Equol, a derivative of daidzein produced by enterobacteria, has greater activity as a phyto-oestrogen compared with daidzein. Difructose anhydride III (DFAIII) is a newly manufactured non-digestible disaccharide with unique fermentation properties. The present study evaluated the prebiotic effects of DFAIII on equol production and on plasma cholesterol concentrations related to the changes in equol production. We compared plasma equol concentrations at 10.00 and 18.00 hours and faecal isoflavone excretion in three groups of seven rats (male Wistar-ST strain, 6 weeks old) fed a basal diet or a DFAIII or fructooligosaccharide (15/g diet) diet containing 1g soya isoflavone/g diet for 20d. Equol concentrations in the DFAIII group were higher than in the control and fructooligosaccharides groups, especially in the later phase of the light period (18.00 hours) throughout the experiment. Daizein and genistein concentrations did not change between the diet groups. The faecal ratios of equol:daidzein were very high in all groups, but the ratios were higher in the DFAIII group than the control and fructooligosaccharide groups on day 3, and this tendency continued throughout the experiment. On day 20, the plasma total cholesterol concentration was lowest in the DFAIII group. Additionally, the cholesterol concentrations were inversely correlated to plasma equol concentration in all the rats. In conclusion, DFAIII efficiently enhanced plasma equol concentrations, which may be associated with an increase in equol production and a decrease in equol degradation by enterobacteria. Higher plasma equol concentrations may contribute to the hypocholesterolaemic effect of DFAIII feeding.

This study demonstrates that feeding difructose anhydride III (DFAIII) improves bone strength and femoral mineral concentrations in a rat model of oestrogen deficiency. We showed the relationship between Ca, Mg and P absorption and bone characteristics in rats. Two groups of female Sprague-Dawley rats (6 weeks old) underwent bilateral ovariectomy (ovariectomized rats, OVX rats) or bilateral laparotomy (sham rats). At 10 weeks old, OVX and sham rats were divided into three subgroups and fed a control, 1·5 % DFAIII or 3 % DFAIII diet for 8 weeks, respectively. Ca but not Mg absorption rates were lowered by ovariectomy; however, ingestion of the 1·5 % and 3 % DFAIII diets similarly restored the reduced Ca absorption in OVX rats at 4 and 8 weeks after feeding of the test diets. DFAIII increased Mg absorption dose-dependently in sham and OVX rats. The bone strength, femoral Ca and Mg concentrations, and distal bone mineral density in the 3 % DFAIII group were higher than those in the control group in OVX rats. The absorption rates of Ca and Mg were significantly correlated with femoral Ca and Mg concentrations and strength, which suggests that increasing both Ca and Mg absorption improves bone characteristics in OVX rats. There were no differences in any of the variables in the femur between the 1·5 % and 3 % DFAIII groups in OVX rats. In conclusion, feeding of a low dose of DFAIII increased intestinal Ca and Mg absorption, and the promotive effect of DFAIII persisted for over 8 weeks. This effect was associated with prevention of ovariectomy-induced osteopenia.