the production of a change in the corresponding bacterium whereby it becomes granular, swells up and ultimately may (a) Lyso-
genic action. undergo dissolution. Pfeiffer was the first to show that this occurred when the bacterium was injected into the peritoneal cavity of the animal immunized against it, and also when a little of the serum of such an animal was injected with the bacterium into the peritoneum of a fresh, i.e. non-immunized animal. Metchnikoff and Bordet subsequently devised means by which a similar change could be produced in vitro, and analysed the conditions necessary for its occurrence. It has been completely established that in this phenomenon of lysogenesis there are two substances concerned, one specially developed or developed in excess, and the other present in normal serum. The former (Immunkörper of Ehrlich, substance sensibilisatrice of Bordet) is the more stable, resisting a temperature of 60° C., and though giving the specific character to the reaction cannot act alone. The latter is ferment-like and much more labile than the former, being readily destroyed at 60° C. It may be added that the protective power is not lost by exposure to the temperature mentioned, this apparently depending upon a specific anti-substance. Furthermore, lysogenic action is not confined to the case of bacteria but obtains also with other organized structures, e.g. red corpuscles (Bordet, Ehrlich and Morgenroth), leucocytes and spermatozoa (Metchnikoff). That is to say, if an animal be treated with injections of these bodies, its serum acquires the power of dissolving or of producing some disintegrative effect in them. The development of the immune body with specific combining affinity thus presents an analogy to antitoxin production, the difference being that in lysogenesis another substance is necessary to complete the process. It can be shown that in many cases when bacteria are injected the serum of the treated animal has no bacteriolytic effect, and still an immune body is present, which leads to the fixation of complement; in this case bacteriolysis does not occur, because the organism is not susceptible to the action of the complement. In all cases the important action is the binding of complement to the bacterium by means of the corresponding immune body; whether or not death of the bacterium occurs, will depend upon its susceptibility to the action of the particular complement, the latter acting like a toxin or digestive ferment. It is to be noted that in the process of immunization complement does not increase in amount; accordingly the immune serum comes to contain immune body much in excess of the amount of complement necessary to complete its action. An important point with regard to the therapeutic application of an anti-bacterial serum, is that when the serum is kept in vitro the complement rapidly disappears, and accordingly the complement necessary for the production of the bactericidal action must be supplied by the blood of the patient treated. This latter complement may not suit the immune body, that is, may not be fixed to the bacterium by means of it, or if the latter event does occur, may fail to bring about the death of the bacteria. These circumstances serve, in part at least, to explain the fact that the success attending the use of anti-bacterial sera has been much inferior to that in the case of antitoxic sera.
Another property which may be possessed by an anti-bacterial serum is that of agglutination. By this is meant the aggregation into clumps of the bacteria uniformly distributed in an indifferent fluid; if the bacterium is motile (b) Agglu-
tination.its movement is arrested during the process. The process is of course observed by means of the microscope, but the clumps soon settle in the fluid and ultimately form a sediment, leaving the upper part clear. This change, visible to the naked eye, is called sedimentation. B. J. A. Charrin and G. E. H. Roger first showed in the case of B. pyocyaneus that when a small quantity of the homologous serum (i.e. the serum of an animal immunized against the bacterium) was added to a fluid culture of this bacillus, growth formed a sediment instead of a uniform turbidity. Gruber and Durham showed that sedimentation occurred when a small quantity of the homologous serum was added to an emulsion of the bacterium in a small test-tube, and found that this obtained in all cases where Pfeiffer’s lysogenic action could be demonstrated. Shortly afterwards Widal and also Grünbaum showed that the serum of patients suffering from typhoid fever, even at an early stage of the disease, agglutinated the typhoid bacillus—a fact which laid the foundation of serum diagnosis. A similar phenomenon has been demonstrated in the case of Malta fever, cholera, plague, infection with B. coli, “meat-poisoning” due to Gärtner’s bacillus, and various other infections. As regards the mode of action of agglutinins, Gruber and Durham considered that it consists in a change in the envelopes of the bacteria, by which they swell up and become adhesive. The view has various facts in its support, but F. Kruse and C. Nicolle have found that if a bacterial culture be filtered germ-free, an agglutinating serum still produces some change in it, so that particles suspended in it become gathered into clumps. E. Duclaux, for this reason, considers that agglutinins are coagulative ferments.
The phenomenon of agglutination depends essentially on the union of molecules in the bacteria—the agglutinogens—with the corresponding agglutinins, but another essential is the presence of a certain amount of salts in the fluid, as it can be shown that when agglutinated masses of bacteria are washed salt-free the clumps become resolved. The fact that agglutinins appear in the body at an early stage in a disease has been taken by some observers as indicating that they have nothing to do with immunity, their development being spoken of as a reaction of infection. This conclusion is not justified, as we must suppose that the process of immunization begins to be developed at an early period in the disease, that it gradually increases, and ultimately results in cure. It should also be stated that agglutinins are used up in the process of agglutination, apparently combining with some element of the bacterial structure. In view of all the facts it must be admitted that the agglutinins and immune bodies are the result of corresponding reactive processes, and are probably related to one another. The development of all antagonistic substances which confer the special character on antimicrobic sera, as well as antitoxins, may be expressed as the formation of bodies with specific combining affinity for the organic substance introduced into the system—toxin, bacterium, red corpuscle, &c., as the case may be. The bacterium, being a complex organic substance, may thus give rise to more than one antagonistic or combining substance.
By opsonic action is meant the effect which a serum has on bacteria in making them more susceptible to phagocytosis by the white corpuscles of the blood (q.v.). Such an effect may be demonstrated outside the body (c) Opsonic action.by making a suitable mixture of (a) a suspension of the particular bacterium, (b) the serum to be tested, and (c) leucocytes of a normal animal or person. The mixture is placed in a thin capillary tube and incubated at 37° C. for half an hour; a film preparation is then made from it on a glass slide, stained by a suitable method and then examined microscopically. The number of bacteria contained within a number of, say fifty, leucocytes can be counted and the average taken. In estimating the opsonic power of the serum in cases of disease a control with normal serum is made at the same time and under precisely the same conditions. The average number of bacteria contained within leucocytes in the case tested, divided by the number given by the normal serum, is called the phagocytic index. Wright and Douglas showed that under these conditions phagocytosis might occur when a small quantity of normal serum was present, whereas it was absent when normal salt solution was substituted for the serum; the latter thus contained substances which made the organisms susceptible to the action of the phagocytosis. They further showed that this substance acted by combining with the organisms and apparently producing some alteration in them; on the other hand it had no direct action on the leucocytes. This opsonin of normal serum is very labile, being rapidly destroyed at 55° C.; that is, a serum heated at this temperature has practically no greater effect in aiding phagocytosis than normal salt solution has. Various observers had previously found that the serum of an animal immunized against
