Professional Documents
Culture Documents
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CRITERIA / STANDARD
(Codex alimentarius, CAC/GL 21 of 1997)
Micro-organisms of interest - interest for the product ; Sampling plan; Usable detection and/or quantification methods Satisfactory microbiological limits for the food at the critical point of the food chain ; The number of analytical units allowing a conformity compared to these limits ; The food to which it applies Stages of the food chain to which it applies ; All measures to be taken when it is not satisfied.
IAFP, Berlin, 2009
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Insects
Fungi Water
Environnement Protozoa Parasites
Toxicology ?
Example: Bacteria
Pathogen
1 microorganism
Bacteria resistant to antibiotics Opportunistic Deterioration flora Bioindicator flora Sanitary Technology Probiotics Floral with technological interest / Starter Flora of Ali-terrorism
IAFP, Berlin, 2009
1 Population, 1 ecology
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Serotype detection
Interesting public health: as EHEC Interesting veterinary sector
Time
PROCESS: ASSAY
Calibration
Methods ??
ANALYSES
Reagents Results Report
Quality Control
Measurement uncertainty
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Recording
Validation
Audit
Standardized methods
Reference text Elaborated by qualified body, with consensus among all the interested parties Continuous, repetitive and voluntary application Speak the same language
Distinction between full standard and Technical specification
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CEN standards
Scope: Standardization in the field of human and animal foodstuffs, covering the food chain from primary production to consumption, a well as animal and vegetable propagation materials, in particualr, but not limited to, terminology, sampling, methods of test and analysis, product specifications, food and feed safety and quality of management and requirements for packaging, storage and transportation. excluded: products covered by ISO/TC54 Essential oils and ISO/TC93 (Starch)
Functioning
ISO CASCO/BT ISO TC34
SC9
WG1
Meat and meat products
WG2 Statistics
WG4
Proficiency testing
WG5
Culture media
WG6
Parasites
WG7
Microbiological examinations
Project group
Probiotics
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Scope: Standardization of horizontal microbiological methods for all food and animal feeding stuff analysis and for any other sample that can be the source of microbiological contamination of food products
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Functioning
CEN BT
TC 327 Probiotics
TC 275
TC 302 MILK
ISO TC34/SC9
WG6 EU PROJECT
AdHoc group Ad(Hoc group C. sakazakii AdHoc group L. monocytogenes
Histamine, B. cereus toxins, Staph. toxins
TAG1
TAG2
TAG3 PCR
TAG4 Virus
TAG5
Primary
TAG6
TAG7
Media Validation
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Possible Introduction of the following text as a note in the scope of the standards on pathogen detection For epidemiological purposes or during outbreak investigations, other methods may need to be used in addition to this standard. A standard could not cover all matrices and all target microorganisms!
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Microorganisms
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Part 2: Enumeration
a new MPN method, able to analyze simultaneously a lot of samples at a lower cost. This technique is based on the ability of the MSRV medium to select and enriched motile Salmonella that may be further isolated on chromogenic medium
Part 3: Serotyping
Based on Kauffman whites scheme
IAFP, Berlin, 2009
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Campylobacter
(CEN TC275/WG6)
Revision of EN ISO 10272-1:2006 (detection Campylobacter) and of ISO/TS 10272-2:2006 (enumeration Campylobacter Colony-count technique). To improve the enrichment and confirmation steps and microaerobic incubation conditions
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Simplification of identification
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VTEC non-O157
(CEN TC275/WG6/adhoc group, S. Morabito (CRL)) o Project technical specification
o CEN ISO/WD TS 13136 Microbiology of food and animal feeding stuffs -- Horizontal method for the detection of Shiga toxin-producing Escherichia coli (STEC) belonging to O157, O111, O26, O103 and O145 serogroups -- Qualitative Real-time polymerase chain reaction (PCR)-based method
o First detection of stx,eae genes and after RT-PCR for serotypes followed by isolation of colonies o Problems of patents: RT-PCR for serogroup detection
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o A new need: An European standard for stx characterization by PCR or RT-PCR IAFP, Berlin, 2009
Enterobacter sakazakii
(CEN TC275/WG6/adhoc Group, H. Joosten & C. Iversen)
Draft: Horizontal method for the detection of Cronobacter spp. (Enterobacter sakazakii)
Relative poor performance with environmental samples Recovery some sensitive strains from some milk powders
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Brochothrix thermosphacta
(ISO TC34/SC9/WG2 (meat and meat products), E. De Boer)
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Gaz and acid production dacides from lactose (old definition) Escherichia coli
Escherichia coli Klebsiella Enterobacter Citrobacter Yersinia Serratia Hafnia In green, coliforms present in environment and human faeces
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Pantoea Kluyvera
Escherichia coli Klebsiella Enterobacter Citrobacter Yersinia Serratia Hafnia Pantoea Kluyvera Cedeca Ewingella Moellerella Leclercia Canella Yokenella IAFP, Berlin, 2009
Enterobacteriaceae: Bioindicator
(ISO TC34/SC9, E. de Boer)
Enterobacteriaceae Detection and enumeration (ISO 21528:2004) Revise part 1 of ISO 21528 by omitting the EE broth enrichment step and to modify the confirmation step (fermentation of glucose has been changed in the
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Virus
(CEN TC275/WG6/TAG4, R. Rangdale (CRL))
Draft of a full standard: Horizontal method for detection of hepatitis A virus and norovirus in food using real-time RT-PCR- Part 1: Method for quantitative determination Norovirus genogroups I and II New target matrices: According to regulation EC 178/2002, bottled water and water used in the food industry is a food.
