Vol.

33

A Study of Lauryl Sulfate

Practical
Tryptose Broth for Detection of the
Presence of Coliform
Organisms in Water*
MAC H. McCRADY, F.A.P.H.A.
Chief of Laboratories, Quebec Ministry of Health, Montreal, Canada
THE studies, initiated by the Com- of certain non-coliform organisms.
mittee on Standard Methods in Cowles 6 first proposed the use of
1934, of media proposed for detection sodium lauryl sulfate for this purpose.
of the presence of coliform organisms in Mallmann and Darby7 employed it in
water, the results of which have been their preliminary work in the form of
reported at various times (1935,1. 2 Duponol W. A. Paste,* a commercial
193 7,3 and 1939 4), were continued dur- product used as a detergent and wetting
ing the past two years by conducting a agent in the textile and other industries.
practical test of another medium, lauryl A similar product, Duponol W. A.
sulfate tryptose broth, proposed by Flakes,t containing a somewhat larger
W. L. Mallmann and C. W. Darby.5'7 proportion of sodium lauryl sulfate,
was employed in the present study be-
THE MEDIUM cause of its apparently greater stability.
The composition of the Inedium
tested was the following: OUTLINE OF THE WORK
Grams The medium described was dis-
Bacto-tryptose 20 tributed, in dehydrated form, through
Bacto-lactose 5 the courtesy of Difco Laboratories,
Potassium phosphate dibasic 2.75 Detroit, Mich., to the following 17
Potassium phosphate monobasic 2.75
Sodium chloride state, provincial, county, city and uni-
Sodium Lauryl Sulfate (Duponol versity laboratories in the United States
WA Flakes) 0.1
The ingredients were dissolved in 1 liter *
Because of the excessive demands to which all
of cold distilled water, distributed into tubes laboratories were subjected following the outbreak of
and autoclaved at 15 lbs. pressure for 15 war, the labor involved in conducting this study and
minutes. The final pH was approximately 6.8. in compiling and reporting the results thereof, was
divided between Dr. W. L. Mallmann, Chairman of
the Standard Methods Committee on Water and
In this medium, tryptose was em- Sewage, and the writer. It was arranged that the
ployed, instead of peptone, because of former should initiate the study, enlist laboratories
to collaborate in the study and collect the results
its better productivity and selectivity; secured by them, whereupon these results were to
and sodium lauryl sulfate was in- be compiled and reported by the latter in his capacity
of Referee for the committee. Accordingly, the
cluded because of its selective effect, original results from the collaborating laboratories
were transmitted to the Referee early in the year, and
somewhat similar to that of some other from them was prepared the present report.
surface tension depressants such as t Duponol products are manufactured by E. I.
Du Pont de Nemours and Company, Wilmington,
oxgall, in inhibiting the multiplication Dela.
[11991
1200 AMERICAN JOURNAL OF PUBLIC HEALTH
and Canada, the directors of which
agreed to collaborate in the study:
Laboratories
Connecticut State
Maryland State
Minnesota State
Quebec Provincial
West Virginia State
Louisiana-Caddo Shreveport Health Unit
Indiana-Indianapolis Water Co.
Michigan-Detroit Filtration Plant
Highland Park "
Escanaba " S. J. Shank
Flint " "
Saginaw
Wyandotte
Missouri-St. Louis Public Health
California-Stanford University
Pennsylvania-Pennsylvania State College
University of Pennsylvania
The laboratory procedures specified
for the comparative work included,
briefly, planting of several portions
(usually five 10 ml. and one 1.0 ml.
portions) of sample in both lactose
broth and lauryl sulfate tryptose
broth; upon appearance of gas in these
primary media, eosin-methylene-blue
agar plates were streaked and colonies
appearing on the plates were fished to
both lactose broth and lauryl sulfate
tryptose broth, and also to agar slants
for microscopic examination. An addi-
tional transfer from gas positive tubes
of one or both primary media to bril-
liant green bile (2 per cent) was also
made by the majority of the collabo-
rating laboratories, and the formation
of gas in the bile, after incubation at
370 C. for 48 hours, was recorded.
It will be noticed that the procedures
specified included two departures from
the usual Standard Methods procedure:
the substitution of lauryl sulfate tryp-
tose broth for lactose broth as a primary
medium, and also its substitution as a
secondary medium in the "completed
test."
AGGREGATE RESULTS
Since some of the collaborating labora-
Oct.) 1943
tories made no transfers of primary
gas positive tubes to brilliant green bile,
Collaborators
Friend Lee Mickle
C.* A. Perry and A. A. Hajna
H. A. Whittaker and D. M. Taylor
Jacques Archambault
Katherine E. Cox
Alvin F. Meyer
C. K. Calvert and E. R. Hupp
W. M. Wallace; Fabien, and Rausch
Irving Dahljelm
C. H. Burdick and W. V. Kennedy
J. C. Richardson and A. C. Pendell, Jr.
Harry W. Ward
Jane I. Burns
Paul Beard
Michael A. Farrell
David Lackman
and some made such transfers only
from lactose broth, the results will be
reported under these headings: (A)
with no transfer from primary gas posi-
tives; (B) with transfer from lactose
broth primary gas positives; (C) with
transfer from both lactose broth and
lauryl sulfate tryptose broth gas posi-
tives. Consequently (B) will include
(C), omitting from the latter the re-
sults of transfer to brilliant green bile
from lauryl sulfate tryptose primary
gas positives; and, likewise, (A) will
include both (B) and (C), omitting
from both of the latter the results of
transfer to brilliant green bile.
Table 1 shows the aggregate results,
reported by the various collaborating
laboratories, from samples that yielded
one or more primary gas positives. A
large number of other samples were ex-
amined but, as they gave no gas posi-
tives when planted in the two primary
media, are not included in the tabula-
tions.
Table 1-A-A total of 17 labora-
tories reported the results shown in
Table 1-A. It will be observed that,
from 605 water samples yielding one
or more primary gas positives, 1,873 of
3,415 portions of sample, or 54.8 per
Vol. 33 LAURYL SULFATE TRYPTOSE BROTH 120)1

