混合制剂中快速计数双歧杆菌鉴别培养基的研究

研制了一种混合制剂中快速计数双歧杆菌鉴别培养基。用添加X-Gal的改良MRS培养基和厌氧胶法对4种标准双歧杆菌(长双歧杆菌、短双歧杆菌、青春双歧杆菌、两歧双歧杆菌)和2种标准乳酸菌(保加利亚乳杆菌、嗜热链球菌)分别单独培养、混合培养,结果在X-Gal培养基上,双歧杆菌单独培养及与乳酸菌混合培养双歧杆菌均为白色菌落或浅蓝色菌落,而乳酸菌为蓝色菌落。结果表明,在X-Gal培养基上双歧杆菌和乳酸菌有明显的鉴别性,可用于双歧杆菌和乳酸菌混合制剂的菌落鉴别计数。     

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。     

婴幼儿乳粉内双歧杆菌简便计数法的研究北大核心CSCD

选择适用于婴幼儿配方粉的3株双歧杆菌作为研究菌株,通过设计不同的平板类型和培养方式来验证适用于以上3株双歧杆菌的最优计数方式。结果:双层平板(下层为20 mL MRS,上层为10 mL石蜡)可以无需厌氧培养箱即可实现对含动物双歧杆菌Bb-12、乳双歧杆菌Bi-07或乳双歧杆菌HN019产品的可靠计数,而且检测结果与厌氧培养箱法无显著差异。     

双层包埋型益生菌制品检测方法的研究

为了研究双层包埋型益生菌制品的检测方法,采用生理盐水和专用稀释液分别对双层包埋型益生菌制品进行样品前处理后,使用改良MC琼脂、MRS琼脂培养法进行乳酸杆菌计数培养的比较,同时采用TPY琼脂、BL琼脂和BBL琼脂培养基,使用厌氧培养法,进行双歧杆菌计数培养的比较.经专用稀释液处理的制品其益生菌数量检出率较采用生理盐水处理的高10倍,且乳酸杆菌的检测以MRS培养基为佳,双歧杆菌的检测以BBL培养基为佳.可见使用专用稀释液是进行双层包埋型益生菌制品样品前处理的较好方法,可以显著提高制品的乳酸杆菌和双歧杆菌的检出率.     

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。     

双歧杆菌在发酵乳中活性的研究

分别使用两歧双歧杆菌和长双岐杆菌制作酸乳并冷藏21d,应用AMC,RMS培养基检测样品的双歧杆菌活菌数。发酵乳中的双歧杆菌的浓度达到10^8-10^9CFU/mL,在21d的冷藏中二种双歧杆菌的活菌数都超过了最小保健剂量10^6CFU/mL。RMS培养基同样有效地分离两歧双歧双歧杆菌和长双歧杆菌,可以采用涂布法和倾注法进行操作,RMS是对酸乳中双歧杆菌计数的最适宜的选择性培养基,使用AMC培养基由于涂布法造成较低的活菌计数。     

TTC在乳及乳制品菌落总数检测中的应用

检测乳及乳制品菌落总数,为达到菌落在培养基中清晰可辨,在平板计数琼脂培养基中添加2,3,5-三苯基氯化四氮唑(TTC)。以灭菌乳、发酵乳、冰淇淋为样品基质,分别加入适当菌含量的大肠埃希氏菌、金黄色葡萄球菌、粪肠球菌、枯草芽胞杆菌工作菌悬液,制备适当浓度的TTC平板计数琼脂,同时以不添加TTC的平板计数琼脂作为对照,对同一样品进行菌落计数并同国标方法结果进行对比。TTC平板计数琼脂终浓度在0.001%~0.002%时能够使菌落显色,且与不添加TTC的对照组对比结果一致。在检测成分复杂的乳及乳制品样品时添加适宜浓度的TTC,有助于菌落的辨认,能够提高检验人员菌落计数的准确性,可用于检测乳及乳制品中菌落总数。     

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...     

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...     

