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Antimicrobial Activity ofEuryale
ferox. Salisb:- A ThreatenedAquatic Plant of Kashmir
Himalaya
ByJavid Ahmad Parray
M.Phil Research scholar
Supervisor: Prof.(Dr.) Azra N. KamiliCo-Supervisor: Dr. Raies A. Qadri
P.G Department of Environmental Science
University of Kashmir
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INTRODUCTION
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Medicinal plants contain chemical substances which produce a definite
physiological action on human body and animal system.
Medicinal Plants
Phenols and
Mucilages
Flavnoids
Essential
oils TanninsGlycosides
Steroids
Chemical substances
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pharmacological
activities
Antimicrobial
Anti cancerous
Stimulatory
Anti-
inflammatory
Sedative
Spasmolytic
Secondary metabolites
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The main objective of the work was to determine theAntibacterial
and Antifungalactivity ofEuryale ferox.
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Study Area
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Source: Department of Geology and Remote Sensing, University of Kashmir.
Study Area- IRSP6-LISS III October Image of Mansabal lake
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PLANT
PROFILE
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Parts StudiedSeeds, Rhizome,
Petiole and Leaves.
Euryale ferox
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Domain Eukaryota
Kingdom Plantae
Subkingdom ViridaeplantaePhylum Tracheophyta
Sub-phylum Euphyllophytina
Infra-phylum Radiatopses
Class MagnoliopsidaSub-class Nymphaeidae
Super- order Nymphaeanae
Order Nymphaeales
Family NymphaeaceaeSub-family Euphorbioideae
Tribe Euphorbie
Genus Euryale
Species E. Ferox
Scientific Classification
Source: http://Plants.gov/java/Euryale f erox/Classification Servelet
http://zipcodezoo.com/Key/Plantae/Euphorbieae_Tribe.asphttp://zipcodezoo.com/Key/Plantae/Euphorbieae_Tribe.asphttp://zipcodezoo.com/Key/Plantae/Euphorbieae_Tribe.asp7/31/2019 Viva Final Prese 2
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Bihar Madhubani
English Fox nut
Kashmir Jewer
Punjabi Juwar
Hindi Makhana
Korean Ga-si-yeon-kot
China Qianshi
Japan Onibasu
Sanskrit Mukhauna, Padma
Vietnamese Khiem Thuc
Source: http: //www. herbalextractplus.com/Euryale.cfm
Different names of
Euryale ferox
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An annual aquatic herb with rhizomatous stems,
rhizome erect or repent and unbranched.
Leaves directly arise from rhizomes and are of 4-
10 cm in length with long petiolete.
Floating leaves are prickly on petioles andalong veins.
Leaf blade is abaxially dark purple and
adaxially green and is 1.3-2.7 mm indiameter and primary veins are prickly on
both surfaces .
Seeds are very hard, remains dormant for year.
Botanical Description
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China, Korea, Japan, Russia, Bangladesh,
India and grows wild in J&K and is an non
endemic threatened plant. (Khan et al.,
2000; Dar et al., 2002).
Distribution
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The plant has been traditionally used though out the
world to cure many diseases including chronic-
diarrhea, kidney problem, leucorrhea and hypo-
function of spleen. (Brown, 1995). used as an analgesic, aphrodisiac, astringent,
oxytonic and tonic in China (Duke et al.,
1985).
Leaves are used in the case of difficult parturition(Duke and Ayensu, 2003).
The plant is internally taken to treat vaginal-
discharge, impotence etc.(Brown, 1995).
Medicinal Uses
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In Indian traditional system of medicines the dry seeds
of the plant are being used as immuno-stimulant for
mothers after child birth with relatively poor immune
status (Puri et al., 2000). E. ferox also has been shown to enhance the activities of
superoxide dismutase, catalase and glutathione
peroxidase in V79-4 cells (Lee et al., 2002)
Seed of this plant have cardio protective properties
which may link with ability of makhana to induce
TRP32 and TrX-1 protein and to scavenge ROS
(Samarajit et al., 2006).
