Yogurt Recipe

One of the first things people ask me when I tell them I make my own dairy products is do I make yogurt. Actually yogurt was the first thing I learned how to make. Many years ago I purchased an electric yogurt maker and proceeded to make the delightful dairy product using powdered milk. But now that I have plenty of goat milk I've experimented with making yogurt as simply as possible without the use of special equipment.

Yogurt is the Turkish word for milk that has been curdled with a lactic starter. Many countries have a long tradition of cooking with and eating some form of lactic cultured milk. Yogurt is easily digested and it keeps better than milk, which is why it's popular in tropical climes.

Yogurt is formed by the growth of two bacterial organisms in milk; Streptococcus thermophilus* (a warmth loving bacteria) and Lactobacillus bulgaricus (a strain of bacteria from Bulgaria, where we all know they make great yogurt) which turn the milk sugars into lactic acid. Lactic acid is much easier for your body to digest than milk sugars, so even people who can't drink milk (lactose intolerant) can still "handle" yogurt. Oft times you will also find yogurt that contains other "Probiotic" cultures such as Lactobacillus acidophilus, Bifidobacterium longum, and Bifidobacterium infantis which are bacterium normally found in your intestines. Together these bacteria aid in digestion and the synthesis of vitamins. If you are taking antibiotics, which tend to kill your "intestinal flora" as well as whatever is making you ill, you should eat plenty of yogurt to help replace the "good guys" in your digestive system. Continue Reading >>

The Fairy Tale of E-coli Causing Sickness and Death

This was an email sent to me today concerning E-coli from spinach causing sickness or even death. It came as a result, I am quite sure, of the AOL news coverage and the hour-long CNN documentary which has been running all weekend about the spinach scare in 2006, farming practices, the problems of the FDA, the wrongly theorized cause, the supposed remedies, and lawsuits and so on.

It also highlighted the death of an elderly woman and the near-death of a little girl. I am always a bit suspicious and fearful of news media, like politicians, who spend too much time telling me what to be afraid of and who to blame." This is especially true when most of the accompanying ads seem to be from the pharmaceutical industry.

I am sharing with you my thoughts and response to questions. The host and interviewer of the piece was Dr. Sanjay Gupta, CNN's medical correspondent.

Here is the email letter:
"Dear Dr. Young "I was just wondering if the "germ" finds a friendly environment (over acidic), can it multiply and create more waste and therefore there is a possibility of exposure that leaves an acidic body in jeopardy.

"Obviously E-coli has been used for ages in classrooms (because it doesn't take much to provide it with the necessary ingredients for quick replication for observation) without hurting anyone, but if for some reason, some crazy kid decided to lick his slide and the internal terrain was compromised already, would the e-coli (or any other bacteria) then be able to replicate internally and produce enough waste to tip the balance (though I would have thought it would take ages)? Or is that really out of the question? Clearly the focus is bas-ackwards (as they say) in western etiology.

"When a group of kids who have been mistakenly identified as being "struck" by E-coli from a bad meat source, is it simply that the "'bad" (is there any good? : ) meat has become so acidic that it is like drinking industrial solvents? Would chemical tests of the suspect sources reveal the problem? Biological interpretations appear to be just wrong.
"Thank you so much for taking the time to answer these!"
Sally

Dear Sally: Thank you for your email questions. I have stated in my writings that germs (the "germing" process) cannot cause disease but "germs" themselves do produce digestive (enzymes) or metabolic waste products called exotoxins and mycotoxins or simply acids that contribute to disease states.

I have further stated that there is only one cause of sickness, dis-ease and death and that is the over acidification of the tissues (latent tissue acidosis) and then blood (compensated acidosis and then decompensated acidosis) due to an inverted way of living, eating and thinking.

Your question of whether or not bacteria can cause disease is an important question that has been debated for over 100 years. As you know, the French scientist, Antione BeChamp, an adversary of Louis Pasteur, said, "the germ is nothing; the terrain is everything."

Maintaining the healthy alkaline environment or terrain of the human body is critical for good health and for the prevention of any disease. The human body is alkaline by design and acidic by function. That is, the body is designed to run on primarily alkaline fuel because all bodily functions create acids which must be largely neutralized to protect the body itself from the acidic creation of disease and dis-ease.

The body maintains this alkaline pH design at 7.365 by eliminating gastrointestinal and metabolic acids through urination, perspiration, defecation and respiration. When we eat anything, including highly alkaline spinach, there will be residues of acids that can be easily buffered from the sodium bicarbonate produced and released in the stomach. So what comes first the bacteria or the acid? What we have here is the chicken or the egg scenario. I would suggest to you that everything is the prey of life and nothing is the prey of death. What can be nourished can be consumed and everything is simply trying to live.

Since our bodies run on energy in the form of electrons, the by-products of energy consumption is always acid. When acids from the process of metabolism are not properly eliminated, they can spoil our cells that make up our tissues that then gives rise to biological transformations known as bacteria.

So the answer to the question, "what comes first the bacteria or the acid?" the answer is clearly the ACID. Acid is the bi-product of energy being used or consumed. Bacteria is then a by-product of energy being consumed like the smoke from a fired gun, of spoiling or degenerating matter not the cause of the spoiling or degenerating matter. ACID is the only cause for spoiling of degenerating matter or tissue! In today's headline on AOL News it said, "Two More Deaths Possibly Linked to Tainted Spinach." A key word in this headline is "possibly" which infers that scientific investigators just don't know! And I believe that they know that they don't know. But they are going to try and calm your fears--and so they are going to come up with some sort of explanation that they can sell to the public.

But I am convinced that tainted or fermenting spinach which would contain very little amounts of oxalic acid and not enough to make one sick and it would ordinarily be neutralized by the sodium bicarbonate secreted in the mouth, stomach and intestines. Why? Because if spinach is that tainted or fermenting it would have a terrible smell, a terrible taste, and unlikely that anyone in their right mind would eat it. States of ultimate sickness, disease and then death comes as a process of poor lifestyle and dietary choice that then lead to a state of over acidity. The oxalic acid from a few leaves of Spinach could not possibly shut down the kidneys.
Now consider this: Most people get their spinach from sealed plastic bags designed to keep the spinach fresh for awhile. My bet would be that of the 250 or so spinach leaves in that one bag, NO SINGLE LEAF came from even the same plant. By the time the leaves are separated from the plant, washed and rewashed, tumbled and tossed, run over the multiple conveyer belts, those leaves came from all over the farm field, and sometimes from different truck loads and possibly even different fields--all in the same bag at your grocery store.

Have you ever seen the size of those commercial spinach fields? They look like a square mile or bigger. And there is field after field after field. Perhaps we are sophisticated enough to track a bag from the consumer, back to the store, back to the packager, and back to the commercial farmer, and maybe even back to one or two farm fields. But the chance that the FDA or any governmental agencies is sophisticated enough to isolate e-coli down to a few plants or area of a field is beyond my comprehension and certainly my confidence in the U.S. government.
In that enormous field, whatever they found, it is my guess that they would have found approximately the same thing anywhere in the field. And they could go next farm over and find the same results. Thousands of bacteria are everywhere as a product of evolution and a stage of life transforming.

You say the spinach field was too close to a field of cattle? Have you ever driven through California or most other states and looked at the farms? There are thousands of cattle fields and hog farms in proximity to fields of trees, plants and vegetables. There's e-coli everywhere. You have had plenty of e-coli come and go in your body. Fifteen million e-coli bacteria can sit on the head of a pin. I also believe that a hundred thousand people ate spinach leaves from the same field, and if there was some E-coli present, thousands and thousands of people ate those leaves and did not get sick.

What I do know and what I do believe is that many people could conceivable had perhaps some miniscule amount of bacteria, as we all do, from whatever source, and yet, the blood cells, tissues, and rivers/fluids of these peoples' bodies were acidic to begin with.

This internal-external distinction is important because it helps us in the treatment of the dis-ease as we change our focus from the bacteria to the state of over-acidity pH versus the actual alkaline pH of the fluids of the body. And if someone is taken to the hospital with whatever symptoms, they are then treated with a myriad of acidic components, including antibiotics, adding fuel to a fire already started. Rather than using acidic drugs to kill some harmless bacteria, the focus should change to reestablishing the alkaline internal pH environment with alkaline buffers such as sodium bicarbonate.

But hospitals do not do that because they are operating from the same wrong-headed theory as are many governments of the world, U.S. health agencies, medical schools, research laboratories, and lastly the 1,000 pound pharmaceutical gorillas that--along with your tax dollars--fund the whole she-bang.

To suggest that those individuals died from E-coli found in the kidney is like blaming the smoke from a fired gun as the cause of death. Logically we know that smoke from a fired gun cannot kill. We can even argue that it is not the bullet that can kill. It is the person that is pulling the trigger that causes the gun to be fired that causes the release of the bullet and then the residue of smoke. E-coli is the smoke. The bullet is the acid. And the triggering factor is the individual's lifestyle and dietary choices.

Not for a moment do I believe that anyone, unless staving to death, would eat spoiled fermenting smelly and awful tasting spinach. I do not believe that people are that silly, and I am not about to believe the ridiculous claims that spinach was the possible cause of death as suggested by western germ theory scientists. To justify their claims, these medical savants are now saying that E-coli is the possible villain in this fairy tale by suggesting that it is coming from migrant workers who are urinating and defecating in the fields around the spinach, or the nearby cattle and their excrement, or the pig farm, or the water run off that soaked an area of the field.

I can hardly contain myself from laughing out loud when organic farmers are using chicken excrement to fertilize the fields of spinach and other vegetables and fruits, and it certainly is not all "treated" fertilizer. Here at Rancho del Sol, we have used chicken excrement in and around our organic grapefruit and avocado trees for its oxygen and nitrogen components. This is what all organic farmers use. So what would be worse, human excrement or chicken excrement used to fertilize? The point is, we must focus on the cause not the effect and the cause will always be where you find the poison, or the acid, not the "germ" or the "germing process". As for Legionnaire's disease this also is not caused by bacteria. It is caused from over indulging in acidic foods and drinks. And plenty ate the same food and didn't get sick. And there are plenty of those who were sick that had been diagnosed with Legionnaire's where no bacteria could be found. The reason? Acid makes us sick not bacteria.

This is also the case with individuals diagnosed with HIV/AIDS. They are sick but there is no virus present! So what is the cause? It can only be from an over acidic environment. So, we need to look at the acid from our lifestyle and dietary choices.

The acid from the beverages we drink such as tea, coffee and alcohol. The toxins from meats that release nitric, uric, sulfuric, and phosphoric acids. Or the toxins from sugar like acetlyaldehyde or lactic acids. These are the true culprits or poisons that make us sick, tired and fat that lead to our eventual death. Finally you asked the following questions: The first was...I was just wondering that when the germ finds a friendly environment (over acidic), can it multiply and create more waste and therefore is there a possibility of exposure that leaves an acidic body in jeopardy. The word "germ" comes from the German language which means to sprout or germinate. Allow me to digress a moment. Germs are not really nouns, even though we think of them as "things" but they are not; "germ" should be a verb--or a noun derived from a verb. I think it should be a gerund if I remember my English correctly. A word that ends in "ing." It's not a thing so much as it's an activity. It should be called "germing." Instead we say germination and germinating.

"Germs" are the germinating function of changing matter and are NOT species specific, that is, they do note mate or reproduce. The reality is that germs are not things but actions or reactions from a changing environment. The germ or germination or germing is the expression of that change in matter and should never be classified. Simply, E-coli is a stage of transforming matter and not a reproduction due to an over acidic environment.

E-coli is the change of matter or tissue in a changed environment that is pH sensitive. It is no different than taking water, a liquid and seeing the change that takes place when we change the temperature to zero degrees Celsius and the water changes to ice, a solid. It is still water but in a different form. And so it is with E-coli. E-coli is a form of matter that is born out of the cell when the pH of the environment becomes acidic.

The second question was...obviously E-coli has been used for ages in class rooms (because it doesn't take much to provide it with the necessary ingredients for quick replication for observation) without hurting anyone, but if for some reason some crazy kid decided to lick his slide and the internal terrain was compromised already, would the e-coli (or any other bacteria) then be able to replicate internally and produce enough waste to tip the balance (though I would have thought it would take ages)? Or is that really out of the question? Clearly the focus is bas-ackwards (as they say) in western etiology? Once again germs cannot cause disease even if the child licks the slide. This experiment was done many years ago when Claude Bernard a French physiologist drank a glass of cholera bacteria with little affect other than some nausea. In fact, to prove my point I am willing to eat E-coli on fresh spinach leaves for CNN if they are willing and ready for the TRUTH! Another question was....when a group of kids who have been mistakenly identified as being "struck" by E-coli from a bad meat source, is it simply that the "bad" meat has become so acidic that it is like drinking industrial solvents? Industrial solvents differ greatly, but yes, depending upon some variables, eating any meat is highly acidic and would be like drinking industrial solvent, assuming their quantities and other factors were similar. Another question was....would chemical tests of the suspect sources reveal the problem?

Yes, if tested you would find an increase of specific acids in the suspected sources. Question....are biological interpretations....just wrong? You are right in your suspicions. Biological interpretation is wrong because scientists are focused on the matter rather than focused on the environment around the matter. It comes back to the fish bowl metaphor. When the fish is sick, do you treat the fish or change the water? Western medical science is focused on the fish, treats the fish and then neglects to clean up the environment not realizing that the fish is only as healthy as the environment it is swimming in.

This is true with the fluids of our body. If we find E-coli in the tissues, this is a transformation of the tissues due to the acidic fluids found in and around that tissue. There is no infection, only an outfection of matter giving birth to bacteria due to fluid acidosis. The key to staying healthy is to eat fresh organic greens whenever possible, to build healthy blood, and to help maintain the alkaline pH design of our body.

That includes eating lots of fresh organic spinach! Stay away from the acidic foods, liquids, supplements, and treatments. And especially stay away from those faulty or "acidic" theories of western scientific thought that are based upon a false belief that "germs" cause disease -- it could kill you. This fairy tale of E-coli causing sickness and death is just over the top, it smells of big money, and I see lots of dubious irradiation cost and solutions just down the road. Kindest Regards, Robert O. Young Ph.D.

CHAPTER 1.: The Blood and the Third Anatomical Element by Antoine Bechamp

OF THE NATURE OF FIBRIN ISOLATED FROM THE CLOT OR OBTAINED BY WHIPPING THE BLOOD. THE BLOOD FIBRIN. FIBRINOUS MICRO-ZYMAS. FIBRIN AND OXYGENATED WATER. THE FERMENT OF FIBRIN.

