A novel immunostimulating aspect of Lactobacillus gasseri: induction of "Gasserokine" as chemoattractants for macrophages

The chemotactic activity of the culture supernatants from 14 strains of Lactobacillus acidophilus and L. gasseri was examined for murine macrophages. Significant macrophage chemotactic activity was observed in three strains of L. acidophilus and all strains of L. gasseri. The highest activity was observed in the supernatant (1131-sup) from 24-h cultures of L. gasseri JCM1131T. The chemotactic factor from 1131-sup, designated as "Gasserokine", was purified by the C18 reverse phase and ion-exchange chromatography. The purity of Gasserokine was checked by HPLC with the reverse-phase mode. The chemotactic activity of Gasserokine was also observed for human monocytes. The macrophage chemotaxis induced by L. gasseri JCM1131T culture supernatants was discovered to be a new biological function exerted by probiotic lactic acid bacteria. Therefore, the activity is expected to be used for one of the functional parameters in the immunomodulating properties of probiotic lactic acid bacteria.     

核桃粕发酵乳菌种筛选及发酵条件优化研究

该试验对乳酸菌发酵核桃粕乳的不同菌株进行筛选,通过pH值、酸度及感官评定分析,从9株乳酸菌中筛选出嗜酸乳杆菌(Lactobacillus acidophilus)和植物乳杆菌(Lactobacillus plantarum)两株优良菌种。研究两株乳酸菌的复配比例,并与传统发酵剂发酵的核桃粕乳进行品质对比。结果表明,以嗜酸乳杆菌:植物乳杆菌(1∶1)接种发酵后得到的发酵核桃粕发酵乳综合品质最佳,其感官评分90分,氨基酸态氮含量57.0 mg/L,活菌总数7.35×107 CFU/mL,经发酵后的营养价值明显优于传统发酵剂发酵的核桃粕乳。     

肉制品发酵剂菌种筛选及发酵工艺条件优化

为获得优良的肉制品发酵剂,将酸马奶酒中分离所得4株乳酸菌进行培养。以标准的植物乳杆菌为对照,通过生化特性分析、耐盐性、耐亚硝酸盐性、产酸能力等试验对其进行优势菌种筛选,最终筛选出一株发酵性能较好的乳酸菌Lactobacillus5(BL5)。以接菌量、BL5和标准肉葡萄球菌Staphylococcus(Stl)配比、葡萄糖添加量作为影响发酵的三个因素,测定pH达到5.3左右的发酵时间和色泽。结果表明:当接菌量1×106cfu/g,BL5︰Stl=2︰1,葡萄糖添加量2%时,发酵时间较短为8 h,pH为5.2±0.1,L*,a*值都较大,发酵肉的色泽较好。故将其作为该发酵剂的最佳发酵条件。     

广西地区青年人肠道乳酸菌的分离及其发酵酸奶品质的评价

采用传统的可培养方法对广西青年人肠道乳酸菌进行了分离与纯化,通过16S rRNA序列分析对菌株进行了鉴定,并将筛选菌株作为酸奶发酵剂应用。结果表明,分离到的23株乳酸菌中被鉴定为发酵乳杆菌（Lactobacillus fermentum）11株,唾液乳杆菌（Lactobacillus salivarius）6株,格氏乳杆菌（Lactobacillus gasseri）2株,副干酪乳杆菌（Lactobacillus paracasei）1株,融合魏斯氏菌（Weissella confuse）2株和口乳杆菌（Lactobacillus oris）1株。筛选乳酸菌制作的酸奶之间芳香类物质差异并不显著（P＞0.05）,而酸奶样品间酸味、后味A和丰度差异较大。进一步分析表明,副干酪乳杆菌HBUAS54287、唾液乳杆菌HBUAS54223与HBUAS54248、格氏乳杆菌HBUAS54233、发酵乳杆菌HBUAS54238与HBUAS54318乳酸产生能力较强,可进一步筛选用于酸奶复合发酵剂的开发。     

Metabolic and biochemical responses of probiotic bacteria to oxygen

The interaction between oxygen and probiotic bacteria was studied by growing Lactobacillus acidophilus and Bifidobacterium spp. in 0, 5, 10, 15, and 21% oxygen in a hypoxic glove box. The metabolic responses of each probiotic strain in the different oxygen environments were monitored by measuring the levels of lactic acid and determining the lactate-to-acetate ratio. Biochemical changes induced by oxygen were examined by monitoring the specific activities of NADH oxidase, NADH peroxidase, and superoxide dismutase. In addition, the ability to decompose hydrogen peroxide and the sensitivity of each strain to hydrogen peroxide was also determined. With an increase in oxygen percentage, levels of lactic acid in L. acidophilus strains decreased, whereas the lactate-to-acetate ratio reduced in all the bifidobacteria tested. At 21% oxygen, the specific activities of NADH oxidase and NADH peroxidase, and the hydrogen peroxide decomposing ability of five probiotic strains was significantly higher than at 0% oxygen. The sensitivity of the probiotic strains to hydrogen peroxide however, remained unaffected in all the different oxygen percentages. Superoxide dismutase levels did not reveal any conclusive trend. In both L. acidophilus and Bifidobacterium spp., NADH oxidase and NADH peroxidase functioned optimally at pH 5. Growth in the various oxygen environments did not change this optimum pH.     

