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Background and Aim: We previously reported the mitigation effects of difructose anhydride III (DFA III) on mycotoxins, such as zearalenon and sterigmatocystin, based on the urinary concentrations of these molecules in calves. This study was aimed at evaluating the effects of dietary supplementation of DFA III and the fermented status of DFA III in the intestine by comparing serum levels of short-chain fatty acid (SCFAs) in DFA III-supplemented cattle with those in non-supplemented control cattle. Materials and Methods: Serum SCFA concentrations were measured in 30 Japanese Black heifers, aged 9–10 months, from two herds, using gas chromatography on days 0 (before DFA III supplementation), 9, and 14 after DFA III supplementation. Results: A notably different trend was observed for isobutyric acid and enanthic acid, which may reflect the different fermentation status of supplementary DFA III in the intestine. Our results indicate the possibility that this trend observed in the intestinal tract following DFA III administration is associated with changes in the environment of intestinal bacterial flora, which may partially reflect the effects of DFA III supplementation on cattle. Conclusion: Difructose anhydride III supplementation for at least 2 weeks affects the trend of blood SCFA concentrations in cattle. Our results provide evidence supporting the effects of DFA III on the intestinal environment and intestinal barrier function.

Background and Aim: We previously reported the mitigation effects of difructose anhydride III (DFA III) on mycotoxins, such as zearalenon and sterigmatocystin, based on the urinary concentrations of these molecules in calves. This study was aimed at evaluating the effects of dietary supplementation of DFA III and the fermented status of DFA III in the intestine by comparing serum levels of short-chain fatty acid (SCFAs) in DFA III-supplemented cattle with those in non-supplemented control cattle. Materials and Methods: Serum SCFA concentrations were measured in 30 Japanese Black heifers, aged 9-10 months, from two herds, using gas chromatography on days 0 (before DFA III supplementation), 9, and 14 after DFA III supplementation. Results: A notably different trend was observed for isobutyric acid and enanthic acid, which may reflect the different fermentation status of supplementary DFA III in the intestine. Our results indicate the possibility that this trend observed in the intestinal tract following DFA III administration is associated with changes in the environment of intestinal bacterial flora, which may partially reflect the effects of DFA III supplementation on cattle. Conclusion: Difructose anhydride III supplementation for at least 2 weeks affects the trend of blood SCFA concentrations in cattle. Our results provide evidence supporting the effects of DFA III on the intestinal environment and intestinal barrier function. [ABSTRACT FROM AUTHOR]

Production and applications of difructose anhydride III (DFA-III) have attracted considerable attention because of its versatile physiological functions. Recently, large-scale production of DFA-III has been continuously explored, which opens a horizon for applications in the food and pharmaceutical industries. This review updates recent advances involving DFA-III, including: biosynthetic strategies, purification, and large-scale production of DFA-III; physiological functions of DFA-III and related mechanisms; DFA-III safety evaluations; present applications in food systems, existing problems, and further research prospects. Currently, enzymatic synthesis of DFA-III has been conducted both industrially and in academic research. Two biosynthetic strategies for DFA-III production are summarized: single- and double enzyme-mediated. DFA-III purification is achieved via yeast fermentation. Enzyme membrane bioreactors have been applied to meet the large-scale production demands for DFA-III. In addition, the primary physiological functions of DFA-III and their underlying mechanisms have been proposed. However, current applications of DFA-III are limited. Further research regarding DFA-III should focus on commercial production and purification, comprehensive study of physiological properties, extensive investigation of large-scale human experiments, and expansion of industrial applications. It is worthy to dig deep into potential application and commercial value of DFA-III. [ABSTRACT FROM AUTHOR]