发酵奶中乳酸菌菌种检出及活菌计数调查

目的分析北京市场发酵奶(酸奶)在保质期内的乳酸菌数及乳酸菌菌种的检出率。方法对11个酸奶厂家的20种不同酸奶产品进行乳酸菌活菌计数及所用菌种的检验。结果在保质期间,双歧杆菌和嗜酸乳杆菌检出率分别为23.08%和27.27%;保加利亚乳杆菌与嗜热链球菌分别为72.22%和94.44%。在保质期末,嗜热链球菌的平均活菌数为3.23×106CFU/ml,保加利亚乳杆菌为4.17×105CFU/ml,双歧杆菌为1.12×104CFU/ml,嗜酸乳杆菌为1.32×104CFU/ml。结论酸奶中嗜热链球菌和保加利亚乳杆菌的检出率及活菌数均高于双歧杆菌和嗜酸乳杆菌。     

Antimicrobial activity of neutralized extracellular culture filtrates of lactic acid bacteria isolated from a cultured Indian milk product (‘dahi’)

Neutralized extracellular culture filtrate obtained from isolates of Lactobacillus acidophilus, Lactobacillus delbruecki ssp. bulgaricus, Lactobacillus salivarius and Lactococcus lactis ssp. lactis from ‘dahi’ showed weal to moderate inhibition of Staphylococcus aureus, Bacillus cereus, Escherichia coli, Bacillus brevis, Bacillus circulans, Bacillus coagulans, Bacillus laterosporus, Bacillus subtilis and Pseudomonas aeruginosa when tested by the diffusion agar well assay method. The effective minimum quantity of lactic culture filtrates required to obtain complete inhibition of an inoculum of 10³ cfu/ml of the bacteria tested was between 20 and 26% (vol/vol), as determined by the agar incorporation method. Neutralized extracellular culture filtrate of these lactic cultures added at a level of 10% in sterile, 10% reconstituted non-fat dry milk was able to either suppress or retard growth of selected bacterial cultures when incubated at 37°C for 24 h. This study indicated the antimicrobial activity of dahi and the potential of using neutralized extracellular culture filtrate of lactic acid bacteria in the biopreservation of foods.     

Bacterial populations associated with a sorghum-based fermented weaning cereal.

Microbiological surveys, to determine the quality and safety, were conducted on 45 sorghum samples comprising dry powders (n = 15) and corresponding fermented (n = 15) and cooked fermented porridge (n = 15) samples collected from households in an informal settlement of the Gauteng Province of South Africa. Mean aerobic plate counts, Gram-negative counts and bacterial spore counts of sorghum powder samples decreased in fermented and cooked fermented porridge samples. However, mean lactic acid bacteria counts increased in fermented porridge samples, but decreased slightly in cooked fermented porridge samples. The mean pH value of sorghum powder samples decreased in fermented and cooked fermented porridge, respectively. Bacillus (B.) cereus was detected in all 15 sorghum powder samples, while Escherichia (E.) coli was detected in 53%, Clostridium perfringens in 27%, Listeria monocytogenes in 13% and Aeromonas spp., Salmonella spp., Staphylococcus aureus, Shigella spp. and Yersinia spp., each in 7% of sorghum powder samples. Of the fermented porridge samples, 40% contained B. cereus and 7% contained E. coli. None of the pathogens tested for were detected in cooked fermented porridge samples. B. cereus (53%), B. subtilis (21%), B. thuringiensis (13%), B. licheniformis (10%) and B. coagulans (3%) were identified from 120 isolates randomly selected from spore count plates of the highest dilution showing growth.     

