Inactivation of calcium-dependent lactic acid bacteria phages by phosphates

The capacity of three phosphates to interrupt the lytic cycle of four specific autochthonal bacteriophages of lactic acid bacteria used as starters was assayed. The phosphates used (polyphosphates A and B and sodium tripolyphosphate-high solubility [TAS]) were selected on the basis of their capacity to sequester divalent cations, which are involved in the lytic cycle of certain bacteriophages. The assays were performed in culture media (deMan Rogosa Sharpe and Elliker broths) and reconstituted (10%, wt/vol) commercial skim milk to which phosphates had been added at concentrations of 0.1, 0.3, and 0.5% (wt/vol). Phosphate TAS was the most inhibitory one, since it was able to inhibit the lytic cycle of all bacteriophages studied, in both broths and milk. In broth, polyphosphates A and B inhibited the lytic cycle of only two bacteriophages at the maximal concentration used (0.5%), whereas in milk, they were not capable of maintaining the same inhibitory effect.     

Effect of high hydrostatic pressure treatment on the viability and acidification ability of lactic acid bacteria

The effect of high pressures of 100–450 MPa combined with temperatures of 20–40 °C on Lactobacillus delbrueckii subsp. bulgaricus ACA-DC0105, Streptococcus thermophilus ACA-DC0022 and Lactococcus lactis ACA-DC0049 cell viability and acid production ability was studied. The rates of decrease in pH and cell viability were estimated for all the process combinations studied. The viability and acidification ability of the cells depended on the process conditions. More intense process conditions resulted in a lower number of viable cells and simultaneous reduction of lactic acid production correlated to lower rates of decrease of pH. Lb. bulgaricus appeared to be the microorganism most resistant to pressure, while Lc. lactis the most sensitive. Similar behaviour was observed for the acidification ability of these microorganisms, with Lc. lactis being least able to decrease the pH value of cheese. The HP-treated strains could be used as adjunct starters for cheese production.     

乳酸菌的益生特性及应用研究进展

活性乳酸菌产品因其良好的益生功效越来越受到人们的青睐,市场需求量不断增大。然而,受乳酸菌厌氧和热敏感性的限制,以及加工中加热和氧气胁迫的影响,产品中活性乳酸菌数量大幅度下降。进入体内的活性乳酸菌在胃液高酸性和肠道高胆汁酸等复杂环境的胁迫下进一步降低,严重影响产品的益生功效。为此,人们在提高乳酸菌对热、氧气、胃肠环境耐受性方面进行了大量研究,并开发了系列新技术,但缺乏彼此间的综合对比与分析。本文围绕如何提高乳酸菌的加工存活率,以及胃肠耐受性和肠道递送问题,对静电纺丝、静电喷雾、乳滴技术、多酚纳米盔甲、热诱导预处理等新技术的作用效果进行了总结与对比,以期为相关研究和技术应用提供参考。     

Thermophilic lactic acid bacteria phages isolated from Argentinian dairy industries

Sixty-one natural phages (59 of Streptococcus thermophilus and 2 of Lactobacillus delbrueckii subsp. bulgaricus) were isolated from Argentinian dairy plants from November 1994 to July 2000. Specifically, 17 yogurt samples (18% of all samples) and 26 cheese samples (79%) contained phages lytic to S. thermophilus strains. The number of viral particles found in samples ranged from 10(2) to 10(9) PFU/ml. The phages belonged to Bradley's group B or the Siphoviridae family (morphotype B1). They showed high burst size values and remarkably short latent periods. The results of this study show that phages were found more frequently in cheesemaking processes than in yogurt-making processes. The commercial streptococcus strains appeared to propagate more phages, whereas the natural strains propagated fewer phage strains. These results suggest that the naturally occurring cultures are inherently more phage resistant.     

分离自藏灵菇的乳酸菌的益生特性

从藏灵菇中分离纯化5株乳酸菌,初步鉴定2株为嗜酸乳杆菌,3株为乳酸乳球菌。选取其中2株菌研究其益生特性。结果表明,从藏灵菇中分离出的乳酸菌具有良好的益生特性,2株乳酸菌在pH值为4～6可生长良好;耐热范围为30～60℃;胆盐耐受性为0.1%～0.5%;发酵液对金黄色葡萄球菌、沙门氏菌、大肠杆菌等肠道病原菌有抑制作用;对抗生素有不同程度的耐药性。     

