Introducing Innovative Diagnostics and HACCP in the Ornamental Fish Industry: sense or non-sense?

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1 Introducing Innovative Diagnostics and HACCP in the Ornamental Fish Industry: sense or non-sense? Prof. Kris A. Willems, PhD and Bart Lievens, PhD, Ir. Scientia Terrae Research Institute (STRI), Belgium De Nayer Institute, K.U.Leuven Association, Belgium Interzoo, OFI Fish Health Seminar May 23 th, 2008

2 Quality Driving Force in the Market Diversity of systems ISO 9000, EFQM, TQM GAP, GMP, GLP, GEP Process control & improvement Systematic approach Appropriate resources & best available technologies Traceability Total approach Continuous improvement Throughout organisation and/or food supply chain All aspects of supplier-customer relationship Quality Satisfy the customer s needs at the best economical conditions Product oriented Process oriented Total approach

3 Quality Systems Process Control - Core of all Quality Systems resources people instructions registrations assignment requirements (production) process product corrective actions measurements control limits specifications

4 Food Safety Focus on Human Health and/or Spoilage Chemicals Micro-organisms Fysical agents Effect Intoxication Infection Intoxication Wounds Not acceptable Discolouring of ham Fungi on bread Hair in burger Unsafe pesticides L. monocytogenes in salad Glass in burger

5 Ornamental Fish Industry (OFI) Focus on Disease Prevention Biosecurity Prevention of the entry of new or unwanted organisms, especially infectious agents Biosecurity includes all measures in place to protect ornamental aquatic organisms (fish, invetabrate or plant) from contracting, carrying or spreading disease Prevention Detection Management, i.e. control and reduction

6 Food Safety Definition of HACCP Hazard Analysis Critical Control Points: A systematic approach to the identification and assessment of the microbiological hazards and risks associated with the manufacture, distribution and use of a particular foodstuff and the definition of means for their control International Commission on Microbiological Specifications on Foods - ICMSF

7 Food Safety Primary Characteristic of Food Quality Prevent disease Legal requirement post-farm Core HACCP concept Diversity of systems HACCP BRC, IFS ISO General Food Law (EU) Process Control & Improvement Quality Food Safety Hazard analysis Principles 1 & 2 Control loop Principles 3, 4 &5 Documentation & Verification Principles 6 & 7

8 HACCP Essential Elements Hazard Analysis & Process Control Identify hazards and assess severity Determine Critical Control Points (CCP s) Specify control criteria, i.e. limits and tolerances Monitor Critical Control Points Identify and take corrective action(s) Verify the system is working Keep records Process Control & Improvement Quality Food Safety Hazard analysis Principles 1 & 2 Control loop Principles 3, 4 &5 Documentation & Verification Principles 6 & 7

9 HACCP Characteristics Systematic approach to identify those factors that directly affect food safety / biosecurity Preventive instead of retrospective approach Replaces traditional inspection and testing regimes More cost-effective control system Concentration resources, e.g. technical and financial, into critical areas No cure all Absolute safety is absolutely unattainable

10 General Food Law EU Hygiene Directive General rules for the production of all foodstuffs Specific rules for meat, fish, dairy, eggs, snails and frogs, animal fats, gelatine and callogen Implementation a HACCP is a legal requirement, except for the primary sector Primary sector => Good Agricultural Practices (GAP) Product liability confirmed, now including slaughterhouses and animal wellfare

11 Terminology From Hazard to Risk (1) Hazard Food safety - Biological, chemical or fysical agent that potentially can affect the safety of a foodstuff Biosecurity Biological agent that potentially can affect fish health, human health and/or ecosystems Hazard Analysis Process of collecting and evaluating information on hazards and conditions leading to their presence to decide which are significant for food safety and therefore should be addressed in the HACCP plan

12 Terminology From Hazard to Risk (2) Risk Probability x Consequences = Risk Risk Analysis A process consisting of three components: Risk assessment Risk management Risk communication Advocated by many governments and intergovernmental organisations Triggered by World Trade Organisation (WTO)

13 Terminology From Hazard to Risk (3) Risk Assessment - Hazard Identification - Hazard characterisation - Exposure assessment - Risk characterisation Risk Management - Risk evaluation - Option assessment - Option implementation - Monitoring & review Risk Communication Interactive exchange of information and opinions concerning risks

14 Terminology From Hazard to Risk (3) Risk Assessment A scientifically based process consisting of the following steps: 1. hazard identification; 2. hazard characterisation; 3. exposure assessment; 4. risk characterisation Risk Management The process of weighing policy alternatives, considering risk assessment and other factors relevant for the health of consumers and for the promotion of fair trade practices Risk Communication The interactive exchange of information and opinions throughout the risk analysis process concerning risk among risk assessors, risk managers, consumers, industry, the academic community and other interested parties.

