Microbioogical Process Discussion
Calculation of Heat Sterilization Times for Fermentation Media
FRED H. DEINDOERFER1
Merck Sharp & Dohme Research Laboratories, Merck & Co., Inc., Rahway,
Received for publication September 14, 1956
Sterilization of nutrient media is an operation
essential to all industrial fermentation processes re-
quiring pure culture maintenance. AMethods for steri-
lizing nutrient media include 1) heating, 2) filtration,
3) irradiations, 4) sonic vibration, and 5) exposure to
chemical agents. Because heating the medium, usually
by steam, is the most reliable and on a large scale the
easiest to control, it is the method of choice throughout
the fermentation industry. The increasing availability
of radioactive isotopes undoubtedly will stimulate
further study of the use of gamma radiation for sterili-
zation. However, a great deal of work is needed before
commerically feasible gamma ray applications will be
used in the fermentation industry. The other methods,
although successful on a laboratory scale, present too
many operational drawbacks for large-scale use in
fermentation processes.
How long should a nutrient medium be exposed at
high temperatures to achieve sterile conditions? This
question arises often and usually is answered in bench-
scale or pilot plant tests. Improper translation of these
test results with large equipment may subject the
medium to unnecessary overheating. A method by
which minimum exposure time to achieve sterile condi-
tions can be calculated from easily obtainable thermal-
death relationships and the temperature conditions in a
heat sterilization process is presented in this paper. The
method can be used to correlate sterilization conditions
among various sized fermentation vessels. It also per-
mits evaluation of the temperature and retention time
relationship in continuous sterilizers. It need not be
restricted to fermentation processes, but should be
applicable to any process involving a heat sterilization
or pasteurization operation.
Thermal Resistance of Microorganisms
The ability of microorganisms to withstand heat is
much greater than that of other forms of life. Thermo-
philes capable of even tolerating common heat sterili-
zation conditions, such as exposure to steam at 250 F
for 20 to 30 min, exist. Their occurrence in nutrient
media, however, is rare. Relative resistances of several
microorganisms to moist heat are shown in table 1.
1 Present Address: E. R. Squibb & Sons, New Brunswick,
New Jersey.
221
New Jersey
Bacterial spores are by far the most heat resistant forms
of microorganisms.
The thermal resistance of bacterial spores is in-
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herently different among species and even among strains
of the same species. Besides thermal resistance inherent
to spores of a particular species, variations can be caused
by a number of environmental factors. These can be
generalized into two groups: 1) environmental factors
affecting sporulation prior to sterilization, and 2)
environmental factors affecting spores during the
sterilizing heat exposure. The most important factors
in nutrient media sterilization fall in the latter category
and include factors such as the pH during sterilization
and the osmotic nature of the media. The presence of
suspended solids also affects sterilization by physically
insulating spores from heat exposure.
Thermal-Death Relationships of Bacterial Spores
Because bacterial spores are the most heat resistant
forms of microorganisms, their germinated cells are the
most frequent contaminants encountered in industrial
fermentations due to improper sterilization. The ensuing
discussion will deal, therefore, with the thermal-death
relationships of these forms. Relationships between the
number of viable spores and exposure time to heat
demonstrate a logarithmic rate of spore viability de-
struction. Two typical survivor curves for spores of
Bacillus stearothermophilus strain 1518, an organism
often used for sterilization studies in the food industry,
are shown in figures 1A and lB. These curves were
obtained in buffer solutions and would differ from their
respective curves in other media. Spore destruction at
the higher temperature is more than 400 times faster
than at the lower temperature.
TABLE 1. Relative resistances of microorganisms to sterilization
by moist heat*
Organism Organism ~~~~~~Relative
Resistance
Escherichia coli ............................. 1
Bacterial spores ............................. 3,000,000
Mold spores ................................. 2-10
Viruses an(d bacteriophages ................. 1-5
*
Rahn (1945).
222
The curves in figures IA and lB represent first-order
reactions, where the rate of spore destruction is directly
proportional to the number of surviving spores. This
can be described mathematically as follows:
dN
dO (1)
where N is the number of surviving spores in the volume
under consideration, 0 is the time of exposure, k is a
velocity constant, and dNdO
is the rate of spore destruc-
tion.
By integrating this equation, it can be modified to a
form that can be used to calculate the time for steriliza-
F. H. DEINDOERFER [VOL. 5
spores of a particular species will be a function of
temperature only. It will behave similarly to other velo-
city constants in its relation with temperature, quantita-
tively expressed by the empirical Arrhenius equation
as follows:
k = Ae RT (3)
where A is a proportionality constant, R is the gas
constant, T is the absolute temperature, and ,u is an
apparent activation energy for heat destruction of the
spores. No theoretical significance need be attached to
this equation for this discussion. The conformity of
many destruction rates to the Arrhenius equation

