Stanley E. Gilliland

The reducing ability and antioxidative activity of some species of Lactobacillus were compared under in vitro conditions. Cultures of Lactobacillus delbrueckii ssp. lactis, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus, and Lactobacillus casei were grown at 37 degrees C in de Man, Rogosa, Sharpe (MRS) broth supplemented with 0.5% 2,3,5 triphenyl tetrazolium chloride (TTC) to evaluate reducing activity. Reduced TTC was extracted from the cultures with acetone, and the intensity of the red color measured colorimetrically at 485 nm was an indication of reducing activity. The lactobacilli varied significantly in relative ability to reduce TTC when grown in MRS broth for 15 h. The relative amounts of growth as indicated by pH values at 18 h appeared to influence the amount of reduction. Antioxidative activity was evaluated by the ability of the whole cells or the cell-free extracts from cultures to protect a protein from being attacked by free radicals. These analyses were performed using the oxygen radical absorbance capacity method. All cultures tested exhibited some degree of antioxidative activity. Among the treatments, the cell-free extracts from cells grown in MRS broth exhibited significantly higher values than did whole cells. There was no apparent relationship between the reducing and antioxidative activities of the cultures evaluated. The results from this study show that these cultures can provide a source of dietary antioxidants. Furthermore, selection of cultures that produce antioxidants as starters could provide yet another health or nutritional benefit from cultured or culture-containing dairy products.

Cells of Lactobacillus delbrueckii ssp. lactis RM2-5 were added to cottage cheese stored at 7 degrees C in different amounts to determine if they would inhibit the growth of Pseudomonas fluorescens, also inoculated into the cheese samples. In addition, experiments were conducted in which no spoilage organisms were added to determine the effect of the lactobacilli on the natural background flora in the cottage cheese. For most experiments, as the numbers of lactobacilli increased, the numbers of spoilage organisms were lower than in the control on any given day of storage. In cheese inoculated with P. fluorescens, the numbers of spoilage organisms in the control had increased 5 log cycles by d 7, whereas the treatment containing the highest level of L. delbrueckii ssp. lactis RM2-5 (1.0 x 10(9) cfu/g) had not, and did not, increase over the course of the 21-d study. In the experiments where no spoilage organisms were added, lactobacilli significantly retarded the growth of gram-negative bacteria in the cheese. However, in these experiments, mold growth on the samples became a limiting factor during extended storage. The results of these experiments indicate that lactobacilli could be effective at helping control gram-negative spoilage bacteria in cottage cheese, thus potentially extending its shelf life.

Methanol, acetaldehyde, ethanol, and hexanal were the 4 major volatiles detected in unfermented soymilk. Eight of the cultures of lactobacilli or streptococci completely eliminated hexanal in the soymilk during fermentation. However, there were considerable variations in the effects of the cultures on the other 3 compounds. All 8 caused significant reduction in levels of methanol. Streptococcus thermophilus OSU‐2 was the only culture that significantly lowered the concentration of ethanol in the soymilk. All except Lactobacillus acidophilus C19 and Lactobacillus casei E5 significantly lowered the level of acetaldehyde. Of the cultures tested, L. acidophilus L1 offered the best potential for producing fermented soymilk with an improved volatile profile.

Growth of some probiotic bacteria was significantly improved in milk supplemented with whey protein hydrolysate (WPH). However, WPH had no effect on the growth of Lactobacillus delbrueckii ssp. bulgaricus 18, L. delbrueckii ssp. bulgaricus 10442, and Streptococcus thermophilus 1. When the probiotic bacteria were grown in combination with different yogurt cultures in milk, WPH caused significant increases in growth of Bifidobacterium longum S9, L. acidophilus O16, and L. acidophilus L‐1. However, by day 28 of refrigerated storage, the populations of the probiotic cultures that had been grown in samples supplemented with WPH were similar or below those in the control samples.

