Preparation of yeast water: suspend 200.0 g of pressed yeast in 1000.0 ml of tap water and twice filter hot through a paper filter or centrifuge.

Boil 200 g scrubbed and sliced potatoes in 1000.0 ml water for 1 hour, filter cold through a cotton-gauze filter, fill up distilled water to 1000.0 ml, add agar. Do not use new potatoes.

Cook 20 g of oatmeal in 1000.0 ml tap water for 20 min, filter through 2 layers of gauze, dilute to 1000.0 ml and add 15.0 g agar.

Boil 200 g scrubbed and sliced potatoes in 1000.0 ml water for 1 hour, filter cold through a cotton-gauze filter, add water to the initial volume, add glucose and agar. Do not use new potatoes.

Boil potato and carrot in 1000.0 ml of water for 1 hour, filter cold through a cotton-gauze filter, add water to the initial volume and add agar. Do not use new potatoes.

Preparation of yeast water: suspend 200.0 g of pressed yeast in 1000.0 ml of tap water and twice filter hot through a paper filter or centrifuge.

Boil manure in 1000.0 ml of water for 10 min, then keep for 16-20 hours, filter through 1-2 layers of filter paper, adjust to the initial volume, add agar.

Boil peas in 1000.0 ml of water, filter through gauze, add water to the initial volume; add phosphate, sucrose and agar.

Sterilize glucose and mannitol separately (in 50.0 ml H₂O) and add to the medium after autoclaving.

Boil 50.0 g of cabbage in 1000.0 ml of water, filter cabbage, adjust the volume of broth to the initial value.

Pour 9000.0 ml of distilled water into 3.0 kg of curd and add 150.0 of dry Aspergillus terricola mycelium, shake, pour chloroform, seal with a stopper. Decoct at 37 °C for 10 days, adjust pH to 7.0 with 1 N NaOH. The decoction shall contain 400 mg% amine nitrogen and 300 mg% tryptophan.

10 ml filter-sterilized 20% urea solution is added aseptically after autoclaving to 1000 ml cooled, molten, agar. The medium is then immediately dispensed aseptically.

Dry garden soil, rich in organic material, in the air by spreading in a thin layer, comminuting and stirring.Then sieve through a rough sieve, and mix 400 g of soil with 960 ml of tap water. Autoclave at 121°C for 1 hour at the end of the day but leave the autoclave open overnight. Filter the cooled extract through filter paper, autoclave 300 ml of filtrate at 121°C for 20 min and allow to stay for 2 weeks or longer to settle the sediment. Decant the clear supernatant liquid and use to prepare the medium.

Autoclave solutions 1 and 2 separately at 121°C for 15 min, solution 3 at 111°C for 30 min and mix aseptically.

pour 9000.0 ml of distilled water into 3.0 kg of curd andadd 150.0 of dry Aspergillus terricola mycelium, shake, pour chloroform, seal with a stopper. Decoct at 37°C for 10 days, adjust pH to 7.0 with 1N NaOH. The decoction shall contain 400 mg% amine nitrogen and 300 mg% tryptophan.

mix cow manure (fresh) 1.0 kg and distilled water 3000.0 ml. Boil, squeeze through gauze into a bottle and dilute to 3000.0 ml.

Preparation of soil extract: dry garden soil, rich in organic material, in the air by spreading in a thin layer, comminuting and stirring. Then sieve through a rough sieve, and mix 400 g of soil with 960 ml of tap water. Autoclave at 121°C for 1 hour at the end of the day but leave the autoclave open over night. Filter the cooled extract through filter paper, autoclave 300 ml of filtrate at 121°C for 20 min and allow to stay for 2 weeks or longer to settle the sediment. Decant the clear supernatant liquid and use to prepare the medium.

Autoclave at 111°C for 20 min. Vitamin B₁₂ sterilize separately by filtration.

Boil beans in 1000.0 ml of water, filter through gauze, add water to the initial volume; add phosphate, sucrose and agar.

First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Final pH 7.0 (with KOH).

Boil 200 g scrubbed and sliced potatoes in 1000.0 ml water for 1 hour, filter cold through a cotton-gauze filter, add water to the initial volume, add glucose and agar. Do not use new potatoes.

Autoclave phosphate and salt solutions at 121°C for 15 min, base medium at 111°C for 30 min.

Autoclave solutions 1 and 3 separately under N₂ at 121°C for 15 min, add 5.0 ml of the filter sterilized vitamin solution to 1000.0 ml of sterile solution 1. Solution 2 (autoclave at 121°C for 15 min) is not to be kept for long.

91. THERMODESULFOBACTERIUM MEDIUM

Solution 1:

Na₂SO₄ 3.0 g

NH₄Cl 1.0 g

MgCl₂ x 6 H₂O 0.2 g

KH₂PO₄ 0.3 g

Na₂HPO₄ x 12 H₂O 2.0 g

FeSO₄ x 7 H₂O 1.5 mg

Resazurin 1.0 mg

Distilled water 930.0 ml

Solution 2:

Trace element solution (see below) 10.0 ml

Solution 3:

Yeast extract 1.0 g

Distilled water 25.0 ml

Solution 4:

Na-lactate 4.0 g

Distilled water 25.0 ml

Solution 5:

Na₂S x 9 H₂O 0.5 g

Distilled water 6.0 ml

Solution 6:

Vitamin solution (see below) 5.0 ml

Trace element solution:

Nitrilotriacetic acid 12.8 g

FeCl₂ x 4 H₂O 0.2 g

MnCl₂ x 4 H₂O 0.1 g

CoCl₂ x 6 H₂O 0.17 g

CaCl₂ x 2 H₂O 0.1 g

ZnCl₂ 0.1 g

CuCl₂ 0.02 g

Na₂MoO₄ x 2 H₂O 0.01 g

NiCl₂ x 6 H₂O 0.026 g

NaCl 1.0 g

Na₂SeO₃ x 5 H₂O 0.02 g

Distilled water 1000.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

Ca DL-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

pH 6.8-7.0

Solutions 1, 3, 4 and 5 autoclave under N₂ at 121°C for 15 min.

Solution 6 is filter sterilized. Before use, neutralize solution 5 by dropwise of 1 N HCl.

92. DESULFOVIBRIO MEDIUM WITH LACTATE

Solution 1:

K₂HPO₄ 0.5 g

NH₄Cl 1.0 g

CaCl₂ , 6 H₂O 0.1 g

MgSO₄ , 7 H₂O 2.0 g

Na₂SO₄ 1.0 g

Na-lactate 5.0 g

Yeast extract 1.0 g

Resazurin 0.01 g

Cysteine 0.5 g

Distilled water 950.0 ml

Solution 2:

NaHCO₃ 4.0 g

Distilled water 40.0 ml

Solution 3:

Na₂S , 9 H₂O 300.0 mg

Distilled water 6.0 ml

Solution 4:

FeSO₄ , 7 H₂O 0.4 g

Distilled water 10.0 ml

pH 6.8

Autoclave all solutions separately at 121°C for 15 min. Solution 1 bring to boil while simultaneously bubbling a mixture of oxygen-free gas composed of 97% N₂ and 3% H₂ through the mixture and sterilize in atmosphere of this gas mixture. Solution 3 sterilize in atmosphere of N_2.

93. AZOSPIRILLUM BRASILIENSE MEDIUM 1

Ca-malate 10.0 g

K₂HPO₄ 1.0 g

MgSO₄ x 7 H₂O 0.5 g

CaCl₂ x 2 H₂O 0.02 g

Distilled water 1000.0 ml

pH 6.5

Autoclave at 121°C for 15 min.