泡菜中益生性乳酸菌的筛选和鉴定

以筛选具益生特性的泡菜发酵剂为目的,从15个泡菜样品中通过比较溶钙圈的大小分离出59株产酸能力较强的菌株。通过测定发酵液的酸度以及耐酸性和耐胆汁盐试验,再从中筛选出2株产酸能力和在胃肠道生存能力都强的菌株A18和B17。经形态特征及16S rDNA序列分析,确定A18和B17均属于植物乳杆菌。     

In vitro testing of commercial and potential probiotic lactic acid bacteria

Probiotics are defined as live microorganisms which when administered in adequate amounts confer a health benefit on the host. The objective of this study was to investigate the diversity of selected commercial and potential probiotic lactic acid bacteria using common in vitro screening assays such as transit tolerance in the upper human gastrointestinal tract, adhesion capacity to human intestinal cell lines and effect on epithelial barrier function. The selected bacteria include strains of Lactobacillus plantarum, Lactobacillus pentosus, Lactobacillus farciminis, Lactobacillus sakei, Lactobacillus gasseri, Lactobacillus rhamnosus, Lactobacillus reuteri and Pediococcus pentosaceus. Viable counts after simulated gastric transit tolerance showed that L. reuteri strains and P. pentosaceus tolerate gastric juice well, with no reduction of viability, whereas L. pentosus, L. farciminis and L. sakei strains lost viability over 180min. All strains tested tolerate the simulated small intestinal juice well. The bacterial adhesion capacity to human intestinal cells revealed major species and strain differences. Overall, L. plantarum MF1298 and three L. reuteri strains had a significant higher adhesion capacity compared to the other strains tested. All strains, both living and UV-inactivated, had little effect on the epithelial barrier function. However, living L. reuteri strains revealed a tendency to increase the transepithelial electrical resistance (TER) from 6 to 24h. This work demonstrates the diversity of 18 potential probiotic bacteria, with major species and strain specific effects in the in vitro screening assays applied. Overall, L. reuteri strains reveal some interesting characteristics compared to the other strains investigated.     

益生乳酸菌在乳制品中的应用研究进展

益生乳酸菌是一类能利用碳水化合物发酵产生大量乳酸,且对宿主有益的微生物。乳制品在人类膳食结构中占有十分重要的地位,随着消费者对乳品品质和健康要求的提升,具有各种健康功能的益生乳酸菌在乳品中的应用及相关加工技术和功能产品的研发日益受到关注。通过添加益生乳酸菌等活性因子获得功能性乳制品,是增强乳品健康功效的有效方法。本文综述了近年来有关益生乳酸菌在发酵乳、干酪、乳饮料、冰淇淋和奶粉等乳制品中的应用研究现状,重点介绍了益生乳酸菌发挥功能的主要代谢产物酶类和胞外多糖的应用研究,包括在不同乳制品中益生乳酸菌及其产物发挥的作用,常用的益生乳酸菌菌株种类,生产加工过程中存在的主要问题及解决方法,为益生乳酸菌在乳制品中的应用开发提供参考。     

潜在益生乳酸菌分离和鉴定研究进展

利用益生菌提高人类健康水平是当前营养健康领域最具前景的方向之一。乳酸菌是食品中常用的益生菌,迄今为止,联合国粮食及农业组织（FAO）和世界卫生组织（WHO）推荐仅从人的胃肠道分离的乳酸菌可在食品中使用。而越来越多的研究从食品来源中分离出具有益生功能的菌株以扩大益生菌的选择范围。随着分子生物学和检测分析等技术的发展,基质辅助激光解吸电离-飞行时间质谱（MALDI-TOF MS）、全基因组测序、实时荧光定量聚合酶链式反应（RT-FQ-PCR）等技术可以鉴定到亚种及菌株水平上的益生菌。鉴于分离鉴定技术是益生菌安全规范应用的先决条件,该文综述了近年来针对潜在益生乳酸菌有效分离和鉴定技术的研究进展,以期为我国益生乳酸菌自主开发利用提供借鉴。     

Effects of rare sugar D-allulose on acid production and probiotic activities of dairy lactic acid bacteria

