of the
spices. Spices are used as an ingredient that
increases the taste and variety of food (Bulduk,
2004). Consumers have likewise required for food
substances with long time span of usability and
nonappearance of danger of causing nourishment
borne ailments. This observation has put weight
on the nourishment business for liberal expulsion
of compound additives and reception of common
contrasting options to get its objectives concerning
microbial wellbeing. This occasioned in
expanding look for new known for use in
nourishment protection frameworks, which cover
modified environment pressing, joint impact of
under deadly techniques, substitute antimicrobial
mixes, amalgamation of straight (utilized as a part
of low levels) and substitutions antimicrobials
(Brull and Coote, 1999). This increases substantial
challenges, regularly since there is increasing
nervousness concerning the use of chemical
preservatives and replica antimicrobials to
inactivate or inhibit growth of pathogenic
microorganism.
Behera and Indira (2002) examined the
growth of spices export from India during the
period from 1995. The export of Indian spices has
grown at 7.94 per cent in terms of quantity and
17.64 per cent in terms of value through this
period. Similarly, the export of other items like
spices, oils and oleoresins, mint oil and curry
powder has suddenly amplified during the period.
The growth of spices importation into India during
the period was found to be 6.69 per cent in terms
of value, which was much below the increase rates
of Exports. It was found that India imports clove
in comparatively large quantities and nutmeg,
pepper and other spices in small quantities, mainly
from the European Union and USA (Khan and
Krishnakumar, 2002).
Phytochemicals or secondary compounds
of each spice has an unique flavor and aroma and
they are helpful to plants by guarding them against
all types of insects, fungi, pathogens, parasites and
consumers (Walker, 1994). Spices are used to
increase the taste, aroma and variation of food
(Ceylan, 1997). Aromatic plant materials devour
stayed utilized as a part of nourishment
arrangement and conservation. Treatment of
irresistible ailments is turning into a genuine
anxiety because of expanding obstruction against
anti-toxins among the pathogens is expanding at a
disturbing pace. The anti-infection agents may too
expend unfavorable consequences for the human
body like impact along the ordinary verdure and
M. Manigandan/Life Science Archives (LSA), Volume – 4, Issue – 1, 2018, Page – 1248 to 1259 1250
©2018 Published by JPS Scientific Publications Ltd. All Rights Reserved
hypersensitivity. Spices consume remained used
for not individual flavor and aroma of the foods
but also to deliver antimicrobial benefits
(Nanasombat et al., 2002).
Spices could underwrite sharpness
Restorative properties of medicinal plants are
mainly due to the presence of various chemical
compounds, i.e. secondary metabolites like
alkaloids, glycosides, flavonoids, saponins,
tannins, carbohydrates and cured extracts. The
usage of plants as a source of preparations for the
treatment of several diseases dated back to early
history and people have this old tradition. Therapy
infectious sicknesses initiated long before people
remained aware of reality of microbes. Infectious
diseases caused by bacteria, fungi, viruses and
parasites are immobile a major menace to public
wellbeing despite the unbelievable growth in
human medicine. Medicinal plants are of great
standing in the health of those and communities
(Edeoga et al., 2005).
3. Phytochemicals in Spices
Phytochemicals inimitable in the sternest
sense, among their concoction properties and their
utilization as medications went for conjunctivitis
by nearby individuals. The therapeutic plants are
valuable for recuperating and additionally to cure
of human illnesses as a result of the nearness of
phytochemical constituents. Phytochemicals are
normally mixing in the restorative plants, leaves,
vegetables and roots that have guard instrument
and shield from different maladies.
Phytochemicals are essential and auxiliary mixes
(Koperuncholan et al., 2010). Chlorophyll,
proteins and normal sugars are incorporated into
essential constituents and auxiliary mixes have
terpenoid, alkaloids and phenolic mixes (Gil et al.,
2002). Terpenoids comprises of vital
pharmacological exercises including against
growth, mitigating, antimalarial and restraint of
cholesterol combination. Spices can be added to
nourishments in a few structures as entire spices,
as ground spices, or as disconnects from their
concentrates.
Spices are known as one of the most
remarkable ingredients of the Indian cuisine.
