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Appendix I: Antibiotic Initial Solvent Final Diluent
Appendix I: Antibiotic Initial Solvent Final Diluent
Initial Solvent
Final Diluent
Cephalexin
B1
B1
Gentamicin
B2
B2
DMF
B4
Nystatin
(Dimethyl formamide)
Rifampicin
Methanol
B1
Streptomycin
Water
Water
Tetracycline
0.1N HCl
B3
Penicillin G
B1
B1
Kanamycin
Water
Water
B1
B2
K2HPO4
2.0g
K2HPO4
16.73g
KH2PO4
8.0g
KH2PO4
0.593g
Distilled Water
1.0 litre
Distilled Water
1.0 litre
pH
6.00.1
pH
8.00.1
B3
B4
KH2PO4
13.61g
K2HPO4
20.0g
Distilled Water
1.0 litre
KH2PO4
80.0g
pH
4.50.1
Distilled Water
1.0 litre
pH
6.00.1
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APPENDIX II
All media were sterilized at 1210C for 20 minutes unless otherwise stated.
Antibiotics & B-vitamins were sterilized by filtration.
1. Potassium tellurite agar medium:
Potassium tellurite
0.1g
Peptone
5.0g
Yeast extract
2.5g
Agar
20.0g
Distilled water
1.0 litre
pH
6.4
Soluble starch
10.0g
0.3g
KNO3
2.0g
MgSO47H2O
2.0g
K2HPO4
2.0g
MgSO47H2O
0.05g
CaCO3
0.02g
FeSO47H2O
0.01g
Agar
20.0g
Distilled water
1.0 litre
Nystatin
50.0mg
Benzyl Penicillin
0.8mg
164
14.0g
Agar
10.0g
Distilled Water
1.0 litre
pH
7 to 7.2
Nystatin
50.0mg
Benzyl penicillin
0.8mg
Bacto-Tryptone (Difco)
5.0g
3.0g
Distilled Water
1.0 litre
pH
7-7.2
Tripticase
15.0gm
Phytone
5.0gm
Sodium Chloride
5.0gm
Agar
15.0gm
Distilled Water
1.0 litre
4.0g
10.0g
Bacto-Dextrose (Difco)
4.0g
Distilled Water
1.0 litre
pH
7.3
Bacto-Agar
20.0g
165
20.0g
Agar
18.0g
1.0ml
Agar
18.0g
pH
7.2
1.0g
MgSO47H2O
1.0g
NaCl
1.0g
(NH4)2 SO4
2.0g
CaCO3
2.0g
Distilled water
500ml
1.0ml
pH
166
20.0g
1.0g
Glycerol
10.0g
1.0g
Distilled Water
1.0 litre
1.0ml
pH
7.0-7.4
Agar
20.0g
36.0g
1.0g
Distilled water
1.0 litre
pH
7.0 7.2
dehydrated (Difco)
15.0g
5.0g
0.5g
1.0 gm
Sodium thiosulphate
0.08g
Bacto-agar
15.0g
(K2HPO4)
167
0.64g
FeSO4 7H2O
0.11g
MnCl2 4H2O
0.79g
ZnSO4 7H2O
0.15g
Distilled water
100.0ml
C) Basal mineral salts agar (analytical reagent grade chemicals were used)
(NH4)2 SO4
2.64g
KH2PO4 (anhydrous)
2.38g
K2HPO4 3H2O
5.65g
MgSO4 7H2O
1.00g
1.0ml
Distilled water
1.0litre
pH
6.8 7.0
Agar (Difco)
15.0g
D) Complete medium
The sterile basal agar medium (C) was cooled to 600C and sterile carbon
source was aseptically added to give a concentration of approximately 1%.
168
5.0g
Beef extract
3.0g
Gelatin Powder
4.0g
Agar
15.0g
Distilled water
1.0litre
pH
7.0
1.0g
Agar
20.0g
100.0ml
Distilled water
1.0 litre
pH
7.6
Beef extract
3.0g
Starch soluble
2.0g
Agar
20.0g
Distilled water
1.0 litre
pH
7.6
169
3.0g
Peptone
5.0g
Potassium nitrate
1.0g
Distilled water
1.0 litre
pH
7.2
Meat extract
3.0gm
Peptone
5.0gm
Sodium chloride
5.0gm
Agar
15gm
Distilled water
1000ml
Yeast extract
1.0g
Beef extract
1.0g
Nz-amine A
2.0g
Agar
15.0g
Glucose
10.0g
Distilled water
1.0 litre
pH
7.3
200gm
Glucose (Dextrose)
20gm
Agar
20gm
Distilled water
1000ml
170
5.0gm
Peptone
5.0gm
Di-Potassium hydrogen
5.0gm
1.0 litre
Peptone
1.0gm
NaCl
5.0gm
K2HPO4
2.0gm
Phosphate
Distilled Water
21. Urea broth:
6ml
Agar
20.0
10ml
1000ml
0.2gm
1.0gm
1.0gm
Sodium Citrate
2.0gm
Sodium Chloride
5.0gm
Bromothmnol blue
0.08gm
Distilled water
1.0 litre
171
1.0gm
NaCl
5.0gm
Yeast extract
5.0gm
Glucose
10.0gm
Distilled water
1.0 litre
Dextrose
20gm
Peptone
10gm
Agar
15gm
Distilled water
1 litre
pH
5.6
172
APPENDIX III
Details of the test organisms employed
A) Gram-positive bacteria:
B. megaterium
B.subtilis
Staph.aureus
B) Gram-negative bacteria:
E.coli
Ps.fluorescens
P.vulgaris
K. pneumoniae
C) Fungi:
A.niger
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