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。     

Effects of Lactic Acid Bacteria and Fermented Milks on Eicosanoid Production by Intestinal Epithelial Cells

ABSTRACT: Fermented milk products produced with probiotic lactic acid bacteria (LAB) have attracted interest due to their potential health benefits. Probiotic bacteria have a range of immunomodulatory activity, interacting with a variety of cell types in the immune system. Interactions with intestinal epithelial cells (IEC) are an avenue through which probiotics and their fermented milks can influence production of key immunoregulatory molecules, including cytokines and eicosanoids. The eicosanoids, which include the prostaglandins (PGs), are lipid mediators implicated in both acute and chronic inflammatory processes. The primary objective of this study was to determine the ability of probiotic LAB and their ferments to interact with IEC and influence their eicosanoid production. Effects of LAB and their milk ferments on prostaglandin E₂ (PGE₂) and prostaglandin F_2α (PGF_2α) production by human IEC lines were determined using a competitive enzyme immunoassay. LAB alone did not alter interleukin (IL)-1β-induced prostaglandin production by IEC. However, milk fermented with Lac-tobacillus (L.) rhamnosus strain R0011 significantly suppressed IL-1β-induced levels of PGE₂ and PGF_2α, an effect which was counteracted by the addition of strain R0011. Milk ferments prepared withL. acidophilus strain R0052 were less effective in down-regulation of PG production by IEC. Naltrexone, an opioid receptor antagonist, blocked the suppressive effects of L. rhamnosus R0011 milk ferments on PGF_2α production by IEC, suggesting that the bioactivity the ferments is opioid receptor-mediated. These findings support immunomodulatory potential of fermented food components through interactions with intestinal epithelial cells.     

酵母菌乳酸菌共发酵对荔枝汁品质的影响

本研究以鲜榨荔枝汁为原料,分别对其进行干酪乳杆菌及三种酵母菌(BO213、D254、EC1118)的复合发酵,探索发酵过程中乳酸菌活菌数、pH、可溶性固形物、糖组分、有机酸、DPPH?清除能力以及感官指标等变化。结果表明：发酵后荔枝汁中的乳酸菌活菌数均高于7 lg (cfu/mL),均具有一定的益生功能。复合发酵不仅能促进干酪乳杆菌的生长,还能降低荔枝汁的糖度,且酒精度低于0.50%,符合无醇饮料的标准。随着发酵时间的延长,各组的糖含量均呈现下降的趋势。发酵24 h后干酪乳杆菌单独发酵组的乳酸含量显著低于复合发酵组,从高到低依次为：D254+干酪组(6.62 g/L)>Ec1118+干酪组(6.55 g/L)>BO213+干酪组(2.35 g/L)>干酪组(0.94 g/L)。D254+干酪组、Ec1118+干酪组发酵24 h的荔枝汁体外抗氧化能力(DPPH?清除能力)相比单独发酵组更具优势。发酵后的荔枝汁香气更佳,且复合发酵优于单独发酵,其中BO213+干酪组的香气最好,Ec1118+干酪组次之。综上,通过Ec1118+干酪乳杆菌可制得一款风味良好、营养丰富的发酵无醇益生荔枝饮料。     

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.     

益生菌对发酵菠萝果汁综合品质的影响

菠萝果汁分别接种干酪乳杆菌、植物乳杆菌、嗜酸乳杆菌和肠膜明串珠菌进行发酵,研究菠萝果汁发酵过程中理化品质和感官品质的变化规律。结果表明,4种乳酸菌在菠萝果汁中生长良好,发酵24 h,植物乳杆菌和肠膜明串珠菌的活菌数对数值均已超过8.3;经乳酸菌发酵后,菠萝果汁的pH、维生素C（VC）含量均显著降低,而总酸、可溶性固形物、总酚、1,1-二苯基-2-三硝基苯肼（DPPH）自由基清除率显著升高。其中,植物乳杆菌具有较强的产酸能力,肠膜明串珠菌能够延缓VC的流失。在对感官品质影响方面,4种发酵菠萝果汁之间存在差异,经肠膜明串株菌发酵的菠萝果汁感官评价较优（79.79分）,而经嗜酸乳杆菌能够更好地保留菠萝果汁特有的挥发性香气成分。研究结果为益生菌菠萝果汁产品的开发提供参考依据和技术支持。     

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.     

