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Department of Veterinary Pharmacology and Toxicology, Madras Veterinary College, Chennai 600 007
Department of Veterinary Pharmacology and Toxicology, Madras Veterinary College, Chennai 600 007
Department of Veterinary Pharmacology and Toxicology, Madras Veterinary College, Chennai 600 007
Ramesh
Aqueous
Aqueous
Terminalia chebula
Aqueous
Punica granatum Linn( pericarp)
Vitis vinifera
(seeds)
Alcoholic extract
Agar well diffusion assay -Hassawi and Kharma(2006) with slight modifications
Molten SDA
Cool
5 x106CFU/ml
Candida spp
Punch wells
Add the extract of
known
concentration to
each well
Incubate at 37ºC,24-48hrs
+ control - Amphotericin B
Measure the zone of inhibition -control- 5%DMSO
DIGITAL CALIPERS
Agar well diffusion assay for T.chebula
28
26
24
(Zone diameter mm)
22
20
18
16
14
12
10
8
6
4
2
0
20
Zone of inhibition(mm)
18
16
14
12
10
8
6
4
2
0
C12 801 R 783 800 845
Isolates
18
16
Zone of inhibition(m m )
14
12
10
Zone of 20
Isolate 15
200mg/ml 500mg/ml
10
14
12
Zone of
10
8
6
4
2
0
845 845
concentration of the extract(mg/ml )
300mg/ml
5%DMSO
300mg/ml
300mg/ml
150mg/ml
150mg/ml
500mg/ml
5%DMSO
150m
Grape seed extract –Isolate C12
500mg/ml 500mg/ml
lg/m
T.Chebula isolate 801
300mg/ml
500mg/ml
150mg/ml
T. chebula Isolate 801
300mg/ml
500mg/ml 150mg/ml
500mg/ml
Amphotericin B
10μg/ml
200mg/ml
Mangifera indica
Mangifera indica 500mg/ml
Isolate C12
Amphotericin B
10μg/ml Pomegranate
200mg/ml
200mg/ml
Isolate 845
Resazurin dye microbroth dilution method
(Sarkar et al.,2007)
5 x106CFU/ml
•100μl extract in 1st well
•50 μl sterile nutrient broth in 2-12th well
•Serial two fold dilution
96 WELL MICROTITRE PLATE
•10 μl dye and 30 μl sterile broth from 1-
12 wells
•Finally add 10 μl fungal
Highest dilution of suspension(5x106CFU/ml)
the extract •Loosely wrap and incubate at 37ºC for
showing inhibition 24-48hrs.
of fungal growth.
•Fungal growth-pink color
(The well before
Growth inhibition- blue color
pink color change
taken as MIC of
that extract
MIC Results
Sl.no Extract MIC value(mg/ml)
0.078mg/ml
0.156mg/ml
0.156mg/ml
0.625mg/ml
0.078mg/ml
0.625µg/ml
MIC results of T.chebula
0.3125mg/ml
0.3125mg/ml
0.625mg/ml
0.625mg/ml
0.625µg/ml
• All the extracts tested in this study posses
activity against yeast (candida spp) isolated from
bovine mastitis.
• Grape seed extract said to possess broad spectrum antibiotic
effect due to the presence of natural quaternary compound and
used in vaginal yeast infections in women as douche. (Dr.Leo
galland).
• T. chebula contains tannins, anthraquinone, chebulinic acid,
chebulegic acid ,shown to reduce growth of C. albicans.
(Elizabeth et al.,2005) .
• Gallotannins of Mangifera indica seed kernel shown to possess
activity against Bacillus subtilis (Ganzle and Schieber, 2010).
• Punica granatum Linn (Pomegranate) might be used as an
effective anti-bacterial agent against oral biofilm bacteria
(Pereira et al., 2006) they have shown to contain tannins.
• Antibacterial activity of guava leaves against mastitis causing
bacteria was studied by (Inchaisri et al.,2005).Guava leaf is rich
in flavanoids whose antibacterial activity was demonstrated by
several researchers (Abdelrahim et al.,2002).
Conclusion