免疫婴儿乳粉的安全毒理学评定

以24株人肠道病原菌(包括病原性大肠杆菌12株、沙门氏菌8株、志贺氏菌3株、小肠结肠炎耶尔森氏菌1株)作为抗原,对乳牛进行系统免疫,免疫乳与非免疫乳乳中IgG含量无显著差异。系统免疫并不增加乳中IgG的含量,但IgG的抗体特异性大大增强,所得的免疫初乳中乳抗体对24种不同病原菌的凝集价为28～212,为普通初乳中乳抗体凝集价的32～256倍。以免疫初乳作为原料制备免疫初乳粉并添加到婴儿乳粉中制成免疫婴儿乳粉。婴儿免疫乳粉LD50>10g/kg;Ames试验及小鼠骨髓微核试验和小鼠精子畸变试验表明婴儿免疫乳粉无致畸变作用。大鼠30d喂养试验表明,婴儿免疫乳粉对大鼠生长无不良影响,大鼠血液指标正常,病理学检查未见任何病变。     

鼠李糖乳杆菌DHC32对小鼠肠道菌群和肠道黏膜免疫的影响

本文主要研究了鼠李糖乳杆菌DHC32对正常小鼠肠道菌群和肠道黏膜免疫的影响。结果表明,DHC32降低了粪便中氨、水分及pH值,提高了小鼠粪便菌群营养代谢率。与对照组相比,DHC32处理组小鼠粪便中乳杆菌、双歧杆菌等有益菌的数量显著提高,而拟杆菌、肠杆菌等有害菌的数量显著下降,并且短链脂肪酸的含量大幅提高。本文还证明鼠李糖乳杆菌DHC32通过提高My D88、IL-23及IL-22等免疫因子和抗菌蛋白RegIIIγ,黏附蛋白MUC1、MUC2基因的表达,来稳固黏膜屏障功能,增强宿主的免疫机能而改善动物的肠道健康。     

东北酸菜中产胞外多糖乳杆菌的分离筛选及其在酸乳中的应用

从东北酸菜中分离出97株乳杆茵菌株,并筛选出两株产酸快、产胞外多糖(EPS)的乳杆茵2号菌株和9号菌株.初步鉴定前者为干酪乳杆茵,后者为植物乳杆茵.在发酵剂(保加利亚乳杆菌和嗜热链球茵,LS)中混入等量的菌株2或9培养物(脱脂乳),所制备的酸乳中EPS质量浓度达34 mg/L.活茵总数可迭2×108mL-1以上.酸乳物性测定结果显示,产EPS菌株发酵的酸乳弹性好,相对粘度高,持水力强,乳清析出少.感官评定结果显示,与没有添加产EPS茵株发酵的酸乳相比,产EPS茵株LS+2与LS+9组合发酵酸乳风味分值较高,组织状态的分值差别较大,而酸度上没有显著差异.与物性测定结果一致.     

免疫初乳中乳抗体对大肠杆菌和沙门氏菌所致小鼠腹泻的被动免疫保护

以24株人肠道病原菌(包括病原性大肠杆菌12株、沙门氏菌8株、志贺氏菌3株、小肠结肠炎耶尔森氏菌1株)作为抗原，对乳牛进行系统免疫，免疫乳与非免疫乳乳中IgG含量无显著差异。系统免疫并不增加乳中IgG的含量，但IgG的抗体特异性大大增强，所得的免疫初乳中乳抗体对24种不同病原菌的凝集价为2^8-2^12，为普通初乳中乳抗体凝集价的32—256倍。采用硫酸胺盐析从免疫初乳中分离乳抗体，免疫初乳中的特异性IgG乳抗体较普通初乳中的IgG可显著抑制大肠杆菌和沙门氏菌的生长。动物试验表明，免疫乳中特异性的IgG对大肠杆菌和沙门氏菌所致小鼠腹泻具有很好的保护作用，而普通乳中非特异性的IgG则无此作用，这是由于IgG的特异性所决定。     