TABLE 1
Gas Positives Completely Gas Positive
Confirmed Using Secondary upon Transfer
Portions of Primary to
Gas Positive Lactose Broth L.S.T. Broth Brilliant Green Bile
No. of /C- w
Sample Per cent of Per cent of Per cent of Per cent of
Primary Portions Sample Sample Sample Sample
Medium Planted No. Portions No. Portions No. Portions No. Portions
A. Aggregate Results of Examination of 605 Water Samples Yielding Primary Gas Positives
With No Transfer of Primary Positives to Brilliant Green Bile
Lactose broth 3,415 1,873 54.8 1,117 32.7 1,142(1) 33.4
L.S.T. broth 3,415 1,628 47.7 1,222(3) 35.8 1,261 36.9
B. Aggregate Results from 526 Water Samples Yielding Primary Gas Positives With Transfer
of Lactose Broth Primary Positives to Brilliant Green Bile
Lactose broth 2,996 1,687 56.3 1,025 34.2 1,052(1) 35.1 1,088(2) 36.3
L.S.T. broth 2,996 1,486 49.6 1,115(3) 37.2 1,150 38.4
C. Aggregate Results from 318 Water Samples Yielding Primary Gas Positives With Transfer
of Lactose Broth and Lauryl Sulfate Tryptose Broth Primary Positives to Brilliant Green Bile
Lactose broth 1,732 1,016 58.7 540 31.2 527(1) 30.4 579(2) 33.4
L.S.T. broth 1,732 775 44.7 586(3) 33.8 575 33.2 601(1 34.7
Note: The figures in parentheses (which are included in the adjoining totals) represent the nuinber of missing
tubes, incomplete results, etc., which were considered positive in order to avoid loss of the remaining
data obtained from these particular samples; a study of these data suggests that the error thus intro-
duced is very probably negligible.

