”Mikrobiologiske aspekter af levnedsmiddelkonservering og kvalitetssikring”

16 March 2005 (course 27751 / 076018)

Predictive microbiology

Paw Dalgaard
Microbiology group
Danish Institute for Fisheries Research (DIFRES)
Dept. of Seafood Research (FF)
DTU, Lyngby building 221
www.dfu.min.dk/micro/pd.htm
pad@dfu.min.dk
DFU, Lyngby FF

Predictive microbiology – course 27751 / 076018

(Predicting shelf-life and safety of foods)

• Relation to other courses

• Summary of previous lactures re. predictive microbiology
• Primary growth models
• Logistic models
• Secondary growth models
• Cardinal parameter models (CPM)
• Growth/no-growth interface models
• Exercises with ComBase, MicroFit ans SSSP software

DFU, Lyngby FF

1
 Predictive microbiology – course 27755 / 076018
Relation to other courses include predictive microbiology

• Levnedsmiddelmikrobiologi (071213):
• Koncept og oversigt over tilgængelige modeller og software
(MicroFit, ComBase, Growth Predictor, PMP, SSSP)

• Fisk som råvare (27501):

• Ferske fiskeprodukter - forudsigelse af holdbarhed

• Fisk som levnedsmiddel og ressource (27502):

• Letkonserverede fiskeprodukter
• Forudsigelse af holdharhed og sikkerhed
• Kvadratrodsmodeller
DFU, Lyngby FF

Predictive microbiology – course 27755 / 076018

Recommended literature about predictiv microbiology:

McKellar, R. C. and Lu, X. (2004). Modeling Microbial Responses in

Foods. CRC Press, Boca Raton, Fl, USA. p. 343.

Legan, D., Vandeven, M., Stewart, C., Cole, M. (2002). Modelling the
growth, survival and death of bacterial pathogens in foods. (Chapter 3)
In: Foodborne pathogens. Blackburn, C. de W. and McClure, P.J. (eds.).
Woodhead Publishing Ltd. pp. 53-95.

DFU, Lyngby FF

2
 Predictive microbiology – the concept

• Growth, survival and inactivation of microorganisms in foods are reproducible

responses

• A limited number of environmental parameters in foods determine the kinetic

responses of microorganisms
• Temperature
• Water activity/water phase salt
• pH
• Food preservatives (organic acids, nitrite, …)

• A mathematical model that quantitatively describes the combined effect of the

environmental parameters can be used to predict growth, survival or inactivation of
a microorganism and thereby contribute important information about product
safety or shelf-life

DFU, Lyngby Roberts and Jarvis (1983) FF

Development of predictive microbiology models

Models are usually developed in two steps from large experiments
including the effect of several environmental parameters
10
9
8
Growth
7 rate
Log (cfu/g)

6
5
4
3
2
Lag time
1
0
Storage time

Primary model Secondary model

Models allow microbial responces to be predicted at conditions
DFU, Lyngby that have not been specifically studied FF

3
 Development of predictive microbial models

Data generation : Growth curves are generated in model systems for combi-
nations of environmental factors (temp., pH, NaCl, etc.)

Primary modelling : Growth curves are fitted by sigmoidal growth models

Secondary modelling : The effect of controlling factor(s) on kinetic parameters

(e.g. the lag phase and the growth rate) is modelled

Product validation : Predicted values of kinetic parameters are compared to

values obtained in products and challenge tests

Tertiary ‘modelling’ : Validated models are included in application software

DFU, Lyngby FF

Primary growth models

11
10
9
8
Log (cfu/g)

7
6
Exponential model
5
Logistic model without lag
4
3 Logistic model with lag
2 Baranyi & Roberts (1994)
1
0
0 50 100 150 200 250 300 350 400 450

Storage period (hours)

DFU, Lyngby FF

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 Secondary predictive microbiology growth models

Kinetic models
• Polynomial and constrained linear polynomial models
• Square-root-type models
• Cardinal parameter models
• Artificial neural networks
Growth/no growth interface models

