evaluation of adaptogenic activity of fruit extracts of amla and ashwagandha by using microorganisms

1. EVALUATION OF ADAPTOGENIC
ACTIVITY OF THE FRUIT EXTRACT OF
Phyllanthus emblica AND ROOT
EXTRACT OF Withania somnifera BY
USING MICROORGANISMS
BY
SHAIK SHAHEENA
CH.DINESH BABU
UNDER THE SUPERVISION OF
N.KANAKADURGA DEVI M.PHARM.,Ph.D
KOMMAREDDY VENKATA SADASIVA RAO SIDDHARTHA COLLEGE OF PHARMACEUTICAL SCIENCES
2014

2. Any individual who can adapt easily to changing
situation(physical, mental, microbial, chemical) can stay
healthy
ADAPTOGENS was first coined by Dr.NIKOLAI
LAZAREV a Russian scientist only 50 years ago but
Indian Physicians practicing before 5000years-
RASAYANAS
It is sold in the market as Modern Nutraceuticals
with different labels

3. An ADAPTOGEN is a substance that
demonstrates a NONSPECIFIC
enhancement of the body’s ability
to resist a stressor
NON SPECIFIC – NOT CAUSED BY SPECIFIC AGENT
RESISTANCE – THE POWER OF RESISTING

4. PROPERTIES OF ADAPTOGENS:
•The substance is relatively Non – Toxic to
the recipient
•It has Non specific activity and increases
resistance of organism to broad spectrum
of adverse biological, chemical and
physical factors
•These substances tend to help, regulate
or normalize organ and system function
within the organism

5. HOW DO
THEY WORK
……….?

6. One theory suggested that Adaptogens function primarily
due to anti-oxidant and free radical scavenging activity
Most of the adpatogens are Anti-oxidants but substances
having Anti-oxidant properties ( Green Tea, Rosemary,
Cranberry) are not sufficient to make it an adaptogen.
Amphoteric is a substance that normalizes function of an
organ or a system within the body
All the herbs are Amphoterics but none of them qualify as
adaptogens.
So Adaptogens act as broad spectrum amphoterics to living
organisms.

7. Later research postulated that adaptogens work primarily by
affecting Hypothalamic/Pituitary/ Adrenal (HPA) axis and
Sympathoadrenal System (SAS)
Thus, adaptogens modulate our response to stress and help
regulate the interconnected endocrine, immune and nervous
system
This re-regulation of a highly disordered stress system is
achieved by metabolic regulators such as cytokines,
catecholamines, glucocorticoids, cortisol, serotonin, nitric
oxide, sex hormones, corticotrophin releasing factor (CRF)
This broad array of biochemical activators explain why
adaptogens have Anti- Inflammatory, Anti-Oxidant, Anxiolytic,
Anti-Depressant or nervine effects as well

8. Adaptogens can help people handle stress by providing :
•Anti- Oxidant activity
•Liver protection and Anti toxin activity
•Improved blood sugar metabolism
•Less craving for alcohol and sugar
•Improved immune resistance
•Increased stamina and energy
•Improved muscle tone
•Increased strength
•Faster recovery
•Better focus and concentration
•Less anxiety
•Better sleep
•Better motivation and productivity
•A feeling of well being
•Better moods

9. Stress induced health problems:
•Angina
•Asthma
•Auto immune Diseases
•Cancer
•Cardiovascular disease syndrome
•Common cold
•Depression
•Diabetes( Adult onset type II )
•Headaches
•Hypertension
•Immune suppression
•Irritable Bowel Disease
•Menstrual irregularities
•Pre menstrual tension
•Rheumatoid arthritis
•Ulcerative colitis
•Ulcers

10. The plants with Adaptogenic activity is
1. Red ginseng
2. American ginseng
3. Siberian ginseng
4. Ashwagandha
5. Astragalus
6. Liquorice
7. Tulsi
8. Satavari
9. Amla
10.Ginger

11. Ashwagandha Root ( Withania somnifera)
This herb is one of the most prominent Rasayana ( rejuvenative )
remedies of Ayurveda
It is one of the few calming Adaptogens and has traditionally been used
for Anxiety, Bad Dreams, Insomnia and Nervous Exhaustion
It acts as Anti spasmodic and Anti Inflammatory, in treating Fibromyalgia,
restless leg syndrome, mild Tourette’s Syndrome and Osteo arthritis
It is an immune amphoteric useful for hypo and hyper immune conditions
It enhances male fertility (sperm count and sperm motility)
Due to its Iron content, it benefits iron deficient anemia
Ashwagandha also stimulates thyroid function

12. Amla :
Shining through the Three Stages of Stress
It is one of the most used herbs in the world as
it is a part of many formulas such as Triphala,
consumed by millions daily with their meals as
delicious and digestive condiment

13. The general aim of the Adaptogen therapy lies in their
ability to reduce stressful agitation reactions during the
ALARAM PHASE of the stress response
Support the physiological rallying during the
RESISTANCE PHASE of stress response
And prevent or at least delay the state of fatigue during
the FINAL STAGE of the stress response
Doing so they provide a certain level of protection
against long-term stress and eventual exhaustion and
systemic decay

