对虾原肌球蛋白致敏小鼠模型的构建及乳酸菌肠黏膜免疫效应对致敏性的影响

以原肌球蛋白（tropomyosin,TM）为主要过敏原,腹腔注射6 周龄BALB/c雌、雄小鼠,从第14天开始给治疗组小鼠口服灌胃长双歧杆菌婴儿亚种（Bifidobacterium longum subsp.）1.2202或芽孢乳酸菌09.712（Bacillus coagulans 09.712）,连续灌胃20 d。根据小鼠腹泻情况、过敏症状评分、血清中组胺（histamine,HIS）质量浓度、TM特异性免疫球蛋白（immunoglobulin,Ig）E水平变化构建TM致敏及益生菌治疗小鼠模型;通过氯乙酸萘酚酯酶染色、高碘酸希夫试剂特殊染色法检测各组小鼠肠道部位的肥大细胞和杯状细胞,通过免疫组化法检测各组小鼠肠道部位的CD4＋ T细胞和嗜酸性粒细胞;探究肠黏膜免疫在TM致敏及益生菌缓解TM致敏作用中的机理。结果表明：雌性小鼠更适用于构建TM致敏模型;益生菌灌胃可以通过降低血清中HIS质量浓度和IgE水平缓解肠黏膜免疫反应,减轻TM过敏症状。本研究从肠黏膜免疫反应角度探究了其在TM致敏中的作用,为食物过敏的研究提供参考。     

葡萄果渣酵素的发酵工艺优化及其理化特性

以果酒及果醋酿造过程中废弃的葡萄果渣、糖蜜为原料,利用酵母菌、乳酸菌、醋酸菌多菌种共酵过程中物质间的转化与利用从而得到营养价值更高的酵素保健饮品。在确定基本超声条件（超声频率40 kHz、超声功率50 W、超声温度30 ℃）的前提下,考察活菌接种量、初始pH值、糖添加量、超声时间对超氧化物歧化酶活力及活菌数的影响;并探究因素间交互作用的影响,利用响应面法设计4因素3水平试验,对葡萄果渣酵素发酵工艺进行优化,优化后得到初始pH 5.0、糖添加量8%、活菌接种量12%、超声时间60 min,并将该条件用于5 L罐的工艺放大,对补料方式进行改进,发酵终点时pH值为3.5,可滴定酸度为2.89%,总酚质量浓度为5.5 mg/mL。检测得到功能性成分如原花青素质量浓度为3.78 mg/mL,SOD活力达到988 U/mL。采用总抗氧化能力试剂盒测定得到该酵素对2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)阳离子自由基清除能力（以Trolox当量计）为（44.5±2.5）μg/g。经检测,该葡萄果渣酵素含有丰富的酒石酸、苹果酸、琥珀酸、柠檬酸,发酵过程中散发葡萄特有的醇、酚、萜类香气,葡萄果渣酵素香气独特且绵密。得到的葡萄果渣酵素上清液色泽透明亮丽,香气绵柔,口感、滋味评价均较高。     

新疆喀什地区维吾尔族母乳源双歧杆菌分离筛选及其益生特性分析

采用多重聚合酶链式反应指纹图谱结合16S rRNA基因测序技术对新疆喀什地区维吾尔族母乳分离的双歧杆菌进行鉴定和遗传差异分析,并检测常规生理生化和糖代谢表型特征,同时测试菌株对6 种常见病原菌和3 种母乳源条件致病菌的抑菌性能和对胃肠液的耐受性。结果显示,15 份母乳样品中共分离15 株双歧杆菌,测序结果将菌株归属于3 个种以及2 个亚种,包括8 株假小链双歧杆菌（Bifidobacterium pseudocatenulatum）、2 株短双歧杆菌（B. breve）、2 株长双歧杆菌长亚种（B. longum subsp. longum）和3 株长双歧杆菌婴儿亚种（B. longum subsp. infantis）。抑菌实验表明,15 株测试菌株中,隶属于B. pseudocatenulatum的4 株菌MY92、MY75-1、MY72、MY81的抑菌谱更广,抑菌能力更强;胃肠液耐受性实验表明菌株MY92无论在模拟胃液还是模拟肠液中存活率均最高,分别达到20.37%和0.302%。基于以上描述特性,MY92作为一株有效的益生菌株,具有潜在的利用价值,为后期进一步作为防止婴幼儿腹泻辅助制剂的开发提供参考。     

Enhanced phagocytosis of Aggregatibacter actinomycetemcomitans cells by macrophages activated by a probiotic Lactobacillus strain

