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Int. J. Pharm. Sci. Rev. Res., 30(1), January – February 2015; Article No. 41, Pages: 227-234 ISSN 0976 – 044X

Research Article

Snake Bite, Venom, Anti-Venom Production and Anti-Venom Activity of

Medicinal Plants: A Review
1 2 1 3
Rakesh Kant Kamal, Neha Sahu*, Jitin Rahul, [Link]
1
Department of Environmental Science & Engineering, Indian School of Mines, Dhanbad, Jharkhand, India.
2
Department of Biotechnology, KIIT University, Bhubaneswar, Odisha, India.
3
Department of Botany, D.B.S. College, Kanpur, Uttar Pradesh, India.
*Corresponding author’s E-mail: [Link]@[Link]

Accepted on: 09-11-2014; Finalized on: 31-12-2014.

ABSTRACT
Snakebite is a medically and socially significant issue in India, but it is one of the most neglected public health issues in poor rural
communities. The estimated death incidence due to snake bites is 1,25,000 per year globally, which signifies that death rates
associated with snake bites is a serious epidemiology. The current prospectus of treatment and reporting protocols need to be
upgraded to higher standards. Anti-venoms have been widely used for more than a century for treating snakebites and other
accidents with poisonous animals. Presently there are seven laboratories in India which produce anti-venom against four most
venomous and medically important Indian snake species- Cobra (Naja sp.), Krait (Bungarus sp.), Russell's Viper (Daboia russelii) and
Saw Scaled Viper (Echis carinatus sp.), the 'big four'. Most venom for anti-venom production in India is sourced from Chennai. In this
article, we review the production of venom and anti-venom in India and suggest areas of improvement, with the help of medicinal
plants capable enough to be used as anti-venom. In light of these observations, it is felt that there is a need to prioritize the
betterment of venom and anti-venom production protocols, public education, and snake bite treatment in India.
Keywords: Snake bite, Snake venom, Snake Anti-venom, Anti-venom production, Anti-venom activity of medicinal plants.

INTRODUCTION There are four common venomous snakes found in India.

They are Common Cobra (Naja naja), Common Krait

S ince ancient times, snakes have been worshipped,

feared, or loathed. Unfortunately, snakes remain a
painful reality in the daily life of millions of villagers.
Indeed, although anti-venom is produced in sufficient
(Bunguraus caeruleus), Russell's Viper (Daboia russelii)
and Saw Scaled Viper (Echis carinatus)2 (Fig 2.). They are
known as the “Big Four” of India. Apparently they are not
the only species which are venomous, but are also found
quantities by several public and private manufacturers,
throughout India. They are widely distributed across the
most snake bite victims don't have access to quality care,
country. There are other venomous snakes like the King
both morbidity and mortality due to snake bites are high
Cobra (Ophiophagus hannah), Hump Nosed Pit Viper
(Fig 1.). The neglected status of snake bite envenoming
(Hypnale hypnale), Banded Krait (Bunguraus faciatus) are
has recently been challenged1 but as outlined below,
also found in India.
apart from the production of anti-venom, snake bite
envenoming in India shares all the characteristics of a
neglected tropical disease.
This review aims at summarizing and discussing the
epidemiology, clinical features, diagnosis, and treatment
of snake bite envenoming in India.

Figure 2: Characterization of Venom According To Snake

Family
Death rate associated with snake bites is one of the major
epidemiology in various regions of the world. About
Figure 1: Estimated Mortality Due To Snake Bites In India 35,000 to 50,000 people reportedly die of snake bite in
3
India every year especially in rural areas (Fig 3.).

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Int. J. Pharm. Sci. Rev. Res., 30(1), January – February 2015; Article No. 41, Pages: 227-234 ISSN 0976 – 044X

