伯顿毕赤酵母菌在酸性及胆盐环境的抗逆性研究

目的探究伯顿毕赤酵母菌在酸性及胆盐环境下的益生特性,为研制益生菌制剂提供理论依据。方法采用稀释平板计数法,在pH 1.0、1.5、2.0、2.5、3.0和胆盐浓度0.1%、0.2%、0.3%、0.4%的PDA液体培养基以及人工胃液作用0、2、4h,人工肠液作用0、2、4、6、8、10h环境下,分别测定伯顿毕赤酵母菌的活菌数。结果耐酸实验中,各酸性实验组活菌数与对照组比较均下降明显(P0.01),酸度越高,活菌数越少;耐胆盐实验中,胆盐浓度越高,活菌含量越少,其中0.2%、0.3%和0.4%的胆盐浓度与对照组比较活菌数下降明显(P0.01);人工胃液实验中,经过pH 1.5人工胃液处理4h,菌株存活率达42.78%,而经过pH 2.5和3.5的人工胃液处理4h,存活率分别为138.17%和182.90%;人工肠液实验中,随着处理时间的延长,活菌数也增加,且不同时间段的活菌数差异有统计学意义(P0.05)。结论伯顿毕赤酵母菌能够在极高的酸性和较高的胆盐环境以及人工胃肠液环境中存活和定植,符合制备益生菌制剂的条件。     

不同类型益生菌制剂在胃肠道中耐受程度的对比

目的对市场销售的几种不同类型益生菌制剂进行胃、肠液耐受程度比较,评价各益生菌制剂活菌数量,为临床选择益生菌制剂提供参考。方法选择7种菌粉及某颗粒益生菌、国产某晶球、日本产某晶球、某胶囊益生菌、圣邦步步佳肠释益生菌共12种口服益生菌产品,在崩解时限仪中加入人工胃液进行崩解试验,在0.0 h、0.5 h、1.0 h、1.5 h和2.0 h时吸取上述样品并进行浓度梯度稀释,并在MRS固体培养基上培养后进行活菌计数。在上述5个时间点吸取部分益生菌-胃液混合物(针对益生菌粉末)或取出益生菌产品(针对晶球、肠溶胶囊、肠释益生菌)换人工肠液再进行40 min崩解,在MRS固体培养基上培养后进行活菌计数。结果粉末状益生菌制剂对人工胃液的耐受能力普遍较差。某颗粒益生菌、国产某晶球、日本产某晶球、某胶囊益生菌、圣邦步步佳肠释益生菌在人工胃液中不崩解。圣邦步步佳肠释益生菌在人工肠液中40 min完全崩解且活菌数峰值较高。结论圣邦步步佳肠释益生菌对人工胃、肠液耐受能力较强,且活菌数较高。     

重组干酪乳杆菌在模拟消化环境中生存性能的研究

目的 探讨重组干酪乳杆菌Lactobacillus casei 393在模拟胃肠道环境中的存活能力.方法 人工模拟胃肠道环境,即人工胃液（pH=1.5～4.5）、人工肠液、胆汁（质量浓度0.3～3.0 g/L）和高盐（质量浓度40～90g/L）.结果 重组干酪乳杆菌在pH为2.5～4.5的人工胃液中具有较强的生存能力,3 h活菌数仍达108/ml;在人工肠液中经过不同时间的作用后,重组干酪乳杆菌显出生长趋势;在0.3%的胆汁环境作用8 h仍有存活,且能耐受7%NaCl浓度的高渗环境.结论 实验为干酪乳杆菌Lactobacillus casei 393能否作为益生菌制剂在胃肠道中发挥作用提供了理论基础.     