乳酸菌在柿子汁发酵中的应用研究

以柿子汁为原料，分别经乳酸菌属中的保加利亚乳酸杆菌，嗜酸乳杆菌，嗜热链球菌单个或两个混合发酵而制成柿子汁乳酸菌发酵饮料。通过对接种量，产酸量，发酵时间，发酵温度和风味等方面进行试验，从而确定了最佳的优良菌种及其在柿子汁中最佳的发酵条件。     

Ex vivo effects of lactobacilli, streptococci, and bifidobacteria ingestion on cytokine and nitric oxide production in a murine model

Increasing numbers of functional foods and pharmaceutical preparations are being promoted with health claims based on the potential probiotic characteristics of lactic acid bacteria and on their capacity for stimulating the host immune system. However, the specific immune effects of oral administration of these microbes still remains undefined. In this study, we tested the hypothesis that production of immunologic mediators by leukocytes in mice is affected by orally administered lactic acid bacteria. The specific objectives of this study were to evaluate the effects of exposure to eight different lactic acid bacteria in mice on ex vivo cytokine and nitric oxide production in leukocyte cultures. Mice were gavaged with 1 X 10(9) viable bacteria and peritoneal, Peyer's patch and splenic leukocytes were isolated 8 h later. These were cultured for 2 or 5 days in the presence or absence of mitogens and then interleukin (IL)-6, IL-12, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and nitric oxide production was measured. The results revealed that Lactobacillus acidophilus and L. casei potentiated IL-6 and IL-12 production by peritoneal cells whereas L. acidophilus upregulated IFN-gamma and nitric oxide. In contrast, L. helveticus, L. gasseri, L. reuteri, and Bifidobacterium attenuated the production of IL-6, IFN-gamma, and nitric oxide by peritoneal cells. TNF-alpha was not detectable in peritoneal cultures. None of the bacteria altered ex vivo production of cytokines or nitric oxide by Peyer's patch or spleen cell cultures. Taken together, the results suggest that prior oral exposure to lactic acid bacteria could differentially potentiate or attenuate subsequent cytokine and nitric oxide production by peritoneal cells.     

Preparation of a Lactobacillus plantarum starter culture for cucumber fermentations that can meet kosher guidelines

A method is described for growth of a Lactobacillus plantarum starter culture in jars of commercially available pasteurized fresh-pack kosher dill cucumbers so that jars can be used to inoculate commercial scale cucumber fermentation tanks. A procedure is also described to transfer lactic acid bacteria from frozen storage in MRS broth into cucumber juice and commercial jars of kosher dill cucumbers so that a selected strain of lactic acid bacteria can be kosher certified for commercial fermentations in processing plants that operate under kosher certification. The strain of L. plantarum used in these experiments grew to maximum cell numbers in 4 d at 20 to 25 °C and then maintained viable cell numbers for 2 wk at >10(8) CFU/mL so the culture was suitable for inoculation of fermentation tanks. Refrigeration of jars of culture after they grow to maximum numbers minimizes die-off of cells sufficiently so that a pure culture can be maintained by aseptically transferring brine containing viable bacteria to a new pH-adjusted jar only once every 4 mo. PRACTICAL APPLICATION: This report describes a method to prepare a lactic acid bacteria starter culture suitable for kosher vegetable fermentations.     

Species-specific identification of commercial probiotic strains

Products containing probiotic bacteria are gaining popularity, increasing the importance of their accurate speciation. Unfortunately, studies have suggested that improper labeling of probiotic species is common in commercial products. Species identification of a bank of commercial probiotic strains was attempted using partial 16S rDNA sequencing, carbohydrate fermentation analysis, and cellular fatty acid methyl ester analysis. Results from partial 16S rDNA sequencing indicated discrepancies between species designations for 26 out of 58 strains tested, including two ATCC Lactobacillus strains. When considering only the commercial strains obtained directly from the manufacturers, 14 of 29 strains carried species designations different from those obtained by partial 16S rDNA sequencing. Strains from six commercial products were species not listed on the label. The discrepancies mainly occurred in Lactobacillus acidophilus and Lactobacillus casei groups. Carbohydrate fermentation analysis was not sensitive enough to identify species within the L. acidophilus group. Fatty acid methyl ester analysis was found to be variable and inaccurate and is not recommended to identify probiotic lactobacilli.     

