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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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HAL Id: hal-02097231

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Submitted on 5 Jun 2020

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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. STLOpendays, Institut National de Recherche Agronomique (INRA). UMR UMR INRA / AgroCampus Rennes : Science et Technologie du Lait et de l’?uf (1253)., Mar 2019, Rennes, France. �hal-02097231�

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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

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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

(4)

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 occurrenceof 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

(5)

STLOpen Days

19-21 March 2019

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What are the main industrial processes ensuring egg product safety?

Filtration

Pasteurisation

Packaging Storage Delivery Mild treatments

Raw egg products

Destruction of heat sensitive bacteria, includingSalmonella

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?

(6)

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

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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?

(8)

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(includingBacillus cereusgroup) 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

(9)

STLOpen Days

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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STLOpen Days

19-21 March 2019

9

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 (

panCsequence)

(Guinebretière et al., 2008)

Growth, biofilm formation

Expression of activities Toxinogenesis

Spoilage

Genetic markers of the psychrotrophy

Jan et al., 2011. Food Microbiol.

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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 (

panCsequence)

(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 %

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STLOpen Days

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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

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STLOpen Days

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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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What are the main features of the bacterial collection coming from whole egg products?

-6 -4 -2 0 2

02

Principal component 1 (41.35 %)

Principal component 2 (10.72 %)

1 3 4 5

6 7 9 10

12 13

14 15 16

17 18

19

20

21 22

2423 25

27

28 29

30

31 32

33 34

35 36 37

38 39

40

41 42 43

44

45

46 47

48

49

50 51

52

53

54

55 56 57 58

59 60

61 62

63

64

65 66

67

68 8

26

2 11

Pure psychrotrophs High spoilage risk

4

Intermediate psychrotrophs High health risk

An inverse correlation between health and spoilage risks

A psychrotrophic signature in the cspAgene

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.

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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

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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.

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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

(18)

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

(19)

STLOpen Days

19-21 March 2019

18

What are the main bacterial contaminants of liquid egg white products?

100 isolates

66 isolates

Bacillus(includingBacillus cereusgroup) 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

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STLOpen Days

19-21 March 2019

19

What are the main bacterial contaminants of liquid egg white products?

100 isolates

66 isolates

Bacillus(includingBacillus cereusgroup) 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

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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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THANK YOU FOR YOUR ATTENTION

Please visit http://www.rennes.inra.fr/stlo_eng

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STLOpen Days

19-21 March 2019

21

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