No abstract

Effects of water activity and storage temperature on survival of bifidobacteria in powder form were investigated and kinetic analyses were performed to reveal characteristics of the stability. A significant positive correlation was observed between water activity and natural logarithm of the inactivation rate constant of bifidobacteria powder, indicating that higher water activity induced lower stability of bifidobacteria in powder form. Also, higher temperature condition induced lower survival rate, which was supported by that the stability was followed the Arrhenius theory. These findings constructed a prediction model for bifidobacteria survival in powder form.

The effects were examined of a pepsin hydrolysate of bovine lactoferrin on the proliferation of murine splenocytes. The hydrolysate enhanced [3H]thymidine uptake by splenocytes, but undigested bovine lactoferrin exerted an inhibitory effect. The hydrolysate had the ability to inhibit the blastogenesis that was induced by mitogens such as concanavalin A, phytohemagglutinin, and lipopolysaccharide; inhibition was similar to that with undigested lactoferrin. These results suggested that the hydrolysate contained both immunostimulatory and immunoinhibitory peptides. The stimulatory effect of the hydrolysate in the absence of mitogens was then explored in more detail using nonadherent splenocytes. The proliferative response of splenocytes to the hydrolysate was much greater in the fraction that was enriched with B cells than in the fraction that was enriched with T cells. The hydrolysate did not affect thymocyte proliferation. These data indicated that the adherent cells resembling macrophages and found among the splenocytes were not the target cells of the hydrolysate. The stimulatory effect of the hydrolysate was due to the activation of B cells by the hydrolysate and enhanced immunoglobulin production by splenocytes. Because the hydrolysate also enhanced the proliferation and immunoglobulin A production of Peyer's Patch cells, the immunostimulatory effect of the hydrolysate in vivo was examined using mice that had been orally immunized with cholera toxin. The concentrations of immunoglobulin A conjugated against cholera toxin in bile and in the intestinal contents of mice fed liquid diets containing 1% (wt/vol) lactoferrin hydrolysate were greater than those of mice fed control diets. This result suggested that the use of the lactoferrin hydrolysate is beneficial to enhance mucosal immunity.

Two stability tests of bifidobacteria in powdered formula were conducted. In a strain comparison test, three kinds of bifidobacterial powders; Bifidobacterium longum BB536, Bifidobacterium breve M-16V and Bifidobacterium infantis M-63 powders, admixed in follow-up formula were used. In a commercial product evaluation, powdered formulas for toddlers containing bifidobacteria sold in the Indonesian market were analysed. When the inactivation rate constant of each sample, which was used as an index of the loss rate, was determined from the stability tests, B. longum was the most stable strain. The mean inactivation rate constant of commercial products was significantly lower than those obtained in strain comparison, although the same strains (B. longum BB536 and B. breve M-16V) were used. A possible reason was the lower water activity of commercial products compared to the follow-up formula. Also, higher storage temperature yielded lower stability in all strains or samples, which obeys the Arrhenius theory.

Feeding of bovine milk to mice induced a high incidence of bacterial translocation from the intestines to the mesenteric lymph nodes, and the bacteria involved were mainly members of the family Enterobacteriaceae. Supplementation of the milk diet with bovine lactoferrin or a pepsin-generated hydrolysate of bovine lactoferrin resulted in significant suppression of bacterial translocation. Our findings suggest that this ability of lactoferrin to inhibit bacterial translocation may be due to its suppression of bacterial overgrowth in the guts of milk-fed mice.