Without the spices, the exotic flavors of the Indian
food don’t come out. The spices and the herbs are
also a prominent reason why the Indian food has
become so favorite among the people across the
globe (Dinda et al., 2015). So, in short, it can be
said that the Indian cuisine and the spices go hand
in hand as the traditional food items of India are
seasoned with a wide array of spices (Ignacimuthu
et al., 2006).
The Indian cooks use loads of seasoning in
different types of shapes and colors. Cinnamon,
black mustard seed, golden turmeric, cardamom,
chilies, ginger root is the common spices used to
cook the Indian dishes. Depending of the type of
dish like vegetable, chicken, fish or red meat, the
required spices are added during the cooking time.
The great thing about the spices is that different
flavors can be extracted from the same spice by
using different methods of using like grinding,
roasting, adding the whole spice or by combining
the spice with other spices (Cortés et al., 2014).
The most common spices used in Indian food are:
Carom Seeds: These are mainly used tempered in oil or dry roasted before
seasoning any dish. These are used to
prepare the breads in the northern part of
India.
Cloves: The cloves come up with strong, warm flavors. These are one of the prior
ingredients of the ―Garam Masala‖ and
also cooked in the ghee or oil to prepare
spicy dishes.
Mustard: Three main types of mustards are
used in the Indian cuisine, namely the
brown mustard, black mustard and white
mustard. This spice is used to bring strong
flavors in the dishes.
Cumin: This spice is also known popularly as jeera and quite popular because of
its earthy aroma. This spice is used in its
raw form to increase the spicy aroma of the
Indian dishes.
M. Manigandan/Life Science Archives (LSA), Volume – 4, Issue – 1, 2018, Page – 1248 to 1259 1251
©2018 Published by JPS Scientific Publications Ltd. All Rights Reserved
Indian cuisine is famous all around the
world for its great taste, colours and aromatic
spices. Sometimes those who experience Indian
dishes once find other food little bit tasteless.
Indian cuisine also enjoys the enviable reputation
of being very healthy and unique in taste. India is
a diverse country with followers of many religions
living together for centuries and so you will see
diversity in different recipes for cooking Indian
food. Indian cuisine offers a mind-blogging
variety of food. The Persian, Mughal and the
Greek influence truly reflects in numerous dishes
even today. It can be seen significantly in
Haryana, Punjab, Uttar Pradesh, Delhi and
Kashmir (Panpatil et al., 2013)
Some of the popular spices used in healthy
food recipes are cinnamon, cardamom, cumin,
mustard, coriander seeds, fennel, asafoetida,
turmeric, ginger etc. Each spice of Indian food has
dozens of different flavors which strongly depends
on the cooking method like whether they are
cooked in oil or ground or roasted. When cooked
in oil, it’s called Tadka and this technique is quite
different to Indian cooking. When spices are
cooked in oil they transfer their flavor to the hot
oil and anything cooked in this oil carry the flavor
of spices in tasty recipes (Newall et al., 1996).
A few spices are utilized as a part of the
type of ground and utilized for fragrance in Indian
dishes. These are utilized as entire and disposed of
amid eating. A few spices are simmered to
improve t he taste and embellish the primary
course dish. The spices and the herbs are
additionally a conspicuous motivation behind why
the Indian nourishment has turned out to be so
most loved among the general population over the
globe. Cinnamon, dark mustard seed, golden
turmeric, cardamom, chillies, ginger root is the
basic spices used to cook the Indian dishes.
Indians readily use spice in their food, teas, drinks
and even sweets not with these wonderful
aromatics for their exquisite flavor. The secret of
the masterly art of Indian cooking requires a
thorough knowledge of the properties of each
spice and its blend with other spices. So one can
say that the characteristic of each curry relies
entirely on the balance of herbs and spices that go
into its creations. Local influence distinguishes
curries from one region to another (Onyeagba et
al., 2004).
Indian spices can be mixed in specific
amounts to make Spice Mixtures used in daily
cooking. Some of the famous spice mixtures are
Garam Masala, Chat Masala etc. The composition
of the mixture might vary according to region but
the basic ingredients remain same. Many of the
formerly exotic Indian spices are becoming
increasingly familiar to the western cook and can
be easily found in any grocery store in the west.
