Persistence of probiotic strains in the gastrointestinal tract when administered as capsules, yoghurt, or cheese

Most clinical studies of probiotics use freeze-dried, powdered bacteria or bacteria packed in capsules. However, probiotics are commercially available in various food matrices, which may affect their persistence in the gastrointestinal tract. The objective of the study was to compare oral and faecal recovery during and after administration of a combination of Lactobacillus rhamnosus GG and LC705, Propionibacterium freudenreichii subsp. shermanii JS, and Bifidobacterium animalis subsp. lactis Bb12 as capsules, yoghurt, or cheese. This randomized, parallel-group, open-label trial (n = 36) included a 4-week run-in, 2-week intervention, and 3-week follow-up period. Participants consumed 10¹⁰ cfu/day of probiotic combination and provided saliva and faecal samples before, during, and after the intervention. Strain-specific real-time PCR was used to quantify the strains.L. rhamnosus GG was the only probiotic strain regularly recovered in saliva samples. During the intervention period it was recovered in the saliva of 88% of the volunteers at least once. No difference was found between the yoghurt and cheese groups. At the end of the intervention, L. rhamnosus GG and LC705 counts were high in faecal samples of all product groups (8.08 and 8.67 log₁₀ genome copies/g, respectively). There was no matrix effect on strain quantity in faeces or the recovery time after ceasing the intervention. For P. freudenreichii subsp. shermanii JS and B. animalis subsp. lactis Bb12, a matrix effect was found at the end of the intervention (P < 0.01 and P < 0.001, respectively) and in the recovery time during follow-up (P < 0.05 for both). Yoghurt yielded the highest faecal quantity of JS and Bb12 strains (8.01 and 9.89 log₁₀ genome copies/g, respectively). The results showed that the administration matrix did not influence the faecal quantity of lactobacilli, but affected faecal counts of propionibacteria and bifidobacteria that were lower when consumed in cheese. Thus, the consumption of probiotics in yoghurt matrix is highly suitable for studying potential health benefits and capsules provide a comparable means of administration when the viability of the strain in the capsule product is confirmed.     

Proteolysis in Cheese Curd as Affected by Subcellular Fractions from Lactococcus, Lactobacillus and Propionibacterium

Intracellular fraction (IF) and cell wall-associated fraction (CWAF) from lactococci, lactobacilli and propionibacteria were added to cheese curd slurries according to a mixture design. Response surface and estimation of Scheffe's polynomial revealed synergistic interaction effects between lactococci and lactobacilli on secondary proteolysis determined by an o-phthaldialdehyde assay. IFs from propionibacteria had a lower proteolytic activity than those from lactococci and lactobacilli used. Principal component analysis of capillary electropherograms of the trichloroacetic acid-soluble fraction, discriminated between slurries treated with IFs and CWAFs. It also showed an effect of proteolytic enzymes from the IF of propionibacteria together with IFs from lactococci and/or lactobacilli, on small changes in overall peptide profile.     

Effect of mesophilic lactobacilli and enterococci adjunct cultures on the final characteristics of a microfiltered milk Swiss-type cheese

The effect of four associations of adjunct cultures composed of mesophilic lactobacilli and enterococci, either solely or combined, on the microbiological, biochemical and sensory characteristics of Swiss-type cheese made using microfiltered cows’ milk and supplemented with propionibacteria was studied. The global pattern of growth was similar to that generally observed in raw milk cheese and interactions between microflora were highlighted during ripening. Enterococci, which negatively affected the survival of streptococci starters, seemed to play a limited role in the formation of volatile compounds, probably due to their low levels throughout ripening. On the contrary, mesophilic lactobacilli, which affected the evolution of propionibacteria, enterococci and L. delbrueckii subsp. lactis starter counts, modified free amino acid content, production of volatile compounds and organoleptic properties of mature cheese. This population appeared to be of major importance in the formation of cheese flavor as it was positively related to numerous potential flavor compounds such as alcohols and their corresponding esters, acetaldehyde and 4-methyl-4-heptanone. The original mesophilic lactobacilli present in milk could play an important role in the sensorial diversity of raw milk Swiss-type cheeses such as Comte.     

