Sterilization of Crystalline Media and Living Media.—Ostwald varied these experiments by using salol. He melted the substance by heating it above 39∙5°C.; then, protecting it from crystals of any kind, he let the solution stand in a closed tube. The salol remained liquid indefinitely—until it was touched with a platinum wire that had been in contact with solid salol—i.e., until a crystalline germ was introduced. But if the platinum wire has been previously sterilized by passing it, as the bacteriologists do, through a flame, it can then be introduced into the liquor with impunity.
The Dimensions of Crystalline Germs Comparable to those of Microbes.—We may dilute the solid salol with inert powder—lactin, for example—dilute the first mixture with a second, the second with a third, and so on; then, throwing into the solution of surfused salol a tenth of a milligram from one of these various mixtures, we find that the production of crystals will not take place if the fragment thrown in weighs less than a millionth of a milligram, or measures less than ten thousandths of a millimetre in length. It would seem, then, that these are the dimensions of the crystalline particle or crystallographic molecule of salol. In the same way Ostwald satisfied himself that the crystalline germ of hyposulphite of soda weighs about a thousand-millionth of a milligram, and measures a thousandth of a millimetre; that of chlorate of soda weighs a ten-millionth of a milligram. These dimensions are entirely comparable with those of microbes.
All these phenomena have been studied with a detail into which it is impossible to enter here, and which clearly shows more and more intimate analogies between the formation of crystals and the generation of micro-organisms.
Extension and Propagation of Crystallization. Optimum Temperature of Incubation.—Crystallization which has commenced around a germ is propagated more or less rapidly, and ends by invading the whole of the liquor.
The rapidity of this movement of extension depends upon the conditions of the medium, especially upon its temperature. This is shown very well by Tammann’s experiments with betol. This body, the salicylic ester of naphthol, fuses at 96° C. If it is melted in tubes sealed at a temperature of 100° C., it may be cooled to lower and lower temperatures—to + 70°, to + 25°, to + 10°, to-5° without solidifying. Let us suppose that by some combination of circumstances a few centres of crystallization—that is to say, of crystalline germs—have appeared in the solution. Solidification will extend slowly at the ordinary temperature, at 20° to 25° and thereabouts. On the other hand, it will be propagated with great rapidity if the liquor is kept at about 70°. This point—70°—is the thermal optimum for the propagation of germs. It is the most favourable temperature for what may be called their incubation. As soon as the germs find themselves in a liquor at 70° they increase, multiply, and show that they are in the best conditions for growth.
Spontaneous Generation of Crystals. Optimum Temperature for the Appearance of Germs.—If we consider various supersaturated solutions or liquids in superfusion, we shall soon discover that they can be arranged in two categories. Some remain indefinitely liquid under given conditions unless a crystalline germ is introduced into them. Others solidify spontaneously without artificial intervention, and such crystallization may even be propagated very rapidly under determinate conditions. This implies that these are conditions favouring the spontaneous appearance of germs.
This distinction between substances of crystalline generation by filiation and substances of spontaneous crystalline generation is not specific. The same substance may present the two methods of generation according to the conditions in which it is placed. Betol furnishes a good example of this. Liquefy it at 100° in a sealed tube and keep it by means of a stove above 30°, and it will remain liquid almost indefinitely. On the other hand, lower its temperature and leave it for one or two minutes at 10°, and germs will appear in the liquor; prolong the exposure to this degree of heat and the number of these spontaneously appearing germs, appearing in isolation, will rapidly increase. On the other hand, you will observe that propagation by filiation—that is to say, by extension from one to another—is almost absent. The temperature of 10° is not favourable to that method of generation; and we have just seen, in fact, that it is at a temperature of about 70° that extension of crystallization from one to another is best accomplished. The temperature of 70° was the optimum for propagation by filiation. Inversely, the temperature of 10° is the optimum for spontaneous generation. Above and below this optimum the action is slower. We may count the centres of crystallization, which slowly extend further and further, as in a microbic culture one counts the colonies corresponding to the germs primitively formed. To sum up, if there is an optimum for the formation of crystals, there is a different optimum for their rapid extension.
The Metastable and Labile Zones.—This phenomena is general. There is for each substance a set of conditions (temperature, degree of concentration, volume of the solution) in which the crystalline individuals can be produced only by germs or by filiation. This is what occurs for betol above the temperature of 30°. The body is then in what Ostwald has called a metastable zone. There is, however, for the same body another set of circumstances more or less complete, in which its gems appear simultaneously. This is what happens for betol at about the temperature of 10°. These circumstances are those of the labile zone or zone of spontaneous generation.
Crystals of Glycerine.—We may go a step further. Let us suppose, with L. Errera, that we have a liquid in a state of metastable equilibrium, whose labile equilibrium is as yet unknown. This is what actually occurs for a very widely known body, glycerine. We do not know under what conditions glycerine crystallizes spontaneously. If we cool it, it becomes viscous; we cannot obtain its crystals in that way. It was not found in crystals until 1867. In that year, in a cask sent from Vienna to London during winter, crystallised glycerine was found, and Crookes showed these crystals to the Chemical Society of London. What circumstances had determined their formation? We knew not then, and we know not now. It may be observed that this case of spontaneous generation of the crystals of glycerine has not remained the solitary instance. M. Henninger has noted the accidental formation of glycerine crystals in a manufactory in St. Denis.
It may be remarked that this crystalline species appeared, as living species may have done, at a given moment in an environment in which a favourable chance combined the necessary conditions for its production. It is also quite comparable to the creation of a living species; for having once appeared we have been able to perpetuate it. The crystalline individuals of 1867 have had a posterity. They have been sown in glycerine in a state of superfusion, and there they reproduced themselves. These generations have been sufficiently numerous to spread the species throughout a great part of Europe. M. Hoogewerf showed a great flask full to the Dutch biologists who met at Utrecht in 1891. M. L. Errera presented others in June 1899, to the Society of Medical and Natural Sciences at Brussels. To-day the great manufactory of Sarg & Co., of Vienna, is engaged in their production on a large scale for industrial purposes.