94. MEDIUM FOR CARBON MONOOXIDE OXIDIZERS

Na₂HPO₄ x 12 H₂O 4.5 g

KH₂PO₄ 0.75 g

NH₄Cl 1.5 g

MgSO₄ x 7 H₂O 0.2 g

CaCl₂ x 2 H₂O 0.03 g

Fe(NH₄)-citrate 0.018 g

Agar (if necessary) 1.2 g

Trace element solution SL-6 (see below) 1.0 ml

Vitamin solution (see below) 10.0 ml

Distilled water 1000.0 ml

Trace element solution SL-6:

ZnSO₄ x 7 H₂O 0.1 g

MnCl₂ x 4 H₂O 0.03 g

H₃BO₃ 0.3 g

CoCl₂ x 6 H₂O 0.2 g

CuCl₂ x 2 H₂O 0.01 g

NiCl₂ x 6 H₂O 0.02 g

Na₂MoO₄ 0.03 g

Distilled water 1000.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl x 2 H₂O 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

D-Ca-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

pH 7.0

Autoclave at 121°C for 15 min, add vitamin solution sterilized by filtration. For chemoautotrophic growth incubate under gas atmosphere of a) 20-80% CO₂ + 10% O₂ + 0-70% N₂ or b) 70% H₂ + 20% O₂ + 10% CO₂ adding 2.5 g NaHCO₃ per liter of medium. For chemoorganotrophic growth add 3.0 g sodium acetate and incubate under air.

95. DESULFOTOMACULUM ACETOXIDANS MEDIUM

Solution 1:

NaCl 1.17 g

MgCl₂ , 6 H₂O 0.4 g

KCl 0.3 g

CaCl₂ , 2 H₂O 0.15 g

NH₄Cl 0.27 g

KH₂PO₄ 0.2 g

Na₂SO₄ 2.84 g

Na-acetate 1.4 g

Na-butyrate 1.4 g

Yeast extract 1.0 g

Vitamin solution (see below) 1.0 ml

Trace element solution 1.0 ml (see below)

Distilled water 940.0 ml

Solution 2:

Na₂S , 9 H₂O 0.36 g

Distilled water 10.0 ml

Sodium bicarbonate for alkalization:

NaHCO₃ 4.5 g

Distilled water 50.0 g

Vitamin solution:

p-Aminobenzoic acid 4.0 mg

D(+)-Biotin 1.0 mg

Thiamine-HCl 10.0 mg

Distilled water 100.0 ml

Trace element solution:

FeCl₂ , 4 H₂O 1.5 g

ZnCl₂ 68.0 mg

MnCl₂ , 4 H₂O 100.0 ml

H₃BO₃ 62.0 mg

CoCl₂ , 6 H₂O 120.0 ml

Na₂MoO₄ , 2 H₂O 24.0 mg

HCl (0.05 M) 1000.0 ml

pH 7.0-7.2

Solution 1 is boiled before sterilization for a few minutes being flushed with gas mixture of 80% N₂ and 20% CO₂ with sodium bicarbonate added until an equilibrium pH of 6.9 - 7.1. Solution 1 is autoclaved under this gas mixture at 121°C for 15 min. Solution 2 is autoclaved under 100% N₂ at 121°C for 15 min. Vitamin solution is filter sterilized.

96. DESULFONEMA LIMICOLA MEDIUM

Solution 1:

Na₂SO₄ 3.0 g

NaCl 13.0 g

MgCl₂ , 6 H₂O 2.2 g

CaCl₂ , 2 H₂O 0.15 g

KCl 0.5 g

KH₂PO₄ 0.2 g

NH₄Cl 0.3 g

Distilled water 850.0 ml

Solution 2:

Trace element solution SL-10 (see below) 1.0 ml

Solution 3:

NaHCO₃ 5.0 g

Distilled water 10.0 ml

Solution 4:

Na-acetate , 3 H₂O 2.5 g

Distilled water 100.0 ml

Solution 5:

Disodium succinate 0.1 g

Distilled water 1.0 ml

Solution 6:

Vitamin solution (see below) 5.0 ml

Solution 7:

AlCl₃ , 6 H₂O 245.0 mg

Distilled water 5.0 ml

Solution 8:

Na₂CO₃ 170.0 mg

Distilled water 1.6 ml

Solution 9:

Rumen fluid, clarified 20.0 ml

Solution 10:

Na₂S , 9 H₂O 0.4 mg

Distilled water 10.0 ml

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ , 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ , 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ , 6 H₂O 190.0 mg

CuCl₂ , 2 H₂O 2.0 mg

NiCl₂ , 6 H₂O 24.0 mg

Na₂MoO₄ , 2 H₂O 36.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

Ca DL-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

pH 7.6

The trace element solution preparation: FeCl₂ , 4 H₂O is dissolved firstly in HCl, and then is mixed with water and other salts are dissolved in the sequence indicated. Solution 1 is boiled for a few minutes, cooled to room temperature, gassed with 80% N₂ + 20% CO₂ gas mixture to reach a pH of around 6, then autoclaved anaerobically under the same gas. Solutions 2, 3, 5, 9, and 10 are autoclaved separately under nitrogen at 121°C for 15 min, solution 3 is filter-sterilized and flushed with 80% N₂ + 20% CO₂ to remove dissolved oxygen. Solution 6 is filter-sterilized and outgassed with N₂. Solutions 7 and 8 are combined before autoclaving at 121°C for 15 min. Solutions 2 to 10 are added to the sterile cooled solution A in the sequence as indicated. The completed medium is distributed anaerobically under 80% N₂ + 20% CO₂ into appropriate vessels.

97. DESULFONEMA MAGNUM MEDIUM

Solution 1:

Na₂SO₄ 3.0 g

NaCl 21.0 g

MgCl₂ , 6 H₂O 5.5 g

CaCl₂ , 2 H₂O 1.35 g

KCl 0.5 g

KH₂PO₄ 0.2 g

NH₄Cl 0.3 g

Distilled water 850 ml

Solution 2:

Trace element solution (see below) 1.0 ml

Solution 3:

NaHCO₃ 2.5 g/l

Distilled water 100.0 ml

Solution 4:

Na-benzoate 0.6 g

Distilled water 10.0 ml

Solution 5:

Disodium succinate 0.1 g

Distilled water 1.0 ml

Solution 6:

Vitamin solution (see below) 5.0 ml

Solution 7:

AlCl₃ x 6H₂O 245.0 mg

Distilled water 5.0 ml

Solution 8:

Na₂CO₃ 170.0 mg

Distilled water 1.6 ml

Solution 9:

Rumen fluid, clarified 20.0 ml

Solution 10:

Na₂S , 9 H₂O 0.4 mg

Distilled water 10.0 ml

Trace element solution:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ , 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ , 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ , 6 H₂O 190.0 mg

CuCl₂ , 2 H₂O 2.0 mg

NiCl₂ , 6 H₂O 24.0 mg

Na₂MoO₄ , 2 H₂O 36.0 mg

Na₂SeO₃ 3.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

Ca DL-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

pH 7.6

The trace element solution preparation: FeCl₃ x 4H₂O is dissolved firstly in HCl, then is mixed with water and other salts are dissolved in the sequence indicated. Solution 1 is boiled for a few minutes, cooled to room temperature, gassed with 80% N₂ + 20% CO₂ gas mixture to reach a pH of around 6, then autoclaved anaerobically under the same gas. Solutions 2, 3, 5, 9, and 10 are autoclaved separately under nitrogen at 121°C for 15 min, solution 3 is filter-sterilized and flushed with 80% N₂ + 20% CO₂ to remove dissolved oxygen. Solution 6 is filter-sterilized and outgassed with N2. Solutions 7 and 8 are combined before autoclaving at 121°C for 15 min. Solutions 2 to 10 are added to the sterile cooled solution A in the sequence as indicated. The completed medium is distributed anaerobically under 80% N₂ + 20% CO₂ into appropriate vessels.

98. WORT AGAR 7 B WITH 2% CaCO₃

Malt extract Balling 7 degrees 1000.0 ml

CaCO₃ 20.0 g

Agar 20.0 g

Autoclave at 111°C for 30 min.

99. MEDIUM YE

Yeast extract 30.0 g

Ethanol 20.0 ml

Agar 20.0 g

Distilled water 1000.0 ml

pH 5.0-6.0

Autoclave at 121°C for 15 min without ethanol.