益生菌发酵苹果汁工艺优化及有机酸的变化

以红富士苹果为原料,进行乳酸菌发酵苹果汁工艺条件优化并分析发酵期间苹果汁有机酸的变化。选择Lactobacillus paracasei 20241、Bifidobacterium animalis 6165、Streptococcus thermophilus 6063和Lactobacillus acidophilus 6005混合发酵苹果汁,以活菌数和感官评分为主要指标,在单因素试验基础上进行响应面优化试验,研究不同的菌种比例、接种量、发酵时间等对苹果汁活菌数和感官评分的影响。结果表明,乳酸菌发酵苹果汁的优化工艺条件为菌种比例1∶1∶1∶1、接种量2%、发酵时间24 h、发酵温度37℃,在此条件下得到的活菌数为1.985×108 CFU/mL,感官评分为80.23分。采用最佳工艺条件发酵苹果汁,利用高效液相色谱法对发酵过程中苹果汁有机酸的变化情况进行分析,结果表明:经过发酵后,苹果酸和琥珀酸含量下降明显(P0.05),而乳酸、奎宁酸、柠檬酸、酒石酸、丙酮酸和莽草酸含量均显著提高(P0.05)。     

益生菌发酵枣粉喷雾干燥条件的优化

以植物乳杆菌（Lactobacillus plantarum）、凝结芽孢杆菌（Bacillus coagulans）混菌对大枣进行液态发酵,通过单因素试验研究喷雾干燥进口温度、雾化流量、蠕动泵转速和脱脂乳粉添加量对益生菌发酵枣粉出粉率、感官评定、含水量、堆积密度及活菌数的影响,确定喷雾干燥进口温度、雾化流量、蠕动泵转速和脱脂乳粉添加量4个因素为自变量,感官评分为响应值进行响应面优化。结果表明,喷雾干燥制备益生菌发酵枣粉的最优参数为喷雾干燥进口温度150 ℃,雾化流量473 L/h,蠕动泵速20%,脂乳粉添加量2.5%。在此优化条件下,益生菌发酵枣粉出粉率为39.15%、感官评分为95.6分、含水量3.82%、堆积密度为0.251 g/mL、活菌数为1.78×109 CFU/g。     

藜麦酸奶混菌发酵工艺优化及品质与风味评价

本研究以膨化藜麦粉、牛奶为原料,选取嗜热链球菌6063、保加利亚乳杆菌6064、动物双歧杆菌B15混合发酵藜麦牛奶,在单因素实验基础上采用正交试验对发酵工艺进行优化,并探究膨化藜麦粉的添加对酸奶品质及风味的影响。结果表明,三种益生菌发酵藜麦酸奶的最佳菌种配比为嗜热链球菌:保加利亚乳杆菌:动物双歧杆菌=2:1:2,最佳工艺为接菌量2%（v/v）,藜麦添加量5%（w/v）,发酵温度41 ℃。在此条件下,藜麦酸奶风味良好,乳酸菌总活菌数达到109 CFU/mL。从发酵开始至贮藏21 d结束,藜麦酸奶可滴定酸度始终高于纯酸奶,乳酸菌总活菌数低于纯酸奶。藜麦酸奶成品的蛋白质、多酚及黄酮含量分别比纯酸奶提高了88.67%、166.67%、284.62%。乳酸、甲酸、乙酸、丙酮酸含量分别为5.80、1.42、0.11、0.01 mg/g。利用气相色谱-质谱联用仪（GC-MS）进行风味分析,藜麦酸奶中共检出74种挥发性风味物质,与纯酸奶相比,新产生30种挥发性风味物质。研究结果将为藜麦酸奶新产品的开发提供工艺依据,进一步促进藜麦的应用与开发。     

麦胚乳酸菌发酵饮品的工艺优化及品质分析

以麦胚为原料,利用复合乳酸菌发酵研制麦胚乳酸菌发酵饮品。以离心沉淀率为评价指标,优化稳定剂含量;采用单因素试验及正交试验,优化麦胚乳酸菌发酵饮品工艺条件,并测定其理化指标。结果表明,最佳稳定剂为羟丙基二淀粉磷酸酯0.5%、琼脂0.2%;最佳发酵工艺条件为嗜热链球菌（Streptococcus thermophiles）、保加利亚乳杆菌（Lactobacillus bulgaricus）、乳双歧杆菌（Bifidobacterium lactis）、鼠李糖乳杆菌（Lactobacillus rhamnosus）、植物乳杆菌（Lactobacillus plantarum）、嗜酸乳杆菌（Lactobacillus acidophilus）、干酪乳杆菌（Lactobacillus casei）按照配比1∶1∶1∶1∶1∶1∶1制备发酵剂,接种量0.03%,发酵时间24 h,发酵温度 42 ℃。在此最佳条件下,麦胚乳酸菌发酵饮品为乳酪色,质地黏稠,口感细腻,总蛋白3.53 g/100 g,总膳食纤维2.12 g/100 g,维生素E 1.28 mg/100 g,亚油酸669 mg/100 g,α-亚麻酸75.8 mg/100 g,乳酸活菌数2.1×109 CFU/mL。     

Interactions among lactic acid starter and probiotic bacteria used for fermented dairy products

Interactions among lactic acid starter and probiotic bacteria were investigated to establish adequate combinations of strains to manufacture probiotic dairy products. For this aim, a total of 48 strains of Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactococcus lactis, Lactobacillus acidophilus, Lactobacillus casei, and Bifidobacterium spp. (eight of each) were used. The detection of bacterial interactions was carried out using the well-diffusion agar assay, and the interactions found were further characterized by growth kinetics. A variety of interactions was demonstrated. Lb. delbrueckii subsp. bulgaricus was found to be able to inhibit S. thermophilus strains. Among probiotic cultures, Lb. acidophilus was the sole species that was inhibited by the others (Lb. casei and Bifidobacterium). In general, probiotic bacteria proved to be more inhibitory towards lactic acid bacteria than vice versa since the latter did not exert any effect on the growth of the former, with some exceptions. The study of interactions by growth kinetics allowed the setting of four different kinds of behaviors between species of lactic acid starter and probiotic bacteria (stimulation, delay, complete inhibition of growth, and no effects among them). The possible interactions among the strains selected to manufacture a probiotic fermented dairy product should be taken into account when choosing the best combination/s to optimize their performance in the process and their survival in the products during cold storage.     

小米乳酸菌发酵饮料胀罐微生物的分离及鉴定

目的 对小米乳酸菌饮料胀罐微生物进行分离和菌株鉴定,并对所分离菌株的产气特性进行研究。方法 使用孟加拉红琼脂培养基、营养肉汤琼脂培养基和改良MRS琼脂培养基对小米乳酸菌饮料胀罐微生物进行分离和纯化,根据菌落形态观察、菌体形态观察、16S rRNA 序列分析并结合基质辅助激光解析/电离飞行时间质谱(ALDI-TOF MS)技术对分离菌株进行鉴定。最后将分离菌株接种到无菌小米乳酸菌饮料产品中,培养观察产气现象。结果 共分离出6株细菌,编号为B001、B002、B003、B003、B005、B006,菌种鉴定结果均为副干酪乳杆菌(Lactobacillus paracasei),6株细菌均具有产气现象。结论 副干酪乳杆菌是引起本研究产品胀罐的主要原因。     

益生菌发酵蓝莓花色苷的代谢规律

选用嗜热链球菌、两岐双歧杆菌、植物乳杆菌进行纯种发酵含有蓝莓花色苷提取液的培养液,考察了益生菌发酵过程中活菌数、体外抗氧化性、有机酸、单体花色苷和酚酸等组分的变化情况。结果表明,发酵48 h后,三种益生菌的活菌数均增至8.0 lg CFU/mL左右,两岐双歧杆菌的活菌数最高(P<0.05),发酵后不同菌株发酵样的ABTS+·清除能力显著提升(P<0.05),其中两岐双歧杆菌发酵样品抗氧化能力最强(P<0.05);但植物乳杆菌发酵样品中乳酸含量最高(P<0.05);发酵过程中单体花色苷含量呈下降趋势,两岐双歧杆菌发酵样品花色苷组分与其它两株菌发酵的样品差异较大;绿原酸、对香豆酸和咖啡酸的含量总体呈现下降趋势,而没食子酸、丁香酸和阿魏酸含量呈上升趋势,主成分分析图中三株益生菌发酵样的分布差异较大。以上结果为进一步解释益生菌发酵代谢花色苷的机理提供了参考。