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。     

西藏灵菇中分离两株干酪乳杆菌及其益生特性比较研究

从西藏灵菇中筛选出28株革兰氏染色阳性菌,通过生理生化特征和16S rDNA序列同源性分析分离筛选出2株干酪乳杆菌（Lactobacillus casei）G1、G2,对其进行益生特性比较研究。结果表明,菌株G1、G2均具有较快的产酸能力;均可以在较大温度范围中生长,最适温度为40 ℃左右;具有较强的耐酸性;菌株G2耐胆盐能力明显高于菌株G1;菌株G1、G2对金黄色葡萄球菌（Staphylococcus aureus）、大肠杆菌（Escherichia coli）和沙门氏菌（Salmonella）均具有抑制作用,对大肠杆菌的抑制效果较为明显;菌株G1、G2对抗生素敏感性存在差异,对庆大霉素、诺氟沙星和氯霉素均产生耐药性,呈现多重耐药性。     

Pediococcus acidolactici and Pediococcus pentosaceus isolated from a rainbow trout ecosystem have probiotic and ABF1 adsorbing/degrading abilities in vitro

ABSTRACT

Probiotics are being used in biological control of bacterial pathogens, as an alternative to antibiotics, to improve health and production parameters in fish farming. Fish farming production is severely affected by aflatoxins (AFs), which are a significant problem in aquaculture systems. Aflatoxins exert substantial impact on production, causing disease with high mortality and a gradual decline of reared fish stock quality. Some aspects of aflatoxicosis in fish, particularly its effects on the gastrointestinal tract, have not been well documented. The aim of the present study was to evaluate probiotic properties of lactic acid bacterial (LAB) strains isolated from rainbow trout intestine and feed. Moreover, AFB₁-binding and/or degrading abilities were also evaluated to assess their use in the formulation of feed additives. Growth at pH 2, the ability to co-aggregate with bacterial pathogens, inhibition of bacterial pathogens, and determination of the inhibitory mechanism were tested. Aflatoxin B₁ (AFB₁) adsorption and degradation ability were also tested. All strains were able to maintain viable (10⁷ cells ml^–1) at pH 2. Pediococcus acidilactici RC001 and RC008 showed the strongest antimicrobial activity, inhibiting all the pathogens tested. The strains produced antimicrobial compounds of different nature, being affected by different treatments (catalase, NaOH and heating), which indicated that they could be H₂O_2, organic acids or proteins. All LAB strains tested showed the ability to coaggregate pathogenic bacteria, showing inhibition percentages above 40%. Pediococcus acidilactici RC003 was the one with the highest adsorption capacity and all LAB strains were able to degrade AFB₁ with percentages higher than 15%, showing significant differences with respect to the control. The ability of some of the LAB strains isolated in the present work to compete with pathogens, together with stability against bile and gastric pH, reduction of bioavailability and degradation of AFB₁, may indicate the potential of LAB for use in rainbow trout culture.     

Metabolic activities of Lactobacillus spp. strains isolated from kefir

A total of 21 strains of Lactobacillus species were isolated from Turkish kefir samples, in order to select the most suitable strains according to their metabolic activities including probiotic properties. As a result of the identification tests, 21 Lactobacillus isolates were identified as L. acidophilus (4%), L. helveticus (9%), L. brevis (9%), L. bulgaricus (14%), L. plantarum (14%), L. casei (19%) and L. lactis (28%). The amount of produced lactic acid, hydrogen peroxide, proteolytic activity, and acetaldehyde productions of Lactobacillus spp. were determined. Different amounts of lactic acid were produced by strains studies; however, lactic acid levels were 1.7-11.4 mg/mL. All strains produced hydrogen peroxide. L. bulgaricus Z14L strain showed no proteolytic activity, L. casei Z6L strain produced the maximum amount (0.16 mg/mL) of proteolytic activity. Acetaldehyde concentration produced in Lactobacillus strains ranged between 0.88-3.52 microg/mL.     