Gay-Lussac and Thenard analyzed fibrin as they had analyzed albumen, caseine and gelatine. Thenard said that fibrin was an isolated animal matter; Chevreul said that it was an animal proximate principle and was greatly surprised, after he had discovered oxygenated water, to find that fibrin decomposed it and disengaged the oxygen, as did organic tissues; as, for example, the liver, etc. He even thought that fibrin was the only proximate principle of its kind endowed with this property.1
This fact in the history of fibrin is important; first, because it is the pivot on which turns the demonstration that this substance, reputed a proximate principle, is of the same order as the substance of the bodies which Chevreul called organic bodies; secondly, because, although neglected by physiologists and chemists, it enabled me to place beyond doubt the existence of a third anatomical element of the blood.
I did not set out with the idea of proving that fibrin is a substance of the same order as the organic tissues. Like everybody else, I regarded it as a proximate principle; I had even maintained its specificity against the chemists who contended that it was only coagulated albumen.2
Preliminary steps to the discovery of the real nature of fibrin and of the third anatomical element of the blood. The ancients regarded it as a positive fact that all animal or vege­table matter was spontaneously alterable while putrefying or
1. Thenard, "Traite elementaire de chimie," Vol. I., p. 528. 6th Ed. 1834.2. See Memoir, Essay sur les substances albuminoid. Theses de la Facuhe de Medecir.e de Strasbourg. 1856.
fermenting. In the last century (i.e., the 18th) the chemist, Macquer, established the conditions for these changes; the presence of water, the contact of air and a certain amount of heat. Long after, when in 1837 Cagniard de la Tour regarded beer-yeast as being organized and living, and fermentation as an effect of vegetation, Schwann, generalizing the new conception, endeavoured to show that no organic matter was spontaneously alterable; that the alteration had for cause the presence of organized living things, microscopic cryptogams, vibrioniens; that is to say, ferments, the origin of which, reviving the old hypothesis of Spallanzanil, he ascribed to the germs of the air.
But, notwithstanding many important verifications, the opinion of Schwann did not prevail; the presence of living products in matter undergoing change was conceded, but while some maintained that the alteration preceded the appearance of the organized products, whatever might be their origin, others, admitting the theory of Cagniard, insisted that the living things, the ferments, were the fruit of spontaneous generation.
Schwann's point of view and the hypothesis of germs of the air were so completely abandoned that in 1854 it was admitted as a fact that even cane sugar in watery solution altered spontaneously at the ordinary temperature of the air, becoming what was called invert sugar, grape sugar. Was that true? The inversion of cane sugar, the result of a chemical reaction of reduction by hydration which was produced, as Biot had observed, under the influence of strong acids could it be effected by water only, at ordinary temperature with the aid of lime alone? I wanted to know what to believe, and I instituted experiments which commenced in 1854 and were continued until 1857. Several consequences of the greatest importance resulted and, among others, the first experimental verification of the hypothesis of germs of the air which Schwann, following Spallanzanil had invoked against spontaneous generation. In short I demonstrated:
1st. That a watery solution of cane sugar remains unaltered indefinitely, at ordinary temperature, under the two following conditions: (A) Absolutely protected from access of air and (B) in contact with a limited volume of air, to which have been added certain salts or a suitable (small) quantity of creosote; as, for instance, one to two drops per 100 c.c.
2nd. That the same solution, pure, or with the addition of certain other salts in contact with the same volume of air, permitted the appearance of cryptogamic products, moulds, etc., at the same time that the inversion of the sugar was effected.
3rd. That the moulds are actually the agents, the ferments of the inversion, by secreting the necessary zymas or soluble ferment.
4th. That creosote, which hinders the birth of the moulds, etc., does not prevent developed moulds from effecting the inversion.
And as it is evident that the water and sugar of the solution cannot of themselves give birth to those cryptogamic productions which invert the cane sugar, nor to anything whatever organized and living, the conclusion is inevitable that these experiments verified the hypothesis of the existence of germs in the air.1
1. Annales de Chimie at de physique, 3d S. op. Vol. L1V. p. 28. (1858.)
Cane sugar being a proximate principle, the experiment constituted also the first demonstration that organic matters exist which are unalterable under the conditions specified by Macquer.
To be applicable generally, it was necessary to prove that what was true for cane sugar was also true for any proximate principle, even for albumin, supposed to be so readily alterable that Colin believed it could spontaneously become an alcoholic ferment.1 But there are solutions of proximate principles, even of their mixtures, which contain some albuminoid substance like the solutions of cane sugar; these solutions, with a very small quantity of creosote added, are preserved, although in contact with a limited quantity of air, so that nothing organized makes its appearance, no ferment­ation, no putrefaction takes place. But if among the materials of the mixture there are some which are directly oxydizable by the oxygen of the air, creosote will not hinder the oxydation.
1. In 1858, M. Pasteur believed so little in the existence of germs of ferments in the air, that he asserted that the lactic ferment and beer-yeast were born spontaneously of the albuminoid matter of the fermentable media.
Let us bear in mind this capital fact which has been experimentally verified in every imaginable case: the solutions of isolated proximate principles, or mixtures of them, even albuminoid ones, first creosoted with a suitable (small) dose, exposed to the contact of a limited quantity of ordinary air, allow nothing living to appear, and remain unaltered except in cases where the mixture contains some directly oxydizable principle. In these kinds of experiments the creosote acts either by rendering the medium sterile for the germs or directly upon them, so as to prevent their development.
Organic matters reduced to proximate principles are unalterable under the conditions specified by Macquer, when the influence of the germs of the air is prevented by creosote; then they are so naturally. But Macquer did not take into account proximate principles, of which he had no idea. He really referred only to natural vegetable and animal matters, that which Thenard calls organic tissues and Chevreul organic bodies.
But among the animal matters on which Macquer experimented was milk, which he regarded as an animal emulsion, and held to be alterable of itself. Much later, Donne (an expert micrographist) and most chemists regarded milk as a solution of milk sugar, caseine and of mineral salts, holding an emulsion of butter in solution. Everybody then thought that milk was a pure mixture of proximate principles.a Such a mixture properly creosoted and in contact with a limited volume of air ought to remain unaltered indefinitely. But it was found to be otherwise. Cow's milk sufficiently creosoted at the time of milking, preserved from contact with air or in contact with only a limited amount of air, neither sours nor clots in the ordinary way. The creosote only delays the souring and the consecutive formation of the clot. But it was found that at the moment the milk became clotted, even when the phenomenon takes place in full contact with air, and with or without the addition of creosote, none of the cryptogamic productions could be found which Schwann's experiments led one to look for in it.
[aEvan Landois in his Physiology (Eng.; trans, by Stirling, 1889), makes this mistake. He makes no reference to its ferment.— Trans.]
But the souring and the clot, I do not speak of the coagulation of the milk, is only the first phase in the phenomenon of alteration. The second phase, in spite of the addition of creosote, was characterized by the constant appearance of vibrios or of bacteria. Milk then does not act as would a simple mixture of proximate principles.
These experiments and observations, which date from before 1858, were not published until 1873.1 They had greatly surprised me. Milk then was not what it had been supposed to be. There exist in it organic matters alterable, without the aid of germs of the air, and Macquer was justified in declaring them to be spontaneously alterable. And since, notwithstanding the creosote the milk already altered, soured and
1. C. R., Vol. LXXVI, p. 654.
clotted, permitted the appearance of vibrios in its substance, if these vibrios were not the products of spontaneous generation, to what did they owe their birth? It was experiments contemporary with those upon the calcareous rocks which will be studied in the last chapter of this work, as well as those which led to the discovery of the new category of living organized productions to which I have given the name of microzymas, because of their functions as ferments and of their extreme minuteness.1 It is then the microzymas of the milk itself which are the agents of its alteration and which subsequently becomes vibrios by evolution.
1. C. R., Vol. LXIII, p. 451 (1866).
The method which led to these results, as important as unexpected, the close relation between the geological ferments and the anatomical and physiological ferments of present living animals, and which at the same time answered in the negative the question of the spontaneous generation of organized ferments, is the same which has permitted the demonstration of the inherent unalterability of proximate principles, and to verify the old hypothesis of germs of the air which had been neglected. Thanks to it, it has been possible to explain anatomically and physiologically the phenomena of coagulation and the other spontaneous changes of the blood.
This method had its origin in experiments on the inversion of cane sugar, supposed to be spontaneous, and in those relating to the changes which occurred in milk, which made conspicuous the principle obtained by experiment, that creosote absolutely prevents the alteration of immediate principles by preventing the development of all living organized products, even in contact with a limited quantity of ordinary air, while the same doses under the same condi­tions did not prevent change in natural animal matters, tissues and humors, even permitting them to give birth to vibrios or to bacteria.
It is important to bear in mind that the new method (of experiment) enabled us to distinguish organic matters composed only of proximate principles, from natural vegetable and animal matters, that is to say, from organic bodies properly so-called; in short, to distinguish organic matter in the chemical meaning from that which, like milk,is organic matter in the anatomical and physiological meaning; the ferments which change the former, the organic matters in a chemical sense, that is to say, proximate principles, have for origin the germs of the air, while the ferments which change the second, that is to say, the natural organic matters, are the microzymas of their own substance, which are inherent in them as anatomical elements.
In fact, the phenomenon of the birth of the vibrios in the spontaneously altered milk, if indeed they were the result of the evolution of the microzymas of the milk, ought not to be an isolated fact, but a particular case of a general phenomenon, proper to all organic bodies, so that the fact of the birth of vibrioniens in an organic body, humor or tissue should be considered as evidence of the existence of microzymas in this tissue or this humor, even though the microscope has failed to reveal them.
Experiment has confirmed in every sense these consequences of the application of the new method of investigation to the study of the phenomenon of the spontaneous change of milk. The matter of all the tissues and humors, of all organic bodies, from the highest to the lowest—as, for instance, beer-yeast and the mother of vinegar—may give birth to vibrioniens in like conditions to those in which they are produced in the milk, or in which such conditions can be realized, if it be necessary to encourage otherwise the evolu­tion of their microzymas. And when the phenomenon of spontaneous alteration of such matter is recorded, the matter being protected from germs of the air, without the appearance of vibrioniens, it may be confidently affirmed that microzymas were present and were the agents of the change.
The following is the application of the method to fibrin, regarded as an organic body:
Demonstration that fibrin is not a proximate principle, but a false membrane of microzymas. Birth of bacteria in the fibrin.
The fibrin is produced mechanically from the blood by whipping the latter; being regarded as an organic body, it ought like milk to contain microzymas capable of undergoing vibrionian evolution.
To demonstrate this, M. Estor and I employed a modification of the method which had been used to demonstrate the microzymas of the chalk and of muscle flesh. The modification consisted in preparing a starch of the fecula of potatoes, to boil it for a long time, to creosote it while boiling and to introduce into it the solid substance to be studied at the moment it was extracted from the creosoted water, into which it had been immersed to protect it from the influence of germs of the air. The experiment was as follows: Fibrin was obtained by whipping under the following conditions; at the moment of the venesection creosote was added to the blood, and at the same time it was whipped with a bundle of metallic wires which had just been washed in boiling creo­soted water; then the fibrin was washed in creosoted water. Into 100 grammes of creosoted starch 15 grammes of freshly prepared humid fibrin were introduced and the flask sealed and placed in the oven heated to from 30° to 40° C. (= 86° —104° F.). The starch, exactly as happened with muscle tissue, became liquefied by degrees, and after a time the presence of bacteria in the mixture was evident; but it was observed that the liquifaction of the starch generally preceded the appearance of the bacteria.
The foregoing is a general view of the phenomena, but differences were observed in its manifestations, according to the species and age of the animal as well as the source of the blood. Generally the fibrin of young animals disintegrates in fluid starch, while the bacteria develop. The duration of the liquifaction of the starch is also variable.
It is known that boiled milk clots, which means that the microzymas are not killed at the temperature of boiling; on the other hand, to prevent the chalk from liquifying starch,a I was obliged to heat it (moist) to more than 200° C. (=392° F.). The microzymas of the fibrin resist up to 100° C. (= 212° F.). The fibrin was boiled for several minutes in distilled water before it was placed in the starch. In this case the liquifaction is still further delayed and even ceases to be produced if the boiling of the fibrin is too prolonged; but the bacteria appear none the less; and these bacteria present always the same morphological characters.
[aFor explanation of the action of the microzymas of the calcareous rocks see "Role de la craie dans les fermentations," Bull. Soc. Ch., Vol. VI, p. 484 (1866); also "Les Microzymas," third conference; also post-chapter VIII of this work.—Trans.)
To complete the demonstration it should be added, and M. Estor was witness of the fact, that fibrin, exactly as was the case with the mother of vinegar (a sort of vegetable membrane of visible microzymas), and under like conditions can produce lactic and butyric fermentation, a fact which will be further considered hereafter.
Such was the experiment and its complement, whence it was concluded that fibrin, like milk, like flesh, like the tissue of liver, etc., contains microzymas, since, like them, it gives birth to bacteria without the aid of germs of the air.
Under the conditions of the experiment these micro­zymas could only have come from the blood. Efforts were then made to find them in the blood itself at the moment of the venesection. This was a delicate investigation and will be reverted to hereafter, as it is allied to the whole subject of this work.
Fibrin, whether obtained by whipping or by washing the clot (we will see presently wherein these two preparations differ), is not a proximate principle, but constitutes a membrane or fibre composed of microzymas. In short, it is not organic matter in the chemical meaning, but an organic body in the anatomical and physiological meaning. Nevertheless this demonstration that fibrin contains microzymas is indirect; and one might still contend that the bacteria has been born spontaneously, in the mixture of fibrin with starch. In any case it left unsettled the nature of the substance which, in the false membrane, is like an intermicrozymian gangue, as also the quantitative relation between the microzymas and this substance. It was therefore very desir­able to obtain these microzymas isolated, as Estor and I had isolated those of the liver.
The fibrinous microzymas and their properties compared with those of the fibrin.
The question of the solubility of fibrin in dilute hydrochloric acid had been long discussed and it occurred to me that I might there find a means whereby the microzymas might be isolated, as they ought to be insoluble in it, as were those of the chalk. On going over the history of experiments on the fibrin, I found many experiments and observations relating to fibrin deserving attention which had been neglected.
Thenard had already described the action of dilute hydrochloric acid upon the fibrin of the blood and the formation of hydrochlorates of this substance, one of which, the gelatinous, was soluble in tepid water.1 Long afterwards, Bouchardat called attention to the fact that Chevreul had
1. Thenard, Traiee elementaire de chimie, Vol. III, p. 430 (1815).
demonstrated that fibrin always contained fat and he asked himself if even deprived of fatty bodies it would be a pure proximate principle? To demonstrate that that which had been generally admitted was not well founded, he made the following experiment:
He heated fresh, moist fibrin with ten times its weight of very dilute hydrochloric acid (1 to 2000) and observed that it swelled up, and by a prolonged maceration was at last dissolved, but that there always remained manifest a portion of a product which was not attacked by an excess of this very dilute acid employed as a solvent. Bouchardat called the undissolved part epidermose, and albuminose that which was dissolved.1 It was from this experiment that Bouchardat justly concluded that the fibrin is decidedly not a proximate principle.
1. C.R., Vol. XIV, p. 962 (1842).
The part of the fibrin that Bouchardat regarded under the name of epidermose, as one of the two proximate principles constituting fibrin, was precisely the microzymas which I proposed to isolate. In consequence of the viscosity of the hydrochloric solution, essentially capable of change as Bouchardat had said, and of the slowness of filtration, he had not determined the quantity of the epidermose. By employing a less diluted hydrochloric acid (1 to 3 c.c. of fuming acid per 1000 c.c.) the viscosity was diminished, and on adding to it 2-3 drops of phenol per 100 c.c. the suspected alteration was absolutely prevented, and the deposition of the insoluble part could be awaited. Even then it requires from ten to twelve days to filter a litre of the liquid. For quantitative experiments it must be left at rest and the deposit must not be turned out upon the filter until the filtration of the supernatant liquid is completed.
The grayish brown mass retained on the filter was resolved under the microscope into exceedingly fine molecular granulations, which are the microzymas, and some shapeless fragments proceeding doubtless from the blood globules destroyed during the preparation of the fibrin. To procure these molecular granulations as free as possible from foreign fragments the mass when removed from the filter is steeped in hydrochloric acid (1 in 1000), and the liquor, creosoted or carbolized, is passed through a fine linen mesh and left to deposit. The deposit is collected on a filter of very fine mesh, is successively washed with water to remove every trace of acid, and finally with ether, slightly alcoholized, to remove the fat. The matter when removed from the filter is then dried in a dry vacuum, is agglomerated, and brownish in color.
The moist fibrinous microzymas, completely drained, are composed (in hundredths) as follows:
Organic matter chiefly albuminoid........... 13.553 Mineral matter........................................ 0.384 Water (by difference).............................. 86.063 100.000
Like all organized beings, they contain mineral matter and much fixed water. Their organic matter is chiefly albuminoid; in fact, dry, it dissolves in fuming hydrochloric acid, developing when hot a violet color; and if to the hydrochloric acid water is added a white precipitate of albuminoid matter is obtained.
The minuteness of these humid microzymas, swollen with water, is extreme. Under the microscope they appear to be spherical in form, animated with the brownian movements, the diameter whereof hardly attains 0.0005 (half a thousandth of a millimeter).
Their quantity is very small. From some determinations, unavoidably somewhat uncertain, I estimated that the humid, drained fibrin of the general blood of an ox yields about one thousandth of its weight of microzymas dried at 100°. Taking this figure of 1/1000 as the best approximation, and considering that 1,000 grammes of drained fibrin contain 193 grammes of fibrin dried at 100°, it is evident that the weight of the dried microzymas is 1/193 of the dry fibrin; in short 100 parts of fibrin dried at 100° contain 0.518 parts of microzymas dried at the same temperature. This quantity appears to be very small, and one might think that in the blood it might be neglected, and that consequently the microzymas take no part in the phenomena studied. It is not so, for we shall see that they are anatomical elements and physiological agents of rare energy; and that if it was inter­esting to weigh them, it is still more so to count them.
Let us first show that in the fibrin they are at the same time that which liquifies starch and from which bacteria are derived, that which decomposes oxygenated water, and that which determines its apparent solution in dilute hydrochloric acid.
1. The fibrinous microzymas liquify starch and then become bacteria.
The microzymas of 60 grammes of fibrin obtained from the blood of an ox or of a dog, fresh, still humid, well washed so as to remove every trace of acid, are sufficient to liquify 50 grammes of potato starch at 45° to 50° C. (= 113°-122° F.). The liquifaction is completed in 16 hours; if the reaction is prolonged, Fehling's reagent is reduced. Other things equal, the liquifaction is more rapid with the microzymas of the fibrin of a dog. Finally bacteria appear, while another fermentation begins and the liquid becomes acid.
To estimate the influence of the concentration of the acid in the extraction of the microzymas the fibrin in another operation was treated with hydrochloric acid, 3 to 1,000. The microzymas did not lose any of their activity.
II. The microzymas of fibrin decompose oxygenated water.
