Harry E. Morton

Puerperal endometritis: A prospective microbiologic study

This study was undertaken to determine the flora in the endometrial cavity of two groups of puerperal patients: 47 afebrile control subjects and 27 with endometritis. A transcervical culturing technique designed to avoid cervical contamination and to preserve aerobes, anaerobes, and mycoplasma was employed, and bacterial colony counts were reported semiquantitatively. No difference was found in the flora of these two groups. Furthermore, the most common pathogenic organisms (in decreasing order of frequency) were peptostreptococci, peptococci, Bacteroides sp., gamma-streptococci, enterococci, beta-streptococci, alpha-streptococci, and E. coli.

Production of anti-Mycoplasma (PPLO) antibodies in rabbits.

Summary The antibody response is usually not great when rabbits are injected IV with PPLO or injected IM with the PPLO mixed with adjuvant. The response is better if a primary IM injection of the organisms with adjuvant is followed in a few weeks by one or more IV injections of the organisms. The production of agglutinins is greater following the injection of small amounts of organisms with adjuvant into the hind foot pads and often the response is very good within a period of 7 days. Growth-inhibiting properties have appeared in the sera in a period as short as 14 days following FP injection. Due to the difference in response to FP and IM injections of antigen, injections by both routes may be made at the same time and thus obtain both immediate and prolonged antibody production and sera of higher titer than by either route singly. Intracorneal injection offers a route by which it is possible to obtain a good antigenic stimulus without the use of an adjuvant. Otherwise it appears that the use of an adjuvant is necessary with PPLO for good antibody production. Either Freunds complete adjuvant or the sodium alginate-calcium gluconate adjuvant is satisfactory. Use of the latter adjuvant does not introduce antigens other than those of the Mycoplasma.

Reversible inhibition of mitosis in lymphocyte cultures by non-viable Mycoplasma.

Summary Sufficiently concentrated non-viable Mycoplasma hominis type I suspensions will inhibit the mitogenic action of phytohemagglutin in lymphocyte cultures. The inhibition can be reversed by removing the PPLO as late as 48 hours after inoculation. Mitosis can be started with PHA, stopped by PPLO, and restarted without additional PHA by changing the medium.

Detection and Possible Source of Contaminating Pleuropneumonialike Organisms (PPLO) in Cultures of Tissue Cells

Summary 1) A total of 37 tissue cell lines growing in the presence of horse, calf, human, rabbit or dog serum have been tested for contamination by PPLO. Of these, 22 or 59% contained PPLO. Human, monkey, and mouse cell lines were contaminated. All first and second passages of cell lines were negative. PPLO were isolated from tissue cells growing in human, horse or calf serum. Nineteen positive cell lines were, at some time, grown in presence of human serum. Three positive lines growing in horse serum had never been in contact with human serum. 2) All 25 samples of sera, including pooled lots of human, calf, and horse sera, were negative for PPLO. 3) Possible sources of PPLO contamination of tissue cell cultures are discussed.

Further characterization of the protein factor required by certain pleuropneumonia-like organisms for growth in vitro

Abstract A protein growth factor required by pleuropneumonia-like organisms for growth in vitro can be separated into a partially purified fraction (fraction B). This is composed of two electrophoretically distinct components. Chromatographic separation of these components on alumina columns yielded further purified products, only one of which (fraction B-II) is required by the strains of pleuropneumonia-like organisms tested. The amino acid content and the nature of the low-molecular-weight protein are described.

Electron Microscopic Studies of Biological Reactions. I. Reduction of Potassium Tellurite by Corynebacterium diphtheriae.

Summary (1) Typical polar granules appear as dense spherical masses, or possibly plates, in electron micrographs of unstained preparations of C. diphtheriae grown on blood infusion agar. (2) In addition to polar granules, needle-like crystals appear in electron micrographs of unstained preparations of C. diphtheriae cells grown on potassium tellurite chocolate agar. (3) The needle-like crystals, as well as the black color, of cell masses of C. diphtheriae grown on potassium tellurite chocolate agar disappear upon the addition of small amounts of bromine water. It is inferred, therefore, that the black color is due to the tellurium metal which occurs in the form of needles. (4) It is to be further inferred that the tellurite ion is able to diffuse through the cell wall and is reduced with the precipitation of tellurium metal within the cell boundaries. (5) With the aid of the electron microscope it is now possible to obtain pictorial records of the location of sites of certain chemical reactions incident to the metabolism of the bacterial cell.

Conversion of pleuropneumonialike organisms to bacteria.

Summary A high frequency of appearance of diphtheroids in liquid culture of 3 established strains of pleuropneumonialike organisms has been noted. Statistical, morphological, biochemical and serological tests have shown that a relationship exists between these diphtheroids and the pleuropneumonialike organisms in which cultures the bacteria appear. All PPLO strains tested are serologically related. PPLO strains not giving rise to diphtheroids do not cross react with diphtheroids. Diphtheroids appearing in PPLO cultures differ from oral and air diphtheroids.

Susceptibility of Syrian Hamsters to Leptospirosis

Packchanian 1 studied the susceptibility of 32 species and subspecies of rodents to virulent strains of Leptospira icterohemorrhagiae and concluded that certain species of American deer-mice, genus Peromyscus, are suitable as susceptible small laboratory animals for experimental studies of icterohemorrhagic spirochetosis and for the diagnosis of Weils disease. He did not, however, include hamsters in his studies. Larson 2 reported that white mice (Mus musculus) are extremely sensitive to L.icterohemorrhagiae. He emphasized that young animals be employed. The mortality rate approximates 100% in 3-week-old mice, but falls rapidly as age increases. At this time we wish to report on the susceptibility of Syrian hamsters to L. canicola and L. icterohemorrhagiae. The Syrian hamsters were obtained originally from H. C. Schaefer, Research Department of Ralston Purina Co., St. Louis. The strain of L. canicola was obtained from Dr. Karl F. Meyer, George Williams Hooper Foundation, San Francisco. It was isolated from dog blood by culture methods. The strain of L. icterohemorrhagiae was obtained from Dr. Ruth E. Miller, Womans Medical College of Pennsylvania, Philadelphia. It was isolated from rats in Philadelphia. The culture medium was Schüffners modification of Verwoorts medium, as described by Meyer, Stewart-Anderson, and Eddie. 3 L. icterohemorrhagiae: 0.5 ml of cultures which had been incubated approximately one week at room temperature, when injected subcutaneously into hamsters 3 to 5 weeks old, produced death in 5 to 8 days. Cultures made from the hearts blood 24 hours after the injection remained sterile, but those made 48 and 72 hours after the injection were positive for leptospira. Animals not over 5 weeks old should be used if it is desired to produce death of the animals with typical sign of icterus. L. canicola: 0.5 ml of cultures which had been incubated approximately one week at room temperature, when injected subcutaneously into hamsters 3 to 5 weeks old failed to bring about death of the animals.

Susceptibilities of Pleuropneumonia-like Organisms to some Selective Bacteriostatic Agents.

Summary Strikingly, the PPLO do not behave either as gram-positive or gram-negative microorganisms in that they are not inhibited on solid medium by a 1:250,000 dilution of crystal violet which normally inhibits gram-positive microorganisms nor by a 1:50,- 000 dilution of potassium tellurite which normally inhibits gram-negative organisms. By being able to inhibit both gram-positive and gram-negative microorganisms the isolation of PPLO from mixed cultures can be greatly facilitated. Generally, strains which have been cultivated on artificial media for some time are more resistant to the bacteriostatic substances than are the recently isolated strains.

Symbiotic Growth of Pleuropneumonia-iike Organisms with Bacterial Colonies

Summary Nine of 14 strains of PPLO of human origin and one strain of rat origin were observed to grow on beef heart infusion agar without ascitic fluid or serum only in symbiosis with Staphylococcus albus and all 15 strains grew in symbiosis with Proteus vulgaris.