Cellular injuries of spray-dried Lactobacillus spp. isolated from kefir and their impact on probiotic properties

The injuries caused by spray drying (SD) of three potential probiotic lactobacilli isolated from kefir grains and the impact on some probiotic properties, were evaluated. Results demonstrated that Lactobacillus plantarum 83114 and L. kefir 8321 showed a slight reduction of viability (0.11 and 0.29 log CFU/ml respectively) after SD process, and L. kefir 8348 was found to be more sensitive to the process with a reduction in viability of 0.70 log CFU/ml. Neither membrane damage, evaluated by increased sensitivity to NaCl, lysozyme, bile salt and penicillin G, nor changes in acidifying activity in MRS and milk by lactobacilli were detected after SD. L. plantarum 83114 and L. kefir 8321 after SD did not lose their capacity to adhere to intestinal cells. Nevertheless, L. kefir 8348 showed a significant loss of adhesion capacity after SD. In addition, rehydrated spray-dried L. kefir 8321 retained the ability to protect against Salmonella invasion of intestinal cells. This effect was observed when L. kefir is co-incubated with Salmonella before invasion assay.This work shows that the membrane integrity evaluated by indirect methods and some probiotic properties of lactobacilli isolated from kefir did not change significantly after SD, and these powders could be used in functional foods applications.     

蒙古国传统发酵酸马奶(Koumiss)中乳杆菌潜在益生特性的研究

从分离自蒙古酸马奶样品(30株乳杆菌)中筛选出3株能够在pH值为3.0的酸性环境中生长良好的乳杆菌,分别为Lb.acidophilus MG2-1,Lb.acidophilus MG5-1和Lb.acidophilusMG1-12,但对pH值为2.0的人工胃液几乎不耐受。发酵乳中的乳杆菌对人工胃肠液的耐受性高于单纯菌体(P﹤0.05),乳蛋白对菌体起到一定的保护作用。3株Lb.acidophilus对胆汁都具有一定的耐受性,但所能耐受最高胆盐浓度有所差异;Lb.acidophilus MG2-1,Lb.acidophilus MG5-1和Lb.acidophilus MG1-12能耐受的最高胆盐浓度分别为6、4、3g/L;这3株Lb.acidophilus体外降胆固醇之间存在显著差异(P﹤0.05),其中Lb.acidophilus MG2-1对介质中胆固醇的脱除量达50%以上。     

In vitro probiotic evaluation of potential antioxidant lactic acid bacteria isolated from idli batter fermented with Piper betle leaves

Lactic acid bacteria isolated from food sources can be introduced as probiotics because of their health‐promoting and nonpathogenic characteristics. Eight lactic acid bacteria from idli batter fermented with Piper betle leaves were selected to screen the antioxidant potential and biofilm‐forming ability. Five isolates exhibited good antioxidant potential and biofilm formation and showed antagonistic activity against the pathogenic biofilm‐forming Staphylococcus aureus. Further, in vitro probiotic properties of five isolates exhibited excellent acid tolerance, bile tolerance, simulated gastrointestinal juice tolerance, auto‐aggregation and co‐aggregation and hydrophobicity and showed resistance to antibiotics such as ciprofloxacin, nalidixic acid, norfloxacin and colistin. Isolates also showed positive bile salt hydrolase activity, cholesterol assimilation, β‐galactosidase production and mucin binding ability. The isolate KJBB10 exhibited good adhesion and pathogen exclusion percentage with human colon cancer cells (HCT‐15 and HT‐29). Hence, the isolates KJBB10, KJBB56 and KJBC06 can be used as beneficial probiotic starter cultures for the formulation of functional foods.     