southern India confirmed that delayed anti-venom

administration was associated with an increased risk of
complications11,12. The bite victim should be reassured,
the bitten limb immobilized with a makeshift splint or
sling, and the patient transported. Walking is
contraindicated, because muscular contractions promote
venom absorption.
Anti-venom Serum Therapy
Immunotherapy is the only specific treatment for snake
bite envenoming. Anti-venoms are produced by
fractionation of plasma obtained from immunized
Figure 3: Statistics of Deaths by Snake Bites animals, usually horses13. They can be either monovalent
Deaths due to snake bites may be reduced up to a certain or polyvalent, depending on the number of species (single
extent by the use of proper first aid and medical support or multiple, respectively) whose venoms are used for
in the form of anti-venom. Despite increasing knowledge immunization.
of snake venom's composition and mode of action, good Although monovalent anti-venom has often been
understanding of clinical features of envenoming and considered more efficacious, the production of polyvalent
sufficient production of anti-venom by Indian anti-venom is preferred in many countries as snake
manufacturers, snake bite management remains species identification is generally not possible for the
unsatisfactory due to various downfall scenarios. Anti- attending physician.
venom’s use may lead to significant side effects, anti-
venom development is time consuming, expensive, and Anti-venoms have been available in South Asia for the
requires ideal storage conditions4 and therefore search past 60 years, and all existing products are manufactured
for venom inhibitors, either synthetic or natural, that by Indian companies.
could complement or substitute for the action of anti- Traditionally, the production has focused on four species
venoms are of greater importance. believed to be responsible for most deaths: N. naja, B.
Snake Bite caeruleus, D. russelii, and E. carinatus. However, a
number of other species that contribute to morbidity and
A snake bite is basically an injury caused by a snake, often mortality in the region have not been considered, and
resulting in puncture wounds inflicted by the animal's envenoming by these species usually does not respond
fangs and sometimes resulting in envenomation. adequately to existing anti-venoms14-16.
Although majority of snake species are non-venomous
and typically kill their prey with constriction rather than Snake Venom
venom, venomous snakes (15% out of 3000 known Snake venom is essentially highly modified saliva that is
species)5-7 are reported to be found on every continent made up of 90% proteins (by dry weight) and most of the
except Antarctica5. proteins are enzymes17. Most of these enzymes are
Snake Bite Management harmless to humans and are generally not dangerous
when ingested.
Health workers in rural districts are usually poorly trained
to manage snake bite envenoming, which is a complex There are about 20 toxic enzymes known to man and
emergency. A recent survey conducted in India and unique mixture of these zootoxins and proteins, form the
Pakistan showed that many doctors were unable to lethal weapons of snakes. The venom contains
8 phosphodiesterases (attacks the cardiac system),
recognize systemic signs of envenoming . Another study
in northwest India revealed that most snake bite victims cholinesterase (loss of muscle control), hyaluronidase
presenting at primary health centres received inadequate (increased tissue permeability), ATPases (disrupts energy
doses of anti-venom and that out of 42 patients who fuel use) and various toxins (neurotoxins, cardiotoxins,
9 blood clotting toxins, bleeding toxins) and other enzymes
required assisted ventilation, only one was incubated .
a major components, as well as small peptides, amino
There are two important aspects of snake bite acids, carbohydrates, lipids, nucleosides, biological
management. amines, metal ions and proteases28. It is stored in a large
Proper First Aid sac like structure, known as the alveoli behind the
animal’s eyes and it is ejected through a set of tubular
Most experts agree that snake bite victims should be fangs.
transported as quickly as possible to a medical centre
where they can be clinically evaluated by qualified Fresh snake venom is neutral or weak acid, and it is
medical staff, and where anti-venoms are available. In alkaline when it is placed for a long time, and on exposure
fact, time of transport was shown to be a crucial to air fresh venom produces foam and will be non-
determinant of snake bite mortality10 and studies in venomous and putrid when kept at room temperature for

