658
Semi Defi ned Medium for Renibacterium salmoninarum
Amount/ 1 l Ingredient
Preparation
10 g
Tryptone
10 ml
Mineral salts
solution
Contains per 200 ml: EDTA dihydrate (disodium salt), 100 mg;
MgCl 2 , 4 g; CaCl 2 .2H 2 O, 1.4 g; FeCl 2 .6H 2 O, 100 mg;
ZnSO 4 .7H 2 O, 100 mg; MnSO 4 .4H 2 O, 100 mg; CuSO 4 .5H 2 O,
50 mg; CoCl 2 .6H 2 O, 50 mg; Na 2 MoO 4 .2H 2 O, 50 mg
10 ml
Nitrogen
compounds
Contains per 100 ml: uracil, 50 mg; guanine, 50 mg; adenine,
50 mg; xanthine, 50 mg
1 mg each
Vitamins
Nicotinic acid, ribofl avin, thiamine, calcium pantothenate
2 mg each
Vitamins
Pyridoxal HCl, pyridoxine HCl (prepared after Rogosa et al .
1961 )
20 ml
Buffer (pH 6.8) Contains per 100 ml: K 2 HPO 4 , 15 g; KH 2 PO 4 , 15 g; sterilised
at 121 °C/15 min
8 ml
Cysteine HCl
Prepared immediately prior to addition as a solution (12.5 %
[w/v]) in N NaOH to give a pH of 6.8; fi lter sterilised
2 ml
Glucose
Prepared as a solution (50 % [w/v]) in distilled water; sterilised
at 115 °C/20 min
Note: tryptone, mineral salts solution and nitrogen compounds are added to distilled water, the pH
adjusted to 6.8 with N NaOH, and the medium dispensed in 190 ml amounts in 800 ml capacity
Erlenmeyer fl asks. Sterilisation is a 121 °C/15 min. After cooling to ~50 °C, the pre-sterilised buffer, glucose, cysteine HCl and vitamin solutions are added to a fi nal volume of 200 ml. Agar to 1 %
(w/v) may be incorporated, as necessary.
Shotts and Waltman Medium for the Isolation of Edwardsiella
ictaluri (Shotts and Waltman 1990 )
1 % (w/v) tryptone
1 % (w/v) yeast extract
0.125 % (w/v) phenylalanine
0.12 % (w/v) ferric ammonium citrate
0.0003 % (w/v) bromothymol blue
0.1 % (w/v) bile salts
1.5 % (w/v) agar
980 ml of distilled water
dissolve by boiling, cool to 50 °C and adjust pH to 7.0, sterilise at 121 °C for 15 min,
cool to 50 °C and add mannitol (fi lter sterilised) to 0.35 % (v/v) and colistin sulphate to 10 μg/ml.
12 Isolation/Detection
Semi Defi ned Medium for Renibacterium salmoninarum
Amount/ 1 l Ingredient
Preparation
10 g
Tryptone
10 ml
Mineral salts
solution
Contains per 200 ml: EDTA dihydrate (disodium salt), 100 mg;
MgCl 2 , 4 g; CaCl 2 .2H 2 O, 1.4 g; FeCl 2 .6H 2 O, 100 mg;
ZnSO 4 .7H 2 O, 100 mg; MnSO 4 .4H 2 O, 100 mg; CuSO 4 .5H 2 O,
50 mg; CoCl 2 .6H 2 O, 50 mg; Na 2 MoO 4 .2H 2 O, 50 mg
10 ml
Nitrogen
compounds
Contains per 100 ml: uracil, 50 mg; guanine, 50 mg; adenine,
50 mg; xanthine, 50 mg
1 mg each
Vitamins
Nicotinic acid, ribofl avin, thiamine, calcium pantothenate
2 mg each
Vitamins
Pyridoxal HCl, pyridoxine HCl (prepared after Rogosa et al .
1961 )
20 ml
Buffer (pH 6.8) Contains per 100 ml: K 2 HPO 4 , 15 g; KH 2 PO 4 , 15 g; sterilised
at 121 °C/15 min
8 ml
Cysteine HCl
Prepared immediately prior to addition as a solution (12.5 %
[w/v]) in N NaOH to give a pH of 6.8; fi lter sterilised
2 ml
Glucose
Prepared as a solution (50 % [w/v]) in distilled water; sterilised
at 115 °C/20 min
Note: tryptone, mineral salts solution and nitrogen compounds are added to distilled water, the pH
adjusted to 6.8 with N NaOH, and the medium dispensed in 190 ml amounts in 800 ml capacity
Erlenmeyer fl asks. Sterilisation is a 121 °C/15 min. After cooling to ~50 °C, the pre-sterilised buffer, glucose, cysteine HCl and vitamin solutions are added to a fi nal volume of 200 ml. Agar to 1 %
(w/v) may be incorporated, as necessary.
Shotts and Waltman Medium for the Isolation of Edwardsiella
ictaluri (Shotts and Waltman 1990 )
1 % (w/v) tryptone
1 % (w/v) yeast extract
0.125 % (w/v) phenylalanine
0.12 % (w/v) ferric ammonium citrate
0.0003 % (w/v) bromothymol blue
0.1 % (w/v) bile salts
1.5 % (w/v) agar
980 ml of distilled water
dissolve by boiling, cool to 50 °C and adjust pH to 7.0, sterilise at 121 °C for 15 min,
cool to 50 °C and add mannitol (fi lter sterilised) to 0.35 % (v/v) and colistin sulphate to 10 μg/ml.
12 Isolation/Detection