In vitro antimicrobial activity of dimethylsulfoxide. H Basch and H H Gadebusch Appl. Microbiol. 1968, 16(12):1953. These include: CONTENT ALERTS Receive: RSS Feeds, eTOCs, free email alerts (when new articles cite this article), more» Information about commercial reprint orders: http://journals.asm.org/site/misc/reprints.xhtml To subscribe to to another ASM Journal go to: http://journals.asm.org/site/subscriptions/ Downloaded from http://aem.asm.org/ on October 15, 2014 by guest Updated information and services can be found at: http://aem.asm.org/content/16/1 2/1953.citation APPuED MICRODJOLOGY, Dec. 1968, p. 1953 Copyright © 1968 American Society for Microbiology Vol. 16, No. 12 Printed In U.S.A. In Vitro Antimicrobial Activity of Dimethylsulfoxide H. BASCH AND H. H. GADEBUSCH Squibb Institute for Medical Research, New Brunswick, New Jersey 08903 Received for publication 1 October 1968 DMSO exerted a marked inhibitory effect on a wide range of bacteria and fungi, including one parasite, at concentrations likely to be encountered in antimicrobial testing programs in industry. The MMC generally was several-fold higher than the minimal inhibitory concentration, except for certain species which appeared to be ultrasensitive to this agent (Corynebacterium sp., Haemophilus influenzae, Pasteurella multocida, Herellea sp., M. tuberculosis var. BCG, Microsporum audouini). The pH of high concentrations of DMSO in water is likely to contribute to its innate microbicidal effect. On the basis of unpublished studies performed in our laboratories, it appears that basicity is a greater factor with aqueous mixtures of DMSO than with aqueous mixtures of acetone, the alcohols, or glycols. Microbiologically, DMSO compares favorably with the common solubilizing Downloaded from http://aem.asm.org/ on October 15, 2014 by guest Solubilization of drugs in dimethylsulfoxide (DMSO) before the determination of their antimicrobial activity is a common practice in the pharmaceutical industry. Since DMSO (Crown Zellerbach, Camas, Wash.) has been shown to possess bacteriostatic properties (2-4), a systematic documentation of its antimicrobial spectrum and level of activity was indicated. Conventional, serial twofold tube dilution tests were performed with double-strength Penassay Broth (Difco) for most bacteria and fungi; thioglycolate broth was used for Clostridia; Kirchner's medium (1) was used for Mycobacterium tuberculosis; Brain Heart Infusion broth (BBL) containing 2% rabbit blood was used for Diplococcus, Neisseria, Haemophilus, and Corynebacterium; S T S Medium (BBL) was used for Trichomonas foetus; and Desulfovibrio medium (Starkey) was used for Desulfovibrio. Most of the bacteria and yeasts, as well as the parasite, were agents. incubated for 24 hr at 37 C; Mycobacterium was LrmRAruRE Crrm incubated for 5 days; Desulfovibrio was incubated for 48 hr under nitrogen purge; and filamentous 1. Donovick, R., F. Pansy, G. Stryker, and J. Bernstein. 1950. The chemotherapy of experimental fungi were incubated for 48 hr at 26 C with metuberculosis. I. The in vitro activity of thiosemichanical agitation. carbazides, thiosemicarbazones, and related The microbicidal effect of DMSO was decompounds. J. Bacteriol. 59:667-674. termined by plating or by diluting out all tubes showing no growth by use of appropriate media. 2. Jacob, S. W., M. Bischel, and R. J. Herschler. 