The functional Difructose anhydride III (DFA-III) lacks reported utilization by special probiotics of Bifidobacterium and Lactobacillus. DFA-III hydrolase (DFA-IIIase), converting DFA-III to inulobiose, is a critical enzyme for the metabolism of DFA-III, which stands for the utilization ability of DFA-III by microorganisms. Hence, the research identified six potential DFA-IIIases from Bifidobacterium and Lactobacillus species, suggesting that DFA-III has the potential to proliferate these bacteria. Notably, the DFA-IIIase from Bifidobacterium adolescentis belonging to the human intestinal microbe exhibits a hydrolysis rate of up to 67 % for DFA-III, which is the highest among the reported DFA-IIIases to date. When DFA-IIIases were applied to Jerusalem artichokes, DFA-III, inulobiose, and fructo-oligosaccharides were significantly produced. The in vitro work indicates that Bifidobacterium and Lactobacillus have the potential ability to utilize DFA-III by DFA-IIIases. Moreover, the first application of DFA-IIIase in Jerusalem artichokes provides valuable insights into comprehensive strategies for utilizing high-inulin agricultural products., Competing Interests: Declaration of competing interest The authors declare that no competing interests exist., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The relative available energy (RAE) based on the fermentability of breath hydrogen excreted by gut microbiota was assessed using a method accepted by the Japanese Society for Dietary Fiber Research. The purpose of this study was to evaluate RAE of difructose anhydride III (DFA III) and maltobionic acid (MA). In this study, which had a within-subject, repeated measures design, 11 healthy female participants ingested 5 g of DFA III, MA, and fructooligosaccharide, which has RAE of 2 kcal/g. Breath hydrogen was collected 12 times up to 14 h after meal consumption and measured using gas chromatography, and the area under the curves (AUCs) was calculated 0–14 h after ingestion. Compared with the AUCs of breath hydrogen excretion related to the ingested fructooligosaccharide for 14 h, those of DFA III and MA were 40.5% and 23.1%, respectively. RAE was 0.81 kcal/g for DFA III and 0.46 kcal/g for MA, respectively.

The relative available energy (RAE) based on the fermentability of breath hydrogen excreted by gut microbiota was assessed using a method accepted by the Japanese Society for Dietary Fiber Research. The purpose of this study was to evaluate RAE of difructose anhydride III (DFA III) and maltobionic acid (MA). In this study, which had a within-subject, repeated measures design, 11 healthy female participants ingested 5 g of DFA III, MA, and fructooligosaccharide, which has RAE of 2 kcal/g. Breath hydrogen was collected 12 times up to 14 h after meal consumption and measured using gas chromatography, and the area under the curves (AUCs) was calculated 0–14 h after ingestion. Compared with the AUCs of breath hydrogen excretion related to the ingested fructooligosaccharide for 14 h, those of DFA III and MA were 40.5% and 23.1%, respectively. RAE was 0.81 kcal/g for DFA III and 0.46 kcal/g for MA, respectively. [ABSTRACT FROM AUTHOR]

• A recombinant Saccharomyces cerevisiae hyper-secreting inulin fructotransferase (161.6 U/mL) was developed. • The strain produced 77.5 g of DFA III from 200 g of Jerusalem artichoke tuber powder. • By additional culture of Kluyveromyces marxianus , the DFA III content based on carbohydrates was increased up to 95%. • This is the first direct production of DFA III from crude JAP by microbial fermentation. Difructose anhydride III (DFA III), a functional sweetener, is enzymatically produced from inulin by inulin fructotransferase (IFTase; EC 4.2.2.18). Here, a recombinant yeast strain hyper-secreting IFTase from Arthrobacter aurescens was developed by employing the translational fusion partner technique, and the recombinant yeast produced 77.5 g (64.6% yield) of DFA III from 200 g of crude extract of Jerusalem artichoke tuber powder containing 120 g of inulin during direct fermentation. The sugar purity of DFA III was increased up to 95% by additional incubation with Kluyveromyces marxianus. This study is the first report of the secretory production of recombinant IFTase in Saccharomyces cerevisiae , and the direct conversion of DFA III from crude extract of Jerusalem artichoke tuber powder by microbial fermentation. [ABSTRACT FROM AUTHOR]