Apply filter sterilized ethanol (0.1 ml/test tube) onto the surface of agar slants.

100. SAP-2 AGAR

Tryptone 1.0 g

Yeast extract 1.0 g

Agar 20.0 g

Sea water 1000.0 g

pH 7.4

Sterilize at 121°C for 15 min.

101. SOIL AGAR

Yeast extract 2.0 g

Tryptone 1.0 g

Na-acetate 1.0 g

Soil extract 50.0 ml

Agar 20.0 g

Distilled water add to 1000.0 ml

pH 7.4

Sterilize at 121°C for 15 min.

Preparation of soil extract: dry garden soil, rich in organic material, in the air by spreading in a thin layer, comminuting and stirring. Then sieve through a rough sieve, and mix 400 g of soil with 960 ml of tap water. Autoclave at 121°C for 1 hour at the end of the day but leave the autoclave open overnight. Filter the cooled extract through filter paper, autoclave 300 ml of filtrate at 121°C for 20 min and allow to stay for 2 weeks or longer to settle the sediment. Decant the clear supernatant liquid and use to prepare the medium.

102. DESULFOBACTER POSTGATEI MEDIUM

Solution 1:

Na₂SO₄ 3.0 g

KH₂PO₄ 0.2 g

NH₄Cl 0.3 g

NaCl 7.0 g

MgCl₂ , 6 H₂O 1.3 g

KCl 0.5 g

CaCl₂ , 2 H₂O 0.15 g

Distilled water 870.0 ml

Solution 2:

Trace element solution SL-10 (see below) 1.0 ml

Solution 3:

NaHCO₃ 5.0 g

Distilled water 100.0 ml

Solution 4:

Na-acetate , 3H₂O 2.5 g

Distilled water 10.0 ml

Solution 5:

Vitamin solution (see below) 10.0 ml

Solution 6:

Na₂S x 9 H₂O 0.4 g

Distilled water 10.0 ml

Trace element solution SL-10:

HCl (25%; 7.7M) 10.0 ml

FeCl₂ , 4H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ , 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ , 6 H₂O 190.0 mg

CuCl₂ , 2 H₂O 2.0 mg

NiCl₂ , 6 H₂O 24.0 mg

Na₂MoO₄ x 2H₂O 36.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

Ca DL-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

pH 7.1-7.4

Solution 1 is boiled before sterilization for a few minutes being flushed with gas mixture of 80% N₂ and 20% CO₂ with sodium bicarbonate added until an equilibrium pH of 6.9-7.1. Solution 1 is autoclaved under this gas mixture at 121°C for 15 min. Solution 2 is autoclaved under 100% N₂ at 121°C for 15 min. Vitamin solution is filter sterilized.

103. DESULFOBULBUS MEDIUM

Solution 1:

Na₂SO₄ 3.0 g

KH₂PO₄ 0.2 g

NH₄Cl 0.3 g

NaCl 1.0 g

MgCl₂ , 6 H₂O0.4 g

KCl 0.5 g

CaCl₂ , 2 H₂O 0.15 g

Distilled water 870.0 ml

Solution 2:

Trace element solution SL-10 (see below) 1.0 ml

Solution 3:

NaHCO₃ 5.0 g

Distilled water 100.0 ml

Solution 4:

Na-propionate 1.5 g

Distilled water 10.0 ml

Solution 5:

Vitamin solution (see below) 10.0 ml

Solution 6:

Na₂S , 9 H₂O 0.4 g

Distilled water 10.0 ml

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ , 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ , 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ , 6 H₂O 190.0 mg

CuCl₂ , 2 H₂O 2.0 mg

NiCl₂ x 6 H₂O 24.0 mg

Na₂MoO₄ , 2 H₂O 36.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

Ca DL-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

pH 7.1-7.4

Solution 1 is boiled before sterilization for a few minutes being flushed with gas mixture of 80% N₂ and 20% CO₂ with sodium bicarbonate added until an equilibrium pH of 6.9-7.1. Solution 1 is autoclaved under this gas mixture at 121°C for 15 min. Solution 2 is autoclaved under 100% N₂ at 121°C for 15 min. Vitamin solution is filter sterilized.

104. MACROMONAS MEDIUM 1

Na-acetate 1.0 g

CaCl₂ , 2 H₂O 0.1g

Casein acidic hydrolysate 0.1 g

Yeast extract 0.1 g

FeS or CaS 0.2 g

Agar 1.0 g

Distilled water 1000.0 ml

pH 7.2-7.4

Prepare suspension of FeS separately from the equimolar solutions of Na₂S x 9H₂O and FeSO₄, wash with freshly boiled distilled water under the flow of inert gas, sterilize separately from the main me-dium under inert gas at 111°C for 30 min. Main medium autoclave at 121°C for 15 min.

105. MACROMONAS MEDIUM 2

(NH₄)₂SO₄ 0.5 g

MgSO₄ , 7 H₂O 0.2 g

Casein acidic hydrolysate 1.0 g

Na-acetate , 3 H₂O 1.0 g, or succinate, or benzoate 0.5 g

Na₂S₂O₃ , 5 H₂O 0.5 g

Catalase 2.0 mg

Vitamin solution (see below) 1.0 ml

Distilled water 1000.0 ml

Vitamin solution:

Biotin 200.0 mg

Folic acid 20.0 mg

Pyridoxine-HCl 100.0 mg

Thiamine-HCl 50.0 mg

Riboflavin 100.0 mg

Nicotinic acid 50.0 mg

DL-Pantothenic acid 50.0 mg

Vitamin B₁₂ 1.0 mg

p-Aminobenzoic acid 50.0 mg

Distilled water 1000.0 ml

pH 7.2-7.4

Autoclave at 121°C for 15 min. Sterilize catalase and vitamins separately from the main medium by filtration. Thiosulfate should also better be autoclaved separately at 121°C for 15 min.

106. BEGGIATOA MEDIUM 1

(NH₄)₂SO₄ 500.0 mg

MgSO₄ , 7 H₂O 100.0 mg

CaCl₂ , 2 H₂O 50.0 mg

Na-lactate 500.0 mg

Na₂S₂O₃ , 5 H₂O 500.0 mg

K₂HPO₄ 110.0 mg

KH₂PO₄ 85.0 mg

FeCl₃ , 6 H₂O 2.0 mg

EDTA 3.0 mg

Vitamin solution (see below) 1.0 ml

Buffer HEPES 0.01 m

Distilled water 1000.0 ml

Vitamin solution:

Biotin 200.0 mg

Folic acid 20.0 mg

Pyridoxine-HCl 100.0 mg

Thiamine-HCl 50.0 mg

Riboflavin 100.0 mg

Nicotinic acid 50.0 mg

DL-Pantothenic acid 50.0 mg

Vitamin B₁₂ 1.0 mg

p-Aminobenzoic acid 50.0 mg

Distilled water 1000.0 ml

pH 7.2-7.5 (adjust with NaOH)

Sterilize at 121°C for 15 min.

Sterilize vitamins (by filtration); main medium, thiosulfate and lactate each separately (autoclave at 121°C for 15 min) and add into the main medium prior to inoculation.