一株广谱抑菌活性乳酸菌的筛选及特性研究

【目的】从贵州剑河采集的传统自然发酵豆酱中分离筛选具有广谱抑菌效果的乳酸菌,并进行肠道益生特性的研究。【方法】通过抑菌试验分离筛选得到菌株DJ-04,对其进行人工胃肠液耐受性、胆盐耐受性和渗透压耐受性的研究,并对其进行生理生化鉴定和16S r RNA鉴定。【结果】菌株DJ-04对大肠杆菌、沙门氏菌、金黄色葡萄球菌、志贺氏菌和铜绿假单胞菌的生长有很好的抑制作用;在p H值为2.5的人工胃液中处理3 h活菌数达到107 CFU/m L以上;在人工肠液中处理3 h活菌数达到108 CFU/m L以上,对人工胃肠液表现出良好的耐受性。能耐受一定浓度的牛胆盐,在质量浓度0.2 g/100 m L的牛胆盐环境中活菌数可达到107 CFU/m L;具有较高的渗透压耐受能力,在Na Cl质量浓度为10 g/100 m L的液体MRS中培养24 h后,活菌数仍在107 CFU/m L以上。经鉴定,DJ-04为植物乳杆菌。【结论】植物乳杆菌DJ-04具有良好的人工胃肠液耐受性以及耐胆盐和耐渗透压能力,具有肠道益生菌的潜能。     

几株饲用益生菌对体外模拟胃肠道环境的抗逆性评估

目的研究几株益生菌胃肠道环境下的抗逆能力。方法体外模拟正常猪的胃肠道环境,配制人工肠液和人工胃液,将实验室几株饲用益生菌在人工胃液和人工肠液中分别作用4、6 h,每2 h测一次活菌量。结果实验菌株对模拟胃液的耐受性都较强;除乳酸菌A外,其他菌株对模拟肠液的耐受性也较强。结论除乳酸菌A外的几株实验菌种作为饲用益生菌在抵御猪胃肠道的不良环境方面有很大优势。     

复合益生菌制剂"海生元"制备各阶段效力稳定性观察

目的 :检测复合益生菌制剂“海生元”制备储存各阶段益生菌的存活数量及其生物表型的稳定性 ,以判定该制剂的效力稳定性。方法 :在有效期内 (1年 ) ,每月定期随机取 5个“海生元”胶囊 ,按常规对M9(乳杆菌QJ4 0 5 )和T9(双歧杆菌QJ4 0 5 )两株益生菌进行活菌培养计数和菌种鉴定。结果 :(1)在常温保存条件下的一年效期内 ,制剂中的 2种益生菌 (M9和T9)存活数量略有下降趋势 ,但各次结果均符合产品的效力要求 (活菌数 >10 8/ g)。 (2 )M9和T9各次纯培养的形态学、生化反应模式及终末代谢产物气相色谱定量分析结果均分别符合其乳杆菌和双歧杆菌的菌种鉴定特征。结论 :复合益生菌制剂“海生元”在常温保存的 1年有效期内 ,可保持制剂中益生菌的活菌数量及稳定的生物表型特征 ,从而保证该制剂的效力稳定性和疗效要求。     

新疆传统发酵乳品中乳酸菌与 酵母菌的益生特性

目的观察新疆传统发酵乳品中分离的14种菌株的生长特点及产酸能力,筛选出具有较强耐胆盐能力,并能在人工胃肠液中存活的菌株。方法对10株乳酸菌和4株酵母菌进行生长曲线、pH、耐胆盐能力和耐人工胃肠液检测。结果 10株乳酸菌和4株酵母菌具有良好的生长曲线和产酸能力;马乳酒样乳杆菌具有较强的耐胆盐能力;希氏乳杆菌、马乳酒样乳杆菌、乙醇假丝酵母和东方伊萨酵母具有较强的耐人工胃液能力;乳酸乳球菌、哈尔滨乳杆菌、瑞士乳杆菌、马乳酒样乳杆菌、乙醇假丝酵母和东方伊萨酵母具有较强的耐人工肠液能力。结论 10株乳酸菌和4株酵母菌具有优良的益生特性,有望成为益生菌制剂的备用菌株。     