DFU, Lyngby FF

Evaluation of secondary predictive microbiology models

Secondary predictive models are evaluated/validated by comparison of
measured/observed and predicted values (kinetic parameters or shelf-
life). The comparison can be graphical or mathematical
∑ log( µ − predicted / µ −observed )

• Bias factor (Bf) = 10 n

∑ log( µ − predicted / µ −observed )

• Accuracy factor (Af) = 10 n

• Bias- og accuracy- factors can be calculated from lag times, growth

rates, time for 1000-fold increase in cell concentration etc.
• Bf indicates if a model systematically predicts growth to be faster
(Bf > 1) or slower (Bf < 1) than observed in foods
DFU, Lyngby Ross (1996); Baranyi et al. (1999) FF

5
 Evaluation of Listeria monocytogenes growth models – seafood example

Non cured products Cured products

n Bias factor n Bias factor
Pathogen Modeling Program 15 2.2 26 3.9

Food MicroModel 28 1.4 60 2.3

Murphy-model 27 1.1 50 2.3

Ross-model 29 1.0 60 1.4

Acceptable model: 0.75 < Bias factor < 1.25

• Users of predictive microbiology models should assure models have been
successfully validated in a foods with microbial ecology similar to the product
of interest

DFU, Lyngby Dalgaard & Jørgensen (1998) FF

Application of predictive microbiology models

• Predict the effect of product characteristics and storage

conditions on microbial responses Æ safety and shelf-life
• Predict effect of changes in foods Æ product development
• HACCP plans – establish limits for CCP
• Food safety objectives – equivalence of processes
• Education – easy access to information
• Quantitative microbiological risk assessment (QMRA)
The concentration of microbial hazards in foods may increase or decrease
substantially (millions of folds) during processing and distribution

DFU, Lyngby Roberts and Jarvis (1983), Legan et al. (2002) FF

6
 Application of predictive microbiology in QMRA

Prevalence and Product Storage Storage time

conc.of hazard characteristics conditions (shelf-life)

Predictive microbiology model(s)

assessment
Exposure

(Growth limits model) + Kinetic model

Model = deterministic + stochastic part

Output: Predicted concentration of hazard in

food at the time of consumption

Predicted probability Consumption

charaxrerization

of illness per meal patterns

Hazard

Cases per 1 million Cases per 100.000

meals (sub)-population
DFU, Lyngby FF

Application of predictive microbiology models

1. Determine product characteristics and storage conditions of food

Temperature, aw/NaCl, pH, organic acids, nitrit,
smoke components, inhibting microflora
2. Secondary model → lag time, growth rate, etc.
3. Primary model → Growth curve (Concentration over time)

• Application software facilitates step 2 and 3

• Predictions can be useful or misleading depending on:
- Successful product validation and correct use of models
- Appropriate information about food and storage conditions
DFU, Lyngby FF

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 Predictive microbiology freeware applications
• Pathogen Modeling Programme (USA) - www.arserrc.gov/mfs/pathogen.htm
• 37 models of growth, survival and inactivation
• Frequently updated (version 7.0)
• Available free of charge during the last 15 years
• ~ 5000 downloads per year

• Growth Predictor (UK) - www.ifr.ac.uk/Safety/GrowthPredictor/

• Based on data previously used in the FoodMicromodel software
• 18 models for growth of pathogenic bacteria
• Available free of charge since 2003 (> 600 users)

• Seafood Spoilage and Safety Predictor (DK) – www.dfu.min.dk/micro/sssp/

• Time/temperature integration
• Shelf-life, specific spoilage organisms, Listeria monmocytogenes
• Available free of charge since 1999 (> 1200 users from > 50 countries)
DFU, Lyngby FF

Predictive microbiology application software

• ComBase (UK, USA) – wyndmoor.arserrc.gov/combase/

• Database with information on growth and inactivation of microorganisms
• ComBase containes ~35000 growth/inactivation curves in spring 2005
• Available since 2003

