Prebiotic effect of fructooligosaccharide in the simulator of the human intestinal microbial ecosystem (SHIME® model).

Maintaining "gut health" is a goal for scientists throughout the world. Therefore, microbiota management models for testing probiotics, prebiotics, and synbiotics have been developed. The SHIME(®) model was used to study the effect of fructooligosaccharide (FOS) on the fermentation pattern of the colon microbiota. Initially, an inoculum prepared from human feces was introduced into the reactor vessels and stabilized over 2 weeks using a culture medium. This stabilization period was followed by a 2-week control period during which the microbiota was monitored. The microbiota was then subjected to a 4-week treatment period by adding 5 g/day-1 FOS to vessel one (the "stomach" compartment). Plate counts, Denaturing Gradient Gel Electrophoresis (DGGE), short-chain fatty acid (SCFA), and ammonium analyses were used to observe the influence of FOS treatment in simulated colon compartments. A significant increase (P<.01) in the Lactobacillus spp. and Bifidobacterium spp. populations was observed during the treatment period. The DGGE obtained showed the overall microbial community was changed in the ascending colon compartment of the SHIME reactor. FOS induced increase of the SCFA concentration (P<.05) during the treatment period, mainly due to significant increased levels of acetic and butyric acids. However, ammonium concentrations increased during the same period (P<.01). This study indicates the usefulness of in vitro methods that simulate the colon region as part of research towards the improvement of human health.

Incorporation of soybean by-product okara and inulin in a probiotic soy yoghurt: texture profile and sensory acceptance.

BACKGROUND: This study evaluated the effect of inulin and okara flour on textural and sensory properties of probiotic soy yoghurt (SY) throughout 28 days of storage at 4 °C. Employing a 2(2) design, four formulations of SY produced from soymilk and fermented with an ABT-4 culture (Lactobacillus acidophilus La-5, Bifidobacterium animalis Bb-12 and Streptococcus thermophilus) were studied: SY-C (control); SY-I (with inulin); SY-O (with okara); SY-IO (with inulin + okara). RESULTS: The addition of okara and the refrigerated storage led to significant differences in the instrumental texture parameters of SY (P < 0.05). Inulin and okara did not affect SY sensory acceptability (P > 0.05), but there was a tendency for higher scores in the presence of inulin. On the other hand, the storage period, particularly at 21 days, was unfavourable regarding the acceptance of the different SY. CONCLUSION: The results showed that the addition of okara flour and the storage were significant factors to increase firmness of the soy yoghurts. SY acceptability was not affected by the incorporation of inulin or okara. These results suggest that okara, discarded as industrial waste, may be used in probiotic soy yoghurt, helping to increase the nutritional and functional properties without altering its acceptability.

Lactobacillus acidophilus CRL 1014 improved "gut health" in the SHIME reactor.

BACKGROUND: How to maintain "gut health" is a goal for scientists throughout the world. Therefore, microbiota management models for testing probiotics, prebiotics, and synbiotics have been developed. METHODS: The SHIME model was used to study the effect of Lactobacillus acidophilus 1014 on the fermentation pattern of the colon microbiota. Initially, an inoculum prepared from human feces was introduced into the reactor vessels and stabilized over 2-wk using a culture medium. This stabilization period was followed by a 2-wk control period during which the microbiota was monitored. The microbiota was then subjected to a 4-wk treatment period by adding 5 mL of sterile peptone water with L. acidophilus CRL1014 at the concentration of 108 CFU/mL to vessel one (the stomach compartment). Plate counts, Denaturing Gradient Gel Electrophoresis (DGGE), short-chain fatty acid (SCFA) and ammonium analyses were carried out for monitoring of the microbial community from the colon compartments. RESULTS: A significant increase (p < 0.01) in the Lactobacillus spp. and Bifidobacterium spp. populations was observed during the treatment period. The DGGE obtained showed changes in the lactobacilli community from the colon compartments of the SHIME reactor. The (SCFA) concentration increased (p < 0.01) during the treatment period, due mainly to significant increased levels of acetic, butyric, and propionic acids. However, ammonium concentrations decreased during the same period (p < 0.01). CONCLUSIONS: This study showed the beneficial influence of L. acidophilus CRL 1014 on microbial metabolism and lactobacilli community composition for improving human health.