山羊乳与牛乳配方乳粉的致敏性比较

山羊乳是一类营养丰富且易于消化的乳品,因其营养成分与牛乳相近而受到广泛关注。本研究通过蛋白质潜在致敏性树状评估策略（生物信息学、消化稳定性、血清学研究、动物模型）对山羊乳配方乳粉的致敏性进行评价,并研究山羊乳配方乳粉与牛乳过敏患者血清中免疫球蛋白（immunoglobulin,Ig）E之间存在的交叉反应性。结果表明,山羊乳配方乳粉的167 种蛋白质中,β-乳球蛋白和α-乳清蛋白含量显著低于牛乳配方乳粉（P＜0.05）,山羊乳配方乳粉在模拟胃液中更易消化,且与过敏患者血清的结合能力弱于牛乳配方乳粉,动物模型结果表明山羊乳配方乳粉致敏小鼠体温下降幅度、临床过敏症状评分、血清中特异性IgE和IgG1抗体水平、血浆中组胺水平、血管渗透性及组织炎症程度等均显著低于牛乳配方乳粉组致敏小鼠（P＜0.05）,交叉反应性结果表明山羊乳配方乳粉能与牛乳过敏患者血清结合,但结合能力明显低于牛乳配方乳粉。结论：与牛乳配方乳粉相比,山羊乳配方乳粉的致敏性较弱,可作为牛乳的替代品,降低牛乳过敏性疾病的发病风险。     

瑞士乳杆菌对小鼠肠道粘膜免疫应答及细胞因子的影响

探讨灌胃瑞士乳杆菌(Lactobacillus helveticus TS206)发酵制品对健康Balb/c 小鼠肠道黏膜免疫应答及细胞因子的影响。本实验将Balb/c 小鼠分为空白对照组、生理盐水组和瑞士乳杆菌组3 组,瑞士乳杆菌组的灌胃剂量为108CFU/mL,分别在灌胃后0 、1 、2、3 、4 、6 、8、12、24h 处死小鼠得到小肠组织样品,利用RIA 法和ELISA 法测得各组小鼠肠道黏膜SIgA 和细胞因子的含量,并作统计分析。结论：灌胃组小鼠肠道黏膜中SIgA 的含量与生理盐水对照组相比差异极显著(P ＜ 0.01);灌胃组小鼠肠道黏膜中IL-2 和IFN-γ的含量与生理盐水对照组相比具有显著性差异(P ＜ 0.05);灌胃组IL-4 与生理盐水对照组相比含量明显升高(P ＜ 0.05);而灌胃组和生理盐水对照组相比IL-6 的表达水平后期差异不显著(P ＞ 0.05);IFN-γ/IL-4 的值处于相互拮抗的状况,表明瑞士乳杆菌对健康小鼠具有维护肠黏膜免疫平衡的作用,也对保持Th1/Th2 平衡状态具有明显的作用,即证实瑞士乳杆菌作为益生菌具有调控小鼠肠道黏膜免疫应答的功效。     

含24种乳抗体免疫乳的制备

以24株人肠道病原菌（包括病原性大肠杆菌12株、沙门氏菌8株、志贺氏菌3株、小肠结肠炎耶尔森氏菌1株）作为抗原，对乳牛进行系统免疫，免疫乳与非免疫乳乳中IgG含量无显著差异。系统免疫并不增加乳中IgG的含量，但IgG的抗体特异性大大增强，所得的免疫初乳中乳抗体对24种不同病原菌的凝集价为2^8-2^12，为普通初乳中乳抗体凝集价的32－256倍；免疫常乳中乳抗体对24种不同病原菌的凝集价为2^5-2^8，为普通乳中乳抗体凝集价的8－128倍。     

