Antibacterial activity of naturally obtained plant that gives a antibacterial activity and some anti malarial activity functions also.This plant majorly seen in hills areas.

1. In the partial fulfillment of the requirements for the award of the
degree of
BACHELOR OF PHARMACY
By
P.V.SIVA KRISHNA (Y15PH2056)
Under The Guidance of
GOURI SANKAR.KANDUKURI M.pharm. (Ph.D)
Assistant Professor,
Dept.of pharmacognosy,
M.A.M.College of pharmacy
EVALUATION OF IN-VITRO ANTIBACTERIAL ACTIVITY OF
ACTINIOPTERIS RADIATA

2. CONTENTS
 Introduction
 Aim and Objectives
 Plant Profiles
 Materials and methods
 Results and Discussion
 Summary and conclusion
 Bibliography
2 M A M College of Pharmacy

3. INTRODUCTION
M A M College of Pharmacy3
The word “herb” has been derived from the Latin
word, “herba” and an old French word “herbe”. Now a days,
herb refers to any part of the plant like fruit, seed, stem,
bark, flower, leaf, stigma or a root, as well as a non-woody
plant. Earlier, the term “herb” was only applied to non-woody
plants, including those that come from trees and shrubs.
These medicinal plants are also used as food, flavonoid,
medicine or perfume and also in certain spiritual activities.

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 Herbal medicine is oldest healing system around the
world like Ayurveda and Unani medicines of India,
Chinese herbal medicine etc. The use of medicinal plants
in the treatment of diseases is becoming popular because
of synthetic drugs resistance and more side effects. In
recent decade different medicinal plants were reported
scientifically about their medicinal use and scientists are
isolating different biological active metabolites from them
and are became source for synthesis of new drugs in
treatment of diseases. In this point of view the present
work was carried out on to evaluate the antibacterial
activity of Actiniopteris radiata whole plant.

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Extraction techniques of Medicinal plants
Extraction, as the term is used pharmaceutically, involves
the separation of medicinally active portions of plant or
animal tissues from the inactive or inert components by
using selective solvents in standard extraction procedures.
The products so obtained from plants are relatively impure
liquids, semisolids or powders intended only for oral or
external use.These include classes of preparations known
as decoctions, infusions, fluid extracts, tinctures, pilular
(semisolid) extracts and powdered extracts. Such
preparations popularly have been called galenicals, named
after Galen, the second century Greek physician. The
purposes of standardized extraction procedures for crude
drugs are to attain the therapeutically desired portion and to
eliminate the inert material by treatment with a selective

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Methods of Extraction of Medicinal Plants
 Maceration
 Infusion
 Digestion
 Decoction
 Percolation
 Hot continuous extraction(Soxhlet)
 Aqueous alcoholic extraction by fermentation
 Counter current extraction
 Ultrasound extraction(Sonication)
 Supercritical fluid extraction

7. PLANT PROFILE
M A M College of Pharmacy7
Actiniopteris radiata
Taxonomic Hierarchy
Scientific classification:
Kingdom : Viridiplantae
Superkingdom : Eukaryota
Phylum : Embryophyta
Class : Filicopsida
Order : Polypodiales
Family : Pteridaceae
Genus : Actiniopteris
Species : A. radiata
Binomial Name : Actiniopteris
radiata

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Actiniopteris radiata (Actiniopteris = 'ray fern') is a fern
plants growing around the India belong to the family
Pteridaceae. The plants have posses’ different medicinal
values like antifertility, anti-tubercular but there is very less
scientific evidence (The Wealth of India, 2006). So, the
present work was carried out to identify antibacterial activity
of Actiniopteris radiata whole plant.
whole plant of Actiniopteris radiata

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Major chemical constituents:
 Hentriacontane
 Hentriacontanol
 β-sitosterol
 β-sitosterol palmitate
 β-sitosterol-D-glucoside
 Quercetin-3-rutinoside.
 Rutin
Geographical Location
 India, Burma, Sri Lanka, Afghanistan, Persia, Arabia,
Yemen, South Eastern Egypt, Tropical Africa, Australia
and Madagascar.

