1. General Nutritional Requirement
2. Trials to Establish Bifidus-Flora
Administration of living bifidobacterial preparations
Improvement of powdered milk
Search for bifidus-factors
Bifidus-factor I
Bifidus-factor II
Lactulose
Bifidus-factor in carrot root
Other factors
Search for sugar sources
Selectivity
Efficiency
Further screening
3. Proposal for Bifidobacteriology

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We studied the effects of administration of TOS, a new growth factor derived from lactose for Bifidobacterium, and Bifidobacterium breve 4006 on the fecal flora of normal subjects. All of the Bifidobacterium species tested, eight reference strains and B. breve 4006 were capable of fermenting TOS in vitro, while others, 2 Bacteroides strains and 4 Lactobacillus and Enterobacteriaceae strains, showed an appreciable growth among 55 cultures tested. It was evident that TOS is not intestinally absorbed by the recipient subjects, from hydrogen breath test. In vivo, TOS (3g or 10g/day) was observed to promote the growth of both administered B. breve 4006 and resident Bifidobacterium strains. Simultaneous administration of B. breve 4006 and TOS caused the suppression of gram negative anaerobes and aerobes, Bacteroidaceae and Enterobacteriaceae, and the reduction of fecal ammonia and urinary indican excretion. It is concluded that TOS is a typical bifidus factor.

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This review deals with the effects of composition of infant formulas on the intestinal microflora of infants. The addition of bifidus factor, such as lactulose and mucin, works effectively to stimulate the growth of bifidobacteria, lower pH values and raise the lysozyme activity in the feces of bottle-fed infants. Also, low buffering capacity resultant from reducing the protein and phosphorus content in the formula contributes to the predominance of bifidobacteria. These facts indicate that the addition of bifidus factor and low buffering capacity play a significant role to resemble the fecal microflora of bottle-fed infants to that of breast-fed infants through the enhancing multiplication of bifidobacteria, though the ratio of bifidobacteria count to total anaerobic bacteria count in bottle-fed infants is still lower than that in breast-fed infants.

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In order to obtain current information about the intestinal flora of bottle-fed infants, feces from 30 breast-fed infants and 43 bottle-fed infants, including 3 infants of low birth weights, were examined by the method of Mitsuoka. The frequency of occurrence and total number of bifidobacteria in bottle-fed infants were nearly equal to those of breast-fed infants. The number of enterobacteriaceae, streptococci, and anaerobes other than bifidobacteria were significantly higher in bottle -fed infants than in breastfed infants. As for Bifidobacterium species, there was little difference between bottlefed and breast-fed infants in average counts of all the species detected, but the frequency of occurrence of Bifidobacterium bifidum in bottle-fed infants was less than in breast-fed infants. The appearance of bifidobacteria in low birth weight bottle -fed infants was more delayed than that in normal bottle-fed infants, and their species composition was also simple, consisting of two species of bifidobacteria: B. breve plus B. bifidum, or B. breve plus B. adolescentis.

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Effect of dried viable bifidobacteria preparations (108 B. thermophilum/g and 109 B. pseudolongum/g) on intestinal flora and clinical symptoms of scouring animals were investigated by oral administration to dogs, calves and pigs. Disturbances of bacterial flora while scouring were only seen in some limited bacterial species. In scouring animals, the population and/or incidence of bifidobacteria and lactobacilli obviously decreased while counts of enterobacteriaceae increased. Appearance of hemolytic E. coli and streptococci, and higher counts of Clostridium perfringens in scouring dogs were characteristic. Oral administration of the bifidobacteria preparation seemed to reinforce recovery of normal intestinal flora and alleviate clinical symptoms of scouring animals. Prophylactic effects of the preparation were observed in preventing diarrhea in weaned puppies, early weaned bull calves and suckling piglets.

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