Microbial spoilage of liquid egg products and its control by the detection of genetic or chemical markers. journée scientifique "science des aliments et valorisation des bio-produits

Jan, Sophie

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Microbial spoilage of liquid egg products and its control
by the detection of genetic or chemical markers
Sophie Jan
To cite this version:
Sophie Jan. Microbial spoilage of liquid egg products and its control by the detection of genetic or
chemical markers. journée scientifique ”science des aliments et valorisation des bio-produits”, Institut
National de Recherche Agronomique (INRA). UMR UMR INRA / AgroCampus Rennes : Science et
Technologie du Lait et de l’?uf (1253)., Apr 2019, Rennes, France. ￿hal-02114761￿
Prénom Nom - Equipe
STLOpen Days
19-21 March 2019
Microbial spoilage of liquid egg products and its control 
by the detection of genetic or chemical markers
Sophie JAN
MICROBIO Team
STLOpen Days
19-21 March 2019
2
One of the leader in the production and consumption of eggs
in the EU
 14.9 billion eggs produced / year
 213 eggs consumed /year / person 60 – 70 % 30 – 40 %
Itavi and AND International, 2017
Introduction
Table  eggs Egg products
Biscuits, pastries,  
desserts, pasta, chocolate 
and sugar confectionery, 
delicatessen, seafood 
products, wines, etc.
Foaming
Emulsifying
Coloring
Binding
Thickening
Flavoring
Source of ingredients
Source of nutrients
Food artisans
Industrial catering and residential facilities
Hospitals
Restaurants
How to control microbial spoilage in the sector of egg processing ?
A key challenge for the reduction of food wastes
Food industry
Egg products
STLOpen Days
19-21 March 2019
3
What about egg microbiology?
Eggshell surface
Contaminated in the environments of hen breeding and egg handling
and storage
+++ Gram-positive bacteria
Streptococcus, Aerococcus, Micrococcus, Staphylococcus, and low
occurrence of Bacillus
+  Gram-negative bacteria
Escherichia coli, Yersinia, Enterobacter, Klebsiella, Serratia,
Citrobacter, Salmonella, Alcaligenes, Pseudomonas
Nead for egg product stabilization for ensuring egg product safety
Sterile except under sporadic cases of
contamination by Salmonella (mainly
Salmonella Enteritidis) during egg
formation
Less than 1% of contamination but still 30% of 
strong evidence of salmonellosis caused by eggs or 
egg products in the EU
• Strong affinity for the avian reproductive tract
• Persistence on the eggshell surface
• Resistance to egg defence systems
Egg content
EFSA and ECDC, 2017
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19-21 March 2019
4
What are the main industrial processes ensuring egg product safety?
Filtration
Pasteurisation
Packaging
Storage
Delivery
Mild treatmentsRaw egg products
Destruction of heat sensitive bacteria, 
including Salmonella
Eggshell breaking
Separation of the 
egg compartments
Shell egg
Cooling
T° ≤ 4°C
How to control heat-resistant psychrotrophic bacteria in liquid egg products 
and sensitive food where they are used as ingredients?
What about  heat-resistant
bacteria?
What about 
psychrotrophic
bacteria?
STLOpen Days
19-21 March 2019
5
What about the microbiology of egg-based sensitive food?
What are the 
microorganisms coming 
from egg products ?
How to avoid their presence 
and/or to control their 
development ?
?
Demand for the development of industrial efficient tools to detect food 
spoilage contaminants or the products of their metabolism
Industrial egg-based refrigerated desserts
The famous “île flottante” dessert as a 
research model
Custard cream comprising whole
egg or egg yolk as ingredient
Sweet egg white foam
Food waste
Microbial
development
Food spoilage
Customer
complaints
STLOpen Days
19-21 March 2019
6
What are the main bacterial contaminants of liquid egg products?
Egg Yolk
Whole egg
Perfect media 
for bacterial 
growth
Heat-treated at 65 
to 68 °C for 5 to 6 
min in France 
What are the main 
contaminants of liquid
whole egg products?
Egg white
Poorly 
adapted to 
microbial 
growth
Heat-treated at 55 
to 57 °C for 2 to 5 
min in France
What are the main 
contaminants of liquid
egg white products?
STLOpen Days
19-21 March 2019
7
What are the main bacterial contaminants of liquid egg products?
Whole egg
Perfect 
medium for 
bacterial 
growth
Heat treatment
Main contaminants
Bacillus (including Bacillus cereus group)
Enterococcus
Streptococcus
Pseudomonas
Lactobacillus
Techer et al., 2014 ; Miller et al., 2010 ; Baron et al.,
2007 ; Protais et al., 2007 ; Protais et al., 2006 ;
Wood et Waites, 1988 ; Foegeding et Stanley, 1987 ;
Payne et Gooch, 1980 ; Shafi et al., 1970
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19-21 March 2019
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What are the main bacterial contaminants of liquid whole egg products?
Industrial finished
whole egg products
coming from 5 
French industries
Enrichment of the 
Gram positive flora
Mesophilic Bacillus cereus group bacteria
100 % of the companies
76 % of the samples
Psychrotrophic Bacillus cereus group bacteria
100 % of the companies
44 % of the samples
Higher occurrence at the cold season
37°C
10°C
Bacterial collection of 68 representative psychrotrophic B. cereus group 
bacteria
Koné et al., 2013 BPS
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19-21 March 2019
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What are the main features of the bacterial collection coming from whole egg products?
68 psychrotrophic B. cereus
group bacteria
Risk
ealth
Risk
conomic
Belonging to a phylogenetic
group (panC sequence) 