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Int. J. Pharm. Sci. Rev. Res., 30(1), January – February 2015; Article No. 41, Pages: 227-234 ISSN 0976 – 044X
18
24 hours (toxicity disappear following UV irradiation and Myotoxic Venoms
heat treatment; dealt with formaldehyde the toxicity also
This venom is found in Bothrops moojeni, commonly
disappear but antigenic property is retained).
known as the Brazilian lancehead snakes. It is known to
Classification of Snake Venom cause muscular necrosis. Its symptoms are a thickened-
tongue sensation, dry throat, thirst, muscular spasms and
Snake venom can be broadly categorized into many
convulsions. It also causes the stiffness of the jaw, neck,
types, but the most considerable types are:
trunk and limbs along with severe pain in movement24.
Hemotoxic Venoms The victims often start with drooping eyelids and then
turn to more severe results like loss of breath and
These attack the cardiovascular system, circulatory
blackish brown urine discharge. Mitotic venom contains
system and muscle tissues, thus directly leading to heart
19 peptides that destroy the muscle fibre proteins and result
failures . Normally, neither pain nor any other symptoms
in my necrosis (muscle destruction). In the very later
can be celebrated for almost 1 - 3 hours (sometimes even
stages (when treatment is delayed) of the spread of this
8 hours). This makes it deadlier, as the victim is usually
venom, the muscle proteins enter the blood stream. The
beyond medical help, by the time the cause is even
kidney overworks in trying to filter out the toxins
ascertained. The effects can be seen as lethargy,
eventually causing kidney failure which ultimately is the
headaches, nausea, vomiting, etc. The scariest
reason for the dark coloration of urine24.
observations of the outcome of snakebite of this kind are
bruising or blood spots beneath the victim’s skin. In Venom Collection
extremely bad cases, blood is known to ooze out from all
In India, the Wildlife Protection Act provides in-situ
possible bodily openings. It is these venoms that usually
protection for all snakes and as such, snakes cannot be
cause excessive scarring, gangrene and permanent or
collected for venom extraction or any other purposes
temporary loss of motor skills. Worst cases can even
without prior permission of the state wildlife
result in the amputation of the affected limb20.
authorities25. There is no scientific study that adequately
Neurotoxic Venoms quantifies snake abundance (through the export of up to
10 million snake skins per year in the 1960s gives some
They go after the central nervous system and brain. They
indication), which has resulted in a conservative stance by
often result in respiratory paralysis and heart failures.
the wildlife authorities in some states and a general
Their effect can range between mild seizures to death21.
reluctance to permit capture of large numbers of snakes
Cobras, mambas, sea snakes, kraits and coral snakes are
for venom extraction to produce Antivenins or Anti-
known to possess it. The king cobras (Ophiophagus
venoms.
hannah) are the most infamous carriers of this venom.
Neurotoxic venom is essentially nerve destroying. Hence, Snake Anti-Venom or Antivenin
one can see speech and swallowing difficulties, drooling,
Anti-venom first developed by Calmette (1895) aimed to
difficulty in breathing, respiratory arrests, convulsions
neutralize venom toxins and was experimented against
and sometimes even prolonged unconsciousness in the
Indian Cobra (Naja naja). Anti-venom (or antivenin or
victims. The milder symptoms are dizziness, tunnel vision,
antivenin) is a biological product used in the treatment of
blurred vision and increased sweating. It causes a very
venomous bites or stings. It is a purified fraction of
fast degeneration of the synaptic nerves and this is the
immunoglobulin or immunoglobulin fragments
reason for the blockage of nerve impulses sent to and
fractionated from the plasma of animals that have been
from the brain to the muscles21.
immunized against a venom (venom from: snake, spider,
26
Cytotoxic Venoms scorpions etc) .
This is a milder form that generally causes only localized Anti-venom Types
symptoms at the location of the bite. This is a cell
Anti-venoms can be classified into monovalent (when
destroying poison that destroys everything in its path -
they are effective against a given species’ venom) or
blood vessels, cells and tissues22.
polyvalent (when they are effective against a range of
The symptoms of the invasion of this venom are generally species, or several different species at the same time)
seen around 10 - 15 minutes after the snake encounter. types.
The results are generally localized pain accompanied by Anti-Venom Production
severe swelling and bleeding. One can easily spot the
Anti-venom production started in 1890’s by Albert
formation of red blisters near the bite area. This venom
Calmette, who was living in present-day Vietnam when a
causes blue/black spotting due to limited blood
flood forced monocle cobras into a village near Saigon,
circulation. The body often revolts against the invasion of
where they bit at least 40 people and killed four. Inspired
this venom by causing nausea and vomiting. If this is not
by the then-new science of vaccinations, by 1896
treated within four hours, it generally needs an
23 Calmette had discovered the process of injecting horses
amputation .
with venom until they produced antibodies, taking the

