Joseph Butler - "Human Nature and Other Sermons"

Louis Antoine Fauvelet de Bourrienne - "Memoirs of Napoleon Bonaparte, v12"

Henry Lindlahr - "Nature Cure"

Edna Ferber - "Roast Beef, Medium"

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Books: The Harvard Classics Volume 38

V >> Various >> The Harvard Classics Volume 38




M. Robin, however, would have no difficulty in determining the
limits of the two kingdoms. According to him, "every variety of
cellulose is, we may say, insoluble in ammonia, as also are the
reproductive elements of plants, whether male or female. Whatever
phase of evolution the elements which reproduce a new individual
may have reached, treatment with this reagent, either cold or
raised to boiling, leaves them absolutely intact under the eyes
of the observer, except that their contents, from being partially
dissolved, become more transparent. Every vegetable whether
microscopic or not, every mycelium and every spore, thus
preserves in its entirety its special characteristics of form,
volume and structural arrangements; whilst in the case of
microscopic animals, or the ova and microscopic embryos of
different members of the animal kingdom, the very opposite is the
case."

We should be glad to learn that the employment of a drop of
ammonia would enable us to pronounce an opinion with this degree
of confidence on the nature of the lowest microscopic beings; but
is M. Robin absolutely correct in his assumptions? That gentleman
himself remarks that spermatozoa, which belong to animal
organisms, are insoluble in ammonia, the effect of which is
merely to make them paler. If a difference of action in certain
reagents, in ammonia, for example, were sufficient to determine
the limits of the animal and vegetable kingdoms, might we not
argue that there must be a very great and natural difference
between moulds and bacteria, inasmuch as the presence of a small
quantity of acid in the nutritive medium facilitates the growth
and propagation of the former, whilst it is able to prevent the
life of bacteria and vibrios? Although as is well known, movement
is not an exclusive characteristic of animals, yet we have always
been inclined to regard vibrios as animals, on account of the
peculiar character of their movements. How greatly they differ in
this respect from the diatomacae, for example! When the vibrio
encounters an obstacle it turns, or after assuring itself by some
visual effort or other that it cannot overcome it, it retraces
its steps. The colpoda--undoubted infusoria--behave in an exactly
similar manner. It is true one may argue that the zoospores of
certain cryptogamia exhibit similar movements; but do not these
zoospores possess as much of an animal nature as do the
spermatozoa? As far as bacteria are concerned, when, as already
remarked, we see them crowd round a bubble of air in a liquid to
prolong their life, oxygen having failed them everywhere else,
how can we avoid believing that they are animated by an instinct
for life, of the same kind that we find in animals? M. Robin
seems to us to be wrong in supposing that it is possible to draw
any absolute line of separation between the animal and vegetable
kingdoms. The settlement of this line however, we repeat again,
no matter what it may be, has no serious bearing upon the
questions that have been the subject of our researches.

In like manner the difficulty which M. Robin has raised in
objecting to the employment of the word GERM, when we cannot
specify whether the nature of that germ is animal or vegetable,
is in many respects an unnecessary one. In all the questions
which we have discussed, whether we were speaking of fermentation
or spontaneous generation, the word GERM has been used in the
sense of ORIGIN OF LIVING ORGANISM. If Liebig, for example, said
of an albuminous substance that it gave birth to ferment, could
we contradict him more plainly than by replying "No; ferment is
an organized being, the germ of which is always present, and the
albuminous substance merely serves by its occurrence to nourish
the germ and its successive generations"?

In our Memoir of 1862, on so-called SPONTANEOUS generations,
would it not have been an entire mistake to have attempted to
assign specific names to the microscopic organisms which we met
with in the course of our observations? Not only would we have
met with extreme difficulty in the attempt, arising from the
state of extreme confusion which even in the present day exists
in the classification and nomenclature of these microscopic
organisms, but we should have been forced to sacrifice clearness
in our work besides; at all events, we should have wandered from
our principal object, which was the determination of the presence
or absence of life in general, and had nothing to do with the
manifestation of a particular kind of life in this or that
species, animal or vegetable. Thus we have systematically
employed the vaguest nomenclature, such as mucors, torulae,
bacteria, and vibrios. There was nothing arbitrary in our doing
this, whereas there is much that is arbitrary in adopting a
definite system of nomenclature, and applying it to organisms but
imperfectly known, the differences or resemblances between which
are only recognizable through certain characteristics, the true
signification of which is obscure. Take, for example, the
extensive array of widely different systems which have been
invented during the last few years for the species of the genera
bacterium and vibrio in the works of Cohn, H. Hoffmann, Hallier,
and Billroth. The confusion which prevails here is very great,
although we do not of course by any means place these different
works on the same footing as regards their respective merits.

