Factors determining survival and

growth of micro-organisms in
foods

Frédéric Carlin
UMR 408 Sécurité et Qualité des Produits d’Origine
Végétale

https://www6.paca.inrae.fr/sqpov

Centre PACA Université d'Avignon, UFR-ip STS

Domaine Saint-Paul Campus Jean-Henri Fabre, Pôle Agrosciences
Site Agroparc
CS 40509 301, rue Baruch de Spinoza, BP 21239
84914 Avignon Cedex 9 84911 Avignon Cedex 9.

Titre de la présentation p. 1
Date / information / nom de l’auteur
 Combien de victimes de toxi‐
infections alimentaires en France ? 
500 décès*
(Estimation InVS – SantéPublique France)

2000

8000

*Pour > 100 milliards de portions d’aliments
 Incidence
Cas Cas décès
hospitalisés
Bactéries 58000- 11000- 200-700
91000 20000
Virus 500000 1000-1500 32

Parasites 168000 1000 75

Estimations INVS, 2004

 Major causes of bacterial foodborne
poisonings (Estimations)
Hazard Number of cases/year (incidence/105) 

France, 90s USA 00‐08 Canada 00‐10

Bacillus cereus 460 (0.8) 63000 (21) 36000 (112)
Campylobacter 15000 (25) 840000 (280) 145000 (450)
C. botulinum 22 (0.04) 55 (0.02) 14 (0.04)
(foodborne)
C. perfringens 5900 (10) 970000 (323) 175000 (550)
E. coli STEC 560 (1) 175000 (59) 33300 (103)
L. monocytogenes 300 (0.5) 1600 (0.5) 180 (0.55)
Salmonella 36000 (61) 1000000 (345) 88000 (270)
S. aureus 6800 (12) 241000 (81) 25000 (77)
Détail des foyers de Tiac déclarés aux ARS ou 
DDPP, France, 2012 – Agents confirmés
Burden of disease, NL, 2009
Hazard DALY (Year/100000  DALY (year/1000  Severity
inhabitants) cases of 
illnesses)
Bacillus cereus 0.7 2.3 1
Campylobacter 19.8 41 2
C. botulinum _ _ 3
C. perfringens 3.2 1
L. monocytogenes 0.7 1450
Salmonella 7.7 49 2
S. aureus 4.7 2.6 1
Virus Hepatitis A 0.9 167 3
Gastroenteritis viruses 8.9 2.4 1
Cryptosporidium 0.4 2.9 1
T. gondii (congenital) 13.8 6360 4

DALY Disability Adjusted Life Years.

.07
Havelaar et al. IJFM 2012
….il se confirme qu’un innocent croupit dans la prison de Sétif depuis 1998. Il y a été
incarcéré à l’issue de parodies de justice, des procès au pas de charge….
Il s’agit de Monsieur CHOUAR Bouguerra, condamné lors d’un premier procès, en
1998, à la peine capitale. En 2003.
Cette peine a été commuée en détention à perpétuité en 2003.
Puis lors de deux autres procès qui se déroulèrent en 2006, cet homme fut condamné à
deux peines de prison de dix années de détention chacune.
Ces procès à répétition, dont de nombreux experts diront qu’ils ont été conduits en
violation flagrante des procédures légales en vigueur et des droits les plus élémentaires
de l’accusé, se sont fourvoyés dans des considérations de toutes sortes, dont celle de
détourner les effets d’une juste vindicte populaire sur un bouc émissaire désigné pour la
circonstance.
Cette affaire, dite du cachir avarié, remonte à l’été 1998.
Des cas de botulisme s’étaient déclarés dans plusieurs localités de l’Est algérien, et
avaient causé la mort de 44 personnes, ainsi que l’hospitalisation d’une centaine
d’autres. Cette affaire qui avait été abondamment relayée par la presse, et tout
particulièrement par la télévision, avait suscité une immense clameur.