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tion at a particular temperature where the velocity
constant, k, is known. Then,
6=2.3l
k N1
N (2)
where N1 is the number of particular spores at the start
of the heat exposure and N2 is the number of the same
spores surviving at any time, 0, after exposure has
started. N2 in this relationship can never reach zero,
but practikally this does not matter. For destruction of
spores wAith only one chance in one hundred of failing to
destroy all spores, N2 should be set equal to 0.01.
Higher degrees of confidence result from smaller values
of N2.
In a given environment the velocity constant, k, for
05\
provides sufficient justification for its generalization
here. In logarithmic form, equation 3 may be written
as follows:
logk= - - /+logA (4)
For all practical purposes, log A may be treated as a
constant.2 Then equation 4 is of the form y = mx + b
2 The significance of the term, log A, is evident from exami-
nation of the Eyring rate equation. See Johnson et al. (1954).
It includes an entropy term, which in cases of spore de-
struction is quite large. Occasionally, values of AS along with
values of /Aare listed for thermal spore destruction. This com-
pletely defines the velocity constant, k, as a function of tem-
perature.
'OS

A) TEMPERATURE 220 F. B) TEMPERATURE 268.7 F.

REDRAWN FROM DATA REDRAWN FROM DATA
14 OF BALL (1943) 104 _ \OF STERN AND PROCTOR (1954)

-I
w

-J
103

k - 0.25
102 \ k . . 00057 SEC.-'I ,o
0
SEC.-'

z
z

* TIME MINUTES -0- TIME MINUTES

FIG. 1. Survivor curves for spores of Bacillus stearothermophilus strain 1518 at two different temperatures
1957] STERILIZATION OF NUTRIENT MEDIA 223

Injection of steam introduces some dilution to the

and a plot of log k versus for particular
spores
T
species in a given medium should yield a straight line
having a slope equal to 23RT* A typical plot is shown
-
in figure 2. Such plots are constructed by determining
the velocity constant, k, at at least two temperatures.
If there is doubt that the kinetics of the spore destruc-
tion are such that the velocity constants are not related
to temperature as in the Arrhenius equation, more k
values should be determined. The subsequent treat-
ment is not affected by another relationship, as long as
it is known. For spores of B. stearothermophilus strain
1518, characterized in figure 2, the activation energy
of a
medium, but this is taken into account during batch
make-up prior to sterilization.
Batch sterilization conditions are often specified as a
holding period at a certain temperature. The heat
effects involved during the time required to reach the
desired sterilizing temperature and the time required in
cooling down from this temperature are usually neg-
lected. In large-scale equipment, the rising and falling
temperature portions of the heating cycle are much
longer than the constant temperature portion.
Figure 4 compares actual fermentor heating cycles for
various sized vessels. Note that 110 F is the starting
temperature for the heating cycle. Batch make-up using

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for destruction is 67.7 Kcal/mole. Activation energies warm water can save considerable time in the solution
and entropies for thermal destruction of other bacterial
spores are listed in table 2.3

Batch Sterilization
The most common method of heat sterilizing nutrient
media is the batch method. The medium ingredients are
charged directly into the fermentor. The fermentor and
medium are sterilized by heat transferred across the -1 f

jacket and/or coil surfaces from condensing steam.