107. DESULFOSARCINA MEDIUM

Solution 1:

Na₂SO₄ 3.0 g

KH₂PO₄ 0.2 g

NH₄Cl 0.3 g

NaCl 13.5 g

MgCl₂ x 6 H₂O 2.0 g

KCl 0.5 g

CaCl₂ x 2 H₂O 0.15 g

Distilled water 870.0 ml

Solution 2:

Trace element solution (see below) 1.0 ml

Solution 3:

NaHCO₃ 5.0 g

Distilled water 100.0 ml

Solution 4:

Na-benzoate 0.6 g

Na-lactate 1.0 g

Distilled water 10.0 ml

Solution 5:

Vitamin solution (see below) 10.0 ml

Solution 6:

Na₂S x 9 H₂O 0.4 g

Distilled water 10.0 ml

Trace element solution:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ x 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ x 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ x 6 H₂O 190.0 mg

CuCl₂ x 2 H₂O 2.0 mg

NiCl₂ x 6 H₂O 24.0 mg

Na₂MoO₄ x 2 H₂O 36.0 mg

Na₂SeO₄ x 5 H₂O 3.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

Ca DL-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

pH 7.1-7.4

Solution 1 is boiled before sterilization for a few minutes being flushed with gas mixture of 80% N₂ and 20% CO₂ with sodium bicarbonate added until an equilibrium pH of 6.9-7.1. Solution 1 is autoclaved under this gas mixture at 121°C for 15 min. Solution 2 is autoclaved under 100% N₂ at 121°C for 15 min. Vitamin solution is filter sterilized.

108. BEGGIATOA MEDIUM 2

MgCl₂ x 6 H₂O 50.0 mg

CaCl₂ x 2 H₂O 30.0 mg

Na-lactate 500.0 mg

Na₂S₂O₃ x 5 H₂O 500.0 mg

NaH₂PO₄ 125.0 mg

KCl 125.0 mg

NaHCO₃ 125.0 mg

Na₂SO₄ 500.0 mg

NaNO₃ 620.0 mg

Vitamin solution (see below) 1.0 ml

Trace element solution according to Hogland: (see below) 1.0 ml

Distilled water 1000.0 ml

Vitamin solution:

Biotin 200.0 mg

Folic acid 20.0 mg

Pyridoxine-HCl 100.0 mg

Thiamine-HCl 50.0 mg

Riboflavin 100.0 mg

Nicotinic acid 50.0 mg

DL-Pantothenic acid 50.0 mg

Vitamin B₁₂ 1.0 mg

p-Aminobenzoic acid 50.0 mg

Distilled water 1000.0 ml

Trace element solution according to Hogland:

EDTA 5.0 g

FeSO₄ x 7 H₂O 2.0 g

ZnSO₄ x 7 H₂O 100.0 mg

MnCl₂ x 4 H₂O 30.0 mg

H₃BO₃ 300.0 mg

CoCl₂ x 6 H₂O 200.0 mg

CuCl₂ 10.0 mg

NiCl₂ x 6 H₂O 20.0 mg

Na₂MoO₄ x 2 H₂O 20.0 mg

Distilled water 1000.0 ml

pH 7.5 (adjust with 1% HCl)

To prepare the trace element solution, preliminarily acidify water to pH 3.0-4.0 with HCl. Sterilize thiosulfate, lactate, bicarbonate, trace elements and vitamins separately and add to the main medium prior to inoculation. Lactate and thiosulfate can be more conveniently prepared as 10% solutions; bicarbonate, as 5% solution. Sterilize the vitamin solution by filtration, others solutions and base medium by autoclaving at 121°C for 15 min.

109. DESULFOVIBRIO MEDIUM

Solution 1:

K₂HPO₄ 0.5 g

NH₄Cl 1.0 g

Na₂SO₄ 1.0 g

CaCl₂ x 2 H₂O 0.1 g

MgSO₄ x 7 H₂O 2.0 g

Na DL-lactate 2.0 g

Yeast extract 1.0 g

Resazurin 1.0 g

Distilled water 980.0 ml

Solution 2:

FeSO₄ x 7 H₂O 0.5 g

Distilled water 10.0 ml

Solution 3:

Na-thioglycollate 0.1 g

Ascorbic acid 0.1 g

Distilled water 10.0 ml

Solution 4:

Trace element solution (see below) 1.0 ml

Trace element solution:

FeCl₂ x 4 H₂O 1.5 g

ZnCl₂ 68.0 mg

MnCl₂ x 4 H₂O 100.0 ml

H₃BO₃ 62.0 mg

CoCl₂ x 6 H₂O 120.0 ml

Na₂MoO₄ x 2 H₂O 24.0 mg

HCl, 0.05 M 1000.0 ml

pH 7.4-7.8

Autoclave at 121°C for 15 min under an atmosphere of N₂. Solution 1 is boiled before sterilization, being blown with N₂.

110. DESULFOVIBRIO MEDIUM WITH 2% NaCl

Solution 1:

K₂HPO₄ 0.5 g

NH₄Cl 1.0 g

NaCl 20 g

Na₂SO₄ 1.0 g

CaCl₂ x 2 H₂O 0.1 g

MgSO₄ x 7 H₂O 2.0 g

Na DL-lactate 2.0 g

Yeast extract 1.0 g

Resazurin 1.0 mg

Distilled water 980.0 ml

Solution 2:

FeSO₄ x 7 H₂O 0.5 g

Distilled water 10.0 ml

Solution 3:

Na-thioglycollate 0.1 g

Ascorbic acid 0.1 g

Distilled water 10.0 ml

Solution 4:

Trace element solution (see below) 1.0 ml

Trace element solution:

FeCl₂ x 4 H₂O 1.5 g

ZnCl₂ 68.0 mg

MnCl₂ x 4 H₂O 100.0 mg

H₃BO₃ 62.0 mg

CoCl₂ x 6 H₂O 120.0 mg

Na₂MoO₄ x 2 H₂O 24.0 mg

HCl, 0.05 M 1000.0 ml

pH 7.4-7.8

Autoclave at 121°C for 15 min under an atmosphere of N₂. Solution 1 is boiled before sterilization, being blown with N₂.

111. DESULFOVIBRIO MEDIUM WITH 3% NaCl

Solution 1:

K₂HPO₄ 0.5 g

NH₄Cl 1.0 g

NaCl 30 g

Na₂SO₄ 1.0 g

CaCl₂ x 2 H₂O 0.1 g

MgSO₄ x 7 H₂O 2.0 g

Na DL-Lactate 2.0 g

Yeast extract 1.0 g

Resazurin 1.0 mg

Distilled water 980.0 ml

Solution 2:

FeSO₄ x 7 H₂O 0.5 g

Distilled water 10.0 ml

Solution 3:

Na-thioglycollate 0.1 g

Ascorbic acid 0.1 g

Distilled water 10.0 ml

Solution 4:

Trace element solution (see below) 1.0 ml

Trace element solution:

FeCl₂ x 4 H₂O 1.5 g

ZnCl₂ 68.0 mg

MnCl₂ x 4 H₂O 100.0 mg

H₃BO₃ 62.0 mg

CoCl₂ x 6 H₂O 120.0 mg

Na₂MoO₄ x 2 H₂O 24.0 mg

HCl, 0.05 M 1000.0 ml

pH 7.4-7.8

Autoclave at 121°C for 15 min under an atmosphere of N₂. Solution 1 is boiled before sterilization, being blown with N₂.

112. GLUCOSE YEAST EXTRACT AGAR

Glucose 20.0 g

Yeast extract 10.0 g

CaCO₃ 20.0 g

Agar 17.0 g

Distilled water 1000.0 ml

Sterilize at 111°C for 30 min.

113. DESULFOVIBRIO BAARSII MEDIUM

Solution 1:

Na₂SO₄ 3.0 g

KH₂PO₄ 0.2 g

NH₄Cl 0.3 g

NaCl 1.0 g

MgCl₂ x 6 H₂O 0.4 g

KCl 0.5 g

CaCl₂ x 2 H₂O 0.15 g

Distilled water 870.0 ml

Solution 2:

Trace element solution SL-10 (see below) 1.0 ml

Solution 3:

NaHCO₃ 5.0 g

Distilled water 100.0 ml

Solution 4:

Na-butyrate 0.7 g

Na-caproate 0.3 g

Na-octanoate 0.15 g

Distilled water 10.0 ml

Solution 5:

Vitamin solution (see below) 10.0 ml

Solution 6:

Na₂S x 9 H₂O 0.4 g

Distilled water 10.0 ml

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ x 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ x 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ x 6 H₂O 190.0 mg

CuCl₂ x 2 H₂O 2.0 mg

NiCl₂ x 6 H₂O 24.0 mg

Na₂MoO₄ x 2 H₂O 36.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

Ca DL-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

pH 6.8-7.0

Solution 1 is boiled before sterilization for a few minutes being flushed with gas mixture of 80% N₂ and 20% CO₂ with sodium bicarbonate added until an equilibrium pH of 6.9-7.1. Solution 1 is autoclaved under this gas mixture at 121°C for 15 min. Solution 2 is autoclaved under 100% N₂ at 121°C for 15 min. Vitamin solution is filter sterilized.

