Difference in in vitro fermentability of four carbohydrates and two diets,using ileal and faecal inocula from unweaned piglets

An experiment was conducted to examine differences in the in vitro fermentability of four carbohydrate‐rich feed ingredients and two weaning piglet diets with and without these ingredients, using both the ileal contents and the faeces of unweaned piglets as inocula. In the first part of the experiment, cumulative gas production was measured over time, using faecal inocula mixed from nine specially raised crossbred piglets (no creep feed or antibiotics) at 3 weeks of age. Inulin, lactulose, unmolassed sugar beet pulp, wheat starch and the two diets were used as substrates and fermented in vitro for 72 h. Gas production was measured as an indicator of the kinetics of fermentation. Fermentation end‐products, including volatile fatty acids and ammonia, and organic matter loss, were also measured. For the fermentations of feed ingredients, samples were also collected for polymerase chain reaction/denaturing gradient gel electrophoresis analyses initially and after the fermentation process, to study changes in the composition of the bacterial community. This procedure was repeated 1 week later, using ileal contents from the same piglets as inoculum. There were significant differences between the inocula, in terms of both overall fermentation characteristics and composition and between the substrates. There was also a significant interaction between inocula and substrates, which suggests that there were potentially important differences in the microbial activity occurring in these two areas of the gastrointestinal tract (GIT). For the two diets, one with and one without addition of these fermentable ingredients, there were significant differences in terms of the kinetics, but less so in terms of the end‐products of fermentation. It was concluded that inocula from both the small and large intestine should be used to obtain a more accurate assessment of potential feed ingredients which will stimulate fermentation in the piglet GIT. Copyright © 2005 Society of Chemical Industry     

In vitro faecal fermentation outcomes and microbiota shifts of resistant starch spherulites

In the present study, we prepared A- and B-type polymorph-resistant starch spherulites (RSSs) with a wide range of crystallinity, and elucidated the relation between crystalline structure and faecal microbial fermentation outcomes. A- and B-type polymorphic RSSs displayed similar fermentation rate throughout entire process with final gas production of 11.8–13.2 mL, suggesting that the crystalline pattern and crystallinity cannot effectively control the fermentation rate of starches. The acetate and butyrate concentration of A-type RSS decreased with the increase of crystallinity respectively, and highly crystalline B-type RSS reduced the production of acetate (8.4 mM) and propionate (2.1 mM). RSSs with different crystalline types showed similar microbial community structure but distinct key bacteria species. A-type RSSs greatly promoted the abundance of butyrogenic bacteria (i.e., Roseburia faecis and Lachnospiraceae), whereas B-type RSSs tended to proliferate Prevotella copri associated with propionate production. Our findings shed new light on design and manufacture of starch-based functional ingredients with enhanced gut health.     

Effect of total mixed ration composition on fermentation and efficiency of ruminal microbial crude protein synthesis in vitro

The goal of this study was to identify dietary factors that affect fermentation and efficiency of microbial crude protein (CP_M) synthesis in the rumen in vitro. We used 16 total mixed, dairy cow rations with known digestibilities that varied in ingredient composition and nutrient content. Each ration was incubated in a Rusitec (n = 3) for 15 d, and fermentation of different fractions was assessed. Observed extents of fermentation in 24 h were 35 to 47% for organic matter, 25 to 60% for crude protein, 3 to 28% for neutral detergent fiber, and 31 to 45% for gross energy. Organic matter fermentation depended on the content of crude protein and neutral detergent fiber in the ration. We studied net synthesis of CP_M using an ¹⁵N dilution technique and found that 7 d of continuous ¹⁵N application are needed to achieve an ¹⁵N enrichment plateau in the N of isolated microbes in this type of study. The efficiency of CP_M synthesis was 141 to 286 g/kg of fermented organic matter or 4.9 to 11.1 g/MJ of metabolizable energy, and these ranges agree with those found in the literature. Multiple regressions to predict the efficiency of CP_M synthesis by diet data showed that crude protein was the only dietary chemical fraction that had a significant effect. Fat content and the inclusion rate of corn silage in the ration also tended to improve efficiency. We suggest that microbial need for preformed amino acids may explain the crude protein effect. A large part of the variation in efficiency of microbial activity still remains unexplained.     

