X-Gal在双歧杆菌分离与活菌计数方面的应用研究

利用大多教双歧杆菌具有半孔糖苷酶且活性较高的特性,在NPNL培养基中加入X-Cal(5-溴-4-氯-3-吲哚-β-D-半乳糖苷)作为底物,双歧杆菌水解底物释放出吲哚,产生颜色反应。双歧杆菌菌落呈深蓝色,乳酸菌和其它细菌一般为白色或淡蓝色。将X-Gal应用于不同样品中双歧杆菌分离和活菌计数进行对比。这种方法能比较准确的反映双歧制品和婴儿粪便中双歧杆菌的数量,并且大大减少了分离鉴别双歧杆菌的工作量。     

一种发酵乳中双歧杆菌鉴别培养基计数评估

目的:建立发酵乳中双歧杆菌数量计数测定方法。方法:采用果糖为碳源的鉴别培养基,基于乳酸菌同型发酵和异型菌落形态的差异,借助pH指示剂确定样品中双歧杆菌数量。结果:该培养基所计双歧杆菌数量与对照培养基所计数量在同一数量级且差异不显著(P>0.05)。结论:该培养基可用于发酵乳中双歧杆菌的准确计数。     

功能性酸乳中每种益生菌的单独计数方法及其混合发酵工艺的研究

通过研究分析功能性酸乳中双歧杆菌和各种乳酸菌在BBL平板培养基上的菌落特征及菌体形态,提出了一种可同时准确对每种益生菌单独计数的新方法,从而可直观地观察到各种益生菌的生长活度,免除了传统方法计数不同有益菌数要用不同培养基的繁琐,为生产和检测提供了依据;还对功能性酸乳的混合发酵生产工艺进行了研究,确定了最佳的菌种比例,按嗜热链球菌3%、保加利亚乳杆菌2%、嗜酸乳杆菌4%、双歧杆菌10%接种,39℃发酵11~13h,酸乳产品既保持了普通酸乳的良好口感和风味,又保证了双歧杆菌和乳酸菌活菌数在贮存期内均大于108cfu/mL,增强了酸乳的保健功能。     

选择性计数双歧杆菌培养基的效果比较

随着双歧杆菌基础研究和开发应用的迅速发展,双歧杆菌活菌计数方法越来越重要,实验目的是要选择1种适于选择性计数产品中双歧杆菌菌落数的培养基。主要采用菌落技术方法,比较了常用的8种选择性培养基对7种产品中双歧杆菌的选择性计数效果。结果表明,以改良MRS-X-Gal琼脂培养基上形成的双歧杆菌菌落数最高,且菌落特征明显,易于计数,是优选出的效果较好的选择性计数培养基。     

乳酸菌菌粉定量计数方法研究

目的:以9株乳杆菌、6株双歧杆菌、3株球菌、1株凝结芽孢杆菌菌粉为研究对象,在国标GB 4789.35-2016的基础上,对稀释倍数、稀释液成分、培养基成分进行比较研究,考察对乳酸菌菌粉计数活菌数的影响。方法:采用稀释平板计数的方法,对不同的乳酸菌菌粉进行活菌计数。结果:初始乳酸菌菌粉样品稀释倍数对最终计数活菌数无明显影响,ISO稀释液对部分乳杆菌、双歧杆菌菌粉的活菌计数结果有显著提高(P<0.05);双歧杆菌菌粉采用TOS琼脂培养基的计数结果显著优于国标培养基(P<0.05);凝结芽孢杆菌菌粉采用改良芽孢计数培养基计数结果优于PCA和NA计数培养基。结论:平板计数方法中,稀释液中含有酪蛋白胨能提升双歧杆菌菌粉的活菌计数数量;TOS琼脂培养基更有利于双歧杆菌的增殖培养;改良芽孢计数培养基更有利于凝结芽孢杆菌的芽孢萌发增殖。     

X-Gal在双歧杆菌分离与活菌计数方面的应用研究

利用大多数双歧杆菌具有半乳糖苷酶且活性较高的特性，在NPNL培养基中加入X—Gal(5-溴-4-氯-3-吲哚-8-D-半乳糖苷)作为底物，双歧杆菌水解底物释放出吲哚，产生颜色反应。双歧杆菌菌落呈深蓝色，乳酸菌和其它细菌一般为白色或淡蓝色。将X—Gal应用于不同样品中双歧杆菌分离和活菌计数进行对比。这种方法能比较准确的反映双歧制品和婴儿粪便中双歧杆菌的数量，并且大大减少了分离鉴别双歧杆菌的工作量。     

