Field observations and interviews were conducted in fisherfolk households of Cordova Municipality, Mactan Island, Province of Cebu, Philippines to elucidate their usage of milk and milk products, the timing to adopt milk and milk products into their livelihoods, and then to verify the spread and acceptance of five forms of milk culture into the non-milk cultural sphere. The milk and milk products were never used in traditional main diets such as seafoods, but consumed for breakfast with bread, biscuits or rice and for a snack as sweets. The milk and milk products mainly spread to the Philippines in colonial occupation periods from Spain, Japan and America. It was concluded that the milk and milk products were processed into very sweet products in the hot and humid environment of the Philippines, but did not influence the traditional main diets basically consisting of seafoods. They comprised four of the original five forms of milk culture, ‘supplementary nutrition’, ‘favorite’, ‘fusion with rice’ and ‘western-style food culture’ into the Philippines. These same trends are also found in other non-milk cultural spheres such as Southeast Asian countries and Japan. It is thought that these characteristics are the first stage in spreading and accepting milk culture into non-milk cultural spheres in general.

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　Hydrogen peroxide is considered one of the important factors that decrease the viability of the probiotic Bifidobacterium in yogurt. In the present study, we investigated the effect of hydrogen peroxide produced by yogurt starters on the viability of Bifidobacterium. We first evaluated the hydrogen peroxide productivity of 11 yogurt starter strains (Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus). The amount of hydrogen peroxide produced by L. bulgaricus was five times higher than that produced by S. thermophilus cultivated in 10% (w/v) reconstituted skim milk at 37℃ for 16 h. Then, hydrogen peroxide production by L. bulgaricus under anaerobic conditions was 65% lower than that under aerobic conditions. In addition, generation of hydrogen peroxide was associated with bacterial cell counts of L. bulgaricus rather than those of S. thermophilus in a lactate buffer. These results indicate that the accumulation of hydrogen peroxide in yogurt is attributed to oxygen metabolism of L. bulgaricus. Reduction in inoculated cell number of L. bulgaricus decreased the accumulation of hydrogen peroxide and significantly improved the viability of Bifidobacterium adolescentis KH 96 in yogurt during storage.

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　The purpose of this study is to investigate lactic acid bacterial distribution in raw milk of Wagyu cattle, and to evaluate a tolerance of the lactic acid bacteria against simulated digestive juices. Isolates from raw milk were identified as Lactobacillus reuteri, Lactobacillus plantarum, Lactococcus garvieae, Lactococcus lactis, Enterococcus hirae, Enterococcus faecalis, Enterococcus saccharolyticus, Weissella thailandensis and Pediococcus pentosaceus based on their physiological and biochemical characteristics as well as by 16S rDNA sequence analysis. Lb. reuteri PUHM1004 and E. hirae PUHM1011 exhibited more than 50% of survival rate after 6 hours incubation in MRS broth supplemented with 0.3% oxgall. These strains survived after 3 hours in simulated gastric juice adjusted pH 2.0, and grew in a simulated intestinal juice. These strains could be used for probiotic food and feed.

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　Constipation is a serious problem to decrease the quality of life following socioeconomic loss. Milk whey culture with Propionibacterium freudenreichii ET-3 (milk whey culture) has been reported to improve the constipation by enhancing the growth of Bifidobacterium in clinical study. However, the effect of milk whey culture on the colonic motility has not been clarified. Therefore, this study aimed to clarify this point using the rat colon in vitro. To clarify the regional specificity, longitudinal specimens of the proximal, middle and distal colon were used. After obtaining the constant spontaneous contraction, milk whey culture was added in a concentration-dependent manner. The contractile frequency and force were used as end-point. Milk whey culture significantly enhanced the contractile frequency of the distal colon in a concentration-dependent manner, but not the contractile force. On the contrary, in the proximal and middle colon, milk whey culture did not show any significant effect in both contractile frequency and force. Milk whey culture contains 1,4-dihydroxy-2-naphthoic acid (DHNA) enhancing the growth of Bifidobacterim. DHNA did not influence the contractile frequency and force. Also propionic acid and acetic acid contained in milk whey culture did not show any significant effect. Above results show that milk whey culture may relief the constipation by exclusively enhancing the spontaneous contraction of the distal colon, which plays an important role for defecation, although active substance was not clarified. No effect on the contractile force may be a merit, because the enhancement of contractile force may lead to the abdominal pain.

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　Immune-enhancing enteral formulas are used in patients who have undergone surgical treatment or trauma in order to reduce the risk of infectious disease and duration of hospitalization. The formulas are generally fortified with nutrients such as arginine, glutamine, n-3 fatty acids, nucleic acids, and trace elements. In this study, we developed a novel immune-enhancing formula fortified with a multi-functional protein, bovine lactoferrin (bLF), as well as immune-enhancers such as arginine, glycine, glutamine, and nucleic acids. We then assessed the stability of bLF in the formula during long-term storage and its health benefits as an immune-enhancer. Although bLF is extremely heat-labile during the sterilization process of enteral formulas, denaturation can be reduced considerably by sterilizing bLF using direct-ultra-high-temperature sterilization under acidic conditions (pH 2.7). The bLF latex agglutination test showed no loss of bLF during a 9-month period at 25℃ and 37℃, indicating its stability. Although bLF in the formula was highly iron-saturated and did not exhibit a bacteriostatic effect on Escherichia coli, the peptic hydrolysate of the bLF, simulating gastric digestion, released the peptide of lactoferricin and exhibited a strong bactericidal effect. Mice fed with this formula for 90 days displayed longer ileal villi, although this difference did not reach statistical significance, and altered intestinal microbiota composition with a significant increase in Lactobacillales detected. Thus, this formula is distributable at ambient temperature for months and may be useful to prevent infectious diseases by altering microbiota composition to provide a better enteral environment.

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