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。     

嗜酸乳杆菌和双歧杆菌益生特性的研究

以两株嗜酸乳杆菌和三株动物双歧杆菌为研究对象,详细研究它们的耐胃酸性、耐胆汁盐性、抑菌性和抗药性。结果表明:五株益生菌具有良好的耐酸耐胆汁盐性,双歧杆菌的耐酸性较强于嗜酸乳杆菌;抑菌性和抗药性结果表明:五株益生菌对致病菌大肠杆菌、金黄色葡萄球菌和沙门氏菌均有抑制作用,嗜酸乳杆菌的抑菌性较强于双歧杆菌;五株益生菌对三种抗生素乙酰螺旋霉素、链霉素和青霉素均敏感,敏感程度各异。     

Phenotypic and genetic diversity of Lactococcus lactis and Enterococcus spp. strains isolated from Northern Spain starter-free farmhouse cheeses

To evaluate a previous phenotypic classification of lactococci, 39 presumed lactococcal strains were classified by molecular techniques. The strains were also subjected to several typing techniques to estimate the phenotypic and genetic diversity present in original populations from starter-free farmhouse cheeses. Partial Amplified rDNA Restriction Analysis (partial ARDRA) with either restriction enzyme MboII or HhaI divided these isolates into four distinctive groups. Sequencing of representative amplicons identified 29 isolates as belonging to Lactococcus lactis subsp. lactis (24) and Lactococcus lactis subsp. cremoris (5). The remaining 10 isolates were shown to be Enterococcus durans (8) and Enterococcus faecalis (2), which were misclassified by the traditional tests. Thus, partial ARDRA was successfully used to classify wild Lactococcus-like strains into Lactococcus and Enterococcus species. The technique also allowed differentiation of L. lactis strains at subspecies level. The 29 strains of L. lactis showed five different fermentation profiles, four distinct Random Amplification of Polymorphic DNA (RAPD) profiles, and 14 unrelated profiles by both Restriction Fragment Length Polymorphism analyzed by Pulsed Field Gel Electrophoresis (RFLP-PFGE) and Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Using the same techniques, the 10 enterococcal strains showed four fermentation profiles, four RADP, and six by RFLP-PFGE and SDS-PAGE, respectively. Several typing techniques, especially RFLP-PFGE and SDS-PAGE, revealed wide phenotypic and genetic variability in both the lactococcal and enterococcal isolates. Two simple, rapid and cheap techniques (partial ARDRA and SDS-PAGE) are proposed as reliable tools for the classification and typing of new lactococcal-like isolates.     

Health-promoting benefits of low-fat akawi cheese made by exopolysaccharide-producing probiotic Lactobacillus plantarum isolated from camel milk

Lactic acid bacteria isolated from camel milk exhibit remarkable probiotic and exopolysaccharide (EPS)-producing characteristics. The health-promoting benefits of exopolysaccharide-producing probiotic Lactobacillus plantarum isolated from camel milk used for making low-fat akawi cheese were investigated. Three low-fat akawi cheeses were made using traditional culture (non-EPS-producing, EPS^?), commercial EPS-producing (MEPS⁺), and camel milk EPS-producing (CEPS⁺) cultures. α-Amylase and α-glucosidase inhibitory activities, antioxidant activities, angiotensin-converting enzyme (ACE) inhibition, and antiproliferative activity were determined. Cheese made with CEPS⁺ culture exhibited comparable α-amylase inhibition to that of cheeses made with MEPS⁺. Scavenging rates of cheese made with EPS⁺ cultures were higher than those of cheese made with EPS^? cultures. The percentage of α-glucosidase inhibition ranged from >45% at 0 d to ～55% at 21 d of storage. After 7 d of storage, the scavenging rate in CEPS⁺ cheese increased >60% by ABTS assay [2,2'-azino-bis(3-ethylbenzo-thiazoline-6-sulfonic acid)] and >20% by DPPH assay (1,1-diphenyl-2-picrylhydrazyl). Throughout storage, cheese made with EPS⁺ cultures showed higher ACE-inhibition activity compared with EPS^? cultures. Cheese made with CEPS⁺ showed ACE inhibition >70% after 7 d of storage. Antiproliferation activity of CEPS⁺ cheese increased from 38 to 48% during 7 d of storage and was maintained above 45% with prolonged storage. Low-fat akawi cheese produced with these cultures exhibited similar or greater health-promoting benefits compared with cheese made using commercial starter cultures. Therefore, incorporation of these cultures in food is promising for commercial uses.     

