中性大蒜果聚糖体外发酵产短链脂肪酸

大蒜果聚糖是大蒜深加工后废弃蒜渣中的主要成分,不能被人体消化利用,可选择性增殖肠道益生菌,具有开发成新型益生元的潜力。体外发酵产短链脂肪酸的分析对进一步评价大蒜果聚糖对肠道微生态的影响有重要意义。方法:本研究通过体外模拟人体肠道系统的pH、温度和营养条件,接种人体新鲜粪便悬浮液进行间歇式静态厌氧培养,采用高效液相色谱法(HPLC)分析不同聚合度(平均DP为11、16、21)的中性大蒜果聚糖为唯一碳源,培养0、4、8、12、24 h产短链脂肪酸的情况。结果:发酵12 h时短链脂肪酸浓度达到最高,之后便开始下降。其中样a(平均DP为11)的发酵液中总SCFA浓度最高,达86.38μmol/mL。总短链脂肪酸产量为样a(DP 11)>样b(DP16)>样c(DP 21)。24 h时,各发酵液的短链脂肪酸中丁酸和乳酸的比例较乙酸和丙酸高。结论:大蒜果聚糖能被人体肠道微生物发酵利用在体外产短链脂肪酸。其中丁酸和乳酸的摩尔浓度较乙酸和丙酸高。聚合度低的大蒜果聚糖产酸高于聚合度高的果聚糖。     

植物甾醇酯对高脂饮食大鼠结肠内容物短链脂肪酸的影响

目的:研究植物甾醇酯(phytosterol ester,PSE)对高脂饮食大鼠结肠内容物短链脂肪酸的影响。方法:将30只6周龄雄性SD大鼠随机分为四组,其中正常对照组饲喂普通饲料,高脂组饲喂高脂饲料,PSE干预组分别灌胃低(0.05 g/100 g BW·d)、高剂量PSE (0.10 g/100 g BW·d) PSE强化牛奶;连续干预13周后,处死动物取结肠内容物,利用气相色谱法测定其SCFAs的含量。结果:与正常对照组相比,高脂饮食使大鼠结肠内容物中总SCFAs含量增加33.63%,其中丙酸含量显著升高(126.07%)(p<0.05),乙酸(26.05%)和异戊酸(42.11%)含量具有升高趋势(p>0.05),丁酸(33.21%)和戊酸(23.92%)含量则表现为降低趋势(p>0.05);而PSE干预可抑制高脂饮食所致大鼠结肠内容物中SCFAs水平的升高,其中低剂量PSE可显著降低丙酸(51.52%)、丁酸(59.59%)、异戊酸(60.66%)和戊酸(72.07%)的含量(p<0.05);高剂量PSE干预可降低丁酸(29.48%)的含量(p>0.05),并显著降低乙酸(48.64%)、丙酸(58.39%)、戊酸(69.12%)和异戊酸(58.59%)的含量(p<0.05)。结论:高脂饮食会引起大鼠结肠中SCFAs水平的升高,而PSE可能通过降低结肠内容物SCFAs的含量来调节大鼠肠道内环境。     

Effects of different types of potato resistant starches on intestinal microbiota and short-chain fatty acids under in vitro fermentation

The prebiotic effects of potato resistant starches RS1, RS2, RS3a, RS3b and RS4 were studied using simulated gut fermentation in vitro. The intestinal microbial composition was significantly changed by resistant starch (RS) after 24-h fermentation. All RSs (P < 0.05) decreased the ratio of Firmicutes/Bacteroidetes (F/B), and RS2 exhibited the lowest value of 0.95 ± 0.07. The relative abundance of Bifidobacterium was significantly increased by RS4, whereas Megamonas and Prevotella were promoted by RS2. Further, RS4 produced the highest levels of acetate (138.34 µM), whereas RS2 produced the highest levels of propionate (41.45 µM) and butyrate (21.65 µM). However, Bifidobacterium did not promote the production of propionate or butyrate, even though it was proficient in fermenting RS. Megamonas and Prevotella were positively associated with the higher production of propionate and butyrate. Here, different RSs played key roles in promoting intestinal health, and RS2 especially showed more abundant probiotic functions.     

