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I T IS generally accepted that the incidence of antibiotic-resistant bacteria in closed communities is more or less related to widespread use of the drug over an extended period of time. That antibiotic-resistant infections in war wounds likewise should attend routine use of penicillin for prophylaxis of infection would seem logical. Documentary evidence supporting this hypothesis has been obtained in World War II and in Korea. Royet al. 5 made a study of contamination and infection of soft-tissue wounds in 1944-45. They noted an incidence of 17 per cent penicillin-resistant strains of staphylococci in 94 control cases, and an incidence of 49 per cent among 69 penicillin-treated patients. Lindberg and Parrott,2 in a bacteriological study of 25 casualties in Korea, found that two-thirds of the micrococci isolated from their wounds were resistant to penicillin. This communication reports clinical and bacteriological studies in neurosurgical Korean battle casualties. GENERAL PROCEDURE This report is based on 58 consecutive patients treated at the N eurosurgical Center, Tokyo Army Hospital, between February 1951 and May 1952. The majority of these had been wounded in action in Korea, and had received initial surgery by one of the neurosurgical teams or on hospital ships in the Korean Theater, prior to evacuation to Japan. Practically all had received daily intramuscular injections of penicillin and streptomycin from the time of injury to admission. These patients had pyogenic infections of the central nervous system, or in craniectomy or laminectomy wounds. Shortly after admission swabs, needle-puncture aspirates, or tapped spinal fluids were taken from infected areas in every instance. The primary objective was to obtain laboratory guidance in selection of appropriate antimicrobial therapy. In Korea only penicillin and streptomycin were freely available. In our hospital, chloramphenicol (Chloromycetin), also limited stocks of chlortetracycline (Aureomycin) and oxytetracycline (Terramycin), were at hand. For the purpose of obtaining rapid data on sensitivities to antibiotics, the medicated disc-blood agar plate method was established at Tokyo Army Hospital in January 1951 by one of us (E.J.P.).*

The antibiotics, polymyxin A B C D and E, have been described in the recent literature. Their antibacterial spectra are similar. They differ from one another in amino acid content. It is the purpose of this presentation to describe our studies of the antibacterial activity, pharmacology, untoward reactions, and clinical experience with polymyxin B and polymyxin E.5 IN VITRO ACTIONWe found that the susceptibility to polymyxin of 78 strains of ten different genera by the tube dilution test (Table I) is marked against Salmonella, Shigella, Klebsiella, microorganisms of the coli aerogenes groups, and most importantly, Pseudomonas. Brucella and many staphylococci are moderately sensitive, while Proteus and hemolytic streptococci are refractory. It is of interest that a strain of Pseudomonas, which developed high resistance to streptomycin in three daily transfers, did not become resistant to polymyxin after 27 transfers. Size of inoculum and human serum reduced but slightly polymyxin activity. The concentration of polymyxin required to inhibit Klebsiella and Pseudomonas is not reduced significantly by the addition of subinhibitory amounts of streptomycin, aureomycin, sulfadiazine or penicillin, either singly or in several combinations. Polymyxin antagonizes the cumulative streptomycinpenicillin action on Proteus.

The maintenance of a large collection of stock cultures of bacteria for ready availability is a major task. The method of frequent transfer on a suitable culture medium, as commonly employed, requires not only a great expenditure of time, materials and effort, but also involves the possible loss of certain biological, immunological and cultural characteristics; not to mention the 1912). To determine the efficacy of Soyka's method, cultures were grown in beef-infusion broth, pH 7.2, sealed off in ampoules and stored at room temperature in the dark. Such ampoules were opened periodically and their contents transferred to fresh media. The sub-cultures were examined for viability, colony form and morphology. The results thus obtained follow. A. Organisms surviving without change. (a) After 45 months: Bacillus anthracis, Bacillus megatherium, Bacillus mesentericus, Bacillus subtilis, Escherichia coli-communior, Shigella dysenteriae (Shiga and Flexner), Salmonella enteritidis (Stanley), Aerobacter aerogenes, Diplococcus mucosus (types A and B), Eberthella typhosa (4 strains), Proteus mirabilis, Proteus vulgaris, Staphylococcus albums (1 strain) and Staphylococcus aureus (1 strain). (b) After 33 months: Eberthella typhosa "R", Brucella melitensis "R" and Micrococcus aurantiacus. B. Organisms surviving 30 to 45 months with changes as indicated. Escherichia coli "SY) ("R" forms), Escherichia coli "R" ("S" and small colony forms), Shigella dysenteriae (Strong) "S" ("R" forms), Salmonella enteritidis "S" ("R" forms), Salmonella paratyphi A "S" (extreme "R" and small colony forms), 164 on October 4, 2020 by guest http://jb.asm.org/ Downloaded from PRESERVATION OF BACTERIAL CULTURES1 Salmonella pullorum "R" (mucoid borders), Eberthella typhosa "5", 1 strain ("R" forms), Serratia indica and Serratia marcesenw, 4 strains (loss of pigment), Corynebacterium diphtheriae "R" (small colonies), Gaffkya tetragena, 2 strains (small colonies), Neisseria catarrhalis (small colonies), Proteus vulgaris (small colonies), Sarcina lutea (small colonies and loss of pigment), Staphylococcus aureus (aureus and albus type of colonies) and Vibrio proteus (small colonies). C. Organisms not surviving on subculture. (a) After 30 to 45 months: Bacillus novus (Huss), Alcaligenes fecalis, Salmonella paratyphi A, Brucella abortus (bovine and porcine), Chromobacterium violaceum, Corynebacterium hofmannii, Corynebacterium xerose, Micrococcus flavus, roseus and tetragenus, Neisseria catarrhalis, Pasteurella pestis (caviae), Pseudomonas aeruginosa, Sarcina lutea, Staphylococcus albus, Staphylococcus aureus (1 strain), Vibrio metchnikovi and Vibrio schuylkilliensis. (b) After 15 to 23 months: Pseudomonas phosphorescens, Alcaligenes bronchisepticus and S-accharomyces cerevtsiae. Summary. Storage of broth cultures in ampoules is not wholly satisfactory. Some species (the species of Brucella, Pseudomonas phosphorescent, the species of Corynebacterium, Pseudomonas aeruginosa and Saccharomyces cerevisiae) did not survive longer than a few months, and many underwen...