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Common names:
 Actiniopteris australis radiata
 Asplenium polydactylon Webb
 Asplenium radiatum
 Pteris radiata Bojer
 Acrostichum radiatum Poir.
 Actiniopteris australis sensu Sim
 Asplenium radiatum
 Acropteris radiata

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Vernacular names:
 Sanskrit: Mayursikha.
 Bombay: Mapursika.
 North-Western Provinces of India: Morpach.
 Kerala: Nanmukhappullu.
 Andhra Pradesh: Nemali adugu.
 English name: Peacock’s tail.
 kannada: Mayiladumsikha.
 Tamil: Mayiladumsikha.

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Medicinal importance:
 The plant is said to be anthelmintic, alterative and
astringent. It is used in the treatment of prolonged malarial
fevers and to arrest haemorrhages.
 The plant Actiniopteris is used as a styptic, anthelmitic,
dietary supplement in malnutrition, also used in acute
headache and in dysentery.
 Medicinal uses of this plant for relieving itching.
 The root of this plant promotes fertility.
 Actiniopteris used for control of blood pressure,
tuberculosis and cough.

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AIM AND OBJECTIVES
 AIM:
 To evaluate the antibacterial activity of Actiniopteris
radiate, Using soxhlated process with hexane, ethyl
acetate and hydro-alcoholic solvents successively.
 OBJECTIVES:
 The main objectives of the study are:
 To prepare the extract of Actiniopteris radiate using
suitable solvents.
 To study the anti-bacterial activity of extract of Actiniopteris
radiate solvent extract.
 To assess the anti-bacterial activity of solvent extract of
Actiniopteris radiate by using Zone of inhibition, MIC,
MBC, secondary metabolites analysis.

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PLAN OF WORK
Literature survey of Actiniopteris radiate
Collection of Actiniopteris radiate plant
Drying of Actiniopteris radiate in shade
Preparation of extract Actiniopteris radiate using suitable solvents
Secondary metabolites analysis, Zone of inhibition, MIC, MBC analysis of
extracts
Determine the antibacterial activity of Actiniopteris radiate using soxhlated process with
hexane, ethyl acetate and hydro-alcoholic solvents.

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MATERIALS AND METHODS
Plant material
The plant material Actiniopteris radiata was collected near
Guntur region, Andhra Pradesh, India and authenticated by
Dr. Prayaga Murthy. Pragada, Govt. Degree College,
Yeleswaram, E. Godavari, A.P. India.
Preparation of the extracts
The whole plant was air-dried in the shade and powdered.
100g of the fine air-dried powder was soxhlated with
solvents consecutively in the order of their increasing
polarity, with hexane (waxy oil), chloroform, ethyl acetate
and ethanol respectively, for more than 6 hrs with each
solvent; and the extracts were then concentrated in vacuo
to yield dense residues. The yields of the extracts were
12.5%, 8.9%, 5.0% and 6.5% for ethanol, ethyl acetate,
hexane and chloroform respectively. The samples were

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Chemicals:
The solvents and chemicals used in current research were
analytical grade and used standard drug ciprofloxacin was
procured from local market (make Dr. Reddy’s Laboratories).
Selected bacterial strains:
Gram positive and gram negative bacterial strains were
tested in the current In-vitro antibacterial activity of A. radiate
study (Table I). The bacterial strains were taken from
National Chemical Laboratory (NCL), Pune.

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S. NO Gram Positive Gram Negative
1.
Streptococus
pneumoniae
Pseudomonas
aeruginosa
2. Staphylococcus aureus K. pneumoniae
3. Clostridium sporogenes Escheichia coli
TABLE I: LIST OF BACTERIAL STRAINS

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QUALITATIVE ANALYSIS FOR CRUDE DRUGS
The extract prepared was tested for the type of chemical
constituents present by known qualitative tests.The
following tests were carried out on the extract to detect
various phyto constituents present in them.
QUALITATIVE ANALYSIS
 Test for flavonoids
 Test for glycosides
 Test for phenols
 Test for sterols
 Test for tannins

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Secondary Name of the test Result (+/-)
metabolites H C Ea E
Flavonoids Pews - + + +
NaOH - + +- +
Glycosides Keller-Kiliani + - + +
Conc.H2SO4 + - + +
Phenols Ellagic acid - - - +
Phenols - - - +
Saponins Foam + - - -
Sterols LibermanBuchard - + - -
Salkowski - - - -
Tannins Gelatin - - - +