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Euryale ferox
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Seeds ofEuryale
ferox
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METHODOLOGY
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Methodology involved:Selection and collection of the medicinal
plant.
Solvent extraction. Selection of specific bacterial and fungal
strains.
Antimicrobial assay Qualitative phyto-chemical screening
METHODOLOGY
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Identification:Plant was identified at KASH,Department of Botany, University
of Kashmir under Acc. No.1015.
Collection: August -September 2008
Identification andCollection
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The powder form of different plant parts
(leaf, rhizome, petiole and seeds)
collected were subjected to hotextraction (Soxhlet extraction).
Solvents: Petroleum ether, Chloroform,
and Methanol.
Crude extraction of the
sample
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Soxhlet extractor
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Fifty grams of dried powder ofeach plant part (leaf, rhizome,
petiole and seed) were used.
Extracts collected were later driedand weighed and kept for further
process.
Whole plant
extraction
Preparation of Discs
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:
Preparation of mediaNutrient Agar Medium Dissolve 37.0 gm in 1000 ml of
distilled water
Nutrient Broth Suspend 8.0 gm in 1000 ml of
distilled water.
Muller Hinton Agar Dissolve 38.0 gm in 1000 ml of
distilled water
Muller Hinton Broth Suspend 21g of the powder in
1000ml of distilled water.
Blood Agar Suspend 40 g of the blood agar
base in 1000 ml of distilled water.
Sabourand Dextrose agar Dissolve 64 g in 1000ml of
distlled water
Preparation of Discs
Dissolve 200 mg of extract in 25 ml of solvent.
(1l=8g)
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Test Microorganisms
Gram Positive Bacterial Strains
Staphylococcusaureus I
Staphylococcus aureus II
Staphylococcus aureus III
Pnemococcus spp.
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E. Coli I
E. Coli II
Pseudomonas aeruoginosa I
Pseudomonas aeruoginosa II
Proteus vulgaris
Salmonella typhii
Salmonella typhimurium
Klebsiella pneumonia
Acintobactersp
Citrobacter fruendi
Shigella flexneri
Gram Negative Bacterial Strains
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Strains were provided by Bacteriological
and Mycological section ( Department ofMicrobiology, SKIMS Soura. Srinagar.
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Disc diffusion method: (Nutrient Agar , MullerHinton Agar and Saboarnd Dextroser agar,Hi
Media)
Kirby-Bauer (Bauer et al, 2002)
method was followed.
SENSITIVITY TEST FOR ANTIMICROBIAL ASSAY
Antimicrobial susceptibility Tests:
The primary purpose of antimicrobial susceptibility testing isto determine the potency of crude drug.
Sterile discs with antimicrobial agents are poured on microbialcultures and inhibition zone diameters (mm) around the discs are
measured as the potency of crude plant extracts.
Mixed culture of bacteria obtained from the polluted drain (that
was spread on the nutrient agar plate) and twelve extracts ( 3 ofeach plant parts) were tested for antibacterial activity.
Extracts which shows maximum activity ( methanol seed andmethanol leaf extract ) were than tested against specific bacterial and
fungal strains.
Dil i S ibili T
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Minimum inhibitory concentration(MIC)
Minimum bactericidal concentration(MBC).
Total activity (ml)
Dilution Susceptibility Tests
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S. aureusATCC.25923
E. coliATCC.25922
Pseudomonasaeroginosa
ATCC.27853
Proteusvulgaris*
Shigella
flxmeriATCC.12022
Salmonellatyphi*
StandardATCCBacterial Strains
Strains were provided by Bacteriological and Mycological section,
Department of Microbiology, SKIMS Soura. Srinagar.