Décembre 2009 http://lequotidienalgerie.org/2009/12/20/un-innocent-

croupit-en-prison-depuis-1998/
Generalities

Titre de la présentation p. 12
Date / information / nom de l’auteur
 Different types of factors (1)
Intrinsic factors pH, aw, redox
- Food specific, Nutrients, preservatives
- Direct-contact with
micro-organisms

Extrinsic factors Temperature, gas

- External application to the atmosphere
food Relative humidity
- Efficiency during storage

13
 Different types of factors (2)
Implicit factors
- Specific to micro- – Maximal specific growth
organisms rate
- Interactions - Synergism and antagonism

Factors associated to Cutting, washing, packaging,

processing irradiation, pasteurisation

14
D’après Gould 2000
How to determine the effect of a factor?

• Estimation of changes with time in the number

of viable cells in a population
• Qualitative estimation
- Growth/no growth
- Turbidity in growth media
• Quantitative estimation
- Enumeration (formation of colonies) along time
- Following absorbance

15
Growth / no growth interface
E. coli

Slater et al. 2000 16

Bacterial growth and
parameters
10

8 Ymax

7
µ
Y

Yo 4 λ : lag
3
0 100 200 300 400 500 600 700

Time
17
 Survival curve
10
Log10 UFC/ml ou/g

0
0 2 4 6 8 10 12

TIME

18
Staphylococcus aureus survival curves

HEAT (58°C) PEF (26 kV cm-1)

HP 450 MPa

Cebrian et al., JAM, 2009

19
Target in Micro-organisms for
inactivation

20
Sensitivity to osmotic stress

21
Clark and Ordal AM 1969
 Cell damages created by physical and chemical 
inactivation treatments (Mackey 2000)
Symptoms of damages Possible cause

Sensitivity to low pHs Membrane damages 
Osmotic stress
Sensitivity to bile salts, antibiotics,  External Membranes Gram‐,  Damages to surface 
lytic enzymes proteins for Gram+
Sensitivity to oxidative stress Catalase, SOD inactivation. Increase in H2O2 and O2‐
(ROS). Loss in cellular reductors, cofactors of DNA 
repair enzymes

Changes in nutrient requirements Enzymatic degradation.
Damages to membranes leading to changes in 
accessibility enzyme – substrate
Increase in mutation rate DNA Damages

Extension of lag Need for synthesis of new membranes, DNA, RNA, 
ribosomes, etc…

22
Effect of a physical treatment
‐ A case study. Inactivation by heat

Titre de la présentation p. 23
Date / information / nom de l’auteur
Heat treatment
• Processing foods to make them 
edible
• Enzymatic and Microbiological 
stability
• Heat = energy
• Energy = f (time, temperature) 
• Inactivation of microorganisms 
= supplying enough energy to 
inactivate “ targets ”

24
 Heat-resistance parameter (1)

9
log10 cfu/g or /ml
8
Dt°C = 5 min
7

5
D
4

3
0 5 10 15 20
TIME at a given temperature (min)
25
Heat-resistance parameters (2)
100

z = 15 °C
D (min) 10

1
z

0.1
70 80 90 100

TEMPERATURE (°C)

26
 Mathematical models

Primary model

Secondary model

27
Heat-resistance of some non spore-
forming bacteria
Temperature Bacteria D Z (°C)
(°C) (min)
65 Thermophilic 100

Fecal streptococci 5 – 30

Lactobacillus spp. 0.5 – 1 8 - 10

Leucosnotoc spp. 0.5 – 1 8 - 10

L. monocytogenes 0.3 – 1

E. coli 0.1

55 P. fluorescens 1-4
28
Heat-resistance of some spore-
forming micro-organisms
Temperature Bacteria D (min) Z (°C)
(°C)
121.2 M. thermoacetica 20-90

G. stearothermophilus 4-5 8 -12

C. botulinum Groupe I 0.1 – 0.2 8 - 10

C. sporogenes 0.1 – 0.15 8 - 10

B. coagulans 0.01 –0.1 8 - 10

100 B. cereus 5 10

C. perfringens 0.3 – 20

B. polymyxa 0.1 – 0.5 7-9

29
 Heat resistance / Yeasts
Microorganism Broth or medium Temp D
(°C) (min)
Debaromyces hansenii YMPG, pH 3.5 48 10
Rhodotorola rubra YMPG, pH 3.5 50 26
Saccharomyces uvarum Citrate-P buffer 55 0.07
Saccharomyces uvarum Citrate-P buffer 55 8