Difructose anhydride III (DFAIII) is a prebiotic involved in the reduction of secondary bile acids (BAs). We investigated whether DFAIII modulates BA metabolism, including enterohepatic circulation, in the rats fed with a diet supplemented with cholic acid (CA), one of the 12α-hydroxylated BAs. After acclimation, the rats were fed with a control diet or a diet supplemented with DFAIII. After 2 weeks, each group was further divided into two groups and was fed diet with or without CA supplementation at 0.5 g/kg diet. BA levels were analyzed in aortic and portal plasma, liver, intestinal content, and feces. As a result, DFAIII ingestion reduced the fecal deoxycholic acid level via the partial suppression of deconjugation and 7α-dehydroxylation of BAs following CA supplementation. These results suggest that DFAIII suppresses production of deoxycholic acid in conditions of high concentrations of 12α-hydroxylated BAs in enterohepatic circulation, such as obesity or excess energy intake. Abbreviation: BA: bile acid; BSH: bile salt hydrolase; CA: cholic acid; DCA: deoxycholic acid; DFAIII: difructose anhydride III; MCA: muricholic acid; MS: mass spectrometry; NCDs: non-communicable diseases; LC: liquid chromatography; SCFA: short-chain fatty acid; TCA: taurocholic acid; TCDCA: taurochenodeoxycholic acid; TDCA: taurodeoxycholic acid; TUDCA: tauroursodeoxychlic acid; TαMCA: tauro-α-muricholic acid; TβMCA: tauro-β-muricholic acid; TωMCA: tauro-ω-muricholic acid Suppressed deconjugation and 7α-dehydroxylation of 12α-hydroxylated bile acids by difructose anhydride III contribute to reduction of deoxycholic acid. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • A recombinant IFTase was identified from Arthrobacter chlorophenolicus A6. • The highest activity (902 U mg−1) was shown at 65 °C and pH 5.5. • It showed remarkable thermostability and was stable at up to 80 °C for 1 h. • The conversion ratio of inulin to DFA III was 81% when 50 g l−1 inulin was used. Abstract The gene encoding difructose anhydride III (DFA III)-forming inulin fructotransferase (IFTase) from Arthrobacter chlorophenolicus A6 was cloned and overexpressed in Escherichia.coli. The recombinant IFTase (DFA III-forming) was purified using one-step nickel affinity chromatography. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration analyses, the enzyme showed a homotrimeric form composed of three identical subunits, each with an apparent molecular mass of 43 kDa. The maximum catalytic activity was shown at 65 °C and pH 5.5, and the specific activity was measured as 902 U mg−1. The enzyme showed remarkable thermostability and over half of the initial activity was retained after incubation at 80 °C for 1 h. The K m and V max were estimated to 12.93 mM and 2.89 μmol min−1 ml−1, respectively. After complete hydrolysis of inulin for DFA III production by the purified enzyme, the produced minor products contained sucrose (GF), 1-kestose (GF 2), and nystose (GF 3). The smallest substrate was identified to be GF 3. When 50, 100, and 200 g l−1 of inulin were hydrolyzed by the purified IFTase, the DFA III yield reached 81%, 72%, and 67%, respectively. [ABSTRACT FROM AUTHOR]

Requirements to control the large decrease in serum calcium (Ca) due to parturition and to increase the feed intake soon after parturition have been well accepted in dairy cows. This study was aimed to investigate the feed intake affected by serum Ca concentration with difructose anhydride (DFA) III supplement in dairy cows soon after parturition. Fourteen transition Holstein cows were divided into DFA and control (CONT) groups within 1 to 5 parity variations in each group. Measurement schedule for an individual cow was from 14 d before parturition to 7 d following parturition. The cows in DFA group were supplied 0.2 kg/head/d of DFA III feed containing 40 g of pure DFA III while the cows in CONT group received no DFA III. Other feeding procedures were the same for all cows in both groups. At parturition (d 0), serum Ca concentration sharply declined in both groups (p9.0 mg/dL) tended to be faster in DFA group (12 h) than in the CONT group (48 h), but the differences were not significant. Active ruminal contraction was observed in DFA group at following parturition of d 1 (p