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Medium
Muller Hinton Broth (Hi
Media ) (NCCLS, 1990)
i f i i f f
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Antimicrobial Conc.
mg/l
Volume of solution (ml) Distilled water (ml) Antimicrobial Conc.
mg/l
Final Conc. at 1:20
dilution(g/ml)
10000 1 9 1000 _
10000 0.1 9.9 100 _100 1 9 10 _
100 0.1 9.9 1 _
10000 1 0 10000 500
10000 0.512 0.488 5120 256
10000 0.256 0.744 2560 128
10000 0.128 0.872 1280 64
1000 0.64 0.36 640 321000 0.32 0.68 320 16
1000 0.16 0.84 160 8
100 0.8 0.2 80 4
100 0.4 0.6 40 2
100 0.2 0.8 20 1
10 1 0 10 0.5
10 0.5 0.5 5 0.25
10 0.25 0.75 2.5 0.125
10 0.125 0.875 1.25 0.06
1 0.625 0.375 0.625 0.03
1 0.313 0.687 0.313 0.015
1 0.156 0.844 0.156 0.008
(As per Philips et al ., 1991)
Preparation of dilutions of extracts for use
in MIC determination
Ph t h i l S i Q lit ti T t
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Phyto-chemical Screening Qualitative Tests
Alkaloids
Dragendorffs
test
Flavonoids
NaOHand conc.
HCl test
Phenols
Ferricchloridetest
Glycosides
Fehlingstest
SteroidsLieberman-
Burchards
test
TanninsFerricchloride test
Saponins
Frothingtest.
Physiochemical standards ofUnani
formulations- 1987
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By
Isolation of Phyto-constituents
Thin Layer
chromatography
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RESULTS
.
A i i bi l i i f E l f i
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GPC= Gram Positive Cocii
GNB= Gram Negative Bacillus
+ = Inhibitory activity
- = No activity
Pt = Petroleum ether extract
C = chloroform extract
M = methanol extract
Antimicrobial activity ofEuryale ferox extracts against
mixed culture of GPC & GNB bacteria.
Strains Inhibition Zone Diameter (mm)
Rhizome Petiole Leaf Seed
Pt C M Pt C M Pt C M Pt C M
Mixed culture of GPC& GNB bacteria.
_ _ _ _ _ _ _ _ + _ _ +
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Strains Inhibition Zone Diameter (mm)
Petroleum ether Chloroform Methanol
Mixed culture of
GPC & GNB
bacteria.
_ _ _
GPC= Gram Positive Cocii
GNB= Gram Negative Bacillus;
+ = Inhibitory activity
- = No activity.
Solvent Control
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GPC=Gram Positive Cocci
GNB= Gram Negative
Bacilli
M=methanol
Pt=Petroleum etherC=chloroform
SM=Seed methanol
SC=Seed chloroform
SP= Seed pet ether
LM=Leaf methanol
LC=Leaf Chloroform
LP=Leaf pet etherPM=Petiole methanol
PC=Seed chloroform
PP= Seed pet ether
RM=Leaf methanol
RC=Leaf Chloroform
RP=Leaf pet ether
FIG. 1a: Solvent extracts
ofE. ferox
FIG. 1b: Solvent Control
Plate1: Mixed culture of GPC & GNB
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Fig 2a :E. coli Fig 2b:Staphylococcus aureus
Plate 2: Effect of different concentrations of methanol seed extracts ofE.ferox on E.coli and Staphylococcus aureus
SI = 5 l and S11 =55 l (seed methanol extract)
Antibacterial activity of methanolic seed extracts of Euryale
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S.
No.
Strains Inhibition Zone Diameter (mm)*
160 g 320 g Antibiotics
01 S. aureus I 120.64 23 0.41 Erythromycin(25)
02 S. aureus II 120.81 18 0.70 Vancomycin (20)
03 S. aureus III 101.63 18 0.81 Cotrimaxozole (24)
04 Pnemococcus spp 151.29 20 0.81 Lenzolid (22)
* Results are mean diameter of zone of inhibition of three replicates followed by standard
deviation; Methanol was taken as negative control and was found resistant in all strains.
