Agar Disc Diffusion Lab Report

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.4 Agar Disk Diffusion
The antimicrobial tests were carried out according to disc diffusion tests (Lennette et al., 1985, Kim et al., 1995). Cells were streaked on MHA agar plate to obtain colonies at 37°C for 16 hrs after which they were resuspended in sterile saline solution to give an O.D of 0.1 at 600nm. On solidified 2% MHA plate, the bacterial culture is swabbed and sterile disc impression was made, 10 µl plant extracts was loaded on the agar plate. After 10 minutes the plate is incubated at 370C The diameter of the clear zone was measured.

Fig 9: Agar disk diffusion

5.5 Time Kill Assay
The basic concept of the Time-Kill Kinetic study is establishment of the rate at which a microorganism is killed by a product as a function of survival
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A34-a 384.55 DMSO
14. A49-a 336.38 DMSO 15. A41-b 366.38 DMSO
16. A05-b 322.36 DMSO
17. A37-a 386.17 DMSO
18. A39-a 322.13 DMSO
19. A45-a 336.15 DMSO
20. A35 303.31 DMSO
21. A38-a 312.39 DMSO
22. A38-b 312.39 DMSO
23. A51 330.26 DMSO
24. A03 254.71 DMSO
Table 10: List of synthetic compounds used in this study
Out of the above mentioned compounds; the following compounds were found to be active against given bacterial strains with MIC of:

Minimum inhibitory concentration(µM)

Compound S. aureus E. faecalis E. coli V. cholera H. influenzae L. monocytogens S. pyogenes
Rifampicin 3.90 1.95 31.25 0.485 >250 0.485 0.485
A15-b
>250 >250 >250 >250 >250 >250 250
A36-b
>250 >250 >250 >250 >250 >250 250
A37-a
125 125 125 125 125 125 125
A35
125 125 125 >250 >250 >250 >250
A38-b
250 >250 >250 >250 >250 >250 >250
A51
250 >250 >250 >250 >250 >250 >250
Table 11: List of synthetic compounds active against respective spp.

S.No. Acetone Extracts Chloroform Extracts Hexane Extracts Methanol Extracts Ethanol Extracts Water Extracts
1. 1 A 1 C 1 H 1 M 1 E 1 W
2. 2 A 2 C 2 H 2 M 2 E 2 W
3. 3 A 3 C 3 H 3 M 3 E 3 W
4. 4 A 4 C 4 H 4 M 4 E 4 W
5. 5 A 5 C 5 H 5 M 5 E 5 W
6. 6 A 6 C 6 H 6 M 6 E 6
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Extracts Inhibition zone (in cm) Strains
1. 5C 1.3 E.coli
2. 1A 1.5 E.coli
3. 3E 1.9 E.coli
4. 4E 2 E.coli
5. 6E 1.4 E.coli
6. 5E 1 E.coli
7. 1E 2 E.coli
8. 6M 1.9 E.coli
9. 6A 1.6 S.aureus
10. 2E 2 E.faecalis
11. 3E 1.9 E.faecalis
12. 4A 1.2

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