The aim of the present study was to examine the effect of 28 days of dietary difructose anhydride (DFA) III supplementation on calcium (Ca) metabolism in late-lactation dairy cows. Twenty-four multiparous pregnant Holstein cows were divided into two groups. The DFA group was fed total mixed ration (TMR) supplemented with 40 g of DFA III, and the control group was fed TMR only. The replenishment of bone Ca reserves was evaluated by measuring bone mineral density (BMD) and blood biochemical bone markers. Serum Ca concentrations, urinary Ca-to-creatinine (Cre) (Ca/Cre) ratios, and milk Ca concentrations were also analyzed. The BMD of the 4th caudal vertebra in the DFA group was higher than in the control group on day 28. With respect to bone markers, the ratios of undercarboxylated osteocalcin (ucOC) to osteocalcin (OC) in the DFA group were significantly lower than those in the control group on days 21 and 28. Milk Ca concentrations in the DFA group were also higher than those in the control group on days 14, 21, and 28, whereas serum Ca concentrations and urinary Ca/Cre ratios were unchanged in both groups. These results suggest that dietary supplementation with DFA III increased BMD and decreased serum ucOC/OC ratios in late-lactation dairy cows; this indicates that the replenishment of bone Ca reserves may be enhanced by dietary DFA III supplementation. [ABSTRACT FROM AUTHOR]

This experiment was performed to investigate the effects of increases in passively acquired immunoglobulin G (IgG) by difructose anhydride (DFA) III supplementation on subsequent serum IgG concentration and health status in calves during the preweaning period. Thirty newborn female Holstein calves were paired by birth order, and 2 calves in each pair were fed 2 L of the same batch of colostrum within 2 h and at 10 h after birth, and followed by 2 L of the same batch of pooled colostrum at 20 h after birth. One calf from each pair was assigned to the control (n = 15) or treatment (n = 15) group. All calves in the treatment group received 18 g of DFA III at each feeding from birth to 7 d of age, whereas calves in the control group did not receive DFA III. Blood samples were collected before feeding at 0, 10, 20, and 36 h, and 4 and 7 d of age, and sampling was repeated at 7-d intervals thereafter until 49 d of age for serum IgG analysis. Calves were monitored daily for diarrhea and respiratory diseases. Serum IgG concentrations peaked at 36 h of age in both groups. Apparent efficiency of IgG absorption and peak serum IgG concentration were higher in the treatment group than in the control group. Using multiple regression analysis, we showed that peak serum IgG concentration in the newborn calves was positively correlated with colostral IgG concentration and DFA III supplementation. Moreover, peak serum IgG concentration (36 h of age) positively influenced subsequent serum IgG concentration until 35 d of age for all calves in both groups. The treatment group had higher serum IgG concentration from 20 h to 21 d of age than the control group. However, we detected no differences between the groups in number of calves with diarrhea or respiratory disease. [ABSTRACT FROM AUTHOR]

We investigated the effects of dietary supplementation of difructose anhydride III (DFA III), raffinose (Raf), and fructooligosaccharides (FOS) on diet-induced obesity development. Male rats were fed normal or high-fat and high-sucrose (HFS) diet, with or without supplementing (3%) DFA III, Raf, or FOS, for 8 or 5 weeks. Supplementing DFA III to the HFS diet decreased energy intake compared to the non-supplemented HFS diet. Accordingly, body weight gain and fat accumulation reduced in DFA III-fed rats. Cecal acetate production and plasma glucagon-like peptide-1 (GLP-1) and peptide-YY (PYY) were elevated in DFA III-fed rats, while Raf and FOS partially affected these parameters. These results demonstrate that DFA III has suppressive effect on excessive energy intake driven by the palatable obesogenic diet, possibly due to combined effects of increased anorexigenic factors such as cecal acetate production and GLP-1/PYY secretion. [ABSTRACT FROM AUTHOR]