M. Robin is, however, right in recognizing the impossibility of
maintaining in the present day, as he formerly did, "That
fermentation is an exterior phenomenon, going on outside
cryptogamic cells, a phenomenon of contact. It is probably," he
adds, "an interior and molecular action at work in the innermost
recesses of the substance of each cell." From the day when we
first proved that it is possible for all organized ferments,
properly so called, to spring up and multiply from their
respective germs, sown, whether consciously or by accident, in a
mineral medium free from organic and nitrogenous matters other
than ammonia, in which medium the fermentable matter alone is
adapted to provide the ferment with whatever carbon enters into
its composition, from that time forward the theories of Liebig,
as well of Berzelius, which M. Robin formerly defended, have had
to give place to others more in harmony with facts. We trust that
the day will come when M. Robin will likewise acknowledge that he
has been in error on the subject of the doctrine of spontaneous
generation, which he continues to affirm, without adducing any
direct proofs in support of it, at the end of the article to
which we have been here replying.

We have devoted the greater part of this chapter to the
establishing with all possible exactness the extremely important
physiological fact of life without air, and its correlation to
the phenomena of fermentations properly so called--that is to
say, of those which are due to the presence of microscopic
cellular organisms. This is the chief basis of the new theory
that we propose for the explanation of these phenomena. The
details into which we have entered were indispensable on account
of the novelty of the subject no less than on account of the
necessity we were under of combating the criticisms of the two
German naturalists, Drs. Oscar Brefeld and Traube, whose works
had cast some doubts on the correctness of the facts upon which
we had based the preceding propositions. We have much pleasure in
adding that at the very moment we were revising the proofs of
this chapter, we received from M. Brefeld an essay, dated Berlin,
January, 1876, in which, after describing his later experimental
researches, he owns with praiseworthy frankness that Dr. Traube
and he were both of them mistaken. Life without air is now a
proposition which he accepts as perfectly demonstrated. He has
witnessed it in the case of Mucor racemosus and has also verified
it in the case of yeast. "If," he says, "after the results of my
previous researches, which I conducted with all possible
exactness, I was inclined to consider Pasteur's assertion as
inaccurate and to attack them, I have no hesitation now in
recognizing them as true, and in proclaiming the service which
Pasteur has rendered to science in being the first to indicate
the exact relation of things in the phenomenon of fermentation."
In his later researches, Dr. Brefeld has adopted the method which
we have long employed for demonstrating the life and
multiplication of butyric vibrios in the entire absence of air,
as well as the method of conducting growths in mineral media
associated with fermentable substance. We need not pause to
consider certain other secondary criticisms of Dr. Brefeld. A
perusal of the present work will, we trust, convince him that
they are based on no surer foundation than were his former
criticisms.

To bring one's self to believe in a truth that has just dawned
upon one is the first step towards progress; to persuade others
is the second. There is a third step, less useful perhaps, but
highly gratifying nevertheless, which is, to convince one's
opponents.

We therefore, have experienced great satisfaction in learning
that we have won over to our ideas an observer of singular
ability, on a subject which is of the utmost importance to the
physiology of cells.




VI. REPLY TO THE CRITICAL OBSERVATIONS OF LIEBIG, PUBLISHED IN
1870.

[Footnote: LIEBIG, Sur la fermentation et la source de la force
musculaire (Annales de Chimie et de Physique, 4th series, t.
xxiii., p. 5, 1870).]

In the Memoir which we published, in 1860, on alcoholic
fermentation, and in several subsequent works, we were led to a
different conclusion on the causes of this very remarkable
phenomenon from that which Liebig had adopted. The opinions of
Mitscherlich and Berzelius had ceased to be tenable in the
presence of the new facts which we had brought to light. From
that time we felt sure that the celebrated chemist of Munich had
adopted our conclusions, from the fact that he remained silent on
this question for a long time, although it had been until then
the constant subject of his study, as is shown by all his works.
Suddenly there appeared in the Annales de Chimie et de Physique a
long essay, reproduced from a lecture delivered by him before the
Academy of Bavaria in 1868 and 1869. In this Liebig again
maintained, not, however, without certain modifications, the
views which he had expressed in his former publications, and
disputed the correctness of the principal facts enunciated in our
Memoir of 1860, on which were based the arguments against his
theory.