Assessment of microbial population dynamics during yoghurt and hard cheese fermentation and ripening by DNA population fingerprinting

The application of Terminal Restriction Fragment Length Polymorphism (T-RFLP) analysis of the small subunit ribosomal gene to examine the bacterial dynamics of relatively fast dairy fermentation processes is illustrated. All individual starter strains yielded characteristic terminal restriction fragments (TRFs). Growth independent T-RFLP analysis was used to identify and follow the dynamics of streptococci, lactobacilli, lactococci and propionibacteria in yoghurt and hard cheese production while samples had been stored frozen until analysis. Starter cultures could be differentiated and followed during production and ripening of hard cheese (Gouda type and Maasdam) as well as production of yoghurt. T-RFLP analysis enabled the characterization of bacterial population structure and dynamics between short time spans, i.e., within hours. Eight samples taken at different times within 5.5 h yoghurt fermentation revealed shifting relative signal intensities in TRF peaks between streptococci and lactobacilli, indicating a (semi) quantitative relation of the TRFLP signal with the actual numbers of bacteria.     

Removal of common Fusarium toxins in vitro by strains of Lactobacillus and Propionibacterium

This study was conducted to examine the ability of selected strains of Lactobacillus and Propionibacterium to remove common Fusarium toxins, trichothecenes, from liquid media. The trichothecenes studied were deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-AcDON), nivalenol (NIV), fusarenon (FX), diacetoxyscirpenol (DAS), T-2 toxin (T-2) and HT-2 toxin (HT-2). The Lactobacillus rhamnosus strain GG (LGG), Lactobacillus rhamnosus strain LC-705 (LC-705) and Propionibacterium freudenreichii ssp. shermanii JS (PJS) were incubated in PBS buffer containing 20 µg toxin ml -1 for 1h at 37°C, and after centrifugation the concentration of the toxins was measured in the supernatant fraction. Both viable and heat-killed forms of LGG and PJS were more efficient than LC-705 in removing the toxins from the liquid media. LGG and PJS removed four of the seven tested toxins (the removal varying from 18 to 93%) and LC-705 two toxins (10-64%). Of the toxins, 3-AcDON was not removed by any of the bacteria; HT-2 was removed by the non-viable LGG and also slightly by non-viable LC-705; DAS was removed by all three bacteria tested. Binding is postulated as the possible mechanism of the removal, since no difference was observed between the ability of viable and heat-killed bacteria in removing the trichothecenes, and no degradation products of the toxins were detected by gas chromatography (GC)-mass spectrometry (MS) analysis. It is concluded that significant differences exist in the ability of the bacteria to bind trichothecenes in vitro.     

益生菌Lactobacillus paracasei LC01对小鼠肠道菌群的调节作用

探讨益生菌Lactobacillus paracasei LC01对小鼠肠道菌群的调节作用。将32 只Balb/c小鼠随机分成4 组,分别作为对照组、灌胃LC01灭活菌组、灌胃LC01活菌组和活菌定殖组。干预2 周后,通过MiSeq高通量测序手段,分析不同处理条件下小鼠肠道菌群变化。结果表明：相对于对照组和灌胃灭活菌组,灌胃LC01活菌可以显著提高小鼠肠道内Lactobacillus属、Ruminococcaceae_UCG014属、Alistpes属的相对含量。进一步通过荧光定量聚合酶链式反应和群落聚类分析发现,灌胃LC01活菌不但可以显著提高小鼠肠道内乳酸菌LC01的相对含量,同时可以促进小鼠肠道中格氏乳杆菌、罗伊氏乳杆菌和唾液乳杆菌的增殖。在活菌定殖组中,停止灌胃2 周后,小鼠粪便中不能检测到菌株LC01,说明菌株LC01不能在小鼠肠道内长期定殖。以上结果表明菌株LC01对肠道小鼠菌群结构具有一定的调节作用,具有潜在的益生作用和应用价值。     