We evaluated the effects of difructose anhydride III (DFAIII) on body weights of ovariectomized rats, which are a good model for obesity by estrogen deficiency-induced overeating. Female rats (10 weeks old) were subjected to ovariectomy or sham operation and then fed with or without a diet containing 3% or 6% DFAIII for 33 days or pair-fed control diet during the same period. Rats fed DFAIII showed significantly decreased food intake, energy intake, body weight gain, body energy accumulation, and fat tissue weight than control group, regardless of ovariectomy. DFAIII may decrease body fat dependent of reduced food/energy intake. Compared with the respective pair feeding groups, rats fed DFAIII showed significantly decreased body energy and fat tissue weight, regardless of ovariectomy, suggesting its potential as a low-energy substitute for high-energy sweeteners. The low energy of DFAIII may contribute to decreased body fat, which may not be dependent on obesity. [ABSTRACT FROM PUBLISHER]

Difructose anhydride (DFA) III acts on intestinal epithelial tight junctions and promotes calcium absorption through the paracellular pathway in dairy cows. This trial was done to investigate the effects of DFA III supplementation on passive IgG transfer and serum IgG concentration in calves. Twenty-four newborn Holstein male calves were grouped as 12 pairs, with each pair receiving a separate batch of pooled colostrum (PC) from the first 2 milkings. One calf from each pair was allocated to the treatment group (n = 12) and received 18 g of DFA III in each feeding; the other calf was assigned to the control group (n = 12) and did not receive DFA III. All calves were fed 2 L of PC per feeding at 1, 10, and 24 h after birth. Prefeeding blood samples taken at 0 h, 10 h, 24 h, 36 h, 4 d, and 7 d were analyzed for serum IgG concentration. Apparent efficiency of IgG absorption in the treatment group was higher compared to that in the control group. Serum IgG concentration of calves in the treatment group was higher for all sampling time points except for 0 h of age. Mean serum IgG concentrations in the treatment and control groups at 36 h of age were 22.3 and 17.7 g/L, respectively. Serum IgG concentration at 36 h of age was positively related with the IgG concentration in the PC in both groups, and it was higher in the treatment group than in the control group at the same IgG concentration in the PC. These results indicate that absorption of ingested IgG by endocytosis as well as a nonselected concentration gradient process of the paracellular pathway of the intestinal epithelium could be enhanced by DFA III supplementation. Supplementation of DFA III in PC containing various IgG concentrations could improve IgG absorption as well as calf serum IgG concentration. [ABSTRACT FROM AUTHOR]

Requirements to control the large decrease in serum calcium (Ca) due to parturition and to increase the feed intake soon after parturition have been well accepted in dairy cows. This study was aimed to investigate the feed intake affected by serum Ca concentration with difructose anhydride (DFA) III supplement in dairy cows soon after parturition. Fourteen transition Holstein cows were divided into DFA and control (CONT) groups within 1 to 5 parity variations in each group. Measurement schedule for an individual cow was from 14 d before parturition to 7 d following parturition. The cows in DFA group were supplied 0.2 kg/head/d of DFA III feed containing 40 g of pure DFA III while the cows in CONT group received no DFA III. Other feeding procedures were the same for all cows in both groups. At parturition (d 0), serum Ca concentration sharply declined in both groups (p<0.05). Time interval for recovery from decreased serum Ca to its normal range (>9.0 mg/dL) tended to be faster in DFA group (12 h) than in the CONT group (48 h), but the differences were not significant. Active ruminal contraction was observed in DFA group at following parturition of d 1 (p<0.05), d 3 (p<0.05), and d 5 (p<0.01). Dry matter (DM) intake did not differ between the groups. However, positive correlations were observed between serum Ca concentration and ruminal contraction (p<0.001), and between ruminal contraction and DM intake (p<0.001) during following parturition. According to multiple regression analysis (R2 = 0.824, p<0.001), the DM intake was positively affected by serum Ca concentration and ruminal contraction. These results suggest that feed intake soon after parturition in dairy cows can be increased by improvement of serum Ca concentration and active ruminal contraction, but DFA III supplementation in this study did not improve the lower serum Ca concentration due to parturition. [ABSTRACT FROM AUTHOR]

