3. Evidence for the Vesicular Nature of the Flowing Amebae
The first hypothesis, that the flowing amebae were vesicles enclosed in a membrane, was formulated about two years ago. However, the study of earth bions largely deflected us from the questions surrounding the amebae. Evidence for this hypothesis was not forthcoming. Controlled microscopic observations were lacking. I now hoped for a time-lapse film apparatus that would disclose the process on film. However, for a whole year, the problem could not be solved because of technical difficulties. Time-lapse photography requires a totally motionless preparation and an unstirred solution. It soon became obvious that grass infusions die from lack of oxygen when enclosed in a paraffin. However, a film of the membrane formation around the vesicle and the subsequent development of amebae from those structures could constitute the major evidence.
The technical problem was solved in the following manner: Two to three pieces of moss or grass, clearly separated from each other, are placed on a hanging drop slide, and water for the preparation is added. A piece of was is put under each of the four corners of a cover slip, which is then placed over [the slide] in such a way that no air bubbles form. The cover slip is now attached firmly with paraffin to the two longer sides of the slide. In doing so, one has to make sure that approximately one fourth of the slide cavity is not covered by the cover slip. Water is then pooled along the two sides of the cover slip not covered with paraffin so that the evaporating solution can be replaced continuously with water from the two reservoirs set up o either side. About every two hours, the two reservoirs have to be replenished with water. In this way, the technical problem is solved, and time-lapse photography is made possible over several days; however, some effort and attention are necessary so that there is no delay in refilling the reservoirs lest the preparation dry up in the meantime. With the aid of these technical considerations, dried moss in water was shown to produce amebae, in addition to other protozoa, under conditions not yet fully understood. Thin fragments of grass, which have been made to swell, disintegrate into the most delicate vesicles. One can then observe them growing larger and [note] a distinct membrane being formed around aggregates of vesicles which are initially spherical. When the membrane has reached a certain size, here or there it begins to protrude, but the whole structure us still motionless. A small, round circle appears at the center. Then, a loose outer layer is formed around the large membrane-enclosed structures that have at their center a small, almost punctate vesicle. This will later become the ectoplasm, while the vesicle within the large, round structure give rise to the coarse, vesicular quality of endoplasm.
After several hours, the structure becomes detached from the fiber and continues to move like an ameba. Only in the very last stage of this process does streaming occur in the protruding membrane. The detachment takes place gradually. When the structure has broken free from the fiber, one can already see pseudopodia in various parts of the organism. In this way there is a continual development of amebae. Sometimes, there are two or three simultaneously forming within a short period of time; at other times, they form at specific intervals. The detached amebae divide. Often, they are found in colonies. In this way, they can hardly be distinguished from the margins along moss fibers which have undergone swelling. Evidence for the formation of amebae from swelling moss fibers was provides by this direct observation. (Compare the microphotographs from the time-lapse film, Figures 26-31.)
Figure 26. Moss beginning to swell, 300x
Figure 27. Moss showing vesicular disintegration. Dark Field, about 800x.
Figure 28. A phase in the development of flowing amebae. The vesicles directly above are from swollen moss.
Figure 29. A further stage of the development of the ameba in the same preparation. The vesicles on the left are in the process of dispersing. 1000x
Figure 30. Fully developed amebae wandering off. (Directly over the dispersing vesicles.) 800x.
Figure 31. Vesicular structure of an ameba. 1500x
4. Motile, Vesicular Earth Crystals and Earth Bions
My observations and consequent postulates were at variance with the "germ theory." I purposely avoided orienting myself anew on the prevalent views of the "origin of life from life germs." After these first observations, more than ever, a totally unbiased approach to the work was necessary. For comparative purposes, I set up several unsterile infusions consisting of a tulip leaf, a rose petal, grass, and plain earth. After three days, the tulip leaf and rose petal infusions did not show any protozoa; the grass infusion was full of motile rods, vesicles, and various Protista; the earth infusion (Prep. 32) was a big surprise.
Immediately after the preparation of the infusion, microscopic observation showed distinct membrane-enclosed, unorganized crystals that were completely motionless (Figure 32).
