Background
Methods
Collection and authentication of the plant material
Preparation of extracts
Crude extraction
Solvent fractionation
Phytochemical screening
Inoculum preparation and standardization
Antibacterial activity assay
Agar well diffusion
Determination of the Minimum Inhibitory Concentration (MIC)
Determination of Minimum Bactericidal Concentration (MBC)
Statistical analysis
Results
Antibacterial activity
Category of test | Concentration | Bacteria | |||||
---|---|---|---|---|---|---|---|
S. aureus
|
S. pyogen
|
S. pneumoniae
| |||||
Stand | Clinic | Stand | Clinic | Stand | Clinic | ||
Crude | 19.5 mg/well | 16.67 ± 0.88a3d2 | 16.67 ± 0.33a3c1d3 | 13.67 ± 0.88a3c2d3 | 12.33 ± 0.33a3c1d2 | 13.33 ± 0.33a3d2 | 11.67 ± 0.33a3c2d3 |
39 mg/well | 19.67 ± 0.88a3 | 19.00 ± 0.58a3d1 | 18.33 ± 0.67a3 | 16.33 ± 0.67a3 | 16.00 ± 1.00a2 | 15.33 ± 0.33a3d2 | |
78 mg/well | 22.00 ± 0.58a2 | 21.67 ± 0.33a2 | 20.33 ± 0.33a3 | 18.67 ± 0.33a3 | 19.33 ± 1.20a1 | 18.00 ± 0.00a3 | |
Cef 0.03 mg/disc* | 28.33 ± 0.58 | 26.33 ± 0.67 | NT | NT | NT | NT | |
Amp 0.01 mg/disc* | NT | NT | 27.33 ± 0.67 | 26.33 ± 1.20 | 24.00 ± 0.58 | 23.00 ± 0.58 | |
Methanol | 19.5 mg/well | 13.00 ± 0.58a3d2e2 | 11.67 ± 0.33 a3d2e3 | 12.67 ± 0.33a3d1 | 11.67 ± 0.33a3d2 | 12.33 ± 0.88a3d1 | 11.33 ± 0.33a3d3 |
39 mg/well | 14.33 ± 0.33a3f3 | 13.33 ± 0.33a3f3 | 14.00 ± 0.58a3f2 | 12.67 ± 0.33a3d1f3 | 14.33 ± 0.33a3 | 13.67 ± 0.33a3d1 | |
78 mg/well | 16.17 ± 0.44a3g3 | 15.33 ± 0.67a3g3 | 15.00 ± 0.58a3g3 | 15.00 ± 0.58a3g3 | 16.00 ± 0.58a3 | 15.33 ± 0.33a3g2 | |
Cef 0.03 mg/disc* | 27.67 ± 0.33 | 26.33 ± 0 .33 | NT | NT | NT | NT | |
Amp 0.01 mg/disc* | NT | NT | 28.00 ± 0.00 | 26.33 ± 0 .67 | 25.00 ± 0 .58 | 23.67 ± 0.33 | |
Chloroform | 19.5 mg/well | 11.67 ± 0.33a3d1e3 | 11.67 ± 0.33a3d1e3 | 12.00 ± 0.58 a3d2 | 10.67 ± 0.33a3d2 | 12.00 ± 0.58a3d2 | 11.00 ± 0.00a3d2 |
39 mg/well | 13.00 ± 0.58a3f3 | 13.33 ± 0.33a3f3 | 13.67 ± 0.33 a3f3 | 12.00 ± 0.58 a3f3 | 14.00 ± 0.58a3 | 13.00 ± 0.58a3f2 | |
78 mg/well | 14.33 ± 0.33a3g3 | 14.00 ± 0.58a3g3 | 15.00 ± 0.58 a3g3 | 13.67 ± 0.58a3g3 | 15.67 ± 0.33a3g1 | 15.00 ± 0.58a3g3 | |
Cef 0.03 mg/disc* | 27.00 ± 0.57 | 26.33 ± 0.33 | NT | NT | NT | NT | |
Amp 0.01 mg/disc* | NT | NT | 27.67 ± 0.33 | 27.00 ± 0.58 | 25.00 ± 0.58 | 23.67 ± 0.33 |