山药多糖和燕麦多糖发酵产酸及发酵产物对结肠癌细胞的增殖抑制作用

采用水提取、醇沉淀法制备山药多糖和燕麦多糖,利用健康成人粪便提取物中的肠道微生物体外发酵多糖,确定发酵产酸情况（乙酸,丙酸,丁酸,乳酸）以及发酵产物对结肠癌细胞HCT-116的增殖抑制作用。山药多糖和燕麦多糖的糖含量分别为82.63%和80.32%。发酵24 h,山药多糖和燕麦多糖产生23.28、25.14 mmol/L乙酸、2.49、1.97 mmol/L丙酸、11.04、7.99 mmol/L丁酸和0.19、0.46 mmol/L乳酸;发酵48 h,它们产生四种酸为28.14、42.53,6.48、2.45,13.76、9.64,0.08、0.09 mmol/L。发酵时间从24 h延长至48 h,产物中乙酸、丙酸和丁酸含量显著提高（p<0.05）,而乳酸含量明显降低（p<0.05）。两种多糖对细胞的抑制作用小于24.5%,而两种多糖发酵产物有更强的抑制作用,分别达到46.2%⁶9.1%、44.6%⁶7.3%;48 h发酵产物抑制作用更强,处理细胞48 h产生的抑制作用更大。结果表明,两种多糖经过肠道微生物发酵后,其产物对结肠癌细胞具有更强的增殖抑制活性。      

Effects of goat milk enriched with oligosaccharides on microbiota structures,and correlation between microbiota and short-chain fatty acids in the large intestine of the mouse

In this study, we explored the effects of combining goat milk and oligosaccharides on the large intestine environment of mice. A combination of goat milk with each of 3 oligosaccharides—stachyose, fructo-oligosaccharide (FOS), and a prebiotics mix—were independently fed to mice. We investigated composition changes in the microbiota of the large intestine using 16S rRNA gene sequencing; measured short-chain fatty acid content using gas chromatography–mass spectrometry; and performed a Spearman correlation analysis between microorganisms and short-chain fatty acids. Our results showed that microbial diversity in the large intestine decreased significantly in the FOS group. In terms of α diversity, microbial richness significantly declined in all 3 treatment groups; in terms of β diversity, the intestinal microbial structures clearly changed in the FOS group. The abundance of Bifidobacterium and Lactobacillus increased markedly in the FOS group compared with the other groups. Functional predictions showed that FOS reduced intestinal bacterial infections and improved the endocrine and immune systems. Spearman correlation analysis showed that propionic, isobutyric, and valeric acids were all positively correlated with certain microbiota. Our findings suggest that FOS-enriched goat milk is beneficial for improving the large intestine environment in the host.     

Beneficial effects of three brown seaweed polysaccharides on gut microbiota and their structural characteristics: An overview

The gut microbiota appears critical in the metabolic health and anti-disease activity. In this review, we discuss three brown seaweed polysaccharides (alginate, laminarin, fucoidan) for their structural information, the digestive behaviour and the effects on gut microbiota. Bioactivities are associated with various physicochemical properties, like solubility, viscosity, hydration properties, molecular weight, monosaccharide composition, and so on. Brown seaweed polysaccharides can be completely utilised by microbes in the large bowel, and they can regulate the gut microbiota. The ultimate metabolite of these polysaccharides is mainly short-chain fatty acids, which are able to regulate the ecology of the gut microbiota by increasing the growth of beneficial bacteria and inhibiting the growth of some harmful bacteria. In addition, this article also discusses the relationship between the structure and activity of modulating the gut microbiota. Results show that polysaccharides with low molecular weight are more conductive to modulate gut microbiota.     

