766 NOTES [VOL.

M5
In order to test this, Mycobacterium,lacicola was grown in a medium already
described. Forty mg per cent neutralized benzoic acid was added to the com-
plete medium. After 48 hours' growth at room temperature the bacteria were
centrifuged and washed, and a standard suspension was made. Aliquots were
placed in Warburg vessels, and the oxygen uptake was measured in m/20 phos-
phate buffer pH 6.7 in the presence of benzoic acid with and without different
amounts of streptomycin. Table 1 shows that the oxidation of benzoic acid
by organisms grown in the normal medium is inhibited 44 per cent by 50 pAg per
ml, 82 per cent by 100 jug per ml, and completely by 200 pug per ml of strepto-
TABLE 1
The effect of streptomycin on the oxidation of 1.0 mg sodium benzoate by M. lacticola at 37 C
pH 6.7
TIME NORMAL GROWN IN 40 MG % BENZOATS

Min Control 50 Ag S 100lg S 200pug S Control 50 Ag S 100 g S 00gS

30 -6 -10 -4 -6 93 71 64 66
60 4 -2 -4 -8 276 216 195 185
90 59 28 9 -9 427 332 301 279
120 139 73 25 -6 554 449 408 379
150 229 119 40 -7 563 526 478 441
The oxygen uptakes in the absence of benzoate have been subtracted. The figures are
in terms of mms of oxygen utilized. S = streptomycin. The minus sign indicates an
inhibition of the resting respiration by the benzoate.

mycin. On the other hand, the oxidation of benzoic acid by organisms grown
in its presence is inhibited 25 per cent by 50 pug per ml, 30 per cent by 100 p&g
per ml, and 33 per cent by 200 pg per ml. Similar results are obtained when
p-hydroxybenzoic acid is used. These experiments suggest that the enzymes
are relatively insensitive to streptomycin and that inhibition is caused by the
suppression of their formation. The previously published experiment that
showed that streptomycin caused less inhibition in the presence of 2.0 mg of
benzoic acid than in the presence of 1.0 mg could be interpreted to mean that
2.0 mg caused the production of more enzyme.

STEROID EFFECT UPON BACTERIAL GROWTH

ESTHER W. SQUIRE AND EDWARD N. SQUIRE
Department of Biology, Mundelein College, Chicago, Illinois
Received for publication February 26, 1948
Raab (Science, 103, 670) recently reported that cholesterol added to culture
media inhibited the growth of Staphylococcus aureus. In this laboratory, 0.1 per
cent cholesterol in nutrient broth either inhibited or enhanced the growth of cer-
19481 NOTES 767
tain gram-negative organisms but was without effect upon the gram-positive
bacteria tested.
TABLE 1
Growth of bacteria in nutrient broth containing steroids*
CHOLES-
BICHIO- CEOLES- CEOLES- CHOLES- TERTh 5-cEo- 3,50-co
TERYL
LESTERYL CHLORDE TERYL
TEROL p-TOLU- LESTENE LESTA- CONTROL.
BROMIE ENE SUL- DIENE
PONATE

Alcaligenes faecalis........ N N N E N N N N
Staphylococcus aureus...... N N E N N N N N
Aerobacter aerogenes....... N E N I N N E N
Eacherichia coli............ N E E N N N I N
Prot&us vulgaris............ N N N N N N N N
Eberthella typhosa.......... N E N N N N N N
Shigella paradysenteriae.... N E N I N N N N
Bacillus subtilis............ N N N N N N E N
Salmonella schottmuelleri... N E E E N N N N
Staphylococcus albus........ N N N N N N E N
Klebsiella pneumoniae...... N I I I N N E N
Streptococcus lactis......... N I I N I I N N
Bacillus megatherium....... N N N N N N N N
Serratia marcescens......... N N N N N N N N
Sarcina lutea. N N N N N N N N
N - normal growth. E - enhanced growth. I - inhibited growth.
* Nutrient broth contained 0.1 per cent steroid derivative. Readings were measured by
turbidity.
The steroid derivatives, which were prepared by replacing the hydroxy group
of cholesterol with halogen or other substituents, generally affect the growth of
bacteria differently. This is shown in table 1.
The action of cholesteryl chloride and bromide is particularly interesting in
view of the fact that Turfitt (J. Bact., 54, 557) found many soil organisms unable
to utilize cholesteryl chloride.