A potential synbiotic product improves the lipid profile of diabetic rats.

BACKGROUND: Previous studies showed that intake of yacon or some lactic acid bacteria was able to inhibit the development of diabetes mellitus, by reducing glucose and associated symptoms, for example, the lipid profile. OBJECTIVE: The purpose of this study was to assess the consumption influence of a potential symbiotic product of soybean and yacon extract and fermented Enterococcus faecium CRL 183 and Lactobacillus helveticus ssp jugurti 416 in reducing blood glucose and lipid levels in an animal model. METHODS: Diabetes mellitus was chemically induced by intraperitoneal administration of streptozotocin (50 mg/kg body weight). The rats were divided into four groups (n=10): GI - non-diabetic animals that received only a standard chow diet (negative control), GII - diabetic animals that received only chow diet (positive control), GIII - diabetic animals that received the chow diet + 1 mL/kg body weight/day of soybean and yacon unfermented product, GIV - diabetic rats that received the chow diet + 1 mL/kg body weight/day of soybean and yacon fermented product. There was a seven-week treatment period and the following parameters were evaluated: animal body weight, food and water intake, blood glucose, enzyme activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), triglycerides levels, total cholesterol, HDL-C, non-HDL-C. Cell viability of the fermented product was checked weekly for a seven-week period. RESULTS: The product average viable population was 10(8)-10(9) CFU/mL, by ensuring both the rods and cocci regular intake. No difference was observed between the water and feed intake and body weight of groups that received unfermented and fermented products and the untreated diabetic group. The same was observed for the blood glucose and AST and ALT activities, while some improvement was observed for a lipid profile, represented by reduction of triglycerides level by 15.07% and 33.50% in groups III and IV, respectively, and an increase of 23.70% in HDL-C level for group IV. CONCLUSION: The results showed that the ingestion of a potential symbiotic product was neither able to promote improvement in some of the disease symptoms, nor reduce blood glucose. However, a positive effect on triglycerides levels and HDL-cholesterol was observed in the groups that received the unfermented product containing yacon extract and the fermented product with Enterococcus faecium CRL 183, as well as Lactobacillus helveticus ssp jugurti 416 and yacon extract (symbiotic product).

Influence of a probiotic soy product on fecal microbiota and its association with cardiovascular risk factors in an animal model.

BACKGROUND: Previous work showed that daily ingestion of an aqueous soy extract fermented with Enterococcus faecium CRL 183 and Lactobacillus helveticus 416, supplemented or not with isoflavones, reduced the total cholesterol and non-HDL-cholesterol levels, increased the high-density lipoprotein (HDL) concentration and inhibited the raising of autoantibody against oxidized low-density lipoprotein (ox-LDL Ab) and the development of atherosclerotic lesions. OBJECTIVE: The aim of this study was to characterize the fecal microbiota in order to investigate the possible correlation between fecal microbiota, serum lipid parameters and atherosclerotic lesion development in rabbits with induced hypercholesterolemia, that ingested the aqueous soy extract fermented with Enterococcus faecium CRL 183 and Lactobacillus helveticus 416. METHODS: The rabbits were randomly allocated to five experimental groups (n = 6): control (C), hypercholesterolemic (H), hypercholesterolemic plus unfermented soy product (HUF), hypercholesterolemic plus fermented soy product (HF) and hypercholesterolemic plus isoflavone-supplemented fermented soy product (HIF). Lipid parameters and microbiota composition were analyzed on days 0 and 60 of the treatment and the atherosclerotic lesions were quantified at the end of the experiment. The fecal microbiota was characterized by enumerating the Lactobacillus spp., Bifidobacterium spp., Enterococcus spp., Enterobacteria and Clostridium spp. populations. RESULTS: After 60 days of the experiment, intake of the probiotic soy product was correlated with significant increases (P < 0.05) on Lactobacillus spp., Bifidobacterium spp. and Enterococcus spp. and a decrease in the Enterobacteria population. A strong correlation was observed between microbiota composition and lipid profile. Populations of Enterococcus spp., Lactobacillus spp. and Bifidobacterium spp. were negatively correlated with total cholesterol, non-HDL-cholesterol, autoantibodies against oxidized LDL (oxLDL Ab) and lesion size. HDL-C levels were positively correlated with Lactobacillus spp., Bifidobacterium spp., and Enterococcus spp. populations. CONCLUSION: In conclusion, daily ingestion of the probiotic soy product, supplemented or not with isoflavones, may contribute to a beneficial balance of the fecal microbiota and this modulation is associated with an improved cholesterol profile and inhibition of atherosclerotic lesion development.

