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Biopreservation of Anaerobically Packaged Sliced Cooked Meat Products by Non-Bacteriocinogenic Micro-Organisms

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Biopreservation of Anaerobically Packaged Sliced Cooked Meat Products by Non-Bacteriocinogenic Micro-Organisms Promotors: Prof. dr. ir. Johan Debevere Prof. dr. ir. Frank Devlieghere Laboratory of Food Microbiology and Food Preservation Department of Food Safety and Food Quality Faculty of Bioscience Engineering Ghent University Dean: Prof. dr. ir. Herman Van Langenhove Rector: Prof. dr. Paul Van Cauwenberghe ir. Lieve Vermeiren Biopreservation of anaerobically packaged sliced cooked meat products by non-bacteriocinogenic micro-organisms Thesis submitted in fulfillment of the requirements for the degree of Doctor (Ph.D.) in Applied Biological Sciences Bioconservering van anäeroob verpakte versneden gekookte vleesproducten met niet-bacteriocineproducerende micro-organismen Illustratie: Lactobacillus sakei 10A © Lieve Vermeiren Bacteriophage P200 © Steven Hagens (Swiss Federal Institute of Technology) Vermeiren, L. (2006). Biopreservation of anaerobically packaged sliced cooked meat products by non-bacteriocinogenic micro-organisms. Doctoral dissertation. Universiteit Gent. ISBN: 90-5989-132-5 The author and the promotor give the authorisation to consult and to copy parts of this work for personal use only. Every other use is subject to the copyright laws. Permission to reproduce any material contained in this work should be obtained from the author. Woord vooraf Proefbuizen, labojassen, cellen, microscopie, micro-organismen,… het fascineerde mij al heel vroeg. Het willen weten, onderzoeken, analyseren en trachten oplossen, dat heeft er altijd al wel ingezeten. Na mijn thesis aan het Labo Levensmiddelenmicrobiologie en –conservering was ik zo gebeten door de labo-microbe, letterlijk en figuurlijk, dat ik dan ook met groot enthousiasme gevraagd heb aan Prof. dr. ir. J. Debevere om verder te blijven werken aan zijn labo. Daar is het uiteindelijk allemaal mee begonnen en ik ben Prof. dr. ir. J. Debevere zeer dankbaar voor die eerste kans maar uiteraard ook voor al de daaropvolgende kansen, die ik telkens kreeg, als assistente en ook nu weer in mijn toekomstige job. Professor, je interesse, je kritische opmerkingen en je bewonderenswaardige ervaring en kennis hebben sterk bijgedragen tot het tot een goed einde brengen van dit werk. Prof. dr. ir. Frank Devlieghere of mag ik Frank zeggen, jij hebt mijn achtjarige carrière het meest van dichtbij gevolgd… en ik ben je extra dank verschuldigd. Al van in mijn thesistijd stond je deur altijd open. Ondanks je drukke agenda, wist je tijd vrij te maken voor het helpen oplossen van alledaagse onderzoeksprobleempjes, voor het grotere brainstormwerk op de kritieke momenten van dit werk en voor het kritisch beoordelen van het schrijfwerk. Je energie en enthousiasme werkten inspirerend. Prof. Debevere en Prof. Devlieghere, bedankt voor het promotorschap op jullie te nemen maar vooral bedankt voor jullie vertrouwen. Mijn werk was nooit alleen mijn werk. Mijn werk was en is ook mijn collega’s… en die groep collega’s werd door de jaren heen steeds groter. Jullie allemaal opnoemen is dan ook onmogelijk (ook al drongen sommigen hierop sterk aan), maar elk van jullie heeft wel hier of daar zijn steentje bijgedragen tot dit werk: wetenschappelijke steun, technische steun, administratieve steun, IT-steun, morele steun … BEDANKT daarvoor. Ook wil ik een zeer groot woord van dank richten aan al de thesisstudenten die meegeholpen hebben aan dit werk: Ali, Veerle, Annelies, Isabelle, Ursula, Barbara. Jullie hebben uiteraard elk een belangrijk deel van het praktische werk voor jullie rekening genomen, maar veel belangrijker voor mij was dat jullie aanwezigheid mij het gevoel gaf er niet alleen voor te staan, het samen aan iets kunnen werken, het samen naar oplossingen zoeken, samen bloed, zweet en tranen vergieten,… was uiterst motiverend. Dit doctoraat afleggen was een zware reis… en naast de wetenschappelijke kennis die ik opbouwde, leerde ik eigenlijk veel meer andere dingen… zoals geduld, volharding en relativeringsvermogen, maar niet zonder hulp van mijn gezin en mijn familie. Aan mijn ouders en schoonouders kom ik woorden van dank tekort. Het is ongelooflijk hoe onvoorwaardelijk en nooit aflatend jullie steeds hulp aanbieden en geleidelijk aan, naarmate dit werk vorderde en de nood eraan toenam, leerde ik ook beter die hulp te aanvaarden. Tom, jij hebt alle fasen van dit doctoraat mee beleefd… de perioden van mist, optrekkende mist en helder weer. Jij bent de enige die alle hoekjes van mijn ziel kent. Je laat me zijn zoals ik ben en dat is heel veel waard…. Bovendien schonk je mij twee schatten van zonen en dat ‘mama’ worden heeft ook op zijn manier bijgedragen tot dit werk… het plaatste dit doctoraat in de juiste context… en zo leerde ook ik stilaan relativeren. Jorne en Brent, ook aan jullie een woordje. Nu zijn jullie nog veel te klein om dit te begrijpen maar ooit komt een tijd dat jullie dit wel zullen lezen. Het was niet altijd gemakkelijk om dit doctoraatswerk te combineren met mijn zorg voor jullie maar ik wil dat jullie weten dat jullie steeds op de eerste plaats stonden en dat ook altijd zo zal zijn. Lieve 26 september 2006 Table of contents Table of Contents Table of Contents .................................................................................... i Introduction and objectives ................................................................ vii Summary................................................................................................ xi Samenvatting....................................................................................... xvi Chapter 1: Antagonistic micro-organisms for biopreservation of food products ......................................................................................... 1 1. LACTIC ACID BACTERIA............................................................................................... 1 1.1. INTRODUCTION 1 1.2. TAXONOMY 1 1.3. METABOLISM AND ENERGY GENERATION 2 1.4. LACTIC ACID BACTERIA OF IMPORTANCE IN FOOD PRODUCTS 4 1.4.1. The genus Lactococcus............................................................................................. 4 1.4.2. The genus Leuconostoc............................................................................................. 4 1.4.3. The genus Pediococcus............................................................................................. 5 1.4.4. The genus Lactobacillus........................................................................................... 5 1.5. ANTIMICROBIAL ACTIVITY OF LACTIC ACID BACTERIA 8 1.5.1. Fermentation end-products ....................................................................................... 9 1.5.2. Bacteriocins ............................................................................................................ 13 1.5.3. Other antagonistic systems ..................................................................................... 17 2. THE MICROBIAL ECOLOGY OF ANAEROBICALLY PACKAGED SLICED COOKED CURED MEAT PRODUCTS............................................................................. 18 2.1. PRODUCTION PROCESS 18 2.2. CONTAMINATION 18 2.3. MICROBIAL SPOILAGE 19 2.3.1. Factors determining the microbial growth in CMP ................................................ 19 2.3.2. Specific spoilage organisms ................................................................................... 19 2.3.3. Spoilage phenomena............................................................................................... 21 2.4. SAFETY 24 2.4.1. Pathogens associated with cooked meat products .................................................. 24 2.4.2. Listeria monocytogenes .......................................................................................... 24 3. BIOPRESERVATION....................................................................................................... 26 3.1. DEFINITION AND PRINCIPLE 26 3.2. BIOPRESERVATION BY MEANS OF MICROBIAL METABOLITES 27 3.2.1. Bacteriocins ............................................................................................................ 27 3.2.2. Organic acids .......................................................................................................... 29 3.2.3. Fermentates............................................................................................................. 30 3.3. BIOPRESERVATION BY MEANS OF MICRO-ORGANISMS 30 3.3.1. Protective LAB ....................................................................................................... 30 3.3.2. Bacteriophages........................................................................................................ 41 Table of Contents i 3.4. EFFECT OF PROTECTIVE LAB ON THE SENSORY QUALITY OF ANAEROBICALLY PACKAGED COOKED MEAT PRODUCTS 46 3.5. MECHANISM OF INHIBITION OF PROTECTIVE LAB IN ANAEROBICALLY PACKAGED COOKED MEAT PRODUCTS 48 3.5.1. Production of inhibitory substances........................................................................ 49 3.5.2. Nutrient competition............................................................................................... 50 3.5.3. Competition for space (attachment/adhesion sites) ................................................ 53 3.5.4. Quorum sensing...................................................................................................... 54 3.6. PRACTICAL IMPLEMENTATION OF PROTECTIVE CULTURES 54 3.6.1. Safety aspects.......................................................................................................... 54 3.6.2. Regulatory aspects.................................................................................................
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