The humid microzymas, crude, or with the fat removed by ether, as well as such as have been dried in a dry vacuum, decompose oxygenated water, setting the oxygen free, but with much greater energy than the fibrin from which they had been obtained; showing therein an energy hardly less than bioxide of manganese. I ascertained that the microzymas of the fibrin of the blood of all the animals examined by me acted in like manner.
Later the theory of these facts will be explained, but to anticipate the objection regarding the germs of the air I call attention to the four facts following:
1. Fibrinous microzymas which have liquified starch are still able to decompose oxygenated water;
2. Fibrinous microzymas which have exhausted their decomposing action upon oxygenated water can no longer liquify starch and do not develop into bacteria.
3. Fibrinous microzymas which have been subjected to boiling at 100° C. (= 212° F.) do not liquify starch and do not decompose oxygenated water;
4. Fibrinous microzymas lose the property of decomposing oxygenated water with lapse of time.
But fibrinous microzymas washed in ether, so as to remove their fat, dried in vacuo and protected from contact with the air in a closed tube, preserve for a long time the property of decomposing oxygenated water, but lose by degrees their energy; after ten years they had lost it altogether, without having appreciably lost weight.
Here was another essential property of the microzymas which I formulated!
III. Fibrin owes to its living microzymas the faculty of being dissolved in very dilute hydrochloric acid.
Bouchardat, following Thenard, observed that before dissolving in dilute acid fibrin swelled up in a translucent, colorless gelatinous mass1 and that solution was effected only after prolonged maceration. The progress of solution is so slow that Liebig, for a long time, held that fibrin was insoluble in dilute hydrochloric acid; and we shall see that it was upon this remark that he founded the distinction between muscular fibrin (masculine or syntonine) and blood fibrin. Dumas, on the other hand, verified the fact of solubility and showed that at the temperature of 40° C. (= 104° F.) solution was more rapid. According to Dumas the phenomenon is a function of time and temperature. I shall prove that it is at the same time especially a function of the activity of the microzymas.
1. It was this gelatinous mass thai Thenard correctly regarded as a hydrochlorate of organic matter.
First let us remember that creosote or phenic acid delays the souring and coagulation of milk as well as the vibrionian evolution of its microzymas. Phenol similarly retards the supposed solution of fibrin in very dilute hydrochloric acid. The following will demonstrate the fact:
A mass of 600 grammes of fresh and humid fibrin of ox's blood is divided into four equal parts—A, B, C, D, each of 150 grammes, which are treated in flasks of like capacity in the following manner:} A, 2,000 c.c. of hydrochloric acid, 2 to 1,000; B, the same volume of acid and 40 drops of phenol; C, the same volume of acid and 60 drops of phenol; D, 2,000 c.c. of boiling distilled water. The ebulition is maintained at 100° C. for two minutes. Left to cool and 4 c.c. of fuming hydrochloric acid are added, so that it was also diluted to 2 to 1,000.
The four flasks were covered and placed in the same enclosure; temperature kept at 24° to 28° C. (= 75.2° to 82.4° F.).
In A, B, C the fibrin swelled into a gelatinous mass. In d the fibrin remained a dull white, without becoming gelatinous. In A, the gelatinous mass was dissolved in three days. In B, the solution was effected in four days. In C, the solution was effected in six days. In D, the unswollen fibre remained a dull white; there was no change at the end of a fortnight, though with free access of air.
The phenomenon at the same temperature is then a function of time; it must also be so of the microzymas, since phenic acid retards it the more, the greater the dose, even as it delayed the coagulation of milk, and finally boiling for a sufficient time prevented it entirely, as it had prevented the fibrin and the fibrinous microzymas from liquifying starch and from decomposing oxygenated water. The function ascribed to the microzymas will be made still more clear when it is shown that that which is called the dissolving of fibrin is really the result of a reaction of a profound trans­formation undergone by that part of the fibrin which is in solution. The theory of the phenomenon will also be ex­plained presently; at present we will confine ourselves to saying that in the order of the ideas of these experiments the supposed solution in very dilute hydrochloric acid is, at bottom, only a mode of spontaneous alteration of the fibrin under special conditions. We have now to consider the normal method of its spontaneous alteration.
IV. Fibrin changes spontaneously without undergoing fetid putrefaction.
Gay-Lussac had observed that fresh fibrin, in an open flask, in contact with water which was renewed from time to time, putrefied and disappeared almost wholly, leaving only an insignificant insoluble residue. At the time this observation was made, it was believed that albuminoid proximate principles, as well as others, were spontaneously alterable. This was before the experiments of Schwann regarding the influence of the germs of the air. On the study of this change being again taken up to determine its products, among those which are dissolved, there was observed an albuminoid matter coagulable by heat, which was taken for albumin, also leucine, valeric acid, butyric acid, hydro-sulphate of ammonia, etc. In reality, in the experiment of Gay-Lussac, the alteration was a complex phenomenon, in which the ferments born of the germs of the air take part, and which are the agents of the fetid putrefaction. If the influence of these germs be annulled, the result is different. A mass of fresh fibrin, prepared with the usual care, was immediately im­mersed in distilled water (first carbolized by 3-4 drops per 100 c.c.), so that it was covered with a bed of liquid. Under these conditions, after five to six weeks, at a temperature ranging from 15° to 25° C. (= 59°-77° F.) the fibrin had disappeared; in its place were a clear transparent liquid and a considerable precipitate. No odor except that of the carbolic acid; no vibrios either in the liquor or in the precipitate. The alteration then had taken place without any fetid putrefaction. What was its nature? Later in chapter 11 the nature of these dissolved bodies will be compared with those of the change of fibrin in dilute hydrochloric acid. Let us see of what the precipitate consisted.
The molecular granulations of the change without fetid putrefaction of the fibrin:
In the precipitate, which is greater than the precipitate of microzymas after the disappearance of the fibrin in dilute dydrochloric acid, the microscope shows us a very great number of very small spherical molecular granulations, much more bulky than the fibrinous microzymas, and some shapeless remains, probably of fibrin or of the envelopes of blood globules. To procure these molecular granulations pure the precipitate, which is thick, is steeped in water slightly carbolized, then passed through a close-meshed silk cloth, purified again by levigation, collected on a filter, to be there again washed with water and finally with ether slightly charged with alcohol to remove the fat, and then again with water.
In this condition the molecular granulations preserve their form; they decompose oxygenated water, liquify starch and again decompose oxygenated water after having effected this liquifaction; in short, they possess the properties of fibrin and of its microzymas, but they are neither fibrin nor its microzymas.
In fact, these molecular granulations, the insoluble remains of the disappeared fibrin, treated with hydrochloric acid (2 in 1,000), dissolve much more rapidly than the fibrin, leaving undissolved microzymas identical with, as slender as, and endowed with the same properties as, those of fresh fibrin.
This last observation is important. It is a consequence of the fact that fibrin, under the conditions of the experiment, alters spontaneously without fetid putrefaction, without vibrios, leaving a residuum of molecular granulations which contain microzymas identical with those obtained from fibrin treated with dilute hydrochloric acid. It is explicable only in one way. As milk, treated with a sufficient dose of phenic acid, becomes changed otherwise than milk not so treated or only slightly so, without the microzymas becom­ing vibrios, so the microzymas of the fibrin have trans­formed, in a certain way, the intermicrozymian substance which is in it, as gangue, without undergoing vibrionian evolution, but remaining enveloped as in an atmosphere of albuminoid matter insoluble in water, but easily soluble in very dilute hydrochloric acid, the microzymas being set free.
The great importance of taking these molecular gran­ulations into consideration will be seen when studying in the third chapter the state of the fibrin in the blood. Meanwhile, the fact that the fibrin changes spontaneously in carbolized water, that is to say, without the aid of germs of the air, is a fresh proof that fibrin is not a proximate principle.
In the next chapter we shall see what is the nature of the albuminoid matters of the spontaneous alteration of fibrin in carbolized water, and compare it with that of the change which takes place under the influence of hydrochloric acid.
Meanwhile, such are the proofs, all agreeing, founded on the new method of investigation, to the effect that fibrin, like milk, the liver, etc., is neither a proximate principle nor a compound of such principles, but that like them it is an organic body, containing special microzymas; and further that these living microzymas are what, in the fibrin, liquify starch and can become vibrionien by evolution, decomposes oxygenated water; determines the change of this fibrin either in very dilute hydrochloric acid or in carbolized water.
To complete the history of the microzymas of the fibrin, we must try to discover by what mechanism they decompose oxygenated water and liquify starch, cither isolated or in fibrin; and how it is that they are the agents which determine the spontaneous alteration of fibrin, both in very dilute hydrochloric acid and in carbolized water.
Theory of the decomposition of oxygenated water by fibrin and by the fibrinous microzymas.
I stated at the commencement of this chapter that Thenard, having discovered that organic tissues (for example, the liver) decompose oxygenated water, thought that fibrin decomposed it through being a proximate principle and was the only substance of this order that did so. But what is really the nature of the phenomenon of this decomposition? Thenard said that fibrin and organic tissues "decompose oxygenated water in the same manner as metals (platinum, for instance) without giving up any of their principles, without absorbing the smallest quantity of oxygen, without undergoing the least visible change." In short, that oxygenated water is decomposed by fibrin owing to what has since been called "action through presence," "catalytic action of contact," such as metals or the bioxide of manganese. Such was the state of science a few years ago and is so, perhaps, today. It was necessary for a more exact knowledge of the blood and of organization in general to fix exactly the meaning of this, both as to fact and as to principle; the more so that they were advanced by Thenard himself as a possible explanation of the phenomenon of fermentation, and were the point of departure of the hypothesis called actions of presence, of catalytic contact, which have been the cause that the true theory of fermentation has been so much misunderstood.
In reality, the decomposition of oxygenated water by fibrin, with disengagement of oxygen, is the result not of an action by presence merely, as with the bioxide of manganese, but of a chemical reaction, as is evident from the following experiments:
30 grammes of fibrin of fresh ox-blood, containing 3 gr. 79 of matter dried at 100° C., have successively decomposed three times 60 c.c. of oxygenated water at 10.5 volumes of oxygen. At the second and third addition, the disengagement became gradually slower, so that at the third, after twenty-four hours, no more gas was given off, although the oxygen­ated water was not all decomposed. Altogether 1,600 c.c. of oxygen were set free from 1,890 c.c., which the 180 c.c. of oxygenated water employed, contained. It is evident that if the fibrin had given up nothing, if there had not been some reaction, the successive liquors resulting from the action of the oxygenated water ought not to contain any organic matter. But these liquors on being evaporated left a combustible residue, whose weight,—deducting the ashes,— were 0.16 grammes dried at 100°---i.e., 0.533 for 100 of humid fibrin or 2.76 per cent, of fibrin dried at 100° C.
The fibrinous microzymas also yield up somewhat of their substance in decomposing oxygenated water. Six grammes of these microzymas, fresh, humid, containing 0.84 gr. of matter dried at 100°, having exhausted their decomposing action, the evaporated liquors have left as resi­due, dried at 100°, 0 gr. 06 of organic combustible matter, deducting the ashes; that is to say, 1 for 100 of humid matter-that is, 7.5 per cent, of the weight of the dried microzymas.
Fibrin and its microzymas then do not decompose oxygenated water in the same manner as do platinum or the bioxide of manganese, since they both give up part of their substance which is found transformed in solution in the oxygenated water. If Thenard thought that fibrin gave up nothing it was because, on the one hand, he took into account only the disengaged oxygen, which seemed to him the whole of that which the oxygenated water could furnish; that which had been absorbed being very minute, and, on the other hand, that the fibrin seemed to him not to have undergone any change. But the change really has been great, since that which remains has no longer any action on oxygenated water, does not liquify starch and does not yield bacteria.
These remarks apply to the microzymas which are recovered, similar morphologically, to what they had been before being treated, but do not now liquify starch nor become bacteria by evolution.
It is then a fact, decomposition with oxygen set free from oxygenated water by fibrin or by its isolated microzymas, is correlative with a chemical reaction, with a change in the property of the substance which has exhausted its decomposing activity. And on comparing, in hundredths, the quantities (of the products of the reaction) which are dis­solved, of the fibrin and of the isolated microzymas, it is found that the latter furnish much more than the former. They furnish much more, even if we consider only the quantity of microzymas contained in the fibrin used, viz., o gr. 0335 for 60 grammes of humid fibrin or 5 gr' .79 of that dried at 100° C. In fact, if the dried fibrin yields or contains 2.76, calculating that which its microzymas would give by comparison with what is given by the isolated microzymas, it is found to be 4 per cent, instead of 7 per cent., which is that given by these latter. I do not lay much stress on this differ­ence because it may be due partly to the difficulties and uncertainties attendant upon the weighing. But none the less by means of these comparisons it is clear that the microzymas, whether isolated or not, give up more than does the fibrin, which tends to show that the intermicrozymian gangue of the fibrin does not exert any decomposing action upon oxygenated water, as will be presently directly demon­strated. Anyway it is evident that some substance belonging to the organization of the microzyma—probably a proximate principle—is yielded and transformed, and that it is not the entire microzyma which is the agent of the decompos­ition, since the greatest part of its mass remains, preserving its form. But what is this substance? Without being able to define it exactly we shall see that it is essentially albuminoid. Whatever it may be, it is important to know that it only effects the decomposition on certain conditions. For instance, oxygenated water, which contains a free acid, is not decom­posed either by fibrin or by the fibrinous microzymas, and, reciprocally, the fibrin dissolved by hydrochloric acid under the same conditions as the experiment of Bouchardat, in which the acid is very dilute, and containing the microzymas, decomposes it only when it is neutral. But albuminoid mat­ters combine with several acids; it is without doubt a hydro-chlorate, a sulphate, etc., of this substance, whatever it may be, which is not changed by oxygenated water, and oxygen­ated water which is not decomposed by it. In illustration of this the following very interesting case of the influence of a special acid is given.
Liebig observed that fibrin steeped in a very dilute solution of hydrocyanic acid did not decompose oxygenated water. The observation was true but incomplete, as the influence of this acid is only temporary. In fact, if the quantity of oxygenated water is sufficient, the liberation of oxygen recommences at the end of a time, the longer the greater the quantity of hydrocyanic acid. The decomposition recommences because the oxygenated water destroys the hydrocyanic acid by a phenomenon of oxydation without liberation of oxygen.1
1. C. R.. Vol. XCV, p. 926 (1887). I have since further studied this subject. Hydrocyanic acid and oxygenated water react upon each other; at first without the liberation of gas; an oxamide is formed which crystallizes; at the same time there is a liberation of heat, which increasing, the oxydation is accomplished with production of urea and liberation of oxygen. It is then solely because hydrocyanic acid and oxygenated water react first of all, that the microzymas of fibrin are protected, and not as has been supposed, because hydrocyanic acid acts as a poison upon these microzymas. The fact that after the destruction of the hydrocyanic acid, the fibrin again decomposes oxygenated water, proves that what happens is not a phenomenon of poisoning. This will be further treated hereafter.
Theory of the liquifaction of fecula starch by fibrin and by the fibrinous microzymas.
Fibrin and its microzymas are insoluble in water; on the other hand, Payen demonstrated that fecula exists in a special condition of hydration and of swelling, in which it is similarly insoluble.
How then can these insoluble bodies act upon one another, the one, fecula, dissolving, while the other, the fib­rin or the microzyrnas, remain insoluble? The explanation is the same as before given of the inversion of cane sugar by moulds, born of the germs of the air in its aqueous solution, but which are insoluble, as are the microzymas.
I have proved directly that these moulds, born of other organized ferments, and other microzymas, produce in themselves and secrete soluble products of an albuminoid nature, which are of the same order as those called soluble ferments and which were confused in the same category with organized insoluble ferments. Having thus established the anatomical origin of soluble ferments, to mark the union of dependence between the product and the producer, I gave the name of zymas to what had been termed soluble ferment.
This established, as the microzymasa of sprouted barley produce diastase or hordeozymas, as the pancreas or its microzymas produce pancreazymas, which liquify and saccharify starch, so the fibrinous microzymas produce the zymas which effect its liquifaction.
[aFor the purpose of continuing "the conspiracy of silence" beneath which the marvellous discoveries of Bechamp have been obscured for so many years, this word and its congeners are never used in the writings of the chief conspirators, nor in those of the numerous leaders of the profession who have been duped by them.—Trans.]
And since every zymas is of the albuminoid order, as the fibrinous microzymas which have exhausted their decom­posing action upon oxygenated water do not liquify starch, we can say that the substance which in the fibrinous microzymas is given up and transformed by oxygenated water, is precisely this zymas, an albuminoid substance which liquifies the starch of fecula.
Theory of the spontaneous alteration of fibrin, whether in very dilute hydrochloric acid or in carbolized water.
The two constitutive parts of fibrin are equally insoluble in water and in very dilute dydrochloric acid. As for the liquifaction of starch by them, the same question arises: how can these two insoluble bodies act upon one another, the one remaining insoluble, the microzymas; the other, the albuminoid matter, entering into solution? The answer is the same. In the same manner that fecula is made soluble and transformed by the zymas which the microzymas secrete, so the albuminoid matter is dissolved by this zymas while being transformed.
The explanation of the phenomenon is thus very simple. Only in the case in which very dilute hydrochloric acid inter­venes does the transforming chemical action of the zymas secreted by the microzymas act upon the insoluble combination, which the albuminoid matter makes, at first with the hydrochloric acid; while in the carbolized water it acts directly on the insoluble albuminoid matter as on the amylaceous matter of the starch. But the action of the zymas being exer­cised on the one hand on the hydrochloric combination of the albuminoid matter and on the other on this matter itself, it is not to be wondered at that the soluble products of the reaction differ in some respects, as will be explained in the second chapter.
It is now easy to understand why the previous coction of the fibrin hinders alike its solution in dilute hydrochloric acid and in carbolized water. It is because heat at 100° kills the microzymas as it destroys the activity of all zymases, and doubtless because the inter-microzymian albuminoid matter has undergone the special coagulation which hinders it from effecting the gelatinous combination with hydrochloric acid before spoken of.
To sum up: Fibrin is not a proximate principle. It decomposes oxygenated water correlatively to a change in the zymas produced by its microzymas, which zymas is the agent of the liquifaction of starch and of the changes under­gone by its albuminoid matter, whether in dilute hydrochloric acid or in carbolized water, conditions in which its micro­zymas do not undergo vibrionian evolution. In short, these microzymas, whether in the fibrin or isolated, are not the agents of the decomposition of the oxygenated water after the manner of ferments—that is to say, physiologically by a phenomenon of fermentation, but only as producers of the proximate principle which the oxygenated water changes as it changes hydrocyanic acid.
To complete a knowledge of fibrin and of its micro-zymas, I recall the facts that Estor and I, in our note, described an experiment from which we concluded that "in the presence of pure calcic carbonate and so long as the microzymas of the fibrin continued to evolve they behaved, as regards fibrin, at the same time as alcoholic ferment, and as acetic, lactic and butyric ferments.1 Among these experi­ments I will describe two, because they were conducted on a sufficiently large scale the better to establish results. The proportions of the materials employed were as follows:
Fecula of potatoes, 5 parts, transformed into starch in 85 per cent, of water; pure calcic carbonate, 1 part; fibrin fresh, moist, newly prepared, 0.13 parts; temperature of the oven 35° to 40° C. (= 95°-104° F.).
1. C. R., Vol. LIX.715-716.
The two experiments were started on the 22nd of May. The next day disengagement of gas commenced, a mixture of carbonic acid and of hydrogen. From the 8th day the gas was analyzed repeatedly, and was found to be composed as follows, in hundredths:

gaseous mixture is thus seen to have varied with the complication of the reaction.
One of the experiments was stopped on the 10th September for the purpose of making the analysis. There was still a large amount of fecula not transformed; the products of the fermentation were as follows:
Absolute alcohol............................................................ 21 cent, cubes. Proprionic acid ................................................................... 12 grammes. Butyric acid .............................................................................. 150 " Chrystallized acetate of soda ................................................. 650 " Chrystallized lactate of chalk.................................................. 709 "
The second operation, upon a greater scale, was continued until the lactate formed had been transformed; the analysis of the products was made on the 10th of May of the following year. The experiment then had lasted nearly a year. There was still some fecula not transformed. There were found:
Alcohol mixed with higher alcohols........................................ 78 c.c. Proprionic acid.........................................................................80 gr. Butyric acid ............................................................................680 gr. Acids higher than the butyric up to caprylic ......................... 245 gr. Crystallized acetate of soda....................................................725 gr.

Thus, as in the classical lactic fermentations, the ferment which produced the lactic acid is that also which destroys this acid in the lactate of lime. It is only necessary to observe that the products formed by the microzymas of the fibrin differ greatly, both in proportion and in quality, from those of ordinary lactic fermentations, and especially from those by mother of vinegar. I shall, by and by, insist further on the fact that the bacteria of the microzymas which evolve in the first phase have gradually but completely disappeared in the second, so that at the end there only remained a few forms closely allied to the microzymas.

But I insist here on the fact that for the two experiments 200 grammes of fresh fibrin were employed containing at the start at most 0 gr. 2 of microzymas to effect the prodigious transformations of the fecula. The fibrinous microzymas are then figured ferments of rare energy.

Such were the preliminaries to the discovery of the third anatomical element of the blood. For a complete understanding of the fibrin and the products of its changes it is necessary to know in what light to regard the albuminose of Bouchardat, which this savant believed existed in the supposed solution of fibrin in dilute hydrochloric acid, and to do this we must have a better knowledge of the albuminoids.