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Int. J. Pharm. Sci. Rev. Res., 30(1), January – February 2015; Article No. 41, Pages: 227-234 ISSN 0976 – 044X

serum out of their blood and injecting it into snake-bitten Other animals which can be used for the purpose of
humans as anti-venom. Immunization are Rabbit, Donkey, Goat and Sheep.
Basic Protocol for Antivenin Production Immunizing
The first step in anti-venom production is milking venom  A particular amount of venom along with distilled
from the desired snake, spider or insect. The venom is water, buffer solution (0.2 M Tris HCl) and adjuvant
then diluted and injected into a horse, sheep or goat. The (a substance which enhances immune response e.g.
subject animal will undergo an immune response to the Nanostructured silica, cobalt-60) is measured and
venom, producing antibodies against the venom’s active injected into the horse.
molecules which can then be harvested from the animal’s
blood and used to treat envenomation27. Internationally,  The amount of venom to be injected is divided into
antivenins must confirm to the standards of small volumes and injected separately into different
pharmacopoeia and the World Health Organization organs where antibodies are produced (back of the
(WHO)28. neck - lymph nodes) to prevent ulcer or sore skin and
maximize the area of immune reaction.
The basic protocol for anti-venom production includes
following steps:  The process of immunizing depends upon

Milking the Venom  Type of anti-venom

 The first step involves transferring of the captive or  The snake used
quarantine snake into a clean milking room.  The sort of antibodies desired
 Next the snake is grabbed with the thumb and the Purification
index finger at the very back of its head, where the
venom glands reside and the venom glands are Blood is taken from the immunized animal and is
pressed to ejaculate the venom through a plastic or centrifuged to separate plasma from the blood cells.
rubber film into the vial (glass ware). Basically, it is the plasma which contains effective
Cooling Down and Labeling antibodies against the venom.
 After milking the venom, the venom is cooled to The plasma is the filtered and remaining blood cells may
below -20o C and freeze dried (lyophilized) for easier be injected back into the animal.
storage and transport. This will concentrate the
The anti-venom thus produced is required to be
venom and remove the water.
separated from unwanted proteins. This is done though:
 Labeling of the venom from one particular species
 Precipitation- The precipitate contains unwanted
should be done because the anti-venom produced
amino acids and proteins, which are then discarded
from particular venom is specific for it.
off.
 It also prevents from being mixed with the anti-
venom produced from the sub species of the same  Adjusting the Plasma’s pH to 7.4- This step is
species. required to maintain the neutral pH of the solution
containing plasma.
 The label should contain the following information:
Specificity of anti-venom, plasma unit number and  Adding salts (ammonium sulphate, hydroxylapatite
date of collection. etc) - Salts stabilizes the solution.

Choosing an Animal for Immunization Breaking down of antibodies into small parts isolates its
active ingredients consisting of the required anti-venom.
Immunization is the process by which an individual’s
immune system becomes fortified against an agent The anti-venom should be deemed safe and effective by
(immunogen). In this context the selected animal is the Food and Drug Administration.
allowed to produce antibodies against the specific Human Use
immunogen (snake venom).
 The purified anti-venom is lyophilized and
Generally, horse (Equus) is the preferred animal: refrigerated in vials.
 They thrive in many environment worldwide  In case of emergency the vials are filled with saline
 They have a large body mass solution and injected intravenously (near the bitten
region).
 They have long lives
 A patient envenomed by snake bite requires 25-30
 Comparatively big veins and friendlier
vials of anti-venom to be healed (1 vial = 6000
 Easy to handle antivenin units).

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Int. J. Pharm. Sci. Rev. Res., 30(1), January – February 2015; Article No. 41, Pages: 227-234 ISSN 0976 – 044X

precipitation, if not stored properly.