The activation of phagocytosis is one important approach to clearing pathogenic cells in a host. This study evaluated the ability of probiotic lactobacilli to induce phagocytic activity as well as the clearance of a periodontal pathogen, Aggregatibacter actinomycetemcomitans. First, the activation of phagocytosis was found by using lyophilized dead cells. Probiotic Lactobacillus strains significantly enhanced the phagocytic activity of macrophage cells, indicating that the probiotic lactobacilli have a remarkable ability to stimulate the macrophages. Essentially, 3 Lactobacillus strains tested did not have any critical toxic effect on the murine macrophage, and Lactobacillus johnsonii NBRC 13952 showed the least cytotoxic effect on the RAW264.7 macrophages. The expression of classically activated macrophage markers, IL-1β, and cluster of differentiation 80 increased by L. johnsonii NBRC 13952; however, there was no significant difference for IL-18. The highest phagocytic activity by macrophages was found in a condition in which the macrophage activated by L. johnsonii NBRC 13952 functions to kill the cells of A. actinomycetemcomitans. Correlating with the result, a high amount of hypodiploid DNA (SubG1) was detected from the macrophage cells stimulated by L. johnsonii NBRC 13952. Taken together, the results suggest that macrophages activated by the Lactobacillus strain can facilitate the phagocytosis of A. actinomycetemcomitans cells by linking with enhanced apoptotic activities. In conclusion, L. johnsonii NBRC 13952 has a certain role in activating the RAW264.7 macrophages, thereby counteracting the infection of A. actinomycetemcomitans.     

Development of an Innovative Nutraceutical Fermented Beverage from Herbal Mate (Ilex paraguariensis A.St.-Hil.) Extract

Herbal mate (Ilex paraguariensis A.St.-Hil.) leaves are traditionally used for their stimulant, antioxidant, antimicrobial, and diuretic activity, presenting as principal components polyphenolic compounds. The aim of this work was to develop an innovative, non-dairy, functional, probiotic, fermented beverage using herbal mate extract as a natural ingredient which would also be hypocholesterolemic and hepatoprotective. Among different strains used, Lactobacillus acidophilus was selected as the best for fermentation. The addition of honey positively affected the development of L. acidophilus and the formulated beverage maintained microbial stability during shelf life. Key ingredients in the extract included xanthines, polyphenols and other antioxidants with potential health benefits for the consumer. Caffeine levels and antioxidant activity were also studied. Acceptable levels of caffeine and large antioxidant capacity were observed for the formulation when compared to other antioxidant beverages. An advantage of this product is the compliance to organic claims, while providing caffeine, other phyto-stimulants and antioxidant compounds without the addition of synthetic components or preservatives in the formulation. Sensorial analysis demonstrated that the beverage had good consumer acceptance in comparison to two other similar commercial beverages. Therefore, this beverage could be used as a new, non-dairy vehicle for probiotic consumption, especially by vegetarians and lactose intolerant consumers. It is expected that such a product will have good market potential in an era of functional foods.     

Probiotic and Technological Properties of Facultatively Heterofermentative Lactobacilli from Greek Traditional Cheeses

Nineteen isolates of facultatively heterofermentative lactobacilli from Feta, Graviera, and Kasseri cheeses were identified by SDS-PAGE of whole-cell proteins as L. paracasei subsp. paracasei (12 strains) and L. plantarum (7 strains) and differentiated at strain level by RAPD-PCR. Properties of technological interest, such as acidification ability, proteolytic activity, and enzyme activities, were also studied. The test strains exhibited a low acidification activity, with significant interstrain differences after growth in milk for 24 h. They were also characterized by different casein breakdown ability, with around 50% of them accumulating amino acids at low amounts in the milk. Lactobacilli isolates differed in respect of enzyme activities, with β-galactosidase being the strongest activity found. Their probiotic potential was evaluated with in vitro studies on the resistance to low pH, bile salts, and pancreatin. The isolates from Feta showed a better survival than those from Kasseri and Graviera at low pH and viable cells were detected even after 3 h at pH 2.0. All strains tolerated bile salts at 0.3% and retained viability in the presence of pancreatin at 0.1%. Different patterns of antibacterial activities were recorded. The strains inhibited preferentially LAB species and some of them clostridia, E. coli O44 and B. cereus. Distinguished strains are promising probiotic candidates as adjuncts and deserve further studies.     