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The plant extracts were also screened for qualitative
analysis to know the relative distribution of the secondary
metabolites which may be responsible for the potent
antibacterial activity. The chloroform extract of A. radiata
showed flavonoids and sterols while hexane extract
contained wax, glycosides and saponins. On the other hand,
ethylacetate extract contained flavonoids and glycosides,
and ethanol extract contained flavonoids, glycosides,
phenols, tannins and alkaloids (Table 2).
Determination of MIC and MBC
Minimum Inhibitory Concentration (MIC) and Minimum
Bacterial Concentration (MBC) were determined for the
extracts. The minimum concentration, at which there was no
visually detectable bacterial growth, was taken as MIC.
Extract concentration of 0.1mg to 7.5mg/ ml in steps of

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RESULTS AND DISCUSSION
The selected plant extracts were found to be posse's
dosage dependent antibacterial activity and as concentration
increased the activity was increased simultaneously on tested
bacterial strains. The results for hexane, ethyl acetate,
Chloroform and hydro-alcoholic extracts were showed in Tables
2, 3 and 4. . The extracts showed low activity at low
concentrations and no inhibition on bacterial growth.
The highest zone of inhibition was obtained with ethanol and
hexane extracts at a concentration of 25mg/ml against all the
tested bacterial strains. The MBC values correlated with MIC
values. Comparatively, MBC values were higher than the MIC
values. The lowest MIC values were observed in ethanol and
hexane extract, moderate MIC values were obtained in ethyl
acetate extract, and higher values were obtained in chloroform
extract. The ethanol extract showed least MIC and MBC values
against S. aureus (0.440 and 0.790 mg/ml) and highest MIC

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Zone of inhibitions of Actiniopteris radiata hexane
extract
Concentration of the extract (mg/100µL)
Ciprofloxacin
D
M
S
O
Name of the microorganism 4 8 15 25
(100µg/200µL) (100µL)
Zone of inhibition (in mm)
klebsiella pneumoniae 2.00 3.67±0.33 5.67±0.67 8.33±0.33 20.67±0.3 -
Staphylococcus aureus 1.33±0.33 3.33±0.33 6.33±0.33 8.67±0.33 19.67±0.33 -
Pseudomonas aeruginosa 1.33±0.33 2.67±0.33 4.33±0.33 8.00 21.33±0.33 -
Escheichia coli 1.67±0.33 4.00 7.00±0.58 9.33±0.88 24.33±0.67 -

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8.33
8.67
8
9.33
7
7.5
8
8.5
9
9.5
klebsiella pneumoniae Staphylococcus aureus Pseudomonas aeruginosa Escheichia coli
Zoneofinhibition(Inmm)
Name of the microorganism
Concentration of the extract (25mg/µL)
klebsiella pneumoniae Staphylococcus aureus Pseudomonas aeruginosa Escheichia coli

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SUMMARY AND CONCLUSION
 Studies were undertaken to test the extracts of A. radiata
against these pathogens. The highest activity was
observed in ethanol extract followed by hexane, ethyl
acetate and chloroform. When these extracts were
analyzed for secondary metabolites obtained results
were supporting the MIC and MBC findings.
 The chloroform extracts of A. radiata showed flavonoids
and sterols, while hexane extract contained wax, largely
glycosides and saponins. On the other hand,
ethylacetate extract contained both flavonoids and
glycosides. Ethanol extract of A. radiata contained
flavonoids, glycosides and phenols. The highest activity
of ethanol extract may be due to the presence of
flavonoids, glycosides and phenols. The secondary
metabolites of various chemical types present in A.

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 BIBLIOGRAPHY
 Reference

 WHO Global strategy for containment of antimicrobial resistance. www.who.int/eme-
documents/ antimicrobial_ resistance/docs/EGlobal_strat.pdf 2001.

 Archibald L, Phillips L, Monnet D, McGowan JE Jr, Tenover F, Gaynes R. Clin. Infect Dis
1997; 24: 211-215.

 Smith TL, Pearson ML, Wilcox KR, Cruz C, Lancaster MV, Robinson-Dunn B, Tenover
FC,Zervos MJ, Band JD, White E, Jarvis WR. New England J Med 1999; 340: 493-501.

 Shahidi Bonjar GH. J Med Sci 2004b; 4: 136-141.

 Cragg GM, Newman DJ and Snader KM. J Nat Prod 1997; 60: 52-60.

 Tally F. Drug Discovery Today 1999; 4: 395-398.

 Press JB. Monroe Wall Symposium Chemtracts- Organic chemistry 1996; 9: 286-298.

 Cox PA. In Ciba Found Symp 1994; 185: pp 25-36

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