"I had admitted," he says, "that the resolution of fermentable
matter into compounds of a simpler kind must be traced to some
process of decomposition taking place in the ferment, and that
the action of this same ferment on the fermentable matter must
continue or cease according to the prolongation or cessation of
the alteration produced in the ferment. The molecular change in
the sugar, would, consequently, be brought about by the
destruction or modification of one or more of the component parts
of the ferment, and could only take place through the contact of
the two substances. M. Pasteur regards fermentation in the
following light: The chemical action of fermentation is
essentially a phenomenon correlative with a vital action,
beginning and ending with it. He believes that alcoholic
fermentation can never occur without the simultaneous occurrence
of organization, development, and multiplication of globules, or
continuous life, carried on from globules already formed. But the
idea that the decomposition of sugar during fermentation is due
to the development of the cellules of the ferment, is in
contradiction with the fact that the ferment is able to bring
about the fermentation of a pure solution of sugar. The greater
part of the ferment is composed of a substance that is rich in
nitrogen and contains sulphur. It contains, moreover, an
appreciable quantity of phosphates, hence it is difficult to
conceive how, in the absence of these elements in a pure solution
of sugar undergoing fermentation, the number of cells is capable
of any increase."

Notwithstanding Liebig's belief to the contrary, the idea that
the decomposition of sugar during fermentation is intimately
connected with a development of the cellules of the ferment, or a
prolongation of the life of cellules already formed, is in no way
opposed to the fact that the ferment is capable of bringing about
the fermentation of a pure solution of sugar. It is manifest to
any one who has studied such fermentation with the microscope,
even in those cases where the sweetened water has been absolutely
pure, that ferment-cells do multiply, the reason being that the
cells carry with them all the food-supplies necessary for the
life of the ferment. They may be observed budding, at least many
of them, and there can be no doubt that those which do not bud
still continue to live; life has other ways of manifesting itself
besides development and cell-proliferation.

If we refer to the figures on page 81 of our Memoir of 1860,
Experiments D, E, F, H, I, we shall see that the weight of yeast,
in the case of the fermentation of a pure solution of sugar,
undergoes a considerable increase, even without taking into
account the fact that the sugared water gains from the yeast
certain soluble parts, since in the experiments just mentioned,
the weights of solid yeast, washed and dried at 100 degrees C.
(212 degrees F.), are much greater than those of the raw yeast
employed, dried at the same temperature.

In these experiments we employed the following weights of yeast,
expressed in grammes (1 gramme=15.43 grains):

(1) 2.313

(2) 2.626

(3) 1.198

(4) 0.699

(5) 0.326

(6) 0.476

which became, after fermentation, we repeat, without taking into
account the matters which the sugared water gained from the
yeast:

grammes. grains.
(1) 2.486 Increase 0.173 = 2.65

(2) 2.963 Increase 0.337 = 5.16

(3) 1.700 Increase 0.502 = 7.7

(4) 0.712 Increase 0.013 = 0.2

(5) 0.325 Increase 0.009 = 0.14

(6) 0.590 Increase 0.114 = 1.75

Have we not in this marked increase in weight a proof of life,
or, to adopt an expression which may be preferred, a proof of a
profound chemical work of nutrition and assimilation?

We may cite on this subject one of our earlier experiments, which
is to be found in the Comptes rendus de l'Academie for the year
1857, and which clearly shows the great influence exerted on
fermentation by the soluble portion that the sugared water takes
up from the globules of ferment:

"We take two equal quantities of fresh yeast that have been
washed very freely. One of these we cause to ferment in water
containing nothing but sugar, and, after removing from the other
all its soluble particles--by boiling it in an excess of water
and then filtering it to separate the globules--we add to the
filtered liquid as much sugar as was used in the first case along
with a mere trace of fresh yeast insufficient, as far as its
weight is concerned, to affect the results of our experiment. The
globules which we have sown bud, the liquid becomes turbid, a
deposit of yeast gradually forms, and, side by side with these
appearances, the decomposition of the sugar is effected, and in
the course of a few hours manifests itself clearly. These results
are such as we might have anticipated. The following fact,
however, is of importance. In effecting by these means the
organization into globules of the soluble part of the yeast that
we used in the second case, we find that a considerable quantity
of sugar is decomposed. The following are the results of our
experiment; 5 grammes of yeast caused the fermentation of 12.9
grammes of sugar in six days, at the end of which time it was
exhausted. The soluble portion of a like quantity of 5 grammes of
the same yeast caused the fermentation of 10 grammes of sugar in
nine days, after which the yeast developed by the sowing was
likewise exhausted."

How is it possible to maintain that, in the fermentation of water
containing nothing but sugar, the soluble portion of the yeast
does not act, either in the production of new globules or the
perfection of old ones, when we see, in the preceding experiment,
that after this nitrogenous and mineral portion has been removed
by boiling, it immediately serves for the production of new
globules, which, under the influence of the sowing of a mere
trace of globules, causes the fermentation of so much sugar?
[Footnote: It is important that we should here remark that, in
the fermentation of pure solution of sugar by means of yeast, the
oxygen originally dissolved in the water, as well as that
appropriated by the globules of yeast in their contact with air,
has a considerable effect on the activity of the fermentation. As
a matter of fact, if we pass a strong current of carbonic acid
through the sugared water and the water in which the yeast has
been treated, the fermentation will be rendered extremely
sluggish, and the few new cells of yeast which form will assume
strange and abnormal aspects. Indeed this might have been
expected, for we have seen that yeast, when somewhat old, is
incapable of development or of causing fermentation even in a
fermentable medium containing all the nutritive principles of
yeast if the liquid has been deprived of air; much more should we
expect this to be the case in pure sugared water, likewise
deprived of air.]

In short, Liebig is not justified in saying that the solution of
pure sugar, caused to ferment by means of yeast, contains none of
the elements needed for the growth of yeast, neither nitrogen,
sulphur nor phosphorus, and that, consequently, it should not be
possible, by our theory, for the sugar to ferment. On the
contrary, the solution does contain all these elements, as a
consequence of the introduction and presence of the yeast.

Let us proceed without examination of Liebig's criticisms:

"To this," he goes on to say, "must be added the decomposing
action which yeast exercises on a great number of substances, and
which resembles that which sugar undergoes. I have shown that
malate of lime ferments readily enough through the action of
yeast, and that it splits up into three other calcareous salts,
namely, the acetate, the carbonate and the succinate. If the
action of yeast consists in its increase and multiplication, it
is difficult to conceive this action in the case of malate of
lime and other calcareous salts of vegetable acids."

This statement, with all due deference to the opinion of our
illustrious critic, is by no means correct. Yeast has no action
on malate of lime, or on other calcareous salts formed by
vegetable acids. Liebig had previously, much to his own
satisfaction, brought forward urea as being capable of
transformation into carbonate of ammonia during alcoholic
fermentation in contact with yeast. This has been proved to be
erroneous. It is an error of the same kind that Liebig again
brings forward here. In the fermentation of which he speaks (that
of malate of lime), certain spontaneous ferments are produced,
the germs of which are associated with the yeast, and develop in
the mixture of yeast and malate. The yeast merely serves as a
source of food for these new ferments without taking any direct
part in the fermentations of which we are speaking. Our
researches leave no doubt on this point, as is evident from the
observations on the fermentation of tartrate of lime previously
given.

It is true that there are circumstances under which yeast brings
about modifications in different substances. Doebereiner and,
Mitscherlich, more especially, have shown that yeast imparts to
water a soluble material, which liquefies cane-sugar and produces
inversion in it by causing it to take up the elements of water,
just as diastase behaves to starch or emulsin to amygdalin.

M. Berthelot also has shown that this substance may be isolated
by precipitating it with alcohol, in the same way as diastase is
precipitated from its solutions. [Footnote: DOEBEREINER, Journal
de Chimie de Schweigger, vol. xii., p. 129, and Journal de
Pharmacie, vol. i., p. 342.