Evaluation of exopolysaccharide production by Leuconostoc mesenteroides strains isolated from wine

ABSTRACT:  Exopolysaccharide (EPS)-producing lactic acid bacteria are responsible for the alteration of wine and other fermented beverages. The potential to produce EPS was investigated for Leuconostoc mesenteroides strains isolated from Spanish grape must and wine. Most strains were able to produce EPS from sucrose containing media. Based on their EPS-producing phenotype and on their EPS monosaccharide composition, the L. mesenteroides strains analyzed could be arranged in 2 groups. One group comprises mucoid strains producing a glucan polymer, and the other group includes strains producing a fructan polymer. The presence of a glucosyltransferase encoding gene in the glucan producing L. mesenteroides strains was assayed by PCR. Two primer sets, PF1-PF8 and GTFF-GTFR, were used to amplify internal fragment of known glucosyltransferase genes. None of the glucan-producing strains gave a positive amplicon by the primer sets used. Therefore, new tools need to be developed to broaden the range of potentially spoiling agents detected by PCR in fermented beverages.     

一株强抑菌活性植物乳杆菌的分离及益生性能研究

以健康黔北黑猪肠道黏膜内容物为原料,采用厌氧罐培养法分离出肠道细菌,结合抑菌试验筛选出其中具有强抑菌活性菌种进行分子生物学鉴定,并对其耐受性及抑菌物质的稳定性进行研究。结果表明,该菌株被鉴定为植物乳杆菌（Lactobacillus plantarum）,对金黄色葡萄球菌ATCC6538、大肠埃希氏菌ATCC25922和鼠伤寒沙门氏菌ATCC14028等指示菌具有强抑制作用,其发酵上清液对3种指示菌的最大抑菌圈直径分别约为44.7 mm、23.1 mm和31.8 mm;菌株发酵上清液抑菌物质耐高温,120 ℃处理30 min抑菌活性无显著变化（P＞0.05）;胰蛋白酶处理后,上清液的抑菌圈直径显著减小（P＜0.05）,而过氧化氢酶和胃蛋白酶处理后抑菌圈直径无显著变化（P＞0.05）,表明主要抑菌物质为有机酸和活性肽;菌株经pH 2、0.3%猪胆盐或人工胃肠液处理6 h后存活率分别为0.06%、0.61%、40.80%和25.65%,显示出良好的益生性能。     

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）。     

植物乳杆菌C21胞外多糖的分离纯化及其结构修饰

为研究乳酸菌胞外多糖的结构和性质,采用乙醇沉淀、DEAE- 纤维素离子交换柱层析和 Sepharose CL-6B凝胶过滤层析方法从植物乳杆菌(Lactobacillus plantarum) C21 SDM 液体培养基中分离纯化得到一种均一组分的胞外多糖,命名为PCPc。采用离子色谱和红外光谱法对其单糖组成和结构进行初步分析,结果表明,PCPc 主要由半乳糖和葡萄糖两种单糖组成,其物质的量比为1:2。并对PCPc 多糖进行化学修饰,获得了其硫酸化和磷酸化衍生物。     

Stimulation of Cadaverine Production by Foodborne Pathogens in the Presence of Lactobacillus,Lactococcus, and Streptococcus spp.