乳铁蛋白对小鼠肠道微生物区系的影响研究

目的:分析灌胃乳铁蛋白对小鼠肠道内一些具有代表性的微生物种群的变化规律及其特征,进一步了解乳铁蛋白对肠道菌群结构的作用,为开发具有肠道健康维护的功能性食品提供科学依据.方法:将小鼠饲养一周后随机分为5组,对照组,安慰剂组,低、中、高乳铁蛋白组,对照组正常饮食,安慰剂组灌胃生理盐水0.2mL,实验组灌胃不同剂量(1、10、100mg/(kg·d))的乳铁蛋白溶液,连续喂养14d.利用传统培养技术分别对小鼠肠道粪便菌群进行测定,通过数据分析了解小鼠肠道菌群的变化.结果表明喂食乳铁蛋白(10mg/(kg·d))后小鼠肠道中乳酸菌,双歧杆菌数量显著升高(p＜0.05);而肠道中的肠杆菌和肠球菌的数量明显降低(p＜0.05).说明一定剂量的乳铁蛋白(10mg/(kg·d))对肠道中的益生菌和常驻共生菌的生长具有显著的促进作用,而对肠杆菌,肠球菌等细菌均有一定的抑制作用.     

不同剂量植物乳杆菌对小鼠肠道菌群的影响

植物乳杆菌是人体肠道中的益生菌,可自发酵乳肉制品中分离。本实验将处于正常生理状态下的小鼠分为4组,通过灌喂不同剂量的植物乳杆菌X3-2B来探讨其对小鼠肠道各类微生物菌群的影响。结果表明,灌喂植物乳杆菌X3-2B时,小鼠对饲料的利用率优于对照组,且小鼠粪便p H低于对照组(高剂量组最低)。灌喂乳酸菌的小鼠其肠道内乳酸菌与双歧杆菌数显著(p<0.05)高于对照组。高剂量组与中剂量组小鼠在灌喂14d时小鼠肠道内大肠杆菌数无显著差异,但总肠杆菌和肠球菌数显著低于灌喂第1d(p<0.05),即植物乳杆菌X3-2B对肠道内致病菌有一定的抑制作用,对肠道微生物区系有一定的调节作用。     

副干酪乳杆菌L9对小鼠肠道短链脂肪酸含量的影响

目的:评价副干酪乳杆菌L9对健康小鼠肠道内短链脂肪酸(short chain fatty acids,SCFAs)含量的影响。方法:选取3周龄BALB/c雄性小鼠50只,随机分为副干酪乳杆菌L9高、中、低剂量组、阳性对照组(商业菌株Lactobacillus casei strain Shirota)、阴性对照(生理盐水)组共5组,以0.2 m L/d的剂量连续灌胃小鼠21 d并收集第0、21天小鼠粪便。第21天处死小鼠后搜集收集盲肠内容物,利用气相色谱仪观察内容物中SCFAs的变化,并且利用实时定量聚合酶链式反应(real-time polymerase chain reaction,RT-PCR)分析肠道菌群数量的变化。结果:与第0天相比,第21天小鼠粪便中SCFAs中乙酸、丙酸含量极显著上升(P0.01),Bifidobaceterium spp.、Lactobacillus属数量和丁酰辅酶A转移酶(butyryl-Co A:acetate Co A-transferase,CoAT)基因的表达量显著升高(P0.05);盲肠内容物中,低剂量组和高剂量组中的Bifidobaceterium spp.数量和CoAT基因表达量分别与第0天和阴性对照组相比有显著差异(P0.05)。实验结果表明:副干酪乳杆菌L9可以调节肠道菌群,促进健康小鼠肠道中SCFAs的产生,并且与商业菌株Lactobacillus casei strain Shirota相比,副干酪乳杆菌L9对肠道内丙酸、丁酸的含量和肠道中Bifidobaceterium spp.的数量有更好的促进作用。     

胃肠道调理专用奶粉对便秘小鼠肠道动力和菌群组成的影响

目的:研究胃肠道调理专用奶粉润肠通便作用及对肠道菌群的影响。方法:自制胃肠道调理专用奶粉后,以KM小鼠为实验对象,采用盐酸咯哌丁胺建立小鼠便秘模型,以小鼠一般状态、小肠推进率、排便情况、小肠组织观察为考察指标结合16S rRNA基因测序分析小鼠粪便,考察其对小鼠的润肠通便作用及肠道菌群的影响。结果:胃肠道调理专用奶粉各剂量能够显著（P<0.05）缩短小鼠首粒排便时间、显著（P<0.05）提高小鼠小肠推进率,但未对小鼠小肠杯状细胞结构造成破坏,且随着剂量的提升,奶粉的胃肠调理效果越好。胃肠道调理专用奶粉虽然没有明显改变肠道微生物的丰富度和均匀度,但是能够明显改变小鼠的肠道微生态结构组成,显著（P<0.05）增加某些益生菌Lactobacillus、Bifidobacterium的相对丰度。结论:胃肠道调理专用奶粉能够影响肠道运动,调节肠道菌群结构从而改善洛哌丁胺引起的便秘。     

Ingestion of yogurt containing Lactobacillus acidophilus and Bifidobacterium to potentiate immunoglobulin A responses to cholera toxin in mice

Lactic acid bacteria have been reported to have benefits for the prevention and treatment of some forms of diarrhea and related conditions. To determine whether these effects might involve direct stimulation of the gastrointestinal immune response, we administered yogurt to try to enhance mucosal and systemic antibodies against an orally presented immunogen, cholera toxin. Yogurts were manufactured with starter cultures containing different species and strains of lactic acid bacteria. Mice were fed these yogurts for 3 wk, during which they were also orally immunized twice with 10 micrograms of cholera toxin. Blood was collected on d 0 and 21, and fecal pellets were collected weekly. Mice that were immunized orally with cholera toxin responded by producing specific intestinal and serum immunoglobulin (Ig)A anti-cholera toxin. Antibody responses of the IgA isotype were significantly increased in mice fed yogurts made with starters containing the conventional yogurt bacteria Lactobacillus bulgaricus and Streptococcus thermophilus supplemented with Lactobacillus acidophilus, Bifidobacterium bifidum, and Bifidobacterium infantis. Yogurt that was manufactured with starters containing only conventional yogurt bacteria produced less IgA anti-cholera toxin than did the control group fed nonfat dry milk. Although strong responses were also observed for IgG anti-cholera toxin in serum, the responses did not differ among groups. Thus, administration of yogurt supplemented with L. acidophilus and Bifidobacterium spp. enhanced mucosal and systemic IgA responses to the cholera toxin immunogen.     

骆驼乳对慢性酒精性肝损伤小鼠肠道菌群的影响

目的:探究骆驼乳对慢性酒精性肝损伤小鼠肠道菌群多样性及结构的影响。将雄性C57BL/6NCr小鼠随机分为4组:对照组(Con,n=6)、模型组(Et,n=6)、骆驼乳剂量组(EtCM,n=6)和牛乳剂量组(EtNM,n=6);实验期为8周,前4周饲喂Lieber-DeCarli液体饲料(含对照),后4周在饲喂Lieber-DeCarli液体饲料的基础上,灌胃相应的乳或生理盐水。灌胃结束后,按照5 g/kg剂量一次性灌胃31.5%酒精溶液,建立NIAAA模型。检测血清LPS含量,并在无菌条件下取小鼠结肠粪便,进行16S rRNA测序,分析肠道菌群α多样性、β多样性及基于门、属水平的物种结构。血清指标结果显示,EtCM组和EtNM组小鼠血清LPS显著降低(P<0.01)。16S r RNA测序结果表明,骆驼乳和牛乳能显著提高ALD小鼠结肠肠道菌群的丰度和均匀度,更好地调整肠道菌群结构,其中骆驼乳较牛乳显示出更好的α多样性。在门水平上,骆驼乳和牛乳显著提高拟杆菌门的丰度,降低厚壁菌门的丰度。在属水平上,骆驼乳和牛乳显著提高副拟杆菌属、拟杆菌属、阿克曼菌属的丰度,降低瘤胃菌科下的未知属Ruminococcaceae_UCG-013丰度。其中,骆驼乳的有益菌丰度较牛乳高出9%。结论:骆驼乳通过改变ALD小鼠肠道菌群环境,来调节肠道菌群结构,可作为调节肠道菌群的功能性乳制品,可预防慢性ALD引起的肠道屏障功能障碍。     

Fermented milk supplemented with probiotics and prebiotics can effectively alter the intestinal microbiota and immunity of host animals