MITSCHERLICH, Monatsberichte d. Kon. Preuss. Akad. d. Wissen, eu
Berlin, and Rapports annuels da Berzelius, Paris, 1843, 3rd year.
On the occasion of a communication on the inversion of cane-sugar
by H. Rose, published in 1840, M. Mitscherlich observed: "The
inversion of cane-sugar in alcoholic fermentation is not due to
the globules of yeast, but to a soluble matter in the water with
which they mix. The liquid obtained by straining off the ferment
on a filter paper possesses the property of converting cane-sugar
into uncrystallizable sugar."

BERTHELOT, Comptes rendus de l'Academie. Meeting of May 28th,
1860, M. Berthelot confirms the preceding experiment of
Mitscherlich, and proves, moreover, that the soluble matter of
which the author speaks may be precipitated with alcohol without
losing its invertive power.

M. Bechamp has applied Mitscherlich's observation, concerning the
soluble fermentative part of yeast, to fungoid growths, and has
made the interesting discovery that fungoid growths, like yeast,
yield to water a substance that inverts sugar. When the
production of fungoid growths is prevented by means of an
antiseptic, the inversion of sugar does not take place.

We may here say a few words respecting M. Bechamp's claim to
priority of discovery. It is a well-known fact that we were the
first to demonstrate that living ferments might be completely
developed if their germs were placed in pure water together with
sugar, ammonia, and phosphates. Relying on this established fact,
that moulds are capable of development in sweetened water in
which, according to M. Bechamp, they invert the sugar, our author
asserts that he has proved that "living organized ferments may
originate in media which contain no albuminous substances." (See
Comptes rendus, vol. ixxv., p. 1519.) To be logical, M. Bechamp
might say that he has proved that certain moulds originate in
pure sweetened water without nitrogen or phosphates or other
mineral elements, for such a deduction might very well be drawn
from his work, in which we do not find the least expression of
astonishment at the possibility of moulds developing in pure
water containing nothing but sugar without other mineral or
organic principles.

M. Bechamp's first note on the inversion of sugar was published
in 1855. In it we find nothing relating to the influence of
moulds. His second, in which that influence is noticed, was
published in January, 1858, that is, subsequently to our work on
lactic fermentation, which appeared in November, 1857. In that
work we established for the first time that the lactic ferment is
a living, organized being, that albuminous substances have no
share in the production of fermentation, and that they only serve
as the food of the ferment. M. Bechamp's note was even subsequent
to our first work on alcoholic fermentation, which appeared on
December 21st, 1857. It is since the appearance of these two
works of ours that the preponderating influence of the life of
microscopic organism in the phenomena of fermentation has been
better understood. Immediately after their appearance M. Bechamp,
who from 1855 had made no observation on the action of fungoid
growths on sugar, although he had remarked their presence,
modified his former conclusions. (Comptes rendus, January 4th,
1858.)] These are remarkable facts, which are, however, at
present but vaguely connected with the alcoholic fermentation of
sugar by means of yeast. The researches in which we have proved
the existence of special forms of living ferments in many
fermentations, which one might have supposed to have been
produced by simple contact action, had established beyond doubt
the existence of profound differences between those
fermentations, which we have distinguished as fermentations
proper, and the phenomena connected with soluble substances. The
more we advance, the more clearly we are able to detect these
differences. M. Dumas has insisted on the fact that the ferments
of fermentation proper multiply and reproduce themselves in the
process whilst the others are destroyed. [Footnote: "There are
two classes of ferments; the first, of which the yeast of beer
may be taken as the type, perpetuate and renew themselves if they
can find in the liquid in which they produce fermentation food
enough for their wants; the second, of which diastase is the
type, always sacrifice themselves in the exercise of their
activity." (DUMAS, Comptes rendus de l'Academie, t. lxxv., p.
277, 1872.)] Still more recently M. Muntz has shown that
chloroform prevents fermentations proper, but does not interfere
with the action of diastase (Comptes rendus, 1875). M. Bouchardat
had already established the fact that hydrocyanic acid, salts of
mercury, ether, alcohol, creosote, and the oils of turpentine,
lemon, cloves, and mustard destroy or check alcoholic
fermentations, whilst in no way interfering with the glucoside
fermentations (Annales de Chimie et de Physique. 3rd series, t.
xiv., 1845). We may add in praise of M. Bouchardat's sagacity,
that that skilful observer has always considered these results as
a proof that alcoholic fermentation is dependent on the life of
the yeast-cell, and that a distinction should be made between the
two orders of fermentation.