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
(c) In the presence of gases other than oxygen.
Temperature:
(a) Range.
(b) Optimum.
(c) Thermal death-point:
Moist: Vegetative forms.
Spores.
Dry: Vegetative forms.
Spores.
Reaction of medium.
Resistance to lethal agents:
(a) Desiccation.
(b) Light: Diffuse.
Direct.
Primary colours.
(c) Heat.
(d) Chemical antiseptics and disinfectants.
Vitality in artificial cultures.
~I. Atmosphere.~--The question as to whether the organism under
observation is (a) an obligate aerobe, (b) a facultative anaerobe, or
(c) an obligate anaerobe is roughly decided by the appearance of
cultivations in the fermentation tubes. Obvious growth in the closed
branch as well as in the bulb or in the inverted gas tube as well as in
the bulk of the medium will indicate that it is a facultative anaerobe;
whilst growth only occurring in the bulb or in the closed branch shows
that it is an obligate aerobe or anaerobe respectively. This method,
however, is not sufficiently accurate for the present purpose, and the
examination of an organism with respect to its behaviour in the absence
of oxygen is carried out as follows:
_Apparatus Required:_
Buchner's tubes.
Bulloch's apparatus.
Exhaust pump.
Pyrogallic acid.
Dekanormal caustic soda.
_Media Required:_
Glucose formate agar.
Glucose formate gelatine.
Glucose formate bouillon.
METHOD.--
1. Prepare four sets of cultivations:
(A) Sloped glucose formate agar, and incubate aerobically at 37° C.
Sloped glucose formate gelatine, and incubate aerobically at 20° C.
(B) Sloped glucose agar to incubate anaerobically at 37° C.
Sloped glucose formate gelatine to incubate anaerobically at 20° C.
(C) Sloped glucose formate agar to incubate anaerobically at 37° C.
Glucose formate bouillon to incubate anaerobically at 37° C.
(D) Sloped glucose formate gelatine to incubate anaerobically at 20° C.
Glucose formate bouillon to incubate anaerobically at 20° C.
2. Seal the cultures forming set B in Buchner's tubes (_vide_ page 239).
3. Seal the cultures forming set C in Bulloch's apparatus; exhaust the
air by means of a vacuum pump, and provide for the absorption of any
residual oxygen by the introduction of pyrogallic acid and caustic soda
in solution (_vide_ page 245). Treat set D in the same way.
4. Observe the cultivations macroscopically and microscopically at
intervals of twenty-four hours until the completion, if necessary, of
seven days' incubation.
5. Control these results.
_Gases Other than Oxygen._--
_Apparatus Required:_
Bulloch's apparatus.
Sterile gas filter (_vide_ page 40).
Gasometer containing the gas it is desired to test (SO_{2}, N_{2}O, NO,
CO_{2}, etc.) or gas generator for its production.
METHOD.--
1. Prepare at least seven tube cultivations upon solid media and deposit
them in Bulloch's apparatus.
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