cent, planted in lactose broth, gave gas The number of complete confirma-
positives; whereas, 1,628 of a like num- tions of coliform organisms (1,117) ob-
ber of sample portions, or 47.7 per cent, tained by the use of the usual pro-
planted in L.S.T. (lauryl sulfate tryp- cedure, employing L.B. (lactose broth),
tose) broth yielded gas positives. When compares with the numbers secured by
these primary positives were plated on the use of L.S.T. broth at various stages
eosin-methylene-blue agar and colonies of the procedure as follows:
fished to lactose broth and to L.S.T. Primary Secondary No. Per cent
broth, and to agar slant for microscopi- L.B. L.B. 1,117 100.0
cal examination, the 1,873 lactose broth L.B. - L.S.T. 1,142 102.2
primary positives yielded 1,117 com- L.S.T. - L.B. 1,222 109.4
plete confirmations of coliform organ- L.S.T. - L.S.T. 1,261 112.9
isms using secondary lactose broth,
and 1,142 complete confirmations using Thus the use of L.S.T. broth, as
lauryl sulfate tryptose broth. Conse- primary and as secondary in the com-
quently, the substitution of L.S.T. pleted test, permitted an increase of
broth for lactose broth as a secondary 12.9 per cent in the number of complete
medium yielded 25, or slightly over 2 confirmations compared with the usual
per cent, more complete confirmations. procedure using lactose broth in these
The use of lauryl sulfate tryptose positions, despite a reduction of 13.1
broth as a primary medium reduced the per cent in the number of primary gas
number of primary gas positives from positives that had to be examined when
1,873 to 1,628, or slightly over 13 per using L.S.T. broth.
cent, but yielded 1,222 complete con- It is evident that this series of re-
firmations when using lactose broth as sults indicates an increase in the num-
a secondary medium, and 1,261 when ber of sample portions containing coli-
using L.S.T. broth as a secondary forms detected through the use of
medium. L.S.T. broth, both as a primary and as
1202 AMERICAN JOURNAL OF PUBLIC HEALTH
a secondary medium, which, although
not very great, is nevertheless signifi-
cant.
Lauryl sulfate tryptose broth was
suggested as a primary medium princi-
pally because it gave promise of serving
as a presumptive medium in which the
formation of gas might be considered
a practically sure indication of the
presence of coliform organisms. In
this respect, however, only 77.5 per
cent of the L.S.T. primary gas positives
yielded coliforms, even when using
L.S.T. broth as a secondary medium,
compared with 61.0 per cent of com-
plete confirmations secured from lactose
broth primary gas positives using the
same secondary medium, and 59.6 per
cent using lactose broth as the second-
ary medium. These relationships are
shown below in tabular form:
Complete
Primary Confirmations
Pri- Second- Gas
mary ary Positives No. Per cent
L.B. -L.B. 1,873 1,117 59.6
L.B. -L.S.T. 1,873 1,142 61.0
L.S.T.-L.B. 1,628 1,222 75.1
L.S.T.-L.S.T. 1,628 1,261 77.5
It is interesting to compare these
figures with those obtained in a previous
study,3 reported in 1937, of the utility
of fuchsin broth as a primary medium,
with lactose broth the secondary me-
dium, when a number of laboratories
collaborated in the examination of 890
waters from various sources. Of 2,903
primary gas positives in fuchsin broth,
70.5 per cent were completely con-
firmed, whereas of 4,204 lactose broth
primary positives, 50.2 per cent were
completely confirmed. The number of
complete confirmations from fuchsin
broth primary gas positives was 97.06
per cent of that from lactose broth
primary gas positives, whereas the pres-
ent study indicates the number of com-
plete confirmations, using L.S.T. broth
as primary and secondary media, was
112.9 per cent of that from lactose
Oct., 1943
primary gas positives. Obviously, to
judge from these figures, L.S.T. broth
is the better medium of the two; but,
as in the use of fuchsin broth, primary
gas positives in L.S.T. broth must also
be confirmed.
It has been suggested 5 that the
plating medium, eosin-methylene-blue
agar employed in the Standard Methods
procedure for the "completed test,"
inhibits the growth of some coliform
organisms; and that, for this reason,
gas in primary fermentation tubes con-
stitutes evidence of the presence of
coliform organisms more frequently
than successful complete confirmation
indicates. This contention is indeed
probable, but the magnitude of the error
thus introduced is problematical. A
previous study 2 showed, by transfer
from primary gas positives to various
liquid media followed by complete con-