DFU, Lyngby FF

8
 Predictiv microbiology – course 27751 / 076018
(Predicting shelf-life and safety of foods)

• Relation to other courses

• Summary of previous lactures re. predictive microbiology
• Primary growth models
• Logistic models
• Secondary growth models
• Cardinal parameter models (CPM)
• Growth/no-growth interface models
• Exercises with ComBase, MicroFit, SSSP and PMP software

DFU, Lyngby FF

Primary growth models

10
9
8 Curve fitting freeware:
Growth
7 rate
Log (cfu/g)

6 MicroFit v. 1.0 is available free of

5 charge and can be used to estimate
4 kinetic parameter values from
3 growth curve data
2 (www.ifr.bbsrc.ac.uk/MicroFit)
Lag time
1
0
Storage time

N Cell concentration (cfu/g)

dN/dt Absolute growth rate (cfu/gram/hour)
(dN/dt)/N = µ Specific growth rate (1/hour)

DFU, Lyngby FF

9
 Primary growth models

Exponential model Log ( N t ) = Log [ N o × exp( µ max × time )]

Logistic model N max

Log ( N t ) = Log ( )
without lag ⎡ N max ⎤
1+ ⎢ − 1⎥ × exp(− µ max × time)
⎣ N0 ⎦

Logistic model N max − N min

Log( N t ) = Log( N min + )
with lag 1 + exp(−µ max (time − ti ))

Baranyi & Roberts (1994) ⎛

⎜
⎛ ⎡ 1 ⎛ exp( − µ max × time ) + q 0 ⎞ ⎤ ⎞⎟ ⎞⎟
exp ⎜ µ max × ⎢time + × Ln ⎜⎜ ⎟⎟ ⎥ − 1
⎡ ⎛ exp( − µ max × time ) + q 0 ⎞⎤ ⎜ ⎜ µ max ⎝ 1 + q0 ⎠ ⎦ ⎟⎠ ⎟
1 1 1 ⎝ ⎣
Log ( N t ) = Log ( N 0 ) + × ⎢time + × Ln ⎜⎜ ⎟⎟ ⎥ − × Ln ⎜1 + ⎟
µ max ⎣ µ max ⎝ 1 + q0 ⎠ ⎦ Log (10 ) ⎜ exp( Log ( N max ) − Log ( N 0 )) ⎟
⎜ ⎟
⎜ ⎟
⎝ ⎠

Modified Gompertz ⎛ ⎡ µ × exp(1) ⎤⎞

Log ( N t ) = Log ( N 0 ) + ( A × exp⎜⎜ − exp⎢ max × (lag − time) + 1⎥ ⎟⎟) / Ln(10)
model ⎝ ⎣ A ⎦⎠

DFU, Lyngby McKellar and Lu (2004) FF

Primary growth models

11
10
9
8
Log (cfu/g)

7
Exponential model
6
5 Logistic model without lag
4 Logistic model with lag
3
Baranyi & Roberts (1994)
2
Modified Gompertz model
1
0
0 50 100 150 200 250 300 350 400 450

Storage period (hours)

DFU, Lyngby FF

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 Primary growth models
The modified Gompertz model overestimates the maximum specific growth rate (µmax) by
~ 15 % (Maximum specific growth rate: µmax = Ln(2)/generation time)

Original Gompertz model

9
8
7
Nt = N max × exp( − exp( µ' (time − t i )))
6
Log(cfu/g)

5
Log ( N t ) = Log ( N max × exp(− exp(µ' (time − t i ))))
4
3
2
1
0
0 5 10 15 20
Storage period (days)

⎛ ⎡ µ × exp(1) ⎤⎞
Modified Gompertz model Ln( N t / N 0 ) = A × exp⎜⎜ − exp⎢ max × (lag − time) + 1⎥ ⎟⎟
⎝ ⎣ A ⎦⎠

DFU, Lyngby Turner et al. (1976), Zwietering et al. (1990) FF

Primary model for microbial interaction

8 LAB L. monocytogenes
7

6
log(cfu/g)