(Guinebretière et al., 2008)
Growth, biofilm formation
Expression of activities
Toxinogenesis
Spoilage
Genetic markers of the 
psychrotrophy
Jan et al., 2011. Food Microbiol.
STLOpen Days
19-21 March 2019
10
What are the main features of the bacterial collection coming from whole egg products?
68 psychrotrophic B. cereus
group bacteria
Belonging to a phylogenetic
group (panC sequence) 
(Guinebretière et al., 2008)
Jan et al., 2011. Food Microbiol.
A high diversity inside the 
collection
Five phylogenetic groups
Group VI 48.5 %
Group II 41.0 %
Group V 1.5 %
Group IV 7.5 %
Group III 1.5 %
STLOpen Days
19-21 March 2019
11
What are the main features of the bacterial collection coming from whole egg products?
68 psychrotrophic B. cereus
group bacteria
Risk
ealth
Risk
conomic
Growth, biofilm formation
Expression of activities
Toxinogenesis
Spoilage
Jan et al., 2011. Food Microbiol.
Spoilage as a function of time
Cytotoxic activity as a function of time
STLOpen Days
19-21 March 2019
12
What are the main features of the bacterial collection coming from whole egg products?
Expression of activities
Toxinogenic
Spoilage
Jan et al., 2011. Food Microbiol.
95 % of the collection
> 50 % of the collection
even at low temperature
at optimal temperature
Optimal growth
medium
Risk
ealth
Risk
conomic
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19-21 March 2019
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What are the main features of the bacterial collection coming from whole egg products?
-6 -4 -2 0 2
0
2
Principal component 1 (41.35 %)
P
rin
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p
a
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o
m
p
o
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1
0
.7
2
 %
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Pure psychrotrophs
High spoilage risk
4
Intermediate psychrotrophs
High health risk
An inverse correlation between
health and spoilage risks
A psychrotrophic signature in the 
cspA gene
4
group IV
group VI
group II
group III
group V
In red : isolates associated to health risk
In blue : isolates associated to spoilage risk
In garned red : isolates associated to high sanitary risk
LEGEND
Techer et al., 2014. J. Appl. Microbiol.
STLOpen Days
19-21 March 2019
14
What are the main features of the bacterial collection coming from whole egg products?
Jan et al., 2011. Food Microbiol.
An establised risk of 
biofilm formation
Biofilm formation
Strong adhesion
Stainless steel surfaces
STLOpen Days
19-21 March 2019
15
What are the main features of the bacterial collection coming from whole egg products?
Risk
conomic Spoilage
95 % of the collection
A genetic signature
What about the actual risks?
Risk
ealth
Only one isolate
 1.5 % of the collection
At low temperature
Selection by the process
An optimum medium
Industrial surfaces
An optimum environment
At optimal growth temperature
Expression of activities
toxinogenesis
> 50 % of the collection
No emetic strains
Isolates associated to high 
health risk
 10.5 % of the collection
Industrial cold
4°C
Techer et al., 2014. J. Appl. Microbiol.
STLOpen Days
19-21 March 2019
16
What are the main bacterial contaminants of liquid egg white products?
Egg white
Poorly 
adapted to 
microbial 
growth
Heat treatment
Main contaminants
No reference in the 
literature
STLOpen Days
19-21 March 2019
17
What are the main bacterial contaminants of liquid egg white products?
Egg white
Poorly 
adapted to 
microbial 
growth After heat-
treatment
Before heat-
treatment
100 isolates
66 isolates
Main contaminants
Techer et al., 2015, J. Food Science
STLOpen Days
19-21 March 2019
18
What are the main bacterial contaminants of liquid egg white products?
100 isolates
66 isolates
Bacillus (including Bacillus cereus group)
Enterococcus
Streptococcus
Pseudomonas
Lactobacillus
Enterobacter
Escherichia
Enterococcus
Serratia
Shigella
Klebsiella
Pantoea
Yersinia Rahnella
Salmonella
Pseudomonas
B cereus group
Fr
eq
ue
nc
y
Mainly coming from the eggshell surface contamination at the egg breading step
De Reu et al., 2005 and 2006 ; Musgrove et al., 2004 et 2005 ;
Mallet et al., 2006
Main contaminants
Techer et al., 2015, J. Food Science
Persistence after the heat-treatment
STLOpen Days
19-21 March 2019
19
What are the main bacterial contaminants of liquid egg white products?
100 isolates
66 isolates
Bacillus (including Bacillus cereus group)
Enterococcus
Streptococcus
Pseudomonas
Lactobacillus
Enterobacter
Escherichia
Enterococcus
Serratia
Shigella
Klebsiella
Pantoea
Yersinia Rahnella
Salmonella
Pseudomonas
B cereus group
Fr
eq
ue
nc
y
Enzymatic spoilage activities in 
custard cream even at low 
temperature
Main contaminants
Techer et al., 2015, J. Food Science
STLOpen Days
19-21 March 2019
20
Conclusion
Persistence of heat-resistant psychrotrophic bacteria in pasteurised liquid egg products
Spoilage activities, in the product itself and when used as ingredient, even at low temperature
Genetic signature of the spoilage risk due to psychrotrophic B. cereus group bacteria
And now?
Study of the bacterial spoilage metabolism
Development of online tools to early detect food spoilage contaminants or the products of their metabolism
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Microbial spoilage of liquid egg products and its control by the detection of genetic or chemical markers

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Microbial spoilage of liquid egg products and its control by the detection of genetic or chemical markers

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