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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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