beyond the possibility of doubt, that we may prove that yeast is
not the only organized ferment able to live and multiply when out
of the influence of free oxygen.
Into a flask, like that represented in FIG. 9, of 2.5 litres
(about four pints) in capacity, we put:
Pure, crystallized, neutral tartrate of lime. .. 100 grammes
Phosphate of ammonia. ... . ... . .. ... . ... 1 grammes
Phosphate of magnesium. ... . ... . ... . ... .. 1 grammes
Phosphate of potassium. ... . ... . ... . .. 0.5 grammes
Sulphate of ammonia. ... . ... . ... . ... .. 0.5 grammes
(1 gramme = 15.43 grains)
To this we added pure distilled water, so as entirely to fill the
flask.
In order to expel all the air dissolved in the water and adhering
to the solid substances, we first placed our flask in a bath of
chloride of calcium in a large cylindrical white iron pot set
over a flame. The exit tube of the flask was plunged in a test
tube of Bohemian glass three-quarters full of distilled water,
and also heated by a flame. We boiled the liquids in the flask
and test-tube for a sufficient time to expel all the air
contained in them. We then withdrew the heat from under the test-
tube, and immediately afterwards covered the water which it
contained with a layer of oil and then permitted the whole
apparatus to cool down.
[Illustration with caption: Fig. 9]
Next day we applied a finger to the open extremity of the exit-
tube, which we then plunged in a vessel of mercury. In this
particular experiment which we are describing, we permitted the
flask to remain in this state for a fort-night. It might have
remained there for a century without ever manifesting the least
sign of fermentation, the fermentation of the tartrate being a
consequence of life, and life after boiling no longer existed in
the flask. When it was evident that the contents of the flask
were perfectly inert, we impregnated them rapidly, as follows:
all the liquid contained in the exit-tube was removed by means of
a fine caoutchouc tube, and replaced by about 1 c. (about 17
minims) of liquid and deposit from another flask, similar to the
one we have just described, but which had been fermenting
spontaneously for twelve days; we lost no time in refilling
completely the exit tube with water which had been first boiled
and then cooled down in carbonic acid gas. This operation lasted
only a few minutes. The exit-tube was again plunged under
mercury. Subsequently the tube was not moved from under the
mercury, and as it formed part of the flask, and there was
neither cork nor india-rubber, any introduction of air was
consequently impossible. The small quantity of air introduced
during the impregnation was insignificant and it might even be
shown that it injured rather than assisted the growth of the
organisms, inasmuch as these consisted of adult individuals which
had lived without air and might be liable to be damaged or even
destroyed by it.
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