114. AZOSPIRILLUM BRASILIENSE MEDIUM 2

Ca-malate 10.0 g or

Glucose 20.0 g

K₂HPO₄ 0.1 g

KH₂PO₄ 0.2 g

MgSO₄ x 7 H₂O 0.5 g

FeCl₃ x 6 H₂O 0.1 g

CaCl₂ x 2 H₂O 0.02 g

Na₂MoO₄ x 2 H₂O 0.02 mg

Yeast extract 0.1 g

Agar 15.0 g

Distilled water 1000.0 ml

pH 6.9

Autoclave at 111°C for 30 min.

115. METHANOBACTERIUM MEDIUM

KH₂PO₄ 0.5 g

MgSO₄ x 7 H₂O 0.4 g

NaCl 0.4 g

NH₄Cl 0.4 g

CaCl₂ x 2 H₂O 0.05 g

Trace element solution SL-10 (see below) 1.0 ml

Vitamin solution (see below) 5.0 ml

Yeast extract 1.0 g

Na-acetate 1.0 g

Na-formate 2.0 g

NaHCO₃ 4.0 g

Resazurin 1.0 mg

L-Cysteine-HCl 0.5 g

Na₂S x 9 H₂O 0.5 g

Fatty acid mixture (see below) 20.0 ml

Sludge water: (see below) 50.0 ml

Distilled water 940.0 ml

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ x 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ x 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ x 6 H₂O 190.0 mg

CuCl₂ x 2 H₂O 2.0 mg

NiCl₂ x 6 H₂O 24.0 mg

Na₂MoO₄ x 2 H₂O 36.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine (B₂) 10.0 mg

Riboflavin (B₁) 5.0 mg

Pantotenoic acid 5.0 mg

p-Aminobenzoic acid 5.0 mg

Thiamine-HCl 5.0 mg

Nicotinic acid 5.0 mg

Cyanocobalamin (B₁₂) 0.1 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

Fatty acid mixture:

Valeric acid 0.5 g

Isovaleric acid 0.5 g

α-Methylbutyric acid 0.5 g

Isobutyric acid 0.5 g

Distilled water 20.0 ml

pH 7.5 (adjust with concentrated NaOH)

Sludge water: to sludge from an anaerobic digester add 0.4% yeast extract and after gassing with

N₂ gas for a few minutes incubate it at 37°C for 24 hours. Then centrifuge the sludge at 13,000 g and autoclave the resulting clear supernatant under N₂ gas at 121°C for 15 min (pH 6.7-7.0). Base medium and trace element solution autoclave at 121°C for 15 min. Vitamin solution is filter sterilized. Prepare the medium anaerobically under a gas atmosphere of 80% H₂ and 20% CO₂.

116. THERMUS RUBER MEDIUM

Pepton 5.0 g

Yeast extract 1.0 g

Starch (soluble) 1.0 g

Agar 12.0 g

Distilled water 1000.0 ml

pH 8.0

Sterilize at 121°C for 15 min.

117. METHANOSARCINA MEDIUM

Solution 1:

NaCl 0.9 g

MgCl₂ x 6 H₂O 0.2 g

CaCl₂ x 2 H₂O 0.1 g

NH₄Cl 1.0 g

Yeast extract 2.0 g

Resazurin 10.0 mg

Methanol 10% 10.0 ml

Trace element solution (see below) 10.0 ml

Vitamin solution (see below) 5.0 ml

Distilled water 965.0 ml

Solution 2 (reducing agents):

L-Cysteine-HCl 0.5 g

Na₂S x 9 H₂O 0. 5 g

Distilled water 10.0 ml

Buffer solutions:

a) K₂HPO₄ 29.0 g

Distilled water 100.0 ml

b) KH₂PO₄ 15.0 g

Distilled water 100.0 ml

Trace element solution:

Nitrilotriacetic acid 12.8 g

FeSO₄ x 7 H₂O 0.1 g

MnCl₂ x 6 H₂O 0.1 g

CoCl₂ x 2 H₂O 0.17 g

CaCl₂ x 2 H₂O 0.1 g

ZnCl₂ 0.1 g

CuCl₂ x 2 H₂O 0.02 g

H₃BO₃ 0.01 g

Na₂MoO₄ x 2 H₂O 0.01 g

NaCl 1.0 g

Na₂SeO₄ 0.017 g

Distilled water 1000.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine 10.0 mg

Riboflavin 5.0 mg

Pantotenoic acid 5.0 mg

p-Aminobenzoic acid 5.0 mg

Thiamine-HCl 5.0 mg

Nicotinic acid 5.0 mg

Vitamin B₁₂ 0.1 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

pH 7.2-7.4

Prepare medium in anaerobic conditions, blowing through with N₂without O₂ up to sterilization. Solutions of reducing agents (10.0 ml) and of buffer (per 1 ml) add to base medium after separate sterilization. Base medium, trace elements, buffer solutions and reducing agents autoclave at 121°C for 15 min. Vitamin solution is filter sterilized.

118. PLATE COUNT AGAR

Tryptone 5.0 g

Yeast extract 2.5 g

Glucose 1.0 g

Agar 15.0 g

Distilled water 1000.0 ml

pH 7.0±0.2

Sterilize at 121°C for 15 min.

119. NITROSOLOBUS MEDIUM 2

(NH₄)₂SO₄ 0.5 g

KH₂PO₄ 0.2 g

CaCl₂ x 2 H₂O 0.02 g

MgSO₄ x 7 H₂O 0.04 g

EDTA FeNa 3.8% 0.1 ml

Phenol red 0.05% 2.0 ml

Distilled water 1000.0 ml

pH 8.0 (adjust with 6% Na₂CO₃)

Autoclave at 121°C for 15 min.

120. MANNITOL AGAR WITH PEPTONE

Yeast extract 5.0 g

Mannitol 25.0 g

Peptone 3.0 g

Agar 15.0 g

Distilled water 1000.0 ml

pH 5.0-6.0

Sterilize at 121°C for 15 min.

121. THIOBACILLUS THIOOXIDANS MEDIUM (WAKSMAN MEDIUM)

(NH₄)₂SO₄ 300.0 mg

MgSO₄ x 7 H₂O 500.0 mg

KH₂PO₄ 3.5 g

CaCl₂ x 2 H₂O 250.0 mg

FeSO₄ x 7 H₂O 18.0 mg

Finely dispersed sulfur 5.0 g

Distilled water 1000.0 ml

pH 4.5

Autoclave at 121°C for 15 min.

122. THIOBACILLUS FERROOXIDANS MEDIUM (LEATHEN MEDIUM)

(NH₄)₂SO₄ 150.0 mg

KCl 50.0 mg

MgSO₄ , 7 H₂O 500.0 mg

KH₂PO₄100.0 mg

Ca(NO₃)₂ , 4 H₂O 10.0 mg

Distilled water 1000.0 ml

Autoclave at 121°C for 15 min.

After sterilization of the medium, add 10.0 ml of 10% FeSO₄ , 7 H₂O preliminarily acidified to pH 3.5 and autoclaved separately in sealed ampoules under nitrogen. pH 4.0 (adjust after sterilization and addition of iron solution).