CHAPTER V.: The Blood and the Third Anatomical Element by Antoine Bechamp

OF THE REAL NATURE OF THE BLOOD AT THE MOMENT OF A GENERAL BLEEDING. THE LIVING PARTS OF THE BLOOD. PROTOPLASM. THE UNCHANGEABLE CHARACTER OF MIXTURES OF PROXIMATE PRINCIPLES. THE VITELLIN MICROZYMAS AND THE BLOOD GLOBULES. THE VASCULAR SYSTEM. THE BLOOD A FLOWING TISSUE.
The blood really contains three kinds of anatomical elements: the red globules, the white globules and the microzymian molecular granulations. Anatomically, the blood is constituted by three sorts of figured elements and, by a fourth term, a liquid. Is this the serum, this liquid which is its interglobular and intergranular substance?
The three sorts of anatomical elements are living, insomuch as they are organized and contain microzymas which I have proved to be living by their function as ferments and by their capacity to become vibrioniens by individual evolution, which was a novelty for physiology and even for chemists.
Nevertheless, so far as concern the red globule, since 1846 the statement that it is alive was not a novelty. In fact, in a memoir,1 which merits the more attention that it is never quoted, J. B. Dumas made an observation which must be regarded as of the first importance. It is that to isolate the red globules in their integrity, by mixing the blood with sulphate of soda, a current of air must be introduced; without this they will change, losing their coloring matter which itself also changes. And he said: "The globules of blood act as though they were really living beings, capable of resisting the solvent action of sulphate of soda so long as they are alive, but yielding to this action so soon as they have succumbed to the asphyxiation which affects them by the deprivation of air, and which manifests itself with singular rapidity, either by their change of color or by their rapid solution." Dumas asserted clearly that the globules breathe; that account must be taken of their membrane in explaining the phenomenon of respiration; and that the breathing of an animal has especially for its object to furnish oxygen to the globules of its blood and to expel "the products into which they convert it." He also remarked that in the discussions and the calculations respecting respiration the blood had always been regarded as a homogeneous liquid, while it was only the serum which possesses this quality. He in no wise disregarded the part taken by the serum in the phenomenon of arterialization, but he insisted on the preponderant part taken in it by the red globules.
1. Dumas, "Recherches sur le sang," C. R.. Vol. XXII, p. 900 (1846).
To understand the blood, one must place oneself in the order of ideas of the memoir of Dumas, but broadened; that illustrious savant did not recognize in it, nor did any one else at that time, other anatomical elements than the globules, but there is another. He saw in the blood only three nitrogenous organic matters: albumen, fibrin and the globules, but there are others.
I will add that, in the serum, he made allowance for the share therein of the phosphates and other mineral matters.
At the moment of a general venesection the blood has been regarded as being that which it is in the vessels while it circulates in them, but as being a mixture of the arterial and venous bloods; and we have seen that at this moment the blood is so thoroughly regarded as being alive that it was regarded as certain that coagulation was its death.
The blood being alive, it is necessary to recognize, in accordance with the doctrine of Bichat, that, as in all the rest of the organism, the only things living in it are the anatomical elements, that is to say, that of the four parts which constitute it, the three kinds of anatomical elements are the only things living in it; the fourth, the serum, or that which will become the serum, the interglobular and intergranular substance, fulfilling with regard to them only one of the conditions of existence.
But as this conclusion conflicts with the prejudices of the schools, it is necessary to know what those prejudices are to combat them, for they are the negations of the doctrine of Bichat and precisely contrary to it. In fact, while it is asserted that the globules of blood, in general the anatomical elements, are only organites, neither plants, nor animals, as M. Pasteur said, that is to say, not living although organised, it was insisted that that which in the blood is still called plasma was living, a liquid whereof all the materials are said to be in a state of perfect solution, that is to say, without any anatomic, figured structure. But it is well to repeat that such was the state of science just as it was before Lavoisier and before Bichat, when the philosophical naturalist, Charles Bonnet, speaking of the organization, called it "the most excellent modification of matter." Even in France a conception more or less analogous to it, that of protoplasm, was preferred to the striking conception of Bichat. But protoplasm or its synonym, blastema, was considered to be organized living matter without structure. Here is one of the most precise descriptions of such matter: "A completely homogenous, amorphous matter without structure can be regarded as organized substance if it is constituted of numerous proximate principles, united molecule to molecule by special combination and reciprocal solution, and however simple may be this organization, it is sufficient to enable one to say that it is alive." Diet, de Med., Littre et Robin, art. Organique (1878).
Van Tieghem said: "Protoplasm is a mixture with water, of a greater or less number of different proximate principles, in the course of continual transformation."
Huxley said: "All protoplasm is similar to protein—all living matter is more or less similar to albumen."
Cauvet said: "Protoplasm is a nitrogenous liquid, more or less flowing, composed of a translucent joining substance and of fatty and albuminoid granulations."
Even Claude Bernard said: "In its simplest condition life, contrary to the idea of Aristotle, is independent of all special form; it resides in a substance defined by its composition and not by its shape; the protoplasm."
Pasteur said: "Living organisms are composed of natural substances such as life elaborates them, the proximate principles of living bodies which possess faculties of transformation which are destroyed by boiling."1
1. C. R., Vol. LXXIII. p. 302. See letter of M. Pasteur to M. Donne. M. Pasteur's manner of thinking was still that of Chevreul [born in 1786. was still alive and active in 1856]— at the time (1810) of the foundation of his chair at the Museum; Chevreul said, speaking of living bodies, that they are organic bodies in contradistinction to inorganic bodies, which we term minerals. Buffon called minerals gross matter, admitting that there was a universally diffused organic matter which he termed organic molecules, but Buffon wrote before the time of Lavoisier. Chevreul spoke of the proximate principles of organic bodies which are the products of life. Pasteur, speaking of the same proximate principles, says that they are natural substances elaborated by life, which have powers of transformation, etc. It may thus be truly said that there was no idea of life as bound to a determined, structural form of living anatomical elements, according to the conception of Bichat. It is thus to be understood how M. Pasteur could class in the same category, as organites, the red globules of the blood and grains of starch. It is true that the amylaceous granule had been regarded as being a vesicle, but Biot and Payen had shown that it was solid throughout its mass, and I have proved, in my researches upon fecula, that it had neither tegument, nor microzymas, being wholly formed of amylaceous matter contaminated with a trace of albuminoid matter. In the microzymian theory it is not life which produces or elaborates the proximate principles, but the anatomical elements are constituted into living apparatus by the microzymas, according to the same mechanism by which the fibrinous microzymas cause starch to ferment, and elaborate the numerous proximate principles which I have described as produced in that fermentation.
These quotations are sufficient. Protoplasm is regarded as a pure mixture of proximate principles, that is to say, of materials of a purely chemical order. M. Cauvet and others, M. Frey, for instance, have observed the granulations of the protoplasm, but they were supposed to be pure proximate principles. This mixture was declared by some, as in the course of continual transformation; by M. Pasteur, as endowed with faculties of transformation, but without other proof of what is precisely the point in question, viz., whether such a mixture can spontaneously change, can alter itself, give birth to any living being whatever, be it a cellule or a microzyma. If protoplasm were that which it was thought to be, the conception of Bichat would be purely chimerical.
I have incontestably demonstrated, in contradiction to the theory of protoplasm and against M. Pasteur, that every mixture, artificial or natural, of real proximate principles, with water, is, by itself, in every way unalterable, incapable of giving birth to anything living; in short, as not being in the course of continual transformation and as not possessing any faculty of transformation capable of producing in it any spontaneous alteration. And if in such a mixture, boiling destroys the "faculties of transformation" of some zymas, this latter had not been produced spontaneously, it was the product of a living organism. In short, if the mixture contains some proximate principle which can be altered by oxygenation, by absorbing oxygen from the air, this principle is itself the former product of a living organism through the reaction of a zymas.1 I have given positive proof of all of this while studying the conditions of the spontaneous coagulation of milk, which was said to be a pure mixture of proximate principles. Cow's milk, creosoted by a suitable dose to destroy the influence of the germs of the air and completely protected from all contact with the air, first becomes sour and then coagulates. After which, vibrioniens appear in it. If by filtration, by the process which I have indicated in the case of the blood, both the globules and all the milk microzymas of the creosoted milk are absolutely removed, the limpid liquid which results, containing all the proximate principles of the milk, under the same conditions, does not become sour and consequently neither coagulates nor permits the appearance of the vibrioniens. The "faculties of transformation" then resided in the anatomical element of the milk which had been removed by filtration, and not in the rest of its substance, which maybe called the physiological serum of milk.
1. The zymases are never the products of the spontaneous alteration of an albuminoid matter, but are always the products of the physiological function of a living organism and of an anatomical element in the latter. See the article zymas, "Dictionnaire de la langue francaise." Littre (1869).
The physiological serum of the milk, which has the same composition as blastema or of protoplasm, is then naturally unchangeable and consequently not living.
It is the same with the fourth portion of the blood, which we will call the physiological serum of the latter. And precisely as the anatomical elements of milk are the agents of its spontaneous alteration, because they are living, so the anatomical elements of the blood are, on several accounts, the agents of its spontaneous alteration, as will be proved in the following chapter. But first must be determined the physiological role of this serum, in which are realized the conditions of existence of the anatomical elements, globules and granulations of the blood, while it circulates and after it has been shed.
I understand by "conditions of existence" of an anatomical element (following Bichat's conception), that of the preservation of its physical being at the same time with the integrity of its tegument and that of its content, preserved with its composition unchanged, which it can only be by finding in the medium in which it lives all the materials for its nutrition.
Take, for example, the red globules; we know that in blood, steeped in a certain quantity of water, the soluble contents of its globules are diffused by osmose, the teguments remaining whole; on the other hand, we know that in the same blood, steeped in several times its volume of a saturated solution of sulphate of soda, its globules remain entire, both tegument and content. We can even steep the blood in its own serum, without the globules being altered ; without any trace of the colored content being dissolved. And it is the same with the molecular granulations as with the globules; so that if in blood, steeped in the solution of sulphate of soda, a small part of their albuminoid atmosphere is temporarily soluble, as we have seen, it is absolutely insoluble in the serum and each granulation remains there whole and independent, the same as each globule, and this constitutes one of the conditions of the circulation.
But to understand the circulation and the reciprocal influence of the vessels and of the elements of their content, a slight diversion into embryology is indispensable.
In studying the development of the fowl to ascertain the role of the microzymas of the vitellus in the formation of the anatomical elements and of the organs, Estor and shown1 that the container and the content of the vascular system are born and developed simultaneously with the aid of the microzymas and the unorganized materials of the vitellus. We have never seen globules in the body of the embryo before the establishment of the circulation; they are formed on the spot. Thus the anatomical elements of the tissues of the vessels and the anatomical elements of the blood contained therein are born at the same time, by the microzymas of the vitellus as builders, in the unorganized intermicrozymian medium of the vitellus. Hence it results that the serum of the embryonal blood comes into existence concurrently with the globules and the granulations, having the non-organized parts of the vitellus for their source. To sum up, container and content are born at the same time, develop at the same time, and at the same time become what they are destined to be in the future.
1. C. R., Vol. LXXV, p. 962 (1872). We were led to undertake this embryological experiment as the consequence of the following experiment of which Estor was a witness: The mother of vinegar formed a microzymas, united among themselves by a hyaline intermicrozymian substance, is a membrane of mucous consistence with which we have compared the false membrane called fibrin; but it is so much vegetable that it is hardly nitrogenized. But in the "mother of vinegar," under the conditions in which one forces its microzymas to live, these become by individual evolution bacteria, or by association manufacturers of cellules. It is the same with the microzymas of beer-yeast, which, in certain media, act as lactic and butyric ferments, undergoing vibrionian evolution; while in others they reproduce the cellule of yeast and the normal alcoholic fermentation." * The microzymas then can be manufacturers of cellules by grouping themselves together, and being grouped becoming enveloped with a tegument when the conditions of existence of these cellules are united. And it is precisely this which the vitellin microzymas do during embryonic development. This new theory of the origin of the cellule does not weaken the axiom of M. Virchow: omnis cellulae cellula. One cellule may be derived from another cell according to another mode, that is all. Consequently, when M. Pasteur said that the globule of the blood is an organite incapable of reproduction because it could not be cultivated like beer-yeast, he was mistaken, not knowing any other mode of reproduction. *For the developments of the theory of the microzymas, manufacturers of cellules, see the following publications: "Conclusions Concerning the Nature of Mother of Vinegar and of Microzymas in General," C. R., Vol. LXVIII, p. 877 (1869); "Researches on the Nature and Origin of Ferments. Ann. de chemie et de physique," 4th series. Vol. .XXIII. p. 443. And for the theory in its entirely: "Les Microzymas Builders of Cellules." see: "Les Microzymas," etc., M.Chamalet, 60, passage Choiseul, Paris, p 431-463 and p. 948.
The blood ought to be studied not only by itself, but as being to the vessels that which the content of a cellule or of an organ is to its tegument. The tegument of the vascular system consists of the various tissues of the arteries, of the veins and of the capillaries. It must also be borne in mind that the system is directly in relation with the heart, the lungs, the liver, etc., and that the lymphatics (the chyle vessels) communicate directly with it. And as the content of a cellule, of an organ, does not exist without the container, so also the blood does not exist without the vessels which contain it and which make of the whole system an organ in more or less direct relation with every part of the organism.a And it must be observed that if there is any difference between the anatomical constitution of the container of the various regions of the vascular system there is also a difference in their content. Independently of the color there is more oxygen and less carbonic acid in the arterial blood than in the venous. In several regions differences have been observed in the portion of the number of blood globules to that of the leukocytes. Lehmann observed that if the blood obtained from the portal vein gives fibrin by whipping, that of the suprahepatic vein does not furnish any by this means, proving, as we shall see, that the microzymian molecular granulations of the two bloods differ in something, and Denis has already pointed out that the fibrin of the arterial blood is not identical with that of the venous blood, etc.
[a This original conception throws a new light upon the purpose and relations of the circulatory system, which I hope to enlarge upon in a future memoir.—Trans.]
Consequently it is physiologically evident that the anatomical elements, conceived as being personally and in individually living from whatever part of an organism they may be taken exist there only because the conditions of their existence are found naturally realized there. It is not otherwise with the blood; the conditions of existence of its anatomical elements are only realized, in each point of the circuit, while it is contained in the vessel and circulating.
It is ordinarily said that the anatomical elements swim in the lymph, the liquor sanguinis or the plasma; those who, with Milne-Edwards, admitted the existence of finely divided fibrin, said that it too floated in the serum. Anatomically, may we continue so to regard the reciprocal relations of the three anatomical elements and of the fourth portion of the blood? And is it correct to say that at each point of the blood current there are molecular granulations and globules almost in contact with one another? Is it not more correct to say that the fourth part, the serum, is only the intercellular and intergranular substance of these anatomical elements which hinder their immediate contact, a situation analogous to that which is correctly admitted to exist between the anatomical elements of the other tissues? But, if this relation really exists for the blood contained in the vessels, must we not say that the blood not only is not a liquid, but that it is a tissue like that of the content of the spleen, or of the liver, or of the kidney which are more or less flaccid? The softness of the tissue of the content of the vessels is much greater, that is all; we must then say that the blood is a flowing tissue.
The flowing state of the blood tissue is related at the same time to the soft consistence, gelatinous it has been called, and to the elasticity of the globules, whose tegument is incessantly lubricated by the intercellular liquor; to the much softer consistence of the swollen albuminoid atmosphere of the microzymian molecular granulations whose density is nearly equal to that of the serum; to the absolute insolubility of the globules and of the molecular granulations in the intercellular liquor, which again contributes to their individual independence. This general insolubility of the anatomical elements is assured, at every point of the circuit, by the stability and even the origin of the composition of the very complex intercellular liquor, resulting from the nutritive functioning of the anatomical elements of the container and of the content, and at the same time by the matters contributed by the divers organs with which the circulatory system is in relation, and especially with the respiratory apparatus.
At the moment that the blood is shed it may be regarded as being the same flowing tissue that it was in the vessels. Nevertheless, there is already a profound difference, viz., it is not only a mixture of venous and arterial blood, but of the bloods of all the regions, whose anatomical elements are violently placed in new conditions of existence, very different from their physiological conditions.
We shall see how this change in the conditions of exist­ence rapidly determines the manifestation of the phenom­ena of coagulation and then of other alterations of the blood.

CHAPTER VI.: The Blood and the Third Anatomical Element by Antoine Bechamp

OF THE REAL CHEMICAL, ANATOMICAL AND PHYSIOLOGICAL MEANING OF THE COAGULATION OF THE SHED BLOOD; COAGULATION OF THE BLOOD; THE BLOOD OF THE HORSE; THE SERUM OF THE BLOOD; COAGULATION OF BLOOD DILUTED WITH WATER; SECOND PHASE OF THE SPONTANEOUS ALTERATION OF THE BLOOD; THE BLOOD IN CALCINED AIR; EXPERIMENT PROVING OXYGEN HAS NO SHARE IN THE DESTRUCTION OF THE GLOBULES IN THE DEFIBRINATED BLOOD; SPONTANEOUS ALTERATION OF FLESH; SPONTANEOUS ALTERATION OF MILK; COAGULATION OF MILK; FERMENTATION OF THE EGG; SPONTANEOUS DESTRUCTION OF THE CELLULE OF YEAST; SPONTANEOUS DESTRUCTION OF TISSUES; SPONTANEOUS ALTERATION OF THE BLOOD.