 Must be preserved always as freeze-dried sample.
 Anti-venom is unable to reach in remote areas.
 Inefficiency in reaching some countries due to
storage difficulties.
 Resource intensive, pain staking.
 Complications in identifying the type of anti-venom
to be used.
 Allergic reaction to certain individuals.
 Non-availability of anti-venom in hospitals.
Side Effects of Anti-Venom
Side effects of anti-venom therapy are anaphylactic
reaction (difficulty in breathing and swallowing; hives;
Figure 4: Flowchart: Fractionation of Plasma for
itching especially of feet or hands; reddening of skin,
Purification of IgG
especially around the ears; swelling of eyes, face, or
Stability and Storage inside of nose; unusual tiredness or weakness), serum
sickness (enlargement of the lymph glands; fever;
 Stability is essential to determine the shelf-life of the
generalized rash and itching; inflammation of joints),
product and intends to prove that anti-venom
pyrogen reaction - probably due to the action of high
remains stable and effective.
concentrations of non-immunoglobulin proteins present
 Quality control parameters determined at regular in commercially available hyper immune venom19.
time intervals are venom neutralization potency,
Other Applications of Snake Venom
turbidity and content of aggregates.
 Treatment of Breast Cancer
• Anti-venom is stored at temperature within a range
that assures the stability.  Treatment of Blood Clots
• For liquid preparations, requires storage temperature  Used as Morphine and Anaesthesia
at between 2˚ and 8˚ Celsius.
 Treatment of Alzheimer’s Disease
• Interruptions in temperature may lead to
Snake venoms are used to control heart diseases, high
deterioration.
blood pressure, cancer (contortrostain produced by
Anti-Venom Producing Centres in India Agkistrodon contortrix - is cytostatic in nature and found
to lower the growth of breast cancer in mice), tumor,
 Bengal Chemicals and pharmaceuticals Ltd.- Kolkata
polio, neurological disorders (enzymes from cobra venom
 Central Research Institute of Kasuli - Kasuli were found to cure Parkinson’s and Alzheimer’s diseases),
excessive bleeding (a blood clotting protein in Taipan
 Haffkine Biopharmaceutical Co - Mumbai
venom stop bleeding during surgery or after major
 King Institute - Chennai trauma), blood clotting (ancrod - obtained from Malyan
pit viper, used to develop angiotensin converting enzyme
 Vins Bio-products Ltd.- Hyderabad
inhibitors to treat stroke victims), severe allergies
29
 Biological ‘E’ Ltd.- Hyderabad amongst others . Other interesting areas of snake venom
Limitations of Anti-Venom include the treatment of viruses (as venom contains
phospholipidases which break down cell membrane),
 Cause various side effects. aging and some are even used in commercial wrinkle
30
 Cannot undo damage already caused by venom, so cream .
anti-venom treatment should be started as soon as Case Study
possible.
According to Leslie Boyer a physician and head of the
 Mostly administered intravenously but the route may Viper Institute at the University of Arizona, the process is
not be uniformly effective. much improved but the steps largely remains the same.
 Production is time consuming and expensive. Efforts are continuously being made to produce anti-
venom for the world’s deadliest snake bites. There’s one
 Liquid anti-venom may lose its activity due to protein other, quirkier way to make anti-venom–one that

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Int. J. Pharm. Sci. Rev. Res., 30(1), January – February 2015; Article No. 41, Pages: 227-234 ISSN 0976 – 044X