乳制品中选择性分离与计数植物乳杆菌的方法

根据乳酸菌对抗生素的敏感性、产酸能力和碳源利用不同,研制出植物乳杆菌分离计数培养基(LPMRS培养基),通过对9种不同的乳酸菌菌株单独试验及混合试验,对该培养基的特异性进行了评价。通过对LP-MRS培养基和MRS培养基中植物乳杆菌计数比较,对该培养基是否抑制植物乳杆菌生长进行了验证。结果表明,该培养基可抑制乳酸乳球菌乳酸亚种、嗜热链球菌、瑞氏乳杆菌、嗜酸链球菌、婴儿双歧杆菌的生长,并能将植物乳杆菌从干酪乳杆菌群分离开来,且对植物乳杆菌无抑制作用。该方法操作简便,成本低,可广泛应用于乳制品中植物乳杆菌的分离和计数。     

益生菌产品中双歧杆菌计数培养基的比较研究

对添加单株双歧杆菌(分别为乳双歧杆菌BB12和长双歧杆菌BB536)的益生菌产品进行双歧杆菌计数实验.比较了在相同的稀释条件和培养条件下,倾注MRS、半胱氨酸盐酸盐+MRS(CYS-MRS)、BBL3种培养基,培养计数结果的差异.结果对添加乳双歧杆菌BB12的益生菌产品,3种培养基计数结果差异较显著(P＜0.05),B...     

发酵乳中乳酸菌的选择性计数及分离鉴定

发酵乳产品的菌种组成与活菌数的含量是产品质量的关键。该文通过选择性培养基、基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry, MALDI-TOF-MS)快速鉴定技术及多相鉴定技术探究3种发酵乳中乳酸菌的活菌数及种类。研究结果表明,3种发酵乳产品中,GB 4789.35对乳酸菌总数的计数结果均高于选择性培养基,M17培养基对嗜热链球菌的计数结果均高MC培养基。MALDI-TOF MS快速鉴定表明M17、MRS(含50μg/mL的万古霉素)、双歧杆菌培养基可以选择性的分离发酵乳中的嗜热链球菌(Streptococcus thermophilus)、干酪乳酪杆菌(Lacticaseibacillus casei)和双歧杆菌(Bifidobacterium spp.),酸化MRS培养基对德氏乳杆菌(Lactobacillus delbrueckii)的选择性较差,不能在含有双歧杆菌的发酵乳中选择性分离德氏乳杆菌。进一步通过多相鉴定确定3种发酵乳产品中乳酸菌在...     

嗜酸乳杆菌、双歧杆菌和干酪乳杆菌的选择性计数

介绍了干酪乳杆菌、嗜酸乳杆菌和双歧杆菌的选择性计数方法。LC培养基只能计数干酪乳杆菌,MRS-水杨素（或山梨醇）培养基可以计数嗜酸乳杆菌和干酪乳杆菌;而MRS培养基可以计数这3种益生菌,通过减法原则从嗜酸乳杆菌、干酪乳杆菌和双歧杆菌混合物中单独计数。另外,MRS-NNLP培养基也可用于选择性计数双歧杆菌。     

双歧酸乳最佳发酵条件的研究

双歧杆菌作为一种益生菌,其促进人体健康的有益作用越来越受到人们的重视,双歧杆菌乳制品利用双歧杆菌的益生作用在功能性乳制品的研发过程中也占有重要的一席之地.该文主要研究双歧杆菌酸乳的发酵条件,利用正交试验优化双歧杆菌与乳酸菌混合发酵的工艺条件,确定双歧杆菌和乳酸菌在牛乳中混合发酵的适宜接种量4％、发酵温度40℃、菌种配比(保加利亚乳杆菌∶嗜热链球菌∶双歧杆菌)3∶2∶1.     