Figure 32. Unorganized earth crystal. Film prep. 1,2,3 - 800x.
Scattered among the crystals, one could find vibrating vesicular structures. Here and there bright green, elongated rods without nuclei were slowly moving around. On the third day, the same open preparation looked quite different. It was full of motile structures which, though sharply angular, moved in exactly the same fashion as the rods and vesicles. It was noticeable that the small vesicular structures often adhered to the surfaces of rods or larger structures. The structures had a taut, vesicular organization. Many of them had changed in their organization; striations had appeared inside, which here and there had undergone vesicular disintegration. The vesicular configurations within the organized crystals could not be distinguished from the vibrating, freely moving vesicular structures in the solution.
On the seventh day, the vesicular disintegration and organization had progressed extensively. At a magnification of only 700x, one could observe clearly delineated protrusions along the edged of the angular, irregular, brow-colored clumps of earth, which looked like tubular vesicles alternately expanding and contracting; angulation was also observed. At a magnification of 1625x, I observed a brown clump whose margins had protruded here and there like vesicles. It was connected to another clump by a vesicular, striated, organized substance. At the junction the clump was bending and elongating. At first, I thought I had been mistaken, but continues careful observation left no doubt: The clump of earth moved as if it had joints, expanding and contracting.
Figure 33. Crystal showing the beginning of vesicular organization. Six week old prep. 1a, 1000x.
Figure 34. Crawling crystal showing vesicular organization.
Figure 35. Moving pseudo-ameba. Sterile earth bound together by gelatin.
After another seven days, the process of vesicular disintegration and striated organization had progressed significantly, as had the vesicular disintegration of the crystal edges. The vesicular protrusions along the crystal edges showed three kinds of movement:
Rotation about the axis;
Expansion and contraction;
Angulation
Let us call the newly described structures on the crystals "plasmoids" (Figures 36, 37).
I passed a 1 mA current through, which I very gradually increased to 2 mA. The movements increased significantly: The vibrating motion of the vesicles became more rapid; expansion and contraction more distinct. When the current was turned on, the vesicles moved toward the cathode. Therefore, they were positively charged particles. When the current was turned off, this movement stopped as promptly as it had started when the current was applied. When the current was reversed, the direction of movement of the individual structures was quickly reversed. When current was sent through for a prolonged period of time, the swollen tubular structures along the edges attempted to break loose in a jerky fashion. Occasionally, these attempts continued for a short while after the current had been turned off. Often, the tubular structures doubled in length. Repeated checking of their reaction to electricity always gave the same result. Therefore, the electric current had some influence on the spontaneous movement, especially elongation, that could not yet be clearly defined.
I will return later to the above reactions, and focus now on some other phenomena observed during the subsequent course of my work.
Let us note that, so far, we have observed two phenomena, which are-in all likelihood-fundamentally different:
1. the vesicular disintegration of swelling plant fibers, i.e., organic tissues, and
2. the vesicular organization of earth crystals, i.e., inorganic material, with subsequent formation of motile, tubular structures and other motile particles.
From then on, I attempted to reproduce the two phenomena experimentally over an over again and to ramify the experimental setup. Above all, I wanted to verify my hypothesis that the vesicles formed from the swelling of substances were actually identical to the vesicular structures within the amebae.
I henceforth replaced the water in the preparation with 0.1 N KCl to test the effect of potassium on swelling. I found that earth treated with KCl began to swell more rapidly and the described phenomena appeared more distinctly; a similar result was obtained when completely- or half-dried grass was treated with KCl. In the grass infusions, I could regularly elicit the production of motionless, vesicular heaps. I became more and more accustomed to seeing the individual vesicles streaming in and out. It was also possible to reproduce over and over again the progressive vesicular disintegration of the earth preparations treated with KCl.