The blood is a flowing tissue; Bordeu had already re­marked that it was flowing flesh. This chemically, histologically and physiologically is far from being true;1 the only thing certain is that the blood like the flesh is a tissue, and that both of them are spontaneously alterable, as are all tissues, when the natural conditions of existence of their anatomical elements are no longer realized. For instance, in the case of the muscular tissue, cadaveric rigidity follows death very quickly, and, in the case of the blood, the formation of the clot follows closely upon its issue from the vessels.
1. It was, I think, in 1742, in his thesis, entitled "Chylificationis historia," maintained, at Montpellier, at the age of 20 years, that Bordeu, among the original ideas which makes him to be regarded as one of the precursors of Bichat, put forth the idea that the blood is flowing flesh. In the seventeenth century, Amyot had already said that "the blood is engendered by the transmutaiton of some flesh which becomes a flowing liquid." (Diet, of Littre.) If an original sketch, later recognised as correct, is sufficient for the author to be historically regarded as the discoverer, assuredly Bordeu would deserve to be regarded as having discovered that the blood, like muscle flesh, is a tissue. But, as observed by Babinet, "if the ancients have said everything, they have demonstrated nothing." Bichat also inserted the blood tissue among his twenty-one elementary tissues, next to his muscular tissues. But since Bichat other savants have so done. In my time at Montpellier the Professor of Physiology, M. Rouget, taught that the blood, because of its globules, is a tissue; and I replied that, according to the ideas then accepted, the blood is no more a liquid or a tissue than was sweetened water holding in suspension globules of yeast. To-day, M. Ranvier also says that the blood is a tissue because it contains figured elements like the lymph. Doubtless the chief condition necessary for its being regarded as a tissue is for a product of an organism to contain some figured element, but that is not enough; according to the doctrine of Bichat it is also necessary to show that this element is living; and still that is insufficient; otherwise milk, the saliva, and even the urine and certain pathological serosities, spontaneously coagulable, would like the lymph and the blood be tissues. I will consider this further in the last chapter.
It is not disputed that the phenomenon of the coagulation of the blood is spontaneous; the standard facts concerning this phenomenon are as follows: The defibrinated blood obtained by whipping does not coagulate spontaneously, and the globules remain intact in the liquor which has lost its peculiar viscosity.
The blood of oxen and of sheep (I leave for the present the blood of the horse), received into a glass or metal vessel, seems to coagulate throughout its mass, uniformly from the periphery to the center, forming a single solid clot which follows the shape of the vessel into which it has been received. This clot contracts by degrees, up to a certain limit, expelling from it in so doing the serum of a lemon color, which thereafter becomes red-colored, getting gradually deeper, so that the contracted clot (withdrawn from the edges) floats in the serum which has been expelled from its primitive mass. As Haller has already said, the clot is formed by the network of fibres of the fibrin which imprisons the globules in its meshes.
It remains to explain these phenomena invoking only the chemical, physiological and anatomical facts studied in the preceding chapters. The necessary condition for the tissue to remain flowing is that the properties of the anatom­ical elements and their independence remain unchanged; that their relations with the intercellular liquor remain con­stant, not only in the vessel, but also after the venesection.
We know the distribution of the globules in the blood, and how they pass, one by one, into certain capillaries; the distribution of the microzymian molecular granulations is such that if the globules should disappear they will occupy all the space which the globules occupied; that is to say, that the former exist in such a manner in the blood that the globule; move in it in displacing the former unceasingly, but in immediately reoccupying the abandoned space; in short they realize the conception of Dumas, when he said of the fibrin that it exists in a flowing condition in the blood; only that this flowing condition is molecular, attached, as we have seen, as to each molecular granulation to a microzyma for nucleus, forming a limited atmosphere around each, which albuminoid atmosphere is absolutely insoluble in the blood serum.
To understand that the number of microzymas of the blood is sufficiently large in order that, surrounded by the atmosphere which constitutes them microzymian molecular granulations, they may occupy every point of the blood mass, even that of the globules which were driven away, it is sufficient to know that they exist there in innumerable quantity. This is proved in the following manner: The fibrinous microzymas, that is to say, the blood microzymas, are with the pancreatic microzymas, the smallest I have observed. They assume, in their extreme minuteness, the spherical form. The diameter of these microzymas probably does not attain 0.0005mm (mm?) in the humid state. This enables us to calculate that in the volume of 1 mm. cube there are at least 15 milliards 250 millions. Now a litre of sheep's blood furnishes 5.25 grammes of dried molecular granulations, which nearly represent the weight of the fibrin that the same-blood furnishes by whipping. But the fibrin, supposed dried, contains 1/193 of its weight of dried microzymas; then 5.25 grammes of molecular granulations, likewise dried, contains 5.25/193 = 0.0272 grammes; that is to say, 27 milligrammes of dried microzymas per litre of blood, which represents a very much greater weight of humid physio­logical microzymas; but in taking this figure for the weight of the microzymas in the physiological condition of humidity, and 15 milliards per milligrammes or cubic millemetre, it is seen that one litre of blood contains more than 27 times 15 milliards of microzymas. But their weight is in reality much less than this, for, humid, these microzymas can retain 80% of water; in the blood, enveloped with an albuminoid atmosphere saturated with the intercellular liquor, they certainly retain less, but in a manner to render legitimate the approximate calculations above given.
It will be interesting to learn the thickness of the albuminoid atmosphere which surrounds each microzyma to constitute the microzymian molecular granulation, such as it exists in the blood at the moment of venesection. An approximate idea of this can be obtained by considering that the volume of the spherical molecular granulations with a condensed atmosphere of the deposit formed in the blood which has had added to it twice its volume of alcohol at 35-40 degrees is about 50 cubic centimetres per 1000 c.c. of blood; making allowance for the space occupied by the globules, we may consider that the volume of the molecular granulations, before the condensation of their atmosphere, was about twenty times greater to occupy the entire space of the 1,000 c.c. of blood; it will be presently proven that they do in reality occupy it all. The albuminoid atmosphere being thus swollen and saturated with the intercellular liquor, it can be understood that the great number of milliards of these molecular granulations arc sufficient to occupy the entire space presented by the blood, provided that their density be very little greater, if not equal, to that of the intercellular liquor which isolates them from one another. This state of the microzymian molecular granulations explains the sort of viscosity which belongs to the blood, and how the globules, whose density is greater, move about in it without being deposited and are only deposited very slowly in the ox's or sheep's blood when at rest; and we shall see how the exception presented by the blood of the horse confirms these considerations.
We have now to inquire whether, after the shedding the blood, the conditions, which I have mentioned as necessary for the blood tissue to remain flowing, can still be realized.
And first it is evident that this tissue, bearing in mind that we are considering a mixture outside of the vessels, longer in its natural physiological situation.
In this new situation the intercellular liquor, in which are united all the soluble organic and mineral products of the denutrition of the anatomical elements of the containers and of the contents, immediately changes its composition; for the disassimilated products, which have become non-usable, are no longer eliminated, and the usable can no longer be utilized or renewed; further, the anatomical elements of the flowing tissue, which have imperative need of oxygen to function properly, are more and more deprived of it; for, after having consumed all that was held in reset the flowing tissue and which the uneliminated products, thus accumulating in it, had been able to absorb, the oxygen thus consumed is not renewed by respiration. The first change then which happens in the shed-blood is that which the intercellular liquor necessarily undergoes in its composition.
The microzymian molecular granulations are first anatomical elements to be affected by this change of medium and of conditions of existence, and, we have seen, this impression is so intense and at the same time so rapid that it manifests itself in a few seconds by the profound change which occurs in the albuminoid substance of their atmosphere which, from being as it was immediately soluble in very dilute hydrochloric acid, becomes insoluble in it, dissolving in it, only as a function of time and temperature, while being transformed. It follows that this influence has the effect of coagulating this substance relatively will dilute hydrochloric acid.
That settled, the mechanism of the formation of the clot is as follows:
The microzymian molecular granulations exist throughout the whole of the space occupied by the flowing tissue, excepting that which is occupied by the globules and the intercellular and intergranular liquor. Thanks to their density, though very little greater than that of the intergranular liquor, they approach one another and come into contact when at rest; their albuminoid atmospheres, soft and mucous, mingle together, while at the same time their substance undergoes the coagulation of which I spoke. And these changes are so rapid that the globules, although much superior in density, have not the time to be precipitated and are caught in the meshes of the network formed by the sol­dering of the albuminoid atmosphere which constitutes the fibrin and membranes, as already said by Haller.
Both the molecular granulations and the globules are so closely connected by capillarity to the intercellular liquor that at the time when, or some minutes afterwards, the clot is completely formed, or, as it is said, the coagulation is complete, the vessel containing it can be turned upside down without any trace of the liquid escaping. This, in fact, is what is to be expected from what I have said about the distribution of the molecular granulations in the flowing tissue and of that of the intercellular liquor around the three anatomical elements.
It is true that it might be maintained, with some appear­ance of reason, "but that is precisely what happens in the plasmatic hypothesis." But that hypothesis has never been verified; on the contrary, I have directly proved that plasma does not exist in the blood, but that the existence of molecular granulations with their central microzymas was certain, as was also that of the microzymas in the fibrin obtained by whipping. But the following are two phenomena which the hypothesis of the plasma cannot explain.
The coagulation being complete, by degrees the clot divides itself spontaneously into two parts. That which, in the clot, encloses the globules; that is to say, the network of fibrin formed by the soldering of the microzymian molecular granulations, contracts then more and more, up to a certain limit, preserving the shape of the vessel in which the clot is moulded, and while the retraction takes place a part of the intercellular liquor is expelled, constituting what is called the serum, in which the retracted tissue is now immersed.
And the first serum thus expelled is transparent and lemon colored, but by degrees the oxygen which the intercellular liquor holds dissolved, as well as that which the globules contain, is consumed; then is manifested the phenomenon observed and explained by J. B. Dumas in globules deprived of oxygen; they change, and their changed coloring matter is diffused in the circumambient serum which becomes more and more of a deep red color. This is what the plasmatic hypothesis cannot explain, if one regards the plasma as a liquid in which all the components are in perfect solution. Let us now give a more direct demonstra­tion of this fact.
All other things being equal, the rapidity of the coagulation of the blood may vary notably from one species to another. The blood of the horse, under the same conditions as that of the ox and of the sheep, is well fitted to verify the truth of the role ascribed to the third anatomical element of the flowing tissue. It is known that the (shed) blood of the horse is divided by rest into two layers: the lower, called cruor, is formed by the globules; the upper, called liquor, contains the microzymian molecular granulations. The upper layer, transparent but not limpid, flowing, possessing the peculiar viscosity, can even be decanted after the globules are precip­itated and very quickly forms a clot in all its mass, in such wise, that the containing vessel maybe turned up-side down without one drop of the liquor escaping; after which the retraction, with loss of transparency, is produced and the serum commences to be progressively expressed from it, as in the case of the bloods whose globules do not separate. Now this retraction would not be produced if dealing with a substance really dissolved which, in coagulating, should become insoluble in the same medium.1
1. The coagulation of the blood has been compared to the gelatinization of a solution of gelatine (Frey. Traite d'histologie et d'histochimie. p. 141). but a solution of pure gelatine heated in distilled water and sufficiently concentrated can be obtained absolutely limpid by careful filtration. On cooling this solution forms a jelly, more or less consistent, perfectly limpid, not undergoing any other contraction than that produced by the lowering of the temperature. Nevertheless, in fact the gelatine has coagulated, for in the gelatinized solution it has become insoluble in cold water as it was before. But the comparison made by Dumas with the state of fecula in starch is more correct. In fact in the transparent starch the fecula is not dissolved, it cannot be filtered. By cooling, after a long time, the starch undergoes a change in its appearance; it becomes more opaque and a retraction accompanied by expulsion of liquid can be observed. This happens because the fecula was not dissolved, but simply enormously distended.
The peculiarity presented by the blood of the horse may be due to the greater difference between the density of its globules and that of the intercellular liquor, and at the same time to a greater softness of the albuminoid atmosphere of the microzymian molecular granulations, which will be more swollen, and, consequently, their mass surrounded by the serum more readily traversed by the globules. I have, therefore, compared, as being the only thing accessible for experimentation, the serums of blood of the ox and of the sheep, with that of the blood of the horse, with regard to their general composition.