4株乳酸菌对模拟胃肠环境的耐受性及生长特性研究

对4株来源于自然发酵肉制品中的乳酸菌进行耐酸耐胆盐性能的测定,选取在pH3.0和含0.3 g/100 mL胆盐的MRS培养基中存活率均超过50%的2株菌（M12、M23）,进一步研究其在模拟胃肠液中的耐受性及MRS培养基中的生长特性。结果表明,这2株乳酸菌对人工模拟胃肠道环境有较好的耐受性,经过人工模拟胃液处理2 h后,存活率均＞50%,经人工模拟肠液处理10 h后仍可存活;这2株菌具有较强的增殖及产酸能力,24 h内均已进入稳定生长期,随着菌体数目的增加,培养基pH值快速降低,在肉制品的发酵温度范围内均能较好地生长。因此,这2株菌可作为潜在的益生菌菌株应用于肉制品的发酵。     

Improved sauerkraut production with probiotic strain Lactobacillus plantarum L4 and Leuconostoc mesenteroides LMG 7954

Probiotic strain Lactobacillus plantarum L4 and strain Leuconostoc mesenteroides LMG 7954 were applied for the controlled fermentation of cabbage heads. The parameters of the controlled and spontaneous fermentations, including antimicrobial effect of cabbage brines obtained at the end of both fermentations, were monitored. To check out the influence of starter culture strains, 10 randomly chosen lactic acid bacteria, isolated at the end of controlled cabbage heads fermentation were identified by API 50 CHL test, and the presence of the probiotic culture was confirmed by Pulsed Field Gel Electrophoresis. The starter cultures applied for cabbage heads fermentation allowed lowering of NaCl concentrations from 4.0% to 2.5% (w/v), considerably accelerated fermentation process by 14 d, and improved the product quality. The produced sauerkraut heads are considered probiotic product as viable probiotic cells count in final product was higher than 10(6) colony-forming units (CFU) per gram of product. PRACTICAL APPLICATION: The results of this research could be applied in the production of fermented cabbage heads with added functional (probiotic) value and with lower NaCl concentration with expected shortened fermentation time. This could not only be of economic but also of ecological importance.     

冷应激及包埋对乳酸菌冻干发酵剂的影响

将嗜热链球菌、保加利亚乳杆菌、嗜酸乳杆菌按3∶2∶1比例制备乳酸菌冻干发酵剂.比较不同温度冷应激处理对乳酸菌冻干存活率的影响,结果表明,20℃,处理4h时,冻干存活率为96.3％.选择发酵的固定化细胞制作条件:海藻酸钠2.25g/L、CaCl2 4.0g/L.包埋后乳酸菌冻干存活率为95.6％.     

乳酸菌发酵大豆糖蜜生产乳酸及糖代谢变化

选用德氏乳杆菌保加利亚亚种KLDS1.8501、嗜酸乳杆菌KLDS1.0327、嗜酸乳杆菌ATCC11975、植物乳杆菌植物亚种CICC23168、干酪乳杆菌ATCC393、植物乳杆菌NAU322分别接种于大豆糖蜜,用高效液相色谱法测定乳酸的产生以及碳水化合物的利用情况,分析不同乳酸菌发酵大豆糖蜜生产乳酸能力及糖代谢能力。结果表明,在15 °Brix大豆糖蜜中,37?℃、pH?6.0条件下发酵24?h,植物乳杆菌植物亚种CICC23168的活细胞数达到6.66×109?CFU/mL,乳酸产生量为12.18?g/L,总糖消耗量为22.48?g/L,与其他菌株相比有明显优势。因此,植物乳杆菌植物亚种CICC23168是能利用大豆糖蜜发酵产乳酸的潜力菌株。     

不同乳杆菌强化发酵红薯叶及其酸菜品质的研究

为筛选发酵红薯叶最适宜的乳酸菌菌种,以植物乳杆菌、嗜酸乳杆菌、鼠李糖乳杆菌和混合乳酸菌强化发酵红薯叶,结果表明,乳酸菌强化发酵酸菜的pH下降快、乳酸含量高、亚硝酸盐含量低,可溶性多糖含量高。其中,混菌发酵乳酸含量最高(6.19mg/g,P<0.05),亚硝酸盐含量最低(0.29μg/mL,P<0.05),多糖含量(22.90mg/g)与单菌发酵无显著差异;混菌发酵红薯叶硬度、咀嚼性和紧实度均最大;混菌发酵有24种氨基酸,比自然发酵多4种,呈鲜味氨基酸最丰富,必需氨基酸最齐全且含量最高(1 561.72μg/g,P<0.05)。