Fermented milk supplemented with 2 probiotic strains, Bifidobacterium lactis Bi-07 and Lactobacillus acidophilus NCFM, and a prebiotic, isomaltooligosaccharide, was orally administered to 100 healthy adults at 480g/d for 2 wk in a randomized controlled trial. The fecal bacterial compositions of these subjects were examined by culture before and after the intervention. The same fermented milk was also orally fed to BALB/c mice, and immune as well as fecal bacteria analyses were conducted using the same culturing methods. After the intervention, increases in fecal bifidobacteria and lactobacilli were observed among the subjects compared with the subjects in the control group. In contrast, after the intervention, fecal enterobacilli were significantly decreased in the test group compared with the control group. The same effects on the composition of the intestinal microbiota were observed in mice. Furthermore, the tested mice were found to have significantly increased delayed-type hypersensitivity, plaque-forming cells, and half-hemolysis values after the intervention with the fermented milk. In summary, the synbiotic fermented milk containing probiotics and a prebiotic may contribute to improve intestinal health and may have a positive effect on the humoral and cell-mediated immunity of host animals.     

北京地区不同喂养方式婴幼儿肠道菌群多样性分析

目的 探讨不同喂养方式对0~3岁婴幼儿肠道微生物的影响,以及喂养方式引起的肠道菌群变化与婴幼儿的年龄和分娩方式之间的相关性。方法 选取221例0~3岁北京地区健康婴幼儿肠道粪便样品,按照哺乳方式,将这些样品分为4组：纯母乳组（B）、纯奶粉组（F）、以母乳为主,奶粉和辅食为辅组（BP）、以奶粉为主,辅食为辅（NB）。其中B组44例、F组29例、BP组108例、NB组40例。利用16s微生物多样性测序技术分析了不同喂养方式对婴幼儿肠道微生物的丰度影响。结果 在物种组成上,没有摄入母乳的NB组,门水平上优势菌的含量变化明显。厚壁菌门、变形菌门、放线菌门和拟杆菌门四大优势菌的总含量明显低于B组、BP组和F组。在属水平上B组、BP组和F组相互比较,F组双歧杆菌属和拟杆菌属的含量显著下降,但粪杆菌属、罕见小球菌属等厌氧菌的含量升高。主成分分析的结果表明,NB组和B组的距离最远,F组与BP组的距离最近。F组与B组的差异要大于BP组与B组的差异,说明,与纯母乳喂养的婴幼儿相比,奶粉喂养的婴幼儿其肠道微生物的变化要大于辅以母乳摄入的婴幼儿。同时,关联性分析表明,分娩方式与喂养方式和婴幼儿年龄在影响肠道菌群的丰富度上密切相关。结论 出生前3年的饮食组成和模式会显著影响肠道微生物群的多样性,这种多样性变化与婴幼儿的年龄和分娩方式均具有密切的相关性。     

Effect of fermented milk containing probiotic bacteria in the prevention of an enteroinvasive Escherichia coli infection in mice

This study investigated the protective capacity of the oral administration of fermented milk containing the probiotic strains; Lactobacillus casei, Lb. delbrueckii subsp. bulgaricus and Streptococcus thermophilus, against enteroinvasive Escherichia coli infection in a murine (BALB/ c mice) model. Mice were fed for 2, 5 or 7 consecutive days with fermented milk diluted to a concentration of viable Lb. casei, Lb. delbrueckii subsp. bulgaricus and Strep. thermophilus of 10(7) cfu/ml. Phagocytic activity of peritoneal macrophages and the number of IgA+ cells in small and large intestine were determined at the end of the feeding periods. For the preventive effect against Esch. coli, animals were fed for 5 days (selected dose). Mice were challenged with an infective dose of enteroinvasive Esch. coli of 10(8) cfu/mouse. The colonization of liver and spleen and the secretory IgA specific for the pathogen in the intestinal fluid were determined (ELISA test). Results showed that the unspecific immune response enhanced itself after 5 consecutive days of the administration of this fermented milk (increase in the percentage of phagocytosis and number of IgA+ cells in the small intestine). Treated animals showed less Esch. coli colonization of liver than control mice and a higher secretory anti-Esch. coli IgA in the intestinal fluids. These results suggest that the protection against enteroinvasive Esch. coli infection observed for the fermented milk containing probiotic bacteria may be associated with an enhance of the intestinal mucosa immunity.     