The most striking thing, above all, is not alone that the serum of the blood of the horse contains less fixed materials, organic and mineral, with a rotatory power very nearly equal to that of the blood of the ox and much lower than that of sheep's blood, but especially that it contains between seven and eight times less mineral matters than either of the other two.
The serum of the blood of the horse differs then prodigiously from the two serums to which I had compared it; this amply explains at once the softness of the albuminoid atmosphere of the granulations and the rapid deposit of the globules.
Experiment thus verifies the fact that the molecular granulations occupy in the blood not only all the space occupied by the globules, but also all the space left free by their precipitation.
According to Charles Robin1 agreeing with many authors, the blood of man, and that of the dog and of the ox, behave like that of the horse, when they are cooled down a little below freezing; they remain liquid a sufficiently long time to enable the globules to be deposited, the leukocytes, according to Donne, forming a grayish layer upon the top of the blood globules; the supernatant liquid then forms the clot, losing its transparency when the temperature reaches 12° to 14° C.(=53°.6 to57°.2,F.), and it should be added that Ch. Robin, having observed the transparency of the supernatant layer separate from the globules under these circumstances, and which he called plasma, stated that it could not be filtered.
1. Ch. Robin, Lecons surles Humeurs. p. 59 (1871).
I regret that I had not had the time to verify these experiments, but the facts may well be regarded as true, being certified by Ch. Robin. They support the theory which I lay down; the lowering of the temperature below zero (32° F.) having the effect of singularly retarding the functions of nutrition of the anatomical elements (as it retarded those of beeryeast) ought to retard the coagulation of the albuminoid atmosphere of the microzymian molecular granulations.
The formation of the classical fibrin by whipping remains to be explained, and this is now very easy. It is the result of a simultaneous mechanical and chemical action. By the mechanical action the layer of intercellular liquor which separates the molecular granulations is broken, while the granulations forcibly set free are united by their mucous albuminoid atmosphere, at the same time that the changes in the conditions of existence determine the allotropic transformation of the albuminoid substance which, coagulated as we have seen, contracts at the same time, still enveloping the microzymas; in consequence of which that which was diffused throughout all the volume of the blood is reduced to relatively small volume occupied by the classical fibrin. And by the small volume of the fibrin obtained by whipping a judgment may be formed of the enormous volume formed by the albuminoid atmosphere which surrounded the granulations of the flowing tissue at the moment of the venesec­tion, as we have observed also in relation to the molecular granulations when separated from the alcoholized blood.
In the separation of fibrin by whipping the globules remain entire, and I have explained that if the weight of the fibrin thus produced is less than that of fibrin obtained by washing the contracted clot, it is because to the free molecular granulations of the blood are added those obtained by the destruction of the globules with their envelopes.
Such were the facts upon which rested the experimental physiological theory of the spontaneous coagulation of the blood, when, in 1895, I communicated them to the Bordeaux Congress of the French Association for the advancement of science.
If there remained any doubt as to the value of the theory concerning the spontaneous coagulation of the blood as a flowing tissue, here is what must remove them. The new experiments to which they relate are the fruit of the following considerations:
If the formation of the clot is actually the result of the spontaneous soldering of the mucous albuminoid atmospheres of the microzymian molecular granulations; if in the presence of alcohol, diluted to a suitable degree, these atmospheres are condensed, the molecular granulations remaining independent of one another, what would happen if instead of alcohol the blood should be received into water? The following is the experimental answer:
Coagulation of blood diffused in water. At the moment of venesection the blood is received into increasing volumes of distilled water up to half of its volume. The clot is formed in all cases; with small quantities the globules do not appear to be altered and the first serum has its ordinary appearance, but in proportion as the quantity of water is increased there arrives a time when the serum becomes colored. Encouraged by these trials, one day in November, at Paris, the blood of the general venesection of a Russian sheep was received, as to one part into two volumes of alcohol at 36°, carbolated with two drops per hundred c.c., and as to the other part into two volumes of distilled water carbolized in the same proportion.
The alcoholized mixture yielded the ordinary deposit of microzymian molecular granulations with the properties with which we are now familiar and naturally there was no trace of a clot.
The aqueous mixture furnished something very different from a precipitate. Like the alcoholic mixture, the aqueous mixture had been made at 9 o'clock in the morning; naturally, like the other, it was of a deep red color, since under such conditions all the haemoglobin of the blood globules had become dissolved. But at 3 p.m. the aqueous mixture was coagulated, the clot occupying the entire volume of the mixture, a volume three times greater than the blood clot without the addition of water. A little deep red liquid was already expelled; the next day the blood clot was not more contracted.
The experiment repeated with ox blood gave the same results; the blood clot was formed in the entire mass, floating in a little deep red liquid, etc.
The trembling and nearly transparent clots of the two experiments were placed to drain on a moist cloth of close texture. At a given moment the cloths were found covered with a mucous substance which, a prolonged washing, with water slightly carbolized (1 drop per 100 c.c.), then with alcohol at 25%, and again with water, did not completely decolor; at the end there remained a red false membrane which could be removed in a single piece from the damp cloth; such was the appearance and state of the fibrin of the clot formed under these conditions; its quantity is percept­ibly that which is isolated by the washing of the ordinary clot. In this condition this fibrin is not dissolved immediately in hydrochloric acid at 2/1000, but like ordinary fibrin is a function of time and of temperature, and without first taking on the condition of a jelly like the fibrin from whipping, etc.
If then the albuminoid atmosphere of the hematic microzymian molecular granulations is condensed under the influence of alcohol of proper strength, the same atmos­phere is much more distended, three times more, in water, and the blood still coagulates in all its mass, the globules being destroyed.
The conclusion which results from this new series of experiments is that the physiological theory of the spontaneous coagulation of the blood, founded on the existence of a third anatomical element, the microzymian molecular granulations of the flowing tissue, sufficiently explains the facts.
It is time then to erase from the language of science the words plasma, plasmine, fibrinogen and of fibrinoplastic, with which it has been encumbered.a There must also be erased for the explanation of the phenomenon the pretended influence of the globules, of calcareous or other salts, catalytic actions of contact, etc., not to speak of various occult
[a. Term microbe must also be erased; unless it be desired to retain it to denote mankind and other short-lived animals! Trans.]
influences. An exact knowledge of the anatomy of the blood and of the conditions of existence of the anatomical elements will suffice.1 But it is also necessary to understand in another way than has heretofore been done the meaning of what is called the coagulation of the blood. In truth, the blood does not coagulate. The experiment proves it; it is the substance of the atmosphere of the third anatomical element of the flowing tissue which, in undergoing the allotropic change of coagulation, gives to the aggregate of the phenomenon the appearance of a total coagulation; but, as we have seen, it is only an illusion.
1. It is because the conditions of existence of the anatomical elements are longer realized when the blood is preserved between two ligatures in the vessel which contains it that the coagulation of the flowing tissue is often long defer red. This explains the success of certain experiments of authors and, the most recent, those of M. Fr. Glenard (1875). (Thesis above quoted.)
The supposed spontaneous coagulation of the blood is at bottom, only the end of the first phase of the spontaneous alteration of the flowing tissue, as the cadaveric rigidity, marks the first phase of the spontaneous alteration of the muscular tissue.
But, what is it that changes in a tissue? And what is the second phase of the spontaneous alteration of the blood and at what moment does it begin?
It is of importance to the plan of this chapter to give a precise answer to these questions.
Second phase of the spontaneous alteration of the blood.
The first begins by the chemical alteration of coagulation of the albuminoid atmosphere of the microzymian molecular granulation, whence results the formation of the clot, the retraction of the latter and the expulsion of the lemon-colored serum. The globules have nothing to do with this phenomenon, as is incontestably proven by the experiment with the blood of the horse.
The second phase begins at the moment when the serum becomes colored with red, which shows that the change of the blood globules has commenced, their haemaglobin, more or less changed, being diffused in the serum. The following experiment of M. Pasteur has shown what becomes of the globules in this change. This savant made it in 1863,1 five years after my verification of the hypothesis of germs in the air, when he had given up belief in the spontaneous generation of ferments, with the object of demonstrating that in the absence of germs the blood would not putrify because nothing living would appear in it. To understand this it is to be remembered that M. Pasteur was a protoplasmist seeing in an organism only proximate principles, admitting in it nothing figured, autonomically living, comparable to the figured ferments.
I take the recital of the experiment from a book of the author, published long after the microzymas had been discovered and the microzymian theory of the organization completed.
He commences as follows: "Let us examine into the interior of living beings, in good health, such or such of the materials which may be found there, and examine them in the state in which life has formed them, in contact with pure air."2
1. Pasteur. "Recherches sur la putrefaction." C. R.. Vol. LV1 (1863). 2. L Pasteur. "Etudes sur la beere." p. 46 (1876).
In fact, with the assistance of Cl. Barnard, he poured the blood of a dog directly into a vessel, the air in which had been calcined. The receiver, sealed with the blow-pipe, contained thus one of the materials to be examined, obtained from the interior of the animal, and thus protected from the germs of the air. I now quote textually as follows what M. Fasten thought he observed:
1. The blood does not putrify, even at the highest temperature of the atmosphere; its odor remains that of fresh blood, or takes on that of lye.
2. After an exposure of the flasks to 25°-30° C. (= 77° to 86° F.), during several weeks, nothing can be observed but an absorption of 2% to 3% of oxygen, which is replaced by a perceptibly equal volume of carbonic acid.
3. Under the circumstances in which the blood of the dog, exposed to the contact of pure air, does not putrify at all, blood crystals are formed with remarkable readiness.
4. In the first days of its being placed in the oven, slowly, at the ordinary temperature, the serum became gradully colored a deep brown.
5. In proportion as this effect is produced the blood globules disappear and the serum and the clot become filled with crystals of a brown or red color. After some weeks not a single blood globule remains either in the serum or in the clot. After a longer interval the whole of the fibrin may become collected in a single hyaline mass.1
1. Ibid, p.49.
Such was the experiment from which M. Pasteur concluded that, if protected from germs of the air, blood did not putrify at all; that is to say, was not altered by the action of any figured ferment, which in his opinion only the germs of the air could produce. I have elsewhere shown that, supposing the technique of the experiment to have been accurately carried out, the observations made thereon were incomplete, and the interpretation of the results vicious in the extreme. I will revert to this hereafter; for the present I will only show that the facts of the experiment corroborate my own.
It is evident that, taken altogether, the experiment of M. Pasteur has confirmed that which the microzymian theory never fails to prove, namely, that every tissue, every humor, withdrawn from a living healthy animal, absolutely protected from the germs of the air, necessarily alters and, consequently, alters spontaneously.
It demonstrates further that there are two distinct phases in the changes of the blood.
It is true that M. Pasteur did not stop an instant to consider the phenomenon of coagulation, but he observed that the serum, at first lemon-colored, became by degrees red, then deep brown, without insisting on the mechanism of the contraction of the clot and the expulsion and tardy coloration of the serum which marks the commencement of the second phase and which follows so exactly the consumption of oxygen which the blood contained, that the author himself testified to the absorption of a small quantity of the oxygen in his flasks with a corresponding production of carbonic acid. During the second phase, in which the haemoglobin alters more and more, blood crystals are formed, at last the glob­ules are destroyed and disappear while the fibrin which imprisoned them in the network formed by it contracted more and more.
This picture shows clearly that during the two phases the alteration is at once chemical and anatomical, ending in the destruction and total disappearance of the globules.
But to what cause did M. Pasteur ascribe such prodigious effects? In 1863 he also experimented on muscle flesh, imitating my method of investigation, replacing creosote by alcohol. He wrapped up a voluminous mass of flesh in a linen, soaked in alcohol, and left it to itself.
"There will not be any putrefaction, he said, not in the interior, because the germs of the vibrios are absent, nor externally because the vapors of alcohol hinder the development of the germs on the surface." Nevertheless the author certified that the meat "became gamey in a pronounced manner." And why did it become gamey? Simply, said he, "because it is impossible at ordinary temperature to withdraw the interior of this flesh from the reaction of solids and liquids upon one another . . . There will necessarily always be what are called actions of contact, which develop in the flesh small quantities of new substances, which add to the savor of the meat their own savor."1
Then at ordinary temperatures the same thing should happen with the blood as with the muscle tissues; there will be actions of contact, reactions of the solids upon the liquids; and that is why the blood changes without putrifying as flesh becomes gamey without putrefaction. There is some excuse for M. Pasteur and the academies with him to have been satisfied with these explanations, seeing that protoplasmism had been accepted as a dogma among savants. It was faith in this doctrine which caused the globule to be regarded as an organite, and the mass of the blood or of the flesh, for a collection of proximate principles, and, which is more ser­ious, hindered M. Pasteur from seeing the microzymas among the results of his experiment, or, if he saw them, caused him to neglect them, as he had neglected the micro­zymas and even the vibrios in the gamey meat. However it maybe, it was by invoking actions of contact, and of reactions of solids and liquids, that M. Pasteur persuaded himself that the alterations of the blood and those of the flesh were not phenomena of putrefaction; that is to say, of fermentation. And this manner of regarding things on the part of the cele­brated savant prevailed so generally that I was obliged to make an explanation on the subject of an assertion made by Balardin 1874.
M. Servel, Estor's assistant, had presented to the Academy2 a work verifying the fact that with absolute protection from atmospheric germs the most divers tissues could
1. L. Pasteur, Recherches sur la putrefaction. C. R., Vol. LVI. p. 118-9 (1894). 2. C. R., Vol. LXXIX, p. 1270.
produce bacteria even in their interior, and cited other verifications made in Germany. Now, Balard, who presented the work, took the occasion to say that blood is preserved without putrid fermentation and without bacteria in the experiment of M. Pasteur.1 I replied to M. Balard, saying that the blood is one of these substances, of which the egg is another, whereof the microzymas undergo with most difficulty the vibrionian evolution.2 In spite of this the experiment of M. Pasteur was given as a pre-emptory demonstration that the interior medium contains nothing figured which could become bacterium by evolution in the substance of a tissue or of a humor; and it was asserted, in agreement with him, that the experiment proved at the same time that the bodies of animals were closed to germs from without.
1. Ibid., p. 12722. Ibid.. Vol. LXXX. p. 494. The note of M. Servel and my reply to M. Balard should be read with attention to have a clear idea of the state of the question in 1875.
Nevertheless, in a discussion at the Academy of Medicine, where, once more, I defended the microzymian theory, M. Pasteur took part in the discussion, maintaining his former conclusions, continuing even to deny the exist­ence of the microzymas. It was then that I urged against him his own experiment upon the blood which is a demonstration against his own system. I said: "Do you affirm that the blood in which crystals are formed and the globules disappear is not altered? The globules of this blood are always destroyed and disappear: what then has destroyed them? Even the haemoglobin is transformed into crystals and we find in the liquid a swarm of microzymas . . . these microzymas which you have neither seen nor noticed."
No longer invoking actions of contact, M. Pasteur said: "But these transformations are made under the influence of the oxygen of the air." With regard to the presence of the microzymas he admitted it in leaving it to be believed that I had stated that they became bacteria in his experiment.1
1.Bulletin de l'Academic de medecine, 2d series, Vol. XV. p. 679.
The presence then of the microzymas being acknowledged, the observation of the results of the experiment was completed, it matters little after that, that M. Pasteur continued to treat them as "creatures of the imagination," and that he explained the phenomena as the result of some influence of oxygen; I had reason to hope that this avowal would open the eyes of my academical opponents and that they would acknowledge that they had been deceived. But nothing of the kind; the acknowledgment exists nevertheless, and it only remains to prove that they are really the agents of the changes of the second phase of the phenomenon and of the destruction of the globules.
In the first chapter we have shown that fibrin left to itself covered by carbolized water, open even to contact with the air, is transformed into soluble products without the appearance of bacteria, leaving a residue of new microzymian molecular granulations without phenomena of fetid putrefaction; and we have also seen that the microzymas of these granulations were the ferments of the transform­ations. On the other hand we have also seen, that in fecula starch, the same fibrin liquefies this starch and makes it ferment, while its microzymas become bacteria; we have there two examples, in one of which the microzymas are active without evolution, in the other they undergo bacterian evolution.
The following experiment demonstrates that oxygen has no influence in the phenomenon of the destruction of the globules in defibrinated blood.
About 300 c.c. of the blood of the ox, having added to it 50 c.c. of a saturated aqueous solution of phenol, were immediately defibrinated and the blood carefully separated from the fibrin submitted to a current of carbonic acid, for the purpose of expelling the oxygen. The flask was closed and left to itself at the temperature of the month of June in Montpellier during one month, and afterwards in the oven at 30° to 33° C. (= 86° to 91°.4 F.); this blood did not undergo fetid putrefaction, the globules, very slightly altered in shape, remained whole during the first ten or twelve days. It was only about the 15th of June that there appeared a great quantity of very fine molecular granulations, of which only some rare examples had before appeared, without any trace of vibrios or of bacteria. The globules resisted for a long time further, and ended by disappearing. Here was an alteration without the presence of oxygen, whereof the microzymas could not be other than those of the globules.
The fibrin and the globules of defibrinated blood can then be destroyed by their own microzymas alone, without fetid putrefaction and without bacteria.
If in the experiment with the blood of the ox, defibrinated or not, no blood crystals were formed it was because the haemoglobin of this blood is one of those which either do not yield them at all or do so with difficulty.1