The unique blend of spices for each dish makes
differentiation among the dishes and awards them
the special fragrance which is only due to the
perfect mixture of exotic Indian spices (Tyler et
al., 2002)
Indian cooking is eminent everywhere
throughout the world for the spices and sweet-
smelling kind of the sustenance. The different
Indian eateries in Washington DC plan mouth
watering indulgences and serve the sightseers and
additionally the neighbourhood individuals of the
region. The Indian dishes are uniquely described
by the utilization of different spices and herbs in
the arrangement of Indian food. Every Indian dish
must be set up with one of a kind method, implies
each Indian dish has its own strategy of
arrangement not quite the same as alternate dishes.
The Indian cooking is increasing greater
prominence because of the rich taste, outlandish
flavor and sound arrangement. Remembering the
mixed taste of Indian dishes a few Indians have
opened different sustenance indicates over the US
make Indian feel home in US. Such restaurants are
widely famous in US as the Indian food is very
much liked in US due to the spices used are good
for health (Ficker et al., 2003).
Each region has special culinary art and is
absolutely different from one another. India is the
largest food producer in the world. So it became
easy for India for maximizing exports of food
products to US or other similar countries. India
has huge scientific and research talent which is
increasingly used in developing new food products
which get success in US. India is a diverse country
M. Manigandan/Life Science Archives (LSA), Volume – 4, Issue – 1, 2018, Page – 1248 to 1259 1252
©2018 Published by JPS Scientific Publications Ltd. All Rights Reserved
possessing large geographical area and numerous
cultures. The geographical regions, climate,
religions, languages and customs all have great
effects on the country's various cuisines
(Thompson et al., 2002).
India's culture can be well reflected in its
cuisines from different parts. Different locations
possess different cultures and are famous for their
own cuisine that represents India to the world.
Indian food in Orlando is famous as it is a tourist
place and people from various other countries
come there and enjoys Indian food a lot. Spices
have been important to mankind since the
beginning of history. Several mythological
evidence including ―Epic of Gilgamaesh,‖ and the
―Bagavad Gita,‖ suggest their use for several
purposes. Because of their strong preservative
quality, spices were also used for embalming.
According to Ayurveda, they help to
maintain the balance of the body humors (Gupta et
al., 2013a). Besides these, spices have been used
to change the physical appearance of food. For
instance, pepper and turmeric changed the color,
appearance and the taste of food with many health
benefits. Ginger, nutmeg and cinnamon improve
digestion, considered good for spleen and sore
throats (Prasad et al., 2011). Unfortunately, this
beneficial effect of spices is not clinically proven.
However, traditional practices emphasize the
health benefits of spices. Eventually, recent
studies highlighted other biological functions of
spices, including antimicrobial, antioxidant, and
anti-inflammatory (Tajkarimi et al., 2010).
4. Importance of Spices phytochemicals in
Antibacterial activity
Phytochemicals, also referred to as
phytonutrients, are found in fruits, vegetables,
whole grains, legumes, beans, herbs, spices, nuts,
and seeds and are classified according to their
chemical structures and functional properties. The
terminology used to describe phytochemicals
(Flavonoids, Phenols, Flavonols, Flavanols,
Proanthocyanidins and Procyanidins) can be
confusing. Phytochemicals include compounds
such as salicylates, phytosterols, saponins,
glucosinolates, polyphenols, protease inhibitors,
monoterpenes, phytoestrogens, sulphides,
terpenes, lectins, and many more. The
Phytochemical Family Tree shows the major
groups of phytochemicals found in foods. Tens of
thousands of phytochemicals have been identified,
and researchers speculate that there are likely
many more they haven’t yet discovered in the
foods we eat. Though, the broadest groups of
phytochemicals, such as flavonoids, isoflavones,
or anthocyanidins, often are referred to as if they
were a homogenous group, the individual
compounds within each group have different
chemical structures, are metabolized differently by
the body, and may have different health effects.
Flavonoids are the largest, most varied,
and most studied group of phytochemicals. In fact,
more than 6,000 flavonoids that occur in plant
foods have been described. Plants typically
produce several phytochemicals that act as a
protective mechanism against environmental
stressors; the more environmental stressors, the
more phytochemicals a plant produces. As a
result, phytochemical content can vary with
growing conditions. There’s little information on
the average intake of phytochemicals among
Americans. However, it shouldn’t be surprising
that the intake of phytochemicals is higher among
people consuming the recommended amounts of
fruits and vegetables. Though, recommendations
suggest consuming a variety of fruits, vegetables,
and whole grains, for example, single foods often
account for the majority of the total intake of some
phytochemicals (Baliga et al., 2011).