Inulin fructotransferase (IFTase; EC 4.2.2.18) has received great attention mainly due to its application in producing difructose anhydride III (DFA III), which is a novel functional sweetener. The object of this study was to investigate the induction of IFTase in Arthrobacter aurescens SK 8.001 with various carbon sources, especially inulin and DFA III. IFTase production could be significantly promoted by the supplement of inulin (5-50 g/L) and DFA III (5-20 g/L). Inulin at high initial concentrations gave no indication of catabolite repression, whereas 30 and 40 g/L DFA III intensely inhibited cell growth and IFTase activity. No fructose was detected in broth throughout the cultivation with inulin, and inulin was converted into DFA III and minor fructooligosaccharides. And when DFA III was the carbon source, DFA III was the only sugar detected in the broth. In conclusion, both DFA III and inulin are effective for IFTase induction, and inulin with higher IFTase activity proved to be a more potent inducer. [ABSTRACT FROM AUTHOR]

Abstract: Objectives: We examined the intestinal segment responsible for and the mechanisms involved in the non-digestible disaccharide difructose anhydride III (DFAIII)-mediated enhancement of iron absorption. Methods: Net iron and calcium absorption (percentage) were observed in male Sprague-Dawley rats fed an AIN93G-based diet with or without DFAIII (30 g/kg of diet) for 2 wk after cecectomy or laparotomy (experiment 1). The absorption rates of calcium and iron from ligated jejunal loops (experiment 2) or ligated cecal sacs (experiment 3) in anesthetized rats fed a control or DFA diet were observed with or without the addition of DFAIII to the instilled mucosal fluid. Levels of the major iron transporter divalent metal transporter-1 (DMT-1) mRNA per 28S rRNA were evaluated by real-time polymerase chain reaction in the intestinal mucosa. Results: Net absorption (percentage) of iron and calcium was higher in the DFA group than in the DFA-free group in laparotomized rats. The enhancement of iron absorption was almost and calcium absorption was partly abolished by cecectomy. The absorption rate of calcium, but not of iron, from the jejunal loops was increased by the addition of DFAIII (100 mmol/L). Iron absorption was clearly higher in the ligated cecal sacs of the DFAIII-fed group than in the DFA-free group with or without the mucosal addition of DFAIII. DMT-1 mRNA level per 28S rRNA was maintained in the cecal mucosa through large increases in cecal size due to DFAIII feeding. Conclusion: The DFAIII-induced increases in iron absorption are the result of increased cecal iron absorptive capacity through expansion of the cecal mucosa maintaining DMT-1 mRNA expression. [Copyright &y& Elsevier]

The article investigates the effects of a nondigestible disaccharide difructose anhydride III (DFAIII) on calcium absorption and retention using a human balance study of single-blind crossover design. According to the authors, the apparent calcium absorption and rate of absorption were higher, and those of retention were much higher, in the DFAIII group than in the control group. They add DFAIII ingestion improves intestinal calcium absorption, which might be beneficial for bone metabolism.

Objective. Total gastrectomy produces osteopenia with calcium malabsorption. We previously demonstrated that difructose anhydride III (DFAIII), a non-digestible disaccharide, stimulates intestinal calcium absorption in normal and ovariectomized rats. In the present study, we examined the effects of feeding DFAIII on gastrectomy-induced calcium malabsorption and osteopenia in rats. The potential of DFAIII to promote large intestinal calcium absorption was also evaluated through comparison with that of fructooligosaccharides (FOS). Material and methods. Male Sprague-Dawley rats were divided into two groups: totally gastrectomized and sham-operated rats. After a postoperative recovery period, rats from each group were divided into three subgroups and fed the control, DFAIII (30 g/kg), or FOS (30 g/kg) diet for 28 days. Results. Total gastrectomy severely reduced net calcium absorption, femoral calcium content and bone mineral density, resulting in fragility of the femur. DFAIII or FOS feeding partly and similarly restored the lowered calcium absorption and femoral variables, with an increase in the total short-chain fatty acid pool in the cecum. In gastrectomized rats, net calcium absorption was correlated with several cecal parameters, suggesting that cecal fermentation of DFAIII is associated with the improvement in gastrectomy-induced calcium malabsorption. Urinary excretion of deoxypyridinoline (D-Pyr) as a marker of bone resorption was increased by gastrectomy, and the elevated D-Pyr excretion was suppressed by feeding DFAIII. Conclusions. Supplemental feeding of DFAIII partly prevents postgastrectomy osteopenia as a result of an improvement in calcium absorption. Our results suggest that the promotive effects of DFAIII on calcium absorption in the large intestine are comparable to those of FOS. [ABSTRACT FROM AUTHOR]