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Parasites
(ISO TC34/SC9/WG6 and CEN TC275/WG6/TAG7, P. Rossi (CRL))
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METHODS
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Revision of EN ISO series : Preparation of test samples, initial suspension and decimal dilutions for microbiological examination Agreement with the development standards about sampling techniques.
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of
General requirements relating to Molecular methods for detection and quantification of microorganisms
(CEN TC275/WG6/TAG3, K. Berghof)
Standards: General rules for PCR and validation of thermocycler New project standards for Real Time - PCR
polymerase chain reaction (PCR) for the detection of food-borne pathogens -- General requirements and definitions
EN ISO/DIS 22118 Microbiology of food and animal feeding stuffs -- Polymerase chain reaction (PCR) for the detection of food-borne pathogens -- Performance characteristics of molecular detection methods
The performance characteristics of molecular methods shall be established according to the procedures of EN ISO 16140 if applicable
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Pathogenic Vibrio parahemolyticus Detection and enumeration on account of poor performance of cultural methods ISO 21871 and 21872 toxR gene sequence R72H and hemolysin genes TDH and TRH
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Necessity of General rules and guidelines Necessity for detection of staphylococcal enterotoxins
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Quality management
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Culture media
(ISO/TC34/SC9/WG5 and CEN TC275/WG6/TAG1, M. Greenwood) Revised EN ISO 11133-1 had been published in February 2009 again as a TS: Microbiology of food and animal feeding stuffs - Guidelines on preparation and production of culture media - Part 1 : general guidelines on quality assurance for the preparation of culture media in the laboratory. Joint standard water and food 2 parts of ISO 11133 will become 1 standard pr EN ISO 11133 (Preparation, production, storage and performance testing of culture media) Joint standard water and food Scope extended to reagents and confirmatory media
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Validation
( ISO/TC34 SC9/WG3 and CEN TC275/WG6/TAG2, P. Int Veld) - PG1: Terminology - PG2: Validation of proprietary methods (quantitative and qualitative methods) - PG3: Intermediate validation - PG4: Method verification - PG5: In-house validation (against and without a reference method) - PG6: Technical requirements for establishment/revision of standardized methods
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Validation
( ISO/TC34 SC9/WG3 and CEN TC275/WG6/TAG2, P. Int Veld)
Work First draft: NWIP EN ISO 16140-1 Terminology of Method Validation NWIP EN ISO 16140-2 Protocol for the validation of alternative method Part 2 : Protocol for the validation of propietary methods against a reference method Link with ISO TC34/SC9/WG2 Statistics Revision of EN ISO 16140:2003 Microbiology of food and animal feeding stuffs - Protocol for the validation of alternative methods
Circulation of the draft of project of Amendment A1 of EN ISO 16410:2003 which correct errors in clause 6.3. Extension of its scope to PCR and Immunological methods
EN ISO standards perhaps recognised by AOAC (Paul WEHLING, AOAC, Chairman Statistical Committee, new member of WG2. A full international validation process of microbiological alternative method EN ISO 16140 and the life of reference method? IAFP, Berlin, 2009
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Reference methods
Validation, Equivalence
Alternative methods
Vibrio cholerae
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Proficiency testing
(ISO TC34/SC9/WG4, S. Passmore)
Final vote: CEN ISO/TS 22117 Proficiency testing Request for ILAC and EA recommending them to recognise this TS (Technical Specification) as technical requirements for proficiency testing organisation in the field of food microbiology (DIS ISO 17043).
IAFP, Berlin, 2009
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Measurement uncertainty
(ISO TC34/SC9/WG2, B. Lombard)
New Work:
uncertainty of sampling in food microbiology, maximum allowable values for measurement uncertainty
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Cover the food chain Integrate new technology Improve the publication delay of standards More flexible Avoid to put method inside regulation Financial support for ring trials ! Description of method or standard based only on performance criteria?
IAFP, Berlin, 2009
Acknowlegments
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