Background
Methods
Surface sterilization and initial culture establishment
Micropropagation
Extract preparation for antimicrobial screening
Collection and maintenance of test organisms
Antibiotics | UTI Pathogens | RTI Pathogens | |||||||
---|---|---|---|---|---|---|---|---|---|
Sample 5 (Escherichia coli) | Sample 9 (Escherichia coli) | Sample 11 (Escherichia coli) | Sample 19 (Klebsiella pneumoniae) | Sample 26 (Klebsiella pneumoniae) | Sample 31 (Klebsiella pneumoniae) | Sample 35 (Klebsiella pneumoniae) | Sample 36 (Klebsiella pneumoniae) | Sample 38 (Klebsiella pneumoniae) | |
Amoxiline | R | S | S | R | R | R | R | R | R |
Amikacin | S | S | S | R | R | S | S | R | R |
Aztreonam | R | R | R | R | R | NT | R | NT | NT |
Ceftriaxone | R | R | S | R | R | R | R | R | S |
Cefuroxime | R | R | S | R | I | R | R | R | R |
Ciprofloxacin | R | NT | I | I | I | R | R | R | R |
Piperacillin | S | S | S | R | S | S | NT | R | NT |
Cefoperazone | R | NT | S | R | R | S | S | R | R |
Ceftazidime | R | R | S | R | R | NT | NT | R | R |
Ofloxacin | R | R | R | I | S | NT | NT | NT | NT |
Sulbactum | S | R | S | R | S | R | R | S | NT |
Nitrofurantoin | S | R | S | NT | S | R | R | S | S |
Meropenum | S | NT | NT | I | R | S | R | R | R |
Doxycycline | S | R | S | S | S | NT | S | S | R |
Norfloxacin | R | S | S | I | S | R | R | R | R |
Antimicrobial screening by agar cup method
Minimum inhibitory concentration (MIC)
Minimum bactericidal and fungicidal concentration (MBC and MFC)
Statistical analysis
Results
Micropropagation
Concentration of plant growth regulators | Response (%) | Number of shoot buds induced per leaf explant (after 12 d of implantation) | No. of elongated (≥ 1 cm) shoot buds per leaf explant (after 42 d of implantation) | ||
---|---|---|---|---|---|
BAP (mg/L) | KIN (mg/L) | Spermidine (mM) | |||
– | – | – | 0 | 0a
| 0a
|
0.5 | – | – | 96.7 | 53.5 ± 1.7cd
| 11.7 ± 0.3c
|
1.0 | – | – | 100 | 76.7 ± 1.2de
| 16.1 ± 0.2d
|
1.5 | – | – | 100 | 92.4 ± 3.7ef
| 22.5 ± 0.4ef
|
2.0 | – | – | 100 | 84.1 ± 3.2e
| 19.2 ± 0.5e
|
2.5 | – | – | 100 | 78.2 ± 2.4e
| 13.8 ± 0.2cd
|
– | 0.5 | – | 90.0 | 45.0 ± 1.3bcd
| 9.5 ± 0.6c
|
– | 1.0 | – | 93.3 | 62.8 ± 1.9d
| 13.6 ± 0.7cd
|
– | 1.5 | – | 100 | 73.5 ± 2.2de
| 16.0 ± 0.3d
|
– | 2.0 | – | 100 | 81.3 ± 2.5e
| 18.2 ± 1.0de
|
– | 2.5 | – | 100 | 75.4 ± 1.9def
| 15.4 ± 0.8d
|
1.5 | – | 1.0 | 100 | 106.0 ± 3.2f
| 27.3 ± 0.7fg
|
1.5 | – | 1.5 | 100 | 114.2 ± 2.6fg
| 30.7 ± 0.9g
|
1.5 | – | 2.0 | 100 | 123.8 ± 3.4g
| 34.9 ± 1.2h
|
1.5 | – | 2.5 | 100 | 117.6 ± 3.1fg
| 31.4 ± 1.1gh
|
Antimicrobial activity
Solvent | Extract conc. (mg/cup) | Zone of Inhibition (mm) | |||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
MDR-UTI bacteria | MDR-RTI bacteria | MTCC bacteria | MTCC fungus | ||||||||||||
Sample 5 | Sample 9 | Sample 11 | Sample 19 | Sample 26 | Sample 31 | Sample 35 | Sample 36 | Sample 38 | MTCC 109 | MTCC 441 | MTCC 3160 | MTCC 870 | MTCC 1637 | ||
Acetone | 0.75 | – | 12 ± 0.11 | – | R | 12 ± 0.22 | – | 9 ± 0.28 | – | – | – | – | – | 8 ± 0.08 | – |
1.5 | 11 ± 0.11 | 13 ± 0.26 | – | R | 14 ± 0.32 | – | 11 ± 0.58 | 10 ± 0.56 | – | – | 8 ± 0.36 | 8 ± 0.33 | 10 ± 0.38 | 12 ± 0.22 | |
