3株益生菌体外降胆固醇能力及体内降血脂效果评价

为探讨动物双歧杆菌LPL-RH、长双歧杆菌TTF及植物乳杆菌LPL-1作为降血脂益生菌的潜力,测定3株益生菌的体外胆固醇脱除率和胆盐水解酶活力,同时以益生菌灌胃高脂饮食大鼠6周后测定大鼠血脂含量变化。研究结果表明:菌株LPL-RH、TTF、LPL-1的体外胆固醇脱除率分别为23.8%、24.5%、20.9%,无显著差异(P0.05);且3株菌均有胆盐水解酶活性。菌株LPL-RH、TTF、LPL-1干预分别使高脂大鼠血清胆固醇含量分别降低17.3%、11.4%、10.1%;血清甘油三酯含量降低19.1%、16.8%、11.5%;血清低密度脂蛋白胆固醇含量降低24.0%、20.0%、10.0%;血清高密度脂蛋白胆固醇含量升高10.3%、5.2%和1.7%。综合而言,3株益生菌在体内对高脂饮食大鼠血脂有调节作用,均有开发为降血脂益生菌的潜力,其中动物双歧杆菌LPL-RH的体内降脂能力最为突出。     

植物乳杆菌的降胆固研究

对植物乳杆菌MA2的体外和体内降胆固醇活性进行了研究,结果显示:培养基中添加的胆固醇的初始浓度对植物乳杆菌MA2的胆固醇去除率影响显著,当胆固醇的终质量浓度为30mg/dL,不添加胆盐的条件下,菌株MA2的胆固醇去除率达到(55±4.5)%。死菌体和休眠菌体也可去除培养基中的胆固醇,且菌体细胞浓度的影响显著。添加胆盐对菌株MA2的胆固醇去除率影响显著。体外实验表明植物乳杆菌MA2通过吸附胆固醇,进而将胆固醇吸入菌体细胞中来降低培养基中的胆固醇。MA2菌株可显著降低大鼠血清总胆固醇、甘油三酯的水平,表明其作为降血脂的益生菌的应用潜力。     

添加植物乳杆菌的低脂干酪降胆固醇作用

为研究益生菌在干酪中应用的降胆固醇活性,以菌株胆固醇降解率、胆盐水解酶活力及模拟胃肠道耐受性为评价指标,筛选出具有降胆固醇活性的益生菌,并添加到低脂干酪(脂肪含量为1.0%)中,灌胃小鼠35 d,测定小鼠血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)含量、动脉硬化指数(AI)及肝脏、粪便中TC、TG含量。结果表明,植物乳杆菌KLDS 1.0320降胆固醇特性优于其他菌株,胆固醇降解率达到63.72%,模拟胃肠道消化后存活率为82.21%,胆盐水解酶活力为0.71 U/mL。与高脂模型组相比,灌胃添加植物乳杆菌KLDS 1.0320干酪小鼠的TC、TG、LDL-C及AI分别下降21.23%、19.30%、31.73%和35.05%,肝脏中TC、TG显著降低,粪便中TC、TG显著升高(p<0.05)。实验获得了1株具有降胆固醇能力的植物乳杆菌KLDS 1.0320,以干酪为载体在动物体内仍具有降低血清胆固醇的作用,说明其具有应用于干酪发挥降胆固醇活性的潜力。     

四株益生菌对高脂血症仓鼠降血脂作用的研究

目的:观察不同益生菌菌株对实验性高脂血症仓鼠脂代谢的影响。方法:利用高脂饲料诱导仓鼠高脂血症模型,给仓鼠灌胃益生菌菌液,测定血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)的含量及肝脏组织中TC、TG的含量。结果:四株益生菌均可有效降低高脂膳食仓鼠血清TC、TG、非高密度脂蛋白胆固醇(non-HDL-C)含量及动脉粥样硬化指数(AI)值,提高HDL-C含量,还可以显著降低肝脏中TC、TG含量。结论:四株益生菌均能有效改善仓鼠血脂代谢,预防和减少动脉粥样硬化的发生。鼠李糖乳杆菌降低高脂饮食仓鼠血清TC、TG含量的作用最强,嗜酸乳杆菌、鼠李糖乳杆菌升高血清HDL-C含量的作用最强,但嗜酸乳杆菌降低肝脏中TC含量的作用最强。     

六堡茶乳酸菌多样性及其降胆固醇特性分析

探究六堡茶渥堆发酵过程中乳酸菌多样性并筛选具有降胆固醇活性菌株，旨在评价其作为潜在益生菌的体外降胆固醇能力，预防人体因饮食不均衡导致的高血脂症。采用高通量测序、纯培养法分析不同渥堆发酵时期六堡茶中乳酸菌多样性，并对其体外降胆固醇能力、胃肠耐受性进行研究。结果表明，从25 个样品中共检测到乳酸菌4 个属（乳杆菌属（Lactobacillus）、魏斯氏菌属（Weissella）、乳球菌属（Lactococcus）和肠球菌属（Enterococcus））5 个种，其中乳杆菌属为优势菌。共分离得到76 株乳酸菌，包括55 株戊糖乳杆菌（Lactobacillus pentosus）、3 株铅黄肠球菌（Enterococcus casseliflavus）、5 株类肠膜魏斯氏菌（Weissella paramesenteroides）、8 株希腊魏斯氏菌（Weissella hellenica）、5 株台湾乳球菌（Lactococcus taiwanensis）。通过乳酸菌体外降胆固醇实验，发现胆固醇去除率为0%～30%，其中戊糖乳杆菌L131的去除胆固醇率最高，为（29.56±0.37）%；在pH值分别约为3.0、8.0的人工胃液、人工胰液中，L131有最高存活率，分别为（92.70±0.71）%和（77.54±4.81）%，显著高于其他菌株（P＜0.05）。本研究探明六堡茶渥堆发酵过程中乳酸菌的多样性，且发现具有降胆固醇特性的优良乳酸菌株，为健康黑茶产品和茶源益生菌的开发提供理论基础。     

海带源降亚硝酸盐和胆固醇益生乳酸菌的筛选与鉴定

海带是天然的保健食品,本研究以淡干海带为原料,从中筛选出具有较强的降亚硝酸盐和胆固醇能力的优良菌株。采用盐酸萘乙二胺法测定菌株在MRS液体培养基中亚硝酸盐的含量,邻苯二甲醛法测定菌株在MRS液体培养基中胆固醇的含量,再对筛出的菌株进行鉴定。结果表明:从淡干海带中分离出的27株菌株中,有7株菌株具有很强的降解亚硝酸盐能力,降解率均达到97%以上。进一步对这7株菌株降解胆固醇的能力进行筛选,结果表明其中3个菌株降解胆固醇能力较强分别为:55.02%(La-3)、61.21%(La-10)、56.85%(La-15);经过形态特征观察、生理生化实验和16S r DNA基因序列相似性分析,结果显示这7株菌株均为植物乳杆菌。本文结果还表明了相同的植物乳杆菌其降解胆固醇的能力存在差异,显示了物种的多样性。     

降血脂鼠李糖乳杆菌的筛选及其对高脂模型地鼠血脂的影响

该实验旨在筛选出一株具有较强降血脂功能的乳杆菌。试验采用的菌株分离自健康婴儿肠道,通过邻苯二甲醛比色法测定菌株降胆固醇能力、单试剂法测定降甘油三酯能力,并测定了其对酸及胆盐的耐受性,从中筛选出具有降胆固醇和降甘油三酯功能较强的菌株进行16S rDNA鉴定。通过建立高血脂地鼠模型,研究菌株体内降血脂能力。结果表明,鼠李糖乳杆菌HCS01-013的降胆固醇和甘油三酯能力较强,其胆固醇降解率达到35.40%;甘油三酯降解率达到44.32%;在0.3%和1.5%的胆盐环境下培养17 h的存活率分别为99.67%和96.79%,在pH 3.0和pH 2.0环境下培养17 h后存活率能达到88.76%和71.34%;黄金地鼠喂养高脂饲料建立的高脂地鼠模型后,喂养鼠李糖乳杆菌HCS01-013各剂量组与喂养生理盐水的模型对照组相比甘油三酯含量均明显降低。鼠李糖乳杆菌HCS01-013具有一定的降血脂功能,且耐逆环境能力较强,为该菌株相关研究及产品研发提供理论依据。     

益生菌发酵豆乳对高脂血症小鼠降血脂及肝损伤缓解的作用

研究干酪乳杆菌、植物乳杆菌和双歧杆菌在体外降胆固醇能力及三种益生菌混合发酵豆乳对高脂饮食小鼠血脂及肝损伤的影响。采用邻苯二甲醛法评价益生菌体外降胆固醇能力。采用C57BL/6N小鼠,并将其随机分为7组,即对照组;高脂组;干酪乳杆菌发酵豆乳干预组;植物乳杆菌发酵豆乳干预组;双歧杆菌发酵豆乳干预组;三种益生菌混合发酵豆乳干预组;未发酵豆乳干预组。连续喂养7周后,测定血清和肝脏中相关血脂指标,并观察肝脏组织病理学变化。结果表明:干酪乳杆菌、植物乳杆菌和双歧杆菌在体外降胆固醇能力分别为43.14%、46.27%和40.53%。益生菌混合发酵豆乳显著降低小鼠腹部和肾周脂肪指数,血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL),升高高密度脂蛋白(HDL)水平,降低肝脏TC、TG、丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平,并明显改善肝脏脂肪堆积。     

高效降胆固醇乳酸菌的筛选

研究了25株嗜酸乳杆菌在MRS-CHOL培养基中对胆固醇的降解量,降胆固醇的能力,以及对酸、胆盐的耐受性。实验表明:所有实验菌株均有降解培养物中胆固醇的能力,其中有6株实验菌对培养物中胆固醇的降解率大于40%,这6株菌对胆固醇的脱除效力也较高,同时对这6株菌的耐酸、耐胆盐,以及对胆固醇的脱除能力进行了比较,筛选出降胆固醇能力较强同时对酸和胆盐耐受性较强的嗜酸乳杆菌1株。     

猪源降胆固醇乳酸菌的分离鉴定

乳酸菌是一种重要的益生菌,具有降解胆固醇能力。作者从猪消化道中分离出31株疑似菌株,在高胆固醇培养基中培养后有18株菌株生长。利用磷硫铁法对体外降胆固醇进行了研究,结果表明,胆固醇降解率在0～47.26%,其中4株乳酸菌的胆固醇降解率大于30%,分别是r16、r53、r762和m661。通过细胞形态、生理生化试验和16SrDNA分子生物学鉴定,最终鉴定r16、r53和r762为植物乳杆菌,m661为鹑鸡肠球菌。     

Attachment of Salmonella spp. and Listeria monocytogenes to stainless steel,rubber and polytetrafluorethylene: the influence of free energy and the effect of commercial sanitizers

Bacteria and material surfaces were characterized with respect to their hydrophobicity and surface free energy using the contact angle method. Salmonella strains showed higher hydrophobicity and lower surface free energies than Listeria monocytogenes strains. Polytetrafluorethylene was the most hydrophobic material (and had the lowest surface free energy), followed by rubber and stainless steel. Bacteria attached in higher numbers to the more hydrophobic materials. Bacterial adherence could not be correlated with surface free energies or contact angles of bacteria, although L. monocytogenes strains attached in higher numbers than Salmonella strains to all of the materials tested. The cleaning of materials with commercial sanitizers resulted in a decrease of their contact angles (and an increase of their surface free energies), accompanied by a reduction in the number of adhered bacteria in comparison with the standard conditions. The degree of reduction in bacterial adherence varied with the bacteria, the substrate material and the sanitizer tested. Quaternary ammonium compounds were more effective against Salmonella attachment than L. monocytogenes attachment. Diethylenetriamine showed similar efficacy against attachment of both bacteria. Polytetrafluorethylene showed the greatest reduction in attachment after being washed with commercial sanitizers. It is concluded that stainless steel is less adherent than rubber or polytetrafluorethylene and should be preferred in the food industry when possible. On the other hand, polytetrafluorethylene seems to be more easily sanitized. Since effectiveness of sanitizers in the reduction of bacterial adherence was dependent upon the bacteria and the materials studied, the use of mixtures of sanitizers would help to control bacterial adherence in the food industry.     

果蔬发酵液中3株乳杆菌的体外功能特性研究

以果蔬自然发酵液中分离得到的3株乳杆菌（Lactobacillus）为对象,通过测定表面疏水性、自身凝聚率分析乳杆菌的表面特性;以DPPH自由基、ABTS自由基清除能力和铁还原能力为指标,分析3株乳杆菌的体外抗氧化能力差异;以降解胆固醇及亚硝酸盐为指标,评价乳杆菌的功能活性。结果表明:3株菌中,L3的表面疏水性和自身凝聚力均表现良好,分别为57.7%和95.2%（24 h）;3株菌的上清液具有很强的DPPH自由基清除率（>90%）、羟自由基清除率（>68%）及还原能力（>1.0）,其中L1和L3的上清液具有更强羟自由基清除率,近90%,L2则具有较强的抗脂质过氧化能力（>37%）;乳杆菌在48 h内几乎完全降解200 mg/L NaNO₂;3株菌上清液均有较好的胆固醇降解能力（>19%）,其活性物质主要为蛋白质和多糖类物质。综上所述,果蔬发酵液中分离所得的3株乳杆菌表现出了良好的益生特性,可为开发功能性发酵食品奠定坚实的基础。     

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.     