Antibacterial activity of methanolic seed extracts ofEuryale
ferox against Gram Positive Bacterial strains.
Antibacterial activity of methanolic leaf extracts of Euryale
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S. No Strains Inhibition Zone Diameter (mm) *
160 g 320 g Antibiotic
01 S. aureus I NA NA Erythromycin (25)
02 S. aureus II 90.00 151.28 Vancomycin (20)
03 S. aureus III 171.28 201.73 Cotrimaxozole (24)
04 Pnemococcus
spp
121.07 160.40 Lenzolid (22)
*Results are mean diameter of zone of inhibition of three replicates followed by standard
deviation; Methanol was taken as negative control and was found resistant in all strains
Antibacterial activity of methanolic leaf extracts ofEuryale
ferox against Gram Positive Bacterial strains.
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Gram Positive Bacterial Strains
Inh
ibitionzonesDiameter(mm)
Fig. 1: Comparative assessment of antibacterial activity of methanolic seed and leaf Extract ofE.
ferox against Gram Positive Bacterial Strains (320g)
Antibacterial activity of methanolic seed extracts of Euryale
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S. No. Strains Inhibition Zone Diameter (mm) *
160 g 320 g Antibiotics01 E .coli I 12 1.15 15 0.00 Ceftazidime (30)
02 E. coli II 16 2.16 22 1.63 Gentamycin (20)
03 Pseudomonas aeroginosa I 18 2.82 28 0.81 Amikacin (15)
04 Pseudomonas aeroginosa II 16 0.00 27 1.29 Gatifloxcin (20)
05 Acintobacterspp 10 2.16 14 1.15 Piperacilin (0)
06 Citrobacter freundii NA NA Vancomycin (22)
07 Proteus vulgaris NA 19 1.41 Cefixime
08 Shigella flexmeri 141.41 231.61 Ciprofloaxcin (29)
09 Klebsiela pneumonia NA NA Gentamycin (25)
10 Salmonella typhi 18 0.81 25 0.57 Imipenin (28)
11 Salmonella typhimurum NA 15 1.28 Gatifloxcin (0)
*Results are mean diameter of zone of inhibition of three replicates followed by standard
deviation; Methanol was taken as negative control and was found resistant in all strains.
Antibacterial activity of methanolic seed extracts ofEuryale
ferox against Gram Negative Bacterial strains.
Antimicrobial activity of methanolic leaf extracts of Euryale
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S. No. Strains Inhibition Zone Diameter (mm) *
160 g 320 g Antibiotics01 E. coli I 8 1.22 100.16 Ceftazidime (30)
02 E. coli II 13 0.42 200.41 Gentamycin (20)
03 Pseudomonas aeroginosa I 16 1.10 22 0.24 Amikacin (15)
04 Pseudomonas aeroginosa
II
17 1.24 21 0.81 Gatifloxcin (20)
05 Acintobacter spp 10 0.16 16 0.40 Piperacilin (0)
06 Citrobacter freundii NA NA Vancomycin (22)
07 Proteus vulgaris NA 10 0.00 Cefixime
08 Shigella flxmeri 12 0.74 18 0.21 Ciprofloaxcin (29)
09 Klebsella pneumoniae NA NA Gentamycin (25)10 Salmonella typhi 11 0.08 21 0.14 Imipenin (28)
11 Salmonella typhimurum NA 10 0.16 Gatifloxcin (0)
*Results are mean diameter of zone of inhibition of three replicates followed by Standard
deviation; Methanol was taken as negative control and was found resistant in all strains.
Antimicrobial activity of methanolic leaf extracts ofEuryale
ferox against Gram Negative Bacterial strains.
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Fig. 2: Comparative assessment of antibacterial activity of methanolic seed and leaf extract
ofE. ferox against Gram Negative Bacterial Strains (320 g)
InhibitionZonesD
iameter(mm)
Gram Negative Bacterial Strains
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Plate 3: Antibacterial activity of methanolic seed and leaf extract of
E. ferox against S.aureus I and Pneumococcus spp
Fig. 3a: S. aureus I Fig. 3b:Pneumococcus Spp.