15 Risk Analysis The Broader Context Country level Operational level Policy Standards Specific Hazard Management HACCP Risk Analysis - high level - policy based guidance - specific standards, criteria Food Safety Management - local - specific management at supply chain level Generic Hazard Management GHPs / GMPs / GAPs

16 Good Practices & HACCP Foundation of Food Safety / Biosecurity Good Practices Foundation of all (food) safety systems General guidance on sanitary practices and level of care of (food manufacturing) facilities HACCP Applies to a specific product, manufactured on a specific location and/or production line Specificity and stringency of hazard control essential to its success Stringency currently not really defined in quantitative terms

17 Good Practices & HACCP Require Technical and Scientific Data Epidemiological data on (microbial) pathogens Incidence of foodborne illness Surveillance programmes Raw material, intermediate and final product data ph, a w, presence of preservatives, packaging materials and conditions, product structure, processing conditions, shelf life, etc Processing data Number sequence of processing data, range of process parameters, efficacy of cleaning and disinfecion Microbiological data Likely presence of microbiological hazards in raw materials, growth and death rates under processing conditions, etc..

18 Good Practices & HACCP Critical Success Factors Technical and scientific skills of (food) business operator Regulatory guidance regarding key operations, (i.e. desinfection, pasteurisation, etc. Regulatory guidance on safety benchmarks, microbiological criteria, etc.

19 Risk Analysis / Risk Assessment Microbial Risk Assessment (MRA) Purpose Systematic assessemnt (model/estimate) of the level of risk prevailing in a country as associated to pathogenic micro-organisms (in foods) Inventorisation of typical risk contributing and risk mitigating factors Elaboration of possible risk mitigation strategies Scope All foods consumed in a specific country, whether produced or imported Outcome Estimated number of cases of (a certain) illness per year per (e.g.) persons in a given population caused by a certain micro-organism or group of micro-organisms in a particular food product or food type Chance of illness due to consumption of a specific food product (per serving event) to which a particular hazard can be associated

20 Risk Analysis / Risk Assessment Microbial Risk Assessment (MRA) Role of (Food) Business Operators Do not need to conduct risk assessments Need to ensure proper implementation of (food) safety management systems, incl. HACCP MRA studies can strenghten (food) safety sysyems Relevance to (Food) Business Operators Improvement of GHP & HACCP systems by Learning about differences in levels of hazard control between (typical) operations Appreciating different intervention strategies or management options Seeing examples of how to meet government requirementq (incl. (food) safety objectives when mandated)

21 Quality Systems Process Control - Core of all Quality Systems resources people instructions registrations assignment requirements (production) process product corrective actions measurements control limits specifications

22 Biosecurity Fish Disease Management Main fish pathogens: bacteria, fungi, viruses, and protozoa Disease management Accurate diagnosis Choice of appropriate process control and management strategies Preventive disease management

23 Biosecurity Main Limitations of Fish Disease Management Lack of rapid and accurate detection and identification of fish pathogens Standard versus alternative methods

24 Biosecurity / Microbiological Methods Traditional Methods Wide diversity of assays Time consuming Labour intensive Specialist laboratories

25 Biosecurity Main Limitations of Fish Disease Management Lack of rapid and accurate detection and identification of fish pathogens Standard versus alternative methods Multiplexing and quantification

26 Biosecurity Main Limitations of Fish Disease Management Lack of rapid and accurate detection and identification of fish pathogens Standard versus alternative methods Multiplexing and quantification Most fishes can be infected by a multitude of pathogens Similar symptoms may be caused by different pathogens Disease symptoms often result from infection by multiple pathogens Revealing relevant (pathogen/host) associations

27 Biosecurity Main Limitations of Fish Disease Management Lack of rapid and accurate detection and identification of fish pathogens Standard versus alternative methods Multiplexing and quantification Infectious threshold Action threshold

28 Biosecurity / Microbiological Methods Alternative (Molecular) Methods Serological and nucleic acid- (DNA/RNA) based techniques Generally faster, more specific, more sensitive and more accurate than conventional methods Equally suitable for the detection of culturable as well as non-culturable micro-organisms No experienced taxonomists required However: detection limited to a single or a few pathogens

29 Biosecurity / Microbiological Methods Molecular Methods: DNA arrays Hybridization of labeled target amplicons to specific DNA fragments bound to a solid support (macroarrays vs microarrays) Combines PCR amplification with the unlimited screening capability of DNA arrays: high degrees of sensitivity, specificity, and throughput capacity Reduces costs and increases speed of diagnosis Successful application to detect plant pathogens: DNA Multiscan