Saccharomyces cerevisiae Apple juice, pH 3.6 55 0.9

Saccharomyces cerevisiae, Apple juice, pH 3.6 55 106
ascospores
Saccharomyces cerevisiae, Apple juice, pH 3.6 60 6.1
ascospores
30
 Heat resistance / Moulds
Microorganism Food or medium Temp D
(°C) (min)
A. Flavus – A. paraisticus conidia 60 0.1- 1

Neosartorya fischeri (ascospores) Cherry purée 85 52

Neosartorya fischeri (ascospores) Apple juice 88 1.4

Byssochlamys fulva (ascospores) Glucose- tartrate 90 17
Byssochlamys fulva (ascospores) Grapewine juice 85 56

31
Heat resistance / Viruses

EFSA 2015 32
 Sterilisation in canning
• F0, Heat-treatment at 121,2°C
• For C. botulinum, D121,2°C = 0,21 min et z
= 10°C
• For foods pH > 4.5-4.6, objective = 12D
sterilization process
• F0 = 12 x 0.21= 2.52 min ± 3 min

33
 Time temperature curve for can processing

http://www.nzifst.org.nz/unitoperations/httrapps2.htm

34
Calculation of F values. Summation of intervals

35
 Pastereurizing value (1)
• Definition : time à 70°C allowing the
same inactivation as the one obtained in
the applied heat process.
• Reference bacterium = Streptococcus
faecalis
• D70°C = 3 min, z = 10°C

36
 Pasteurizing value (2)
• For a treatment at constant temperature and
for time t

• 1 min at 70°C, VP = 1
1 min at 90°C, VP = 100
20 min at 80°C, VP = 200
• For a treatment at a changing temperature,
with T = f(t)
37
 Factors modulating heat-
resistance
• Intraspecies variability
• Before treatment. Physiology
- How tested cells are formed
- Stress before stress
- Growth stage
• During treatment. Composition of the heating
matrix
- pH, aw, fat content
• After treatment. Recovery conditions
38
 Variability of B. cereus heat‐resistance

1000

100
log D (min)

10

0.1

0.01
75 85 95 105 115 125

TEMPERATURE

39
From 19 papers
Genetics and heat‐resistance

Berendsen et al. ISME J 2015

 Temperature shock and
resistance to heat

B. subtilis PS346 CONTROL

30°C Log10 D (min)
HS = 30 min to 10
48°C Heat-shock
CS = 30 min at
10°C
Cold
Schock
1
80 85 90 95 100 105
Temperature °C
Movahedi and Waites JB, 2000, 2002
Growth stage
and heat
resistance

Lou and Yousef JFP 1996

 aw and Salmonella heat-resistance

Milieu pH (aw) Solute °C D (min)

P buffer, pH 6.5 6.5 (nc) None 65 0.3

P buffer, pH 6.5 6.5 (nc) Sucrose or Glucose 30:70 (% 65 1 – 43

w/vol)

Heart broth 7.4 (0.99) None 60 6.1

Heart broth 7.4 (0.90) NaCl 60 2.7

Heart broth 7.4 (0.90) Sucrose 60 75.2

Milk chocolate (98 % DM) nc nc 70 440

(Farkas, 1997) 43
 Fats and B. cereus heat-resistance

Matrix Temperature D (min) z (°C)

Buffer 95.6 °C 16 12.2

Buffer 109 °C 1.3

Olive oil 110°C 49.3 27

Olive oil
128°C 9.4

44
(Ababouch et Busta, 1987)
Fats and C. botulinum heat-resistance

Matrix Temperature D (min) z (°C)

Buffer 100°C 16 11.2

Buffer 114°C 1.3

Olive oil 109.5°C 25.8 28.5

Olive oil
129.2°C 5.3

45
(Ababouch et Busta, 1987)
pH and Bacillus coagulans heat-resistance

Palop et al., 1999

46
Recovery and incubation
temperature

47
MacKey et al. LAM 1994
Oxygen and growth recovery after heat-treatment (1)

● 10 % H2
○ 100 % N2
▲ 0,5 % O2
△ 1 % O2
◆ 2 % O2
◊ 10 % O2
■ 20 % O2
□ 40 % O2
 Reactive oxygen species and
detoxifying system
Respiration =
- energy production
- with ROS generation

