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added to the cups, aseptically and labeled, accordingly. reference standard (0.5 mg and 1 mg/ml conc.) were
The plates were kept undisturbed for at least 2 hrs at prepared by dissolving 5 mg and 10 mg of Griseofulvin
room temperature to allow diffusion of the solution in 10 ml of dimethylformamide to obtain a solutions of
properly, into nutrient agar medium. After incubation 50 μg/ml and 100 μg/ml concentration.
of the plates at 37±1°C for 24 hr. All the experiments
were carried out in triplicate. Simultaneously, controls The potato-dextrose-agar medium was sterilized by
were maintained employing 0.1 ml of autoclave at 121°C (15 lb/sq. inch), for 15 minutes.
dimethylformamide to observe the solvent effects. The Petri-plates, tubes and flasks plugged with cotton
were sterilized in autoclave at 121°C, for an hour. Into
Antifungal activity [19] each sterilized Petri-plate (10 cm diameter), about 30
Preparation of Potato dextrose agar media ml each of molten potato dextrose-agar medium
Potato - 100 gm inoculated with respective fungus (6 ml of inoculums
Dextrose - 10 g to 300 ml of potato-dextrose-agar medium) was
Distilled water- 500ml transferred, aseptically. After solidification of the
PH -5.6 medium at room temperature four cups of 6mm
All six essential oils were tested for their antifungal diameter were made in each plate with a sterile borer.
activity. The fungi employed for screening were Accurately 0.1 ml (100 μg/ml conc.) of test solution
Aspergillus flavus, Candida albicans, Microspora was transferred to the cups, aseptically and labeled,
gryseus and Aspergillus terus. The test organisms were accordingly. The reference standard 0.1 ml (50μg/ml
sub-cultured using potato dextrose agar medium. The conc., 100 μg/ml conc.) was also added to the cups in
tubes containing sterilized medium were inoculated each plate. The plates were kept undisturbed for at least
with test fungi and after 25°C for 48 hr. they were two hours at room temperature to allow diffusion of the
stored at 4° in a refrigerator. The inoculums was solution properly, into potato-dextrose-agar medium.
prepared by taking a loopful of stock culture to about Then the plates were incubated at 25°C for 48 hr. The
100 ml of nutrient broth; in 250 ml clean ad sterilized diameter of the zone of inhibition was read with help of
conical flasks. The flasks were incubated at 25°C for an ‘antibiotic zone reader’. The experiments were
24 hr. before use the solutions of test substances were performed in triplicate in order to minimize the errors.
prepared by a similar procedure described under the
Table-1: Anti-bacterial activity
Zone of inhibition (in mm)
Sl. No. Sample code
P. a. B.s. S. t. E. c.
a 8 5 2 2
A
1 b 10 3 1 3
c 0 4 4 3
a 0 4 4 1
2 N b 10 6 8 6
c 12 5 6 5
a 20 15 22 20
3 C b 22 18 17 22
c 23 16 15 23
a 30 23 21 25
4 E b 30 22 24 25
c 30 25 25 25
a 28 26 24 25
b 22 22 21 23
5 S
c 20 19 18 21
a 24 23 24 26
6 R b 22 22 20 24
c 21 20 19 22
S1 34 32 32 36
Standard
7 S2 31 31 30 33
Amoxicillin
S3 31 31 29 31
8 Control --- --- --- ---
A- Almondoil, N- Neem oil, C -Clove oil , E- Eucalyptus oil, S- Sandal wood oil, R-Rose oil
Sample Code
A-Almond oil, N- Neem oil, C -Clove oil , E- Eucalyptus oil, S- Sandal wood oil,
R- Rose oil
Name of the organisms
A. f- Aspergillus flavous, C. a- Candida albicans, M. g- Microspora gryseus, A. t- Aspergillus terus
Concentrations
a. 50% (1: 1) 5ml oil & 5ml DMF solvent (1%),
b. 50% (1: 1) 5ml oil (a) & 5ml DMF solvent (0.5%)
c. 50% (1: 1) 5ml oil (b) & 5ml DMF solvent (0.25%)