3.0 | 13 ± 0.24 | 15 ± 0.66 | – | R | 15 ± 0.36 | 9 ± 0.22 | 12 ± 0.11 | 11 ± 0.32 | 8 ± 0.11 | 12 ± 0.11 | 11 ± 0.32 | 12 ± 0.48 | 11 ± 0.28 | 14 ± 0.18 | |
4.5 | 15 ± 0.36 | 16 ± 0.33 | 11 ± 0.11 | R | 16 ± 0.14 | 11 ± 0.11 | 12 ± 0.56 | 13 ± 0.88 | 9 ± 0.28 | 14 ± 0.26 | 13 ± 0.44 | 14 ± 0.55 | 12 ± 0.56 | 16 ± 0.38 | |
6.0 | 15 ± 0.22 | 16 ± 0.48 | 13 ± 0.24 | R | 16 ± 0.18 | 11 ± 0.58 | 12 ± 0.22 | 13 ± 0.44 | 9 ± 0.22 | 14 ± 0.44 | 15 ± 0.48 | 14 ± 0.58 | 12 ± 0.88 | 19 ± 0.12 | |
Ethanol | 0.75 | – | 14 ± 0.18 | – | R | – | – | – | – | – | – | – | – | – | – |
1.5 | – | 16 ± 0.38 | – | R | – | 8 ± 0.28 | 9 ± 0.48 | – | – | – | – | – | – | 10 ± 0.26 | |
3.0 | – | 17 ± 0.22 | 12 ± 0.32 | R | – | 9 ± 0.44 | 10 ± 0.22 | – | 10 ± 0.21 | 11 ± 0.32 | 8 ± 0.38 | 10 ± 0.22 | 10 ± 0.21 | 14 ± 0.46 | |
4.5 | 11 ± 0.18 | 17 ± 0.18 | 14 ± 0.11 | R | – | 11 ± 0.22 | 11 ± 0.48 | 8 ± 0.32 | 11 ± 0.44 | 13 ± 0.22 | 10 ± 0.22 | 13 ± 0.33 | 11 ± 0.26 | 18 ± 0.55 | |
6.0 | 13 ± 0.22 | 17 ± 0.66 | 14 ± 0.62 | R | 10 ± 0.11 | 11 ± 0.22 | 12 ± 0.56 | 10 ± 0.44 | 11 ± 0.22 | 15 ± 0.48 | 12 ± 0.22 | 16 ± 0.48 | 11 ± 0.18 | 21 ± 0.38 | |
Methanol | 0.75 | 11 ± 0.26 | 13 ± 0.78 | – | R | 8 ± 0.33 | 7 ± 0.56 | – | – | 10 ± 0.28 | 11 ± 0.48 | – | 10 ± 0.22 | 12 ± 0.18 | 12 ± 0.18 |
1.5 | 12 ± 0.88 | 16 ± 0.18 | – | R | 10 ± 0.24 | 9 ± 0.88 | – | 9 ± 0.88 | 12 ± 0.32 | 13 ± 0.66 | – | 12 ± 0.48 | 14 ± 0.11 | 16 ± 0.77 | |
3.0 | 13 ± 0.48 | 18 ± 0.11 | – | R | 11 ± 0.18 | 11 ± 0.22 | 8 ± 0.11 | 11 ± 0.88 | 13 ± 0.12 | 13 ± 0.78 | 9 ± 0.48 | 14 ± 0.55 | 16 ± 0.22 | 18 ± 0.34 | |
4.5 | 14 ± 0.54 | 18 ± 0.66 | 10 ± 0.24 | R | 12 ± 0.38 | 12 ± 0.38 | 8 ± 0.88 | 12 ± 0.48 | 13 ± 0.21 | 13 ± 0.48 | 10 ± 0.52 | 14 ± 0.33 | 16 ± 0.38 | 18 ± 0.72 | |
6.0 | 14 ± 0.22 | 18 ± 0.68 | 13 ± 0.34 | R | 13 ± 0.62 | 13 ± 0.62 | 9 ± 0.32 | 12 ± 0.48 | 14 ± 0.22 | 13 ± 0.28 | 12 ± 0.66 | 14 ± 0.68 | 16 ± 0.18 | 18 ± 0.38 |
Microorganisms | Inhibition type | Inhibiting/cidal concentrations (mg/mL) of different solvent extracts | ||||
---|---|---|---|---|---|---|
Type | Bacteria / fungi | Strain | Acetone | Ethanol | Methanol | |
Pathogenic | MDR-UTI bacterial strains | Sample 5 | MIC | 5.0 | 7.5 | 5.0 |
MBC | 10.0 | 12.5 | 12.5 | |||
Sample 9 | MIC | 5.0 | 2.5 | 2.5 | ||
MBC | 10.0 | 5.0 | 5.0 | |||
Sample 11 | MIC | 5.0 | 5.0 | 7.5 | ||
MBC | 12.5 | 10.0 | 12.5 | |||
Sample 19 | MIC | R | R | R | ||
MBC | R | R | R | |||
Sample 26 | MIC | 2.5 | 25.0 | 10.0 | ||
MBC | 5.0 | 30.5 | 17.5 | |||
MDR-RTI bacterial strains | Sample 31 | MIC | 7.5 | 7.5 | 10.0 | |
MBC | 10.0 | 12.5 | 15.0 | |||
Sample 35 | MIC | 7.5 | 5.0 | 15.0 | ||
MBC | 12.5 | 7.5 | 17.5 | |||
Sample 36 | MIC | 5.0 | 10.0 | 7.5 | ||
MBC | 10.0 | 15.0 | 15.0 | |||
Sample 38 | MIC | 15.0 | 12.5 | 5.0 | ||
MBC | 17.5 | 17.5 | 7.5 | |||
Standard | MTCC bacterial strains | MTCC 109 | MIC | 5.0 | 5.0 | 7.5 |
MBC | 12.5 | 12.5 | 12.5 | |||
MTCC 441 | MIC | 7.5 | 12.5 | 10.0 | ||
MBC | 10.0 | 17.5 | 15.0 | |||
MTCC 3160 | MIC | 10.0 | 15.0 | 2.5 | ||
MBC | 12.5 | 22.5 | 7.5 | |||
MTCC fungal strains | MTCC 870 | MIC | 2.5 | 10.0 | 2.5 | |
MFC | 7.5 | 12.5 | 5.0 | |||
MTCC 1637 | MIC | 7.5 | 7.5 | 1.25 | ||
MFC | 12.5 | 12.5 | 5.0 |