firmation of gas positives in the latter,
that the error of the usual completed
test performed on primary lactose broth
positives averaged, at least, about 14
per cent, even when employing eosin-
methylene-blue agar in the complete
confirmation of the liquid confirmatory
gas positives. If a practicable plating
medium less inhibitive than E.M.B.
agar can be found for use in the "com-
pleted test," the present procedure of
complete confirmation may have to be
debited with an additional error; but
until the magnitude of the error due to
the use of E.M.B. agar is definitively
determined, there appears to be no
alternative to acceptance of this plating
medium in the "completed test."
Finally, the aggregate results shown
in Table 1-A indicate that L.S.T. broth
could not be employed as a presumptive
test of the samples examined without
confirmation, but that its use both as
primary and secondary medium in the
"completed test" permitted about 13
per cent more complete confirmations
than did the use of lactose broth, with
a reduction of about 13 per cent in the
V/ol. 33 LAURYL SULFATE TRYPTOSE BROTH
number of primary gas positives that
had to be confirmed.
Table 1-B-Fifteen laboratories in-
cluded a transfer from primary lactose
broth gas positives to brilliant green
bile in their examination of 526 water
samples. It will be observed that the
relationships between the " completed
test " results, shown in Table 1-B, se-
cured from the use of lactose broth and
lauryl sulfate tryptose broth in the
examination of these samples, resemble
fairly closely those described in the dis-
cussion of the results of Table 1-A.
Use of the " confirmatory test," em-
ploying transfer to brilliant green bile
from lactose broth primary gas positives
and considering formation of gas in the
bile to constitute evidence of the presence
of coliform organisms, indicated 36.3
per cent of the sample portions planted
to contain these organisms. This pro-
portion is slightly higher than the 34.2
per cent of complete confirmations
secured by the usual Standard Methods
procedure, and the 35.1 per cent by the
same procedure using L.S.T. broth in-
stead of lactose broth as the secondary
medium. It is slightly lower, however,
than the proportion of 37.2 per cent
obtained from the use of L.S.T. broth
as the primary medium, and that of
38.4 per cent secured by employing
L.S.T. broth both as primary and sec-
ondary media.
Table 1-C-Eleven laboratories in-
cluded transfers from both lactose broth
and L.S.T. broth primary gas positives
to brilliant green bile in their examina-
tion of 318 water samples.
The results of this series, given in
Table 1-C, show that the proportions
of water sample portions in which the
presence of coliform organisms was in-
dicated by these two "confirmatory"
procedures compared with the complete
confirmation procedures as shown
above, right.
Evidently the indications of the pres-
ence of coliform organisms furnished by
1203
Per cent of
Gas Water Sample
P, i- Second- in Portions
mary dary B.G.B. Positive
L.B. -L.B. 31.2
L.B. -L.S.T. 30.4
L.S.T.-L.B. 33.8
L.S.T.-L.S.T. 33.2
L.B. + 33.4
L.S.T. + 34.7
gas formation in brilliant green bile, fol-
lowing transfer from either lactose broth
or L.S.T. broth primary gas positives,
were slightly less numerous (33.4 per
cent) and slightly more numerous (34.7
per cent), respectively, than those
(33.8 per cent) furnished by the com-
plete confirmation of L.S.T. primary
positives using lactose broth secondary,
which in this series of examinations
yielded the best of the "completed
test" results.
It may be concluded, therefore, from
the aggregate results of Table 1, that
L.S.T. broth could not be employed in
a presumptive test, without confirma-
tion, of these waters; that the substitu-
tion of L.S.T. broth both as primary
and secondary media yielded somewhat
more frequent complete confirmation of
the presence of coliform organisms
than the other complete confirmation
procedures, and about the same number
of indications of the presence of coli-
forms as that from the use of lactose
broth primary followed by the brilliant
green bile confirmatory test (gas only);
and that the use of L.S.T. primary fol-
lowed by the brilliant green bile con-
firmatory test furnished, by a small
margin, the greatest number of indica-
tions of the presence of coliforms.
RESULTS FROM DIFFERENT TYPES
OF WATERS
The data summarized in Table 1 are
distributed in Table 2 according to type
of water examined. Under each type
heading are given three sets of figures
corresponding to the A, B, and C
1204 AMERICAN JOURNAL OF PUBLIC HEALTH Oct., 1943