The effects of different kinds of inulin-type fructans on caecal microbiota were evaluated in rats. Four groups of male Wistar rats were fed either a control diet, or diets containing 5% inulin, 5% fructooligosaccharides (FOS), or 5% difructose anhydride III (DFAIII) for two weeks. In the DFAIII group, caecal propionate, butyrate, counts of bifidobacteria, and total anaerobes were lower than in the inulin group, while caecal propionate, succinate, counts of bifidobacteria, and total anaerobes were lower than in the FOS group. Compared to controls, in the DFAIII group the counts of clostridia in caecum were increased by 3 log units. However, this change was statistically not significant. There were no differences between inulin and FOS groups for the pool of short chain fatty acids in caecum and bacterial counts. Results indicate that DFAIII has different effects on caecal microbiota compared to inulin and FOS and that these differences are most likely due to the α(3→2) bonds in DFAIII. [ABSTRACT FROM AUTHOR]

Abstract: Objective: Total gastrectomy produces iron malabsorption and anemia, and several non-digestible carbohydrates promote mineral absorption. In this study, we examined the effects of feeding difructose anhydride III (DFAIII), a non-digestible disaccharide, on gastrectomy-induced iron malabsorption and anemia in rats in comparison with those of feeding fructo-oligosaccharides (FOS). Methods: Sham-operated and totally gastrectomized male Sprague-Dawley rats were fed the control, DFAIII (30 g/kg), or FOS (30 g/kg) diet for 4 wk. Feces and tail blood were collected at 2 and 4 wk to evaluate body iron status and iron absorption. Results: Gastrectomy severely decreased net iron absorption, hemoglobin concentration, and hematocrit in the control dietary group. The decreased absorption in gastrectomized rats was restored to the sham control level by feeding the DFAIII or FOS diet. Iron absorption in sham rats was higher in the FOS and DFAIII groups than in the control group. Hemoglobin concentration and hematocrit in gastrectomized rats fed the DFAIII diet, but not the FOS diet, returned to levels comparable to the effects in sham rats fed the control diet. Feeding DFAIII increased short-chain fatty acid pools and decreased pH of cecal contents. These parameters for cecal fermentation correlated with iron absorption. Conclusions: DFAIII feeding restores gastrectomy-induced iron malabsorption, resulting in complete prevention of iron-deficiency anemia in rats. Cecal fermentation of DFAIII may contribute to the improvement in these gastrectomy-induced defects. Feeding with low level of FOS did not fully improve postgastrectomy anemia. [Copyright &y& Elsevier]

Abstract: A nondigestible disaccharide, difructose anhydride (DFA) III, is known to activate calcium transport via tight junctions (TJs); however, the characteristics of and mechanisms for the increase in paracellular transport induced by DFAIII have not been clarified. We compared the effect of DFAIII with that of sodium caprate (C10), a well-known enhancer of TJ permeability, on the changes in TJ proteins, transport of paracellular markers, and effects of nine cellular signaling blockers using Caco-2 monolayers. The addition of DFAIII (0–100mmol/L) and C10 (0–10mmol/L) to the apical medium of the Caco-2 monolayers dose-dependently decreased transepithelial electrical resistance (TER), which is an indicator of TJ permeability. The reduction with C10 was much faster than that with DFAIII. Transport of the paracellular markers of various molecular weights (182–43,200) was elevated by the addition of 100mmol/L DFAIII and 10mmol/L C10. The transport rates were much in the presence of C10 than of DFAIII, while the reduction in TER by two treatments was similar (from 1000 to 300Ω cm2). Treatment with DFAIII and C10 changed the distribution of actin filament and claudin-1, but not occludin, junctional adhesion molecule-1, or zonula occludens-1; however, alterations in the patterns of the TJ proteins differed according to treatment. An inhibitor of myosin light chain kinase and a chelator of intracellular calcium ion ([Ca2+]i) attenuated the TER reduction by C10, but not by DFAIII. These data demonstrate that the increase in TJ permeability induced by DFAIII results from the alterations to actin and claudin-1 via [Ca2+]i-independent mechanisms. [Copyright &y& Elsevier]