ROS
Anion superoxide O2−,
Hydroxyle radical (·OH).
Hydrogen peroxide H2O2
Singulet oxygène O2•

https://fr.wikipedia.org/wiki/D%C3%A9riv%C3%A9_r%C3%A9actif_de_l'oxyg%C3%A8ne#/media/File: 49
Systeme_detox_ROS.GIF
Oxygen and growth recovery after heat-treatment (2)

Gas atmosphere Eh (mV) E. Coli O157:H7

6D at 59°C
Anaerobiosis from -350 to –50
19 – 24 min
0,5 – 1 % O2 from + 50 to + 100 5 – 17 min

2 – 40 % O2 from +100 to +380 3 min

George et al., 1998 50

Heat-resistance and germination of non
proteolytic C. botulinum spores

Survivor counts (log ufc/ml) + Lysozyme

Time at 85°C (min)

Peck et al. 51
1992
Effects of cooking on foods –
Comparison with effects on micro‐
organisms 

Titre de la présentation p. 52
Date / information / nom de l’auteur
 Cooking vegetables
Vegetable Temperature Cooking time

Potato 110 °C 5 min

100°C 10 min

90°C 40 min

Root vegetable 100°C 4 – 13 min

53
Heat-treatment and enzymatic
stability
Temperature D peroxydase Peroxydase
inactivation time

82°C 5 min 15 min

90°C 1.05 min 3.2 min
98°C 0.4 min 1.2 min

54
 Thermal resistance of aerobic spore formers isolated from
food products

International Journal of Dairy Technology

Volume 64, Issue 4, pages 486-493, 22 JUL 2011 DOI: 10.1111/j.1471-0307.2011.00706.x
http://onlinelibrary.wiley.com/doi/10.1111/j.1471-0307.2011.00706.x/full#f3
 Other physical inactivation
treatments
• Irradiation (KGy)
• High Pressure (Pascalisation) (MPa)
• Micro-waves
• Pulsed Electric Fields
• Magnetic fields
• UV treatments (J/m2)
• Pulsed UV-light
• …

56
 Non predictible effects (1)

• Heat
- viruses, bacteria, yeast < molds < bacterial
spores
• UV
- Bacteria, spores of bacteria < molds
• Irradiation
- Bacteria < spores << viruses

57
 Non predictible effects (2)

Strains D 100 °C (m in) D 588 Mpa (min)

B. stearothermophilus 1 339 ∞

B. stearothermophilus 2 46.8 91

B. coagulans 4.79 Activation

B. subtilis 0.85 ∞

B. licheniformis 1.02 ∞

B. megaterium 0.19 313

58
Nakayama et al. 1996
 Light Vs Heat (bacterial spores)
1.2 A.acidoterrestris
Gb.stearothermophilus

1
B.megaterium
B.cereus KBAB4
0.8
F3 value (J/cm ²)

0.6 B.subtilis B.pumilus

0.4
B.licheniformis
0.2

0
0 10 20 30 40 50 60 70 80 90 100
t3 value (min)

• No correlation between the t3 value and the F3 value

• The most heat resistant species are not necessarily the most 
Pulsed light resistant
Levy et al., IJFM, 2012
Controlling growth

Titre de la présentation p. 60
Date / information / nom de l’auteur
 Factors determining microbial growth

• pH
• Water activity
• Redox potential
• Incubation temperature
• Preservatives
• ….

