Effective label-friendly food protection in brined white cheese

TP 2022-1e

Poster presentation at 5th IDF Symposium on Cheese Ripening held in Bern, Switzerland, March 2008, by D. Kuckelsberg*, E. Vesterlund*, M. Hassing and A. Gravesen*, Sales and Application Food Ingredients, Danisco A/S*.

INTRODUCTION

White cheese samples in various commercial formats: vacuum packed, in brine, in oil

The shelf life of packed white brined cheese is predominantly limited by the development of sensory defects such as a yeasty taste or inflated packages due to gas formation. Chemical preservation is not generally approved and may even be forbidden, as is the case with FETA (DOP). The objective of this study was to identify the typical shelf-life challenging species and demonstrate the efficacy of Daniscos HOLDBAC YM-B and HOLDBAC YM-C Protective Cultures in brined white cheese applications.
DEVELOPMENT OF PROTECTIVE

D. hansenii Candida famata Greek feta and brine

CULTURES

S. cerevisiae Candida robusta Bulgarian white cheese

K. lactis Candida sphaerica Turkish and Greek white cheese

Species of bacteria that are generally recognised as safe (GRAS) and part of the natural flora of the cheese are particularly interesting candidates for the development of protective cultures. Danisco applies Lactobacillus rhamnosus, Lactobacillus paracasei and Propionibacterium freudenreichii in a range of formulations.
IDENTIFICATION OF TYPICAL SPOILAGE FLORA

Y. lipolytica Candida lipolytica Greek feta

Unidentified species Turkish white cheese and brine

Samples of commercially available cheese from Greece, Bulgaria and Turkey were analysed. The yeast flora were isolated and the species identified using REP-PCR (figure 1). The most frequently appearing species were Debaromyces hansenii, Sacharomyces cerevisiae, Klyveromyces lactis and Yarrowia lipolytica.
PROTECTIVE CULTURES IN APPLICATION TESTS

Figure 1: REP-PCR sequencing and dendrogram of yeasts isolated from Greek, Bulgarian, Turkish and Dutch white cheese samples purchased in supermarkets plus reference strains from strain collections.

The efficacy of the protective cultures was tested in an in vitro model system on certain indicator species. HOLDBAC YM-B demonstrated an effective inhibi-

tion spectrum against yeasts (Rhodutorula rubra, Candida quilliermondi / Pichia quilliermondii) and moulds (Aspergillus niger, Penicillium spp.). HOLDBAC YM-C was found to inhibit moulds (Penicillium sp.) and provided an inhibitory profile against a range of yeast species (Candida krusei / Issatchenkia orientalis, Candida magnoliae, Candida parapsilosis, Candida pulcherrima / Metschnikowia pulcherrima, Candida reukauffii, Candida silvicola, Sporobolomyces sp., Zygosaccharomyces sp.). However, the most relevant results are obtained from in vivo trials using realistic plant conditions. Two different approaches were taken. White cheese samples were produced under industrialscale conditions and either the protective capacity against a naturally occurring contamination was investigated, or the surface of the samples was contaminated intentionally with a defined contamination load. In both cases, the samples and reference were stored and analysed.
WHITE CHEESE PROTECTION AGAINST MOULD

Sample A

mimic the standard fridge temperatures of North European households. After being subject to temperature abuse at 25C, even cheeses with the low contamination rate of 1E+01 spores per spot turned mouldy within 5 days.
WHITE CHEESE PROTECTION AGAINST YEAST

Contaminated at 3 spots with 1E+01 cfu/ spot and incubated at 10C for 7 days. Sample B

Contaminated at 3 spots with 1E+04 cfu/ spot and incubated at 10C for 5 days. Sample C

Test results White cheese from industrial-scale products were chopped and contaminated with defined amounts of Penicillium sp. spores (isolated from white cheese of Lebanese origin). To the left in each set of cups is reference cheese from unprotected cheese. To the right in each set is cheese protected with HOLDBAC YM-B at a dosage of 5 DCU per 100 litres of vat milk. The circles mark mould colonies.The spores were applied at a level of 10l suspension/spot.

