Louis Dienes

Reproductive Processes in Proteus Cultures.

Summary The complex reproductive processes formerly observed in certain Gram-negative bacteria have also been observed in Proteus cultures. In this species they presented several distinctive characteristics. The large bodies in every case previously studied were produced by gradual swelling of the bacteria. In Proteus they are produced by a process known as “plasmoptysis.” The large bodies so produced possess similar physical properties and show reproductive processes similar to those formed in other species by swelling of the bacteria. Another characteristic of Proteus is that the bacteria from which the large bodies develop differ considerably from the bacteria usually present in the culture. Large bodies develop only from spreading filaments actively motile on solid media. In almost every culture these filaments produce an occasional large body, and most of them are transformed into large bodies when the spreading halo of one strain meets the spreading halo of another appropriate strain. The large bodies so produced are viable and within a short time are transformed into regular bacteria. Exceptionally they produce the L type of growth. There is a great variation in the reaction of different strains to each other. The spreading filaments show a tendency to “plasmoptysis” and production of large bodies under various adverse conditions, for example, when they are subjected to refrigeration or when they are transferred into tap-water or into broth containing HgCl. The large bodies produced by refrigeration are apparently fully viable. Those produced by tapwater are only occasionally viable, and when they germinate they usually produce L type colonies. Large bodies produced by HgCl2 are not viable. The difference in viability constitutes an important difference between the large bodies produced by the interaction of the 2 strains and by refrigeration on one hand, and by toxic influences on the other hand.

The role of pleuropneumonia-like organisms in genitourinary and joint diseases.

THE high incidence of acute joint disease in male patients with positive prostatic cultures, in addition to the knowledge that animals infected with L organisms frequently have arthritis, suggests ...

Relationship of Pleuropneumonia-like (L) Organisms to Infections of Human Genital Tract.

In a consecutive series of cultures from the genital tract of 129 unselected patients, pleuropneumonia-like (L) organisms were recovered from 23 of 77 cervical cultures, from 1 of 8 vaginal cultures, from 3 of 36 prostatic cultures, and from 1 of 8 cultures of urethral discharges from males. 1 The 4 male patients who yielded an abundant growth of these organisms all had chronic prostatitis. Gonococci were not demonstrable. One patient had rheumatoid arthritis, one had polyarthritis clinically resembling gonococcal arthritis, one complained of soreness of feet and knees and swelling of fingers. Joint effusions were obtained from the first 2 of these patients, but no organisms could be found in them by smear or culture. Among the females, considering only those cases in which pleuropneumonia-like organisms were the predominating flora and, excluding cases in which only a few colonies were observed, we find that 5 had chronic cervicitis, 7 had gonorrhea, while in 4 the cervix appeared clean. Among the 9 from whom gonococci were not recovered, 1 had rheumatoid arthritis and 3 complained of various skeletal aches and pains. One of these 3 was the wife of the male patient who complained of prostatitis, soreness of feet and knees and swelling of fingers, a finding which suggests the possibility of transfer of the organisms by sexual intercourse. This mode of transmission is further indicated by another finding, not included in the above series. In this case, a man was admitted to the hospital because of acute tenosynovitis of the left wrist and chronic prostatitis.

Further Observations on the Reproduction of Bacilli from Large Bodies in Proteus Cultures.

Summary Single large bodies were isolated from the contact line of 2 spreading Proteus strains. Their descendants were similar to one of the strains with respect to the properties studied, and crossing or segregation of properties was not observed. With a single exception, only one strain was recovered from the large bodies formed at the contact of 2 strains.

Cultivation of Pleuropneumonia-Like Organisms from Female Genital Organs.

The technic formerly described for staining bacterial cultures in situ on the surface of agar was employed in studying routine plates submitted for gonococcus examination. 1 In the course of 2 months pleuropneumonia-like organisms were demonstrated in the cervical secretions of 5 patients. The medium used for the gonococcus is similar to the medium employed formerly in cultivating Streptobacillus moniliformis and pleuropneumonia-like organisms. 1 It is essentially a sedimented boiled blood agar to which is added 30% buffered ascitic fluid. The plates are incubated for 2 days in partial CO2 tension. Pleuropneumonia-like organisms were present in the genitals of about one-third of the females. Thus far similar organisms have not been found in plates inoculated with secretions from the urethra or prostate of males or from eyes of babies suspected of gonococcus infection. However, the female and male material examined was not comparable. The majority of female patients had pelvic infections, while the cultures from males were mostly release cultures from treated gonococcal patients. Women without pelvic disease were not studied. The group of pleuropneumonia-like organisms is characterized at present by purely morphological criteria. The organisms cultivated from the female genitalia are indistinguishable in morphology and in the appearance of colonies from the strains isolated from rats and mice. 2 The young colonies consist of very small pleomorphic granules and filaments which grow into the medium and are stained deeply in situ with methylene blue. The surface of fully developed colonies consists of large bodies (3 to 10 microns) which are at first deeply stained but which later become vacuolized and produce a foam-like structure. After 48 hours the colonies are often only 10 to 20 microns in diameter. In transplant they develop to a considerably larger size.

In vitro sensitivity of some bacteria, their L forms and pleuropneumonia-like organisms to antibiotics.

Summary The sensitivity of PPLO strains of various origin and of several bacteria and their L forms were examined in the presence of 9 antibiotics. The sensitivity of L forms to the various antibiotics with the exception of penicillin was comparable to that of the parent bacterium. However, the L forms of streptococci and of one Vibrio strain were consistently less sensitive to bacitracin than their bacterial forms. All PPLO strains, like the L forms, were highly resistant to penicillin. The sensitivities of the parasitic strains to the various antibiotics were uniform; they were markedly resistant to bacitracin and erythromycin, like some bacteria. The 3 saprophytic strains were noticeably less resistant than the parasitic strains to several antibiotics, especially to erythromycin. The sensitivities of all strains of PPLO, of bacteria and of L forms of these bacteria were on the whole comparable.

Differences between oral and genital strains of human pleuropneumonia-like organisms.

Conclusion and Summary Pleuropneumonia-like strains isolated from the human genitourinary tract and from the buccal cavity, respectively, differ in the appearance of the colonies, in serological properties and, to a certain extent, in nutritional requirements. It is likely that they represent different species. In addition to the easily cultivatable strains, colonies of much smaller size develop occasionally on the plates inoculated from either source. These small colonies retain their identity in transfer. It is uncertain whether they are variants of the usually cultivated strains.

L Type Variant Forms in Cultures of Various Bacteria.

Summary Cultures of various bacteria on blood agar plates were stained in situ by methods previously described. Examination of cultures of a Gram negative bacillus (of the genus Flavobacterium) so stained revealed tiny secondary colonies similar in appearance and morphology to the L1 variant of the Streptobacillus monilliformis. The same phenomenon was observed, in a less pronounced degree, in cultures of Bacillus influensae. Bacillus fundulliformis and in cultures of certain strains of colon bacilli.

Isolation of Pleuropneumonia-like Organisms from H. influenzae with the Aid of Penicillin.∗

Summary With the aid of penicillin a strain of pleuropneumonia-like organism has been isolated from a culture of H. influenzae. This strain could be kept in continuous cultivation. Colonies of similar organisms were seen in 3 other cultures of H. influenzae, but their propagation in pure culture was not successful.

Isolation of L type cultures from clostridia.

Summary L type cultures have been isolated from a strain of Clostridium tetani with the help of penicillin. Similar cultures were obtained from a freshly isolated unidentified Clostridium when the area surrounding a 2-day-old agar culture was reinoculated with the same strain. The metabolic products diffusing from old culture inhibited growth and transformed the bacilli into L forms. The L type cultures obtained are similar in appearance and morphology to those isolated previously from a Gram positive spore-bearing bacillus. They differ from the L forms of Gram negative bacilli inasmuch as they grow better in the absence rather than the presence of animal serum and have a tendency to form large spreading colonies increasing in size for several months.