The effects of polydextrose and xylitol on microbial community and activity in a 4-stage colon simulator

ABSTRACT:  This study focused on the effects of candidate prebiotics polydextrose (PDX) and xylitol on the microbial community and its metabolic activity in a colon simulator. A semicontinuous, anaerobic culture system was used with 4 vessels mimicking the conditions in the human large intestine from proximal to distal colon. Bacterial inocula for the independent simulations were obtained from fecal samples of different donors. Synthetic medium, mimicking the contents of the small intestine, containing either 2% of the prebiotic candidate or no added carbohydrates as a control, was fed to the system. After 48 h of simulation samples were collected and analyzed. A sustained degradation of polydextrose throughout the colon model and a more rapid degradation of xylitol were observed. The fermentation of both compounds was characterized by a significantly increased production of short-chain fatty acids (SCFA). Polydextrose increased the concentrations of all SCFA, especially acetate and propionate, and xylitol especially the concentration of butyrate. Branched-chain fatty acids (BCFA) levels decreased significantly as a result of polydextrose and xylitol supplementation, whereas biogenic amine levels remained mostly unchanged. Thus, a beneficial shift in the metabolic patterns of the colon microbes was measured with both of the tested products. These in vitro studies provide evidence to the prebiotic characteristics of polydextrose; also, further beneficial properties of xylitol were demonstrated in the colon model.     

Effects of direct-fed Bacillus subtilis and Bacillus licheniformis on production performance and milk fatty acid profile in dairy cows

The aim of the study was to determine the effect of a Bacillus-based direct-fed microbial on performance of mid-lactating Holstein dairy cows and on their milk fatty acid composition. Six multiparous cows fitted with a rumen cannula were used in a randomized replicated crossover design. Cows received 200 g/d of either whey powder as a control or BioPlus 2B (Chr. Hansen), a commercial direct-fed microbial providing Bacillus subtilis and Bacillus licheniformis, representing a daily dose of 6.4 × 10¹¹ cfu, and using whey powder as a carrier. The 2 experimental periods lasted 14 d and were separated by a 7-d washout interval. Samples were collected on d 0, 13, and 14 of each period. Data from d 0 were used as covariate. Significance was declared at P ≤ 0.05 and tendency at 0.05 < P ≤ 0.10. There was a 10-fold increase in the relative concentration of bacteria from the Bacillus subtilis group in the rumen when feeding direct-fed Bacillus compared with control. Treatment did not affect ruminal pH, NH₃-N, or concentrations of acetate, propionate, and butyrate. However, direct-fed Bacillus increased ruminal concentrations of isovalerate and isobutyrate (tendency). Treatments did not affect lactation performance. Supplying direct-fed Bacillus enhanced milk relative concentration of anteiso 13:0 by 27.3% and of anteiso 15:0 by 6.5% and tended to increase concentrations of iso 14:0 (+41.8%) relative to control. When expressed on a yield basis, direct-fed Bacillus increased the secretion of anteiso 13:0 and decreased that of 11:0, 15:0, 17:0 (tendency), and cis-9 17:1. These variations, although limited in magnitude, indicate that milk branched-chain fatty acid composition is sensitive to ruminal microbiota modifications without changes in chemical composition of the diet.     

Metabolic fate of long-chain unsaturated fatty acids and their effects on palmitic acid metabolism and gluconeogenesis in bovine hepatocytes

The objectives were to determine the metabolic fate of different long-chain fatty acids, and their effects on palmitic acid metabolism and gluconeogenesis in bovine hepatocytes. Hepatocytes were isolated from four ruminating calves and exposed in suspension for 3 h to one of the following treatments: 1 mM palmitic acid (1C16), 2 mM palmitic acid (2C16), or 1 mM palmitic acid plus either 1 mM oleic (C18:1), linoleic (C18:2), linolenic (C18:3), eicosapentaenoic (C20:5), or docosahexaenoic acid (C22:6). Oxidation of [1-(14)C]palmitic acid or one of the [1-(14)C]-labeled treatment fatty acids to CO2 or incorporation into cellular triglycerides (TG), phospholipids, cholesterol, and cholesterol esters were measured. Rates of oxidation to CO2 were 3- to 4-fold higher for C22:6 than for other fatty acids, with the exception of C20:5, which had intermediate rates of oxidation to CO2. In general, treatments 2C16 and C18:1 yielded the highest rates of incorporation into most cellular lipids, whereas the polyunsaturated fatty acids were poor substrates for incorporation into cellular lipids. The most pronounced change was a large reduction of polyunsaturated fatty acid incorporation into cellular TG compared to 1C16, 2C16, and C18:1. The unsaturated fatty acids also influenced palmitic acid metabolism. The addition of C20:5 yielded the highest rates of palmitic acid oxidation to CO2 followed by addition of C18:1 and C22:6. Treatments containing polyunsaturated fatty acids decreased palmitic acid metabolism to TG and total cellular lipids compared with treatments 2C16 and C18:1. Rates of gluconeogenesis from propionate were significantly higher for the treatment containing C18:1. Long-chain fatty acids vary in their routes of metabolism and influence palmitic acid metabolism and gluconeogenesis in bovine hepatocytes.     