Effect of fermented soy product on the fecal microbiota of rats fed on a beef-based animal diet.

BACKGROUND: The aim of this study was to determine whether the consumption of soy product fermented with Enterococcus faecium CRL 183, would modify the fecal microbiota of rats fed a diet containing red meat. The rats were placed in groups, distinguished by their diets. For 60 days, group I was given a standard casein-based rodent feed and groups II-VI, the beef-based feed. From the 30th day, groups III-VI also ingested the following products: group III, E. faecium-fermented soy product; group IV, pure suspension of E. faecium; group V, sterilized fermented soy product; and group VI, unfermented soy product. RESULTS: Rats that ingested fermented soy product showed a slight increase in the numbers of lactobacilli (0.45 log CFU g(-1)), as did the casein-based diet group (0.47 log CFU g(-1)). The fermented soy product did not cause any reduction in the number of enterobacteria or clostridia, but promoted a slight fall in the viable count of Bacteroides spp. (2.80 +/- 0.20 to 2.34 +/- 0.07 log CFU g(-1)). CONCLUSIONS: The results indicate that the ingestion of this fermented soy product did not lead to significant changes in the fecal microbiota of the rats fed on a beef-based diet.

Effects of isoflavone-supplemented soy yogurt on lipid parameters and atherosclerosis development in hypercholesterolemic rabbits: a randomized double-blind study.

BACKGROUND: There is increasing interest in natural treatments to control dyslipidemia and reduce the risk of cardiovascular disease. Previous studies have demonstrated the beneficial effects of soy yogurt fermented with Enterococcus faecium CRL 183 and of dietary isoflavones on the lipid profile. The purpose of the present study was to investigate the effects of isoflavone-supplemented soy yogurt, fermented with E. faecium CRL183, on lipid parameters and atherosclerosis development in rabbits with induced hypercholesterolemia. METHODS: Forty-eight rabbits were randomly assigned to eight groups fed on the following diets for 60 days: C - control; IY - isoflavone-supplemented soy yogurt; H - hypercholesterolemic (1.0% cholesterol wt/wt diet); HY - hypercholesterolemic plus soy yogurt; HIY - hypercholesterolemic plus isoflavone-supplemented soy yogurt; HP - hypercholesterolemic plus placebo; HI - hypercholesterolemic plus isoflavone and HE - hypercholesterolemic plus pure culture of E. faecium CRL 183. Serum lipids and autoantibodies against oxLDL (oxLDL Ab) were analyzed on days 0, 30 and 60 of the treatment and the atherosclerotic lesions were quantified at the end of the experiment. RESULTS: Soy yogurt, soy yogurt supplemented with isoflavones and placebo promoted significant reductions in total cholesterol level (38.1%, 27.0% and 26.6%, respectively). Significant increases in serum HDL-C concentration relative to group H were detected in animals that ingested soy yogurt, with or without the isoflavone supplement (55.2%), E. faecium culture (43.3%) or placebo (35.8%). Intake of soy yogurt and soy yogurt supplemented with isoflavones prevented the rise of oxLDL Ab during the study period. The extent of atherosclerosis in the thoracic and abdominal aortas was reduced in the HIY, HY and HP groups. However, when the whole aorta was analyzed, animals treated with soy yogurt supplemented with isoflavones exhibited the greatest reduction (51.4%, P < 0.05) in atherosclerotic lesion area, compared to group H. CONCLUSION: Soy yogurt could be consumed as an alternative means of reducing the risk of cardiovascular disease by improving the lipid profile and inhibiting oxLDL Ab formation. Our findings also suggest that isoflavone supplementation may enhance the antiatherosclerotic effect of soy yogurt.

