乳杆菌胞外多糖对人体粪便菌群及其产生短链脂肪酸的影响

目的在体外利用7名志愿者的新鲜粪便作为来源,选择菊粉作为对照,研究干酪乳杆菌LC2W与鼠李糖乳杆菌Y37胞外多糖对人体肠道菌群的影响。方法利用PCR-DGGE(变性梯度凝胶电泳)方法分析样品中菌群组成,采用高效液相色谱的方法分析短链脂肪酸组成。结果与菊粉相比,粪便菌群在添加乳杆菌胞外多糖培养后,可以促进短链脂肪酸的生成,保持肠道菌群的多样性。结论干酪乳杆菌LC2W与鼠李糖乳杆菌Y37的胞外多糖在体外有调节粪便菌群的作用。     

Chemical specificity in short-chain fatty acids and their analogues in increasing osmotic fragility in rat erythrocytes in vitro

We examined the role of the chemical specificity of short-chain fatty acids (SCFAs) and their derivatives in increasing osmotic fragility (OF) in rat red blood cells (RBCs). Except for formic acid, normal SCFAs with 2 to 8 carbons increased the OF in rat RBCs with increasing number of hydrocarbons in a dose-dependent manner. Replacement of the carboxylic group with sulfonic group inhibited, but did not abolish, the SCFA-mediated increase in OF. Introduction of another carboxylic group (dicarboxylic acids) completely abolished the SCFA-mediated increase in OF. Transformation of the hydrocarbon chains in SCFAs from straight to branched or cyclic chains affected the degree of the OF-increasing effect. Introduction of double- or triple-carbon bonds to the hydrocarbon chain in parent SCFAs did not affect the increase in OF. Both hydrophilic (carboxylic group) and hydrophobic elements (hydrocarbons) at opposite sides of a molecule were required to affect the RBC membrane, and the size and form of hydrophobic element were important factors in determining the SCFA-mediated increase in OF. The hydrocarbon chains probably enter the plasma membrane, with the hydrophilic carboxylic base remaining outside of the membrane, and interact with phospholipid in cell membrane and disturb the structure of lipid layer resulting in the increase in OF in the rat RBCs.     

Permeability properties of apical and basolateral membranes of the guinea pig caecal and colonic epithelia for short-chain fatty acids

Unidirectional fluxes of short-chain fatty acids (SCFA) indicated marked segmental differences in the permeability of apical and basolateral membranes. The aim of our study was to prove these differences in membrane permeability for a lipid-soluble substance and to understand the factors affecting these differences. Apical and basolateral membrane fractions from guinea pig caecal and colonic epithelia were isolated. Membrane compositions were determined and the permeability of membrane vesicles for the protonated SCFA was measured in a stopped-flow device. Native vesicles from apical membranes of the caecum and proximal colon have a much lower permeability than the corresponding vesicles from the basolateral membranes. For the distal colon, membrane permeabilities of native apical and basolateral vesicles are similar. In vesicles prepared from lipid extracts, the permeabilities for the protonated SCFA are negatively correlated to cholesterol content, whereas no such correlation was observed in native vesicles. Our findings confirm that the apical membrane in the caecum and proximal colon of guinea pig is an effective barrier against a rapid diffusion of small lipid-soluble substances such as SCFAH. Besides cholesterol and membrane proteins, there are further factors that contribute to this barrier property.     

Prevalence of bacterial faecal pathogens in separated and unseparated stored pig slurry

AIMS: To examine the prevalence and diversity of bacterial faecal pathogens in unseparated slurry, separated solids and liquid fractions from a commercial pig farm. METHODS: A total of 43 stored slurry specimens originating from a fattening house over the period February-April 2002 were analysed, consisting of unseparated (n = 14) slurry, separated solids (n = 16) and separated liquid (n = 13). Specimens were examined for the presence of five bacterial pathogens including Salmonella spp., Shigella spp., Campylobacter spp., Escherichia coli O157 and Yersinia enterocolitica. Selective enrichment and plating methods were employed for detection of Salmonella spp. and Campylobacter spp. and conventional selective plating techniques for the remaining genera. Antibiogram profiles to 12 antibiotic agents were obtained for all Salmonella isolates obtained. RESULTS: Salmonella spp. were identified in all components of the slurry specimens, whereas Campylobacter spp. was only recovered from the unseparated and separated liquid fractions. In both cases, the separated liquid fraction had the highest prevalence of pathogens and the separated solid fraction had the lowest prevalence. None of the slurry specimens examined were positive for E. coli O157:H7, Shigella spp. or Y. enterocolitica. Twenty-nine isolates of Salmonella were recovered from the slurry specimens, comprising seven serovars, of which Salmonella manhattan was the most prevalent, accounting for over half [15 of 29 (51.7%)] of all Salmonella isolates. Salmonella anatum, Salm. derby, Salm. give, Salm. heidelberg, Salm. simi and Salm. stanley serovars were also recovered. All Salmonella isolates were sensitive to ampicillin, augmentin (amoxicillin/clavulanic acid), chloramphenicol, ciprofloxacin, gentamicin, kanamycin and trimethoprim, but has variable resistance to tetracycline (100%), sulphonamides (84.6%), furazolidone (38.5%), nalidixic acid (15.4%) and streptomycin (15.4%). The majority (57.7%) of isolates displayed antibiotic resistance to at least two antibiotic agents, followed by 34.6% of isolates being resistant to three agents and the remainder (7.7%) being resistant to four antibiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated a marked reduction in the prevalence of Campylobacter and Salmonella in the solids component of separated pig slurry. The adoption of control processes such as aeration of slurry prior to its spread onto agricultural land and newer approaches to pathogen reduction should be investigated, to reduce the transmission of pathogens from pig slurry to the environment.     

Potato powders prepared by successive cooking-process depending on resistant starch content affect the intestinal fermentation in rats

The effects of resistant starch (RS) in dry potato powders prepared by various processes on intestinal fermentation in rats were assessed. Rats were fed raw potato powder (RP), blanched potato powder (BP), steamed potato powder (SP), or drum-dried potato powder (DP) for 4 weeks. The cecal RS content was significantly higher in the RP group than in the control diet (CN) group and other dry potato powder groups. Cecum pH was significantly lower in the RP group compared to the CN group, and was also significantly lower than that in the SP, BP, and DP groups. Lactic acid bacteria levels in the RP group were significantly higher than those in the CN group, and levels in the SP group also increased relative to the control group. Lactobacillus levels in the RP group were higher than in the CN and other dry potato powder groups. Cecal short-chain fatty acid (SCFA) concentrations in the RP group followed by the SP group exhibited significantly higher levels relative to the control levels. Dry potato powders containing RS produced during the cooking process may represent a useful food material that increases intestinal concentrations of SCFA and enhances the growth of certain lactic acid bacteria.     

Anti-plasticization of cassava starch by complexing fatty acids

The effect of adding 1–8% amylose complexing fatty acids (CFA), such as linoleic and oleic acids, on the glass transition temperature (T_g) of cassava starch (CS) with moisture content varying from 5 to 35% (dry basis) was studied. The main relaxation temperature (T_α), associated with the glass transition temperature of the samples (T_g), was determined by dynamic-mechanical-thermal analysis. The plasticizing behavior of water in the blends was evidenced by a decrease of T_α values with moisture content. The effect of CFA on CS was found to be a function of moisture content. At low moisture (<11%) it caused an anti-plasticization effect, while at higher moisture contents it produced plasticization. The anti-plasticizing effect of CFA on CS was attributed to amylose–lipid complex formation.     

小麦籽粒抗性淀粉含量的分析

选用3个抗性淀粉含量较高的小麦品种和3个抗性淀粉含量较低的小麦品种按Griffing双列杂交设计配置成15个杂交组合, 以亲本及F₁为材料进行了小麦籽粒抗性淀粉含量的遗传规律分析, 旨在为高抗性淀粉含量且综合性状优良的新型保健小麦新品种(系)的选育提供理论依据。结果表明, 在6个小麦品种中, 安农90202和D68-20抗性淀粉含量的一般配合力较好, 能显著地提高杂种后代籽粒抗性淀粉含量。安农90202×04单28和06-5×D68-20组合的特殊配合力最好, 两者特殊配合力效应值显著地高于其他组合。小麦抗性淀粉含量的遗传符合加性-显性模型, 显性程度为超显性。控制抗性淀粉含量的增效等位基因表现为隐性, 且亲本中抗性淀粉含量的增减效等位基因的分布不平衡, 高抗性淀粉含量的亲本中隐性基因数量多于显性基因数量。实验中安农90202和04单28控制抗性淀粉含量的隐性基因较多,而宁春18和新春5号含有的显性基因数量较多。同时研究发现小麦抗性淀粉含量的狭义遗传力中等, 为36.49%。     

不同碳水化合物对孤独症患儿肠道菌群 体外发酵产生短链脂肪酸的影响

目的探究孤独症谱系障碍(autism spectrum disorder,ASD)患儿与健康儿童的肠道菌群利用不同碳水化合物产生短链脂肪酸(short-chain fatty acids,SCFAs)的差异。方法前瞻性病例对照研究。选择2016年12月至2017年10月解放军总医院儿科门诊的15例ASD患儿为研究对象,并匹配15名健康儿童为对照组;收集研究对象粪便,稀释成10%悬浊液,然后分别接种到以乳果糖(LAU)、乳糖(LAT)、棉籽糖(RAF)、低聚半乳糖(GOS)、低聚异麦芽糖(IMO)和低聚甘露糖(MOS)为单碳源的肠道微生态小型模拟发酵系统中批量发酵24 h,检测SCFAs浓度、底物降解率和产气气压。结果 ASD组患儿粪便SCFAs浓度与对照组差异无统计学意义,在YCFA(无碳源对照培养基,蛋白发酵为主)培养基发酵后总SCFAs、乙酸和丙酸均低于对照组,差异有统计学意义(Z=-2.509、-2.509、-3.007,P=0.011、0.011、0.002);在添加LAT、RAF、GOS、IMO和MOS的培养基发酵后总SCFAs浓度显著高于对照组,差异有统计学意义(Z=2.385、2.344、2.675、2.344、2.302,P=0.016、0.019、0.007、0.019、0.021),而含LAU的培养基发酵后,两组研究对象总SCFAs浓度比较差异无统计学意义。结论 ASD患儿肠道菌群利用蛋白发酵产SCFAs能力显著低于健康对照,利用碳水化合物产SCFAs能力显著高于健康对照。6种发酵底物中,乳果糖是最适合ASD患儿的碳水化合物,有改善ASD患儿肠道菌群产SCFAs的潜力。     

Predicting feed digestibility from NIRS analysis of pig faeces

Digestibility is a key parameter in the evaluation of feeds; however, the measurements on animals require heavy experimental trials, which are hardly feasible when large numbers of determinations are required – for example, in genetic studies. This experiment aimed at investigating the possibility to predict digestibility from NIRS spectra measured on faeces. A total of 196 samples were available from a digestibility experiment investigating the effects of age and genetic background of Large White pigs fed the same diet, rich in fibre (NDF=21.4% DM). Digestibility of dry matter (dDM), organic matter (dOM), nitrogen content (dN), energy (dE) and apparent digestible energy content (ADE) were calculated, as well as total N content of faeces (N). The faeces samples were submitted to reflectance NIRS analysis after freeze-drying and grinding. Calibration errors and validation errors were, respectively, 0.08 and 0.13% DM for total N in faeces, 0.97% and 1.08% for dDM, 0.79% and 1.04% for dOM, 1.04% and 1.47% for dN, 0.87% and 1.12% for dE and 167 and 213 kJ/kg DM for ADE. These results indicate that NIRS can account for digestibility differences due to animal factors, with an acceptable accuracy. NIRS appears to be a promising tool for large-scale evaluations of digestibility. It could also be used for the study of digestibility of different feeds, after appropriate calibration based on a wide range of feed types.     

The potential of resistant starch as a prebiotic

Resistant starch is defined as the total amount of starch and the products of starch degradation that resists digestion in the small intestine. Starches that were able to resist the digestion will arrive at the colon where they will be fermented by the gut microbiota, producing a variety of products which include short chain fatty acids that can provide a range of physiological benefits. There are several factors that could affect the resistant starch content of a carbohydrate which includes the starch granule morphology, the amylose–amylopectin ratio and its association with other food component. One of the current interests on resistant starch is their potential to be used as a prebiotic, which is a non-digestible food ingredient that benefits the host by stimulating the growth or activity of one or a limited number of beneficial bacteria in the colon. A resistant starch must fulfill three criterions to be classified as a prebiotic; resistance to the upper gastrointestinal environment, fermentation by the intestinal microbiota and selective stimulation of the growth and/or activity of the beneficial bacteria. The market of prebiotic is expected to reach USD 198 million in 2014 led by the export of oligosaccharides. Realizing this, novel carbohydrates such as resistant starch from various starch sources can contribute to the advancement of the prebiotic industry.     

In vitro stimulation by retinol of porcine pancreatic esterase activity toward esters of short-chain fatty acids.

1. Retinol exerted a remarkable stimulating effect (approx. 260% increase), essentially similar to that (300%) of phytol, on the so-called esterase activity displayed by crude pancreatic lipase [EC 3.1.1.3] toward true solutions of esters, but none of the typical lipase activity toward emulsions of water-insoluble esters. 2. Comparison of the stimulatory effects of retinol derivatives on the esterase activity revealed that retinyl acetate was the most active, being sustantially similar in effect to retinol; retinal was fairly active, while retinoic acid, retinyl palmitate, and beta-ionone were far less active. 3. With various isoprenoid compounds, the efficiency of stimulation increased with the carbon chain length, attaining a maximum at 15 to 20 carbon atoms. Above this chain length the efficiency decreased rapidly. 4. Comparison of the effects of retinol and phytol on the esterase activity of various other lipolytic enzymes indicated that this kind of activator may be relatively specific to porcine pancreatic esterase activity.     

Identification of a free fatty acid receptor,FFA2R,expressed on leukocytes and activated by short-chain fatty acids

Short-chain fatty acids (SCFAs) have long been known to exert cellular effects on blood leukocytes. Acetate, propionate, and butyrate represent the most capable SCFA, inducing calcium mobilization which subsequently regulates leukocyte function in the immune system. We have cloned the previously described putative orphan G-protein coupled receptor, GPR43, and have functionally identified SCFA as the activating ligands. Acetate and propionate were found to be the two most potent ligands, although butyrate, formate, and valerate (in this order of potency) also were able to induce receptor activation. Both the human and mouse receptor homologues were found to share the same pattern of ligand activation. This finding, together with a high degree of amino acid sequence similarity between the mouse and human homologues, indicates an evolutionary conserved function. Upon ligand stimulation, the receptor mobilized intracellular calcium in both a recombinant system as well as in human granulocytes. We found the human gene to be predominantly expressed in peripheral blood leukocytes and, to a lesser extent, in spleen. We suggest the designation FFA(2)R to this second receptor activated by free fatty acids. The first-described FFAR, now named FFA(1)R, is activated by medium- to long-chain free fatty acids.     

Suppressive effect of short-chain fatty acids on production of proinflammatory mediators by neutrophils

Short chain fatty acids (SCFAs) are fermentation products of anaerobic bacteria. More than just being an important energy source for intestinal epithelial cells, these compounds are modulators of leukocyte function and potential targets for the development of new drugs. The aim of this study was to evaluate the effects of SCFAs (acetate, propionate and butyrate) on production of nitric oxide (NO) and proinflammatory cytokines [tumor necrosis factor α (TNF-α) and cytokine-induced neutrophil chemoattractant-2 (CINC-2αβ)] by rat neutrophils. The involvement of nuclear factor κB (NF-κB) and histone deacetylase (HDAC) was examined. The effect of butyrate was also investigated in vivo after oral administration of tributyrin (a pro-drug of butyrate). Propionate and butyrate diminished TNF-α, CINC-2αβ and NO production by LPS-stimulated neutrophils. We also observed that these fatty acids inhibit HDAC activity and NF-κB activation, which might be involved in the attenuation of the LPS response. Products of cyclooxygenase and 5-lipoxygenase are not involved in the effects of SCFAs as indicated by the results obtained with the inhibitors of these enzymes. The recruitment of neutrophils to the peritonium after intraperitoneal administration of a glycogen solution (1%) and the ex vivo production of cytokines and NO by neutrophils were attenuated in rats that previously received tributyrin. These results argue that this triglyceride may be effective in the treatment of inflammatory conditions.     

Fermentation of prebiotics by human colonic microbiota in vitro and short-chain fatty acids production: a critical review

Prebiotics are known for their health benefits to man, including reducing cardiovascular disease and improving gut health. This review takes a critical assessment of the impact of dietary fibres and prebiotics on the gastrointestinal microbiota in vitro. The roles of colonic organisms, slow fermentation of prebiotics, production of high butyric and propionic acids and positive modulation of the host health were taken into cognizance. Also, the short-chain fatty acids (SCFAs) molecular signalling mechanisms associated with their prebiotic substrate structural conformations and the phenotypic responses related to the gut microbes composition were discussed. Furthermore, common dietary fibres such as resistant starch, pectin, hemicelluloses, β-glucan and fructan in context of their prebiotic potentials for human health were also explained. Finally, the in vitro human colonic fermentation depends on prebiotic type and its physicochemical characteristics, which will then affect the rate of fermentation, selectivity of micro-organisms to multiply, and SCFAs concentrations and compositions.     