Fermentors are usually of the geometric design shown I.-
z
4n

in figure 3. The heat transfer surfaces are marked in z

figure 3 by heavy lines. The medium is agitated and 0

Fa
often steam is injected directly into the medium 3--
through the air sparger to speed up the sterilization. 0
-J
>
3Various other terms commonly are used to describe ther- 1i0
mal behavior of bacterial spores during heat sterilization,
especially in the food industry. The terms used in this article
are consistent with those used in chemical reaction kinetics.
Some other terms and their relationship to the velocity con-
stant, k, are tabulated for reference purposes.
0.00135 000137 0.00139 0.00141 0.00143 0.00145 0.00147 0.00149
Term Relation Definition RECIPROCAL TEMPERATURE R
T ~~~~~~~R
kT The ratio of the velocity constant at a FIG. 2. Effect of temperature on the velocity constant for
Qio destruction of spores of Bacillus stearothermophiluis strain
klTjo particular temperature to the veloc-
ity constant at a temperature ten 1518.
degrees lower. This ratio is often
falsely assumed constant over the TABLE 2. Activation energies and entropies for first order
entire temperature range. It dimin- degradation of B-complex vitamins and death of bacterial
ishes as temperature is raised spores
D 2.3 Two synonymous terms representing Energy Entropy
Vitamin or Bacterial Spore AS
k the time required for 90 per cent de- A
2.3 struction of a spore population at a cal/mole cal/mole K
k particular temperature, often called
the decimal reduction time Folic acid ............................... 16,800* -14.1 t
d-Panthothenyl alcohol ....... ........... 21,000* 5. it
Cyanocobalamin ............. ............ 23,100* 2.2t
TDT 2 3 log N, Thermal death time, a term attached
to the time required for "complete" Thiamine hydrochloride .................. 26,000* 11.4t
k N2 Bacillus stearothermophilus strain 1518.... 67,700 105
destruction of spores in a particular
environment. Algebraically, this Putrefactive anaerobe NCA 3679 ..... .... 72,400t 1234
Clostridium botulinum ......... ........... 82,l00t 160t
term is meaningless unless N2 has
some finite value *
Garrett (1956).
F as above The thermal death time at 250 F t Calculated from data of Garrett (1956).
t Levine (1956).
224 F. H. DEINDOERFER
of batch ingredients and in heating time. If water at
50 F were used in batch make-up it would have taken
an additional 30 min to heat the medium in the 15,000-
gal fermentor to 110 F. In order to avoid extending the
graphs unduly, 110 F was used as the finishing tempera-
ture. Actually, temperatures below 200 F have little
effect on the sterilization.
All the vessels characterized in figure 4 are reasonably
geometrically similar. Because of this congruency, the
heat transfer area per unit volume decreases as vessel
size increases. Also, since the vessels operate at similar
power input per unit volume levels, the heat transfer
coefficient also decreases as vessel size increases. This
[VOL. 5
explains the different shapes of the heating cycles in
figure 4. Coils are added to larger fermentors to provide
additional heat transfer surface. Very often, too, the
amount of steam directly injected into the medium
per unit volume is increased for larger fermentors. De-
spite this, however, the disadvantages of increased size
are not fully compensated.
Obviously, the rising and falling portions of the
sterilization cycle contribute significantly to fermentor
and medium sterilization, especially in larger vessels. A
method for determining the contribution of these
portions to the sterilization is suggested through the
use of the thermal-death relationships discussed earlier.

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RAW INGREDIENTS
WATER
ACID OR BASE

STEAM INILETS L {=
AND EXHAUST AIR OUTLET
COOLING WATER (STEAM VENT DURING STERILIZATION)
OUTLEETS

3 STEAM CONDENSATE OUTLETS

AND
3 COOLING WATER INLETS
STERILE AIR INLET C
($TEAM DURNIN STERILIZATION)
FIG. 3. Geometric design of large-scale fermentors

o TimE MINUTES * TIME MINUTES

FIG. 4. Temperature rising and falling curves during sterilization of medium in various sized fermentors
 STERILIZATION OF NUTRIENT MEDIA 225
The method involves a graphical integration of the presterilized medium. For how long must the medium
velocity constant over these portions of the cycle. k is a be sterilized at 250 F to be sure of sterilization in 999
function of T as shown in figure 2 and described in out of 1000 batches?
equation 3. T is a function of time, 0, as described by the Solution. For sake of illustration, assume the curve
heating cycle. Therefore, k is also a function of 0. An in figure 2 characterizes the most heat resistant bacterial
average value of k for each portion of the cycle can be spores in the penicillin medium. Also, let the heating
obtained by graphically integrating k over the time cycle of the 15,000-gal fermentor shown in figure 4
period involved and dividing the integral by the time represent the heating cycle of the fermentor under
represented by the portion, as in equation 5 below. consideration. 220 F is chosen as the minimum lethal
2 temperature. This is an arbitrary choice, of course.
lCv k dO (5) Actually lower temperatures are lethal but their
kavg =
1
relative lethality is small and neglecting them does not
02 - 01 introduce any significant error.
It should be kept in mind that the heating cycle Since the percentage of the laboratory bacterial