physicians don't exactly recommend. For decades Bill sarsaparilla Hemidesmus indicus could significantly
Haste, milked about 100 snakes a day with his bare neutralize lethality, haemorrhage, defibrinogenation,
hands, and in 1948 began injecting himself with edema, PLA2 activity induced by Daboia russellii venom. It
increasing doses of diluted cobra venom in order to also neutralized Naja kaouthia venom induced lethality,
develop his own immune resistance. At the time of his cardio-toxicity, neuro-toxicity and respiratory changes in
death (not caused by snakebite), he’d survived 172 bites experimental animals37. Beta-sitosterol and stigmasterol
from many of the world’s deadliest snakes, including a isolated from the root extract of Pluchea indica may play
blue krait, a king cobra and a Pakistani pit viper. He flew an important role, along with antiserum, in neutralizing
around the world to donate transfusions of his antibody- snake venom-induced actions.
rich blood to treat 21 snakebite victims. Venezuela made Several plant constituents like flavonoids, quinonoid,
him an honorary citizen after he traveled into the jungle xanthene, polyphenols and terpenoids possessed protein
to donate blood to a young snake-bitten boy. According binding and enzyme inhibiting properties and also inhibit
31
to his wife Nancy, all 21 patients survived . snake venom phospholipase A2 (PLA2) activities of both
Anti-Venom Activity of Medicinal Plants Viper and Cobra venom27. Triterpenoids present in V.
negundo and [Link] may involve in venom
The use of plants against the effects of snakes bite (Table
inactivation processes. The pentacyclic triterpenes (free
1.) has been long recognized; more scientific attention
or as glycosides) are found widely in several plants
has been given since last 20 years32. Extracts from plants
possessing snake antivenomic properties (Aegle
have been used among traditional healers, especially in marmelos, Centipeda minima, Aloe vera, Phyllanthus
tropical areas where there are plentiful sources, as niruri, Alstonia scholaris, Phyllanthus emblica,
therapy for snakebite for a long time. Several medicinal
Elephentopus scaber etc.) and provide nearly 20%
plants, which appear in old drug recipes or which have
protection against snake venom38. Eclipta prostrata is a
been passed on by oral tradition, are believed to be
tropical and subtropical plant used as anti-venom against
snakebite antidotes. In modern science, there have been
snakebite in China and in Brazil39. It was reported that the
many attempts to study these plants to clarify their
methanolic extract from Cordia verbenacea significantly
effectiveness. India has a rich tradition of the usage of inhibited paw edema induced by Bothrops jararacussu
medicinal plants. snake venom40. An active compound from the Strychnus
Many Indian medicinal plants are recommended for the nux vomica seed extract, inhibited viper venom induced
treatment of snakebite and their anti-venomic properties lipid peroxidation in experimental animals. Jantropha
were confirmed by many scientists. The methanolic root Caracas, Acacia leucophloea, Albizia procera, Madhuca
extracts of Vitex negundo and Emblica officinalis were indica and Ocimum sanctum plants are applied as an anti
tested for anti-venom activity. Both plant extracts were snake in Orchha wildlife sanctuary region of Tikamgarh
able to significantly neutralize the Vipera russellii and District, Madhya Pradesh, India14. Total six medicinal
Naja kaouthia venom induced lethal activity, both in vitro plants used in the treatment of snake bite reported in
and in vivo studies33. V. russellii venom-induced Taindol village, district Jhansi, region of Bundelkhand,
haemorrhage, coagulant, defibrinogenating and Uttar Pradesh, India41.
inflammatory activity was significantly neutralized by It may be concluded that evidence are now available to
both plant extracts34. Hemidesmus indicus root extracts establish the scientific background of the traditional use
effectively neutralized Viper venom induced lethal, of plants against snakebite. The anti snake venom plants
haemorrhagic, coagulant anticoagulant and inflammatory contain more than one compound (secondary
35
activity . The butanolic extract of Eclipta prostrata plant metabolites) that are responsible for venom
partially inhibited the hemorrhagic activity but displayed neutralization. Thus medicinal plants with anti-venom
very low antiphospholipase A2 activity and did not inhibit activity could be considered as an effective alternative to
proteolytic activity of Malayan pit viper venom32, 36. mammalian antibody production for the treatment of
Lupeol acetate isolated from the root extract of Indian snakebite envenomation.
Table 1: Some of the Most Needed Anti-Venoms
Common
Species Name Anti-venom(s) Threat
Name
America Lanceheads Bothrops species Antibotropico Polyvalent Responsible for 15 deaths per 100,000 people in Brazil
West African FAV-Afrique, SAIMR Echis anti- Most common snake danger in central and west sub
Africa Echis ocellatus
carpet viper venom Saharan Africa, up to 12.3% fatality rate (in Nigeria)
Haemato-Polyvalent snake anti-
Malayan pit Calloselasma Responsible for half of all hospitalizations in Malaysia,
Asia venom, Malayan pit viper,
viper rhodostoma where mortality rate reaches 18 per 100,000 people
Polyvalent anti snake venom
Australian Pseudonaja CSL brown snake anti-venom, CSL Responsible for half of approximately 200 snakebite
Oceania
brown snakes species polyvalent anti-venom deaths in Oceania each year

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Int. J. Pharm. Sci. Rev. Res., 30(1), January – February 2015; Article No. 41, Pages: 227-234 ISSN 0976 – 044X

CONCLUSION Products. 59, 1996, 664–667.

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for emergency handling is significant. The veracity of the 12. Mukherjee AK, Maity CR. Biochemical composition,
herbal assertions holds a good promise for the lethality and pathophysiology of venom from two cobras-
development of novel snake anti-venom drug in future. Naja naja and N. kaouthia. Comparative Biochemistry and
The combinations of herbal compounds with anti-venom Physiology, 131, 2002, 125-132.
serum may also be a good prospective as well as effective 13. Gutierrez JM, Lomonte B, Leon G, Rucavado A, Chaves F.
in neutralizing snake venom. Most importantly medicinal Trends in snakebite envenomation therapy: scientific,
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Madhya Pradesh, India. Journal of Botanical Research, 4(1),
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