groups of Table 1, the first set showing
the results secured when no transfer of
primary gas positive to brilliant green
bile was made, the second set when such
transfer was made from lactose broth
primary gas positives, the third set
when such transfer was made from
both primary media gas positives. The
first set of figures includes the second
and third, and the second set includes
the third, as in Table 1.
It will be observed in Table 2 that
the results secured from the various
types of waters present as a rule the
same general picture as that furnished
by the aggregate results. The L.S.T.

TABLE 2
Results of Examination of Different Types of Waters
Complete Confirmations Gas in
Using Secondary B.G. Bile
Portions Gas t -
-- A upon Transfer
Primary No. of Sample Positive L.B. L.S.T. from Primary
Raw Medium Portions Planted Per cent Per cent Per cent Per cent
12 laboratories L.B. 750 66.9 46.5 45.7
139 samples L.S.T. 750 64.1 48.8 48.8
12 laboratories L.B. 695 67.2 46.8 45.8 46.8
128 samples L.S.T. 695 64.0 48.1 48.1
9 laboratories L.B. 515 71.3 47.2 45.6 48.4
98 samples L.S.T. 515 65.2 48.2 46.8 47.6
Unfinished
10 laboratories L.B. 535 50.7 15.3 15.3
99 samples L.S.T. 535 21.1 15.9 16.3
10 laboratories L.B. 517 50.1 15.5 15.5 17.4
95 samples L.S.T. 517 21.7 16.2 16.6
8 laboratories L.B. 481 53.4 16.6 16.6 18.7
92 samples L.S.T. 481 21.8 17.5 17.7 18.9
Finished
9 laboratories L.B. 434 37.6 10.8 11.1
77 samples L.S.T. 434 16.1 11.8 11.5
8 laboratories L.B. 410 39.3 11.5 11.7 12.0
73 samples L.S.T. 410 16.3 11.7 11.5
6 laboratories L.B. 332 41.3 -. 10.2 10.8 11.1
60 samples L,S.T. 332 16.9 12.3 11.4 13.9
Pools
S laboratories L.B. 310 35.8 17.7 16.8
58 samples L.S.T. 310 24.5 19.4 20.0
3 laboratories LB. 90 38.9 31.9 31.9 30.0
15 samples L.S.T. 90 37.8 34.4 34.4
3 laboratories L.B. 78 43.6 34.6 34.6 33 .3
13 samples L.S.T. 78 41.0 38.5 38.5 38.5
Wells Springs
7 laboratories L.B. 1,195 61.0 44.5 45.8
200 samples L.S.T. 1,195 66.0 50.7 52 .3
6 laboratories L.B. 1,099 60.9 44.9 46.2 48.0
184 samples L.S.T. 1,099 66.3 51.3 52.9
3 laboratories L.B. 308 69.1 48.7 46.8 55.8
52 samples L.S.T. 308 76.6 56.8 56.2 58.8
Miscellaneous
(type not reported)
6 laboratories LT. 191 50. 8 27.2 36.6
3 2 samples L.S.Tr. 191 51.8 28.3 37 .2
5 laboratories L.I. 185 51.9 28.1 3 7. C 37.8
3 1 samples L.S.T. 185 53.5 29.2 3S .4
2 laboratories L.B. 19 44.4 4.44. 27.8 27.8
3 samples L.S.T. 18 55.6 44.4 44.4 44.4
Sea Water
1 laboratory L.B. 40 45.O 40.0 42.5 45.0
10 samples L.s.r. 40 55.0 50.0 so. 0 ....
Vol. 33 LAURYL SULFATE TRYPTOSE BROTH 1205

broth primary gas positives were usually wells and springs, and other waters,
less numerous than the lactose broth however, this substitution increased the
gas positives, 56 to 58 per cent less number of positive complete confirma-
numerous in the case of unfinished and tions by 13 to 37 per cent.
finished waters, although slightly more The "confirmatory tests," employing
numerous in the case of well and spring, transfer from primary gas positives to
miscellaneous, and sea waters. brilliant green bile, indicated as a rule
The proportion of L.S.T. primary gas about the same frequency of presence
positives that were shown by the "com- of coliform organisms as did the best
pleted test" (using L.S.T. broth as of the "completed tests," with one ex-
secondary medium) to contain coliform ception: four (Minnesota, Pennsylvania
organisms, varied from about 70 per State College, Quebec, West Virginia)
cent for finished waters to 82 per cent of the six laboratories performing "con-
for swimming pool waters, the percent- firmatory tests" of well and spring
age usually ranging between 75 and 80. waters reported a total of positive "com-
Evidently the formation of gas in pleted tests," using L.S.T. broth
primary L.S.T. broth could not be con- primary and secondary, which was
sidered very sure proof of the presence about 16 per cent greater than that of
of coliform organisms. "confirmatory tests" with brilliant
As a rule there was very little differ- green bile following lactose broth
ence between the results of employing primary. An interesting feature of the
lactose broth and L.S.T. broth as reports from these four laboratories on
secondary media in the "completed ground waters was the greater number
test." of primary gas positives in L.S.T. broth
The use of L.S.T. broth as both (554) compared with that in lactose
primary and secondary media in the broth (509). Generally, however, the
"completed test," however, permitted greatest number of positive indications
isolations of coliforms from more sample of the presence of coliforms was secured
portions than did the use of lactose by the use of L.S.T. broth primary fol-
broth, as indicated below: lowed by "confirmatory" brilliant green
bile, taking formation of gas in the
Per cent of Positive to constitute a positive result.
Completed Tests Using latter
Usual Standard Method Consequently, the results from the
Type of Water (Lactose Broth) various types of water samples ex-
Raw 105 amined confirm in general the conclu-
Unfinished 106 sions derived from the aggregate re-
Finished 106 sults. Furthermore, the separate groups
Swimming, pools 113 of results reported by the various col-
Wells and springs 117
Sea water 125 laborating laboratories, with few ex-
Miscellaneous 137 ceptions, also confirm these conclusions.
These figures do not indicate much FINISHED WATERS
advantage, as regards positive complete Since the use of L.S.T. broth appears
confirmation, in substitution of L.S.T. to reduce the number of primary gas
broth for lactose broth in the examina- positives that must be confirmed, par-
tion of waters from purification plants, ticular interest attends its employment
but it must be remembered that fewer in the examination of finished waters,
primary positives had to be confirmed which usually yield an excessive pro-
when L.S.T. broth was employed. In portion of false lactose broth primary
the examination of swimming pools, gas positives. Consequently Table 3
1206 AMERICAN JOURNAL OF PUBLIC HEALTH Oct., 1943

TABLE 3
Results of Examination of Finished Waters
Complete Confirmations Gas in
Sample Primary Using Secondary B.G. Bile
Primary Portions Gas r -- upon Transfer
Laboratory Medium Planted Positives L.B. L.S.T. from Primary
Indianapolis, Ind. L.B. 97 56 0 0 0
L.S.T. 97 0 0 0 0

Maryland L.B. 42 16 6 6 6
L.S.T. 42 13 13 11 13
Penn. State Coll. L.B. 24 12 3 4 5
L.S.T. 24 10 6 5 8
Penn. University L.B. 103 41 25 26 26
L.S.T. 103 32 22 22 25
Saginaw L.B. 30 6 0 0 0
L.S.T. 30 0 0 0 0