不同乳酸菌在冬瓜汁中的发酵特性研究

向鲜榨并经巴氏灭菌的冬瓜汁中接入不同的乳酸菌(干酪乳杆菌、嗜热链球菌、植物乳杆菌、嗜酸乳杆菌、鼠李糖乳杆菌、双歧杆菌和保加利亚乳杆菌)进行发酵,比较发酵过程中乳酸菌活菌数、pH值、可滴定酸、总糖、还原糖、总多酚和色泽等变化和发酵后挥发性风味物质种类与相对含量。结果表明,冬瓜汁营养丰富,上述各种乳酸菌均能在冬瓜汁中很好地生长,其中干酪乳杆菌的生长速率(对数生长期)略低于其他6种菌;另外,七种乳酸菌发酵后冬瓜汁中主要挥发性风味物质主要有酸类、醇类、酯类、醛类、酮类五类,保加利亚乳杆菌发酵的冬瓜汁中检出的挥发性风味物质最多。除双歧杆菌之外其他六种乳酸菌发酵的冬瓜汁理化性质差别不大,双歧杆菌发酵的冬瓜汁在总色差上明显高于其他乳酸菌,且双歧杆菌的产酸能力最强,在发酵期间其可滴定酸含量最高。     

乳酸菌及双歧杆菌制品的生产工艺

介绍了乳酸杆菌和双歧杆菌生物学特性,及它们对人体健康的功能及相应保健食品的生产工艺.     

定量PCR技术检测杀菌型产品中乳酸菌

目的利用定量PCR(quantitativePCR,qPCR)技术检测杀菌型产品中乳酸菌,并与标签标示相比较,查探产品中各类菌种的异同性及数量。方法以市售的发酵乳及含乳饮料作为研究对象,设计可区分产品中常见的6种乳酸菌特异性引物及探针,利用qPCR技术对方法的特异性、灵敏度进行验证,并建立标准扩增曲线;利用模拟样品对方法进行检验,最后对市场上购买的实际样品进行检测。结果除去干酪乳杆菌,其他5种引物都能特异性扩增出其目标菌,并且对其他的乳酸菌均无扩增现象;DNA检测的灵敏度可达到2～4pg;单一标准菌种扩增曲线线性较好,相关系数r2值在0.954～0.992之间;对模拟样品的检测,与平板法差异性比较,P值均大于0.05,无显著性差异。对市售样品进行检测,产品中标记的干酪乳杆菌、双歧杆菌和嗜热链球菌与检测结果存在不匹配现象。结论本研究建立的qPCR方法可快速、准确地对产品中德氏乳杆菌保加利亚亚种、嗜热链球菌、乳酸乳球菌、嗜酸乳杆菌、双歧杆菌的鉴定和定量的分析。     

Simplified Direct Plating Method for Enhanced Recovery of Escherichia coli in Food

A refined plating medium designed to enumerate E. coli in food within 24 hr was developed. The medium combined tergitol 7 with the fluorogenic substrate, 4-methylumbelliferyl β-D-glucuronide. E. coli colonies were detected by fluorescence under long-wave UV light. The new agar allowed superior recovery of heat and freeze-stressed cells and produced E. coli counts from meat and poultry samples at levels generally equivalent to or better than those of the standard 5 tube Most Probable Number method. This method provides a simple and rapid alternative to traditional techniques.     

Selective enumeration of Lactobacillus delbrueckii ssp. bulgaricus,Streptococcus thermophilus,Lactobacillus acidophilus,bifidobacteria, Lactobacillus casei,Lactobacillus rhamnosus,and propionibacteria

Nineteen bacteriological media were evaluated to assess their suitability to selectively enumerate Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus, bifidobacteria, and propionibacteria. Bacteriological media evaluated included Streptococcus thermophilus agar, pH modified MRS agar, MRS-vancomycine agar, MRS-bile agar, MRS-NaCl agar, MRS-lithium chloride agar, MRS-NNLP (nalidixic acid, neomycin sulfate, lithium chloride and paramomycine sulfate) agar, reinforced clostridial agar, sugar-based (such as maltose, galactose, sorbitol, manitol, esculin) media, sodium lactate agar, arabinose agar, raffinose agar, xylose agar, and L. casei agar. Incubations were carried out under aerobic and anaerobic conditions at 27, 30, 37, 43, and 45 degrees C for 24, 72 h, and 7 to 9 d. S. thermophilus agar and aerobic incubation at 37 degrees C for 24 h were suitable for S. thermophilus. L. delbrueckii ssp. bulgaricus could be enumerated using MRS agar (pH 4.58 or pH 5.20) and under anaerobic incubation at 45 degrees C for 72 h. MRS-vancomycine agar and anaerobic incubation at 43 degrees C for 72 h were suitable to enumerate L. rhamnosus. MRS-vancomycine agar and anaerobic incubation at 37 degrees C for 72 h were selective for L. casei. To estimate the counts of L. casei by subtraction method, counts of L. rhamnosus on MRS-vancomycine agar at 43 degrees C for 72 h under anaerobic incubation could be subtracted from total counts of L. casei and L. rhamnosus enumerated on MRS-vancomycine agar at 37 degrees C for 72 h under anaerobic incubation. L. acidophilus could be enumerated using MRS-agar at 43 degrees C for 72 h or Basal agar-maltose agar at 43 degrees C for 72 h or BA-sorbitol agar at 37 degrees C for 72 h, under anaerobic incubation. Bifidobacteria could be enumerated on MRS-NNLP agar under anaerobic incubation at 37 degrees C for 72 h. Propionibacteria could be enumerated on sodium lactate agar under anaerobic incubation at 30 degrees C for 7 to 9 d. A subtraction method was most suitable for counting propionibacteria in the presence of other lactic acid bacteria from a product. For this method, counts of lactic bacteria at d 3 on sodium lactate agar under anaerobic incubation at 30 degrees C were subtracted from counts at d 7 of lactic bacteria and propionibacteria.     

原生物酸奶及其发展前景

介绍了原生物、乳酸菌、双歧因子的特点和种类，原生物酸奶的市场情况、菌种、生产、保健作用及发展趋势。原生物酸奶利用最多的是嗜酸乳杆菌和双歧杆菌，二者在抵抗上消化道酸性环境方面有协同作用。生产时需与传统菌种配合，发酵时间约６ｈ。酸奶中原生物可采用一种平板培养法单独计数。改良菌种和添加双歧因子，有望提高酸奶的保健效果。     

Comparison of dichloran 18% glycerol (DG18) agar with general purpose mycological media for enumerating food spoilage yeasts

Dichloran 18% glycerol (DG18) agar was originally developed to enumerate xerophilic foodborne moulds. However, some laboratories are using DG18 agar as a general medium to enumerate foodborne moulds and yeasts. A collaborative study, with the participation of seven laboratories, was undertaken to compare DG18 agar with dichloran rose bengal chloramphenicol (DRBC) agar, tryptone glucose yeast extract chloramphenicol (TGYC) agar, and plate count agar supplemented with chloramphenicol (PCAC) for enumerating 14 species of common food spoilage yeasts. Comparison of the mean values of populations of all yeasts recovered on each medium revealed no significant differences among DRBC agar, PCAC, and TGYC agar, while each of these media supported the development of significantly (P < or = 0.05) higher numbers of colonies than DG18 agar. However, differences were only 0.08 to 0.10 log10 cfu/ml, making the practical significance questionable. The overall coefficient of variation (CV) for within laboratory repeatability was 1.71%, while the CV for reproducibility of counts obtained among laboratories was 6.96%. Compared to DRBC agar, TGYC agar, and PCAC, yeast colonies were smaller on DG18 agar. Growth of Brettanomyces anomalus, Cryptococcus albidus, and Rhodotorula mucilaginosa was particularly retarded or inhibited on DG18 agar. Based on the performance of media in supporting colony development and ease of counting colonies, the use of DG18 agar as a general enumeration medium for foodborne yeasts cannot be recommended.     

酸乳储藏中双歧杆菌活性影响因素研究

以改良MRS液体培养基为保存基质,以双歧杆菌活菌数为考查指标,针对酸乳特性研究了基质起始pH、冷藏温度及混合菌种3因素对双歧杆菌活性的影响。结果表明:(1)不同的基质起始pH值对双歧杆菌存活的影响极显著,基质起始pH5.5时对双歧杆菌活性的影响最小,并且起始pH值提高到5.5可使双歧杆菌保持最小保健剂量(106cfu/mL)的时间延长至20d。(2)当基质起始pH为4.6时,双歧杆菌在4℃保存与在室温(25℃)保存活性情况基本一致。(3)双歧杆菌无论以单一菌种形式保存还是与嗜酸乳杆菌以1:1的比例混合保存,其活性情况无明显变化,即嗜酸乳杆菌对双歧杆菌无明显的促进或抑制作用。