Prebiotics: A Potential Treatment Strategy for the Chemotherapy-damaged Gut?

Mucositis, characterized by ulcerative lesions along the alimentary tract, is a common consequence of many chemotherapy regimens. Chemotherapy negatively disrupts the intestinal microbiota, resulting in increased numbers of potentially pathogenic bacteria, such as Clostridia and Enterobacteriaceae, and decreased numbers of “beneficial” bacteria, such as Lactobacilli and Bifidobacteria. Agents capable of restoring homeostasis in the bowel microbiota could, therefore, be applicable to mucositis. Prebiotics are indigestible compounds, commonly oligosaccharides, that seek to reverse chemotherapy-induced intestinal dysbiosis through selective colonization of the intestinal microbiota by probiotic bacteria. In addition, evidence is emerging that certain prebiotics contribute to nutrient digestibility and absorption, modulate intestinal barrier function through effects on mucin expression, and also modify mucosal immune responses, possibly via inflammasome-mediated processes. This review examines the known mechanisms of prebiotic action, and explores their potential for reducing the severity of chemotherapy-induced mucositis in the intestine.     

黑米对小鼠肠道菌群的影响

研究黑米对小鼠肠道菌群及其相关指标的影响。24只昆明(KM)小鼠随机分成4个组:组1(饲喂基础饲料)、组2(饲喂30%黑米和70%基础饲料的混合饲料)、组3(饲喂60%黑米和40%基础饲料的混合饲料)、组4(饲喂90%黑米和10%基础饲料的混合饲料)。第6周收集小鼠粪便测定粪便含水率,第42天测定血清总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)以及肠道双歧杆菌、乳酸杆菌、肠球菌和肠杆菌数量。结果表明,与组1小鼠相比,摄入黑米后,试验组小鼠结肠双歧杆菌数和乳酸杆菌数显著上升(P<0.05),组2最高,其次是组3和组4;肠杆菌科数显著下降(P<0.05),组2最低,组4次之;组2和组4肠球菌数显著低于组1(P<0.05)。试验组小鼠的血清HDL-C和HDL-C/TC都显著高于组1(P<0.05),组4的HDL-C最高,组2的HDL-C/TC最高。表明摄入全谷黑米可以提高小鼠肠道益生菌数量,全谷黑米对肠道具有益生作用。     

健康成人粪便中双歧杆菌的分离鉴定及其发酵乳体内特性研究

为筛选具有潜在益生功能的双歧杆菌,采用MRS培养基,结合菌落形态观察和16S rDNA基因序列同源性分析,从上海地区健康的成人粪便中分离鉴定双歧杆菌,通过7 L发酵罐发酵试验筛选优良菌株,并利用其制备发酵乳,对发酵乳的体内特性进行研究。结果表明,分离并鉴定到3株长双歧杆菌（Bifidobacterium longum）（编号为P-1、P-2和P-3）,其中菌株P-2制备的发酵乳在稳定期活菌数最高,遗传稳定性好,被确定为优良菌株,并被命名为DD98。菌株DD98制备的发酵乳能显著增加小鼠胃肠道内有益菌、减少有害菌的表达（P＜0.05）,具有调节肠道菌群的作用。此外,能维持小鼠正常的体质量、血常规指数,有利于小鼠脏器的生长。     

基于三代测序技术分析新疆鲜马奶和酸马奶中乳酸菌菌群结构

传统酸马奶受制作环境和工艺的影响,其菌群结构有所不同.本文运用Pacbio SMRT测序技术,基于乳酸菌特异性引物对新疆部分地区酸马奶和鲜马奶的乳酸菌菌群进行研究,结果表明:鲜马奶中乳酸菌丰富,以瑞士乳杆菌(35.52％)、唾液链球菌(10.11％)、乳酸乳球菌(9.49％)、嗜热链球菌(5.96％)以及德氏乳杆菌(5...