Frank L. Meleney

The Identity of C. Oedematoides and B. Sordellii.

In 1927, we, with Karp, 1 reported the isolation from a fatal human case of “gas gangrene” of a pathogenic anaerobic organism which did not correspond either culturally or immunologically to any of the hitherto well recognized clostridia; and, shortly after, we were able to isolate the same organism from specimens of imperfectly sterilized cat-gut. 2 This organism somewhat resembled Weinbergs B. oedematiens culturally. When injected into mice subcutaneously it produced much the same type of lesion, i. e., the white gelatinous oedema so characteristic of the latter organism. It differed from B. oedematiens, however, in failing to ferment glycerol; it was more actively proteolytic; and, more especially, the toxin readily obtained by filtering cultures, was in no way neutralized by high-titre anti-oedematiens serum. On the other hand, anti-serum prepared by the immunization of a rabbit with this sterile filtrate afforded full protection against the homologous toxin, but not at all against the toxins of B. welchii, of vibrion septique or of B. oedematiens. For these reasons we concluded that the organism should be recognized as a distinct species and suggested the name Clostridium oedematoides. A short time later, Hall and Scott 3 published a restudy of 2 strains of organisms sent to them from South America by Sordellii, which the latter had described first in 1922 4 and had named B. oedematis sporogenes. Although Hall and Scott found one of these strains to be entirely non-pathogenic for laboratory animals, they found that the other was fully virulent and that it produced a potent exo-toxin which could be separated from cultures by filtration. Hall and Scott suggested that the original name, B. oedematis sporogenes should be replaced by the binomial B. sordellii. They also suggested that the cultural characteristics of Sordelliis organism were so similar to those of C. oedematoides as described by us, as to render it probable that the 2 species were identical.

Distribution of bacitracin in the body.

Summary One, 2 or 3 hours after a large single intravenous injection of bacitracin, the concentration of the drug in the different tissues of rabbits was found in decreasing order as follows, subject to some individual variation: urine, kidney, blood, bile, lung, bone marrow, skin, large intestine, pancreas, heart muscle, skeletal muscle, liver, spleen, small intestine, cerebrospinal fluid and brain. In the chest and peritoneal exudates produced by aleuronat-starch injection, the titres of bactiracin one hour after an intravenous administration approximated those in the blood.

B. coli Bacteriophage in the Treatment of B. coli Peritonitis in Mice

The bacteriophage which was used for this study was found to produce a complete lysis of 2 virulent strains of B. coli which we obtained from patients with peritonitis. One one hundred billionth of a cc. of this bacteriophage caused lysis of approximately one billion bacteria of strain “T” in 10 cc. of broth. The minimal lethal dose of the “T” strain of B. coli for mice at the time of the experiments was 1-5 million organisms which killed within 5 to 12 hours after injection of an actively growing, 3-hour, 2% dextrose cooked meat medium culture. Fifty million bacteria which constituted 10-50 M.L.D.s were suspended in 0.5 cc. of broth and inoculated intraperitoneally into a series of mice. 0.5 cc. of bacteriophage was injected simultaneously into 2 of these mice, and into 2 more mice at varying intervals up to 4 1/2 hours after the bacterial inoculation. This approached closely the lethal period for control animals. In the control series plain broth injections were given at the same intervals as phage. The table shows one of these experiments. From this table one can see that 0.5 cc. of bacteriophage protected all of the mice when given up to 3 1/2 hours after bacterial inoculation. Only 50% of mice could be saved when bacteriophage was given after 4 hours. In similar experiments there was occasional recovery when phage was inoculated after 4 1/2 hours or later. Control animals receiving B. coli filtrate instead of phage were not saved. The bacteriophage itself proved to be innocuous, for 2 cc. and 4 cc. of it injected respectively into the peritoneal cavity of 2 mice did not produce any harmful effect. Both mice survived and the one inoculated with the larger dose suffered only slight discomfort for 1 or 2 hours, probably due to distention from the quantity of fluid injected.

The viability of hemolytic streptococcus in certain solutions containing gelatin.

For certain quantitative biologic tests it is necessary to use a suspending medium in which the streptococcus will survive without increase or diminution in numbers for a considerable length of time. Falk has recently reviewed the extensive literature on the subject of the inhibitory effect of the cations of various salts on the biologic activities of bacteria and other cells. He concludes that balanced solutions containing two or more salts in the proper proportions preserve the life of bacteria better than either distilled water or solutions containing single salts unless they are very dilute. Robertson, Sia and Woo have recently found that 0.1 to 0.125 per cent gelatin in distilled water or Lockes solution greatly increases the length of life of the pneumococcus in these fluids. With these facts as a basis, experiments with a hemolytic streptococcus obtained from an acute gangrenous infection of the thigh have been carried out with six different fluids: 1, distilled water; 2, “normal” saline; 3, Lockes solution; 4, a balanced solution containing 1 per cent sodium chloride and 0.05 per cent calcium chloride (“20-1”); 5, 0.2 per cent sodium citrate; 6, a balanced solution containing 0.5 per cent sodium chloride, 0.5 per cent potassium chloride, 0.5 per cent calcium chloride and 0.1 per cent magnesium chloride (Zeugs mixture). Each fluid was prepared both with and without 0.1 per cent gelatin and titrated after autoclaving to pH 7.8. The streptococcus was grown in 1 per cent dextrose digest broth, washed twice and then suspended in the solution to be tested at a standard concentration, approximately one billion organisms per cubic centimeter. per cubic centimeter. This was diluted down seven times with the fluid to be tested, each dilution containing a tenth of the number in the previous dilution so that the final dilution was estimated to contain 100 organisms.