123. MEDIUM FOR MARINE NITROBACTER

Solution 1 (see below) 0.5 ml

Solution 2 (see below) 0.5 ml

Solution 3 (see below) 1.0 ml

Solution 4 (see below) 0.5 ml

Solution 5 (see below) 0.5 ml

Solution 6 (see below) 0.1 ml

Distilled water 300.0 ml

Sea water 700.0 ml

Solution 1:

CaCl₂ 2.0 g

Distilled water 100.0 ml

Solution 2:

MgSO₄ x 7 H₂O 20.0 g

Distilled water 100.0 ml

Solution 3:

Chelated iron 0.1 g

Distilled water 100.0 ml

Solution 4:

Na₂MoO₄ 0.1 g

MnCl₂ x 4 H₂O 0.2 g

CoCl₂ x 6 H₂O 0.0 2 g

ZnSO₄ x 7 H₂O 0.1 g

CuSO₄ x 5 H₂O 0.02 g

Distilled water 1000.0 ml

Solution 5:

NaNO₃ 41.4 g

Distilled water 100.0 ml

Solution 6:

K₂HPO₄ 1.74 g

Distilled water 100.0 ml

pH 8.6 (adjust with NaOH or KOH)

Autoclave all solutions separately at 121°C for 15 min. Leave the medium to stand for 2 – 3 days so that pH can adjust itself to pH 7.4 - 7.6.

124. AZOSPIRILLUM MEDIUM

Solution 1:

Yeast extract 0.05 g

FeSO₄ x 7 H₂O 0.01 g

K₂HPO₄ 0.25 g

Na₂MoO₄ x 2 H₂O 1.0 mg

MnSO₄ x H₂O 2.0 mg

MgSO₄ x 7 H₂O 0.2 g

NaCl 0.1 g

CaCl₂ x 2 H₂O 0.02 g

(NH₄)₂SO₄ 1.0 g

Biotin 0.1 mg

Bromothymol blue 25.0 mg

Distilled water 950.0 ml

Solution 2:

Glucose 20% 25.0 ml

Solution 3:

Na-malate 20% 25.0 ml

Dissolve bromothymol blue in diluted KOH before adding into the medium.

pH 7.1

Autoclave solutions separately at 121°C for 15 min.

125. NITROSOLOBUS MEDIUM 3

(NH₄)₂SO₄ 2.0 g

CaCl₂ x 2 H₂O 20.0 mg

MgSO₄ x 7 H₂O 200.0 mg

Chelated iron 1.0 mg

Na₂MoO₄ x 2 H₂O 100.0 mg

MnCl₂ x 4 H₂O 200.0 mg

CoCl₂ x 6 H₂O 2.0 mg

ZnSO₄ x 7 H₂O 100.0 mg

K₂HPO₄ 15.9 mg

CuSO₄ x 5 H₂O 20.0 mg

Distilled water 1000.0 ml

pH 7.5-7.8

Autoclave at 121°C for 15 min.

126. DESULFOCOCCUS NIACINI MEDIUM

Solution 1:

Na₂SO₄ 3.0 g

KH₂PO₄ 0.2 g

NH₄Cl 0.3 g

NaCl 13.5 g

MgCl₂ x 6 H₂O 2.2 g

KCl 0.5 g

CaCl₂ x 2 H₂O 0.15 g

Distilled water 870.0 ml

Solution 2:

Trace element solution SL-10 (see below) 1.0 ml

Solution 3:

NaHCO₃ 5.0 g

Distilled water 100.0 ml

Solution 4:

Na-nicotinate 5.0 mM

Solution 5:

Vitamin solution (see below) 10.0 ml

Solution 6:

Na₂S x 9 H₂O 0.4 g

Distilled water 10.0 ml

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ x 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ x 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ x 6 H₂O 190.0 mg

CuCl₂ x 2 H₂O 2.0 mg

NiCl₂ x 6 H₂O 24.0 mg

Na₂MoO₄ x 2 H₂O 36.0 mg

Na₂SeO₃ x 5 H₂O 40.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

Ca DL-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

Solution 1 is boiled before sterilization for a few minutes being flushed with gas mixture of 80% N₂ and 20% CO₂ with sodium bicarbonate added until an equilibrium pH of 6.9-7.1. Solution 1 is autoclaved under this gas mixture at 121°C for 15 min. Solution 2 is autoclaved under 100% N₂ at 121°C for 15 min. Trace element solution is autoclaved at 121°C for 15 min. Vitamin solution is filter sterilized.

Final pH of the complete medium 7.4

127. SELENITE CONTROL MEDIUM

Solution 1:

Peptone 5.0 g

Na₂HPO₄ 7.0 g

NaH₂PO₄ 3.0 g

Lactose 4.0 g

Distilled water 960.0 ml

pH 6.9-7.1

Solution 2:

10% Na₂SeO₃ x 5 H₂O 40.0 ml

Autoclave at 111°C for 30 min.

128. HIRSCH MEDIUM

KH₂PO₄ 1.36 g

Na₂HPO₄ 2.15 g

MgSO₄ x 7 H₂O 0.2 g

MnSO₄ x 5 H₂O 1.05 g

(NH₄)₂SO₄ 0.5 g

CaCl₂ x 2 H₂O 5.97 mg

Na₂MoO₄ x 2 H₂O 1.5 mg

Methanol 5.0 ml

Distilled water to 1000.0 ml

pH 7.0

Autoclave at 121°C for 15 min. Sterilize methanol by filtration and add to the medium after autoclaving.

129. CONTROL MEDIUM C-1

KH₂PO₄ 2.0 g

K₂HPO₄ 2.0 g

MgSO₄ x 7 H₂O 0.2 g

NaCl 5.0 g

Raffinose 2.0 g

Bromothymol blue (1.6% alkaline) 2.0 ml

Crystal violet (0.01%) 20.0 ml

Urea (50%) 4.0 ml

Distilled water 975.0 ml

Autoclave at 111°C for 30 min. Urea solution is filter sterilized.

130. GLUCOSE POTATO AGAR

Potato 200.0 g

Glucose 10.0 g

Agar 20.0 g

Tap water 1000.0 ml

pH 7.0

Boil 200 g scrubbed and sliced potatoes in 1000.0 ml water for 1 hour, filter cold through a cotton-gauze filter, add water to the initial volume, add glucose and agar. Do not use new potatoes.

Sterilize at 111°C for 30 min.

131. MEDIUM Q MOD FOR FRANKIA

K₂HPO₄ 300.0 mg

NaH₂PO₄ 200.0 mg

MgSO₄ x 7 H₂O 200.0 mg

KCl 200.0 mg

Yeast extract 500.0 mg

Peptone 5.0 g

Glucose 10.0 g

Fe-citrate 1.0 ml

Trace element solution (see below) 1.0 ml

CaCO₃ 100.0 mg

Tween-80 2.0 g

Agar 15.0 g

Distilled water 1000.0 ml

Trace element solution:

H₃BO₃ 1.5 g

MnSO₄ x 7 H₂O 0.8 g

ZnSO₄ x 7 H₂O 0.6 g

CuSO₄ x 5 H₂O 0.1 g

(NH₄)₆Mo₇O₂₄ x 4 H₂O 0.2 g

CoSO₄ 0.001 g

Distilled water 1000.0 ml

pH 7.0

Autoclave at 111°C for 30 min.

132. THIOBACILLUS FERROOXIDANS MEDIUM 9K

Solution 1:

KCl 0.1 g

MgSO₄ x 7 H₂O 0.5 g

K₂HPO₄ 0.5 g

Ca(NO₃)₂ x 4 H₂O 0.01 g

Distilled water 700.0 ml

Solution 2:

(NH₄)₂Fe(SO₄)₂ 63.0 g

H₂SO₄ (10 N) 1.0 ml

Distilled water 300.0 ml

pH 3.5

Autoclave solution 1 at 121°C for 15 min, solution 2 at 111°C 30 min. Mix the solutions before inoculation.