SI=160 g, S2=320 g= Seed methanol extract. L1=160 g,
L2=320 g = Leaf methanol extract, Ab= Antibiotic; M= methanol
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Fig. 4a: S. aureus II Fig. 4b: S. aureus III
Plate: 4 Antibacterial activity of methanolic seed and leaf extract ofE. ferox against
S.aureus II and S.aureus IIISI=160 g, S2=320 g= Seed methanol extract. L1=160 g,
L2=320 g = Leaf methanol extract, Ab= Antibiotic; M= methanol
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Fig. 5a: P. aureoginosa I Fig. 5b:P. aureoginosa II
Plate 5: Antibacterial activity of methanolic seed and leaf extract ofE. ferox againstP.
aureoginosa I and P. aureoginosa II
SI=160 g, S2=320 g = Seed methanol extract. L1=160 g, L2=320 g = Leaf
methanol extract, Ab= Antibiotic; M= methanol
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Fig. 6a: S. typhi Fig. 6b: E. coli I
Fig. 6c:E. coli II
Plate 6: Antibacterial activity of methanolic
seed and leaf extract ofE. ferox against S.
typhi,E. coli IandE. coli II.
SI=160 g, S2=320 g = Seed methanol
extract. L1=160 g, L2=320 g = Leafmethanol extract, Ab=Antibiotic;
M=methanol
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Fig. 7a: Shigella flexnerii
Plate 7: Antibacterial activity of methanolic seed and leaf extract ofE. ferox against Shigella
flexnerii andAcintobacter spp.
SI=160 g, S2=320 g = Seed methanol extract. L1=160 g, L2=320 g = Leaf methanol extract,
Ab= Antibiotic; M=methanol
Fig. 7b: Acintobacterspp.
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Fig. 8a: Citrobacter freundii
Plate 8: Antibacterial activity of methanolic seed and leaf extract ofE. ferox against Citrobacter
freundii and Klebsiella pneumonia
SI=160 g, S2=320 g= Seed methanol extract. L1=160 g,L2=320 g = Leaf methanol extract, Ab= Antibiotic; M= methanol
Fig. 8b: Klebsiella pneumonia
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Plate 9: Antibacterial activity of methanolic seed and leaf extract ofE. ferox against Salmonella
typhimurium and Proteus vulgaris
SI=160 g, S2=320 g= Seed methanol extract. L1=160 g,L2=320 g = Leaf methanol extract, Ab= Antibiotic; M= methanol
Fig. 9b: Proteus vulgaris
Fig. 9a: Salmonella typhimurium
Minimum Inhibitory Concentration (MIC) of methanolic seed extract of
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S. No. Strains MIC MBC MIC
Index
Total
Activity
(ml)
Gt. 10 g/ml
MIC MBC
01 S. aureus ATCC.25923 64 128 2.0 593.75 32 128
02 E. coli ATCC.25922 128 256 2.0 296.87 128 500
03 Pseudomonas aeroginosaATCC. 27853
64 256 4.0 593.75 16 64
04 Shigella flxmeri
ATCC.12022
ND ND _ _ ND ND
05 Proteus vulgaris* 500 ND 76 256 500
06 Salmonella typhi* 256 500 1.95 148.43 ND ND
Methanol was used as negative control and exhibited no activity against tested organisms;
ND = Non detectable within the range tested; * = isolated Strains; Gt. = Gentamicin
MIC index = 4 (bactericidal) and = 4 (bacteriostatic)
Total activity =highest value= highest inhibitory activity
y
Euryale ferox against standard bacterial strains (g/ml)
.