栀子豉汤所含优势益生菌群的研究

目的考察不同煎煮方法及人工胃液、肠液对栀子豉汤所含优势菌群的影响,基于芽胞杆菌的整肠作用探讨栀子豉汤的作用机制。方法采用先煎栀子再煎淡豆豉(方法一)及两药同时煎煮(方法二)得到不同的栀子豉汤溶液,在营养琼脂培养基上培养及菌落计数,通过Biolog自动微生物鉴定系统对分离菌株进行鉴定,并考察人工胃液、肠液对栀子豉汤所含优势菌群数量的影响。结果方法一制备的栀子豉汤所含优势菌数量更多,为(2.55±0.02)×10~5 CFU/mL,以枯草芽胞杆菌为主;栀子豉汤经人工胃液、人工肠液作用后,优势菌又大量繁殖。结论栀子豉汤可能通过其所含益生芽胞杆菌的整肠作用而发挥药效。     

HRP在模拟人体胃肠环境中的稳定性研究

以中国药典中的人工胃液和人工肠液模拟人体胃肠环境,对过辣根氧化物酶的稳定性进行了一系列单独及综合研究。结果表明,辣根过氧化物酶在人工胃液中受到很强的破坏,20min后,其活性仅存1.1%,主要由胃酸破坏,而在人工肠液中则较为稳定,1.5h后,其活性仍存在89%,同时还发现,天然结构的辣根过氧化物酶对胰蛋白酶具有较强的耐受力。     

静电喷雾法制备乳酸菌微胶囊的实验研究

目的为解决乳酸菌产品活菌数的不稳定性，对乳酸菌进行微胶囊化包埋。方法用海藻酸钠和明胶的混合体系作为壁材，对乳酸菌进行静电喷雾包埋处理，并让微胶囊化乳酸菌在模拟胃肠液的环境中进行耐酸性和肠溶性实验。结果混合体系的壁材与乳酸菌具有较好的生物相容性，优选得出当芯壁材为12时包埋率最高（96．3％），微胶囊化乳酸菌在经人工胃液处理2h后，活菌数比未经微胶囊化的对照组高出2个数量级，且在经人工肠液处理40min后，乳酸菌几乎全部释放。结论静电喷雾法制备的乳酸菌微胶囊具有一定耐酸性和肠溶性。     

罗汉果皂苷V在人工胃液中的稳定性和体外代谢研究

为了研究罗汉果皂苷V在胃肠道中的稳定性及体外代谢特征,该研究首先考察罗汉果皂苷V在人工胃液中的稳定性,然后在体外模拟人肠内环境,研究人肠内细菌对该成分的代谢作用,并采用LCMS/IT-TOF液质联用仪分析了培养液中的化学成分,考察了罗汉果皂苷V在人工胃液和人肠道菌群培养液中的变化。结果表明:罗汉果皂苷V在人工胃液条件下,迅速发生酸水解反应,分别脱去3个糖苷、4个糖苷、5个糖苷及2个氢转化为二糖苷、一糖苷与脱氢苷元,其脱糖反应随着时间的延长而反应越完全,反应4 h后就只剩下苷元; 该化合物在人的肠道菌群作用下经过脱糖反应转化为四糖苷、三糖苷、二糖苷、一糖苷,经过糖苷化反应转化成六糖苷。通过体外实验,初步了解罗汉果皂苷V在胃液和肠道环境中的降解规律,该研究结果为今后探讨罗汉果皂苷在人体内的代谢与生物转化规律提供了物质和技术基础。     

Influence of encapsulation on the survival of probiotics in food matrix under simulated stress conditions