It has recently been reported that the rare sugar d-allulose has beneficial effects, including the suppression of postprandial blood glucose elevation in humans, and can be substituted for sucrose as a low-calorie food ingredient. To examine the applications of d-allulose in the dairy industry, we investigated the effects of d-allulose on the acid production of 8 strains of yogurt starter (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) and 4 strains of lactococci, including potential probiotic candidates derived from dairy products. Acid production by 2 L. delbrueckii ssp. bulgaricus yogurt starter strains in milk was suppressed by d-allulose, but this phenomenon was also observed in some strains with another sugar (xylose), a sugar alcohol (sorbitol), or both. In contrast, among the dairy probiotic candidates, Lactococcus lactis H61, which has beneficial effects for human skin when drunk as part of fermented milk, was the only strain that showed suppression of acid production in the presence of d-allulose. Strain H61 did not metabolize d-allulose. We did not observe suppression of acid production by strain H61 with the addition of xylose or sorbitol, and xylose and sorbitol were not metabolized by strain H61. The acid production of strain H61 after culture in a constituted medium (tryptone–yeast extract–glucose broth) was also suppressed with the addition of d-allulose, but growth efficiency and sugar fermentation style were not altered. Probiotic activities—such as the angiotensin-converting enzyme inhibitory activity of H61-fermented milk and the superoxide dismutase activity of H61 cells grown in tryptone–yeast extract–glucose broth—were not affected by d-allulose. d-Allulose may suppress acid production in certain lactic acid bacteria without altering their probiotic activity. It may be useful for developing new probiotic dairy products from probiotic strains such as Lactococcus lactis H61.     

益生乳酸菌对氟喹诺酮类药物耐药机制的研究进展

益生乳酸菌对抗菌药物的耐药性机制是生物医药研究的重要领域。概述了益生乳酸菌对氟喹诺酮类药物的3种耐药性机制(靶位突变、外排泵作用、质粒介导)及其可能出现的耐药机制。显示出国内关于益生乳酸菌对氟喹诺酮类药物的耐药机制研究甚少的现状,提出对益生乳酸菌的耐药机制进行全面深入的研究的重要性。     

luxS基因介导乳酸菌益生特性研究进展

一些乳酸菌具有高肠道耐受性、高粘附肠上皮细胞和产具有抑菌活性的细菌素等益生特性。某些乳酸菌的细菌素合成量、耐受性及黏附特性可以被诱导物-2（autoinduction-2,AI-2）提高,AI-2是通过甲基循环合成的一种信号分子。luxS基因可以编码合成LuxS蛋白,而LuxS蛋白是AI-2合成的关键酶,因此展开对luxS在细菌素合成量和耐受性、黏附特性方面的作用研究具有重要意义。该文通过讨论luxS基因在乳酸菌益生特性的研究现状,提出该研究领域中存在的问题以及发展趋势,从而为提高乳酸菌的益生特性提供理论依据。     

广西生榨米粉中益生乳酸菌的筛选及鉴定

通过革兰氏染色、过氧化氢酶试验、溶血性试验、耐酸、耐胆盐试验及体外抗氧化能力测定从广西本地传统发酵食品生榨米粉中筛选优良益生乳酸菌,对其生长情况及产酸性能进行测定,并通过分子生物学技术对其进行鉴定。结果表明,共分离得到146株γ-溶血乳酸菌,其中22株乳酸菌对pH 2.0和0.3%胆盐均具有良好耐受性,且乳酸菌PM20、PM22、FD13及PM44的综合体外抗氧化能力较好。4株菌株的生长及产酸趋势一致,其中菌株PM44的生长速度和产酸能力最强。经鉴定菌株PM20、PM22、FD13均为发酵乳杆菌（Lactobacillus fermentum）,菌株PM44为乳酸片球菌（Pediococcus acidilactici）。     

Comparative survival and evaluation of functional probiotic properties of spray-dried lactic acid bacteria

The effect of spray drying on the viability and retention of key probiotic properties like acid and bile tolerance and cholesterol assimilation of three probiotic lactic acid bacteria (LAB) ( Lactobacillus plantarum CFR 2191, Lactobacillus salivarius CFR 2158 and Pediococcus acidilactici CFR 2193) has been studied. More than 97% survival was exhibited by the three LAB (1% cell suspension) spray dried with maltodextrin and nonfat skimmed (NFSM) as carriers. LAB cultures spray dried with maltodextrin showed significantly greater ( P ≤  0.05) retention of functional properties than those with NFSM. The results highlight a cost-effective way of producing large quantities of selected probiotic cultures with functional properties for neutraceutical application.     

