Thesis (MSc)--Stellenbosch University, 2001. / ENGLISH ABSTRACT: Kepi is a refreshing, fermented dairy beverage that has been consumed for centuries and is traditionally made by incubating Kepi grains in milk. The Kepi grain is a complex starter culture consisting of a variety of lactic acid bacteria and yeasts. The successful marketing of the grains requires the effective preservation of the microbes present in the grains as well as an appropriate packaging that will retain the acidification activity of the preserved grains over an extended period of time. The aim of this study was to evaluate different preservation techniques and packaging materials in terms of their respective abilities to retain grain viability and activity over an extended storage period. Four different preservation techniques (freezing at -18°C, refrigeration at 4°C, air-drying and Iyophilisation) and three packaging materials including a low density polyethylene film (LOPE), an oriented polyester film (OPET) and a metallised oriented polyester film (MOPET), were evaluated. Activity tests were used to evaluate the impact of the preservation techniques in terms of the retainment of the acidification activity of the preserved grains, and the storage potential of the preserved and packaged grains. The activity tests included changes in pH, %TA, lactic acid production and lactose and volatile compound content over an 18 h fermentation period. In addition, the microbial viability of the packaged Iyophilised grains after two months of storage, was also investigated. Frozen and refrigerated grains showed the best retainment of the acidification activity over a 10-month storage period. Air-drying and Iyophilisation showed a good retention of the activity up to three months of storage, but the application of these techniques both resulted in a retarded initial acidification activity. After 10 months of storage, the air-dried and Iyophilised grains showed only a low acidification activity. No volatile compounds could be detected during the course of the fermentation period, due to the relative short fermentation period of 18 h. Overall, the best retainment of the fermentation activity was given by the LOPE and the OPET packaging films. However, the storage period had a considerable influence on the retention of the activity of the packaged Iyophilised grains. The viability study of the Iyophilised packaged Kepi grains after two months of storage showed leuconostocs and lactobacilli to be the prevalent microbes in the grains. Low microbial counts were obtained from the lactococciselecting medium for all three of the differently packaged Kepi grains, whereas no growth was observed on the media that selected for the propionibacteria and yeasts. The OPET packaging film provided the best preservation of the microbial composition. It was, therefore, concluded that all four preservation techniques would be suitable for the preservation of Kepi grains and the subsequent storage at room temperature for three months. However, for storage periods of 10 months or longer the use of freezing and refrigeration are recommended as most suitable preservation techniques. All three of the packaging materials proved to be suitable for the packaging and storage of the Iyophilised Kepi grains for periods of up to one month. However, for storage periods of two months or longer, the use of the OPET film for the packaging and retainment of the acidification activity of the Iyophilised grains, can be recommended. / AFRIKAANSE OPSOMMING: Kepi is 'n eeu-oue verfrissende, gefermenteerde suiweldrankie wat tradisioneel vervaardig word deur Kepikorrels in melk te inkubeer. Hierdie Kepikorrels bestaan uit 'n komplekse samestelling van hoofsaaklik melksuurbakteriee en giste. Die effektiewe preservering en verpakking van die korrels is belangrike voorvereistes vir die suksesvolle bemarking daarvan. Dis belangrik dat die preserverinq en die verpakking van die korrels 'n positiewe bydrae sal lewer tot die behoud van die fermentasie-aktiwiteit van die mikrobes in die korrels oar 'n verlengde opbergingsperiode. Die doel van hierdie studie was om die opbergingspotensiaal van verskillend gepreserveerde en -verpakte Kepikorrels te evalueer in terme van die behoud van die lewensvatbaarheid en fermentasie-aktiwiteit van die samestellende mikrobes. Vier verskillende preserveringstegnieke (bevriesing by -18°C, verkoeling by 4°C, lugdroging en vriesdroging) en drie verskillende tipes verpakkingsmateriale, nl. 'n "low density polyethylene film" (LOPE), 'n "oriented polyester film" (OPET) en 'n "metallised oriented polyester film" (MOPET) was qeevalueer. Aktiwiteitstoetsing was gebruik om die impak van die verskillende preserveringstegnieke en die verpakkingsmateriale op die behoud van die fermentasie-aktiwiteit van die Kepikorrels te ondersoek. Die verskillende aktiwitieitstoetse wat gedoen is, het die meting van die verandering in pH, %TA, melksuur- en laktosekonsentrasie oor 'n fermentasieperiode van 18 h ingesluit. Tesame met die aktiwitietstoetsing IS die lewensvatbaarheid van die gevriesdroogde, verpakte Kepikorrels na twee maande van opberging ook ondersoek. Die bevrore en verkoelde Kepikorrels het die beste behoud van aktiwitiet na 'n 10-maande opbergingsperiode getoon. Die gelugdroogde en gevriesdroogde korrels het 'n goeie behoud van aktiwiteit getoon vir 'n opbergingstydperk van tot drie maande, maar beide die lugdroging- en vriesdrogingstegnieke het 'n aanvanklik vertraagde fermentasie-aktiwitieit getoon. Na 'n : opbergingsperiode van 10 maande het beide die gelugdroogde en gevriesdroogde korrels egter 'n lae fermentasie-aktiwiteit getoon. As gevolg van 'n relatiewe kort fermentasieperiode van 18 h kon geen vlugtige komponente in die Kepimonsters gevind word nie. Die LDPE- en OPET-verpakkingsmateriale het die beste behoud van die fermentasie-aktiwiteit van die gevriesdroogde korrels getoon. Die opbergingsperiode het egter 'n aansienlike impak op die aktiwitietsbehoud van die korrels gehad. Die lewensvatbaarheidstudie het aangetoon dat Leuconostoc- en Lactobacillus-spesies die oorheersende mikrobes in die verpakte, gevriesdroogde Kepikorrels na 'n opbergingsperiode van twee maande was. Lae mikrobiese tellings vir al drie van die verpakkingsmateriale was gevind op die Lactococcusselekterende medium, en geen mikrobegroei kon op die giste- en propionibakteriee-selekteringsmedium waargeneem word nie. Die beste behoud van die mikrobiese samestelling in die verpakte, gevriesdroogde Kepikorrels was gevind vir die OPET-verpakkingsmateriaal. Die gevolgtrekking kan gemaak word dat al vier die preserveringstegnieke geskik is vir die preservering van die Kepikorrels en die daaropvolgende opberging van drie maande by kamertemperatuur. Vir opbergingsperiodes van 10 maande en langer word die gebruik van bevriesing en verkoeling aanbeveel as die mees geskikte preserveringstegnieke. AI drie verpakkingsmateriale kan gebruik word vir die verpakking en opberging van gevriesdroogde Kepikorrels vir 'n tydperk van een maand. Indien 'n opbergingsperiode van twee maande of langer verlang word, word die OPET-verpakkingsmateriaal aanbeveel vir die suksesvolle behoud van die fermentasie-aktiwiteit van die Kepikorrels.