5
4
L. monocytogenes + LAB
3

2 L. monocytogenes with
1 lag phase + LAB
0
0 10 20 30 40 50 60
Days at 5°C

DFU, Lyngby Giménez & Dalgaard (2004) FF

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 Primary model for microbial interaction
• Jameson effect:
All microorganisms in a food stop growing when the dominating
microflora reaches its maximum population density

• Differential form of Logistic model for growth of LAB – intra-species competition

9

dLAB/ dt ⎛ LABt ⎞ 8
= µ max
LAB
× ⎜⎜1 − ⎟⎟ 7
LABt ⎝ LABmax ⎠ 6

-1
Log cfu g
5
4
3
2
dLm / dt ⎛ Lmt ⎞ ⎛ LABt ⎞
= µ × ⎜⎜1 − ⎟⎟ × ⎜⎜1 − ⎟⎟
Lm 1
max
⎝ max ⎠ ⎝ max ⎠
0
Lmt Lm LAB 0 1 2 3 4 5 6 7 8
Storage period (days at 25°C)

• Logistic model for growth and interaction between LAB and L. monocytogens (Lm)

DFU, Lyngby Giménex & Dalgaard (2004) FF

Predictiv microbiology – course 27751 / 076018

(Predicting shelf-life and safety of foods)

• Relation to other courses

• Summary of previous lactures re. predictive microbiology
• Primary growth models
• Logistic models
• Secondary growth models
• Cardinal parameter models (CPM)
• Growth/no-growth interface models
• Exercises with ComBase, MicroFit, SSSP and PMP software

DFU, Lyngby FF

12
 Cardinal parameter models

1.2 1.2
1.1 µopt 1.1 pHopt
1.0 1.0
0.9 Topt 0.9
-0.5

-0.5
0.8 0.8
Sqrt(µmax), h

Sqrt(µmax), h
0.7 0.7
0.6 0.6
0.5 0.5
0.4 0.4
0.3 0.3
0.2 0.2
0.1 0.1
Tmin Tmax pHmin pHmax
0 0
0 5 10 15 20 25 30 35 40 45 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5

Temperature (°C) pH

DFU, Lyngby Rosso et al. 1995 FF

Cardinal parameter models

2
⎛ Lactic acid ⎞
µ max = µ opt ⋅ CM 2 (T ) ⋅ CM 1 ( pH ) ⋅ CM 2 (a w ) ⋅ ⎜1 − ⎟
⎜ MIC Lactic acid ⎟
⎝ ⎠
MIC : ‘Minimal inhibitory concentraion’

⎧ 0, , X ≤ Xmin
⎪⎪ (X − Xmax) ⋅ (X − Xmin)n
CMn (X) = ⎨ n−1
, Xmin < X < Xmax
⎪(Xopt − Xmin) ⋅[(Xopt − Xmin) ⋅ (X − Xopt) −(Xopt − Xmax) ⋅ ((n −1) ⋅ Xopt + Xmin − n⋅ X)]
⎪⎩ 0, , X ≥ Xmax

Extensive cardinal parameter models including the effect of various inhibitory

components have been developed e.g. for Listeria monocytogenes

DFU, Lyngby Rosso et al. 1995, Augustin & Carlier 2000 FF

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 Lag time models

• Secondary lag time models can be developed in the same way as

growth rate models (1/lag time = lag rate)
• Lag time of microorganisms depend not only on environmental
parameters but also on the physiological state of the microorganisms
• Lag times data is more variable than growth rate data
• ’Relative lag times’ (RLT) = Lag time/generation time (tgen) are
less variable and used to predict lag time from µmax