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rising and falling curves for a vessel will vary with a count that is contributed by the most heat resistant
number of factors. These include the liquid physical spores was not determined, assume that the entire
properties of the nutrient medium such as density, count was contributed by these spores. This is equiva-
viscosity, thermal conductivity, and specific heat; lent to adding a safety factor in the calculation.
extent of fouling of the heat transfer surfaces; amount Then
and enthalpy of steam sparged directly into the me-
N1 = (20 X 106 spores/ml)
dium; medium charge volume; temperature and (3.78 X 103 ml/gal)(12 X 103 gal)
enthalpy of the heating steam and temperature of = 9.07 X 1014 spores
the cooling water in the vessel coils and jacket. For a
given process and given vessel these variables are The chance for failure has been set at one in a thou-
maintained reasonably constant, and good replication sand so that N2 = 0.001 spore.
in heating cycles occurs. The velocity constants at the temperatures along the
The method is illustrated by way of the following rising portion of the sterilization cycle in figure 4 are
hypothetical example. determined from figure 2. These values are plotted
Example 1. A 15,000-gal fermentor containing 12,000 versus the time corresponding to the temperatures
gal of a penicillin production medium is to be sterilized. along the cycle as in figure 5A. The procedure is re-
The medium contains 4 per cent corn steep liquor. Corn peated for the falling portion of the cycle in figure 5B.
steep provides an excellent source of contaminating Note that the time of each of these portions below 220 F
organisms. Laboratory checks have shown bacterial is not shown on the figures. Graphical integration of the
counts not exceeding 20 X 106 cells per ml in the area under the curves in figures 5A and 5B, and solu-

