The Elements of Bacteriological Technique: A Laboratory Guide for Medical, Dental, and Technical Students. Second Edition Rewritten and Enlarged.Eyre, J. W. H. (John William Henry)
Science
The Elements of Bacteriological Technique: A Laboratory Guide for Medical, Dental, and Technical Students. Second Edition Rewritten and Enlarged.
Eyre, J. W. H. (John William Henry)
Bacteriology -- Technique
2. Wash the agar emulsion into a tared 2-litre flask with 400 c.c.
distilled water.
3. Weigh out
Peptone 0.5 gramme.
Emulsify in 50 c.c. distilled water and add to the contents of the
flask.
4. Bubble live steam through the mixture for twenty minutes to dissolve
the agar.
5. Weigh out and mix
Dextrose 10.0 grammes.
Potassium phosphate 0.5 gramme.
Magnesium sulphate 0.2 gramme.
Potassium nitrate 0.06 gramme.
and add to the contents of the flask.
6. Adjust the weight of the medium mass to the calculated figure for one
litre (1025 grammes) by the addition of distilled water at 100°C.
7. Titrate the medium mass and adjust the reaction to +5.
8. Cool to 60° C. Clarify with egg and filter.
9. Tube in quantities of 10 c.c. and sterilise as for nutrient agar.
~Beyrinck's Solution. I.~--(_For the cultivation of nitrogen fixing
organisms._)
1. Weigh out and mix 1 gramme potassium hydrogen phosphate, 0.2 gramme
magnesium sulphate, and 0.02 gramme sodium chloride.
2. Dissolve in water 1000 c.c., in a 2-litre flask.
3. Add 1 c.c. of a one per thousand aqueous solution of ferrous
sulphate.
4. Add 1 c.c. of a one per thousand solution manganese sulphate.
5. Weigh out 20 grammes dextrose and add to the contents of the flask
(dextrose up to 40 grammes may be used for the different organisms).
6. Steam for twenty minutes, filter.
7. Tube, and sterilise as for nutrient bouillon.
~Beyrinck's Solution. II.~--(_For growth of Azobacter._)
Proceed as in preparing solution No. I, substituting mannite for
dextrose in step 5.
~Winogradsky's Solution (for Nitric Organisms).~--
1. Weigh out and mix.
Potassium phosphate 1.0 gramme
Magnesium sulphate 0.5 gramme
Calcium chloride 0.01 gramme
Sodium chloride 2.0 grammes
and dissolve in
Distilled water 1000 c.c.
2. Fill into flasks, in quantities of 20 c.c. and add to each a small
quantity of freshly washed magnesium carbonate.
3. Sterilise in the steamer at 100° C. for twenty minutes on each of
three consecutive days.
4. Add to each flask containing 20 c.c. solution, 2 c.c. of a sterile 2
per cent. solution of ammonium sulphate.
5. Incubate at 37° C. for forty-eight hours and eliminate any
contaminated culture flasks. Store the remainder for future use.
~Winogradsky's Solution (for Nitrous Organisms).~--
1. Weigh out and mix
Ammonium sulphate 1 gramme
Potassium sulphate 1 gramme
and dissolve in
Distilled water 1000 c.c.
2. Add 5 to 10 grammes basic magnesium carbonate, previously sterilised
by boiling.
3. Fill into flasks and sterilise, etc., as for previous solution.
~Silicate Jelly (Winogradsky).~--
1. Weigh out and mix
Ammonium sulphate 0.40 gramme
Magnesium sulphate 0.05 gramme
Calcium chloride 0.01 gramme
and dissolve in
Distilled water 50 c.c.
Label--Solution A.
2. Weigh out and mix
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