Abstract: The effect of Lactobacillus plantarum (FI8595), Lactococcus lactis subsp. cremoris MG 1363), Lactococcus lactis subsp. lactis (IL 1403), and Streptococcus thermophilus on cadaverine and other biogenic amine production by foodborne pathogens was investigated lysine decarboxylase broth. Both of lactic acid bacteria and foodborne pathogens used (especially Staphylococcus aureus, E. coli, Lc. lactis subsp. lactis and Lb. plantarum) had an ability to convert aminoacids into biogenic amine. The conversion of lysine into cadaverine was the highest (167.11 mg/L) by Lactobacillus spp. Gram‐positive bacteria generally had a greater ability to produce cadaverine with corresponding value of 46.26, 53.76, and 154.54 mg/L for Enterococcus faecalis, S. aureus, and Listeria monocytogenes, respectively. Significant variations on biogenic amine production were observed in the presence of lactic acid bacteria strains (P < 0.05). The role of lactic acid bacteria on biogenic amine production by foodborne pathogens varied depending on strains and specific amine. Cadaverine accumulation by Enterobactericeae was increased in the presence of lactic acid bacteria strains except for St. thermophilus, which induced 2‐fold lower cadaverine production by S. Paratyphi A. Lc. lactis subsp. lactis and Lc. lactis subsp. cremoris induced 10‐fold higher increases in histamine for E. coli and K. pneumoniae, respectively. Lactic acid bacteria resulted in strong increases in cadaverine production by P. aeruginosa, although remarkable decreases were observed for histamine, spermidine, dopamine, agmatine, and TMA in the presence of lactic acid bacteria in lysine decarboxylase broth . The result of the study showed that amine positive lactic acid bacteria strains in fermented food led to significant amine accumulation by contaminant bacteria and their accumulation in food product may be controlled by the use of proper starters with amine‐negative activity. Practical Application: Foodborne pathogens and certain lactic acid bacteria are particularly active in the production of biogenic amines. Most of the strains of bacteria possess more than 1 amino acid decarboxylase activity under lysine enrichment culture conditions. Lactic acid bacteria strains had a significant role on increase putrescine accumulation by foodborne pathogens. The increased production of biogenic amines in mixed culture is the result of presence of amine positive lactic acid bacteria strains. The addition of a proper selected starter culture with amine‐negative activity is advisable to produce safer fermented food with low contents of biogenic amines.     

Synbiotic potential of carrot juice supplemented with Lactobacillus spp. and inulin or fructooligosaccharides

BACKGROUND: The influence of the addition of two probiotic strains, Lactobacillus rhamnosus and Lactobacillus bulgaricus, and the prebiotic components inulin and fructooligosaccharides on carrot juice was investigated in order to evaluate the possibility of producing a functional vegetal beverage. RESULTS: Both bacterial strains were capable of growing in carrot juice, reaching nearly 5 × 10⁹ colony‐forming units after a 48 h fermentation, and the pH was reduced to 3.5–3.7 or below. The viable cell counts of the two lactobacilli in the fermented juice after 4 weeks of storage at 4 °C, demonstrated good survival of the two strains at low pH. Some biochemical characteristics of the fermented juice, such as β‐carotene content and antioxidant activity, were also preserved, indicating that the metabolism of the Lactobacillus spp. did not degrade these nutritional components after 4 weeks of storage at 4 °C. The positive effect was more evident when the juice was inoculated with Lactobacillus rhamnosus. The presence of inulin and fructooligosaccharides did not alter the cell counts or the biochemical characteristics of the fermented juice. CONCLUSION: We propose this as a health juice and functional beverage for vegetarians or consumers who are allergic to dairy products. Copyright © 2008 Society of Chemical Industry     

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.     

Functional properties of Lactobacillus strains isolated from kimchi

The objective of this study was to evaluate the functional properties of lactic acid bacteria (LAB) from kimchi, a traditional Korean fermented vegetable product generally consumed raw as a side-dish with practically every meal.Twelve mild acid producing facultatively heterofermentative Lactobacillus strains were selected for their potential as starter cultures for fermentation of kimchi, and evaluated for their functional properties. Eleven strains were identified as Lactobacillus sakei and one as Lactobacillus plantarum. The strains identified as L. sakei differed in some physiological features; of particular interest was the fact that 9 of these strains produced L(+) lactic acid from glucose in presence of acetate.All strains were able to survive gastrointestinal conditions simulating stomach and duodenum passage. In addition, they showed higher adherence to HT-29 cells than Lactobacillus rhamnosus GG, a commercial probiotic strain used worldwide. These strains also showed antimicrobial activity against a number of food-borne pathogens. Their ability to lower cholesterol was demonstrated by BSH (bile salt hydrolytic) activity, and cholesterol assimilation tests in vitro. The results suggest the probiotic potential of these strains for use in kimchi fermentation.