Minimum Inhibitory Concentration (MIC) of methanolic leaf extract of
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S. No. Strains MIC MBC MIC
Index
Total
Activity
(ml)
Gt 10 g/ml
MIC MBC
01 S. aureus ATCC.25923 128 500 3.90 737.5 32 128
02 E. coli ATCC.25922 256 500 1.95 368.75 128 500
03 Pseudomonas aeroginosa
ATCC.27853
256 500 1.95 368.75 16 64
04 Shigella flxmeri
ATCC.12022
500 ND ND 188.8 ND ND
05 Proteus vulgaris* ND ND ND _ 256 500
06 Salmonella typhi* 500 ND 188.8 ND
Methanol was used as negative control and exhibited no activity against tested organisms;
ND = non detectable within the range tested; * = isolated Strains; Gt. = Gentamicin.
MIC index = 4 (bactericidal) and = 4 (bacteriostatic)
Total activity =highest value= highest inhibitory activity
y ( )
Euryale ferox against standard bacterial strains (g/ml).
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Standard Bacterial Strains
MIC(g/ml)
Fig. 3: Comparative assessment of MIC of methanolic seed and leaf extract ofE. ferox
against standard bacterial strains (g/ml)
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Fig. 4: Comparative assessment of MBC of methanolic seed and leaf extract of
E. ferox against standard bacterial Strains (g/ml)
Standard Bacterial Strains
MBC(g/ml)
g/ml)
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Fig. 5: Comparative assessment of Total activity of methanolic seed and leaf Extract ofE. ferox
against standard bacterial Strains (ml)
Totala
ctivity(ml)
Standard Bacterial Strains
Antifungal activity of methanolic leaf and seed extract of Euryale ferox.
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S. No. Fungal Strains Leaf extract Seed extract
200 g 400 g 200 g 400 g
01 Crytococcus neoformans NA NA NA NA
02 Aspergilus fumigates NA NA NA NA
03 Candida albicans 9 0.81 16 0.70 70.3 12 0.81
04 C. kruesie 8 0.48 100.28 70.3 9 0.00
05 C.Stealoidea 10 0.62 150.34 80.2 11 0.5
06 C. tropicals NA 150.34 NA 18 0.14
07 C. parapsilosis NA NA NA NA
08 Rhizopus spp NA NA NA NA
09 Pencillum notatum 10 0.81 150.60 80.2 19 0.84
*Results are mean diameter of zone of inhibition of three replicates followed by standard
deviation; Nystatin was used as positive control and Methanol as negative control in all
strains.
Antifungal activity of methanolic leaf and seed extract ofEuryale ferox.
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Fig. 6: Comparative assessment of antifungal activity of methanolic Seed and Leaf
extract ofE. ferox against fungal Strains (400 g).
InhibitionZ
oneDiameter(mm
)
Fungal Strains
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Fig. 10a: Aspergillus fumigates
Plate 10: Antifungal activity of methanolic seed and leaf extract of
E. ferox against Aspergillus fumigates and Cryptoccocus neoformans
SI=200 g, S2=400 g = Seed methanol extract. L1=200 g, L2=400 g = Leaf methanol
extract, Ab= Antibiotic; M=methanol
Fig. 10b: Cryptoccocus neoformans
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Fig. 11a: C. parapsilosis
Plate. 11: Antifungal activity of methanolic seed and leaf extract ofE. ferox against C.parapsilosis andRhizopus spp.
SI=200 g, S2=400 g = Seed methanol extract. L1=200 g, L2=400 g = Leaf methanol extract,
Ab= Antibiotic; M=methanol
Fig. 11b:Rhizopus spp
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Fig. 12a: C. albicans
Plate 12: Antifungal activity of methanolic seed and leaf extract ofE. ferox against C. albicans and
C. kruesie
SI=200 g, S2=400 g = Seed methanol extract. L1=200 g, L2=400 g = Leaf methanol extract,
Ab= Antibiotic; M=methanol
Fig. 12b: C. Kruesie
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Fig. 13a: P. notatum
Plate 13: Antifungal activity of
methanolic seed and leaf extract
ofE. ferox against P. notatum
C. stealoidea andC. tropicals .
SI=200g, S2=400 g = Seed methanol
extract.L1=200g,L2=400 g = Leaf
methanol extract, Ab= Antibiotic;
M=methanol
Fig. 13c: C. tropicals
Fig. 13b: C. stealoidea
Ph h i l C i f M h li E f
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Constituents tested for Leaf extract Seed extract
Alkaloids + +
Steroids _ +
Glycosides + +
Phenols + +
Tannins + +
Saponins _ +
Flavonoids + +
- = Not present , + = Present.