Contaminated at 3 spots with 1E+01 cfu/ spot and incubated at 25C for 5 days. At low recontamination rates (here 10 spores per spot), the food matrix gained sufficient protection from the pH of 4.5-4.7 and typical salt levels of 2.22.3% NaCl. At elevated contamination rates, the unprotected cheese stored at 10C was spoilt by mould colonies on day 5. A temperature of 10C was chosen to

Test results White brined cheese was produced at industrial scale, and the protective cultures added to the vat milk together with the acidifying starter culture. An increased dosage of the protective culture from 5 to 10 DCU per 100 litres of vat milk delayed cheese deterioration by 8 to 9 weeks (figure 2). Initial counts of Klyveromyces lactis were below the detection level of 1E+02 cfu/g cheese but grew to >1E+05 cfu/g within less than 3 months in the unprotected cheese (figure 3). Both protective cultures, HOLDBAC YM-B and HOLDBAC YM-C inhibited growth at a dosage of 5 DCU per 100 litres, HOLDBAC YM-C being the most effective. When HOLDBAC YM-C was added, yeast counts increased to 3E+02 cfu/g cheese for 4 months, then dropping to the detection level until 8 months and, after that, to below the detection level until the end of the 12-month observation period. Greek feta cheese is traditionally expected to have a 12-month shelf life.
PAINS, FEATURES, GAINS

The primary pain in white cheese technology is the potential deterioration of sensory aspects due to the growth of yeasts and/or moulds. Contaminants can grow to levels of >1E+5 cfu/g cheese, even under cold storage conditions.

REFERENCES
1.0E+07 1.0E+06 1.0E+05

1.0E+04 1.0E+03 1.0E+02 1.0E+01 0 2 4 6 8 10 detection limit

Weeks
HOLDBAC YM-B (5 DCU/100l)

HOLDBAC YM-B (10 DCU/100l)

No HOLDBAC

Figure 2: Cheese surface contaminated with 1E3 cfu D. hansenii/g cheese after re-packing in brine. The protective culture had been added to the vat milk at 5 DCU per 100 litres and 10 DCU per 100 litres vat milk respectively for the trial products.The cheese was stored in brine at 8C in closed plastic cups and at an ambient temperature.

1.0E+07 1.0E+06 1.0E+05

Cfu/g

1.0E+04 1.0E+03 1.0E+02 1.0E+01 0 2 4 6

Months
HOLDBAC YM-B (5 DCU/100l)
HOLDBAC YM-C (5 DCU/100l)
No HOLDBAC

T.H. Suamalainen, A.M. Myr-Mkinen: Propionic acid bacteria as protective culture in fermented milks and bread, Le Lait (1999) 79, 165-174. Myr-Mkinen et al.: Lactobacillus casei ssp. rhamnosus, bacterial preparations comprising said strain, and use of said strain and preparations for the controlling of yeasts and moulds, 1995, Patent number 5, 378, 458. Susanne Miescher Schwenninger and Leo Meile: A Mixed Culture of Propionibacterium jensenii and Lactobacillus paracasei subsp. paracasei inhibits Food Spoilage Yeasts, System. Appl. Microbiol. (2004) 27, 229-237. S. Miescher Schwenninger, L. Meile: European Patent Application. Publication no, EP 1308506. Morten Hassing: Application of protective cultures in feta production, Master Thesis at the University of Copenhagen, 2007.

Cfu/g

detection limit 8 10 12 14

Figure 3: Cheese from industrial-scale trial spoilt by Klyveromyces lactis, a wild contamination based on natural background flora.The protective culture was added to the vat milk at 5 DCU per 100 litres.The cheeses were stored at 4C in sealed deep pull packages.

Two effective HOLDBAC YM protective cultures each display a natural inhibition capacity against yeast and mould species. The antimycotic effect is a result of competitive exclusion and a combination of non-specific metabolic by-products. Propionic acid was not detectable in the cheese. The protective cultures inhibit the growth of contaminants but do not lyse or inactivate contaminants.

HOLDBAC YM cultures can provide a label-friendly solution to shelf life challenges. The inhibitory capacity depends on the species combined in the protective culture as well as a dose response pattern. Protective cultures give a higher degree of certainty that a 12-month shelf life will be achieved. No adverse sensory effects were observed during the study.

Danisco A/S Edwin Rahrs Vej 38 DK-8220 Brabrand, Denmark Telephone: +45 89 43 50 00 Telefax: +45 86 25 10 77 info.ingredients@danisco.com www.danisco.com

The information contained in this publication is based on our own research and development work and is to the best of our knowledge reliable. Users should, however, conduct their own tests to determine the suitability of our products for their own specific purposes and the legal status for their intended use in the product. Statements contained herein should not be considered as a warranty of any kind, expressed or implied, and no liability is accepted for the infringement of any patents.

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