International Journal of Universal Pharmacy and Life Sciences

International Journal of Universal Pharmacy and
Life Sciences
RESEARCH ARTICLE
ARTICALTICLE
Pharmaceutical Sciences
EFFECT OF MURVA (MAERUA OBLONGIFOLIA ) ON DIFFERENT
FUNGAL STRAINS - AN IN VITRO STUDY
Corresponding Author
ARULANANDRAJ.N *
Co-Authors
PUNITHAVANI.T1 AND INDUMATHY.S2
*- Department of Pharmaceutical Analysis, 1- Department of Pharmaceutics,
2- Department of Pharmacology,
Mother Theresa Post Graduate & Research Institute of Health Science, Gorimedu,
Puducherry- 605006.
ABSTRACT
Murva is an important ayurvedic drug used as one of the ingredients in many Ayurvedic preparations. Many
skin diseases such as tinea and ringworm caused by dermatophytes exist in tropical and semitropical areas.In
general, these fungi live in the dead, top layer of skin cells in moist areas of the body, such as between the toes,
the groin, and under the breasts. The aim of the present study was carried out to establish the antifungal activity
of aqueous and alcoholic extracts of Maerua oblongifolia. It was carried out by agar diffusion method. Serial
dilution was done to find out Minimum inhibitory Concentration (MIC) and disc diffusion method to carry out
the zone of inhibition (ZI). About 10mg of alcoholic & aqeuoes extracts of murva were weighed and dissolved
in Dimethyl sulphoxide (DMSO) and this solutions were used to carry out the In-vitro Antifungal studies. Seven
fungal organisms namely Aspergillus niger, Aspergillus flavus, Sacchromyces cereviaea, Penicillium notatum,
Trichoderma harzianum, cladosporium elatum and Bipolaris sorokiniana were used. The standards used were
Amphotericin B- 100units/disc from Himedia, Mumbai, Clotrimazole- 20μg/disc from Himedia, Mumbai.The
alcoholic and aqueous extracts of murva showed a significant antifungal activity at the test doses of different
concentration and the activity was very comparable to the standard drug Amphotericin and Clotrimazole.
KEY WORDS
163
Murva, Anti-fungal, Agar, Zone of inhibition, Minimum Inhibitory Concentration.
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INTRODUCTION
Medicinal plants represent a rich source of antimicrobial agents1. Many of the
plant materials used in traditional medicine are readily available in rural areas at
relatively cheaper than modern medicine 2. Plants generally produce many secondary
metabolites which constitute an important source of microbicides, pesticides and
many pharmaceutical drugs. Plant products still remain the principal source of
pharmaceutical agents used in traditional medicine 3. Murva is an important
ayurvedic drug used as one of the ingredients in many Ayurvedic preparations.
Maerua oblongifolia (Forsk.) A. Rich. (Capparaceae) is one of the botanical sources
of the Ayurvedic drug Murva. Ethanomedical survey reveals that Murva is used to
cure various diseases such as fever, stomach ache, skin infections, urinary calculii,
diabetes mellitus, epilepsy, pruritis, rigidity in lower limbs, and abdominal colic.In
India, the Ayurvedic systems of medicine has been existing for over three thousand
years, Charaka and Sushruta, two of the earliest Indian scholars had sufficient
knowledge of the properties of the Indian medicinal plants. The Vedas are the epic
poems, which contain rich material on the herbal medicine of that time 4. The
medicinal treatises like the Charakasamhita and Sushrutasamhita are esteemed even
to this day as the treasures of literature on indigenous medicine 5. Amongst the many
synonyms of this plant, one is ‘Dha-nurgunopayogya’ meaning ‘the plant whose
bark is being used for the bow-strings’. These synonyms have also contributes to the
existing confusion. Many skin diseases such as tinea and ringworm caused by
dermatophytes exist in tropical and semitropical areas.In general, these fungi live in
the dead, top layer of skin cells in moist areas of the body, such as between the toes,
the groin, and under the breasts. These fungal infections cause only a minor
irritation. Other types of fungal infections could be more serious. They can penetrate
into the cells and cause itching, swelling, blistering and scaling. In some cases,
fungal infections can cause reactions elsewhere in the body 6.
MATERIALS AND METHODS:
The aim of the present study was carried out to establish the antifungal
activity of aqueous and alcoholic extracts of Maerua oblongifolia. In the present
study, the antifungal activity was carried out by agar diffusion method. The
responses of the fungal microorganisms to the alcoholic and aqueous extracts were
recorded and compared with the standard reference drugs. The alcoholic and aqueous
extracts of Maerua oblongifolia, was used to determine the minimum inhibitory
concentration (MIC) and zone of inhibition (ZI). Serial dilution was done to find out
MIC and disc diffusion method to carry out the zone of inhibition. About 10mg of
alcoholic and aqueous extract was weighed and dissolved in 10ml of DMSO and
these solutions were used for the antifungal studies. The medium used for the study
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was agar medium and the method is agar diffusion method. Seven fungal organisms
namely Aspergillus niger, Aspergillus flavus, Sacchromyces cereviaea, Penicillium
notatum, Trichoderma harzianum, cladosporium elatum and Bipolaris sorokiniana
were procured from JIPMER, subcultured and used for the study. Sterilized
Petridishes, cork borer, beakers, glass syringes and test tubes were sterilized by dry
heat sterilization at 160º c for 1 hr in hot air oven were used for the study. The medis
used for the study were Sabouraud fluid media and Sabouraud’s Dextrose Agar
media from Himedia, Mumbai were used for the studies.Then serial dilution assay
was done using about 200ml of newly prepared sabouraud fluid media, sterilized and
distributed into sterile test tubes. One loopful of fungal suspension was inoculated
prior to distribution of media. One test tube served as control, 10 test tubes used for
ten different concentrations (100µg-1000µg) of alcohol extract and another test tubes
used similarly for aqueous extract. Then it is incubated. The agar medium was
prepared using about 100ml of sabouraud dextrose agar and autoclaved at 121˚ C for
15mins. It was distributed into 7 sterile beakers and seven different fungal strains
were inoculated into each beaker, and then poured into sterile petriplates. Media was
allowed to solidify then bores were made using sterile cork borer. Control, (DMSO)
alcohol and aqueous extract each 0.1ml was poured into the bores. Standard
antifungal discs were placed on the surface of agar. Entire procedure was carried out
in the laminar air flow bench. The petriplates were kept for 2h. For the drug to
diffuse into agar. The plates were incubated at 25 ˚ C for 48h. And the zone of
inhibition was measured. The standards used were Amphotericin B- 100units/disc
from Himedia, Mumbai ,Clotrimazole- 20μg/disc from Himedia, Mumbai.
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Table 1. Zone of inhibition of standard drugs, alcohol and aqueous extract of M.
oblongifolia
Alcohol extract
S. No
Fungal organisms
Cl
ZI
Amp -B
ZI
MIC
(µg)
ZI
Aqueous extract
MIC
(µg)
ZI
1.
Aspergillus niger
38 ±
1.414
44.5 ±
2.121
500
27.66 ±
1.856
550
11.33 ±
1.202
2.
Aspergillus flavus
24 ±
1.414
29.5 ±
0.7071
540
19.33 ±
0.8819
620
7.66 ±
0.8819
3.
Sacchromyces
cerevisae
14 ±
1.414
19.5 ±
0.7071
580
12.33 ±
0.8819
660
6.33 ±
2.33
4.
Cladospora carnonii
12 ±
1.414
8±
1.414
840
5.33 ±
1.453
860
1.666 ±
0.333
5.
Trichoderma
harzianum
45 ±
0.2828
50 ±
1.414
540
26.66 ±
1.453
660
13 ±
1.155
6.
Penicillum niger
27 ±
1.414
22 ±
1.414
860
13.33 ±
0.6667
900
7.66 ±
0.333
7.
Bipolaris spicefera
29 ±
2.828
25.5 ±
0.7071
860
12.66 ±
1.202
920
6.33 ±
0.8819
Cl- Clotrimazole; Amp-B – Amphotericin B;
ZI- Zone of Inhibition, Mean ± SEM; MIC- Minimum Inhibitory Concentratio
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1
2
3
4
5
6
7
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Volume-1/Issue-1/July-August 2011
RESULTS
Different concentrations of the test solutions inhibited the growth of the all
fungal strains with varying degree of sensitivity. The inhibitory zones were recorded
for the MIC concentration of alcoholic and aqueous extract of M. oblongifolia against
individual fungal organisms. All the values are mean of triplicates. The alcoholic
extract was more effective in the case of all the organisms compared to that of the
aqueous extract.
DISCUSSION
Plants provide abundant resources of antimicrobial compounds and have been
used for centuries to inhibit microbial growth. The increasing incidences of fungal
infections and gradual rise in resistance to available antibiotics had highlighted the
need to find more alternative antifungal agents from other sources. In the African
countries the genus Maerua is used as antifungal in traditional medicine. Antifungal
activity was done by agar diffusion method. Agar diffusion method was followed in
the antifungal studies of plants like Acorus calamus7Calotropis procera 8. Serial
dilution was done to find out minimum inhibitory concentration and agar disc
diffusion method was done to compare the extract with the standards Amphotericin–
B, Clotrimazole and the control. Both aqueous and alcohol extracts showed
significant antifungal activity on all the fungal organisms tested. But the effect shown
by alcohol extract was more compared to aqueous extract. Plants like Curtisia
dentate, Glaucium oxylobum and Epinetrum villosum showed the presence of
phytoconstituents such as triterpenoids and alkaloids might be responsible for
antifungal activity 9, 10 &11. Similarly the preliminary phytochemical constituents
studies showed the presence of triterpenoids and alkaloids could have responsible for
the activity. Since prehistoric times, people have used natural resources for medicinal
purposes. Folk medicines employ many plants to counteract diverse diseases
including skin infections. A Large number of plants have been reported by researchers
and practitioners of traditional medicine to be useful in the treatment of skin diseases.
Fungi that infect the skin, nails, and hair, generally called “ringworm” or “tinea” are
classified as dermatophytes. Agents that prevent growth of fungi are also derived
from natural sources. The results observed in the In-vitro antifungal activity of M.
oblongifolia were significant.
CONCLUSION
The In-vitro antifungal activity was carried out on the alcoholic and aqueous
extracts of the roots of Maerua oblongifolia showed very signifigant results.
Antifungal activity was done by agar diffusion method. Both the aqueous and the
alcoholic extract showed significant In-vitro antifungal activity with the various
corresponding MIC as per serial dilution assay respectively against different fungal
strains. The effect shown by alcoholic extract was more compared to aqueous
extract.Thus it was concluded that M. oblongifolia which is considered as one of the
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Volume-1/Issue-1/July-August 2011
botanical sources of the drug Murva can be used as a good substitute for antifungal
drug with further in vivo studies.
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