Heat resistance of

Salmonella
typhimurium and
Salmonella
senftenberg 775W in
milk chocolate.

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J M Goepfert and R A Biggie

Appl. Microbiol. 1968,
16(12):1939.

Vol. 16, No. 12

Printed in U.S.A.

APPUED MICROBIOLOGY, Dec. 1968, p. 1939-1940

Copyright 1968 American Society for Microbiology

Heat Resistance of Salmonella typhimurium and

Salmonella senftenberg 775W in Milk Chocolate,
J. M. GOEPFERT AND R. A. BIGGIE
Food Research Institute, and Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706

1939

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Received for publication 4 October 1968

The problem of Salmonella contamination in temperature in Fig. 1. The ZD values of 32.4 and
dried foods has underscored our lack of knowl- 34.2 are almost twice those generally obtained for
edge concerning the heat resistance of enteric spores exposed to moist heat. Ng (1) determined
microorganisms in dry and semidry environments. that the moist heat resistance (D values) of S.
Confectionery manufacturing is one of the food senftenburg 775W and S. typhimurium was 31 min
industries concerned with dry-heat destruction of
Salmonella. The discovery of these organisms in
3
several chocolate-containing products in the
recent past has raised the question of whether
S TYPHIMURIUM
they can be killed by heating the molten chocolate
prior to casting. The results of experiments perZD= 34.2
formed to determine the heat resistance of Salmonella in miLk and dark chocolate are presented in
this paper.
S. typhimurium and S. senftenberg 775W were z
inoculated into milk chocolate in the following
manner. The cells resulting from 24-hr growth in
600 ml of Trypticase Soy Broth (BBL) incubated z
statically at 37 C were harvested centrifugally, w
resuspended in 20 ml of sterile skimmed milk, and
12lyophilized. Commercially formulated milk
chocolate was dispensed in Sorvall Omnimixer
S. SENFTENBERG 775
cups in 100-g quantities and warmed to the test
o
~~ZD= 32.4
temperature in an oil bath. The lyophilized cells
were added to the molten chocolate and blended LI
for 1 min at top speed on the Omnimixer. The
cups were returned to the oil bath and immersed
so that the level of the chocolate was /4 (1.9 cm)
inch below the surface of the oil. Agitation of the
inoculated chocolate throughout the remainder
of the experiment was accomplished by a mechanical stirrer.
Periodically, 1-ml samples were taken by using
wide-bore pipettes and a vacuum-pressure pump.
Dilutions were made in sterile 30% sucrose, and
viable salmonellae were enumerated by the threetube most probable number technique. Nutrient
FIG. 1. Plot of log D value vs. temperature for S.
broth was used as the recovery medium, and senftenberg
775W and S. typhimurium heated in milk
tubes showing turbidity after 48 hr at 37 C were chocolate. Points represent averages of the times given
streaked on MacConkey Agar (Difco) and incu- in Table 1.
bated at 37 C. Isolates were confirmed biochemically and serologically.
and 1.2 min, respectively, at 57 C. Although a
The D values obtained in replicate trials at direct
comparison of D values at 57 C is not
70, 80, and 90 C are presented in Table 1. The possible,
can be observed (Table 1) that an
averages of the D values are plotted against enormousitincrease
in heat resistance occurs when
I Publshed with the approval of the Director of the
salmonellae are exposed to dry heat. These data
Wisconsin Agricultural Experiment Station.
stress the inadvisability of extrapolating heat-

1940

NOTES

TABLE 1. D values of S. senftenberg 775W and S.

typhimurium (the results of three trials
at each temperature)
D values (min)
Temp (C)

S. sewfelberg S. typhimurium
775W

360
480
480

720
678
1050

80

144
96
108

222
222
222

90

36
30
42

78
72

The curves obtained in this study characteristically showed a rapid decline in numbers of
survivors (3-log cycles) during the first few
minutes of heating, followed by a slower rate of
decrease thereafter. The rapid initial loss might be
attributed to the death of cells injured during the
lyophilization and inoculation processes. This
problem was obviated by taking the first sample
at least 30 min after the addition of the cells to the
chocolate. Occasionally, curves were obtained
that showed a "tail," but attempts to isolate more
resistant organisms from this portion of the curve
were unsuccessful. More often than not, a
straight-line (logarithmic) relationship was obtained over the entire range of the test after the
initial delay in sampling.
The heat resistance of Salmonella was essentially the same in either milk or dark chocolate.
The degree of heating required to kill Salmonella in dry environments should lead to a reassessment of the heat treatments given to many
food products during manufacture. It now seems
possible that Salmonella may survive processing
treatments that formerly were assumed to be
adequate to ensure their destruction.

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70

APPL. MICROBIOL.

resistance values obtained under one set of conditions to any other conditions.
S. typhimurium was more resistant to dry
heating than S. senftenberg 775W. Ng (1) reported
that the latter serotype was approximately 25
times as resistant to moist heat as S. typhimurium,
a value confirmed in our laboratory. Since we
This investigation was supported by the National
examined only two serotypes, it is difficult to
predict if strains will be uncovered that are more Confectioners Association, Chicago, Ill.
resistant than S. typhimurium. These findings
LrrERATuRE Crrm
warn against establishing time-temperature relationships for dry-heat processes based on destruc- 1. Ng, H. 1966. The destruction of salmonellae. Agrition of S. senftenberg 775W.
cultural Research Service 74-37, p. 39-41.