Category of test | Concentration | Bacteria | |||
---|---|---|---|---|---|
S. flexneri
|
E. coli
| ||||
Stand | Clinic | Stand | clinic | ||
Crude | 19.5 mg/well | 12.00 ± 0.58a3d2 | 11.67 ± 0.33 a3d2 | 11.67 ± .88a3d1 | 11.33 ± 0.67a3d2 |
39 mg/well | 14.00 ± 0.00a3d1 | 13.67 ± 0.33 a3d1 | 12.67 ± 0.33a3 | 12.67 ± 0.33a3 | |
78 mg/well | 17.00 ± 0.58a3 | 16.00 ± 0.58 a3 | 15.00 ± 0.00a3 | 14.50 ± 0.29 a3 | |
Cef 0.03 mg/disc* | NT | NT | 28.33 ± 0.88 | 27.00 ± 0.58 | |
Cip 0.005 mg/disc* | 26.33 ± 0.67 | 25.00 ± 0.58 | NT | NT | |
Methanol | 19.5 mg/well | 11.00 ± 0.00a3d2 | 10.67 ± 0.67a3d2 | 10.33 ± 0.33a3d3 | 9.00 ± 0.58a3c2d3 |
39 mg/well | 13.00 ± 0.58a3 | 12.67 ± 0.33a3 | 12.00 ± 0.58a3d1 | 11.67 ± 0.33a3d2 | |
78 mg/well | 15.00 ± 0.58a3 | 14.67 ± 0.33a3 | 14.50 ± 0.29a3 | 14.33 ± 0.33a3 | |
Cef 0.03 mg/disc* | NT | NT | 27.67 ± 0.33 | 26.67 ± 0.33 | |
Cip 0.005 mg/disc* | 27.00 ± 0.58 | 26.33 ± 0.67 | NT | NT | |
Chloroform | 19.5 mg/well | 10.00 ± 0.58a3d2 | 10.33 ± 0.33a3 | --- | --- |
39 mg/well | 11.33 ± 0.33a3f1 | 11.67 ± 0.67a3 | ---- | --- | |
78 mg/well | 13.33 ± 0.33 a3g2 | 12.33 ± 0.33a3g2 | --- | --- | |
Cip 0.005 mg/disc* | 25.00 ± 0.58 | 24.67 ± 0.33 | NT | NT |
Category of test | Concentration | Bacteria | ||
---|---|---|---|---|
P. aeruginosa
|
S. typhi
| |||
Stand | Clinic | stand | ||
Crude | 19.5 mg/well | 11.00 ± 0.58a3c1d2 | 10.67 ± 0.88a3 | 13.33 ± 0.33a3d2 |
39 mg/well | 13.33 ± 0.33a3 | 13.00 ± 0.58a3 | 14.00 ± 0.58a3d2 | |
78 mg/well | 14.33 ± 0.33 a3 | 14.00 ± 0.58a3 | 17.67 ± 0.67a3 | |
Cip 0.005 mg/disc* | 25.67 ± 0.33 | 24.33 ± 0.88 | 30.00 ± 0.58 | |
Methanol | 19.5 mg/well | 10.33 ± 0.33a3d1 | 10.33 ± 0.33a3c1d2 | 11.00 ± 0.58a3d2 |
39 mg/well | 12.00 ± 1.00a3 | 12.33 ± 0.33a3 | 12.67 ± 0.88a3d1 | |
78 mg/well | 13.67 ± 0.33a3 | 13.67 ± 0.33a3 | 15.67 ± 0.33a3 | |
Cip 0.005 mg/disc* | 24.33 ± 0.33 | 24.00 ± 0.58 | 31.00 ± 0.58 | |
Chloroform | 19.5 mg/well | --- | --- | 10.33 ± 0.33a3d2e1 |
39 mg/well | --- | --- | 11.67 ± 0.33a3 | |
78 mg/well | --- | --- | 13.00 ± 0.58a3g3 | |
Cip 0.005 mg/disc* | NT | NT | 30.33 ± 0.33 |
Minimum inhibitory concentration of crude extract and solvent fractions
Bacteria | Crude extract | Solvent fractions | ||