I was interested in artificially coalescing the vesicles formed as a result of the disintegration. I therefore added very dilute red gelatin to the grass and earth preparations that were in a stated of advance disintegration. After only a few hours, one could observe ameboid structures that had not existed previously and that were not present in the preparations lacking gelatin. It took about one to two days for these structures, which I dubbed "pseudo-amebae," to develop fully. They were heaps of vesicles which moved around jerkily in different directions. Occasionally they had individual rods protruding form their membranes like motile spikes. The movements were jerky, not organically flowing like cytoplasmic streamings of real undulating amebae. Aside from locomotion, movements inside the structures, as well as contraction and expansion, could be clearly seen at a magnification of approximately 2000x. As I had postulated, the gelatin had consolidated a number of individual vesicles into heaps of vesicles. The heap of vesicles now continued to move as a unit, the next problem was to determine how this movement originated. Flowing cytoplasmic streaming, observable in many amebae, could not be seen in these preparations (Figure 35).
Figure 36. Plasmoid earth crystal, motile protrusions along edges, 1650x.
Figure 37. Motile plasmoid earth crystal.
Figure 34 shows a crystal completely organized into vesicles and connected by a stalk to a second crystal (organized only along it left margin). At the time the photograph was taken, this crystal moved very slowly in the direction of the arrow, pulling the partially structured crystal behind it.
Now, one could argue that the pseudo-amebae described above had not been assembled artificially, but that "germs of living organisms" had invaded the unheated, open preparation. In order to test the validity of this objection, I now began to heat the preparation in sealed glassware for a quarter to half an hour.
This trial brought a totally unexpected result: It became apparent that the heated infusions displayed far more abundant an active life immediately after their preparation that did the unheated ones after days of swelling.
In the unheated preparations, the vesicles were scattered individually; unorganized crystals were predominant; the vesicular organization of the crystals developed slowly over weeks. The heated earth preparations, on the other hand, showed-immediately after heating-a vast number of tightly packed, active, continually moving, spherical as well as irregular-shaped, vesicles. In the unheated preparation, the solution usually remained clear and contained sediment. In the heated preparation, it immediately clouded up and became colloidal. Electrical testing of the heated preparation showed that the vesicles were also positively charged particles that moved toward the cathode when a current of 0.5 and 1 mA was passed through. On subsequent addition of diluted sterile gelatin, the same result was obtained as earlier with the unheated earth: the aggregation of Individual granule-vesicles into motile ameboid structures. The same result obtained when we heated a mixture of earth, KCl, and gelatin.
However, I was greatly puzzled by the presence of motile structures in a heated, tightly closed preparation immediately after heating. How was it possible that the heated preparation showed much more life than the unheated, unsterile, open one? This seemed to contradict all laws of sterilization. Six days after heating, a tightly closed preparation showed, at a magnification of approximately 1500x, that most of the structures exhibited the familiar movements. There were also plasmatic protrusions and formations, brightly luminating spots with three or four threadlike or rodlike extensions.
Most of the crystals were completely composed of vesicles. Corresponding controls with fresh, unheated earth and water preparations showed the same result over and over again, namely, a paucity of vesicular structure and a dearth of vesicles lacking a nucleus. In my observations, I had to learn to distinguish the spontaneous movement of clumps of earth that had undergone swelling from the passive movement of those recoiling from collisions with motile structures. I could no longer have any doubts about the far more lively nature of the heated preparations; nor the incomparably more abundant formation of contractile sacs and swelling along the edges.
With the use of strong lenses and water immersion and a magnification of approximately 2300x to 3000x, I could clearly observe pulsation in separate parts of the contractile sacs. Solidly crystallized structures with organized margins, which seemed to be connected to one another to one another by a gelatinous substance, attempted to "break free." I was surprised to see, though only at the highest magnification, green, luminous highly motile formations within the gelatinous substance, adhering to some of the structures and indistinguishable from the green, luminous, nucleus-containing rods floating freely in solution. This led to the assumption that the clumps of earth underwent swelling and developed into vesicles or rods in certain places. There are as yet unknown factors involved that determine whether these swollen units remain inside the crystal or break looses and float freely in the solution. When an electric current of 0.5 to 1 mA was passed through, these preparations also showed the now familiar phenomena of vesicular protrusions, increased vibration of the vesicle against each other, movement toward the cathode, etc.