产粘乳酸菌对酸奶品质的影响

将产粘乳杆菌与不产粘的保加利亚乳杆菌、不产粘嗜热链球菌组合生产酸奶，结果表明：产粘菌生产的酸奶口感不同于非产粘菌生产的酸奶，而且凝固状态好，乳清析出少，能达到最佳效果。使用产粘的乳酸菌可以大大降低酸奶的乳清析出，所产生的胞外多糖能起到稳定剂的作用，可以强化凝胶结构，从而使乳千物质对缩水作用的影响降低。     

酸和盐胁迫对乳酸菌活性的影响

该研究以13株食品常见乳酸菌为研究对象,考察酸和盐胁迫对其活性的影响,探究这13株乳酸菌的最适酸、盐生长条件及耐酸、耐盐能力。结果表明,适当的低盐浓度（1%、2%）对部分乳酸菌的生长有促进作用,植物乳杆菌和副干酪乳杆菌的最适盐浓度为1%,保加利亚乳杆菌和嗜酸乳杆菌的最适盐浓度为2%;盐浓度进一步增大,13株乳酸菌的生长受到抑制;盐浓度为10%时,除戊糖片球菌、乳酸乳球菌乳酸亚种、嗜热链球菌和乳酸片球菌外,其余9株菌的生长抑制率都＞95%。当pH值为5～7时,13株乳酸菌都生长良好,pH=6时,7株乳酸菌活性均最高;随着酸胁迫的增强,13株乳酸菌生长受抑制程度增大;pH为1～3时,13株乳酸菌的生长抑制率都＞90%,高盐及高酸对9株杆菌的抑制作用强于4株球菌。     

高抗氧化乳酸菌的筛选鉴定

为了寻找天然的抗氧化剂,对18株乳酸菌的菌体细胞和无细胞提取物的DPPH自由基清除能力、羟自由基的清除能力、超氧阴离子自由基的清除能力、还原能力、抗脂质过氧化能力等5项抗氧化指标进行比较,并对高抗氧化活性的乳酸菌进行抗性筛选,鉴定出符合要求的菌株。结果表明:18株乳酸菌具有不同的抗氧化能力,编号为L4、L5、L8、L14、L18的菌株具有较高的抗氧化活性,其中,菌株L14抗人工胃液和胆盐的能力较强,有作为天然抗氧化剂的应用前景。结合生化特性鉴定和16S r DNA鉴定,确定其为类干酪乳杆菌类干酪亚种(Lactobacillus paracasei subsp.paracasei)。     

Pressure-resistant acclimation of lactic acid bacteria from a natural fermentation product using high pressure

Twenty colonies of Pediococcus acidilactici, Lactobacillus brevis, Lactiplantibacillus plantarum and Latilactobacillus curvatus were isolated from fermentation broth of pickled pepper by using MRS agar. The strains were acclimated using high pressure from 300 to 400 MPa for 1 to 5 min. For one generation of acclimation, the survived cells after first high pressure treatment were then activated by culturing on MRS agar for two days for the next high pressure treatment. After acclimation for 30 generations, the survival ratio of L. plantarum and L. curvatus was increased by 138 and 1222 times at 400 MPa for 1 and 3 min, respectively. Weibull model could appropriately fit the survival curves of L. plantarum and L. curvatus treated by high pressure. The time to the first decimal reduction of these two strains after acclimation was 3.5 and 1.8 times, respectively, longer than before acclimation at 300 MPa. From electron microscopy photographs, the acclimated L. curvatus was more curved, smaller and its cell wall was thicker than the unacclimated L. curvatus. After HPP treatment, L. plantarum and L. curvatus have higher cell integrity, smoother cell surface, more uniform protoplasm and smaller cavities in the cell protoplasm compared with unacclimated strains, suggesting that high pressure acclimation introduced some modification to bacteria intrinsic structures and increased their pressure resistance. Moreover, the fermentation performance including glucose utilization capacity and lactic acid production of the acclimated L. curvatus was respectively improved by 9.6% and 9.4% at 37 °C for 24 h in MRS medium.     

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.     

一株高产酸乳酸菌的分离鉴定及生物学特性研究

利用MRS选择性培养基从豆清发酵液中分离筛选乳酸菌。经初筛得到5株乳酸菌,通过比较不同乳酸菌在同一条件下的p H和产乳酸量复筛得到一株产酸较高、p H较低的优势产酸乳酸菌,对其编号为M-6。根据菌株的形态学观察、生理生化试验以及16S r RNA基因序列分析鉴定,初步将该菌株鉴定为肠膜明串珠菌(Leuconostoc mesenteroides strain M-6)。对其进行生物学特性研究,发现菌株M-6的最适生长温度在30℃,适宜p H范围为5.5~8.5,菌株在该温度下发酵20 h后菌液OD600nm高达1.2917,p H低至4.16(总酸),且菌株M-6有一定的耐盐受性,当Na Cl浓度大于3%时,菌株生长受到明显抑制。因分离得到的肠膜明串珠菌M-6具有较强的产酸能力,可利用其发酵制备豆腐凝固剂,在豆制品生产领域具有良好的应用前景。