Danisco A/S FOOD PROTECTION SYMPOSIUM SAO PAULO, BRAZIL JUNE 2005 Natural Food Safety in the DAIRY WORLD Steffen Thun Application Technologist Dairy Applications, Germany HOLDBAC™ NATAMAX ® MicroGARD™ NISAPLIN ®

Danisco A/S Natural Food Safety in the DAIRY WORLD Food preservation Organoleptic quality maintenance of food product throughout its shelf life Food safety Food safety Reduction of the risk of pathogens in food product throughout its shelf life Food protection

Danisco A/S 400 Dairy Spreads, Cheese Fillings 300 Flavoured Milks Natural/processed Cheese 100 Cottage Cheese Yoghurt Sour Cream MicroGARD™ 100, 300, 400 Fermentation Metabolites CMP 510 Dairy Spreads Flavoured Milks 200 Cottage Cheese Yoghurt Sour Cream MicroGARD™ 200, 510 Fermentation Metabolites CSP MicroGARD™ Fermentates Gram-neg.& Gram-pos. spoilage Yeasts & Moulds NATAMAX® additionally Yoghurt, Sour Cream Cottage Cheese etc. Salt & SF Shredded cheese Cheese Coating NATAMAX® Standard, Salt, SF Natamycin NISAPLIN® Cheese Neutral Dairy Products Direct acidified MP NISAPLIN® Nisin NATAMAX® Natural Antimicrobial Yeasts & Moulds YM-C Yoghurt, Fresh Cheese Sour Cream Cottage cheese etc. YM-B Yoghurt, Fresh Cheese Sour cream Cottage Cheese etc. YM-B, YM-C Lactic bacteria & Propionibacteria LC Salt brine Semi-hard/White Cheeses Listeria Soft, blue, semi-hard cheeses HOLDBAC™ Listeria, LC Lactic bacteria HOLDBAC™ Protective Cultures Yeasts & Moulds Listeria, Heterofermentative Lactobacilli Food Protection Solutions in Dairy Applications offered by DANISCO Natural Food Safety in the DAIRY WORLD NISAPLIN® Natural Antimicrobial Gram-positive Bacteria HOLDBAC™ NISAPLIN® Milk Free All applications

Danisco A/S HOLDBAC™ YM Protective Cultures against Yeasts and Moulds Natural Food Safety in the DAIRY WORLD

Danisco A/S HOLDBAC™ – food safety goes live QMS GMP HACCP- Concept Bio Preservation

Danisco A/S Micro-organisms „Protective Cultures“ Cultures“ Food final products pre-stages or raw material Inhibition of foodInhibition of food pathogenic micro- organisms growth control prevention of toxin- formation reduction of pathogens and/or Shelf lifeShelf life extension extension based on targeted inhibition of specific spoilage micro-organisms HOLDBAC™ – food safety goes live

Danisco A/S HOLDBAC ™ - What makes them anti-microbial? Anti-microbial metabolites Organic acids lactic acid acetic acid propionic acid… Hydrogen peroxide Carbon dioxide Lacto peroxide Fatty acids Diacetyl Acetaldehyde Reuterin Other compounds of low molecular mass Bacteriocins

Danisco A/S HOLDBAC ™ - more than anti-microbial metabolites! anti-microbial metabolites competitive exclusion effects further, so far unknown effects (e.g. quorum sensing) Inhibition of undesired micro-organisms by competing for nutrients, oxygen, etc. = + + Inhibition of food pathogenic and food spoilage micro-organisms

Danisco A/S HOLDBAC ™ cultures - a natural way Safer food = Bio preservation by protective cultures: Label friendly – It´s a „culture“ and not a food preservative = Positive image of „biopreservation“- „natural“ =

Danisco A/S Protective Cultures - categories PROTECTIVECULTURE STARTERCULTURE Starter Cultures Adjunct-Cultures Multi-functional Cultures Non-fermenting Protective Cultures fermentation processes: Changes of taste, flavour and texture no influence on sensory characteristics

Danisco A/S Protective Cultures - categories PROTECTIVECULTURE STARTERCULTURE Starter Cultures Adjunct-Cultures Multi-functional Cultures Non-fermenting Protective Cultures fermentation processes: Changes of taste, flavour and texture no influence on sensory characteristics

Danisco A/S Protective Cultures - categories PROTECTIVECULTURE STARTERCULTURE Starter Cultures Adjunct-Cultures Multi-functional Cultures Non-fermenting Protective Cultures fermentation processes: Changes of taste, flavour and texture no influence on sensory characteristics

Danisco A/S Protective Cultures - categories PROTECTIVECULTURE STARTERCULTURE Starter Cultures Adjunct-Cultures Multi-functional Cultures Non-fermenting Protective Cultures fermentation processes: Changes of taste, flavour and texture no influence on sensory characteristics

Danisco A/S Protective cultures - are safe generally recognised as safe (GRAS) Protective cultures are: there is no indication of a health risk of this bacterial group the use of lactic acid bacteria in biological preservation might even contribute to the health benefits of a product * ) Lactic acid bacteria Lactobacillus sp. Lactococcus sp. Propionibacteria * ) * ) Holzapfel et al., 1995, Int. J. Food Microbiol. 78:

Danisco A/S HOLDBAC™ HOLDBAC™ Cultures - 4 steps from our lab to our customers STEP 1: Isolation Selection Screening STEP 2: Process- Development Up-Scaling Down-Stream- Processing STEP 3: Application Application tests in food model systems STEP 4: Challenge tests Tailor made „friendly label“ HOLDBAC™ applications in food & feed

Danisco A/S Food Model Systems Preparation Fermentation / Ripening Storage Pack- aging Continuous Monitoring: Behaviour of indicator strains (growth – inhibition) Influence of the Food Protectants on technological and organoleptic properties in real food matrixes Defined contamination with L3* food pathogens (e.g. Cl. botulinum, Campylobacter sp., E. coli H157,…) Food spoilage microorganisms Microbiological Flora Analyses Sensory Evaluation Physico-chemical Analyses Technological Properties End of shelf life To reproduce the food products from production to the end of shelf life: