Immunological Properties of Inulin-Type Fructans

Beneficial effects of inulin-type fructans are discussed in view of studies that applied the oligosaccharides in colon cancer, chronic inflammatory diseases, vaccination efficacy, and prevention of infection and allergy. In the present paper, we discuss their immunomodulating effects. It is suggested that immunomodulation is elicited through indirect and direct mechanisms. Indirect mechanisms encompass stimulation of growth and activity of lactic acid bacteria, but can also be caused by fermentation products of these bacteria, i.e., short chain fatty acids. Evidence for direct effects on the immune system generally remains to be confirmed. It is suggested that inulin-type fructans can be detected by gut dendritic cells (DCs), through receptor ligation of pathogen recognition receptors (PRRs) such as Toll-like receptors, nucleotide oligomerization domain containing proteins (NODs), C-type lectin receptors, and galectins, eventually inducing pro- and anti-inflammatory cytokines. DCs may also exert antigen presenting capacity toward effector cells, such as B cells, T cells, and natural killer cells locally, or in the spleen. Inulin-type fructans may also ligate PRRs expressed on gut epithelium, which could influence its barrier function. Inulin-type fructans are potent immunomodulating food components that hold many promises for prevention of disease. However, more studies into the mechanisms, dose-effect relations, and structure-function studies are required.     

微生态制剂调节便秘、腹泻人群肠道菌群结构与产短链脂肪酸关键菌属的相关性

为探究微生态制剂对健康、便秘和腹泻人群肠道菌群结构的调节能力。本研究以健康、便秘和腹泻人群 为对象,令其定时、定量摄入水苏糖（stachyose tetrahydrate,Sta）、益生菌纯粉（probiotics power,PP）、益生 菌剂（probotic preparations,PPrs）3 种微生态制剂,共6 周,采集新鲜粪便样本并提取DNA,利用Ion torrent PGM 二代测序技术进行16S rRNA V3区扩增子测序,并用气相色谱检测粪便中的短链脂肪酸（short-chain fatty acids, SCFAs）表达水平,最后联合多变量统计学分析方法对测序数据进行多样性分析。结果表明：绝大多数序列属 于硬壁菌门（Firmicutes）和拟杆菌门（Bacteroidetes）,约占总序列数的94.37%。随着微生态制剂的摄入,受 试人群肠道菌群的群落结构多样性明显增加,毛螺菌科（Lachnospiraceae）中的Blautia、Lachnospira以及瘤胃菌 科（Ruminococcaceae）中的Faecalibacterium、Oscillospir等与产SCFAs相关的菌属都明显地增长,其中Blautia和 Faecalibacterium与SCFAs含量呈显著正相关。SCFAs的含量以及肠道中相应菌群的增长与减少都与微生态制剂的成 分相关,在3 组受试人群中,服用Sta组,丙酸含量显著增加,乙酸与丁酸含量也在2 周左右有所增加,并伴随产 SCFAs的菌属快速且大量增长;PP组肠道中只有乙酸含量有所增加,丙酸和丁酸含量呈现降低的趋势,而产SCFAs 的菌属增长较明显;服用PPrs组,乙酸和丁酸含量明显增加,且便秘和腹泻人群在停止服用后,其SCFAs的含量 接近于健康人群,常见的Bifidobacterium、Lactobacillus、Parabacteroides等外源性益生菌均明显增长,可能性致病 菌相对丰度降低,表明服用PPrs对肠道菌群结构的调节作用以及影响更大。此外,根据肠型的分析,Bacteroides和 Prevotella在饮食的共同驱动下会调整并改变肠型,而仅通过所选择的微生态制剂的作用,在驱动肠型改变方面不 显著。综上所述,肠道疾病状态的人群服用微生态制剂后,肠道菌群结构向正常人群的状态调整,菌群多样性和 SCFAs表达水平提高,表现出持续抑制肠道有害细菌生长,促进有益菌的增殖,以此来维持肠道菌群结构的稳态。 经扩增子测序分析获得初步结论为：微生态制剂有改变腹泻、便秘人群肠道菌群整体结构的功效,并且PPrs要比单 一的Sta或PP调整肠道菌群的能力更突出。     