Effect of sources of calcium salts of fatty acids on production,nutrient digestibility,energy balance,and carryover effects of early lactation grazing dairy cows

The objective of our study was to investigate the effects of sources of calcium salts of fatty acids (FA) on production, nutrient digestibility, energy balance, and carryover effects of early lactation grazing dairy cows. Treatment diets were offered from 3 to 16 wk postpartum (the treatment period), in which all cows grazed elephantgrass (Pennisetum purpureum ‘Cameroon’) and treatments were added to a concentrate supplement. The treatments were (1) control (concentrate without supplemental fat); (2) concentrate with calcium salts of soybean FA (CSSO); and (3) concentrate with calcium salts of palm FA (CSPO). From 17 to 42 wk postpartum (the carryover period), all cows received a common diet fed as a total mixed ration. During the treatment period, CSPO increased milk yield, milk fat yield, 3.5% fat-corrected milk, energy-corrected milk, and cumulative milk yield compared with control and CSSO. Treatment CSSO increased the yield of milk but did not affect 3.5% fat-corrected milk or energy-corrected compared with control. Also, CSSO decreased milk fat yield, dry matter intake, neutral detergent fiber digestibility, and body weight and body condition loss. Compared with control, both CSSO and CSPO increased feed efficiency (3.5% fat-corrected milk:dry matter intake), and CSPO increased feed efficiency compared with CSSO. When considering energy partitioning (as % energy intake), CSPO increased energy partitioning toward milk and increased energy mobilized from body reserves compared with control and CSSO. Furthermore, CSSO tended to reduce the mobilization of energy from body reserves compared with control. In the carryover period, no differences in milk composition were observed among treatments. A treatment by time interaction was observed during the carryover period for milk yield because cows on CSPO maintained higher production compared with control and CSSO cows until 30 wk postpartum; CSSO had a lower carryover effect sustaining higher milk yield compared with control until 25 wk postpartum. In conclusion, supplementation with CSPO was an effective strategy to increase energy intake and yields of milk and milk solids and it had a greater carryover effect. Supplementation with CSSO resulted in lower mobilization of reserves and less variation in body weight and body condition throughout lactation.     

Interaction of probiotic Lactobacillus and Bifidobacterium strains with human intestinal epithelial cells: adhesion properties, competition against enteropathogens and modulation of IL-8 production

The human intestinal microbiota plays a pivotal role in human nutrition and health by promoting the supply of nutrients, preventing pathogen colonization and shaping and maintaining normal mucosal immunity. The depletion of the individual microbiota can result in a higher susceptibility to enteropathogenic bacteria infection. In order to reduce this risk, the use of food supplements containing probiotic bacteria has been recently addressed. In this paper, we investigate the protective role toward enteropathogen infection of probiotic strains belonging to Lactobacillus and Bifidobacterium. According to our experimental data, Lactobacillus acidophilus Bar13, L. plantarum Bar10, Bifidobacterium longum Bar33 and B. lactis Bar30 were effective in displacing the enteropathogens Salmonella typhimurium and Escherichia coli H10407 from a Caco-2 cell layer. Moreover, L. acidophilus Bar13 and B. longum Bar33 have been assessed for their immunomodulatory activity on IL-8 production by HT29 cells. Both strains showed the potential to protect enterocytes from an acute inflammatory response. These probiotic strains are potential candidates for the development of new functional foods helpful in counteracting enteropathogen infections.     