Phyto-chemical Constituents of Methanolic Extracts of
Euryale ferox
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A= Petiole methanol extractB= Seed methanol extract
C= Rhizome methanol extract
D= Leaf methanol extract
Solvent front =16.5cm
A B C D
Rf l f M h li f E l f
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Methanolic Extracts Rf. value
Seed 0.176, 0.41, 0.58, 0.63 and 0.84
Leaf 0.176, 0.35, 0.45 and 0.59
Petiole 0.26
Rhizome 0.11
Solvent front = 17 cm
solvents used = Chloroform: Methanol (7:3)
Rf values of Methanolic extracts of Euryale ferox
Extraction yield and macroscopic characteristics of the crude
t t f E l f
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Name of the
plant
Common
Name
Solvent Part used Macroscopic characteristics Extract/
yield (%)
Euryale ferox Jewer
Methanol
Chloroform
Petroleum ether
SeedSeed
Seed
Black crystalline powderBrown crystalline powder
Dark green shiny surface
3.84
3.07
0.3
Methanol
ChloroformPetroleum ether
Leaves
LeavesLeaves
Dark green shiny surface
Dark green powderYellowish green powder
9.44
1.740.6
Methanol
Chloroform
Petroleum ether
Rhizome
Rhizome
Rhizome
Dark black powder
Dark green shiny surface
A dark brown shiny surface
7.88
0.38
1.76
Methanol
Chloroform
Petroleum ether
Petiole
Petiole
Petiole
Dark green powder
Brown crystalline powder
Yellowish blue powder
10.9
0.98
1.66
extracts ofEuryale ferox
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CONCLUSION
O l th li t t f d d l f hibit d hi h t
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Only methanolic extracts of seed and leaf exhibited highest
antimicrobial activity.
Seed extracts exhibited higher inhibitory activity than leaf
extracts. Gram Positive strains were more susceptible than gram negative
strains.
Among the gram positive strains, S.aureusshows maximum zone of
inhibition i.e highest inhibitory activity. However, IZD of some gram negative strains ( i.e. P. aureoginosa)
were more than gram positive strains.
Among the fungal strains, Candida spp. and Pencillum notatum
were more susceptible to both extracts at higher
concentrations.
However C. neoformans,A. fumigates andRhizopus spp.
C.Parapsilosiswere completely resistant.
The results of MIC and MBC depicted that extracts ofE.
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p
ferox can be bactericidal in action and among
standard strains tested, S. aureus ATCC 25923 was
inhibited by least concentration of extracts used. Phyto-chemical screening of methanolic seed and leaf
extracts revealed the presence of alkaloids, phenols,
flavonoids, steroids and tannins while as steroids
and Saponins were present in only seed extract. The separation of methanolic extracts by thin layer
chromatography (TLC) yields different compounds
with seed extract showed five compounds of Rf
values 0.176, 0.41, 0.58, 0.63 and 0.84 while as
leaf extract yields four compounds with Rf. value of
0.176, 0.35, 0.45 and 0.59.
The substances that can inhibit pathogens and have
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p g
a little toxicity to host cells are considered good
for developing antimicrobial drugs.
Therefore, the antimicrobial activity ofE. feroxextracts on these UTI organisms reveals that it can
be safely used to treat these kinds of infections
after proper drug formulation.
The literature reveals that no prior work has been
done on evaluating antimicrobial activity ofE.
ferox and it is now expected after results that it
will be helpful in obtaining broad spectrum herbal
formulation as well as new antimicrobial
substances.
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