30 Biosecurity / Microbiological Methods DNA arrays: different approaches of detection Universal Primer based multiplex detection Discrimination of phylogenetically related organisms (e.g. bacteria and fungi) Random Primed based multiplex detection Discrimination of phylogenetically less related organisms (e.g. viruses)

31 Biosecurity / Microbiological Methods DNA arrays: results read as a checklist Microorganisms present in the sample are literally highlighted Compared to reference oligonucleotides the intensity of the hybridization signal is an indicator for the amount of biomass present (semi-quantification)

32 Biosecurity / Microbiological Methods DNA arrays: multiple detection in 36 hours 36 hours

33 Biosecurity / Microbiological Methods DNA arrays: an example Detection of multiple pathogens Still expertise required to interpret results Aeromonas sp. A. salmonicida Flavobacterium sp. F. branchiophilum Mycobacterium sp. M. marinum Universal All bacteria All oomycetes S. cerevisiae Calibration control

34 Biosecurity / Microbiological Methods DNA array OFI - Target Organisms Bacteria: Aeromonas hydrophila, A. salmonicida, Edwardsiella ictaluri, Flavobacterium branchiophilum, F. columnare, F. psychrophilum, Mycobacterium chelonae, M. fortuitum, M. marinum, Piscirickettsia salmonis, Pseudomonas anguilliseptica, Renibacterium salmoninarum, Streptococcus iniae, Tenacibaculum maritimum, Vagococcus salmoninarum, Vibrio alginoyticus, V. anguillarum, V. parahaemolyticus, V. vulnificus, Yersinia ruckeri DNA viruses: Koi Herpes Virus (KHV), Carp Pox Virus (CPV), Channel Catfish Virus (CCV) RNA viruses: Spring Viraemia of Carp Virus (SVCV), Viral Haemorrhagic Septicemia Virus (VHSV), Infectious Hematopoietic Necrosis Virus (IHNV), Infectious Salmon Anemia Virus (ISAV)

35 Biosecurity / Microbiological Methods DNA array OFI - Applications Analysis of different kind of samples including tissue, water, fish nutrition, and water plants Routine diagnosis of fish diseases Ecological and epidemiological studies Sectoral surveillance in support of HACCP & GMP Allows taking well-informed decisions in terms of biosecurity and a safe ornamental fish trade

36 Biosecurity / Microbiological Methods Validation of Alternative Methods Essential steps required to evaluate new technologies rarely taken Microval => Harmonised EU validation procedure Validation: Specificity Sensitivity Reproducibility Accuracy of results Ring tests Challenge: Singleplex vs multiplex assays?

37 Acknowledgements Scientific support Scientific advisors Dr. B. Austin, Heriott-Watt University, UK Dr. I. Pavlik, Veterinary & Pharmaceutical University Brno, Czech Republic Dr. P. Silley, University of Bradford, UK Dr. F. Lieffrig, Centre d Economie Rurale, Belgium Dr. B. Thomma, Wageningen University, the Netherlands Dr. P. Sorgeloos, Ghent University, Belgium Other Dr. M. Snow and Dr. A. McBeath, Fisheries Research Services, Marine Laboratory, UK Dr. T. Nishizawa, Hokkaido University, Japan Dr. N. Gagne, Fisheries and Oceans Canada, USA Dr. K. Garver, Fisheries and Oceans Canada, Pacific Biological Station, USA Dr. M. Sakai and Dr. T. Kono, University of Miyazaki, Japan Dr. T. Iida, Fisheries Research Agency, Japan Dr. G. Kurath and Dr. M. Purcell, Western Fisheries Research Center, USA Dr. M. Kotler, Department of Pathology, Israël Dr. P.Y. Lee, Graduate Institute of Medical Biotechnology, Taiwan Dr C. Amaro, Department of Microbiology and Ecology, Spain

38 Acknowledgements Financial support Vlaams Instituut voor de bevordering van het Wetenschappelijk- Technologisch Onderzoek Vlaanderen, Belgium Industrial sponsors Wholesalers: Bassleer Biofish nv., Aquaria Antwerp bvba., De Jong Marinelife bv. Fish nutrition: INVE Technologies nv., Fish breeder and fish hatchery: Joosen-Luyckx Aqua Bio nv. Veterinary Surgeons: Dr. S. Verbinnen, Dr. T. Barbé, Dr. M. Lammens, Dr. L. Lambrechts, Dr. S. Teerlinck Ponds and Aquarium Specialist Shop: Aqua Hobby nv. Diagnostic company: BioART nv. Testing Laboratory: MicroBioMetrix bvba. International Trade Association: Ornamental Fish International Disinfection Water Supply Systems: Uniprox Europe BV

39 Introducing Innovative Diagnostics and HACCP in the Ornamental Fish Industry: sense or non-sense? It makes a lot of sense! Thank you Questions?

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