---|---|---|---|---|
Methanol fraction | Chloroform fraction | |||
MIC | MIC | MIC | ||
S. aureus
| Stand. | 2.03 ± 0.00 | 8.13 ± 0.00 | 8.13 ± 0.00 |
Clinc. | 2.03 ± 0.00 | 8.13 ± 0.00 | 16.25 ± 0.00 | |
S. pyogen
| Stand. | 4.06 ± 0.00 | 8.13 ± 0.00 | 8.13 ± 0.00 |
Clinc. | 4.06 ± 0.00 | 8.13 ± 0.00 | 16.25 ± 0.00 | |
S. pneumoniae
| Stand. | 8.13 ± 0.00 | 16.25 ± 0.00 | 16.25 ± 0.00 |
Clinc. | 8.13 ± 0.00 | 16.25 ± 0.00 | 16.25 ± 0.00 | |
S. flexneri
| Stand. | 4.06 ± 0.00 | 16.25 ± 0.00 | 16.25 ± 0.00 |
Clinc. | 5.42 ± 1.36 | 16.25 ± 0.00 | 16.25 ± 0.00 | |
E. coli
| Stand. | 8.13 ± 0.00 | 32.50 ± 0.00 | --- |
Clinc. | 8.13 ± 0.00 | 32.50 ± 0.00 | --- | |
P. aeruginosa
| Stand. | 8.13 ± 0.00 | 16.25 ± 0.00 | --- |
Clinic | 8.13 ± 0.00 | 16.25 ± 0.00 | --- | |
S. typhi
| Stand | 4.06 ± 0.00 | 16.25 ± 0.00 | 16.25 ± 0.00 |
Minimum bactericidal concentration of crude extract and solvent fractions
Bacteria | Crude extract | Solvent fractions | ||
---|---|---|---|---|
Methanol fraction | Chloroform fraction | |||
MBC | MBC | MBC | ||
S. aureus
| Stand. | 2.03 ± 0.00 | 16.25 ± 0.00 | 16.25 ± 0.00 |
Clinc. | 4.06 ± 0.00 | 16.25 ± 0.00 | 16.25 ± 0.00 | |
S. pyogen
| Stand. | 4.06 ± 0.00 | 13.54 ± 2.71 | 16.25 ± 0.00 |
Clinc. | 4.06 ± 0.00 | 16.25 ± 0.00 | 16.25 ± 0.00 | |
S. pneumoniae
| Stand. | 8.13 ± 0.00 | 32.50 ± 0.00 | 32.50 ± 0.00 |
Clinc. | 16.25 ± 0.00 | 32.50 ± 0.00 | 32.50 ± 0.00 | |
S. flexneri
| Stand. | 8.13 ± 0.00 | 32.50 ± 0.00 | 16.25 ± 0.00 |
Clinc. | 8.13 ± 0.00 | 32.50 ± 0.00 | 32.50 ± 0.00 | |
E. coli
| Stand. | 8.13 ± 0.00 | 32.50 ± 0.00 | --- |
Clinc. | 8.13 ± 0.00 | 65.00 ± 0.00 | --- | |
P. aeruginosa
| Stand. | 8.13 ± 0.00 | 32.50 ± 0.00 | --- |
Clinic. | 8.13 ± 0.00 | 32.50 ± 0.00 | --- | |
S. typhi
| Stand | 4.06 ± 0.00 | 16.25 ± 0.00 | 16.25 ± 0.00 |
Phytochemical constituents of the crude extract and solvent fractions
Metabolites tested | Crude extract | Solvent fractions | ||
---|---|---|---|---|
Chloroform fraction | Methanol fraction | Aqueous fraction | ||
Alkaloids | + | - | + | - |
Saponins | + | - | + | + |
Tannins | + | + | + | - |
Anthraquinones | + | + | - | - |
Polyphenols | + | + | + | - |
Terpenoids | + | + | + | - |
Flavonoids | + | + | + | - |
Cardiac glycosides | - | - | - | - |