干酪乳杆菌LC2W对两种不同胃癌细胞的黏附作用

选取人胃癌细胞株MKN-45和HGC-27,采用形态学观察和显微计数的方法比较菌液浓度、悬浮液、pH值对干酪乳杆菌LC2W(Lactobacillus casei LC2W)黏附细胞的情况。结果表明,LC2W对两种细胞均具有黏附性,对MKN-45的黏附性更强;这种黏附作用具有菌悬液浓度依赖性。当加入的菌悬液浓度达到一定数量时(109 mL-1),平均每个细胞黏附的乳杆菌数量趋于饱和;悬浮介质对Lb.caseiLC2W的黏附性具有明显的影响,其中悬浮于耗尽培养上清液(SCS)的Lb.casei LC2W表现出最好的黏附性;悬浮介质的pH值对Lb.casei LC2W的黏附性也有影响,pH=4.0时黏附性最好。将不同悬浮介质的pH值调整到pH=7.0后,SCS仍能很好的促进LC2W的黏附作用,表明SCS中存在某种促进黏附的物质。由此推测,Lb.casei LC2W对胃癌细胞的黏附机理可能为黏附素与受体的特异性结合。     

Evaluation of the spiral plating method for the enumeration of microorganisms throughout the manufacturing and ripening of a raw goat's milk cheese

A statistical comparison of the spiral plate count (SPLPC) and the standard plate count (SPC) methods for enumeration of microorganisms in raw goat's milk cheese throughout its manufacturing and ripening was carried out. Enumeration of mesophiles, lactic acid bacteria (presumptive lactococci, presumptive leuconostocs, and presumptive lactobacilli), Micrococcaceae, Enterobacteriaceae, and molds and yeasts was carried out for milk, curd, and 2-, 5-, 10-, 17-, and 27-day-old cheeses. Average counts for the SPLPC and SPC methods differed by less than half of a log cycle for all microbial groups studied (range of difference, -0.1386 [mesophiles] to +0.4397 [presumptive lactobacilli]). The results of the SPLPC method compared favorably with the results of the SPC procedure for mesophiles, presumptive lactococci, presumptive leuconostocs, Enterobacteriaceae, and molds and yeasts (the variance between replicate platings was close to 0.005, and correlation coefficients were >0.9). Correlation coefficients were lower for Micrococcaceae (r = 0.824) and presumptive lactobacilli (r = 0.670). Analysis of variance showed that the plating method was a significant factor (P < 0.05) for presumptive lactobacilli counts. In general, results from the SPLPC method compared favorably with results from SPC procedure in the enumeration of microorganisms in goat cheese throughout its manufacturingand ripening processes. However, the suitability of the SPLPC method depends mainly on the microbial group studied.     

Influence of feeding fermented colostrum and Lactobacillus acidophilus on fecal flora of dairy calves

Twenty Holstein calves were assigned alternately at birth to diets of 1) fermented colostrum, 2) colostrum treated with 1% propionic acid, 3) whole milk, or 4) whole milk treated with Lactobacillus acidophilus (frozen concentrate culture) at 5 x 10(8) organisms per litter. Diets were fed once daily for 3 wk at 10% of birth weight as the sole source of nutritients. Fecal samples were collected at 0, 7, 14, and 21 days of age and analyzed for coliform and lactobacilli numbers. Fermented colostrum diets did not alter coliform counts in feces of healthy calves. Fecal coliform counts of calves fed L. acidophilus decreased with time. Average fecal lactobacilli counts were lower for the colostrum diets than milk diets. The apparent lowered incidence of scours frequently reported in calves fed fermented colostrum diets was not reflected in major changes in fecal microflora under the conditions of this study.     