14. Amla supports all the three stages of
stress reactions and is shown in tests to
be an Anti- Stress Immunomodulating
Adaptogen that supports us in emotional
and cognitive wellness
Amla provides a healthy balance
between the aggressive factors of
digestion ( the acids and enzymes) and
the protective factors of digestion ( the
mucous, structural cells and alkalinity)

15. AIM OF THE WORK:
TO EVALUATE ANTI-STRESS ACTIVITY
OF CRUDE EXTRACT OF THE FRUIT OF
Phyllanthus emblica AND ROOT
EXTRACT OF Withania somnifera
USING MICROORGANISMS LIKE
Escherichia coli, Staphylococcus aureus
AND Lactobacillus

16. PLAN OF THE WORK:
1.By subjecting the microorganisms
to stress conditions like exposure
to UV light in the presence of an
extract
2.Testing of survival of
microorganisms even after
subjecting to stress by COLONY
COUNT METHOD

17. PREPARATION OF CRUDE EXTRACT OF AMLA FRUIT:
The fruits are washed
thoroughly to remove
impurities and pulped
Then passed through 1/8”
sieve followed by 1/16” sieve
to obtain fine particles
The product is pasteurized at
0
80-85
C and sterilized for
storage

18. PREPARATION OF CRUDE EXTRACT OF ASHWAGANDHA ROOT:
10 grams of root powder is dissolved in
1000 grams of distilled water
0
C for 6 hours
Boil at 100
Store at room temperature and it is the
crude extract of roots of ashwagandha

19. Prepare nutrient agar
medium
Then prepare fresh
subcultures of the required
strains of microorganism

20. Prepare agar
plates by
using pour
plate
method
Allow the plate to
dry for
approximately 5
minutes
Allow the
discs to soak
in fruit extract
of Amla/ root
extract of
Ashwagandha
Using flamed
forceps gently
press each
disc to agar to
ensure disc is
attached to
agar
Let the plates be in
refrigerator for 10
minutes and incubate
the plates at 370C for
24 hours
After
incubation
expose the
plates to UV
light for
15minutes and
then Incubate
again for 24
hours at 370C

21. E.coli S.aureus
Lactobacillus
FOR
FRUIT
EXTRACT
OF
AMLA

22. E.coli S.aureus
Lactobacillus
FOR ROOT
EXTRACT OF
ASHWAGANDHA

23. EVALUATION OF ANTI-STRESS ACTIVITY:
Following aseptic conditions from each agar plate transfer two loop full
of sample around the disk into 10 ml of normal saline solution
From that serial dilution was performed for all three strains of Amla
fruit extract and Ashwagandha root extract
ASHWAGANDHA
E.coli
S.aurues
Lacto
bacillus
AMLA
E.coli
S.aurues
Lacto
bacillus

25. COLONIES ARE COUNTED USING QUBEC COLONY
COUNTER

26. EXPERIMENTAL RESULTS:
No of colonies at 106 dilution
S.NO TYPE OF EXTRACT E.COLI S.AUREUS LACTOBACILLUS
1 AMLA 128 112 72
2 ASHWAGANDHA 144 172 168
NUMBER OF CELLS PER MILLI
METER OF ORIGINAL SAMPLE =
NUMBER OF COLONIES ×
DILUTION

27. NUMBER OF CELLS PRESENT AT 106 DILUTION:
S.NO
TYPE OF
EXTRACT
E.COLI S.AUREUS LACTOBACILLUS
1 AMLA 1.28X108 1.12X108 0.72X108
2 ASHWAGANDHA 1.44X108 1.72X108 1.68X108
A more elaborated results can be seen when number of
colonies are converted into number of cells
In the presence of E.coli, Ashwagandha shows an increase in
the activity by 11% to the activity of Amla
While there is a significant increase of 35% in the anti-stress
activity of Ashwagandha to Amla in the presence of S.aureus
In the presence of Lactobacillus there is a sharp rise in the
activity of Ashwagandha to Amla by an increase in 57%

28. COMPARISON OF CELLS PRESENT AFTER EXPOSURE TO STRESS IN
THE PRESENCE OF EXTRACTS OF AMLA AND ASHWAGANDHA
1.28
1.12
0.72
1.44
1.72
1.68
2
1.8
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
E.coli S.aureus Lactobacillus
amla
ashwagandha

29. CONCLUSIONS AND SUMMARY:
Ashwagandha and Amla both are said to have Adaptogenic activity
according to Ayurvedic medicines
The results show that even after exposure to stress conditions (UV
light) for a considerable period of 15minutes the microorganism
managed to survive
This was seen when a diluted sample of the organism in the
presence of both the extracts after exposure to stress produced a
good number of colonies
Though E.coli is said to be an organism that can resist heat, high
temperatures and other forms of stress like exposure to UV light
other organisms like S.aureus and Lactobacillus also showed
excellent results

30. NOVELTY OF THE WORK:
Normally we see that evaluation of adaptogenic activity of an
herbal formulation or an extract is carried out by determining its
effects on restrain stress (by immobilization of rats). The use of
albino mice or albino wistar rat is commonly seen.
The novelty of the work is the use of
microorganisms to evaluate the activity of
adaptogens in the herbal extract.
Microorganisms of different strains are used to evaluate the anti-microbial
activity of the Flavonoids present in the herbal
formulations but a procedure to evaluate anti-stress activity of
microorganisms in the presence of an adaptogen has never been
performed

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33. THANK YOU
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