amsa pachayappa
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S. Amsaveni, S. Pavulraj, D. Silambarasan, K. Saranya and S.Ramesh
Department of Veterinary Pharmacology and Toxicology,Madras Veterinary College, Chennai 600 007
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Mastitis is one of the most important diseases ofhigh yielding dairy animals.
Fungal mastitis -2-3% of clinical cases seen in veterinary
practice
Though there are potent antifungal drugs
they are not used in the treatment of fungal mastitis
Plants with antifungal properties can be exploited to
provide safe and cost effective treatment
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Plants selected in this study
Mangifera indica Linn (Mango seed
kernel)
Terminalia chebula
Psidium gujava Linn (guava leaves)
Punica granatum Linn( pericarp)
Vitis vinifera
(seeds)
Aqueous
Aqueous
Aqueous
Aqueous
Alcoholic extract
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0
2
4
6
8
10
1214
16
18
20
22
24
2628
(Zonedia
metermm)
C12 801 R 783 800 845Isolate numbers /concentration mg/ml
Agar well diffusion assay for T.chebula
150mg/ml 300mg/ml 500mg/ml
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0
24
68
1012
1416
1820Z
oneofin
hibition(mm)
C12 801 R 783 800 845
Isolates
Agar well diffusion assay for
Vitis vinifera seed extract
150mg/ml 300mg/ml 500mg/ml
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0
2
4
6
8
10
12
14
16
18
20
Zoneofinhibition(m
C12 R 800
Isolate
200mg/ml 500mg/ml
Agarwell diffusion assay for
Mango seedkernel extract
0
5
10
15
20
25
30
35
Zoneof
inhibition(mm)
R 800 801 845 783 C12
Isolate no
Agarwell diffusion assay forAmphotericin B
10/g/ml
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0
2
4
6
8
10
12
14
16
18
Zon
eof
inhibition(m
845 845
concentration of the extract(mg/ml)
Agar well diffusion assay forGuava leaf
extract and Pomegranate pericarp extract
10mg/ml 200mg/ml 10mg/ml 100mg/ml 200mg/ml
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Grape seed extract Isolate C12
300mg/ml
500mg/ml
5%DMSO
150mg/ml
500mg/ml
300mg/ml
Grape seed extract Isolate 800
5%DMSO
300mg/ml
150mg/ml
500mg/ml
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T.Chebula isolate 801
150mg/ml
300mg/ml
500mg/ml
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Mangifera indica
T. chebula Isolate 801
150mg/ml
300mg/ml
500mg/ml
200mg/ml
500mg/mlAmphotericin B
10g/ml
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Isolate C12
Mangifera indica
200mg/ml
500mg/ml
Amphotericin B
10g/ml
200mg/ml
Isolate 845
Pomegranate
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100l extract in 1st well
50l sterile nutrient broth in 2-12th well
Serial two fold dilution
10l dye and 30l sterile broth from 1-
12 wells
Finally add 10l fungal
suspension(5x106CFU/ml)
Loosely wrap and incubate at 37C for24-48hrs.
Fungal growth-pink color
Growth inhibition- blue color
5 x106CFU/ml
Highest dilution of
the extractshowing inhibition
of fungal growth.
(The well before
pink color change
taken as MIC of
that extract
96 WELLMICROTITRE PLATE
Resazurin dye microbroth dilution method
(Sarkaret al.,2007)
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MIC Results
Sl.no Extract MIC value(mg/ml)
1 Mangifera indica Linn (Mango
seed kernel)0.625
2 Psidium gujava Linn (guava
leaves)
2.5
3 Punica granatum Linn( pericarp) 5
4 Terminalia chebula 0.3125-0.625
5 Vitis vinifera (seed) extract
0.625-0.078Amphotericin B 10g/ml 0.625g/ml
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MIC grape seed extract
0.078mg/ml
0.156mg/ml
0.156mg/ml
0.625mg/ml
0.078mg/ml
0.625g/ml
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0.3125mg/ml
0.3125mg/ml
0.625mg/ml
0.625mg/ml
0.625g/ml
MIC results ofT.chebula
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All the extracts tested in this study possesactivity against yeast (candida spp) isolated frombovine mastitis.
Grape seed extract said to possess broad spectrum antibioticeffect due to the presence of natural quaternary compound andused in vaginal yeast infections in women as douche. (Dr.Leogalland).
T. chebula contains tannins, anthraquinone, chebulinic acid,chebulegic acid ,shown to reduce growth ofC. albicans.
(Elizabeth et al.,2005) . Gallotannins ofMangifera indica seed kernel shown to possess
activity against Bacillus subtilis (Ganzle and Schieber, 2010).
Punica granatum Linn (Pomegranate) might be used as aneffective anti-bacterial agent against oral biofilm bacteria(Pereira et al., 2006) they have shown to contain tannins.
Antibacterial activity of guava leaves against mastitis causingbacteria was studied by (Inchaisri et al.,2005).Guava leaf is richin flavanoids whose antibacterial activity was demonstrated byseveral researchers (Abdelrahim et al.,2002).
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Conclusion
This property can be exploited for
developing herbal formulations for mastitis
treatment.
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