Kefir is a traditional fermented milk originating many centuries ago in the Caucasian mountains. It is produced by fermentative activity of ‘kefir grains’ consisting mainly of lactococci, lactobacilli and yeasts in a protein–polysaccharide matrix. The grains contain a relatively stable and specific balance of microorganisms which exist in a complex symbiotic relationship. The grains grow in the process of kefir making only from pre-existing grains (Saloff-Coste, 1996). When kefir grains are allowed to grow in milk, microorganisms are shed from the grains into milk where they continue to multiply with the production of acid, flavour and physicochemical changes.

The traditional method of kefir making is currently by adding kefir grains directly as starter to milk that has been pasteurized and cooled to 20–25°C. After a period of fermentation lasting ∼24 h, the grains are removed by filtration and the beverage is ready for consumption (Saloff-Coste, 1996). Kefir from which the grains have been removed may be used as starter. However, this fermented milk cannot be used for subsequent inoculations to make an acceptable product, because the original balance of microorganisms has been disrupted (Kroger, 1993).

The complex microbiological composition of kefir grains explains why it is difficult to obtain starter with the optimal and constant composition necessary for a regular production of kefir of standard quality (Koroleva, 1988a). Studies have been undertaken to establish cultivation conditions[ratio ]grain[ratio ]milk ratio, cultivation temperature, period of time and conditions prior to separation of grains from the fermented milk, shaking conditions for agitation of milk with the grains in the course of fermentation, washing of kefir grains and so on. All these factors influence the microflora of the kefir starter and fermented milk. There are no rules about household manufacture of kefir. Different reports indicate a wide range of grain[ratio ]milk ratios for kefir making. Bottazzi & Bianchi (1980), Marshall & Cole (1985), Merin & Rosenthal (1986), Mann (1989), Hosono et al. (1990) and Kroger (1993) employed 20–50 g/l while Koroleva (1988a) employed 20–100 g kefir grain/l and Marshall et al. (1984) and Neve (1992) 50–100 g/l. Rea et al. (1996) used 1 g kefir grain/l as starter and 200 g starter in the form of kefir grains is recommended by Hansen for the fermentation of 1 l milk (Marshall & Cole, 1985). A critical control point in kefir manufacture to obtain a product with constant quality is the standardization of the kefir grain[ratio ]milk ratio. Koroleva (1988b) claimed that it is better to use kefir grains as starter for kefir production and, at the same time, to decrease the amount of inoculum.

The purpose of this study was to evaluate the effects of changes in the kefir grain[ratio ]milk ratio (quantity of kefir grains inoculated into the milk) on microflora composition, acidity, apparent viscosity and carbon dioxide content of fermented milk.