Characterization and Technological Properties of Lactic Acid Bacteria in the Production of “Sorghurt,” a Cereal-Based Product

A total of 57 predominant LAB strains isolated from cassava and maize grains fermentation processes for fufu and ogi were identified using phenotypic and genomic fingerprinting methods such as rep-PCR and ARDRA. They were divided into facultatively heterofermentative rods (26.3%), obligately heterofermentative rods (31.6%), and tetrad forming homofermentative cocci (42.1%). Selected strains were further identified by sequencing the 16S rDNA gene. Technological studies such as acidification, hydrogen peroxide production, starch hydrolysis, enzymatic activities, degradation of oligosaccharides, and in vitro adherence properties were carried out. Lactobacillus plantarum strains demonstrated better and rapid acid production capability, followed by the Pediococcus strains, while L. fermentum strains exhibited slower acid production. Hydrogen peroxide production was observed among the LAB groups. The test strains utilized the indigestible sugars raffinose and stachyose. Only L. pentosus demonstrated high amylase activity comparable to that of L. amylovorus DSM 20531. Lactobacillus plantarum and L. fermentum strains showed high β-glucosidase activity. Six strains were selected as starter cultures. The strains were tolerant to acidic pH levels and bile salt. The yoghurt-like “sorghurt” produced using the selected starter cultures had a final pH of less than pH 4.0, and a viable count of less than 5.5 Log₁₀cfu/mL at the end of a 24 h fermentation period. The samples were generally acceptable to the taste panelists. The starter organisms demonstrated varying degree of adherence to HT29 MTX cell line. Therefore, employing functionally defined LAB strains may be one practical approach for incorporating health-promoting features into appropriate food products suitable for targeted population.     

Enhancement of antimicrobial and antimutagenic activities of Korean barberry (Berberis koreana Palib.) by the combined process of high‐pressure extraction with probiotic fermentation

BACKGROUND: To evaluate the combined effects of high pressure extraction (HPE) and probiotic fermentation on the antimicrobial and antimutagenic activities, Berberis koreana was subjected to 500 MPa for 30 min and then fermented with Bifidobacterium longum B6 (HPE‐BLF) and Lactobacillus paracasei (HPE‐LPF) at 37 °C for 6 days. RESULTS: The phenol content was significantly increased to 228 mg GAE g^?1 by the HPE compared to the conventional extraction (CE, 188 mg GAE g^?1). The HPE‐BLF and HPE‐LPF showed the highest antimicrobial activity (MIC < 4 mg mL^?1) against β‐lactam antibiotic sensitive and resistant Staphylococcus aureus. No significant mutagenic effect was observed for CE, HPE, HPE‐BLF, and HPE‐LPF extracts. The highest antimutagenic activities against frame‐shift mutant Salmonella typhimurium were observed at the HPE‐LPF (82%), followed by the HPE‐BLF (77%). CONCLUSION: The combined HPE and fermentation process could be used as an alternative extraction method for improving the extraction efficacy of medicinal plants. The results will provide pharmaceutically useful information and potential direction for finding new drug sources from medicinal plants. Copyright © 2010 Society of Chemical Industry     

青藏高原牦牛酸奶中具有抗氧化活性乳酸菌的体内外益生特性

目的:筛选出性状优良的具有抗氧化性的益生乳酸菌并研究其对D-半乳糖致衰老模型大鼠的抗氧化作用。方法:以从青藏高原牦牛酸奶中初步筛选的3株具高抗氧化活性乳酸菌为研究对象,通过耐受模拟胃肠道、耐胆盐和疏水能力实验评价3株乳酸菌的体外益生特性,并将体外益生特性优良的菌株饲喂D-半乳糖致衰老模型大鼠。通过检测各处理组大鼠的脏器指数,肝脏、脑组织和血清中的谷胱甘肽过氧化物酶(glutathion peroxidase,GSH-Px)、总超氧化物歧化酶(total superoxide dismutase,T-SoD)活力、总抗氧化能力(total antioxidant capacity,T-AoC)以及丙二醛(malondialdehyde,MDA)含量来评价乳酸菌的体内抗氧化益生特性。结果:植物乳杆菌(Lactobacillus plantarum)XM5具有较强的耐模拟胃肠液和耐胆盐能力,同时具有较强的疏水能力。植物乳杆菌XM5的摄入能有效缓解D-半乳糖对大鼠脑组织的氧化损伤;使衰老模型大鼠肝脏中的GSH-Px活力,血清中的GSHPx、T-SoD活力和T-AoC显著提高(P0.05);同时使肝脏、脑组织和血清中的MDA含量显著降低(P0.05)。结论:植物乳杆菌XM5是一株性能优良的抗氧化益生乳酸菌,具有潜在的应用价值。