61
How to determine the effect of a factor?

• Estimation of changes with time in the number

of viable cells in a population
• Qualitative estimation
- Growth/no growth
- Turbidity in growth media
• Quantitative estimation
- Enumeration (formation of colonies) along time
- Following absorbance

62
Bacterial growth and
parameters
10

8 Ymax

7
µ
Y

Yo 4 λ : lag
3
0 100 200 300 400 500 600 700

Time
63
 pH of some foods
Food pH
Fermented shark 11
Hen egg white 9
Fish 6.5 – 7.5
Milk 7
Meat 5.5 – 6.5
Wheat flour 6–7
Vegetables 4.5 – 7
Fruits 2 – 4.5 64
 pH and growth rate (1)
µ

min pH Opt pH Max pH

pH 65
pH and growth rate (2)

66
Rosso et al., AEM, 1996
pH et pKa

67
pH et pKa

68
 Physiological effects

Plasma membrane

pH, neutral

HA A- + H+
ATP

ADP

HA A- + H+ 69
 pH growth limits for bacterial
foodborne pathogens
Bacteria min pH, opt pH
L. monocytogenes 4.4, 7
B. cereus 5.0, 6.0-7.0
C. botulinum 4.6, 7.0
Vibrio spp. 4.8, 7.8
Salmonella 3.8, 7-7.5
E. coli, including O157 :H7 4.4, 6-7
S. sonnei 4.9, 7.0 ?
S. aureus 4, 6-7
C. perfingens 5.5-5.8, 7.2
Campylobacter 4.9, 7.0

70
How to determine water activity?

Pure water
Equilibrium vapor pressure = P
Equilibrium vapor pressure = P0

aw = 1 aw < 1
P1 Hygrometry1
aw  
P0 Hygrometry0
0 < aw < 1 71
 Osmoregulation
High aw of external medium Low aw of external medium
(> cell) (< cell)

H2O H2O
- Inner osmotic pressure increase
- Swelling - Accumulation of K+, glutamate
- Mechanical resistance of cell - Compatible solutes
wall 72
Growth of salt stressed L. monocytogenes in
the presence of glycine - betaine or carnitine

Avg Doubling time (h) ± SD

LM1 LM2 LM3

No addition NG 44 ± 5 NG
100 µM GB 16 ± 0.6 11 ± 0.1 16±0.2
100 µM CAR 94 ± 2 13 ± 0.2 NG
5 mM CAR 42 ± 2 12 ± 0.6 81 ± 3

Mendum and Smith AEM 2002 73

 aw of some foods

1,00 – 0,95 M eat, fish, fresh m ilk, fresh cheese

0,95 – 0,90 Cooked ham , salam i, bacon, hard chesse
0,90 – 0,85 M atured hard cheese, fruit juice concentrate, M arple syrup

0,85 – 0,80 Milk concentrate with sugar, parm eggiano chesse, fruit
cakes
0,80 – 0,75 Salted fish, jam and m armelade, prunes

0,75 – 0,65 Dates, figs, sultanas, walnut

0,65 – 0,60 Dry fruits, honey

< 0,60 Dry pasta, spices, biscuits, m ilk powder, dry legum es, flour,
cereals, instant coffee
74
 Composition of some foods in
relation to their aw
• Raw cow’s milk (1 L)
- water 900 g, fat 35-40 g, proteins 30-35 g, lactose 50
g, minerals 10 g (total ± 130 g)
• Raw meat: 70 % water
• Raw fish: 80 %
• Cooked ham : 70 % water, 2 % salt
• Raw ham: 3.5 % salt, 50 % water
• « saucisson »: 35% water. Salt 5 %
• Jam: 50 % water – 50 % sugar

75
NaCl and sugar concentrations,
and aw

76
Christian 2000
 Fruits

Pectin
Starch
Wheat
Meat, fish
Cellulose

Water sorption isotherm at 20°C (ICMSF 1980)

77
Growth rate and aw

78
Leroi el al. FM 2012
Minimal aw for growth of some
bacteria
Bacillus cereus 0.93
Clostridium botulinum 0.94
Clostridium perfringens 0.97
E scherichia coli 0.95
Listeria monocytogenes 0.92
Salmonella species 0.95
Staphylococcus aureus 0.86
Halobacterium halobium 0.75

79
« Hazelnut yoghurt », UK, 1989 (1)

