Heat Sterilisation

Microbial

Heat Resistance & Survivor Curves;

Thermal Death times & Spoilage probability;
Process Calculations
Processing Systems
Dr. Ian Thompson

Preservation Processes
Definition: Processing steps required to reduce or eliminate the
potential for food borne illnesses and spoilage.
Pasteurisation (traditional):

Increase in product temperature to inactivate specific pathogenic

micro-organisms;

Is product Shelf stable?

Typically a few weeks with refrigeration (< 5 oC);

Commercial Sterilisation:
More intense thermal process to reduce population of all
microorganisms in product;
Is product shelf stable?

Absolutely, for 12 months or more at room temperature

(does not require refrigeration).

Microbial Survival Curves

The main issue relating to food preservation is the
microbial population;
An external agent (heat) is used to reduce the population
of microbes present;
Vegetative cells (eg. E. coli, Salmonella, Listeria monocytogenes) will
decrease;
Microbial spores will decrease in a similar manner after a
lag period;

Effect of Heat on Micro-organisms

Preservative Effect due to denaturation of Proteins:

Destroys enzyme activity & enzyme controlled metabolism in
microbes;

Rate of destruction is a first order reaction

When food is heated to a given temperature, the same % die in a
given time interval regardless of the initial numbers present.
This gives rise to a logarithmic order of death survival curve
(death curve);
dN/dt = - k.N n , n = 1 for first order;
N = microbial population, t = time, k = first order rate constant;

Microbial Survivor Curves

Decimal reduction time (D):

Time needed to destroy 90% of the microorganisms i.e. to
reduce the load by a factor of 10;
Higher D values means higher thermal resistance
Slope of death rate curve (log N vs. time) for 1 log reduction;
Consider:
At t = 0, initial population = No,
After time t, population = Nt.
Then: log No log Nt

(log No / log Nt) = c. t

By definition,
When (log No / log Nt) = 1, then t = tD & c = 1/D

(log No / log Nt) = t / D

(N/No) = 10 t/D

[Eqn. 1]

Problem 1
The following data were obtained from a thermal
resistance experiment conducted on a spore suspension
at 112 oC:
Time
No. of survivors
0
10 6
4
1.1 x 10 5
8
1.2 x 10 4
12
1.2 x 10 3
Determine the D value for the organism.
(Ans: 4.1 minutes)

First Order Kinetics &

Decimal Reduction Time

First Order Kinetics:

dN/dt = - k.N n , n = 1 for first order;
(N/No) = e kt

From Decimal Reduction time:

(N/No) = 10 t/D
On condition of a first order survivor curve:
10 t/D = e kt 2.303/D = k
[Eqn. 2]
First order rate constant (k) is inversely proportional to
decimal reduction time (D).

Commercial Application
Approaches to establish thermal process

Time/temperature combination dependent of m/b load.

Higher numbers of microbes will require longer time variation in
microbial load of raw material will require recalculating process for
each batch;
OR

Specific time/temperature combination used for every

batch irrespective of microbial load
Adequate preparation procedures are used to ensure that raw
material has acceptable and uniform quality (specifications);

Commercial Application
Microbial Destruction occurs logarithmically

Is it possible to destroy all microorganisms?

By heating for an infinite time since population will only approach
zero;

From survivor curve equation:

N = N0 x 10-(t/D) N0 only if t
[Eqn. 3]
An infinite time will be required for the destruction of all
viable microorganisms.
OR
Reduce microbial load by predetermined amount or factor
Basis for concept of commercial sterility.
The probability of survival of a single micro-organism after heat
treatment can be predicted based on microbial loads, heat
resistance, temperature and time of heating.

Thermal Death Time Curve

The destruction of microbes is temperature

dependent
More cells die at higher temperature
By collecting D-values at higher temperatures, a
thermal death time curve (TDT) may be
constructed;
The slope of the TDT curve is the number of degrees
required to effect a 10-fold change in D = z value;

Thermal Death Time Curve

The thermal resistance
constant (z) describes the
influence of temperature (T)
on the decimal reduction
time (D);
z = change in T (T2 T1) [Eqn. 4]
log D1 logD2
Hence, the D value (time)
and z value (temperature) is
used to characterize the heat
resistance of microbes;

Problem 2
The decimal reduction times D for a spore suspension
were measured at several temperatures, as follows:
Temp (oC)
104
107
110
113
116

D (min)
27.5
14.5
7.5
4.0
2.2

Determine the thermal resistance constant z for the spores.

(Ans. = 11 oC )

Heat Processing data

Factors affecting the Heat

Resistance of Microbes

Type of Microbes
Different species/strains show wide variation in heat resistance;
Spores are more heat resistant than vegetative cells

Incubation conditions (during cell growth or spore formation)

Temperature spores produced at higher temperature are more
heat resistant;
Age of vegetative cells;
Medium of growth (fatty acids influence heat resistance);

Conditions during heat treatment

pH of food (pathogenic/spoilage bacteria more heat resistant at
neutral pH

Factors affecting the Heat

Resistance of Microbes

Conditions during heat treatment

pH of food (pathogenic/spoilage bacteria more heat resistant at
neutral pH: yeast and fungi less heat resistant than bacteria)
Water activity aw

Moist heat - more effective than dry heat;

Composition of food (protein/fat/sucrose cause an increase);

Growth media and incubation conditions

Used to assess recovery (survivors) of microbes in heat
resistance studies;

Microbes vs Enzymes
Most enzymes have D & z values within a similar range
to microbes;
Some enzymes are very heat resistant and may not be
denatured by relatively short heat treatments
The heat resistance of enzymes and microbes found in
specific foods is used to calculate the heating conditions
needed;
In practice, the most heat resistant species
(microbe/enzyme) is used a basis for calculating the
process time/temperature conditions.

Thermal Death Time (F)

Total time required to accomplish a stated reduction in a
population of vegetative cells or spores;
Expressed as a multiple of D-values (on a 1st order survivor
curve model);
Therefore, 4 D = 10 -4 reduction = 99.99% reduction
For commercially sterile, low acid shelf stable foods,
thermal death time (F) = 12D.
Reference thermal death times = Fo (sub-zero) - where process
temperature and z values are specified;
Eg. For C. botulinum, temperature = 121.1 oC & z value =10oC;

- i.e. F Tz

or

10
121

Spoilage Probability

The spoilage probability is used to estimate the number of spoiled

containers within a total batch of processed product.
log No - log N = t / D;
If No & N represents the initial & desired final microbial
population respectively - for a thermal death time F, then

RECALL:

log No - log N = F / D.

If the total number of containers being processed = r, then the

total microbial load at the beginning of the process =
r x No
and
log (r. No) - log (r. N) = F / D
If the goal is to achieve a probability of one survivor from all
containers, then log (r. N) 0 and
log (r. No) = F / D r. No = 10 F/D
the spoilage probability, 1/r = No / 10

F/D

Spoilage Probability
1/r = No / 10 F/D

[Eqn. 5]

1/r - represents the total number of containers processed

(r) resulting in 1 with spoilage;
The expression can be used to estimate F, given No & D.

Prevailing assumption:
The survivor curve for the spoilage microorganism is 1st order.

Problem 3
Estimate the spoilage probability of a 50 minute process at
113 oC when D = 4 minutes, and the initial microbial
population is 10 4 per container.

Solution:
1/r = No / 10 F/D
= 10 4 / 10 50/4 = 10 4 / 10 12.5
= 10 -8.5 = 3.16 x 10 -9.
r = 3.16 x 10 8 - i.e. the spoilage of 1 container in 3.16 x 10 8
containers (r) can be expected.

General Method for

Process Calculations

Based on classical paper by Bigelow (1920);

Major Assumption:
The thermal death time, F, for the microbial population considered must
be known at all temperatures to which the product is exposed during the
preservation process.
Recall: Thermal Death Time, D, decreases with increasing temperature

Method:
1. Draw sterility curve for process: Sterilization rate (F/t) vs. time
2. Area under the curve (in time units) is the lethal effect of
the process (lethality) i.e. the integrated impact of
time and temperature on the microbial population.

General Method for

Process Calculations
Recall: relationship with Thermal Death time (F)
log No - log N = F / D.
& Thermal resistance constant, z
z = (T2 T1)
log D1 logD2
Derived to give:
log FR log F = (T TR) FR / F = 10 (T- TR)/z
z
-

[Eqn. 6]

where FR is the thermal death time known at a reference temp, TR

This eqn. can be used to compute the thermal death time, F at any
temperature,T .

Application to Pasteurisation

During batch pasteurisation, the food is heated to a

defined temperature and held for a defined period to
achieve a certain lethality.
Only the holding period is considered (not the heating
or cooling phases);
Pasteurisation of milk is based on the reduction of a
microbial pathogen with:

D 63 = 2.5 min, z = 4.1 oC, & a thermal death time, F = 12 D.

Process ensures survival of pathogen is negligible.
The traditional batch process is holding at 63oC for 30 min.
Since the reference TR is 63oC, the lethality is 1.

Application to Pasteurisation

The continuous HTST process attains lethality during a

holding period at a temperature close to the heating
medium;
The extent to which heating and cooling will contribute to
lethality depends on the rate of heating and cooling;
For HTST, the milk product is heated to 71.5 oC and it will
only need to be held for 15 sec. to achieve the same
lethality (=1) as the batch process at TR.
If it were to be held for 30 minutes, the lethality would be
120 (but the milk would be no good).

Problem 4

A thermal process is accomplished by instantaneous

heating to 138 oC followed by a 4 sec hold and
instantaneous cooling.
Estimate the lethality (in sec) at 121 oC if the thermal
resistance of the microorganism is 8.5 oC.
Hints:
use eqn - FR / F = 10 (T - TR)/z
TR = 138 oC & FR = 4 s
F121 = F138 x 10 (138 121)/ 8.5

= 4 x 10 2 = 400 s

Reference Material
Prescribed Text:
Fellows, Peter J., (2009). Food Processing Technology:
Principles and Practice, 3rd edition. Woodhead: Cambridge,
England. ISBN 1-4398- 0821X

Highly Recommended Reading:

Singh, R Paul & Heldman, Dennis R., (2009). Introduction to
Food Engineering. 4th edition. Academic Press: Amsterdam.
ISBN 0-1237-370900-4.
Online Resources
Earle, R.L. and Earle, M.D., Unit Operations in Food Processing
(Web edition), New Zealand Institute of Food Science and
Technology. http://www.nzifst.org.nz/unitoperations/index.htm;