Effects of a probiotic soy product and physical exercise on formation of pre-neoplastic lesions in rat colons in a short-term model of carcinogenic.

PURPOSE: In this study the influence of moderate or intense physical exercise, alone or in combination with the consumption of a soya product fermented with Enterococcus faecium, on the development of colon cancer induced chemically in rats with 1,2-dimethylhydrazine (DMH), was investigated. METHODS: Eighty male Wistar SPF rats were randomly allocated to 8 groups (n = 10). One week after the start of the program of product ingestion and/or physical activity, all animals except the controls (group I) were injected subcutaneously with 50 mg/kg b.w. of 1,2-dimethylhydrazine (DMH). This procedure was repeated at the end of the second week. At the end of the 6-week experiment, all the animals were euthanized; the colons were removed and numbers of ACF was estimated. RESULTS: Twenty-four days after the induction of pre-neoplastic lesions, it was evident that the formation of ACF was not significantly reduced by the ingestion of the fermented product, by intense or moderate physical activity or by a combination of these factors, in comparison with the positive control group of rats (p < 0.05). On the other hand, the performance of intense exercise, on its own, increased the number of ACF. CONCLUSION: The results reported in this article show that consumption of the fermented soy product described here and the practice of physical exercise (intense or moderate) were incapable, separately or combined, of inhibiting the formation of ACF in DMH-induced rats. The intense physical exercise led to an increased number of foci in the colons of these rats and, probably, to greater susceptibility to colorectal cancer.

Effects of probiotic bacteria, isoflavones and simvastatin on lipid profile and atherosclerosis in cholesterol-fed rabbits: a randomized double-blind study.

BACKGROUND: Much attention has been drawn to different alternative strategies for cardiovascular disease prevention. OBJECTIVE: The aim of the present study was to observe and compare the effects of Enterococcus faecium CRL183 (probiotic microorganism), an isoflavones mixture and simvastatin (drug used to treat hypercholesterolemia) on lipid parameters and atherosclerosis development in rabbits with induced hypercholesterolemia. METHODS: The animals were randomly allocated to 5 experimental groups (n = 6) for 60 days: control (C) that did not consume cholesterol, hypercholesterolemic (H) that consumed an atherogenic diet (1.0% cholesterol wt/wt), hypercholesterolemic plus E. faecium (HE), hypercholesterolemic plus isoflavone (HI) and hypercholesterolemic plus simvastatin (HS). Total and HDL-cholesterol and triglycerides were determined by enzymatic methods; non-HDL-C was calculated by subtracting HDL-C from total cholesterol; and atherosclerosis was presented as the percentage of lesion area, relative to the total area from the aorta segment analyzed. RESULTS: Simvastatin significantly reduced the tot cholesterol (16%) and non-HDL-C level (17%) and increased the HDL-C (98%), compared to group H. E. faecium raised the HDL-C level by 43.3% (P < 0.05). Isoflavone decreased the total cholesterol and non-HDL-C concentrations (9%), but this effect was not statistically significant. At the end of the treatments, groups HE and HS had significantly lower levels of triglycerides in relation to H and HI. The atherosclerotic lesion area in the aortic arch was not different between groups. The extent of atherosclerosis in the thoracic and abdominal aorta was reduced in the groups HI and HS by 22.7% and 26.7% respectively, but this effect was not significant (P > 0.05). CONCLUSION: The results indicated that probiotic microorganism E. faecium CRL 183 could be used to improve the lipid profile as an alternative or an adjuvant for drug therapy. The effectiveness of simvastatin in the management of blood lipid was confirmed. There were no effects of soy isoflavones, E. faecium and simvastatin on atherosclerosis development.

Effects of 2 or 5 consecutive exercise days on adipocyte area and lipid parameters in Wistar rats.