小豆抗性淀粉含量及蒸煮后硬度分析

本文利用国内140个红小豆种质资源,探究其抗性淀粉含量与蒸煮后硬度的地域分布特征,分析蒸煮后硬度与营养指标的相关性,同时筛选抗性淀粉含量高与蒸煮后硬度低的种质资源,结果表明：140份红小豆抗性淀粉的平均含量为14.25 %,蒸煮后硬度的平均值为150.72 g。吉林地区红小豆抗性淀粉含量最高,为15.71 %;内蒙古地区红小豆蒸煮后硬度最低,为96.42 g。抗性淀粉与总淀粉之间呈极显著负相关;蒸煮后硬度与抗性淀粉之间呈显著正相关,但与总淀粉及蛋白质之间无显著相关。筛选出12份抗性淀粉含量>17.83 %的优异红小豆种质资源,可用于糖尿病人专用品种的选育及产品开发;9份蒸煮后硬度<76.48 g的优异红小豆种质资源可用于豆饭、豆粥产品的开发。     

Influence of short-chain fatty acids on the production of spiramycin by Streptomyces ambofaciens

Summary The addition of short-chain fatty acids stimulates the production of spiramycin by Streptomyces ambofaciens cultivated on dextrins and ammonium chloride. The fatty acids were activated by two enzymatic systems. The first system (acyl-CoA synthetases) was present only during the exponential phase. The second system (acylkinases coupled with acylphosphotransferases) was synthesized during the growth phase and during the stationary phase, in which spiramycin production started. Short-chain fatty acids induced the synthesis of acylkinases and acylphosphotransferases. Added at the beginning of cultures, they increased the specific activity of these enzymes during the exponential growth phase. Added at the early stationary phase, the specific activity of these enzymes and of the spiramycin production increased. Excess ammonium in the culture considerably lowered the specific activity of acylkinases synthesized in the stationary phase, when spiramycin productiin started. This ammonium effect can be reduced by the addition of short-chain fatty acids.Offprint requests to: A. Lebrihi     

NMR study of 13CO2 incorporation into short-chain fatty acids by pig large-intestinal flora

The nuclear magnetic resonance technique was used to study carbon dioxide reduction by the pig large-intestinal flora. Washed bacterial cell suspensions were incubated for 6 and 15 h under 13CO2 and H2 as the gas phase and with a buffer containing NaH13CO3 and cellobiose and amino acids (casein hydrolysate) as substrates. Methane was produced in all incubation media. Significant amounts of single- as well as multiple-labelled acetate and butyrate were formed, demonstrating synthesis of acetate from H2 + CO2. Propionate was labelled mainly on the carboxyl group, which was attributed to an enzymatic exchange of the carboxyl group of propionate with 13CO2. These results indicate that the reduction of CO2 to acetate may be an important pathway for microbial production of acetate in the pig large intestine even in the presence of methanogenesis.     

Development of formulae for estimating amylose content and resistant starch content based on the pasting properties measured by RVA of Japonica polished rice and starch

We searched for the easy and simple method to measure the novel indicators which reflect not only AAC, but also (RS) based on pasting properties using RVA. Novel indexes such as SB/Con and Max/Fin (Maximum viscosity/Minimum viscosity) ratios had a very high correlation with proportion of intermediate and long chains of amylopectin; Fb₁₊₂₊₃ (DP ≧ 13). In Japonica polished rice, estimation formulae for AAC and RS content were developed using novel indexes based on pasting properties by RVA, and these equations showed determination coefficients of 0.89 and 0.80 for calibration and 0.71 and 0.75 for validation test. We developed the estimation formulae for AAC and RS content for Japonica starch samples. These equations showed determination coefficients of 0.86 and 1.00 for calibration and 0.76 and 0.83 for validation test, which showed that these equations can be applied to the unknown rice samples.     

The regulatory effects of resistant starch on glycaemic response in obese dogs

The purpose of this study was to examine the inhibitory effects of resistant starch on postprandial glycaemic response in obese dogs. The changes in blood glucose concentrations and glycaemic index (GI) were chronologically determined after the administration of resistant and normal starches by nasal feeding. Resistant starch contained indigestible dextrin (IDD) and β-cyclic dextrin (β-CD). Soluble starch (SS) served as a control starch. Glucose concentrations reached their maximum 15 min after the administration of SS solutions, and decreased gradually during the experimental period. In contrast, after the administration of IDD solutions, increased glucose concentrations rapidly decreased to the initial values. After the administration of β-CD solutions, glucose concentrations remained unchanged during this study. GI levels remained constant in the following order: β-CD < IDD < SS. GI levels of dogs receiving IDD and β-CD solutions were significantly lower as compared with those animals receiving SS solutions. In this study, nasal tube feeding was an effective method for evaluating glycaemic responses to various starches accurately. The present data revealed that resistant starches were useful materials in controlling nutritionally glucose concentrations in obese dogs. These results raise the possibility that resistant starches are valuable for dietetic treatment of diabetes and obesity in dogs.