.024 A) RISING PORTION OF CYCLE 8) FALLING PORTION

OF CYCLE
-10
I- .020
z

z
0
.016

0
-I
w .012

.008o

.004 f kd - o. 331 fkde 0.087

2kav0.335
9 0.0112 SEC.-l kav 0.0870,0066 SEC. 1
~~~~I i-31 0 I
______ I. I I I
0 5 10 15 20 25 0 S 10

0 TIME MINUTES

FIG. 5. Velocity constant variation over rising and falling temperature portions of sterilization cycle
226
tion of equation 5, yields values of kavg for each of these
portions of the cycle. For 29.5 min of the rising portion,
kavg is equal to 0.0112 sec-1 and for 13.2 min of the
falling portion, kavg is equal to 0.0066 sec-'.
Therefore, for portions of the heating cycle totalling
42.7 min, a kavg of 0.0098 sec-' can be used to calculate
the contribution of these portions to the sterilization.
Using equation 2, the population remaining if these
two periods followed each other can be calculated.
X 1014
log-9.07 N2
(0-0098 sec') (60 sec/min) (42.7 min)
2.3
F. H. DEINDOERFER [VOL. 5
Thus, only 8.8 min at 250 F are required for the
sterilization of the fermentor.
For comparison, similar calculations have been
made for the other sterilization cycles depicted in
figure 4. The results are listed in table 3. As vessel size
increases, the respective contributions of the rising and
falling portions of the cycle increase, and consequently
the time required at so-called sterilization temperature
decreases.
Heat Effects on Nutrients
The time required for batch sterilization is not
usually the optimum time of heating when product

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yield is considered. The thermal effects on nutrient
N2 = 1.14 X 10' spores quality of the medium must also be taken into account.
The N2 just calculated becomes the new N1 to be used Sometimes, productivity is increased by prolonged
in equation 2, this time to calculate the time the heating of the medium. In most media, however, the
fermentor must be held at 250 F to reduce the popula- deleterious effects of extensive heating are more ap-
tion to the desired N2 of 0.001. parent than any beneficial effects, and overheating of the
medium must be minimized.
2.3 1.14 X 103 An easily destroyed nutrient quality might be any one
(0.0265 sec-) (60 sec/min) log 1 X 10-3
0 =
0 of the B-complex vitamins. Vitamin degradation is
0 = 8.8 min known to occur rapidly at high temperatures. Al-
though figures for vitamin destruction in fermentation
media are not published, Garrett (1956) has studied
TABLE 3. Time at sterilization temperature to achieve the same
degree of sterilization in different sized fermentors vitamin stability at elevated temperatures in liquid
preparations. He found activation energies of from
Fermentor Size Total
CycleTime of Heating
Above 220 F Time at 250 F 16,800 to 26,000 calories/mole for destruction of several
gal m# mn B vitamins. The vitamins investigated by Garrett are
50 28.0 17.5 listed with their activation energies and entropies in
150 33.7 12.6 table 2. The activation energies and entropies for the
1,500 41.3 11.3 vitamins are much lower than for the three bacterial
15,000 51.5 8.8
spores also listed.
STERILE
RAW INGREDIENTS STEAM MEDIUM
WATER
ACID OR BASE

MAKE -UP
TANK

UNSTERILE I
MEDIUM WATER
FIG. 6. Steam-injection type of continuous sterilizer
1957] STERILIZATION OF NUTRIENT MEDIA
The conditions in a batch sterilization carried out at
250 F cannot always achieve sterilization without
impairing nutrient quality of the medium. Higher tem-
perature-shorter time batch sterilizations can be carried
out, but fermentation vessels are usually limited to
operation at not more than 30 psig (274 F) by design
restrictions.
Continuous Sterilization
A sterilization method becoming increasingly popular
and having several advantages over the conventional
batch sterilization is continuous sterilization. Contin-
uous sterilization of media for riboflavin fermentations
has been reported by Pfeifer et al. (1950) and for
penicillin fermentations by Whitmarsh (1954).
In one type of continuous sterilization, preheated
unsterile medium passes through an injection heater in
which steam is introduced. The vigorous and almost
instantaneous mixing obtained raises the medium to
sterilization temperature immediately. This tempera-
ture is maintained for the required amount of time in an
insulated retention tube through which the hot medium
flows. A diagram of a typical continuous sterilizer is
shown in figure 6. The hot medium passes through a
heat exchanger where it is cooled to below its flash point
as it preheats unsterile medium. Final cooling to process
temperature is accomplished in the fermentor.
The activation energy of bacterial spore destruction
is much higher than the activation energy of simpler
chemical reactions. It is, therefore, an advantage to use
a high temperature-short exposure time sterilization
operation whenever nutrient degradation occurs to the
extent that it lowers the process yield. Continuous
sterilization not only overcomes unfavorable nutrient
destruction, but has a number of operational ad-
vantages. Pfeifer and Vojnovich (1952) point out these
advantages in an excellent paper on continuous sterili-
zation. Operating conditions employed by these authors
for continuous sterilization of several types of media
are shown in table 4. In a batch sterilization, probably
TABLE 4. Operating conditions employed for
continuous sterilization of mnedia*
Media Used Suspended Solids pH Temra- Time
%t70 F min
Riboflavin ........... 1.8 corn steep liquor 4.5 275
Cyanocobalamin ..... 4.0 soybean meal 4.5 325
and distillers'
solubles
Acetone, butanol ...1 1.8 ground corn 6.5 275
Sodium gluconate 0.4 corn steep liquor 4.5 275
Itaconic acid ........0 .2 corn steep liquor 6.1 300
Fungal amylase...... 4.0 distillers' sol- 5.0 325
ubles and
3.0 ground corn
* Pfeifer and Vojnovich (1952).
227
only those sterilizations at 275 F could be approached,
if at all, in conventional fermentors.
The calculation of sterilization conditions for con-
tinuous operation is simplified by almost instantaneous
rising time to sterilization temperature and a rapid