Lag time = RLT ⋅ t gen = RLT ⋅ Ln(2) / µ max

DFU, Lyngby Ross and Dalgaard 2004 FF

Examples of available secondary models

No. of
Pathogens Independent variables and ranges
models
Aeromonas hydrophila 5 T(4-42°C); NaCl (0.5-4.5 %); pH (5.0-7.3); Na-nitrite (0-200 ppm); aerobic, anaerobic
Aspergillus spp. 4 T (25-37°C); aw (0.83-0.99); pH(6.5); humectant: glucose/fructose; propionic acid (129-516 ppm)
Bacillus cereus 5 T(5-42°C); NaCl (0.5-10.5 %); pH (4.5-7.5); Na-nitrite (0-200 ppm); aerobic; CO2 (10-80%); humectants:
NaCl, glycerol
Clostridium botulinum 5 T(4-30°C); NaCl (0.0-5.0 %); pH (5.0-7.3); CO2 (0-90%); Na-pyrophosphate (0-3%); Initial spore conc. (-2 -
5 log cfu/g); Initial aerobic plate count (-2 - 3 log cfu/g)
Clostridium perfringens 3 T(12-42°C); NaCl (0-3 %); pH (5.5-7.0.)
Escherichia coli 7 T(5-47.4°C); NaCl (0.5-15 %); pH (3.5-8.5); Na-nitrite (0-200 ppm); lactic acid (0-500 mM); oregano
essential oil (0.0-2.1%); reuterin (0-4 AU/ml); aerobic and anaerobic
Listeria monocytogenes 13 T (-2.7 – 47.4 °C); NaCl (0-19 %); aw (0.910-0.997); pH (3.5-9.61); acetid acid (0-20.1 mM); lactic acid (0-
5.4 mM); citric acid (0-1.6 mM); Na-benzoate (0-0.7 mM); K-sorbate (0-5.1 mM); sodium nitrite (0-11.4
µM); glycerol monolaurin (0-118.5 ppm); butylated hydroxyanisole (0-254 ppm); butylated hydroxytoluene
(0-48.7 ppm); tertiary butylhydroquinone (0-1400 ppm); methyl paraben (0-0.2%); CO2 (0-1.64 proportion);
caffeine (0-10.8 g/l); phenol (12.5 ppm); reuterin (0-4 AU/ml); curvaticin 13 (0-160 AU(ml); oregano
essential oil (0.0-2.1%); aerobic and anaerobic; smoke components/phenol (3-10 ppm); interaction with lactic
acid bacteria
Salmonella 6 T(5-48°C); NaCl (0.5-4.5%); pH (4.3-7.4); previous pH (5.7-8.6); oleuropein (0-0.8%); oregano essential oil
(0.5-2.0%); aerobic
Shigella 2 T(10-37°C); NaCl (0.5-5.0 %); pH (5.0-7.5); Na-nitrite (0-1000 ppm); aerobic
Staphylococcus aureus 4 T(5-45°C); NaCl (0-16.5%); pH (4.5-9.0); acidulants HCl, acetic acid or lactic acid; Na-nitrite (1-200 ppm);
aerobic and anaerobic
Vibrio parahaemolyticus 1 T(8-45°C); aw (0.936-0.995); aerobic
Yersinia spp. 4 T(1-42°C); NaCl (0.5-6.5%); pH (4.0-8.5); Na-nitrite (0-200 ppm); aerobic; CO2 (0-100 %); O2 (0-60 %)

DFU, Lyngby Ross & Dalgaard (2004) FF

14
 Predictiv microbiology – 27751 / 076018
(Predicting shelf-life and safety of foods)

• Relation to other courses

• Summary of previous lactures re. predictive microbiology
• Primary growth models
• Logistic models
• Secondary growth models
• Cardinal parameter models (CPM)
• Growth/no-growth interface models
• Exercises with ComBase, MicroFit, SSSP and PMP software

DFU, Lyngby FF

Growth/no growth interface models

National and EU legislation has stimulated interest in these models –
example for Listeria monocytogenes

Product type Number Number of m M

of samples positive samples cfu/g cfu/g
(n) between m and M
Lightly preserved without heat treatment
Allows growth of L. monocytogenes 5 0 0* -

Lightly preserved without heat treatment

Stabilized against growth of L. 5 1 10 100
monocytogenes

* Not detectable in 25 g

DFU, Lyngby FF

15
 Growth/no growth interface models
• The combinations og environmental parameters that prevent microbial
growth is important e.g. for product formulation (Hurdle concept,
Multiple barrier technology)
• Growth/no growth interface ( or growth limits) models include:
• Polynomial models
• Square-root type models
• Cardinal parameter model
• Logistic regression models
• Artificial neural networks (ANN)