酸枣仁水提物对焦虑大鼠症状的改善及其对肠道菌群的影响

选用慢性温和不可预知应激大鼠模型,探究酸枣仁水提物缓解焦虑症状的潜在作用机制。以行为学(糖水偏好率、旷场得分、暗处所处时间)及肠道菌群为参考指标,研究酸枣仁水提物对焦虑模型大鼠的行为、肠道菌群多样性、结构及组成的影响。结果显示,与模型组(CUMS)组相比,酸枣仁(SZS)组显著改善了慢性压力诱导的焦虑行为(糖水消偏好率及旷场得分显著增加,在暗箱中所处的时间显著减少);SZS组肠道菌群的alpha多样性显著升高(p0.05),且菌群结构显著偏离CUMS组;进一步探究发现,相比于CUMS组,SZS组中Lactobacillus(CUMS:29.42%;SZS:8.08%;p=0.003)和Ruminococcaceae_UCG-014(CUMS:3.30%;SZS:2.14%;p=0.04)的丰度显著降低,而Bacteroides(CUMS:2.23%;SZS:5.15%;p=0.001)、norank_o_Mollicutes_RF9(CUMS:1.01%;SZS:2.43%;p=0.019)、Ruminiclostridium_9(CUMS:0.42%;SZS:0.62%;p=0.034)、Parabacteroides(CUMS:0.35%;SZS:0.60%;p=0.011)、Butyricimonas(CUMS:0.01%;SZS:0.03%;p=0.018)及Acetatifactor(CUMS:0.01%;SZS:0.03%;p=0.001)的丰度显著升高。结果表明,酸枣仁可改善CUMS大鼠焦虑相关症状,且影响了CUMS大鼠的肠道菌群结构,为明确其潜在作用机制提供了理论依据。     

Expression of Main Toll-Like Receptors in Patients with Different Types of Colorectal Polyps and Their Relationship with Gut Microbiota

Abnormal activation of Toll-like receptor (TLRs) signaling can result in colon cancer development. The aim of this study was to investigate the expression of important TLRs in different histological types of colorectal polyps and evaluate their relationship with intestinal microbiota. The expression levels of TLR2, 3, 4, and 5 were analyzed in intestinal biopsy specimens of 21 hyperplastic polyp (HP), 16 sessile serrated adenoma (SSA), 29 tubular adenoma (TA), 21 villous/tubulovillous (VP/TVP) cases, and 31 normal controls. In addition, selected gut bacteria including Streptococcus bovis, Enterococcus faecalis, Enterotoxigenic Bacteroides fragilis (ETBF), Fusobacterium nucleatum, Porphyromonas spp., Lactobacillus spp., Roseburia spp., and Bifidobacterium spp. were quantified in fecal samples using absolute qRT PCR, and, finally, the association between TLRs and these gut microbiota- was evaluated by Spearman’s correlation coefficient. Higher expression of TLR2 and TLR4 in VP/TVP and TA, and lower expression levels of TLR3 and TLR5 in all type of polyps were observed. The differences in TLR expression patterns was not only dependent on the histology, location, size, and dysplasia grade of polyps but also related to the intestinal microbiota patterns. TLR2 and TLR4 expression was directly associated with the F. nucleatum, E. faecalis, S. bovis, Porphyromonas, and inversely to Bifidobacterium, Lactobacillus, and Roseburia quantity. Furthermore, TLR3 and TLR5 expression was directly associated with Bifidobacterium, Roseburia, and Lactobacillus quantity. Our results suggest a possible critical role of TLRs during colorectal polyp progression. An abnormal regulation of TLRs in relation to gut microbial quantity may contribute to carcinogenesis.     