辣椒素对肠道菌体外发酵的影响

食用辣椒会带来肠道不适反应,对肠道环境存在潜在影响。实验拟通过体外发酵验证辣椒素对肠道发酵环境的影响。向大鼠盲肠内容物发酵液中添加0.05%、0.10%、0.15%三种浓度辣椒素,培养10h后检测发酵液中pH、游离氨、短链脂肪酸(SCFA)及双歧杆菌、乳酸杆菌、大肠杆菌3种微生物的变化趋势,探讨辣椒素对肠道发酵环境的影响。结果表明,与空白组相比,添加辣椒素使盲肠内容物发酵液pH、游离氨浓度升高,短链脂肪酸含量降低,双歧杆菌、乳酸杆菌数量下降,而大肠杆菌数量上升。所有变化均与辣椒素添加剂量存在相关性。研究结果表明,辣椒素对肠道发酵环境有潜在危害。      

A comparison between buffalo (Bubalus bubalis) and cow (Bos taurus) rumen fluids in terms of the in vitro fermentation characteristics of three fibrous feedstuffs

Rumen fluids from fistulated buffalos (Italy‐BRF) and cows (Netherlands‐CRF) were used as inocula to determine the fermentation kinetics of three forages. These were corn silage (CS), grass silage (GS) and wheat straw (WS) which had originated from both regions, giving six substrates in total. Fermentation kinetics was assessed by the measurement of cumulative gas production. Organic matter (OM) loss and volatile fatty acid (VFA) concentration at the end of the fermentation period were also determined. Both BRF and CRF ranked the substrates in the same order for total VFA, total gas production and the maximum rate of substrate degradation (CS > GS > WS). However, while the ranking of substrates was the same for both species, the absolute values differed significantly between the two inocula. Gas production, expressed as cumulative volume per unit mass of incubated (OMCV, ml g^?1) and as cumulative volume per unit mass of OM degraded (OM ml g^?1) for CRF was consistently higher than that for BRF for all substrates (p < 0.0001). VFA production, particularly of acetic and butyric acids, was significantly (p < 0.0001) lower for BRF than for CRF, though the digestibility of OM was the same for both inocula. VFA production predicted by the use of stoichiometric equations was generally higher than the observed gas production. These equations also predicted that the calculated amount of OM utilised for microbial growth was higher for BRF than for CRF. This may explain why diets having the same energy content but less protein can be fed to buffalo since they seem to have a lower requirement for protein compared with cattle. This possibility will need to be investigated in vivo. Given the similar ranking of feedstuffs between the two sources of inocula, these results suggest that either inoculum would be suitable for use in the cumulative gas production test as a measure of feedstuff evaluation. However, given the differences in absolute values, it is recommended that inocula from the species which will receive the feedstuff should be used. Copyright © 2004 Society of Chemical Industry     

肠道微生物体外发酵模型研究进展及其在食品中的应用

随着分子微生态学,特别是高通量测序技术的快速发展,人们对肠道微生物的作用有了进一步认识。肠道不仅是消化吸收的场所,其微生物菌群现更被认为是一个高度特异化的\     

微生物发酵改善菜籽粕品质的研究

采用实验室筛选的菌种混合固态发酵菜籽粕,比较菜籽粕发酵前后硫甙、粗蛋白质、菜籽肽、乳酸含量以及蛋白质体外消化率的变化.结果表明,菜籽粕中的硫甙降解率为53.4%,粗蛋白质含量从39.99%提高到45.84%,菜籽肽含量由2.48%提高到10.58%,氨基酸的组成和含量也有一定的变化和提高,蛋白质的快速体外消化率提高了1.94%,有益代谢产物乳酸的含量由1.03%提高到2.53%.     

Effects of Euphorbia esula L. (leafy spurge) on cattle and sheep in vitro fermentation and gas production

BACKGROUND: Euphorbia esula L. (leafy spurge) is indigenous to Eurasia and has been known to cause grazing aversion in ruminant species. As a result, E. esula encroachment has negatively impacted rangelands in the Northern Great Plains and Intermountain West of the USA, as well as southern Canada. Our objectives were to evaluate the effect of increasing concentrations of E. esula on in vitro dry matter digestibility (DMD) and gas production. Two ruminally‐cannulated cows and ewes were used as rumen inoculum donors. To accomplish objectives, two studies were conducted. In study 1, animals were fed exclusively a barley hay (12% crude protein (CP), 55.4% neutral detergent fiber (NDF), DM basis) diet; whereas in study 2, animals were fed a diet of 15% E. esula (21.9% CP, 48% NDF, DM basis) and 85% barley hay based on previous day intake. RESULTS: The 24 and 48 h in vitro and 96 h gas production indicate that, regardless of inoculum source or substrate fermented, DMD was not influenced. Differences, however, were consistently observed across studies for NDF disappearance. CONCLUSION: Regardless of inoculum source NDF disappearance was greater when substrate being fermented contained 0%, 80%, or 100% E. esula. Published 2011 by John Wiley & Sons, Ltd.     