Binding of sodium cholate, sodium taurocholate, and sodium glycocholate by guar gum, soluble oat fiber, xanthan gum, and inulin was studied. All soluble fibers were able to bind sodium cholate, sodium taurocholate, and sodium glycocholate from a mixture of the three, but when tested individually, the fibers bound little or no sodium cholate. In general, the fibers bound higher concentrations of conjugated bile salts than they did free bile salts. Soluble oat fiber, which bound as much or more sodium cholate and sodium glycocholate as did the other 3 fibers, was tested for its effect on deconjugation of glycocholate by Lactobacillus acidophilus and Lactobacillus casei. The presence of the fiber significantly increased (P < 0.05) deconjugation by all cultures.

Raw vegetables inoculated with selected pathogenic bacteria were treated with a strain of Lactobacillus delbrueckii subsp. lactis, which was selected for its ability to produce hydrogen peroxide at refrigerated temperatures. The vegetables inoculated included broccoli, cabbage, carrots, and lettuce. Each vegetable was rinsed, chopped, and stored under conditions similar to those used for ready-to-eat vegetables sold at retail. Portions of each vegetable were separately inoculated with one of two pathogenic bacteria, Escherichia coli O157:H7 or Listeria monocytogenes. Prior to packaging, one portion of each inoculated vegetable was treated with a cell suspension of the selected strain of L. delbrueckii subsp. lactis. The vegetables were stored at 7°C for 6 days. The populations of pathogens and lactobacilli on each sample were enumerated on storage days 0, 3, and 6. Although populations of L. delbrueckii subsp. lactis remained at high levels during storage, there was no noticeable antagonistic action again...

A gas chromatographic procedure was used for analysis of conjugated linoleic acid (CLA) isomers cis-9, trans-11-octadecadienoic; trans-10, cis-12 octadecadienoic; and trans-9, trans-11-octadecadienoic (c9t11, t10c12, t9t11) produced by lactobacilli. Four different cultures, two strains each of Lactobacillus acidophilus and Lactobacillus casei were tested for their ability to produce CLA from free linoleic acid in MRS broth supplemented with linoleic acid. Different concentrations of linoleic acid (0, 0.05, 0.1, 0.2 and 0.5 mg/ml) were added to MRS broth, inoculated with the lactobacilli, and incubated at 37 degrees C. Viable counts and amounts of individual isomers of CLA (c9t11, t10c12, t9t11) were measured at 0, 24, 48, and 72 h. All the cultures were able to produce free CLA in media supplemented with linoleic acid. Maximum production of CLA (80.14 to 131.63 microg/ml) was observed at 24 h of incubation in broth containing 0.02% of free linoleic acid. No significant (P > 0.05) increases in total CLA levels were observed after 24 h of incubation. The ability of the cultures to produce CLA in skim milk supplemented with 0.02% free linoleic acid also was studied. In this medium, the total amounts of free CLA after 24 h of incubation ranged from 54.31 to 116.53 microg/ml. The use of lactic acid bacteria able to form free CLA in cultured dairy products may have potential health or nutritional benefits. Free CLA in the products likely would be more readily available for absorption from the digestive tract than if it were incorporated into the cells of the starter culture.

Cells of Lactobacillus delbrueckii subsp. lactis RM2-5 were added to various meat model systems that had been inoculated with Escherichia coli O157:H7 or Salmonella Typhimurium to determine whether...

Several strains of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus, which produced exocellular polysaccharides (EPS), varied in the amount produced. The streptococci tended to produce the most EPS per milliliter of culture; however, when compared on the basis of amounts per 10(7) cfu, the lactobacilli produced the most. Lactobacillus delbrueckii ssp. bulgaricus strains Lb-18 and Lb-10442 and S. thermophilus St-143 produced significantly larger amounts per 107 cfu than did other strains tested. These three cultures plus two strains of the streptococci that produced the greatest amounts of EPS per ml of culture were tested for the ability to bind bile acids from laboratory media. The two cultures of L. delbrueckii ssp. bulgaricus (Lb-18 and Lb-10442) bound significantly higher amounts of cholic acid than did the three strains of streptococci. These two cultures of lactobacilli bound up to 15.3% of the cholic acid present in laboratory media, up to 452 microg/mg of EPS and 2.9 microg/10(7) cfu. None of the cultures tested in this study were able to bind the conjugated bile acid, glycocholic acid.