In vitro antimicrobial activity of dimethylsulfoxide.

In vitro antimicrobial
activity of
dimethylsulfoxide.
H Basch and H H Gadebusch
Appl. Microbiol. 1968,
16(12):1953.
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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On the basis of unpublished studies performed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
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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.
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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