柿子单宁对体外模拟发酵环境中肠道微生物的影响

采用体外厌氧发酵法研究了柿子单宁对肠道微生物的影响。将单宁与大鼠肠道微生物共孵育,分别测定0、6、12、24、48 h五个时间点发酵液中六种微生物、短链脂肪酸和p H的变化。结果显示,与空白组相比,添加2 g/L剂量组的柿单宁能有效地促进肠道有益菌(乳杆菌和双歧杆菌)的增殖,抑制有害菌(肠球菌、大肠杆菌、梭菌和拟杆菌)的增殖,增加短链脂肪酸含量,降低发酵液p H。研究结果表明柿子单宁对肠道微生态具有调节作用,可能是其在体内发挥作用的机制之一。      

Unique interrelationships between fiber composition,water-soluble carbohydrates,and in vitro gas production for fall-grown oat forages

Sixty samples of ‘ForagePlus’ oat were selected from a previous plot study for analysis of in vitro gas production (IVGP) on the basis of 2 factors: (1) high (n = 29) or low (n = 31) neutral detergent fiber (NDF; 62.7 ± 2.61 and 45.1 ± 3.91%, respectively); and (2) the range of water-soluble carbohydrates (WSC) within the high- and low-NDF groups. For the WSC selection factor, concentrations ranged from 4.7 to 13.4% (mean = 7.9 ± 2.06%) and from 3.5 to 19.4% (mean = 9.7 ± 4.57%) within high- and low-NDF forages, respectively. Our objectives were to assess the relationships between IVGP and various agronomic or nutritional characteristics for high- and low-NDF fall-oat forages. Cumulative IVGP was fitted to a single-pool nonlinear regression model: Y = MAX × (1 – e ^{[–K× (t – lag)]}), where Y = cumulative gas produced (mL), MAX = maximum cumulative gas produced with infinite incubation time (mL), K = rate constant, t = incubation time (h), and lag = discrete lag time (h). Generally, cumulative IVGP after 12, 24, 36, or 48 h within high-NDF fall-oat forages was negatively correlated with NDF, hemicellulose, lignin, and ash, but positively correlated with WSC, nonfiber carbohydrate (NFC), and total digestible nutrients (TDN). For low-NDF fall-grown oat forages, IVGP was positively correlated with growth stage, canopy height, WSC, NFC, and TDN; negative correlations were observed with ash and crude protein (CP) but not generally with fiber components. These responses were also reflected in multiple regression analysis for high- and low-NDF forages. After 12, 24, or 36 h of incubation, cumulative IVGP within high-NDF fall-oat forages was explained by complex regression equations utilizing (lignin:NDF)², lignin:NDF, hemicellulose, lignin, and TDN² as independent variables (R² ≥ 0.43). Within low-NDF fall-grown oat forages, cumulative IVGP at these incubation intervals was explained by positive linear relationships with NFC that also exhibited high coefficients of determination (R² ≥ 0.75). Gas production was accelerated at early incubation times within low-NDF forages, specifically in response to large pools of WSC that were most likely to be present as forages approached boot stage by late-fall.     

Different concentrations of grape seed extract affect in vitro starch fermentation by porcine small and large intestinal inocula

BACKGROUND: Grape seed extract (GSE) phenolics have potential health‐promoting properties, either from compounds present within the extract, or metabolites resulting from gastrointestinal tract (GIT) fermentation of these compounds. This study describes how GSE affected the kinetics and end‐products of starch fermentation in vitro using pig intestinal and fecal inocula. Six GSE concentrations (0, 60, 125, 250, 500, and 750 µg ml^?1 were fermented in vitro by porcine ileal and fecal microbiota using starch as the energy source. Cumulative gas production, and end‐point short chain fatty acids and ammonia were measured. RESULTS: GSE phenolics altered the pattern (gas kinetics, and end‐products such as SCFA and NH ) of starch fermentation by both inocula, at concentrations above 250 µg ml^?1. Below this level, neither inoculum showed any significant (P > 0.05) effect of the GSE. CONCLUSION: The results show that GSE phenolics at a concentration over 250 µg ml^?1 can have measurable effects on microbial activity in an in vitro fermentation system, as evidenced by the changes in kinetics and end‐products from starch fermentation. This suggests that fermentation patterns could be conceivably shifted in the actual GIT, though further evidence will be required from in vivo studies. Copyright © 2012 Society of Chemical Industry     

Effects of nonstructural carbohydrate concentration in alfalfa on fermentation and microbial protein synthesis in continuous culture