Food spoilage refers to an irreversible
modification in which food becomes not edible or
its quality is compromised. Such changes can be
driven by different factors, either physical
(oxygen, temperature, light) and/or biological
(enzymatic activity and microbial growth).
Despite the current technologies available in the
production chain (for instance freezing,
pasteurization, drying, preservatives), it seems
impossible to eliminate completely the risk of
food spoilage (Gutierrez et al., 2009). Lipid
oxidation is one of the main issues of food
spoilage. Hence, food industries have applied
antioxidants such as butylated hydroxytoluene
M. Manigandan/Life Science Archives (LSA), Volume – 4, Issue – 1, 2018, Page – 1248 to 1259 1253
©2018 Published by JPS Scientific Publications Ltd. All Rights Reserved
(BHT) and butylated hydroxyanisole (BHA) to
prevent spoilage (Stoilova et al., 2007). However,
their safety is doubtful and consumers are
progressively demanding natural compounds. For
this reason spices represent a potent tool for the
food industry, thanks to their natural properties
(Hyldgaard et al., 2012). Indeed spices possess
antioxidant capacity, mainly due to the presence of
phenolic compounds. They exhibit antioxidant
property by scavenging free radicals, chelating
transition metals, quenching of singlet oxygen,
and enhancing the activities of antioxidant
enzymes (Rubió et al., 2013).
Various reports showed that the
CO2 extract of ginger had in vitro activity
comparable with that of BHT in inhibiting the
lipid peroxidation both at 37 °C and 80 °C.
Moreover, pimento and black pepper extracts
reduced the formation of acrylamide up to 75 %
and 50 %, respectively, in a model mixture
simulating heated potato matrix (180 °C for 20
min). Eugenol, the main component of pimento
essential oil, limited the formation of acrylamide
by 50 % (Ciesarová et al., 2008). Some other
studied Antioxidants are: Quercetine (dill),
Capsaicin (red chilli), Curcumin (turmeric),
Carvacrol (oregano, thyme and marjoram),
Thymol (oregano, thyme), Piperine (black pepper)
and Gingerol (Przygodzka et al., 2011;
Srinivasan, 2014).
In Nigeria, fruits like cucumber and spices
(ginger and cloves) are being underutilised as
there is no ready-to-drink juice produced from
these ingredients (as at the time of this study)
which contain some phytochemicals of health
benefits. These phytochemicals have been
considered to be of crucial nutritional importance
in preventing chronic diseases such as cancer,
cardiovascular diseases and diabetes (Aruoma,
2003). It has been discovered that regular
consumption of fruits, vegetables, herbs and spices
has always been associated with health benefits
(Liu, 2004).
Clove and ginger contain a wide variety of
biologically active, nonnutritive compounds
known as phytochemicals such as alkaloids,
flavonoids, tannins, phlabotannins, steroids,
saponins among others (Sheetal and Jamuna,
2009). These phytochemicals impart health
benefits beyond basic nutrition (Oomah and
Mazza, 2000) with the antioxidants having
potential to reduce the risk of several deadly
diseases in man (Agte et al., 2000). Naturally
occurring compounds, known as phytochemicals
(phyto means plant in Greek) are thought to be
largely responsible for the protective health
benefits of these plant-based foods and beverages,
beyond those conferred by their vitamin and
mineral contents. These phytochemicals, which
are part of a large and varied group of chemical
compounds, also are responsible for the color,
flavor, and odor of plant foods, such as
blueberries’ dark hue, broccoli’s bitter taste, and
garlic’s pungent odor.
Research strongly suggests that consuming
foods rich in phytochemicals provides health
benefits, but not enough information exists to
make specific recommendations for
phytochemical intake. This continuing education
course discusses the classification and
mechanisms of action of phytochemicals; explores
research concerning their role in the prevention of
cardiovascular disease, cancer, and diabetes as
well as neurological diseases such as Parkinson’s
and Alzheimer’s; and provides suggestions RDs
can share with clients for consuming
phytochemical-richfoods (Baliga et al., 2012).
