Generally, Streptococcus thermophilus ferments sucrose, whereas Lactobacillus bulgaricus fails to utilize this disaccharide. When both sucrose and lactose were added to a basal medium, S. thermophilus fermented both carbon sources, produced sufficient acid to change the color of an acid-base indicator (bromcresol purple), and hence formed yellow colonies. On the same medium, most L. bulgaricus strains grew more slowly, produced less acid, and yielded white colonies. Acid diffusion around the S. thermophilus colonies was localized by incorporation of CaCO3 into the medium. To test the efficacy of this medium when known strains of starters are used, the effect of freezing with liquid nitrogen on mixed cultures of S. thermophilus and L. bulgaricus was studied.

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Psychrotrophic bacteria were isolated from 227 pasteurized milk samples which had a shelf life in excess of 20 days at 7.2 C. Of 700 cultures isolated, 135 were resistant to heating at 72 C for 16 sec and were able to re-establish growth at 7.2 C. Thirty-five cultures, representing 15 different types were subjected to detailed examination to determine their actions on refrigerated milk, growth temperatures, thermal resistance at various temperatures, and their identities. The spore-forming genus Bacillus occured most frequently. The non-sporing types were assigned to the genera Arthrobacter, Microbacterium, Streptococcus, and Corynebacterium.

Eight lots of Cheddar cheese were manufactured with two strains each of Streptococcus faecalis and Streptococcus durans and subjected to combinations of two early cooling treatments (air vs. brine cooling) and two curing temperatures (7.2 and 12.8 C). The enterococcus cultures were used as supplemental starters in combination with a commercial lactic culture. These cheeses were analyzed for microbiological growth and survival, proteolysis, lactic acid development, free fatty acid appearance, and citric acid utilization—each being compared with a control cheese made without enterococci. Results were presented in three previous articles. This series is concluded with the results of organoleptic ana1ysis of the cheeses. Cheeses made with S. faecalis were either comparable to or less desirable than their respective control cheeses. Those made with S. durans, however, were in all instances more desirable than their controls. Cheeses cured at 7.2 C were always given the better scores, but there was no statistically significant difference between air- and brine-cooled cheeses.

An agar medium containing arginine and calcium citrate as specific substrates, diffusible (K 2 HPO 4 ) and undiffusible (CaCO 3 ) buffer systems, and bromocresol purple as the p H indicator was developed to differentiate among lactic streptococci in pure and mixed cultures. Milk was added as the sole source of carbohydrate (lactose) and to provide growth-stimulating factors. Production of acid from lactose caused developing bacterial colonies to seem yellow. Subsequent arginine utilization by Streptococcus lactis and S. diacetilactis liberated ammonia, resulting in a localized p H shift back toward neutrality and a return of the original purple indicator hue. The effects of production of acid from lactose and ammonia were fixed around individual colonies by the buffering capacity of CaCO 3 . After 36 hr at 32 C in a candle oats jar, colonies of S. cremoris were yellow, whereas colonies of S. lactis and S. diacetilactis were white. S. diacetilactis , on further incubation, utilized suspended calcium citrate, and, after 6 days, the citrate-degrading colonies exhibited clear zoning against a turbid background, making them easily distinguishable from the colonies of the other two species. The medium proved suitable for quantitative differential enumeration when compared with another widely used general agar medium for lactic streptococci.