cooling period thereafter. For this type of sterilization,
only the velocity constants of the most resistant spores
present in the medium and the total initial bacterial
concentration need be known to perform the calcula-
tion. Usually, in production operations the sterilizer is
of a fixed length. Different retention times are achieved
by controlled pumping. If temperature conditions are
specified, the retention time can be calculated easily
from equation 2. If process conditions dictate a certain
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flow rate, as may be the case in continuous fermenta-
tions, any modification in sterilizer operation will have
to be made temperature-wise. Consider the following
example.
Example 2. Process conditions are such that a reten-
tion time of 5 min is required in the sterilization of a
sodium gluconate production medium containing 0.4
per cent corn steep liquor. Plate counts of 3 X 106
bacteria per ml in the presterilized medium have been
determined in the laboratory. At what temperature
should the sterilizer be operated to achieve a steriliza-
tion having 99 per cent certainty of being successful?
One hundred thousand gallons of medium are to be
sterilized in this manner.
Solution. For sake of illustration, the curve in figure
2 again will be assumed as characterizing the most heat
resistant spores in the medium. The initial population
again will be assumed as entirely consisting of this
species.
Then
N1 = (3 X 106 spores/ml)
(3.78 X 103 ml/gal)(1 X 101 gal)
= 1.13 X 1015 spores
Since one chance in one hundred was chosen as the
margin of failure, N2 is equal to 0.01. Knowing the re-
quired retention time, equation 2 can be used to solve
for the necessary velocity constant.
2.3 1.13 X 1015
300 1 X 10-2
4 = 0.131 sec-1
13
The calculated value of k is used to read the required
sterilization temperature from figure 2. For k = 0.131
3
5 sec- 1 T = 0.1382. Thus, T = 724 R and a sterilization
5
13 temperature of 264 F is required.
To achieve the same sterilization at 250 F, a retention
time of 24.8 minutes would be required. Figure 7
illustrates the relationship between time and tempera-
228 F. H. DEINDOERFER [VOL. .5

ture to achieve the above sterilization. Also listed in Design Method

table 5 are the relative effects of an adverse chemical
reaction destroying a hypothetical vitamin during the
sterilization, the degradation of which is characterized
by an activation energy of 22.6 Kcal/mole, one-third
that of the activation energy for bacterial spore destruc-
tion. Even for the retention time and temperature used
in example 2, the vitamin was almost completely
destroyed. Much of the vitamin quality can be retained,
however, by higher temperature-shorter retention time
sterilizations.
The use of well-known thermal behavior characteris-
tics of bacterial spores and the temperature character-
istics of the sterilization cycle in calculating the time
for sterilization of fermentation medium has been
illustrated. This method offers an approach to the
correlation of sterilization conditions among various
sized fermentation vessels and between temperature
and retention time in continuous sterilizers. The advan-
tages of continuous sterilization have been pointed out
in a sterilization where nutrient damage occurs.
The steps involved in calculating process conditions
for sterilization operations can be summarized as
follows:

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(1) Periodically determine the thermal-death relation-
ship of the most heat resistant bacterial spore in the
10.2 medium to be sterilized.
(2) Determine the initial population concentration of
the medium and choose a confidence level for the
40
sterilization. An added safety factor is introduced by
assuming that the total population consists entirely
of the most heat resistant spores.
2
(3) Calculate the contribution to the sterilization of
the rising and falling portions of the sterilization cycle;
w
go-2 Il this step is unnecessary for steam injection type con-
tinuous sterilizations.
(4) Calculate the time required to hold the medium
isothermally at the highest temperature chosen for the
sterilization. For continuous sterilizations, the time of
o2~
exposure is often chosen, and calculations are carried
0~~~~~~~~~~~~~
out to find the required temperature.
REFERENCES
BALL, C. 0. 1943 Short-time pasteurization of milk. Ind.
0.00129 0.00131 0.00133 0.00135 0.0057 0.00139 000141 0.00143
Eng. Chem., 35, 71-84.
I
GARRETT, E. R. 1956 Prediction of stability in pharmaceu-
RECIPROCAL TEMPERATURE tical preparations. II. Vitamin stability in liquid multi-
FIG. 7. Time-temperature relationship to achieve the same vitamin preparations. J. Am. Pharm. Assn., 45, 171-178.
JOHNSON, F. H., EYRING, H., AND PILISSAR, M. J. 1954 The
degree of sterilization in a continuous sterilizer. Kinetic Basis of Molecular Biology, p. 220. John Wiley
& Sons, New York, N. Y.
TABLE, 5. Time-temperature relationship and its effect on vitamin LEVINE, S. 1956 Determination of the thermal death rate
content in a continuous steri,lization of bacteria. Food Research, 21, 295-301.
PFEIFER, V. F., TANNER, F. W., VOJNOVICH, C., AND TRAUF-
Relative Retention of LER, D. H. 1950 Riboflavin production by fermentation
Temperature Time Original Vitamin with Ashbya gossypii. Ind. Eng. Chem., 42, 1776-1781.
Content*
PFEIFER, V. F. AND VOJNOVICH, C. 1952 Continuous sterili-
F min % zation of media in biochemical processes. Ind. Eng.
250 24.8 0.0 Chem., 44, 1940-1946.
265 4.1 0.0 RAHN, 0. 1945 Physical methods of sterilization of micro-
280 0.72 2.3 organisms. Bacteriol. Reviews, 9, 1-47.
295 0.14 28 STERN, J. A. AND PROCTOR, B. E. 1954 A micro-method and
310 0.029 64 apparatus for the multiple determination of rates of de-
325 0.0061 89 struction of bacteria and bacterial spores subjected to
heat. Food Technol., 8, 139-143.
* k for vitamin destruction assumed equal to k for spore WHITMARSH, J. M. 1954 Continuous sterilization of fermen-
destruction at 250 F. tation media. J. Appl. Bacteriol., 17, 27.