1964. Dimethyl sulfoxide (DMSO): a new conThe lowest drug concentration in which growth cept in pharmacotherapy. Current Therap. Res. failed to occur after subculture was considered to 6:134-135. be the minimal microbicidal concentration (MMC). Since aqueous solutions of DMSO were 3. Kligman, A. M. 1965. Topical pharmacology and toxicity of dimethyl sulfoxide (DMSO). J. Am. known to be alkaline, the pH was carefully moniMed. Assoc. 193:923-928. tored in each instance. Under the conditions of these experiments, the mean pH at the microbi- 4. Pottz, G. E., J. A. Rampey, and F. Benjamin. 1967. The effect of dimethyl sulfoxide (DMSO) on static concentration was 7.3 i 0.29 for bacteria and 7.1 i 0.39 for fungi. At the microbicidal antibiotic sensitivity of a group of medically imconcentration, the mean pH after incubation was portant microorganisms: preliminary report. 7.8 0.41 for bacteria and 7.7 0.41 for fungi. Ann. N.Y. Acad. Sci. 141:261-272. i 1953 1954 APPL. MICROBIOL. NOTES TABLE 1. Antimicrobial activity of dimethylsulfoxide Culture inoculum Organism No." MICc pHd MMCle 8 20 30 10 20 20 10 20 8 20 8 5 20 10 20 7 6 10 20 9 20 20 9 10 10 10 10 6 8 7.6 6.8 7.9 7.3 7.5 7.6 7.5 7.5 7.3 7.5 30 40 40 30 40 40 30 40 10 30 Pd Sizeb Bacteria 1,276 1,276 2,406 3,862 1,648 Staphylococcus aureus var. Geotrichum candidum Nocardia asteroides Cladosporium resinae Trichophyton mentagrophytes Penicillium notatum Fusarium bulbigenum Microsporum audouini Pullularia pullulans Parasite Trichomonas vaginalis 5,479 2,529 8,579 8,621 8,523 BCGrI 5,516 1,759 (H) (H) (H) (H) (H) 3,777 5,378 5,348 8,622 8,624 2,975 1,678 1,565 3,855 3,873 3,850 3,030 3,821 1,461 1,468 8,334 3,840 2,828 2,100 5,314 2,616 1,600 8,623 2,626 5,476 2,637 2,122 5,273 5,282 2,599 8,560 (H) (H) (H) (H) (H) (H) (H) (H) (H) (H) (H) (H) (H) 9 10 8 9 8 8 20 7 8 10 20 7 9 4 10 10 7.6 6.8 7.0 7.1 7.2 7.0 7.2 30 20 40 8 9 20 20 30 20 20 20 30 30 30 30 8 20 7.3 7.4 6.7 7.1 7.1 6.8 7.6 6.2 7.2 7.2 7.7 7.2 6.8 30 30 30 20 20 20 40 30 30 30 30 8 20 7.6 7.3 7.3 6.8 7.4 7.0 7.6 7.5 7.1 7.8 8.2 8.3 7.8 8.2 8.1 7.8 8.2 7.5 8.3 7.1 8.7 8.2 7.3 7.4 7.5 7.4 7.9 7.6 7.5 7.6 7.5 7.9 7.8 7.8 7.0 7.5 8.0 7.9 7.6 7.5 7.5 7.5 8.1 6.7 8.0 8.1 8.0 7.2 7.7 NTf a Squibb stock culture number. b (H) is equal to 106 cells/ml; for all other bacteria, Nocardia, and yeasts, the inoculum was 103 cells/ ml; for filamentous fungi, the inoculum was 2 ml of a 1:50 dilution of mat or spore suspension derived from a 48-hr growth (25 C) of the organism on 100 ml of Penassay agar in a 250-ml Erlenmeyer flask. c Minimal inhibitory concentration in DMSO (%). d After incubation. e Minimal microbicidal concentration in DMSO (%). f Not tested. Downloaded from http://aem.asm.org/ on October 15, 2014 by guest S. aureus S. aureus S. pyogenes S. faecalis Diplococcus pneumoniae Micrococcus lysodeikticus Sarcina lutea Corynebacterium sp. Listeria monocytogenes Mycobacterium tuberculosis Clostridium pasterianum Bacillus subtilis Desulfovibrio desulfuricans Neisseria catarrhalis Haemophilus influenzae Pasteurella multocida Escherichia coli Aerobacter aerogenes Klebsiella pneumoniae Proteus vulgaris P. mirabilis Salmonella schottmuelleri S. gallinarum S. typhimurium Shigella flexneri Serratia marcesens Herellea sp. Pseudomonas aeruginosa Fungi Aspergillus niger A. fumigatus Candida albicans C. krusei Saccharomyces cerevisiae