A survey was undertaken at a whole milk powder manufacturing plant to determine the origin and the rate of spore formation of thermophilic bacteria. Spores were generally not detected (< 10 cfu/mL) in either the pasteurization process or the pasteurized milk directed into the powder plant. The predominant sites of spore formation were the preheater plate heat exchanger and the evaporator. Spores began to be detected approximately 9 h into an 18‐h manufacturing run. Spore isolates were identified as Anoxybacillus flavithermus and Geobacillus species. A. flavithermus predominated in the preheat section, whereas a mix of both organisms was present in the later manufacturing stages.

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Six Lactobacillus acidophilus, 5 Bifidobacterium, and 6 Streptococcus thermophilus strains were studied for characteristics that are important to activity and stability in unfermented fluid milk products. Speciation, strain relatedness, frozen concentrate stability, bile sensitivity, and lactase activity were evaluated. The microbiological stability of a culture-containing fluid milk product was also determined. Two of the bifidobacteria cultures contained > 1 strain. Some strains were shown to be closely related or identical by pulsed-field gel electrophoresis of fragmented chromosomal DNA. Selective media that distinguished among all 3 added genera were identified. All lactobacilli and most of the bifidobacteria were resistant to bile concentrations varying from 1 to 3%, and all streptococci were sensitive to bile. Lactase activities were highest for S. thermophilus strains, supporting use of this species in fluid milk and dairy products to aid in the digestion of lactose by consumers. The experimental product evaluated in this study contained 10(7) cfu/ml of both L. Acidophilus and Bifidobacterium spp. and 5 x 10(7) cfu/ml of S. thermophilus. Lactic, but not psychrotrophic, populations were fairly stable during storage. The results suggest that milk formulated with high concentrations of three different genera of probiotic bacteria can be manufactured with commercial strains.

Aims:  To examine the rate and the extent of spore formation in Anoxybacillus flavithermus biofilms and to test the effect of one key variable – temperature – on spore formation. Methods and Results:  A continuous flow laboratory reactor was used to grow biofilms of the typical dairy thermophile A. flavithermus (strain CM) in skim milk. The reactor was inoculated with either a washed culture or a spore suspension of A. flavithermus CM, and was run over an 8·5 h period at three different temperatures of 48, 55 and 60°C. Change in impedance was used to determine the cell numbers in the milk and on the surface of the stainless steel reactor tubes. The biofilm developed at all three temperatures within 6–8 h. Spores formed at 55 and 60°C and amounted to approx. 10–50% of the biofilm. No spores formed at 48°C. Conclusions:  The results suggest that both biofilm formation and spore formation of A. flavithermus can occur very rapidly and simultaneously. In addition, temperature variation has a considerable effect on the formation of spores. Significance and Impact of the Study:  This information will provide direction for developing improved ways in which to manipulate conditions in milk powder manufacturing plants to control biofilms and spores of A. flavithermus.

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