A sensitive method for qualitative screening of bile salt hydrolase-active lactobacilli based on thin-layer chromatography

A sensitive protocol based on thin-layer chromatography (TLC) was developed to screen qualitatively bile salt hydrolase (BSH)-active lactobacilli. The sodium salts of glycocholic acid and taurocholic acid were used as substrates, and bacterial BSH activity was confirmed by detecting cholic acid as a product of the bile conjugates using a TLC assay with direct visual observation. Forty-five lactobacilli isolated from human fecal samples were tested for BSH activity by the TLC assay, a conventional plate assay, and a quantitative colorimetric assay. With the TLC and quantitative colorimetric assays, the same 24 BSH-positive strains were detected. No false-positive or false-negative results were detected by the TLC assay. However, only 20 BSH-positive strains were detected with the conventional plate assay. Compared with the conventional plate assay, the TLC assay is more sensitive for the detection of BSH activity of lactobacilli and, thus, more suitable for screening of BSH-active lactobacilli of human origin.     

Fermentation of feruloyl and non-feruloyl xylooligosaccharides by mixed fecal cultures of human and cow: a comparative study in vitro

For the first time, a comparative study was undertaken with respect to the fermentation of (a) monosaccharides (glucose, galactose, arabinose, and xylose); (b) disaccharide (lactulose); (c) Jerusalem artichoke; (d) xylooligosaccharides (XO); and (e) feruloyl xylooligosaccharides (FXO) by mixed fecal cultures of human and cow. Among monosaccharides, arabinose and xylose exhibited prebiotic properties, and among these arabinose was found to be a better substrate than xylose. Glucose and galactose did not have any impact with respect to either increase or decrease in different bacterial populations present in both human and cow feces and liberated very small amounts of SCFA indicating them to be the least prebiotic among all the substrates tried. Both lactulose and Jerusalem artichoke increased the lactobacilli and bifidobacteria in pooled fecal cultures of human and cow. Bovine fecal bacteria utilized XO and FXO more effectively than human fecal bacteria as indicated by relatively high levels of the cell wall–degrading enzyme activities. Growth of different bacterial populations was monitored by the fluorescent in situ hybridization method at 12 and 24 h. XO increased the growth of lactobacilli and bifidobacteria and decreased the growth of Bacteriodes and clostridia, whereas FXO increased the growth of lactobacilli in cow fecal cultures. In human fecal cultures, FXO promoted the growth of bifidobacteria, but to a lesser extent compared with cow fecal bacteria. Quantitative variations were observed with respect to the profile of short-chain fatty acids liberated in the fecal culture filtrates of human and cow grown on XO and FXO.     

Impact of consumption of probiotic lactobacilli-containing yogurt on microbial composition in human feces

An in vivo study was carried out to determine the effect of consuming probiotic lactobacilli-containing yogurt on the composition of microbiota in the human gut. Fifteen healthy adults ingested a daily serving of one of three commercial yogurts (two of the products contained a probiotic lactobacilli strain) for 20 days. Fecal samples at defined time points before, during, and after the period of yogurt ingestion were collected and analyzed. The fecal population of lactobacilli was determined by a culture-based method and subsequent colony PCR for the identification of species. Six predominant bacterial groups in the fecal samples were quantitatively determined based on a sequence-specific SSU rRNA cleavage method coupled with a suite of oligonucleotide probes, which was optimized for the target-specific detection of bacterial groups inhabiting human feces. In the ingestion period, one probiotic strain was detected in the feces of all five subjects who consumed the yogurt containing the strain, while the other strain was detected in three of another five subjects. The population levels of the two major groups (Bacteroides and Prevotella, and the Clostridium coccoides-Eubacterium rectale group) in the fecal samples tended to change in response to the ingestion but the change did not seem to be dependent on the product-specific property of each yogurt. These results suggest that the human fecal bacterial community could be altered by ingesting yogurt, although whether probiotic lactobacilli are present or absent in the yogurt does not seem to be a factor in this change.     