133. COLBY AND ZATMAN MEDIUM WITH METHANOL

K₂HPO₄ 1.2 g

KH₂PO₄ 0.62 g

CaCl₂ x 6 H₂O 0.05 g

MgSO₄ x 7 H₂O 0.2 g

NaCl 0.1 g

FeCl₃ x 6 H₂O 1.0 mg

(NH₄)₂SO₄ 0.5 g

CuSO₄ x 5 H₂O 5.0 µg

MnSO₄ x 5 H₂O 10.0 µg

Na₂MoO₄ x 2 H₂O 10.0µg

H₃BO₃ 10.0 µg

ZnSO₄ x 7 H₂O 70.0 µg

CoCl₂ x 6 H₂O 5.0µg

Purified agar 15.0 g

Methanol 1.0 ml

Distilled water 1000.0 ml

pH 6.8

Autoclave at 121°C for 15 min. Methanol is filter sterilized.

134. METHYLOTROPH MEDIUM 1

KH₂PO₄ 2.0 g

(NH₄)₂SO₄ 2.0 g

MgSO₄ x 7 H₂O 0.025 g

NaCl 0.5 g

FeSO₄ x 7 H₂O 0.02 g

Methanol 5.0 ml

Distilled water 1000.0 ml

pH 7.0

Autoclave at 121°C for 15 min. Methanol is filter sterilized. Methanol may be replacement 30 ml 10 % solution of methylamine, autoclave at 111°C for 30 min.

135. METHYLOTROPH MEDIUM 2

KH₂PO₄ 0.8 g

Na₂HPO₄ x 12 H₂O 3.0 g

(NH₄)₂SO₄ 0.8 g

MgSO₄ x 7 H₂O 0.1 g

FeSO₄ x 7 H₂O 10.0 mg

Methanol 5.0 ml

Trace element solution (see below) 1.0 ml

Distilled water 1000.0 ml

Trace element solution:

CaCl₂ x 2 H₂O 1.25 g

CoCl₂ x 6 H₂O 1.25 g

MnSO₄ x 4 H₂O 0.1 g

ZnSO₄ x 7 H₂O 1.25 g

Na₂MoO₄ x 2 H₂O 50.0 mg

CuCl₂ x 2 H₂O 250.0 mg

Distilled water 250.0 ml

pH 7.0-7.2

Autoclave at 121°C for 15 min. Methanol is filter sterilized. Methanol may be replacement 30 ml 10 % solution of methylamine, autoclave at 111°C for 30 min.

136. MEDIUM FOR PERCHLORATE-REDUCING BACTERIA

Solution 1:

NH₄Cl 0.1 g

NaCl 0.02 g

KH₂PO₄ 0.1 g

K₂HPO₄ 0.4 g

MgSO₄ x 7 H₂O 0.1 g

Tap water 1000.0 ml

Solution 2:

HCl, to dissolve the precipitate in solution 1

Solution 3:

pH 6.9-7.2 (adjust with 5% NaOH)

Solution 4:

5% NH₄ClO₄ 5.0 ml

Solution 5:

5% Na-acetate 5.0 ml

Solution 6:

Trace element solution according to Hogland (see below) 0.5 ml

Solution 7:

Vitamin B₁₂ (dispensary solution) 0.2 ml

Solution 8:

96° ethanol 0.5 ml

Trace element solution according to Hogland:

EDTA 5.0 g

FeSO₄ x 7 H₂O 2.0 g

ZnSO₄ x 7 H₂O 100.0 mg

MnCl₂ x 4 H₂O 30.0 mg

H₃BO₃ 300.0 mg

CoCl₂ x 6 H₂O 200.0 mg

CuCl₂ x 2 H₂O 10.0 mg

NiCl₂ x 6 H₂O 20.0 mg

Na₂MoO₄ x 2 H₂O 20.0 mg

Distilled water 1000.0 ml

Sterilize ethanol by filtration, others solutions separately autoclave at 121°C for 15 min. Add solutions and additions to the main medium in the order of their enumeration.

137. MEDIUM FOR CHROMATE-REDUCING BACTERIA

Solution 1:

NH₄Cl 0.3 g

NaCl 0.1 g

KH₂PO₄ 0.5 g

K₂HPO₄ 0.3 g

MgSO₄ x 7 H₂O 0.1 g

CaCO₃ 0.05 g

FeCl₃ x 6 H₂O 0.05 g

K₂CrO₄ 0.1 g

Paper-filtered pond water 900.0 ml

Solution 2:

Peptone meat broth 100.0 ml

Solution 3:

5% Na-acetate 5.0 ml

Solution 4:

Trace element solution according to Hogland (see below) 0.5 ml

Solution 5:

Vitamin B₁₂ (dispensary solution) 0.2 ml

Trace element solution according to Hogland:

EDTA 5.0 g

FeSO₄ x 7 H₂O 2.0 g

ZnSO₄ x 7 H₂O 100.0 mg

MnCl₂ x 4 H₂O 30.0 mg

H₃BO₃ 300.0 mg

CoCl₂ x 6 H₂O 200.0 mg

CuCl₂ x 2 H₂O 10.0 mg

NiCl₂ x 6 H₂O 20.0 mg

Na₂MoO₄ x 2 H₂O 20.0 mg

Distilled water 1000.0 ml

pH 7.0-7.2

Autoclave solutions separately at 121°C for 15 min. Add solutions and additions to the main medium in the order of their enumeration.

138. LARSEN PHOTOTROPH MEDIUM

NH₄Cl 0.5 g

KH₂PO₄ 1.0 g

MgCl₂ 0.5 g

CaCl₂ x 2 H₂O 0.1 g

Trace element solution SL-12B (see below) 1.0 ml

NaHCO₃ 5.0 g

Na-acetate x 3 H₂O 2.0 g

Na₂S₂O₃ 0.4-0.6 g

Na₂S x 9 H₂O 0.1 g

Fe-citrate Traces

Distilled water 1000.0 ml

Trace element solution SL-12B:

EDTA Na 3.0 g

FeSO₄ x 7 H₂O 1.1 g

CoCl₂ x 6 H₂O 190.0 mg

MnCl₂ x 2 H₂O 50.0 mg

ZnCl₂ 42.0 mg

NiCl₂ x 6 H₂O 24.0 mg

Na₂MoO₄ x 2 H₂O 18.0 mg

H₃BO₃ 300.0 mg

CuCl₂ x 2 H₂O 2.0 mg

Distilled water 1000.0 ml

pH of the trace element solution 6.0

pH 8.4

Autoclave solutions separately at 121°C for 15 min.

139. POSTGATE MEDIUM B FOR SULFATE REDUCERS

Solution 1:

NaCl 1.0 g

KH₂PO₄ 0.5 g

NH₄Cl 1.0 g

CaSO₄ x 2 H₂O 1.0 g

MgSO₄ x 7 H₂O 2.0 g

Na-lactate 3.5 g

Yeast extract 1.0 g

Tap water 980.0 ml

Dissolve ingredients and sparge medium with O₂-free N₂ gas for 10-15 minutes

Solution 2:

Ascorbic acid 1.0 g

Thioglycolic acid 1.0 g

Distilled water 10.0 ml

Solution 3:

FeSO₄ x 7 H₂O 0.5 g

Distilled water 10.0 ml

pH 7.4

Autoclave solution 1 at 121°C for 15 min, solutions 2 and 3 separately autoclave at 111°C for 30 min.

Boil 200 g scrubbed and sliced potatoes in 1000.0 ml water for 1 hour, filter cold through a cotton-gauze filter, add water to the initial volume, add glucose and agar. Do not use new potatoes.

147. HYPHOMICROBIUM MEDIUM

NaNO₃ 1.0 g

NaCl 0.5 g

K₂HPO₄ 1.0 g

MgSO₄ x 7 H₂O 0.2 g

Methanol 5.0-10.0 ml

Distilled water 1000.0 ml

pH 6.8-7.0

Autoclave at 121°C for 15 min. Methanol is filter sterilized.

Grow in an exicator in the presence of methanol vapors.