5. Antimicrobial activity of Spices
Spice plants and essential oils extracted
from them have become important due to their
potential antimicrobial and stimulating effects in
the animal digestive system. The antimicrobial
effectiveness of mustard, clove, cinnamon and
their essential oils were reported for the first time
around 1880's. Antimicrobial effectiveness of
spices depend on the kind of spice, its composition
and concentration, type and concentrations of the
target microorganism, substrate composition, and
processing and food storage conditions. Spice
plants have been used traditionally as coloring
agents, flavoring agents, preservatives, food
additives and as well as antiparasitic,
antihelmintic, analgesic, expectorant, sedative,
M. Manigandan/Life Science Archives (LSA), Volume – 4, Issue – 1, 2018, Page – 1248 to 1259 1254
©2018 Published by JPS Scientific Publications Ltd. All Rights Reserved
antiseptic and anti-diabetic substances in many
parts of the world. In addition, they possess
biological activities such as that of antioxidants
and hypocholesterolemics (Panpatil, 2003).
The utilization of a few synthetic
compounds in sustenance and furthermore
utilization of a few anti-infection pharmaceuticals
has made microorganisms to create obstruction in
their populace. Spices have brilliant antibacterial
properties, in numerous nations individuals utilize
these alongside bubbled nourishment
arrangements which really lessen its antibacterial
properties (Pankaj et al., 2012). Spices, for
example, garlic, ginger, clove and cinnamon has
been utilized generally for both culinary and
therapeutic purposes. In India garlic has been
utilized to cure wound contamination and
nourishment deterioration (Daljit et al., 1999).
Little data is accessible on the additive and
antimicrobial part of spices and their oils and the
part of different segments of fundamental oils in
the conservation of ruin of nourishment. To
anticipate nourishment waste, compound additives
are utilized to keep the development of sustenance
ruining microorganisms in the sustenance business
(Sethi and Meena, 1997). There has been steady
increment looking for substitute and effective
mixes for nourishment protection went for an
incomplete or aggregate substitution of
antimicrobial compound added substances.
Likewise there has been increment of customers
about sustenances free or with bring down level of
compound additive on the grounds that these
could be lethal for people. In this manner, Plant
materials with antibacterial properties have a
conceivable application in nourishment protection.
A few investigations have demonstrated that has
garlic has antimicrobial impacts (Iwalokun et al.,
2004).
Garlic extract exhibits broad spectrum
antimicrobial activity against many genera of
bacteria and fungi at room temperature (Al-Amin
et al., 2006). Natural antimicrobial compounds in
spices were originated to have antimicrobial
activity whatsoever their natural defensive
purposes, natural harvests are a irritating source of
biologically active compounds that consume
arisen as the result of natural assortment (Ali
Aydin et al., 2007). Among the evaluated 500,000
plant species, be that as it may, just a little rate has
been researched for phytochemical. More than
90% of bacterial, parasitic, and plant species are
unmoving really taking shape to be investigated.
The first antimicrobial movement was the sort and
adaptation of the spice, volume utilized, kind of
microorganism, plan of the sustenance, pH
esteem, temperature of the earth, and proteins,
lipids, salts, and phenolic substances exhibit in the
nourishment. The knowledge of synergistic/
antagonistic (antimicrobial) effects of extracts of
the spices of Myrtaceae, Zingiberaceae and can
have vital suggestions in relation to the Indian
culinary ways as most of these spices are used in
combinations in various Indian food arrangements.
Therefore, it is important to study the
antimicrobial effects of these spices in their
specific bulk as well as in blends. Though several
spice extracts exhibited different inhibitory effect
against tested micro-organisms, comparable value
of minimum inhibitory concentration of was
perceived in incident of each specific spice extract
against all the tested microorganisms. This
exposed that the equal concentration of various
individual extracts was effective to different
magnitudes in inhibiting the growth of tested
microorganisms (Kota and Paladi, 2013).
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DOI: 10.22192/lsa.2018.4.1.2
How to Cite this Article:
M. Manigandan and P. Saranraj. 2018.
Antimicrobial activity of Secondary
metabolites in an Indian Spices – A
Review. Life Science Archives, 4(1): 1248 –
1259.
DOI: 10.22192/lsa.2018.4.1.2