M. Peck, IFR
N i h
« Hazelnut yoghurt », UK, 1989 (2)

M. Peck, IFR
N i h
Minimal aw for growth of some
yeast and molds
Aspergillus flavus/ parasiticus 0.80
Botrytis cinerea 0.97
Eurotium spp 0.62 – 0.74
Penicillium spp. 0.79 – 0.82
Rhizopus stolonifer 0.89
Xeromyces bisporus 0.61
Saccharomyces cerevisiae 0.90
Zygosaccharomyces rouxii 0.62
82
 Oxidation –reduction in foods
• [oxydant] + H+ + ne- ⇋ [Reductant]
• With respect to Oxygen Transfer
- Oxidation is the gain of oxygen
- Reduction is the loss of oxygen
• With respect to Hydrogen Transfer
- Oxidation is the loss of hydrogen.
- Reduction is the gain of hydrogen.
• Oxidizing agents add oxygen to another substance or remove
hydrogen from it.
• Reducing agents remove oxygen from another substance or add
hydrogen to it.
• With respect to Electron Transfer
- Oxidation is loss of electrons (OIL)
- Reduction is gain of Electrons (RIG)
• http://chemwiki.ucdavis.edu/Analytical_Chemistry/Electrochemistry/Definitions_of_Oxidation_and_
Reduction 83
 Redox potential
• [oxydant] + H+ + ne ⇋ [Reductant]
• Nernst Equation
Eh = E’O + RT/nF Log[oxydant] [H+]/[Reductant]
- E0 = Reference redox potential
- R = perfect gases constant
- Number of electrons implicated in the reaction
- F = Faraday constant

84
Ascorbate, a reducing agent

Ascorbic acid Dehydroascorbic acid

C6H8O6 + ½ O2 ⇋ C6H6O6 + H2O

http://en.wikipedia.org/wiki/Vitamin_C#mediaviewer/File:Ascorbic_acid_structure.png 85
 Some redox couples in foods
Couple redox E0 (mV)
½ O2 /H20 +820
Fe3+/Fe2+ + 720
Cytochrome C (ox/red) +250
- Dihydroascorbic/Ascorbic acid +80

Pyruvate/lactate -190
Glutathione oxid./Glutathion red. - 230

NAD+/NaDH -320

86
 Redox potentiel of some foods
Aliment pH Eh (mV)
Raw meat 5,7 - 200
Raw minced meat 5,9 + 225
Wheat (whole grain) 6,0 -320 to –360
Barley (ground grain) 7,0 + 225
Cooked meats 6,5 -20 to –150
Cooked dishes meat chicken 5,5-6,1 -20 to –200
Potato tuber 6,0 -150
Lasagna 4,7 -23
Spinach 6,2 + 74
Pear 4,2 +436
Grape 3,9 + 409
Lemon 2,2 +383
87
Snyder 1996, Adans et Moss 1995)
 Factors influencing redox potential Eh
in foods
• Nature of redox couples
• Ratio of oxidant to reductant
• pH
• Poising capacity (redox buffer)
• Availibility of oxygen
• Microbial activity

88
 Limited oxygen availibility in foods
• Vacuum‐packaging
• Modified oxygen
• Oxygen absorbers
• Reduction by oxidation of food components
• Reduction by the activity of the microflora
• Still too much oxygen for anaerobes?
• Not enough for facultatively anaerobes?
90
91
 Respiratory status of micro-organisms

Aerobic/
Energy Redox Toxicity
Type Anaerobic Examples
yield domain of O2
respiration

+100 –
Strict aerobes +/- +++ - Ps. fluorescens
500 mV

Facultative
anaerobes +/+ +++/+ ? - Enterobacteriaceae

Facultative
(aerotolerant) -/+ -
-/+ ? - Lactic acid bacteria
aerobes ++

Strict -/+ -
-/+ ? + Clostridium sp.
anaerobes ++

92
Redox potential, O2, and growth of
C. botulinum
Eh (mV) P germination
O (%)
2
0 - 400 1

0,21 + 180 0,28

0,39 + 210 0,23

0,72 + 245 0,11

0,79 + 250 1 x 10-4

1,12 + 271 1 x 10-5

1,30 +280 2 x 10-5

1,67 + 300 4 x 10-6

93
Lund 1993
Solubility of O2
Oxygen uptake by the microflora

Robinson LAM 1994
 Consequences of anaerobiosis

• Decrease in growth rate
• Decrease in ATP production
• Different metabolic pathways
• Use of electron acceptors other than oxygen
 Metabolism changes in B. cereus