Effect of the feeding system on intramuscular fatty acids and conjugated linoleic acid isomers of beef cattle,with emphasis on their nutritional value and discriminatory ability

Thirty two Alentejano purebred bulls were used to investigate the effect of four feeding systems (pasture only, pasture feeding followed by 2 or 4 months of finishing on concentrate, and concentrate only) on meat fatty acid composition (GC-FID), including conjugated linoleic acid (CLA) isomeric distribution (Ag[+]–HPLC–DAD). In addition, meat fatty acids and CLA isomers were used to elucidate the impact of the different feeding regimens on the nutritional value of intramuscular fat and their usefulness as chemical discriminators of meat origin. The diet had a major impact on the fatty acid composition of beef (affected 27 of 36 fatty acids and 10 of 14 CLA isomers), which was independent of the fatty acid concentration. Beef fat from pasture-fed animals had a higher nutritional quality relative to that from concentrate-fed bulls. Finally, meat fatty acid composition was an effective parameter to discriminate between ruminant feeding systems, including different finishing periods on concentrate.     

Corn grain-processing method interacts with calcium salts of palm fatty acids supplementation on milk production and energy balance of early-lactation cows grazing tropical pasture

The objective of our study was to investigate the associative effects of feeding Ca salts of palm fatty acids (FA) and corn grain-processing method on production, nutrient digestibility, energy balance, and carryover effects of early-lactation dairy cows grazing a tropical pasture. Treatment diets were offered from 3 to 16 wk postpartum (treatment period), in which all cows grazed elephant grass (Pennisetum purpureum L. Cameroon) and treatments were added to a concentrate supplement. Treatments were flint corn grain-processing method either as fine ground (FGC) or steam-flaked (SFC) associated with Ca salts of palm FA supplementation either not supplemented or supplemented (CSPO). From 17 to 40 wk postpartum (carryover period) all cows received a common diet fed as total mixed ration. During the treatment period, a tendency for an interaction between CSPO and corn grain-processing method were observed for milk yield, milk fat yield, and energy-corrected milk (ECM), as CSPO caused them to increased to a greater extent in the FGC diet compared with the SFC diet. Furthermore, a tendency for an interaction between CSPO and corn grain-processing method was observed for body weight change, because CSPO increased body weight loss in the FGC diet but not in the SFC diet. The CSPO increased milk yield, milk fat yield, 3.5% fat-corrected milk, ECM, and cumulative milk yield compared with not supplemented. Also, CSPO increased energy intake, milk energy output, and energy partitioning toward milk, whereas reduced energy was allocated to body reserves. The SFC increased milk yield, ECM, milk protein yield, milk casein yield, and cumulative milk yield, and decreased milk urea N compared with FGC. The SFC compared with FGC also increased body condition score and body weight change, and increased energy partitioning toward body reserves. During the carryover period, an interaction between CSPO and corn grain-processing method was observed for milk yield, which occurred because CSPO maintained higher milk yield in the FGC diet but not in the SFC diet. Therefore, in the carryover period, the additive effect between SFC and CSPO that occurred in the treatment period was not maintained throughout the carryover period. However, CSPO increased yields of milk fat, protein, casein as well as fat-corrected milk and ECM. In conclusion, corn grain-processing method interacts with CSPO supplementation on production responses and carryover effects of grazing cows. When CSPO was fed in the FGC diet, milk production increased to a greater extent than when fed in the SFC diet, but also caused greater mobilization of reserves at early lactation. This suggests an interaction between fat supplementation and corn grain-processing method on energy partitioning of dairy cows. Also, both supplementation with CSPO and SFC were effective strategies to increase energy intake and yields of milk and milk solids. The carryover effect on milk production was greater for CSPO supplementation than corn grain-processing method, whereas feeding SFC diets had lower mobilization of reserves and less body weight and body condition score variation throughout lactation.     