Diet-Induced Gut Barrier Dysfunction Is Exacerbated in Mice Lacking Cannabinoid 1 Receptors in the Intestinal Epithelium

The gut barrier provides protection from pathogens and its function is compromised in diet-induced obesity (DIO). The endocannabinoid system in the gut is dysregulated in DIO and participates in gut barrier function; however, whether its activity is protective or detrimental for gut barrier integrity is unclear. We used mice conditionally deficient in cannabinoid receptor subtype-1 (CB₁R) in the intestinal epithelium (intCB1−/−) to test the hypothesis that CB₁Rs in intestinal epithelial cells provide protection from diet-induced gut barrier dysfunction. Control and intCB1−/− mice were placed for eight weeks on a high-fat/sucrose Western-style diet (WD) or a low-fat/no-sucrose diet. Endocannabinoid levels and activity of their metabolic enzymes were measured in the large-intestinal epithelium (LI). Paracellular permeability was tested in vivo, and expression of genes for gut barrier components and inflammatory markers were analyzed. Mice fed WD had (i) reduced levels of endocannabinoids in the LI due to lower activity of their biosynthetic enzymes, and (ii) increased permeability that was exacerbated in intCB1−/− mice. Moreover, intCB1−/− mice fed WD had decreased expression of genes for tight junction proteins and increased expression of inflammatory markers in LI. These results suggest that CB₁Rs in the intestinal epithelium serve a protective role in gut barrier function in DIO.     

Insect Gut Bacteria Promoting the Growth of Tomato Plants (Solanum lycopersicum L.)

Simple SummaryA new sources of plant growth-promoting (PGP) bacteria are needed to increase the agricultural crops without increasing the usage of chemicals. The aim of this study was to test the hypothesis that insect-gut bacteria promote tomato plant growth. The insect gut bacteria were screened for the presence of PGP traits. The plants treated with bacterial consortium showed a significant increase in fruit yield, in both number of fruits (+41%) and weight of fruits (+44%). Our results showed that: (i) D. virgifera gut’s bacteria significantly promote the growth of tomato plants, and (ii) bacteria other than plant-related can be considered as PGP. It must be underlined that even though the insect gut bacteria were proven to stimulate the plant’s growth, their practical usage must be preceded by an examination of their influence on ecological and biological safety.AbstractWe investigated gut bacteria from three insect species for the presence of plant growth properties (PGP). Out of 146 bacterial strains obtained from 20 adult specimens of Scolytidae sp., 50 specimens of Oulema melanopus, and 150 specimens of Diabrotica virgifera, we selected 11 strains displaying the following: PGP, phosphate solubility, production of cellulase, siderophore, lipase, protease, and hydrogen cyanide. The strains were tested for growth promotion ability on tomato (Lycopersicon esculentum) plants. Each strain was tested individually, and all strains were tested together as a bacterial consortium. Tomato fruit yield was compared with the negative control. The plants treated with bacterial consortium showed a significant increase in fruit yield, in both number of fruits (+41%) and weight of fruits (+44%). The second highest yield was obtained for treatment with Serratia liquefaciens Dv032 strain, where the number and weight of yielded fruits increased by 35% and 30%, respectively. All selected 11 strains were obtained from Western Corn Rootworm (WCR), Diabrotica virgifera. The consortium comprised: Ewingella americana, Lactococcus garvieae, L. lactis, Pseudomonas putida, Serratia liquefaciens, and S. plymuthica. To our knowledge, this is the first successful application of D. virgifera gut bacteria for tomato plant growth stimulation that has been described.     