Selection of tropical lactic acid bacteria for enhancing the quality of maize silage

The objective of this study was to select lactic acid bacteria (LAB) strains isolated from silage and assess their effect on the quality of maize silage. The LAB strains were inoculated into aqueous extract obtained from maize to evaluate their production of metabolites and pH reduction. The ability to inhibit the pathogenic and silage-spoilage microorganisms’ growth was evaluated. Nine LAB strains that showed the best results were assessed in polyvinyl chloride experimental silos. The inoculation of the LAB strains influenced the concentration of lactic and acetic acids and the diversity of Listeria. The inoculation of silages with Lactobacillus buchneri (UFLA SLM11 and UFLA SLM103 strains) resulted in silages with greater LAB populations and improvements after aerobic exposure. The UFLA SLM11 and SLM103 strains identified as L. buchneri showed to be promising in the treatment of maize silage.     

Control of Listeria monocytogenes with combined antimicrobials on beef franks stored at 4 degrees C

Contamination of ready-to-eat meat products such as beef franks with Listeria monocytogenes has become a major concern for the meat processing industry and an important food safety issue. The objective of this study was to determine the effectiveness of combinations of antimicrobials as aqueous dipping solutions to control L. monocytogenes on vacuum-packaged beef franks stored at 4 degrees C for 3 weeks. Commercial beef franks were dipped for 5 min in three antimicrobial solutions: pediocin (6,000 AU), 3% sodium diacetate and 6% sodium lactate combined, and a combination of the three antimicrobials. Samples were then inoculated with 10(7) CFU/g of either four L. monocytogenes strains individually or a cocktail of the four strains, vacuum packaged, and stored at 4 degrees C for 3 weeks. Sampling was carried out at day 0 and after 2 and 3 weeks of storage. Individual strains, as well as the cocktail, exhibited different responses to the antimicrobial treatments. After 2 and 3 weeks of storage at 4 degrees C, pediocin-treated beef franks showed a less than 1-log reduction for all bacterial strains. Samples treated with the sodium diacetate-sodium lactate combination showed about a 1-log reduction after 2 weeks of storage for all strains and between a 1- and 2-log reduction after 3 weeks of storage, depending on the bacterial strain. When the three antimicrobials were combined, reductions ranged between 1 and 1.5 log units and 1.5 to 2.5 log units after 2 and 3 weeks of storage, respectively, at 4 degrees C. These results indicate that the use of combined antimicrobial solutions for dipping treatments is more effective at inhibiting L. monocytogenes than treatments using antimicrobials such as pediocin separately.     

Rhamnolipid and surfactin inhibit Listeria monocytogenes adhesion

The aim of this study was to assess the potential use of biosurfactants in inhibiting the Listeria monocytogenes strains adhesion to polystyrene surfaces. Surfactin and rhamnolipids were used. The adhesion profiles of 15 strains showed that most of these bacteria can be classified as moderate to strongly adherent. Both biosurfactants were able to reduce bacterial adhesion, and the effect was more pronounced against the strongly adherent strains. The most promising result was obtained for ATCC7644 strain, which showed an adhesion reduction of 84% for surfactin (30-h growth period). Rhamnolipids decreased the ATCC15313 adhesion by 82%. ATCC19112 adhesion was reduced by 53% for surfactin and purified rhamnolipid. Sodium dodecyl-sulphate was less effective than the biosurfactants, showing maximal adhesion reduction of 23% for 19112 and 7644. The purified rhamnolipid inhibited 100% of the growth of strongly adherent, suggesting that this surfactant can be exploited as a potential agent to control L. monocytogenes.     