µmax  Biomass % glucose  Yield (mol.mol‐1)

(h‐1) (g.L‐1) used

Acetate lactate Formate Ethanol

MOD +  1.65 2.7 90 0.95 < 0.01 < 0.01 < 0.01
glucose, 
aerobiosis
MOD +  0.95 0.8 100 0.22 1.37 0.30 0.05
glucose 
anaerobiosis

Rosenfeld et al. CJM 2005 
 B. weihenstephanensis. 
Potato purée
Parameter Initial O2 % in headspace
20.9 10 5 1 Vacuum
Lag (days) 5.7 5.9 5.8 6.5 9.1
µmax (h‐1) 1.5 0.68 0.74 0.78 0.81
Nmax 7.7 6.2 6.11 5.4 4.5
105 B. cereus cfu/g à  7.2 9.8 9.3 10.8 14.4
(days) : 

Samapundo et al., FM, 2011
Effect of temperature on growth
µ

Tmin Topt Tmax

Temperature
99
Effect of temperature on Listeria
monocytogenes growth
L m ono: effet T °

1. 4

1. 2
µ (h-1)
1

0. 8
µ(1/h)

0. 6

0. 4

0. 2

0
0 5 10 15 20 25 30 35 40 45 50

F
T e m p érat u re (°C )

S O R1 00 S O R 10 1 UNIR 10 0 UNI R 101 UNI R10 2

Temperature
INR A 100 I NRA 1 01 A TC C1 911 5 S O R 10 0 L m o no (I nra)
µ(h-1 ) b orne_ inf bo rne_s up

100
 Cardinal temperatures for
growth
Tmin Topt Tmax
(°C) (°C) (°C)
Thermophiles 40-45 55-75 60-90

Mesophiles 5-15 30-40 40-47

Psychrophiles -5 - +5 12-15 15-20

Psychrotrophes -5 - +5 25-30 30-35

101
Cardinal temperatures for growth of
pathogenic bacteria
Tmin (°C) Topt Tmax (°C)
(°C)
B. cereus* 4-15 28-35 40-55
Campylobacter 32 42-45 45
C. botulinum Gp I 10 35 50
C. botulinum Gp II 3 30 45
C. perfringens 12 43-47 50
E. coli 7 35-40 46
L. monocytogenes 0-4 30-37 45
Salmonella 5 35-37 45-47
S. aureus** 7 35-40 48 102
Growth during cooking and
cooling

Perfringens predictor at www.combase.cc

Le Marc et al.
 Physiological effects
• Low temperature
- Keeping exchanges
- maintaining plasma membrane fluidity
- Conversion of fatty acids (saturated
→unsaturated)
- Decrease of enzymatic activity
• High temperature
- Protein denaturation
- Alteration of plasma membrane
104
Quelle température réelle ?

Derens-Bertheau
IJR 2015
 Lois empiriques
• Modèles empiriques (Ratkowsky)
- õ = b (T РTmin)
õ

Tmin

106
 Facteur Température
 max   opt . T 0 < T < 1
(T  Tmin ) 2 (T  Tmax )
T 
(Topt  Tmin )(Topt  T )(T  Topt )  (Topt  Tmax )(Topt  Tmin  2T )
min

µopt

Tmin Topt Tmax

Taux de croissance de E. coli en fonction de la température

107
 Preservatives
Code Preservative
E200-203 Sorbic acid and derivates
E210-218 Benzoic acid and derivates
E220-227 Sulfites
E234 Nisin
E235 Pimaricine (natamycine)
E240 Formaldehyde
E249-252 Nitrites and nitrates
E260-263 Acetic acid and derivates
E270 Lactic acid
E280-282 Propionic acid and derivates
E284-285 Boric acid and derivates 108
 Sorbic acid
• GRAS
• Low toxicity (acceptable daily intake > NaCl !)
• Large application in foods
• CH3-CH=CH-CH=CH-COOH
• Higher solubility of salts (K)
• Large spectrum of action, in particular yeasts
and molds
• Applications : fermented milk, yoghourts,
confectionery, cakes, prunes, candied fruits,
margarines, cereals 109
110
111
 C. botulinum and sorbic acid
pH Total Sorbic acid (mg/l) - Log10 P à 30°C after
3d 14 d
7.0 0 0 0
2000 -0.12 -0.12
5.5 0 0.15 0.15
1000 3.56 0.28
1800 7.65 5.65
4.9 0 1.2 1.2
370 > 6.1 1.5
670 > 8.1 8.1> 112
Lund et al. 1987
 Properties of sorbic acid
• Weak acid
• pKa = 4.76
• pH < 4.76 : mainly AcH
• pH > 4.76 : mainly Ac-
• Only AcH has antimicrobial activity

