Clinical uses of probiotics for stabilizing the gut mucosal barrier: successful strains and future challenges

Probiotic bacteria are used to treat disturbed intestinal microflora and increased gut permeability which are characteristic to many intestinal disorders. Examples include children with acute rotavirus diarrhoea, subjects with food allergy, subjects with colonic disorders and patients undergoing pelvic radiotherapy and sometimes changes associated with colon cancer development. In all such disease states altered intestinal microflora, impaired gut barrier and different types of intestinal inflammation are present. Successful probiotic bacteria are able to survive gastric conditions and colonize the intestine, at least temporarily, by adhering to the intestinal epithelium. Such probiotic microorganisms appear to be promising candidates for the treatment of clinical conditions with abnormal gut microflora and altered gut mucosal barrier functions. They are also promising ingredients to future functional foods and clinical foods for specific disease states provided that basic requirements for strains and clinical studies are carefully followed.     

Bacteroides thetaiotaomicron: a dynamic, niche-adapted human symbiont

The coevolution of humans with their intestinal microflora has resulted in cooperative relationships that have shaped the biology and the genomes of these symbiotic partners. Bacteroides thetaiotaomicron is one such bacterial symbiont that is a dominant member of the intestinal microbiota of humans and other mammals. The recent report of the genome sequence of B. thetaiotaomicron is the first reported for an abundant Gram-negative organism of the human colonic microbiota and, as such, provides the first glimpse on a genomic scale of the genetic arsenal used by a Gram-negative symbiont to dominate in this ecosystem. The genome has revealed large expansions of many paralogous groups of genes that encode products essential to the organism's ability to successfully compete in this environment. Most noteable is the organism's abundant machinery for utilizing a large variety of complex polysaccharides as a source of carbon and energy. The proteome also reveals the organism's extensive ability to adapt and regulate expression of its genes in response to the changing ecosystem. These factors, as well as others highlighted below, suggest an incredibly flexible and adaptable organism that is exquisitely equipped to dominate in its challenging and competitive niche.     

Gastric acid reduction leads to an alteration in lower intestinal microflora

To clarify the alterations in lower intestinal microflora induced by gastric acid reduction, the dynamics of 12 major genera or groups of bacteria comprising the microflora in feces and colonic contents were examined by quantitative real-time PCR in proton pump inhibitor-treated rats and in asymptomatic human subjects with hypochlorhydria. In both rat and human experiments, most genera or groups of intestinal microflora (facultative and obligate anaerobes) proliferated by gastric acid reduction, and marked and significant increases in the Lactobacilli group and Veillonella, oropharyngeal bacteria, were observed. In rats, potent gastric acid inhibition led to a marked and significant increase of intestinal bacteria, including the Bacteroidesfragilis group, while Bifidobacterium, a beneficial bacterial species, remained at a constant level. These results strongly indicate that the gastric acid barrier not only controls the colonization and growth of oropharyngeal bacteria, but also regulates the population and composition of lower intestinal microflora.     

益生元对抗生素引起的肠道菌群失调的作用

目的 评价益生元菊糖和大蒜多糖在治疗肠道菌群失调中的作用.方法 72只Balb/c小鼠随机分为6组,随机取出12只作为正常对照组,其余灌胃盐酸林可霉素和头孢曲松钠混合抗生素溶液3d,建立小鼠肠道失调模型后,以生理盐水灌胃作为自然恢复组,其余4组分别灌胃菊糖、低剂量大蒜多糖、高剂量大蒜多糖以及丽珠肠乐5d.受试小鼠无菌取粪,观察各组肠道菌群的数量变化、测量血清IgG,粪便内毒素含量、有机酸含量变化.结果 菊糖和大蒜多糖高剂量灌胃能加速恢复抗生素导致的小鼠肠道菌群失调,这两种益生元对内毒素的清理、免疫力的恢复也有积极的效果,效果均优于丽珠肠乐和自动恢复.结论 益生元有加快肠道菌群恢复和降低抗生素的破坏效果,有益于小鼠肠道健康.     

Non-starch polysaccharides extracted from seaweed can modulate intestinal absorption of glucose and insulin response in the pig

We have investigated the possible effects of algal polysaccharides on postprandial blood glucose and insulin responses in an animal model, the pig. Three seaweed fibres of different viscosities, extracted from Palmaria palmata (PP), Eucheuma cottonii (EC), or Laminaria digitata (LD), were compared to purified cellulose (CEL). Blood glucose and plasma insulin levels were monitored and intestinal absorption quantified for 8 h following a high carbohydrate test-meal supplemented with 5% fibre. Digestive contents were also sampled, 5 h postprandial. As compared to CEL, PP had no effect on glucose and insulin responses. The latter decreased with EC, but glucose absorption balance was not modified. LD addition resulted in a dramatically reduced glucose absorption balance, accompanied by a higher amount of starch left in the small intestine. Among polysaccharides tested, only the highly viscous alginates could affect intestinal absorption of glucose and insulin response.     

海藻粉和抗菌肽对肉鸡生长性能、免疫器官指数和肠道菌群的影响

为探讨海藻粉和抗菌肽不同添加水平对肉鸡生长性能、免疫器官指数和肠道菌群的影响。选取1日龄体重相近的AA肉鸡504羽,随机分为7个处理,每处理6个重复,每重复12羽。对照组饲喂基础日粮,试验组在基础日粮中添加不同水平(1.0%、3.0%、5.0%)的海藻粉和抗菌肽(300 mg/kg、600 mg/kg)。试验分前期(1~21 d)和后期(22~42 d)共42 d。结果表明:(1)添加不同水平的海藻粉对前期、后期和全期肉鸡的料重比有显著影响(P0.05);对21日龄肉鸡的脾脏指数和胸腺指数及对42日龄肉鸡的法氏囊指数差异显著(P0.05);降低了21日龄和42日龄肉鸡盲肠大肠杆菌的数量(P0.05),增加了盲肠乳酸菌的数量(P0.05)。(2)添加不同水平的抗菌肽对后期和全期肉鸡的料重比有显著差异(P0.05);对21日龄肉鸡的脾脏指数和胸腺指数及对42日龄肉鸡的法氏囊指数差异显著(P0.05);降低了21日龄和42日龄肉鸡盲肠大肠杆菌的数量(P0.05),增加了盲肠乳酸菌的数量(P0.05)。(3)海藻粉和抗菌肽交互作用对前期、后期和全期肉鸡的料重比有明显影响(P0.05);对21日龄肉鸡的脾脏指数和胸腺指数及对42日龄肉鸡的法氏囊指数差异显著(P0.05);降低了21日龄和42日龄肉鸡盲肠大肠杆菌的数量(P0.05),增加了盲肠乳酸菌的数量(P0.05)。本研究得出以下结论,海藻粉和抗菌肽能促进肉鸡生长,提高肉鸡免疫器官指数和改善肠道菌群,且二者间存在协同效应,添加5%的海藻粉和300 mg/kg抗菌肽综合效果最佳。     

Antiviral polysaccharides isolated from Hong Kong brown seaweed <Emphasis Type="Italic">Hydroclathrus clathratus</Emphasis>

Two relatively pure polysaccharides H3-a1 and H3-b1 had been isolated from the brown seaweed Hydroclathrus clathratus. They were characterized by HPLC, ultraviolet scanning, gas chromatography, infrared spectroscopy and elemental analysis, and shown to be two different sulfated polysaccharides with different monosaccharide content, but both with high relative molecular mass. They contained some proteins and uronic acid respectively. The sulfate content and bioactivity of these polysaccharides varied during purification. The fractions derived from the hot water extract also exhibited low anticoagulant effect. This is the first time that the antiherpetic and anticoagulant activities were evaluated for the polysaccharides from the Hong Kong brown seaweed Hydroclathrus clathratus.     

Berberine promotes recovery of colitis and inhibits inflammatory responses in colonic macrophages and epithelial cells in DSS-treated mice

Inflammatory bowel disease (IBD) results from dysregulation of intestinal mucosal immune responses to microflora in genetically susceptible hosts. A major challenge for IBD research is to develop new strategies for treating this disease. Berberine, an alkaloid derived from plants, is an alternative medicine for treating bacterial diarrhea and intestinal parasite infections. Recent studies suggest that berberine exerts several other beneficial effects, including inducing anti-inflammatory responses. This study determined the effect of berberine on treating dextran sulfate sodium (DSS)-induced intestinal injury and colitis in mice. Berberine was administered through gavage to mice with established DSS-induced intestinal injury and colitis. Clinical parameters, intestinal integrity, proinflammatory cytokine production, and signaling pathways in colonic macrophages and epithelial cells were determined. Berberine ameliorated DSS-induced body weight loss, myeloperoxidase activity, shortening of the colon, injury, and inflammation scores. DSS-upregulated proinflammatory cytokine levels in the colon, including TNF, IFN-γ, KC, and IL-17 were reduced by berberine. Berberine decreased DSS-induced disruption of barrier function and apoptosis in the colon epithelium. Furthermore, berberine inhibited proinflammatory cytokine production in colonic macrophages and epithelial cells in DSS-treated mice and promoted apoptosis of colonic macrophages. Activation of signaling pathways involved in stimulation of proinflammatory cytokine production, including MAPK and NF-κB, in colonic macrophages and epithelial cells from DSS-treated mice was decreased by berberine. In summary, berberine promotes recovery of DSS-induced colitis and exerts inhibitory effects on proinflammatory responses in colonic macrophages and epithelial cells. Thus berberine may represent a new therapeutic approach for treating gastrointestinal inflammatory disorders.     

Antiviral polysaccharides isolated from Hong Kong brown seaweed Hydroclathrus clathratus

Two relatively pure polysaccharides H3-a1 and H3-b1 had been isolated from the brown seaweed Hy-droclathrus clathratus. They were characterized by HPLC, ultraviolet scanning, gas chromatography, infrared spectroscopy and elemental analysis, and shown to be two different sulfated polysaccharides with different monosaccharide content, but both with high relative molecular mass. They contained some proteins and uronic acid respectively. The sulfate content and bioactivity of these polysaccha-rides varied during purification. The fractions derived from the hot water extract also exhibited low anticoagulant effect. This is the first time that the antiherpetic and anticoagulant activities were evaluated for the polysaccharides from the Hong Kong brown seaweed Hydroclathrus clathratus.     

肠道菌群对糖脂代谢影响的研究进展

目前越来越多的研究成果表明被誉为人类"第二基因组"的肠道菌群能够影响糖脂代谢,进而调节相应疾病。本文就肠道菌群的分布、种类、影响因素进行大体介绍,主要对肠道菌群与糖脂代谢疾病联系和肠道菌群调节糖脂代谢过程中可能存在的作用机理进行综述,希望能够对糖脂代谢疾病的临床诊治带来裨益,为在此领域上的研究者提供参考价值。     

Variation of mucin distribution in the rat intestine, caecum and colon: effect of the bacterial flora.

The influence of the intestinal microflora on mucin types was studied in the small intestine, caecum and colon of conventional (CV) rats as compared to germ-free (GF) rats. A colorimetric method was used on purified water-soluble mucin extracted from mucosal scrapings and contents. Variations occurred between the three anatomical sites both in the mucosas and intestinal contents of GF rats. In CV rats, the presence of the bacterial flora led to different effects depending on the intestinal site: in the small intestinal mucosa, neutral and sulphomucins values were higher whereas sialomucin was much lower. Conversely, sialomucin was higher in the caecal and colonic mucosas and contents whereas sulphated mucins were decreased significantly in caecal contents and caecal and colonic mucosas. These variations in the contents may reflect the bacterial mucolytic activity and the effect of bacterial metabolites on the mucosa.     

The effect of seaweed composite flour on the textural properties of dough and bread

Seaweeds as food and seaweed-derived food flavors, colors, and nutrients are attracting considerable commercial attention. In the baking industries, hydrocolloids are of increasing importance as bread making improvers, where their use aims to improve dough handling properties, increase the quality of fresh bread, and extend the shelf life of stored bread. Seaweeds contain a significant amount of soluble polysaccharides and have the potential function as a source of dietary fiber. In this study, red seaweed (Kappaphycus alvarezii) powder was incorporated (2–8 %) with wheat flour and used to produce bread. The effect of seaweed composite flour on dough rheological properties and the quality of bread was investigated using various techniques. Farinograph tests were applied to determine the effect of seaweed powder on the rheological properties of wheat flour dough, while texture profile analysis (TPA) was used to measure the textural properties of dough as well as the final product. The results showed that the additions of seaweed powder (2–8 %) increased the water absorption of the dough. TPA results showed that the addition of seaweed powder decreased stickiness properties. Bread produced with seaweed composite flour showed higher values of firmness.     

发酵饲料对生长育肥猪结肠微生物发酵及菌群组成的影响

【目的】研究复合菌发酵饲料对生长育肥猪结肠发酵、结肠黏膜与结肠内容物菌群组成的影响。【方法】采用气相色谱法检测育肥猪结肠内容物中挥发性脂肪酸浓度;采用MiSeq高通量测序方法检测育肥猪结肠黏膜与内容物中细菌菌群组成。【结果】饲喂发酵饲料对结肠黏膜及内容物中菌群多样性无显著影响(P0.05);显著提高了猪结肠黏膜中魏斯菌属和柔嫩梭菌属的相对丰度(P0.05),提高了结肠内容物中魏斯菌属、Subdoligranulum菌属相对丰度(P0.05);饲喂发酵饲料对结肠内容物中pH、乳酸、乙酸、丙酸、异丁酸、戊酸、异戊酸和总挥发性脂肪酸浓度无显著影响(P0.05),但显著提高了结肠内容物中的丁酸水平(P0.05)。【结论】饲喂复合菌发酵饲料可在一定程度上影响育肥猪结肠中细菌菌群的组成,促进丁酸生成,对肠道健康具有改善作用。     

西藏灵菇奶对抗生素相关性腹泻小鼠肠道菌群的调整作用

目的研究用西藏灵菇对抗生素引起的肠道菌群失调及病理变化的调整作用.方法由林可霉素造成小鼠腹泻模型,并证实肠道菌群紊乱后,将其分成灵菇奶治疗组和自然恢复组,7 d后对每个组小鼠进行肠道菌群检测,同时取回肠末段组织标本用光镜及电镜观察黏膜结构的变化.结果抗生素模型组大多数小鼠肠道黏膜出现不同程度的损害,小鼠肠道的正常菌群及病理组织改变基本恢复正常.结论灵菇奶不仅对肠道菌群有调整作用,而且对肠黏膜的修复有一定作用.     

Isolation and characterization of an alginase from mucoid strains of Pseudomonas aeruginosa.

Strains of Pseudomonas aeruginosa which produce an alginate-like slime polysaccharide were shown to also synthesize an intracellular enzyme which can degrade these polysaccharides and the seaweed alginic acids. The enzyme acts as an eliminase introducing delta 4,5 unsaturation into the uronic acid moiety. It appears to be a polymannuronide lyase which degrades the polysaccharides, depending on their uronic acid composition, to a series of oligosaccharides, the smallest of which is a disaccharide. L-Guluronic acid linkages are not split. The Pseudomonas alginase resembles other bacterial alginases and enzymes from molluscs but differs in some important properties, such as extent of degradation and linkage preference. Nonmucoid forms of the organism produce detectable but much lower amounts of enzyme.     

Morphological and physiological characteristics of bacteria inhabiting the intestinal mucosa of pike (<Emphasis Type="Italic">Esox lucius</Emphasis> L.)

Predominance of gram-negative bacteria belong ing to the class Gammaproteobacteria on the intestinal mucosa of pike was determined. The morphophysiological features of the isolated microorganisms suggest that they belong to symbiotic intestinal microflora. The morphological characteristics of the intestinal symbiotic microflora of pike include the formation of the capsule, pseudovacuoles, and spheroplastic forms of cells. The bacteria that were found can produce hydrolytic enzymes and possess persistence properties. The morphophysiological features that were found contribute to the adaptation of bacteria to environmental conditions and indicate that the intestinal microflora offish is autochthonous.     

Effect of Processing on the Microflora of Norwegian Seaweed Meal, with Observations on Sporendonema minutum (Høye) Frank and Hess

Meal from the brown seaweed Ascophyllum nodosum (L.) Le Jol. is mainly used as an animal feed supplement. Since moist weed often develops a marked mold growth and since little was known about the microflora of seaweed meal, a cultural procedure was developed to enumerate the populations of bacteria, yeasts, and molds of seaweed meals manufactured by different drying processes. The microflora could be supported by a variety of media varying in levels of nutrition and in the source and concentration of salts. Fresh weed contained less than 10(3) bacteria and less than 10(2) yeasts and molds per g (dry weight). The type and extent of microbial populations in seaweed meal appeared to be dependent upon the method of seaweed drying. Rotary drum-drying at temperatures decreasing from 800 to 80 C maintained or reduced the microbial populations to 10(3) organisms per g (dry weight). Although meals with high nutritional quality can be obtained with warm air- or rock-dried weed, these conditions can also permit bacterial and mold development. Extended rock-drying in variable weather conditions and prolonged storage of moist weed, both of which decrease the nutritional quality, also lead to high bacterial numbers and to a marked development of the halophilic brown mold Sporendonema minutum which attained populations of 10(8) viable spores per g of dried weed. A poultry diet containing 5% badly molded weed had no apparent toxic or growth-depressing effect when fed to chicks.     

The potential of bacteria isolated from ruminal contents of seaweed‐eating North Ronaldsay sheep to hydrolyse seaweed components and produce methane by anaerobic digestion in vitro

The production of methane biofuel from seaweeds is limited by the hydrolysis of polysaccharides. The rumen microbiota of seaweed‐eating North Ronaldsay sheep was studied for polysaccharidic bacterial isolates degrading brown‐seaweed polysaccharides. Only nine isolates out of 65 utilized > 90% of the polysaccharide they were isolated on. The nine isolates (eight Prevotella spp. and one Clostridium butyricum) utilized whole Laminaria hyperborea extract and a range of seaweed polysaccharides, including alginate (seven out of nine isolates), laminarin and carboxymethylcellulose (eight out of nine isolates); while two out of nine isolates additionally hydrolysed fucoidan to some extent. Crude enzyme extracts from three of the isolates studied further had diverse glycosidases and polysaccharidase activities; particularly against laminarin and alginate (two isolates were shown to have alginate lyase activity) and notably fucoidan and carageenan (one isolate). In serial culture rumen microbiota hydrolysed a range of seaweed polysaccharides (fucoidan to a notably lesser degree) and homogenates of L. hyperborea, mixed Fucus spp. and Ascophyllum nodosum to produce methane and acetate. The rumen microbiota and isolates represent potential adjunct organisms or enzymes which may improve hydrolysis of seaweed components and thus improve the efficiency of seaweed anaerobic digestion for methane biofuel production.     

Intestinal release and uptake of phenolic antioxidant diferulic acids.

Diferulic acids are potent antioxidants and are abundant structural components of plant cell walls, especially in cereal brans. As such, they are part of many human and animal diets and may contribute to the beneficial effect of cereal brans on health. However, these phenolics are ester-linked to cell wall polysaccharides and cannot be absorbed in this form. This study provides the first evidence that diferulic acids can be absorbed via the gastrointestinal tract. The 5-5-, 8-O-4-, and 8-5-diferulic acids were identified in the plasma of rats after oral dosing with a mixture of the three acids in oil. Our study also reveals that human and rat colonic microflora contain esterase activity able to release 5-5-, 8-O-4-, and 8-5-diferulic acids from model compounds and dietary cereal brans, hence providing a mechanism for release of dietary diferulates prior to absorption of the free acids. In addition, cell-free extracts from human and rat small intestine mucosa exhibited esterase activity towards diferulate esters. Hence, we have shown that esterified diferulates can be released from cereal brans by intestinal enzymes, and that free diferulic acids can be absorbed and enter the circulatory system. Our results suggest that the phenolic antioxidant diferulic acids are bioavailable.