Differential effects of n-3 and n-6 fatty acids on prostaglandin F2alpha production by bovine endometrial cells

Recent studies have implicated n-3 polyunsaturated fatty acids in the reduction of eicosanoid production in the bovine uterus. The objective of this study was to determine whether the effect of eicosapentaenoic acid (EPA; C_20:5, n-3) on PGF_2α production by bovine endometrial (BEND) cells is influenced by the quantity of linoleic acid (C_18:2, n-6) in the incubation medium. Confluent BEND cells were incubated in the absence (control) or presence of 100 μMof EPA for 24 h. After incubation, cells were rinsed and then stimulated with phorbol 12,13-dibutyrate (PDBu; 100 ng/mL) for 6 h. Additional sets of culture dishes were treated with a combination of EPA and increasing n-6/n-3 fatty acid ratios for 24 h and then challenged with PDBu for 6 h. The PDBu stimulated PGF_2α secretion and upregulated steady-state concentrations of prostaglandin endoperoxide synthase-2 and peroxisome proliferator-activated receptor delta mRNA within 6 h. Preincubation of BEND cells with EPA for 24 h decreased PGF_2α response to phorbol ester, but had no detectable effects on prostaglandin endoperoxide synthase-2 or peroxisome proliferator-activated receptor delta mRNA abundance in PDBu-stimulated BEND cells. The inhibitory effect of EPA on PGF_2α production was reverted in BEND cells treated with an increasing n-6-to-n-3 fatty acid ratio. Findings indicate that the net inhibition of endometrial PGF_2α bioynthesis by n-3 fatty acids may vary depending on the ratio of n-6 to n-3 fatty acids in the uterus.     

The cytotoxic effect of Bowman–Birk isoinhibitors,IBB1 and IBBD2, from soybean (Glycine max) on HT29 human colorectal cancer cells is related to their intrinsic ability to inhibit serine proteases

Bowman–Birk inhibitors (BBI) from soybean and related proteins are naturally occurring protease inhibitors with potential health‐promoting properties within the gastrointestinal tract. In this work, we have investigated the effects of soybean BBI proteins on HT29 colon adenocarcinoma cells, compared with non‐malignant colonic fibroblast CCD‐18Co cells. Two major soybean isoinhibitors, IBB1 and IBBD2, showing considerable amino acid sequence divergence within their inhibitory domains, were purified in order to examine their functional properties, including their individual effects on the proliferation of HT29 colon cancer cells. IBB1 inhibited both trypsin and chymotrypsin whereas IBBD2 inhibited trypsin only. Despite showing significant differences in their enzyme inhibitory properties, the median inhibitory concentration values determined for IBB1 and IBBD2 on HT29 cell growth were not significantly different (39.9±2.3 and 48.3±3.5 μM, respectively). The cell cycle distribution pattern of HT29 colon cancer cells was affected by BBI treatment in a dose‐dependent manner, with cells becoming blocked in the G0–G1 phase. Chemically inactive soybean BBI had a weak but non‐significant effect on the proliferation of HT29 cells. The anti‐proliferative properties of BBI isoinhibitors from soybean reveal that both trypsin‐ and chymotrypsin‐like proteases involved in carcinogenesis should be considered as potential targets of BBI‐like proteins.     

The effect of feed supplementation with dietary sources of n‐3 polyunsaturated fatty acids,flaxseed and algae Schizochytrium sp., on their incorporation into lipid fractions of Japanese quail eggs

The influence of feed supplemented with dietary sources of n‐3 polyunsaturated fatty acids (PUFAs), namely flaxseed and algae Schizochytrium sp., on the fatty acid composition of lipids from Japanese quail eggs was studied. Two groups of quails were fed for 19 weeks, starting from 6th week of life. Lipid fractions from dried egg mass of total of 4300 eggs of control and experimental group were analysed. The fatty acid profile was affected in both triacylglycerols and phospholipids. The significant (P < 0.05) increase in n‐3 PUFAs up to 7.33% in the total lipids, α‐linolenic acid (ALA, 18:3) up to 4.57% in triacylglycerols, docosahexaenoic acid (22:6, DHA) up to 23.91% in phosphatidylethanolamine and up to 4.35% in phosphatidylcholine as well as the significant (P < 0.05) reduction in n‐6/n‐3 ratio in the lipid fractions to the range of 0.9–3.7 was observed in experimental group. The incorporation of different n‐3 into lipids was selective: ALA was found in the triacylglycerol fraction, whereas EPA, DPA and DHA were detected in the phospholipid fractions, exclusively at sn‐2 position. The studies showed that the feeding quails with modified diets is effective way of biofortifying the eggs with nutritionally desired n‐3 PUFAs.