It became increasingly clear to me: The more vesicular the structure of the crystal, the more motile it becomes. In order to solidify my future work, I had to postulate that the vesicular components are swollen units of matter of high electric charge, which are held together by a gel-like substance, and which move within the ameboid structures just as they do in their free state as separate vesicles without a nucleus.
Repeated control experiments with heated earth gave the same results over and over again: motile vesicles, structures with and without a nucleus, pseudo-amebae, and divisions.
In order to rule out faulty preparations as the source of these phenomena, I set up control infusions of unheated earth, as well as tree and tulip leaves, etc., which I left exposed to the air. Over and over again, the above described result was obtained, namely, that the unsterile preparations shoed far less vesicles than the heated ones, and that it was far more difficult to obtain marginal layers that had become motile or had undergone vesicular disintegration.
The "process of ingestion" drawn from life.
Ameba A did not move toward bions B; however, after a certain distance, the bions moved rapidly toward the ameba and disappeared inside (note direction of arrow)
The question now arose whether these structures, if they actually represented living matter, showed, aside from movement, contraction and expansion, still other known properties [of the living], such as cell division. Therefore, I began to study the earth-KCI-gelatin preparations very carefully. Within a short space of time, after focussing for hours on a single field or structure, I could observe pseudo-amebae divide.
The more familiar I became with the preparations, the more probable it became that I was dealing with living organisms which, however, lacked completeness, so to speak; for example, the movements of the pseudo-amebae were jerky, sluggish, vibrating, without inner streaming, "mechanical." The structures had to be preliminary forms of living organisms. To begin with, for my own personal use, I called them bions. Could full life develop from these bions?
It was very satisfying to detect the function of "ingestion," in addition to cell division. I was able to observe that such pseudoameboid heaps of vesicles ingested individual vesicles that were freely moving about. I thus had to free myself of an idea that is erroneously connected with the word "ingesting." No doubt, we make the mistake of using anthropomorphic expressions when describing protozoal structures. When we say "ingesting," we automatically think that a "rational" living organism "takes in" something from its environment "in order to maintain its life." Only after I had freed myself from this erroneous thinking was I able to incorporate the phenomenon of "ingesting" logically into my other observations: To begin with, it seemed inevitable that the cells I had in front of me were composed of vesicles and rods, just as a multicellular body is composed of individual cells. This contradicted the view that cells are the basic units of living matter; because then it would not be the cells but the vesicles that would constitute a biological unit, and the cells would already be complex structures. Yet, these vesicles and rods, as shown by their reaction to electricity, were highly charged, tautly stretched structures. Hence the obvious assumption that a heap of vesicles carries out its "process of ingesting" by attracting individual vesicles which are free-floating and electrically charged, and that it incorporates them into the aggregate it is forming, just as we had observed with the contractile protozoa.
This was simply a hypothesis that came to me. I would have gladly accepted any other workable one. At this stage in my work, further development of the experiments and their controls seemed to me most important. Over and over again I came up against the question as to how it was possible that heated substances showed more life than unheated, unsterile ones. Originally I thought that the rapid, vibratory movement of the vesicles after heating was due to heat. However, several tests with electric current showed this movement to be of an electrical nature, since this vibratory motion also increased when current was sent through. I did not yet want to consider whether I was dealing here with actual bacteria or cocci, or whether these were structures of a different nature that had been formed a s result of heating. Since it had become possible to produce pseudoamebae from heated earth, gelatin, and KCI, it seemed feasible to perform a further control experiment with other substances. According to my hypothesis, there had to be substances that removed the "incomplete" character from the structures.
Later, I proceeded to autoclave the earth in KCI at 120 degrees. The result was much better as compared to the merely heated coal (at 180 degrees), i.e., there were a greater number of more organized crystals, and the vesicles and heaps of vesicles were almost completely motile. I will return to this when I describe my attempts to culture them.
The observations described so far confirm the biological correctness of the tension-charge formula. The vesicular swelling represented mechanical tension; the occurrence of electrical positivity or negativity on the individual vesicles represented electrical charge. However, most of this still remained in the dark.
© 2008 The American College of Orgonomy. All rights reserved.