Impact of Bifidobacterium animalis subsp. lactis BB-12 and, Lactobacillus acidophilus LA-5-containing yoghurt, on fecal bacterial counts of healthy adults

This randomized, placebo-controlled, double blind, parallel dose-response study investigated the impact of 4-week commercial yoghurt consumption supplemented with Bifidobacterium animalis subsp. lactis (BB-12) and Lactobacillus acidophilus (LA-5) on fecal bacterial counts of healthy adults. Fifty-eight volunteers were randomly assigned to three different groups: 1. placebo (no probiotic, no starter and no green tea extract); 2. Yoptimal (10⁹ cfu/100 g of BB-12 and LA-5 and 40 mg of green tea extract) and 3. Yoptimal-10 (10¹⁰ cfu/100 g of BB-12, 10⁹ cfu/100 g of LA-5 and 40 mg of green tea extract). These yoghurt products also contained Lactobacillus delbrueckii subsp. bulgaricus (10⁷ cfu/100 g) and Streptococcus thermophilus (10¹⁰ cfu/100 g). The quantitative PCR (qPCR) results showed that there were significant increases (P = 0.02) in bifidobacteria counts with the Yoptimal treatment as compared to baseline. The fecal numbers of B. animalis subsp. lactis and LA-5 significantly increased in the two probiotic treatments compared to the placebo treatment. Viable counts of fecal lactobacilli were significantly higher (P = 0.05) and those of enterococci were significantly lower (P = 0.04) after the intervention when compared to placebo. No significant difference was observed between treatments in volunteers' weight, waist girth, blood pressure, fasting plasma triglyceride and HDL-C concentrations, as well as cholesterol/HDL-cholesterol ratio. However, a significant increase in plasma cholesterol levels was observed in the placebo group (P = 0.0018) but the levels remained stable in the two probiotic yoghurt groups. These results show that probiotic strains supplemented in the form of yoghurt remain active during gut transit and are associated with an increase in beneficial bacteria and a reduction in potentially pathogenic bacteria. This trial was registered at clinicaltrials.gov as NCT00730626.     

西藏芜菁及其加工制品增强人体低氧耐受性的实验研究

以西藏芜菁的块茎及其加工制品（芜菁脆片和芜菁饮料）为实验材料,选市售的抗缺氧药物红景天胶囊为阳性对照物,同时设淀粉胶囊作为安慰剂,进行了为期21 d的人体试食试验。实验期间受试者每周定时进行低氧耐受性测试,记录受试者在低氧环境中的不耐受主诉、血氧饱和度及心率,同时检测血清超氧化物歧化酶（superoxide dismutase,SOD）、过氧化氢酶（catalase,CAT）酶活性及丙二醛（malondialdehyde,MDA）含量等指标。结果表明：芜菁及其加工制品能有效缓解人体的低氧反应症状;各实验组（安慰剂除外）实验后的心率均小于实验前。在增强人体抗氧化能力方面,阳性对照组和芜菁脆片组的受试者连续服用7 d和14 d后血清SOD酶活性较实验前得到显著升高（P＜0.05）;第21天时,与安慰剂组相比,各实验组的人体血清CAT酶活性均有显著升高（P＜0.05）,而MDA含量则显著降低（P＜0.05）。综上所述,芜菁及其加工制品均具有一定的抗缺氧作用,尤其是芜菁脆片很好地保留了芜菁鲜品中的有效成分,其生物学功效可与红景天媲美,是一种具有良好开发前景的抵御高原反应的即食休闲食品。     

Antifungal effect of dairy propionibacteria--contribution of organic acids

Large amounts of food and feed are lost every year due to spoilage by moulds and yeasts. Biopreservation, i.e. the use of microorganisms as preservatives instead of chemicals, has gained increased interest. Lactic acid bacteria and propionibacteria might be particularly useful due to their important role in many food fermentations. Knowledge of the antifungal effects of the organic acids produced by these bacteria is necessary to understand their inhibitory activity. We evaluated the antifungal activity of the type strains of five dairy propionibacteria, Propionibacterium acidipropionici, P. jensenii, P. thoenii, P. freudenreichii subsp. freudenreichii and P. freudenreichii subsp. shermanii against eight food- and feedborne moulds and yeasts. A dual culture system assayed the inhibitory activity on three different agar media, sodium lactate (SL), de Man Rogosa Sharp (MRS) and MRS without acetate (MRS-ac). The amounts of organic acids produced during growth of propionibacteria in liquid SL, MRS and MRS-ac were also determined. The minimal inhibitory concentration (MIC) values of propionic, acetic and lactic acid were established for all fungi at pH 3, 5 and 7. Propionic acid, followed by acetic acid, was the most potent antifungal acid. Inhibition at pH 7 generally required concentrations above 500 mM for all three acids, at pH 5 the MIC values for propionic and acetic acids were 20-120 mM and above 500 mM for lactic acid. At pH 3, the MIC values were, with one exception, below 10 mM for both propionic and acetic acid and above 160 mM for lactic acid. The yeast Pichia anomala was the fungus most resistant to organic acids. The propionibacteria exhibited a pronounced species variation in antifungal activity on MRS (+/-acetate) agar, with P. thoenii being the most potent. Four of the five propionibacteria species produced more propionic and acetic acid in liquid SL medium than in MRS (+/-acetate) broth. However, when SL agar was used as the growth medium, none of the propionibacteria inhibited fungal growth.     