149. CLAVIBACTER XYLI MEDIUM

Solution 1:

Flour agar 5.0 g

Papaya hydrolysate of soybean meal 8.0 g

K₂HPO₄ 1.0 g

KH₂PO₄ 1.0 g

MgSO₄ x 7 H₂O 0.2 g

Distilled water 965.0 ml

Solution 2:

Bovine hemine chloride (0.1% in 0.05 N NaOH) 15.0 ml

Solution 3:

Bovine serum albumin, fraction 5 (20%) 10.0 ml

Solution 4:

Glucose (50%) 1.0 ml

Solution 5:

Cysteine (10%) 10.0 ml

pH 6.6

Autoclave solution 1 at 121°C for 15 min, cool to 50 C and add the filter-sterilized solutions 2-5.

150. GETCHINSON MEDIUM WITH FILTER PAPER

K₂HPO₄ 1.3 g

MgSO₄ x 7 H₂O 0.3 g

CaCl₂ x 6 H₂O 0.1 g

FeCl₃ x 6 H₂O 0.01 g

NaNO₃ 2.5 g

Distilled water 1000.0 ml

pH 7.2-7.3.

Autoclave at 121°C for 15 min. Cut filter paper into strips, sterilize by dry heat and immerse into the medium so that they are not completely in the liquid medium.

151. MUNZ MEDIUM FOR METHANE-OXIDIZING BACTERIA

K₂HPO₄ 0.5 g

KH₂PO₄ 0.5 g

MgSO₄ x 7 H₂O 0.5 g

NH₄Cl 1.0 g

Tap water 1000.0 ml

pH 6.8

Autoclave at 121°C for 15 min.

Cultivate in the mixed atmosphere of air and methane (2:1).

152. TRYPTONE THIOGLYCOLATE MEDIUM

Solution 1:

K₂HPO₄ 5.45 g

KH₂PO₄ 1.2 g

MgSO₄ x 7 H₂O 25.0 mg

CaCl₂ x 2 H₂O 15.0 mg

FeSO₄ x 7 H₂O 10.0 mg

MnCl₂ x 4 H₂O 2.0 mg

CoCl₂ x 6 H₂O 2.5 mg

Na₂MoO₄ x 2 H₂O 2.5 mg

Peptone 2.0 g

Tryptone 2.0 g

Yeast extract 6.0 g

Na-thioglycolate 0.5 g

Distilled water 950.0 ml

Solution 2:

Glucose 20.0 g

Distilled water 50.0 ml

pH 7.5

Autoclave solution 1 at 121°C for 15 min, solution 2 at 111°C for 30 min.

153. MEDIUM P-2 FOR THERMOPHILIC ANAEROBIC BACTERIA

K₂HPO₄ 3.0 g

KH₂PO₄ 2.0 g

NH₄Cl 2.0 g

MgCl₂ x 6 H₂O 0.2 g

CaCl₂ x 2 H₂O 0.05 g

Tryptone 10.0 g

Glucose 5.0 g

Yeast extract 5.0 g

Resazurin 0.01 g

Distilled water 1000.0 ml

pH 7.0-7.2

Prepare medium anaerobically under N₂and autoclave (удалить under N₂at 121°C for 15 min. Autoclave glucose separately under N₂ at 111°C for 30 min.

154. DESULFOVIBRIO SULFODISMUTANS MEDIUM

Solution 1:

KH₂PO₄ 0.2 g

NH₄Cl 0.3 g

NaCl 1.0 g

MgCl₂ x 6 H₂O 0.4 g

KCl 0.5 g

CaCl₂ x 2 H₂O 0.15 g

Distilled water 920.0 ml

Solution 2:

Trace element solution SL-10 (see below) 1.0 ml

Solution 3:

NaHCO₃ 2.5 g

Distilled water 50.0 ml

Solution 4:

Na-acetate x 3 H₂O 0.3 g

Distilled water 10.0 ml

Solution 5:

D(+)-Biotin 10.0 mg

Ca-D(+)-Pantothenate 50.0 mg

Distilled water 1.0 ml

Solution 6:

Na₂S x 9 H₂O 0.4 g

Distilled water 10.0 ml

Solution 7:

Na₂S₂O₃ (0.5 M) 10.0 ml

pH 7.5-8.0 (adjust with NaOH)

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ x 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ x 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ x 6 H₂O 190.0 mg

CuCl₂ x 2 H₂O 2.0 mg

NiCl₂ x 6 H₂O 24.0 mg

Na₂MoO₄ x 2 H₂O 36.0 mg

Distilled water 990.0 ml

pH 7.1-7.4

Solution 1 is prepared and autoclaved anaerobically at under 80% N₂ + 20% CO₂at 121°C for 15 min. Solutions 2, 4, 6 and 7 are gassed with N₂ and autoclaved separately at 121°C for 15 min. Solution 3 (gassed with N₂ + CO₂) and solution 5 (gassed with N₂) are filter-sterilized. Solutions with 2 to 7 are added to the sterile, cooled solution 1 in the sequence as indicated. When growth has started feed culture again with same amount of solution 7. After a further two days repeat feeding once more.

155. SCHATZ AND BOVELL MEDIUM FOR HYDROGEN-OXIDIZING BACTERIA

KH₂PO₄ 1.0 g

NH₄NO₂ 1.0 g

MgSO₄ x 7 H₂O 0.2 g

FeSO₄ x 7 H₂O 0.01 g

CaCl₂ x 2 H₂O 0.01 g

NaHCO₃ 0.5 g

Agar 15.0 g

Distilled water 1000.0 ml

pH 6.8-7.2

Autoclave at 121°C for 15 min. Cultivate in a gas mixture of carbon dioxide, air and hydrogen (1:3:6).

156. MEDIUM FOR MYXOBACTERIA

Casein hydrolysate 2.5 g

Asparagine 2.5 g

K₂HPO₄ 2.0 g

NaCl 1.0 g

MgSO₄ x 7 H₂O 0.1 g

FeSO₄ x 7 H₂O 0.03 g

CaCl₂ x 2 H₂O 0.01 g

Distilled water 1000.0 ml

pH 7.4

Autoclave at 121°C for 15 min.

157. EMERSON STARCH YEAST EXTRACT AGAR

Yeast extract 4.0 g

Starch (soluble) 15.0 g

KH₂PO₄ 1.0 g

MgSO₄ x 7 H₂O 0.5 g

Agar 20.0 g

Distilled water 1000.0 ml

pH 7.0

Autoclave at 121°C for 15 min.

Prepare sterile agar slants by autoclaving at 121°C for 15 min. Cool the slants in a sloping position. Cover solid slants with 2 ml sterile tap water. Inoculate into the covering tap water.

162. ANCYLOBACTER-SPIROSOMA MEDIUM

Glucose 1.0 g

Peptone 1.0 g

Yeast extract 1.0 g

Agar 15.0 g

Distilled water 1000.0 ml

pH 7.0-7.2

Autoclave at 111°C for 30 min.

164. PD BROTH FOR FLEXIBACTER

Peptone 1.0 g

KNO₃ 100.0 mg

Yeast extract 100.0 mg

K₂HPO₄ 66.7 mg

MgSO₄ x 7 H₂O 33.3 mg

Trace element solution (see below) 1.0 ml

Distilled water 1000.0 ml

Trace element solution:

Zn SO₄ x 7 H₂O 22.0 mg

MnSO₄ x 7 H₂O 1.81 g

CuSO₄ x 5 H₂O 79.0 mg

NaBO₃ x 4 H₂O 1.0 g

(NH₄)₆Mo₇O₂₄ x 4 H₂O 9.3 g

FeSO₄ x 7 H₂O 20.0 mg

Co(NO₃)₂ x H₂O 20.0 mg

Trilon B 10.0 g

Distilled water 1000.0 ml

pH 7.0

Autoclave at 111°C for 30 min.

Dissolve components of the medium in boiling water in the specified sequence (except the indicator). Then filter through a cotton wool-gauze filter to remove the possible precipitate, bring tot he initial volume, cool, adjust pH 6.7, add the indicator.