• Small changes in environmental parameters causes shifts from growth to

no growth conditions and only few studies have compared the ability of
different models to predict the interface

DFU, Lyngby Ross & Dalgaard (2004) FF

Growth/no growth data for E. coli

0.99

Logistic
regression
0.98

Cardinal
parameter
0.97
model

0.96
ANN

0.95

CPM
0.94
0 5 10 15 20 25 30 35 40 45 50
Temperature ( C)

DFU, Lyngby Ross & Dalgaard (2004) FF

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 A cardinal parameter growth model predicted the growth/no growth
interface for Listeria

µ =µ ⋅ CM 2 (T ) ⋅ CM 1 ( pH ) ⋅ ξ (T , pH )
max opt

⎧ 1 ,ψ ≤ ½
⎪
ξ (ϕ (T , pH ) = ⎨2(1 − ψ ) ,½ < ψ < 1
⎪ 0 ,ψ ≥ 1
⎩
No interaction
ϕe
between T and pH ψ =∑ i

i 2∏ (1− ϕ e j )
j≠ i

Growth/no
growth interface
ϕT = (1− CM2 (T))2

ϕpH = (1−CM1( pH))2

DFU, Lyngby Le Marc et al. (2002) FF

Predictiv microbiology – course 27751 / 076018

(Predicting shelf-life and safety of foods)

• Relation to other courses

• Summary of previous lactures re. predictive microbiology
• Primary growth models
• Logistic models
• Secondary growth models
• Cardinal parameter models (CPM)
• Growth/no-growth interface models
• Exercises with ComBase, MicroFit, SSSP and PMP software

DFU, Lyngby FF

17
Exercise 1: How to obtain microbial growth data from ComBase and estimate
growth kinetic parameters by using MicroFit

Before you start the exercise:

Download MicroFit v. 1.0 from www.ifr.bbsrc.ac.uk/MicroFit and install this curvefitting
freeware on your PC. Also make sure you have MS Excel activated as well as access to
the internet.
Exercise: How much CO2 is required to significantly reduce the maximum specific growth
rate of Listeria monocytogenes at 5.0°C, pH 5.65-5.75 and water activity above 0.995?
-Access ComBase at http://wyndmoor.arserrc.gov/combase/, select ’Start search’ and enter
the search criteria (Temperature, pH, water activity, Condition = CO_2, Listeria
monocytogenes/innocua and Environment = culture medium). This search should result in
12 records/growth curves for Listeria monocytogenes Scott A.
- For these data ComBase does not provide the maximum specific growth rates or
doubling times. However, growth curve data is provided in ComBase and they can be
pasted e.g. into Excel or saved directly as csv-files (Comma Separated Variables) and
further edited. e.g. by MS Excel.

DFU, Lyngby FF

Exercise 1 continued: - To facilitate this exercise csv-files from Combase has been edited
in such a way that MicroFit can read the data and extimate kinetic parameters.
- Start MicroFit and estimate maximum specific growth rates for the 8 growth curves
included as csv-files in Exercise1.zip (this file can be downloaded from
www.dfu.min.dk/micro/pd.htm). Exercise1.zip also includes a lst-file with information
about the pH, %CO2, %O2 and %N2 for each of the 8 growth curves. To access the lst-file
from MicroFit use ’File’, ’Browse data set 1…’ and select ’List files (*.lst)’. For MicroFit
to read a growth curve use ’File’, ’Load data set …’ and ’Åbn’.
- When data are uploaded use the ’Fit Model’ bottom in MicroFit to estimate growth
kinetic parameters. MicroFit uses a modification of the four-paramater Logistic model
(Baranyi & Robewrts 1994) as primary growth model.
- Microfit allows two growth curves to be compared and it can be tested if. e.g maximum
specific growth rate (mumax) differ significantly. Use this facility to compare maximum
specific growth rates of Listeria monocytogenes Scott A growing with (i) 0% CO2 or
20.5% CO2; (ii) 20.5% CO2 or 41.5% CO2 and . (ii) 0% CO2 or 41.5% CO2.
- Is oxygen CO2/O2 gas mixtures increasing or reducing the maximum specific growth rate
of Listeria monocytogenes as compaed to CO2/N2 gas mixtures ?
DFU, Lyngby FF