In vitro activity on human gut bacteria of murta leaf extracts (Ugni molinae turcz.), a native plant from southern Chile

Despite the fact that murta infusions have been used to treat gut/urinary infections by native Chileans for centuries, the mechanisms promoting such effects still remain unclear. As a first attempt to unravel these mechanisms, human fecal samples were incubated in a medium containing water extract of murta leaves (ML) and the growth of different bacterial groups was evaluated. Control incubations were made in media containing fructooligosaccharides (FOS) and glucose as a carbon source. Phenolic compounds in the ML extract, likely promoters of bioactivity, were identified by HPLC-DAD-MS(n) . Concentrations (log(10) CFU/mL) of bifidobacteria and lactobacilli in media containing the extract and FOS were 7.33 ± 0.05/4.95 ± 0.20 and 6.44 ± 0.22/6.05 ± 0.06, respectively. Clostridia, anaerobes and Enterobacteriaceae grew to a similar extent in media containing murta extract and FOS. In vitro tests (disk diffusion) showed that Gram-positive (Bacillus and Paenibacillaceae) and Gram-negative (Enterobacteriaceae) bacteria isolated from fecal samples were sensitive to both water and 50/50 ethanol/water extracts of ML (28.4 μg gallic acid equivalents). At this concentration, the antimicrobial activity of ML extracts was significantly (P < 0.05) lower than that of penicillin (10 U), whereas the difference between activity of ML extracts and gentamicine (10 μg) was no significant (P > 0.05). No evidence of dependency between the antimicrobial activity of ML extracts and the enzymatic capability of the sensitive strains was found.     

Dietary cheese whey protein protects rats against mild dextran sulfate sodium-induced colitis: Role of mucin and microbiota

Data from the literature suggest that the availability of the amino acids threonine, cysteine, or both, is limiting for mucin synthesis under conditions of chronic inflammatory bowel disease. Unlike casein, cheese whey protein is rich in these amino acids. The protective effect of cheese whey protein was examined using dextran sulfate sodium (DSS)-induced inflammation of the large intestine in rats that were fed a diet containing casein, cheese whey protein, or casein supplemented with threonine and cysteine. The clinical markers diarrhea and fecal blood were determined using biochemical assays, and gene expression of inflammation markers was used to quantify inflammation. The effect of dairy protein on mucin production was determined by gene expression of rat mucin 2 (MUC2) and by quantifying fecal mucin excretion. Fecal lactobacilli and bifidobacteria were determined using quantitative PCR. Dietary cheese whey protein reduced DSS-induced gene expression of the inflammation markers interleukin 1β, calprotectin, and inducible nitric oxide synthase, and diminished the clinical symptoms diarrhea and fecal blood loss. Moreover, cheese whey protein increased fecal mucin secretion without affecting gene expression of MUC2, suggesting enhanced mucin synthesis. In addition, cheese whey protein increased fecal lactobacilli and bifidobacteria counts. Supplementation of threonine and cysteine showed comparable effects. In conclusion, cheese whey protein protected rats against DSS-induced gut inflammation. This can most likely be explained by its threonine and cysteine content. Protection can be the result of both the stimulation of intestinal mucin synthesis and modification of microflora composition.     