复合乳酸菌发酵降低蛋黄液中胆固醇的工艺优化

为降低蛋黄液中胆固醇含量,选择出降胆固醇效果较好的3 种菌作为复合发酵菌种,将复合菌株接种到蛋黄液中,于恒温培养箱中培养,用直接皂化-比色法探讨了复合乳酸菌对蛋黄液中胆固醇降低率的影响,利用正交试验优化最佳发酵工艺参数。结果表明,最佳工艺条件为发酵温度40 ℃、发酵时间24 h、接种量3%、牛胆盐质量浓度3 g/L、混合菌种（嗜热链球菌、嗜酸乳杆菌、双歧杆菌）菌液体积比1∶2∶1,在此条件下,可使蛋黄中胆固醇降低率达31.5%。     

D-苏氨酸在低产硫化氢啤酒酵母选育中的应用

研究了酵母菌、麦芽汁中L-苏氨酸对发酵液后期硫化氢含量的影响。结果表明,不同的酵母产硫化氢的能力差异很大,使用菌种H的发酵液后期的硫化氢含量为84.24μg/L,使用菌种E的发酵液后期的硫化氢含量为3.45μg/L;麦芽汁中的L-苏氨酸可以促进发酵液中硫化氢的积累。通过筛选D-苏氨酸抗性突变株,可以提高获得低产硫化酵母的概率,初筛正突变率高达50%。在中试试验结果表明,与出发菌种相比较,突变菌种的发酵液后期的硫化氢含量降低34%。     

Identification of tet(M) in two Lactococcus lactis strains isolated from a Spanish traditional starter-free cheese made of raw milk and conjugative transfer of tetracycline resistance to lactococci and enterococci

Specific PCR and sequencing showed that a tet(M) gene was present in two tetracycline-resistant Lactococcus lactis strains isolated from a raw milk, starter-free cheese. Hybridisation experiments using as a probe an internal segment of the gene obtained by PCR associated tet(M) with plasmids of around the same size (30 kbp) in both strains. Molecular analysis of the tetracycline resistance loci, including the upstream and downstream regions of the genes, showed them to be identical to one other and to the tet(M) encoded by the conjugative transposon Tn916. Amplification of Tn916-derived segments suggested the transposon was complete in the two L. lactis strains. Further, curing of the tetracycline resistance was accompanied by a reduction in size of the plasmids comparable to that expected for Tn916. Tetracycline resistance could be transferred by conjugation to plasmid-free Lactococcus and Enterococcus strains. However, no plasmid DNA was detected among the transconjugants while both tet(M) and transposon-related sequences were amplified by PCR. This suggested that only the transposon was mobilized.     

Phenotypic and genotypic comparisons reveal a broad distribution and heterogeneity of hemolysin BL genes among Bacillus cereus isolates

The presence of hemolysin BL (HBL; components L(2), L(1), and B)-encoding genes (hblC, hblD, and hblA) from 339 Bacillus cereus strains isolated in Thailand was determined. PCR analysis showed that all three hbl genes were detected in 222 strains (65.5%). Two, one or no hbl genes were detected in 3 (0.9%), 6 (1.8%), and 108 (31.8%) strains, respectively. Among the 222 strains in which all three hbl genes were detected, 210 (61.9%) displayed discontinuous hemolysis (DH) characteristic of HBL producers, while 12 (3.5%) showed continuous hemolysis (CH) on sheep blood agar. Among strains in which none of the hbl genes was detected, 97 (28.6%) displayed CH while 11 (3.2%) did not show hemolytic activity. Three strains in which two hbl genes were detected showed CH. hblC was present in five of six strains where only one hbl gene was detected, and all of them (designated SS-00-15, TG-00-06, TG-00-14, F-00-25, and NR-01-49) showed DH. The HpaII restriction profiles of PCR fragments amplified from the hblC-A region in these five strains using hblC forward (FHC) and hblA reverse (RHA(2)) primers displayed heterogeneous patterns, which indicated sequence variation. Western blot analysis using polyclonal antibodies (Pab) raised against HBL components purified from strain F837/76 showed that three of the five strains produced all three components, whereas strain TG-00-06 did not give a signal for any component, and strain TG-00-14 produced B and L(1) but not L(2). The production of L(2) by these five strains was further analyzed using the Oxoid RPLA test. Three strains gave high titers (>64) whereas strains TG-00-06 and TG-00-14 showed lower titers of 16 and 32, respectively. The data show that HBL-encoding genes are widely distributed among B. cereus isolated in Thailand and there is a high degree of heterogeneity in both the genes and proteins. This is the first report of a B. cereus strain showing DH in which all three hbl genes and their proteins were not detected by both PCR primers and antibodies derived from prototype and type strains. The data also suggest that the L(2) component from strains TG-00-06 and TG-00-14 may be antigenically very different from that of most B. cereus isolates.