Wyandotte, Mich. L.B. 36 6 0 0 0

L.S.T. 36 1 0 0 0

Minnesota L.B. 42 12 8 8 8

L.S.T. 42 8 5 7
Quebec L.B. 30 11 5 4 4
L.S.T. 30 2 1
Louisiana L.B. 12 1 0 0
L.S.T. 12 3 3 3

is presented, showing the results re-
ported by those laboratories that ex-
amined at least a few finished water
samples.
Of the six laboratories, listed in
Table 3, that reported coliforms in
finished waters, three isolated more by
the use of the usual Standard Method
procedure using lactose -broth than by
the use of L.S.T. broth primary,
whereas the experience of the other
three laboratories was the reverse.
These data are too few in number to
permit a conclusion regarding the ap-
plicability of lauryl sulfate tryptose
broth to the examination of finished
waters. Much more information is
needed to establish whether the medium
inhibits coliform organisms that have
been subjected to purification processes.
CONCLUSIONS
Seventeen laboratories situated in
the United States and Canada have
collaborated in a study of the utility of
lauryl sulfate tryptose broth for de-
tection of the presence of coliform or-
ganisms in water. A total of 605 water
samples from which primary gas posi-
tives were obtained, was examined, em-
ploying L.S.T. broth as a primary and
as a secondary medium for comparison
with lactose broth in the Standard
Methods "completed test" procedure.
Of these samples, 526 were also sub-
jected to the "confirmed test" procedure
employing transfer from lactose broth
primary gas positives to brilliant green
bile; and 318 were examined further
by transfer from L.S.T. broth primary
gas positives to brilliant green bile.
The results reported by the col-
laborating laboratories indicated that,
in general:
1. A reduction of about 13 per cent in the
number of sample portions producing gas
in the primary medium resulted when L.S.T.
broth was substituted for lactose broth, this
reduction being particularly evident in the
results from unfinished, finished and swim-
ming pool waters.
2. As coliform organisms were isolated from
only 75.1 per cent of L.S.T. broth primary
gas positives when using lactose broth second-
ary, and from 77.5 per cent when using
L.S.T. broth secondary in the usual Standard
Methods " completed test" procedure, it is
evident that L.S.T. broth could not be em-
ployed as a presumptive medium, without
confirmation, in the examination of these
waters.
3. Since the use of L.S.T. broth as both
primary and secondary media permitted iso-
Vol. 33 LAURYL SULFATE TRYPTOSE BROTH
lation of coliform organisms from 1,261 sam-
ple portions, about 13 per cent more than
the 1,171 portions from which isolations were
secured when lactose broth was employed,
the substitution of the former medium for
the latter appeared to be advantageous; but
the paucity of data reported on finished
waters precludes a definitive conclusion at this
time regarding the utility of L.S.T. broth in
the examination of this important group of
waters.
4. The use of the " confirmed test " (trans-
fer of primary gas positive to brilliant green
bile, taking gas in the latter to indicate the
presence of coliform organisms) yielded about
as many positive results as did any of the
"completed tests," whether transfer was made
from the lactose broth or the L.S.T. broth
primary gas positives. Transfer from the
L.S.T. broth, however, resulted in a small
increase (3.8 per cent) in the number of posi-
tive " confirmed tests " compared with trans-
fer from lactose broth gas positives.
On the whole, the substitution of
lauryl sulfate tryptose broth for lac-
tose broth in the Standard Methods
procedures promises a reduction in the
number of primary gas positives to be
1207
confirmed and an increase in the num-
ber of positive indications of the pres-
ence of coliform organisms. Conse-
quently further study of the medium by
interested laboratories is recommended,
particularly with reference to its util-
ity in the examination of finished
waters.
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in Water Analysis. J. Am. W. W. Assoc., 27,
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3. McCrady, M. H. A Practical Study of Procedures
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