Early-Life Development of the Bifidobacterial Community in the Infant Gut

The establishment of the gut microbiota poses implications for short and long-term health. Bifidobacterium is an important taxon in early life, being one of the most abundant genera in the infant intestinal microbiota and carrying out key functions for maintaining host-homeostasis. Recent metagenomic studies have shown that different factors, such as gestational age, delivery mode, or feeding habits, affect the gut microbiota establishment at high phylogenetic levels. However, their impact on the specific bifidobacterial populations is not yet well understood. Here we studied the impact of these factors on the different Bifidobacterium species and subspecies at both the quantitative and qualitative levels. Fecal samples were taken from 85 neonates at 2, 10, 30, 90 days of life, and the relative proportions of the different bifidobacterial populations were assessed by 16S rRNA–23S rRNA internal transcribed spacer (ITS) region sequencing. Absolute levels of the main species were determined by q-PCR. Our results showed that the bifidobacterial population establishment is affected by gestational age, delivery mode, and infant feeding, as it is evidenced by qualitative and quantitative changes. These data underline the need for understanding the impact of perinatal factors on the gut microbiota also at low taxonomic levels, especially in the case of relevant microbial populations such as Bifidobacterium. The data obtained provide indications for the selection of the species best suited for the development of bifidobacteria-based products for different groups of neonates and will help to develop rational strategies for favoring a healthy early microbiota development when this process is challenged.     

Short communication: Effect of milk and milk containing Lactobacillus casei on the intestinal microbiota of mice

BALB/c mice were fed milk or Lactobacillus casei BL23 in milk for 14 d and fecal samples were collected at d 0, 4, and 7 as well as 1 and 8 d after the last administration. According to high-throughput DNA sequencing of the 16S rRNA genes extracted from the fecal microbiota, the bacterial diversity in the fecal samples of all mice increased over time. After 14 d of administration, the consumption of milk and milk containing L. casei BL23 resulted in distinct effects on the microbial composition in the intestine. Specifically, the proportions of bacteria in the Lactobacillaceae, Porphyromonadaceae, and Comamonadaceae were significantly higher in mice fed the L. casei BL23-milk culture compared with one or more of the other groups of mice. The relative amounts of Lachnospiraceae were higher and Streptococcaceae were lower in mice fed milk alone. The changes were not found at d 4 and 7 during milk and L. casei feeding and were no longer detected 8 d after administration was stopped. This study shows that consumption of milk or probiotic L. casei-containing milk results in non-overlapping, taxa-specific effects on the bacteria in the distal murine intestine.     

糖尿病患者肠道中4 种重要菌群的定量分析

目的：了解正常人群与Ⅱ型糖尿病人,以及Ⅱ型糖尿病人在是否有家族遗传、是否用药治疗和是否伴随肥胖等情况下肠道中拟杆菌、双歧杆菌、柔嫩梭菌和乳杆菌的含量与变化,为预防和治疗Ⅱ型糖尿病开辟新途径。方法：收集上述各类人群粪便样品（每组10 人）,分别提取样品中微生物的总DNA,运用荧光定量聚合酶链式反应（polymerase chain reaction,PCR）扩增4 种重要菌群的特异性片段,计算含量,分析上述菌群的变化差异。结果：正常人群肠道中的拟杆菌和双歧杆菌含量均显著低于Ⅱ型糖尿病人群;柔嫩梭菌和乳杆菌含量均显著高于后者。在Ⅱ型糖尿病人群中,用药患者组和肥胖＋高血糖患者组的肠道中拟杆菌和双歧杆菌含量均显著低于未用药患者组和单纯高血糖患者组,而柔嫩梭菌和乳杆菌含量均显著高于后者。有无Ⅱ型糖尿病遗传史对患者肠道内4 种菌群的含量均无显著影响。结论：4 种重要菌群与Ⅱ型糖尿病的发生和发展有密切关系;用药治疗后的患者肠道中4 种重要菌群含量比其他各组更接近正常人群;肥胖易加重Ⅱ型糖尿病病情;而有无Ⅱ型糖尿病家族遗传对这4 种菌群的含量几乎没有影响。