Insufficient readily fermentable energy combined with extensive degradation of proteins in alfalfa (Medicago sativa L.) may result in poor forage N utilization by ruminants. Using the inherent genetic variability and differences between harvests, our objective was to compare the effect of contrasting concentrations of nonstructural carbohydrates (NSC) in alfalfa on rumen fermentation and microbial protein synthesis. Individual genotypes of the alfalfa cultivar AC Caribou grown near Québec City, Québec, Canada, were harvested at the vegetative and early flowering stages, dried at 55°C, ground, and analyzed for soluble carbohydrates (fructose + sucrose + glucose + pinitol) and starch. Approximately 20 genotypes having, respectively, the highest and lowest NSC concentrations were pooled to constitute 2 contrasted 1-kg forage samples. Samples of high- (17.9% DM) and low- (7.4% DM) NSC alfalfa were respectively allocated to separate dual-flow fermenters in a completely randomized design with 3 replications. Rumen inoculum was obtained from 4 ruminally fistulated cows in early lactation that were fed a TMR with a 50:50 forage to concentrate ratio. A 10-d incubation period was used, with the first 6 d serving as an adaptation period followed by 4 d of sampling with solid and liquid dilution rates in the fermenters set at approximately 2.0 and 4.3%/h, respectively. High versus low NSC concentration in alfalfa significantly enhanced the apparent digestibility of OM (59.1% for high-NSC alfalfa vs. 54.4% for low-NSC alfalfa) and DM (60.0 vs. 54.3%) and the true digestibility of DM (74.1 vs. 64.7%). Increasing NSC concentration in alfalfa (high vs. low) significantly decreased ruminal pH (6.85 vs. 7.08) and NH₃-N concentration (26.0 vs. 33.6 mg/dL) and increased total VFA concentration (94.9 vs. 83.0 mM). Molar proportions of acetate, isobutyrate, and isovalerate significantly decreased, whereas molar proportions of propionate and butyrate significantly increased with high-NSC alfalfa, resulting in a more glucogenic fermentation. More importantly, microbial N flow (263 vs. 230 mg/d) and bacterial N efficiency (41.1 vs. 29.6% of available N), measured using ¹⁵N as a microbial marker, both significantly increased with the high-NSC alfalfa. These results indicate that increasing the concentration of NSC in alfalfa promotes a glucogenic fermentation and enhances microbial N synthesis in the rumen.     

The effect of betaine on in vitro fermentation of carbohydrate and protein combinations under osmotic stress in pigs

BACKGROUND: There is evidence that dietary betaine might improve nutrient digestibilities in pigs due to its osmoprotective properties. This in vitro study was performed to assess the potential of supplemented betaine sources to affect fermentation characteristics of different carbohydrate and protein combinations, under conditions of osmotic stress. In vitro fermentation was performed by use of the modified Hohenheim gas test with pig faeces as microbial inoculum. RESULTS: At the end of in vitro fermentation under osmotic stress conditions, differences (P < 0.05) in gas production, ammonia concentration, and concentration of short‐chain fatty acids were observed for the different carbohydrates, while for the two proteins only differences (P < 0.05) in ammonia and short‐chain fatty acid concentration were obtained. Supplementation of a native betaine source rather than addition of pure betaine increased propionic acid (P < 0.05) and butyric acid (P < 0.05) production. CONCLUSION: These results suggest that only supplementation of a native betaine source but not of pure betaine products affects microbial fermentation under osmotic stress. Furthermore, both the carbohydrate and protein source play an important role in modifying microbial activity under osmotic stress conditions. It can also be concluded that the modified Hohenheim gas test is suitable to scrutinise fermentation activity of different assay substrates under osmotic stress. Copyright © 2012 Society of Chemical Industry     

Processing of rye bran influences both the fermentation of dietary fibre and the bioconversion of lignans by human faecal flora in vitro

Plant lignans are converted to mammalian forms, enterodiol and enterolactone, in the colon. Enhanced plasma or urinary enterolactone levels have been associated with lowered risk of cardiovascular diseases and breast cancer. Processed rye bran and its fractions were compared to ascertain the fermentation rates of fermentable carbohydrates and the bioconversion of lignans. Rye bran was extruded and treated with a food‐grade xylanase. Part of the xylanase‐treated rye bran was separated into a soluble rye bran extract and an insoluble residue, and a part of the xylanase‐treated rye bran was freeze‐dried without separation. All the samples were digested by pepsin and pancreatin and subsequently fermented with a human faecal inoculum in vitro. The consumption of carbohydrates, the productions of short‐chain fatty acids (SCFA), enterodiol and enterolactone were followed as a function of time. The soluble rye bran extract showed the fastest fermentation rate and the highest extent of fermentation determined as the consumption of neutral sugar residues (arabinose, xylose and glucose), the production of SCFA and the formation of enterodiol and enterolactone. Xylanase treatment enhanced the fermentation rate of extruded rye bran. An even a higher fermentation rate was observed for rye bran extract containing soluble carbohydrates. The amount of enterolactone precursors in rye seemed to be too low for enterolactone formation using an amount of substrate suitable for carbohydrate fermentation. However, xylanase treatment enhanced the availability of plant lignans from rye bran, as enterodiol formation was increased by the use of xylanase. Copyright © 2005 Society of Chemical Industry     