1 As to the assertion of M. Pasteur relative to the influence of the oxygen of the air, he knew that a long lime before I had refuted this in advance (see Les Microzymas, p. 253. and following (1883),Chamalet, 60, Passage de Choiseul): it is there shown that the blood taken from the crural artery of a dog, with the addition of a little of the saturated solution of creosote, submitted to a continuous current of common air is preserved arterialized and the globules remain perfect a long lime; these last, however, end by being destroyed so that the microzymas become free, often without the appearance of bacteria, and always without felid putrefaction. When the current of air is replaced by pure oxygen the same thing happens and the crystals of blood are formed at between 24° to 26° C. of temperature. It is on the contrary, in carbonic acid, that the blood globules of the dog are destroyed most quickly and the crystals are formed most readily between 35 to 40 C, always without fetid putrefaction. Later I showed that under :he same conditions as that of the blood of the dog, the bloods of the ox. of the pig, of the fowl, of the duck give neither crystals nor yet the soluble haemoglobin of the ox. Bulletin of the Academy of Medicine, 2d Series. Vol. XVII. p. 225 (1887). I will add that under the prolonged action of the current of air on the blood of the dog I have found that the quantity of normal urea was increased. This statement should be verified.
Assuredly, if in spite of the changes certified to by himself, M. Pasteur came to the conclusion that muscle, flesh and blood were not liable to become putrid, it must have been because he firmly believed that ferments had for their only source the germs of the air and that the protoplasmic system of organization was founded on rigorous observation. And I venture to say that he knew he was in error and that, later, it was with design that he disputed the microzymian theory, being unwilling to confess that he had observed badly and had taken the wrong road.
Prof. Joseph Bechamp reviewed the experiment on flesh as I had done for that of the blood. He repeated the experiment of M. Pasteur without using alcohol as an antiseptic, and in the centre of the piece of meat, there where M. Pasteur said that the germs of vibrios were absent, he found the microzymas in evolution and vibrios or bacteria. At the same time he found the tissue disorganizing.1
1. C.R. Vol LXXXIX, p. 573.
When I had led M. Pasteur to acknowledge the presence of the microzymas in the altered blood, I was anxious to make him confess the presence of bacteria in the interior of the gamey mass of flesh in his other experiments. But he refused, saying: "I do not know what you mean in speaking of one of my experiments on flesh."
The excessive role ascribed to the germs of the air by this savant and his pretended demonstration of the imputrescibility of organic matters in general when protected from the germs of the air, have diverted science into a deplorable road. He thus threw doubt upon a truth long since acquired; namely, that all natural organic matters, vegetable and animal, are liable to spontaneous change by a phenomenon of fermentation under the conditions specified by Macquer.
This truth must be re-established if we would understand the real meaning of the experiment of M. Pasteur upon the blood; to do this we must connect it with the introductory matter which precedes the first chapter of this work, which led up to the discovery of the real nature of fibrin, which was 11 ic point of departure for the discovery of the real nature of the blood.
I call to mind then that I proved how a solution of sugar or of any other proximate principle, or their mixtures, were changeable on contact with the air, owing to the ferments horn of the germs of this air. M. Pasteur, who had previously asserted the spontaneous generation of ferments, repeated my experiments and was convinced. Then he generalized and asserted that it would be the same in the case of urine and of milk, resembling in this sweetened yeast broth which, being boiled, was not altered if creosoted or left to itself in calcined air.
Before the experiment on the blood or on the flesh, this celebrated savant had experimented on urine and on milk. As to fresh milk, he admitted, a priori, that it soured owing to ferments born of the germs of the air, and that it was coagulated by the lactic acid which coagulated its casein. But here we have boiled milk coagulating in calcined air without becoming sour, while vibrios appeared in it. He was surprised at this, but did not in anywise seek to fathom the mystery, maintaining that in milk the germs of the air resist heating to 100 ° C. and become vibrios, to which he ascribed the coagulation.1
1. His Memoir should be read (Ann. de Chimie et de Physique, Vol. LXIV, pp. 58-63) to realize the efforts made by M. Pasteur to convince himself that the germs of the air are the sole origin of the vibrios.
I have narrated, in the introduction, how I applied the new method of research to milk and thence to various other tissues; similarly I studied from the point of view of their chemical and anatomical changes urine, birds' eggs, fruits which become over-ripe, sprouted barley, frozen plants after a thaw and globules of beer yeast, etc.
Now for the chemical and anatomical facts regarding milk, upon which I cannot insist too strongly. The first phase of its alteration is the separation of the milk globules in the cream; this separation corresponds in an inverse sense to the separation of the blood globules in the cruor of the blood of the horse; the second phase is the souring which precedes the formation of the clot; and this souring corresponds to a fermentation which produces alcohol, acetic acid and lactic acid, the agents whereof are solely the microzymas proper to the milk, for at the moment when the clot is formed, whether the milk has been creosoted or not, the microscope discloses only these microzymas, which have become more readily visible. The vibrios or bacteria which then appear mark the anatomical phase of the phenomenon. But for the complete understanding of the phenomenon of the alterations which occur in milk, it is necessary, as in the case of the blood, to recognize that after its issue from the gland its anatomical elements are no longer in their normal conditions of existence. Also perfectly fresh milk does not contain lactic acid-contrary to the opinion of Berzelius—but it contains alcohol and acetic acid; it hence results that the production of lactic acid after the milking indicates a functional change in the microzymas of the milk; and this formation of lactic acid taking place without the disengagement of gas, especially of hydrogen, indicates in addition that the microzymas of the milk are different from those of the blood.
And since it relates to the phenomenon of coagulation, and that the clot of milk has been compared to the clot of blood, it must also be recognized that the milk clot is not the coagulation of the casein by lactic acid. The casein, in fact, is an insoluble albuminoid proximate principle, which exists in the milk in the state of a soluble caseinate; the acids, whether it be the lactic or the acetic acid, saturate the alkali and the casein is precipitated; it results from this that that .which is called coagulation of the casein in the milk, which is spontaneously altered, is the slow precipitation of the casein by the acids which render the milk sour.
With regard to the coagulation of boiled milk, where there is no souring, that is a phenomenon of another kind in which the caseinates, the albuminates and the zymas of the milk modified by the heat take part; it is a zymastic action, which may be likened to coagulation by rennet, the zymas whereof has its origin in some functional modification by heat of the microzymas of the milk. And this functional modification of these microzymas is so certain that, if there be added to the milk quantities of creosote or of carbolic acid sufficiently great to prevent the vibrionian evolution of the microzymas, there will be no souring or coagulation of the milk; the albuminoid matters undergo other transformations and at last, if the action continues for a long time, at 30° to 35 ° C., the milk globules are destroyed, the fatty bodies which they contain being set free.
The preceding facts regard especially the milks of the cow and of the goat, which are casein milk.
The milks of the ass and of women do not contain casein, but they sour spontaneously without coagulating and yield no clot on the addition of rennet.1
Normal human urine, creosoted, ferments without disengaging gas, producing alcohol, acetic acid and benzoic acid proceeding from hyppuric acid, while the epithelial cellules are destroyed and the microzymas evolute.2
1. On the histological constitution and the comparative chemical composition of the milks of the cow, of the goat, of the ass and of woman"; "On the spontaneous alterations of milk and on the changes which heating produces in it." (M. Chamalet. Passage de Choisel, 60. Paris.)2. C. R., Vol. LXI. p. 374, and Les Microzymas, etc.. p. 713.
The liver, plunged in carbolized water, produces, with the disengagement of carbonic acid, hydrogen and sulphuretted hydrogen, alcohol, acetic acid, lactic acid, while its cellules are destroyed and its microzymas evolve and become bacteria.1
But the changes of eggs and of beer yeast are specially conclusive. The egg of the bird is an organism whose function is to produce a bird. Donne, by vigorous jolting, destroyed this organism, mixing, in the shell, the yolk with the white and thus produced a kind of alteration which I studied. The egg of an ostrich thus treated, at a temperature of 30°-35° C. (86°-95° F.), fermented and produced so much gas that the internal pressure became sufficiently strong to throw out a small part of the contents, on a hole being made in the shell. The gases set free were carbonic acid, hydrogen and a trace of sulphuretted hydrogen. When the gaseous disengagement ceased there was no longer any sulphuretted hydrogen. All the vitellin globules had disappeared and the microzymas were preserved with their form, without any trace of vibrios or other organized production. All the glucose of the egg had disappeared while the albuminoid matter had been preserved, the soluble being coagulable by heat. The products of fermentation were alcohol, acetic acid and butyric acid, showing that they had been produced from the lactate. Here then was a fermentation strictly defined, where­in the microzymas, like those of the blood, did not undergo vibrionian evolution.2 In order that the vitellin microzymas may evolve other conditions are necessary.
1. C. R, Vol. LXXV. p. 1830. 2. C. R., Vol. LXVII, p. 523.
The case of beer yeast is still more interesting, for it has to do with a living being reduced to a cellule, whose alteration and total destruction will throw a strong light upon those of the blood globule. Suppose an alcoholic fermentation of cane sugar for which a little more yeast had been employed than was needed for the complete fermentation of the sugar. The fermentation being accomplished the yeast will be deposited in the fermented liquor and be preserved there unaltered indefinitely, as if in lethargy, with its form and its properties. This determined, let us take some fresh yeast, as it comes from the brewery, washed in distilled water to purify it from what it has brought away from the vat, and steep it in from three to four times its weight of creosoted distilled water to destroy the influence of germs of the air. In this situation, so different from its normal condition of existence, at the temperature of about 30° C. (86° F.) and without any trace of air, it will for a long time disengage pure carbonic acid, producing at the same time a relatively great amount of alcohol, acetic acid and other products, preserv­ing its form all the time. Evidently it has only been able to produce all these these things at the expense of its own substance, of its contents, since its tegument at first remains whole. And if the process of alteration is allowed to continue, this tegument itself will disappear, its microzymas will become free and vibrios appear.1
The following is the method by which the mechanism of the spontaneous destruction of the cellule of beer yeast can be most easily studied. It is well known that yeast does not cause fecula to ferment. But what is not known is, that it liquefies the starch of fecula and is completely destroyed in producing the liquifaction, leaving of its organism nothing but its microzymas, the soluble part of their content being left in the circumambient medium. The phenomenon lasts a greater or less time according to the quantity of creosote employed to destroy the influence of the germs of the air. If the quantity of creosote is small the microzymas undergo vibrionian evolution, if it is sufficient the microzymas do not evolve.2 But that is not all. Thus studied, the phenomenon of the spontaneous destruction of the cellule of beer yeast has enabled me to confirm the generality of the fact which I had long before observed in studying the microzymian origin of the vibrioniens.
1. For details and developments see C. R,. Vol. LVIII. p. 601; "Sur les fermentations par les ferments organises" (1864).2. Ann. de chimie el de physique. 4th Series. Vol. XXIII. p. 443. and Sur la nature el l'origine des ferments (1871).
While the globule of yeast is being destroyed and its microzymas set free and begin to undergo vibrionian evolution, several phases of this evolution are to be observed, which Estor and I have described from the commencement of our researches upon the liver, etc.,1 namely, at first the microzymas are scarcely altered in their size and form; then microzymas coupled in the form of the figure 8, then chaplets of microzymas of from 3 to 10 and 20 grains, all of the same size; then vibrios properly so called; then bacteria often very large, motile or not; also the amylobacters of Trecul, free or fastened end to end. When the phenomenon is not checked by an addition of creosote or carbolic acid, all these productions maybe seen at the same time in the field of the microscope. Now if without changing any of the conditions of the experiment the observation of it is continued, it will be seen that all the forms other than the single microzymas disappear successively; first the amylobacters disappear; new forms of smaller dimensions appear and disappear in turn, so that in the end there remain only swarms of motile forms scarcely differing from the original microzymas which had evoluted.
1. C. R. Vol. LXVI. p. 421, and p. 859 (1868).
Speaking then in the language of anatomy, we may say that the microzymas become vibrioniens by evolution; the vibrios, the bacteria, the vibrioniens in general, return to the microzymian form by an inverse phenomenon of evolution, the ultimate forms differing in little or nothing from the microzymas, the anatomical element of the cellule.
It is thus directly demonstrated that a yeast globule, a cellule in general, in being destroyed sets its own microzymas free; that these, if the necessary conditions are realized, become vibrioniens by evolution, which, in the same apparent conditions, by an inverse phenomenon reproduce the microzymas.
So that as Estor and I have demonstrated in the development of the embryonic cellules of the fowl, and as I have demonstrated in the case of beer yeast and in the case of cellules which may develop in the mother of vinegar, the microzymas which are the commencement of all cellular and tissue organization are also their end, being, as we have seen, the end even of the bacteria.
Now that which is true of the microzymas of beer yeast is true also of the microzymas of all cellules, of all tissues, both of animals and of plants. And this fact has been confirmed unwittingly even by those who deny the microzymas and who, to avoid naming them, have called them punctiform ferments; a microzyma or microzymas-producers at the beginning, microzymas at the end, such are the beginnings and the ends of a bacterium and of a cellule.
Thus all natural animal and vegetable matters, that is to say, organized as Bichat conceived them and defined their organization, the anatomical elements morphologically definite, are the only things living in them; yes, all these matters, from the highest in organization down to beeryeast, are spontaneously alterable from the moment that they are no longer in the situation of their natural conditions of existence, chemically and anatomically.
In insisting upon their chemical alterations, especially upon the production of alcohol, of acetic acid, of lactic acid and of benzoic acid, with or without the disengagement of carbonic acid gas, etc., I wished to show that these alterations belong to the class of the best known fermentations, which assume a living figured ferment. But even in the spontaneous alteration of beeryeast, alcohol and acetic acid are not the only products formed; I described others in 1864; on further studying these latter I have found succinic acid, a special gummy substance, a ternary, furnishing mucic acid, leucin and tyrosin, nitrogenous compounds whose formation bears witness that the albuminoids of the yeast contribute to the changes; later others have been found equally nitrogenous, etc. In extending these researches upon yeast to the spontaneous alterations of the flesh of the horse and that of fish, these researches have been verified by isolating similar or analogous products.
But since these spontaneous chemical alterations belong to the class of fermentations which presuppose the presence of a figured ferment, what is this ferment? For if the beer yeast which causes sugar to ferment puts in it a part of itself, of its transformed content which is recovered among the products of normal fermentation, it is not destroyed; it remains whole, its tegument preserving to it its form, with its own anatomical element-microzymas. On the other hand, when it produces alcohol spontaneously, without sugar, it alters, and is destroyed, as are destroyed the cellules and the organization of the blood, of flesh, of the liver, etc. It is not then these cellules and these tissues which are the ferments of the spontaneous fermentations. M. Pasteur sought in the altered blood the vibrio born of the germs of the air and, not finding it, concluded that there was neither fermentation nor even chemical alteration in it; there are nevertheless fer­mentations without vibrios and without cellules in which are produced alcohol and acetic acid; in the first phase of the alteration of milk, for instance, and in that of eggs jolted up within the shell. These ferments are precisely the microzymas, often the vibrioniens resulting from their evolution, and microzymas which are the result of the destruction of the latter, for at a given moment, either at the commencement or at the end of the phenomenon, there is in the medium which is altering or of which the alteration is completed no production morphologically defined other than the microzymas of origin or the microzymas resulting from their destruction.
And this is not a gratuitous assertion, for I have experimentally proved that the microzymas of animal origin and those of the yeast are actually the figured ferments which produce, with sugar or fecula, alcohol, acetic acid, lactic acid, and by fermentation of the lactate of chalk, butyric acid. And it is precisely the microzymas of the microzymian molecular granulations of the blood or those of the blood globules which belong to that class.
From all these experiments it results incontestably that the microzymas of living organisms in general, and those of the blood and of the blood globules in particular, are anatomical elements and are themselves figured ferments; that is to say, that they are living and organized in the same manner as it is admitted that yeast is so; as are also the vibrioniens which these microzymas may become by evolution, out of the same organized substance. But the microzymas are living beings of an entirely special order without analogy, on which I have insisted for a long time and again insist upon as crowning the demonstration that the blood is veritably a tissue.
And now what happens when this or any tissue whatever alters? First it is no longer preserved in the state in which it exists and functions in the organism, in coordination, to speak as does Dr. Antoine Gros, in general coordination with the functioning of all the organs and of their tissues; it has then, as we have established for the albuminoid atmosphere of the hematic microzymian molecular granulations and for the coloring matter of the contents of the red globule, to undergo, owing to the change in the conditions of its existence, some chemical change in some of its parts; it is in short that its special anatomical elements change their form and their function to the extent of being destroyed and disappear­ing, leaving the microzymas as the only trace of their existence which, according to circumstances, do or do not undergo vibrionian evolution. And the anatomical change maybe so rapid, as is well known to histologists, that one is obliged to take steps to preserve the integrity of the tissues. In fact, one or two minutes may suffice, after the blood has been shed, to render it impossible to demonstrate the third anatomical element.
We must conclude then that in all the experiments, including those of M. Pasteur, the chemical and anatomical alteration in the blood is the work solely of the microzymas, which, in certain conditions, do not become bacteria. As to the question, to what order the chemical phenomenon belongs, it is now solved; since every chemical transformation of a proximate principle of organic matter, under the influence of a figured ferment, is called fermentation or putrefaction, it is evident that the spontaneous chemical alterations of the blood are the result of a fermentation or of a putrefaction without fetid products.
Assuredly, whenever the experiment on the blood shall be taken up upon a larger scale, even under the conditions of that of M. Pasteur, other products will be discovered besides those which I have pointed out, and among them I should not be surprised if alcohol should be found to be one of them.
It is now evident what is the real meaning of the phenomenon called the spontaneous coagulation of the blood; it is the following: The blood, being a tissue, is necessarily alterable itself, as is well known are all tissues, and as are all natural organic matters, animal or vegetable, that which is called coagulation being only the first phase of its more complete change, which extends to disorganization and to the disappearance of its globules. And the phenomenon in its entirety is the work of the microzymas, which, acting phys­iologically as ferments, effect the chemical transformation of the proximate principles, and thereby the anatomical changes, which end in the disorganization of the tissues and of the cellules.
But, as I have said, the microzymas are living beings of a special order without analogy, as I have shown in other publications, upon which I have promised to insist afresh to give to this work and to its demonstration their highest character of certainty and also to refute new errors upon which I had only touched in a casual manner in a former work.1 This will be the subject of the following chapter.
1. Microzymas et Microbes, etc. M. Chamalet. pub., Paris, 60 Passage de Choiseul, 60.