18
 Exercise 1: Effect of CO2 on he maximum specific growth
rate (µmax) of Listeria monocytogenes
0.40
: CO2/N2 gas mixtures
0.35 : CO2/O2/N2 gas mixtures

0.30
µmax (1/day)

0.25

0.20

0.15
(37% O2)
0.10
(56% O2)
0.05

0 10 20 30 40 50 60 70 80

DFU, Lyngby % CO2 FF

Exercise 2: Predicting the effect of hygiene, temperature and atmosphere on shelf-

life of fresh MAP salmon using the Seafood Spoilage and Safety Predictor (SSSP)
software
Before you start the exercise: Download SSSP v. 1.0 form www.dfu.min.dk/micro/sssp/
and install this freeware on your PC. If required you can also download and install the
Microsoft.net Framework 1.0 from the same site
Exercise: Can modified atmosphere packaging with increased levels of CO2 compensate
for (i) higher initial concentrations of spoilage bacteria or (ii) moderately higher
temperature during storage ?
1) Start SSSP, select ’Microbial spoilage models (MSM)’, select ’Photobacterium
phosphoreum’ and activate the model ’Fresh MAP salmon steaks’
2) Predict shelf-life and maximum specific growth rate using an initial cell density of 10
cfu/g, 2°C and 0% CO2. The results should be 9.6 days and 0.081 1/hours:
- With an initial cell density of 1000 cfu/g the shelf-life is reduced to 7.2 days. Is it
possible to compensate for the higher initial concentration of P. phosphoreum by
increasing the concentration of CO2 in the modified atmosphere? If yes which equilibrium
concentration of CO2 results in a shelf-life of 9.6 days at 2°C and with 1000 cfu/g?
DFU, Lyngby FF

19
2) continued
- After packaging, CO2 is absorbed into the water- and lipid-phases of salmon and the
initial gas mixture used when packaging fresh MAP salmon must contain a higher
concentration of CO2 than the desired equilibrium concentration. Storage temperature,
initial gas/product ratio and intial CO2 concentration determine the equilibrium
concentration of CO2 in fresh MAP fish. Use the SSSP function ’Calculation of % CO2 in
fresh MAP fish’ to determine one or several combinations of initial gas/product ratio and
intial CO2 concentration that results in a CO2 concentration of 49% at 2°C.
3) With an initial cell density of 10 cfu/g for P. phosphoreum the shelf-life of fresh MAP
salmon in 0% CO2 is 9.6 days at 2°C but only to 5.9 days at 5°C. Is it possible to
compensate for the higher storage temperature by increasing the concentration of CO2 in
the modified atmosphere? If yes indicate one or several combinations of initial gas/product
ratio and intial CO2 concentration that can be used.

OBS. You can use the help-function within SSSP to obtain information about the models
used to predict growth of P. phosphoreum and absorbsion of CO2 into fresh fish.

DFU, Lyngby FF

Referencer 1
Augustin,J.-C. and Carlier,V. (2000) Modelling the growth of Listeria monocytogenes with a multiplicative
type model including interactions between environmental factors. International Journal of Food Microbiology
56, 53-70.
Baranyi, J. and Roberts, T.A. (1994). A dynamic approach to predicting bacterial growth in food. International
Journal of Food Microbiology 23, 277-294.
Baranyi, J., Pin, C. and Ross,T. (1999) Validating and comparing predictive models. International Journal of
Food Microbiology 48, 159-166.
Dalgaard,P. and Jorgensen,L.V. (1998) Predicted and observed growth of Listeria monocytogenes in seafood
challenge tests and in naturally contaminated cold smoked salmon. International Journal of Food Microbiology
40, 105-115.
Giménez,B.C. and Dalgaard,P. (2004) Modelling and predicting the simultaneous growth of Listeria
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