113
 C. botulinum and sorbic acid
pH Ac. Sorbique (mg/l) - Log10 P à 30°C après
Total Non dissocié 3j 14 j
7,0 0 0 0 0
2000 11 -0,12 -0,12
5,5 0 0 0,15 0,15
1000 156 3,56 0,28
1800 281 7,65 5,65
4,9 0 0 1,2 1,2
370 157 > 6,1 1,5
670 283 > 8,1 8,1>
114
Lund et al. 1987
Interactions between factors

www.foodprocessing-technology.com 115
 Interactions and growth of
C. botulinum group II
pH NaCl (%) C. botulinum, types B, E, F
Time to growth at the indicated temperature:

16°C 12°C 5°C

6.5 0.6 2 2-3 12

6.5 2.5 2 4 24-27

5.6 0.6 2 4 10-18

5.6 2.5 5 6 31-47

6.5 + 85°C-18 min 0.6 55 - > 104 76 - > 104 > 104

5.6 + 85°C-18 min 2.5 > 91 > 91 >116

91
Graham et al. 1996
A model forC. botulinum

117
M. Peck, IFR Norwich
 Objectives of predictive
microbiology
• “Microbiology” and “prediction”
• “Predictive” and “modelling”
• Prediction of survival and growth
• Food industry
• Food safety and stability
• Factors relevant to the food industry
• However a relevant question for biology
 Why predicting? Why models?
• Some evidences
- The more I cook, the more I kill
- Some temperatures are better than others for growth
- Biological limits for growth
- High acidity or high salt do not help….
• And ??????
- How much more rapid growth at 20°C than at 10°C?
- Less salt for healthier foods, fine! But consequences on safety?
What is acceptable?
- Safe shelf-life of foods: how long?
• Quantitative data linked with mathematical equations
- Predicting (mathematical) models
 Lois empiriques
• Modèles empiriques (Ratkowsky)
- õ = b (T РTmin)
õ

Tmin

120
 Facteur Température
 max   opt . T 0 < T < 1
(T  Tmin ) 2 (T  Tmax )
T 
(Topt  Tmin )(Topt  T )(T  Topt )  (Topt  Tmax )(Topt  Tmin  2T )
min

µopt

Tmin Topt Tmax

Taux de croissance de E. coli en fonction de la température

121
 Factor pH
 max   opt . pH 0 < pH < 1

  pH  
 pH  pH  pH  pH 
min max

 pH  pH  pH  pH    pH  pH 
min max opt
2

Effect of pH on the growth rate of L.innocua

 Facteur activité de l’eau (aw)
 max   opt . aw 0 < aw < 1

 aw 
aw  aw  aw  1 min
2

aw
opt
 awopt  awmin aw  awopt   awopt  1awopt  awmin  2.aw 
 awmin 

0.5

0.45

0.4

0.35

0.3

0.25

0.2

0.15

0.1
0.8 0.82 0.84 0.86 0.88 0.9 0.92 0.94 0.96 0.98 1

123
 Factor organic acid
 max   opt . AH   AH  


  AH   1   
 CMI 

AH 
acide
1  10 pH  pKa

CMI = 9,5 mM
= 0,71

Growth of L. monocytogenes as function of Acetic acid

concentration pH=5.1 and pH=5.4
 Modular approach
« Hurdle technology »

 Gamma Concept of Zwietering (1992)

• Multiplicative effect of factors

• Independant interactions

µmax = µopt (T) (aw) (pH) g(AH)

g(AH) g(interaction)

4 environmental factors

Specific matrix effect