The main objective of the present study was to evaluate the influence of encapsulation by extrusion technique using two hydrogels, namely; sodium alginate (Na-ALG) and whey protein isolate (WPI) on Bifidobacterium bifidium viability and stability of yoghurt under simulated gastrointestinal conditions. Probiotic bacteria (free or encapsulated) were added to yogurt for four weeks to test their viability and stability. Physicochemical and sensory analysis of yoghurt were conducted. Viability of B. bifidium in the simulated gastrointestinal conditions pH 2 and pH 7.5 was determined. Also, the efficiency of encapsulated final yield of the microcapsules was determined. With storage time, the pH of yoghurt containing encapsulated bacteria increased more than that of yoghurt containing free probiotic bacteria, resulting in a decrease in acidity. When compared to yoghurt containing encapsulated bacteria, the lactose level of yoghurt containing free probiotic bacteria decreased over time. The viscosity of yoghurt containing encapsulated WPI remained stable over the storage period, with syneresis remaining stable. The sensory properties of yoghurt containing free probiotics deteriorated over time. Cell viability was significantly reduced in yoghurt-containing free probiotics compared to other treated yoghurts. Cell viability in free probiotics yoghurt was lower than in encapsulated ones when exposed to simulated gastric and intestinal juice. In conclusion, WPI- encapsulated probiotics showed better stability over 28 days of storage in both yoghurt and gastrointestinal conditions, followed by sodium alginate.     

Isolation and characterization of anti-Salmonella lactic acid bacteria from the porcine gastrointestinal tract

AIMS: To identify lactic acid bacteria (LAB) of porcine intestinal origin with anti-Salmonella activity. METHODS AND RESULTS: Samples were obtained from pig faeces and caeca and screened for the presence of anti-Salmonella LAB. The 11 most promising isolates were identified as belonging to the genera Lactobacillus and Pediococcus. The LAB exhibited large variation in their ability to survive in simulated gastric juice at pH 1.85. While Lactobacillus johnsonii species survived at levels of 80% for up to 30 min, Lactobacillus pentosus species declined to <0.001% in that time. All isolates tolerated porcine bile at a concentration of 0.3% (w/v), with some isolates capable of growth in the presence of up to 5% (w/v) bile. The ability of the LAB isolates to prevent Salmonella invasion of intestinal epithelial HT-29 cells varied, with reductions of between 30% (Lact. pentosus) and 80% (Lactobacillus murinus spp.) observed. CONCLUSIONS: LAB of porcine origin were observed to survive simulated passage through the GIT and inhibit growth of Salmonella and its invasion of the intestinal epithelium. SIGNIFICANCE AND IMPACT OF THE STUDY: The data demonstrate that some porcine intestinal LAB isolates may offer potential as probiotics for the reduction of Salmonella carriage in pigs.     

利用培养组学技术分离鉴定体外降解胆固醇肠道细菌及其功能评价

高胆固醇是诱发心脑血管疾病的重要因素之一。目前国内外降低胆固醇的主要方式是药物治疗,但其存在费用高及副作用多的弊端。研究表明,肠道细菌对机体的胆固醇代谢有重要影响,但降胆固醇肠道细菌的筛选方式及功能评价却少有报道。本研究通过培养组学方法,使用牛胆汁酸或人工混合胆汁酸作为筛选条件,从健康人体肠道筛选出36种耐胆汁酸细菌。以鼠李糖乳杆菌GG株 (Lactobacillus rhamnosus GG,LGG) 作为阳性对照,设置0 g/L、0.3 g/L、3 g/L三种胆汁酸浓度组,对耐胆汁酸细菌进行体外降胆固醇能力评估,确定奇异变形菌、斯氏普罗威登斯菌、普通变形菌等10种细菌为降胆固醇优势菌。随后对其中6种降胆固醇优势菌——奇异变形菌、斯氏普罗威登斯菌、普通变形菌、潘氏变形菌、污蝇解克杆菌、雷氏普罗威登斯菌进行体外降甘油三酯能力评估以及人工胃液耐受能力评估。结果显示,上述6株细菌体外降甘油三酯能力均优于LGG。伴随人工胃液pH值的下降和作用时间的延长,6株细菌的生存率均下降。上述筛选实验及功能评价为进一步开发潜在的降胆固醇细菌制品提供研究基础。     