BACKGROUND: Exercise has been prescribed in the treatment and control of dyslipidemias and cholesterolemia, however, lipid responses to different training frequencies in hypercholesterolemic men have been inconsistent. We sought to verify if different frequencies of continuous moderate exercise (2 or 5 days/week, swimming) can, after 8 weeks, promote adaptations in adipocyte area and lipid parameters, as well as body weight and relative weight of tissues in normo and hypercholesterolemic adult male rats. METHODS: Normal cholesterol chow diet or cholesterol-rich diet (1% cholesterol plus 0.25% cholic acid) were freely given during 8 weeks to the rats divided in 6 experimentals groups: sedentary normal cholesterol chow diet (C); sedentary cholesterol-rich diet (H); 5x per week continuous training normal cholesterol chow diet (TC5) and cholesterol-rich diet (TH5); 2x per week continuos traning normal cholesterol chow diet (TC2) and cholesterol-rich diet (TH2). RESULTS: No changes were observed in lipid profile in normal cholesterol chow diet, but both 2 a 5 days/week exercise improved this profile in cholesterol-rich diet. Body weight gain was lower in exercised rats. Decrease in retroperitoneal and epididymal relative weights as well as reductions in adipocyte areas under all diets types were observed only in 5 days/week, while 2 days/week showed improvements mainly in cholesterol-rich diet rats. CONCLUSION: Our results confirm the importance of exercise protocols to control dyslipidemias and obesity in rats. The effects of 5 days/week exercise were more pronounced compared with those of 2 consecutive days/week training.

Fermented soy product supplemented with isoflavones affected fat depots in juvenile rats.

OBJECTIVES: This study investigated the effects of soy product fermented by Enterococcus faecium and Lactobacillus jugurti supplemented with isoflavones on adipose tissue, blood lipid, and glucose levels on juvenile rats. METHODS: Rats were fed a cholesterol-enriched diet for 3 wk as a preliminary treatment to create hypercholesterolemia. They were then fed a chow diet (HC), a chow diet plus fermented soy product supplemented with isoflavones (HCFI), a chow diet plus placebo (HCP), or a chow diet plus placebo supplemented with isoflavones (HCPI), respectively, for an additional 3 wk. RESULTS: The beneficial effects of fermented soy product supplemented with isoflavones on epididymal (EPI) and retroperitoneal (RET) fat pads was likely due to isoflavones because adipocyte circumference (micrometers) in the HC group was significantly larger (EPI: 105.66 +/- 13.36; RET: 134.95 +/- 25.40) than that in the HCFI group (EPI: 93.17 +/- 12.80; RET: 108.62 +/- 15.50) and HCPI group (EPI: 93.06 +/- 15.10; RET: 112.34 +/- 18.21). The probiotic micro-organism accentuated the antilipogenic effect of isoflavones on RET (HCFI: 108.62 +/- 15.50 micrometers versus HCPI: 112.34 +/- 18.21 micrometers). Moreover, the fermented product increased glucose concentration similar to that in the chow group but did not change blood lipids. CONCLUSION: This product may offer new approaches to obesity prevention.

Efeito de um novo produto fermentado de soja sobre os lípides séricos de coelhos hipercolesterolêmicos / Effects of a novel fermanted soy product on the serum lipids of hypercholesterolemic rabbits

OBJECTIVE: To assess the effect of a new feed soy product fermented by Enterococcus faecium and Lactobacillus jugurti on the serum lipid levels of rabbits with induced hypercholesterolemia. METHODS: Thirty-two rabbits were divided into 4 groups as follows: 1) control (C); 2) hypercholesterolemic (H); 3) hypercholesterolemic + fermented product (HPF); and 4) control + fermented product (CPF). The H and HPF groups were fed with a diet with 0.15 per cent (p/p) cholesterol in the first 15 days. C and CPF groups received regular food preparation. The HPF and CPF groups received 10 mL daily of the fermented 30 days. Blood samples were drawn at the beginning of the study and at the 15th and 30th days. Concentrations of total cholesterol, HDL-cholesterol, and triglycerides were analyzed. RESULTS: After 15 days, the HPF group showed a total cholesterol concentration lower (18.4 per cent) than that of the H group (p=0.05), but this difference disappeared after 30 days. No change was observed in total cholesterol levels of C and CPF groups. After 15 days, the HDL-cholesterol was higher (17.8 per cent) in the HPF group, but the triglyceride levels remained unchanged in all groups during the same period of time. CONCLUSION: The soy fermented product caused an 18.4 per cent reduction in total cholesterol and a 17.8 per cent increase in the HDL-fraction. It may, therefore, be a possible coadjutor in the treatment of hypercholesterolemia.