VARIATION AND TYPE SPECIFICITY IN THE BACTERIAL SPECIES HEMOPHILUS INFLUENZAE
Authors: Pittman Margaret 
Published: June 2004

During the course of a study of different strains of influenza bacilli, fifteen strains have been found to form colonies of a different appearance from that usually considered typical of influenza bacilli. These colonies are smooth, more opaque, and are iridescent in oblique transmitted light. Most of these strains were isolated from patients in whom these organisms seemed to play a pathogenic rô1e. When these strains were grown repeatedly on blood agar, other colonies appeared which were smaller, less smooth, less opaque, and not iridescent, and when subcultures were made from these rough colonies, all of the colonies were of this character. Further study of the cultures obtained from these smooth and rough colonies have shown that one is a variant of the other. The strain from the smooth colony has been called an S strain, that from the rough colony an R strain. Certain differences in the morphology of the organisms in the R and S strains have been observed. Of special importance is the fact that the bacteria of the S strains are possessed of capsules. It has also been found that the S strains are somewhat more virulent for laboratory animals than are the R strains.
In the supernatant fluid of broth cultures of S strains, and in the washing fluid of S bacteria grown on agar, there is present a soluble substance which, in the presence of homologous immune serum, gives rise to a precipitate. No reaction of this kind, however, occurs with the cultures of the R strains. By means of cross precipitin reactions, employing antisera against the different S strains, it has been found that the fifteen strains studied may be divided into two distinct immunological groups. Three of these strains belong in one group, Type a, and the remaining twelve in another group, Type b. Seven of the strains studied were isolated from the spinal fluid in cases of meningitis, and all of these strains are of Type b. Agglutination tests performed at 37°C. with these fifteen S strains have revealed the same specific type relationships among the organisms as did the precipitin tests. The R strains on the other hand, exhibit no similar type agglutinations. If the agglutination tests are made at a higher temperature, 47°C., the S strains also fail to show the specific type reactions which occur at 37°C. Certain differences between other biochemical reactions exhibited by the two types of strains have been noted, but it is not believed that they are sufficiently constant to be of great significance.
When S strains are grown on artificial media outside the animal body, they tend to be converted into the R form. The rapidity and the readiness with which this conversion occurs depend on certain conditions, such as the kind of media employed, the temperature at which the cultures are kept, and the atmospheric conditions under which they are cultivated. The rate of conversion is increased when the S strains are grown in media containing anti-S immune serum of the homologous type. On the other hand, conversion of R strains into the S form occurs with much less readiness, and then only if very particular conditions are present. On one occasion conversion occurred when an R strain was grown in a medium containing anti-R immune serum. On two other occasions this same strain changed from the R to the S form during passage through animals. With other R strains it has so far been impossible to bring about this transformation.
These studies indicate that the bacteria belonging in the group Hemophilus influenzae exhibit changes in pathogenicity and immunological specificity, which are analogous to those shown by the bacteria of the pneumococcus group. It is important to continue this study, with the technique which has been developed, to include a much larger number of strains. On account of the readiness with which the S strains of influenza bacilli lose their type specificity when grown on artificial culture media, it is important that the organisms be studied as soon as possible after removal from their pathological sources. It is not impossible that many strains lose their specificity immediately after removal from the host, and that the specific immunological differentiation of many strains may, for that reason, be very difficult, if not impossible.