模拟胃肠环境对3株光合细菌存活率影响的实验研究

目的模拟胃肠环境对3株光合细菌(Photosynthetic bacteria,PSB)存活率影响进行体外实验研究。方法测定3株PSB分别在pH2.0、3.0和4.0的模拟胃液及含有或不含有0.45%牛胆盐(pH8.0)的模拟小肠液中不同时间的存活率。结果pH2.0时模拟胃液处理不同时间对3株PSB存活率有显著影响(P0.01);pH3.0时,模拟胃液处理60、90和180 min对菌株rub存活率有显著影响(P0.01),处理90、180 min对菌株Ga存活率有显著影响(P0.05),处理180 min对菌株2C存活率无显著影响(P0.05);pH4.0时模拟胃液处理180 min对3株PSB存活率无显著影响(P0.05)。模拟小肠液处理240 min对3株PSB菌株活菌数无显著影响(P0.05)。结论模拟胃液对3株PSB存活率影响具有菌株特异性和pH依赖性,而模拟小肠液对3株PSB存活率无显著影响。     

Lactobacillus cells in the rabbit digestive tract and the factors affecting their distribution

Lactobacillus fermentum was present in small numbers in the caecum and colon of only one of 16 rabbits. Studies aimed at elucidating the factors affecting the colonization of lactobacilli in rabbit gut were thus performed. These studies included investigation of the effect of gastric juice and bile salts on the viability of lactobacilli, assay of the survival rates of lactobacilli in the gastrointestinal tracts of the rabbits with or without ileum cannulation and measurement of the adhesive capability of lactobacilli to the rabbit intestinal epithelial cells. Results showed that, although some lactobacilli were resistant to the rather low pH levels of rabbit gastric juice, lack of adhesive capability may prevent them from colonizing in the intestinal tract. Therefore, if lactobacilli are to be used as probiotics for rabbit, factors that affect their viability and ability to colonize should be considered.     

Transformation of ethyl alcohol to ethyl nitrite in acidified saliva: possibility of its occurrence in the stomach

After taking alcoholic beverages, the ethanol is mixed with saliva and then gastric juice. As pH of gastric juice is around 2, the ethanol might be transformed to ethyl nitrite in the stomach by reacting with salivary nitrite. In this study, reactions between ethanol and nitrite in acidified saliva were investigated. The result indicates that nitrite in acidified saliva reacted with ethanol producing ethyl nitrite. It is discussed that ethyl nitrite might be formed in the stomach after drinking alcoholic beverages and that the ethyl nitrite might function as a donor of NO in intestinal and gastric tissues.     

蒙古族儿童源益生特性双歧杆菌的筛选及鉴定

【目的】双歧杆菌在人和动物胃肠道中发挥着重要的生理作用,然而双歧杆菌能否耐受胃酸、肠液及胆汁酸是影响活菌制剂益生效果的关键因素,本研究旨在从蒙古族儿童粪便中分离筛选出具有良好益生特性的双岐杆菌。【方法】本文采用双岐杆菌选择性培养基对样品进行分离纯化,并对菌株进行生理生化鉴定,以耐受人工胃肠液、耐受牛胆盐为手段对各菌株的益生特性进行评价,并且对B. animalisV9进行了16S rDNA分子生物学鉴定。【结果】本文从12位健康蒙古族儿童粪便中分离得到11株双歧杆菌,经传统生理生化试验鉴定为5株B. adolescentis：A1、H3、G4、A8、V10;3株B. longum：C6、C7、D11;1株B. pseudocatenlatum：B2;1株B. bifidum：G5;1株B. animalis：V9。B. animalis V9具有较强的耐酸性,在pH2.0的人工胃液中厌氧培养3h后存活率为92.4%,而其它10株双歧杆菌在此条件下的存活率均小于31.25%;B. animalis V9在pH2.0的人工胃液中厌氧培养3h后接入pH8.0的人工肠液中消化8h,存活率为99.7%,并且可以耐受0.3%的牛胆盐。进一步对V9菌株进行16S rDNA分子生物学鉴定,发现与Bifidobacterium animalis subsp. lactic BB12的同源性为99%。【结论】本研究结果显示B. animalis V9来源安全,并且具有良好耐酸、耐胆盐益生特性,有望在乳制品及保健类产品中得到广泛的应用。