The article presents a study which evaluated the body energy accumulation as fat and protein from ingestion of difructose anhydride III (DFAIII). Supplements of DFAIII did not increase whole body or peripheral fat or total body energy while sucrose increased them in a dose-dependent manner. The results show that DFAIII is a fermentable saccharide with low available energy for fat accumulation.

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. [ABSTRACT FROM PUBLISHER]

This study was conducted to investigate the effect of difructose anhydride (DFA) III on dry matter intake in multiparous cows immediately after calving. Seventy-four transition multiparous Holstein cows were allocated into control (CONT) and DFA groups. The average parity number in the CONT and DFA groups was 3.70 ± 0.25 and 3.67 ± 0.26, respectively. Individual cow measurements were collected from 14 d before calving to 7 d after calving. All cows were fed a total mixed ration and grass hay before calving, and were fed another total mixed ration and grass hay after calving. The cows in the CONT group received no DFA III during the experimental period, and the cows in the DFA group received 40 g/d of DFA III from 14 d before to 7 d after calving. Dry matter intake was measured throughout the experiment. Serum calcium concentration, ruminal contraction rate and body weight were measured before and after calving. Milk yield was recorded after calving. The dry matter intake did not differ between the two groups before calving, and the lowest dry matter intake was observed at calving in the both groups. The dry matter intake began to increase after calving in the both groups but was higher in the DFA group (P < 0.01). At calving, the serum calcium concentration decreased to approximately 7.4 mg/dl in the both groups. However, the cows in the DFA group showed faster recovery in the serum calcium concentration from the decline to above 9.0 mg/dl. The ruminal contraction rate did not differ between the two groups and decreased at calving in the both groups. After calving, the ruminal contraction rate began to recover in the both groups but was greater in the DFA group. The ruminal contraction rate after calving was positively associated with the serum calcium concentration after calving (P < 0.01). Average milk yield after calving was higher in the DFA group than in the CONT group (P < 0.05). The average dry matter intake for the first 7 d after calving was positively associated with the serum calcium concentration, frequency of ruminal contraction and milk yield after calving (P < 0.01). These results showed that DFA III enhances the ruminal contraction rate and increases dry matter intake immediately after calving in multiparous cows by quickly restoring the serum calcium concentrations after their decline at calving. [ABSTRACT FROM AUTHOR]

The aim of the present study was to examine the effect of 28 days of dietary difructose anhydride (DFA) III supplementation on calcium (Ca) metabolism in late-lactation dairy cows. Twenty-four multiparous pregnant Holstein cows were divided into two groups. The DFA group was fed total mixed ration (TMR) supplemented with 40 g of DFA III, and the control group was fed TMR only. The replenishment of bone Ca reserves was evaluated by measuring bone mineral density (BMD) and blood biochemical bone markers. Serum Ca concentrations, urinary Ca-to-creatinine (Cre) (Ca/Cre) ratios, and milk Ca concentrations were also analyzed. The BMD of the 4th caudal vertebra in the DFA group was higher than in the control group on day 28. With respect to bone markers, the ratios of undercarboxylated osteocalcin (ucOC) to osteocalcin (OC) in the DFA group were significantly lower than those in the control group on days 21 and 28. Milk Ca concentrations in the DFA group were also higher than those in the control group on days 14, 21, and 28, whereas serum Ca concentrations and urinary Ca/Cre ratios were unchanged in both groups. These results suggest that dietary supplementation with DFA III increased BMD and decreased serum ucOC/OC ratios in late-lactation dairy cows; this indicates that the replenishment of bone Ca reserves may be enhanced by dietary DFA III supplementation.