Effect of a monensin-controlled release capsule on cow health and reproductive performance

Dry cows and pregnant heifers from 25 farms near Guelph, Ontario, Canada were enrolled in a large, double-blind, randomized clinical trial designed to evaluate the impact of monensin on energy metabolism, health, and production. A total of 503 cows was given monensin in controlled-release capsules, and 507 cows were administered placebo capsules 3 wk before expected calving date. The effects of treatment on health were evaluated using a logistic regression model. Treatment with monensin significantly reduced the incidence of abomasal displacement (OR = 0.41-0.84) and multiple illnesses (OR = 0.38-0.89). Monensin treatment tended to reduce the incidence of clinical ketosis (P = 0.11) and the risk of being culled (P = 0.09) in the first 94 d of lactation. Reproductive performance was analyzed with both a logistic regression model for conception rate and a survival analysis for days to first breeding and days from calving to conception. Treatment with monensin had no significant effect on any measure of reproductive performance.     

低聚木糖对慢性腹泻症状的改善作用

本研究旨在探讨低聚木糖（xylooligosaccharides,XOS）对慢性腹泻的改善作用。基于随机平行对照实验原则,将临床慢性腹泻患者随机分为3 组：安慰剂组（CK组,每天3 g麦芽糊精）、低剂量组（3X组,每天3 g XOS）和高剂量组（6X组,每天6 g XOS）;干预4 周后,记录患者腹泻症状评分,测定患者血清生化指标、粪便中短链脂肪酸和肠道菌群的变化。随后用3 组干预后患者粪菌液分别灌胃3 组小鼠,连续灌胃24 d,测定其肠道通透性及炎症因子水平。结果表明,与安慰剂组相比,XOS干预患者4 周后,患者腹泻症状和脂代谢有改善趋势,粪便中丁酸含量提高。与干预前相比,XOS干预患者粪便中Blautia、Bifidobacterium、Lachnospiraceae_unclassified等物种相对丰度增加,Prevotella等菌属相对丰度下降。与灌胃安慰剂组患者粪菌液的小鼠相比,灌胃XOS干预组患者粪菌液的小鼠肠道屏障趋于稳定状态,结肠炎症显著缓解,肠道通透性相关指标明显改善,血清中D-乳酸水平显著降低（P＜0.05、P＜0.01）,闭锁小带蛋白（zonula occludens-1,ZO-1）和紧密连接蛋白Occludin表达量增加。综上,XOS在慢性腹泻的改善中发挥了较好的效果,具有潜在缓解慢性腹泻的作用。     

Shelf-life of vacuum-packed cooked ring sausages at different chill temperatures

Microbiological and sensory changes in 313 vacuum-packed cooked ring sausages from 28 different production runs and stored at 2, 4, 8 or 12 degrees C were monitored as a function of time. The sensory scores started to decrease at a level of approx. 10(7) lactobacilli/g. The judges began considering the samples unfit for human consumption when the lactobacilli counts were between 10(7) and 10(8) cfu/g; above a level of 10(8) cfu/g most of the samples were deemed unfit. At 2 degrees C, however, spoilage did not always seem to be microbiological, and four out of six different production runs were deemed unfit without any marked increase in microbial counts. In such cases, the judges described the sensory defects as a 'musty' rather than a sour aroma and taste. The sausages were deemed unfit when the lactobacilli were in a stationary growth phase which was considerably later than the point when the bacterial counts exceeded 10(7) cfu/g. The mean length of this delay was 30, 19, 16 and 7 days at 2, 4, 8 and 12 degrees C, respectively. The average shelf-lives were 55, 43, 29 and 17 days at 2, 4, 8 and 12 degrees C, respectively. The dependence of shelf-life on temperature can be formulated as follows: Shelf-life = 10(1.835 - 0.048 X temperature) The maximal shelf-life of this product, including nonmicrobiological spoilage, is assessed as approx. 10-11 weeks. A lactobacilli count greater than 10(7) cfu/g indicates that either the spoilage process has started or the product is already spoiled. When the lactobacilli count exceeds 10(8) cfu/g it is highly probable that the sausage sample is unacceptable.