微生物发酵对豆粕抗原性的影响

探讨了微生物发酵对豆粕抗原性的影响。选用植物乳杆菌、干酪乳杆菌、枯草芽孢杆菌、地衣芽孢杆菌和米曲霉这5种菌株,在液态和固态条件下分别发酵豆粕12 h,对发酵产物进行抗原性测定。结果表明:豆粕经这5种菌株发酵后,粗蛋白含量均有所提高,其中枯草芽孢杆菌在固态发酵时降解豆粕抗原蛋白和降低豆粕抗原性的效果优于其它菌株,此时,豆粕蛋白水解度为4.89%,必需氨基酸含量为193.51mg/g。SDS-PAGE显示发酵豆粕中β-伴大豆球蛋白的α’和α亚基消失,β亚基条带和大豆球蛋白的酸性亚基条带密度减弱,同时大豆球蛋白与β-伴大豆球蛋白的抗原性降低率分别为20.62%和50.12%。     

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.     

Effects of forage particle size,forage source,and grain fermentability on performance and ruminal pH in midlactation cows

Our study investigated the effects of, and interactions between, forage particle size, level of dietary ruminally fermentable carbohydrate (RFC), and level of dietary starch on performance, chewing activity, and ruminal pH for dairy cows fed one level of dietary NDF. Twelve cows (48 DIM) were assigned to six treatments in a replicated 6 x 6 Latin square. Treatments were arranged in an incomplete 2 x 2 x 2 factorial design. Factors were: dry cracked shelled corn (DC, low RFC) or ground high-moisture corn (HMC; high RFC), finely chopped or coarse silage, and alfalfa silage as the only forage or a 50:50 ratio (DM basis) of alfalfa and corn silage. Diets combining HMC with only alfalfa silage were not included in the experiment. Diets were fed for ad libitum intake as a TMR with a concentrate:forage ratio of 61:39. Diets based on only alfalfa silage and diets based on a mix of alfalfa and corn silage averaged 18.6 and 15.8% CP, 25.8 and 24.7% NDF, 17.7 and 14.8% ADF, and 29.1 and 37.3% starch, respectively. Mean particle sizes were 5.3, 2.7, 5.6, and 2.8 mm for coarse alfalfa, fine alfalfa, coarse corn silage, and fine corn silage, respectively. Decreasing forage particle size decreased DMI (23.3 vs. 21.6 kg) and organic matter intake (22.0 vs. 20.2 kg). Increasing RFC decreased DMI (22.8 vs. 21.0 kg) and organic matter intake (21.5 vs. 20.0 kg). Decreasing forage particle size increased energy-corrected milk for alfalfa based diets (34.9 vs. 37.4 kg). Percentage of milk fat decreased with decreasing forage particle size (3.07 vs. 2.90%) and increased level of RFC (3.04 vs. 2.57%). Percentage of protein increased when corn silage partially replaced alfalfa silage (2.84 vs. 2.90%) but decreased when HMC replaced DC (2.90 vs. 2.84%). Apparent total tract digestibility of DM (66.7 vs. 68.5%), OM (65.9 vs. 70.7%), and starch (88.9 vs. 93.4%) increased when level of RFC was increased. Increasing level of RFC decreased mean ruminal pH from 5.82 to 5.67 and decreased minimum pH. Hours per day at which pH was <5.8, and area <5.8, increased when corn silage partially replaced alfalfa silage (2.6 vs. 4.4 h and 8.9 h x pH vs. 11.4 h x pH) and decreased further when level of RFC was increased (4.4 vs. 6.4 h and 11.4 h x pH vs. 14.3 h x pH). Decreasing forage particle size in HMC diets increased hours and area <5.8, but for DC diets, the effect of forage particle size depended on forage source. Interactions were found between level of physically effective fiber, forage source, and level of RFC on production and pH, complicating the inclusion of these effects in dairy ration formulation and evaluation.