乳杆菌DM9811挥发性脂肪酸组分对白色念珠菌、金黄色葡萄球菌的抑制

目的：探讨乳杆菌DM9811发酵液的挥发性脂肪酸对金黄色葡萄球菌、白色念珠菌抑制作用。方法：采用普通细菌计数法。结果：乳杆菌DM9811发酵液中挥发性脂肪酸对金黄色葡萄球菌、白色念珠菌具有抑制作用,以对白色念珠菌抑制作用最强,金黄色葡萄球菌抑制次之,两者都呈现剂量效应关系。结论：乳杆菌DM9811发酵液中挥发性脂肪酸对细菌、真菌都有明显的抑制作用。     

乳杆菌对外阴道白假丝酵母菌病治疗的分子机制

目的通过应用乳杆菌代谢产物提取物治疗外阴道白假丝酵母菌的效果,探讨乳杆菌治疗白假丝酵母菌病的可能分子机制。方法选择符合外阴道白假丝酵母菌病患者57例,随机分为2组,组应用乳杆菌代谢产物提取物治疗7d(试验组);另一组采用常规治疗(对照组),观察治疗前后阴道pH的变化,白假丝酵母菌、乳杆菌的变化及临床治疗效果的变化。结果采用乳杆菌代谢产物提取物治疗后阴道pH值恢复正常,白假丝酵母菌降低,乳杆菌变化不明显,临床治疗效果明显。结论乳杆菌治疗白假丝酵母菌病的分子机制可能是乳杆菌持续产生的乙酸、9-十六碳烯酸、十六烷酸、9,12-十八碳二烯酸和9-十八碳烯酸作用的结果。     

乳杆菌DM9811发酵液提取物中RNA组分对人结肠癌细胞系HT-29增殖影响及分子机制研究

目的观察乳杆菌DM9811发酵液提取物中RNA组分对人结肠癌细胞系HT-29增殖的影响,探讨其对肠道肿瘤细胞的作用及其分子机制,为阐明乳杆菌与宿主相互作用规律的分子机制奠定基础。方法应用MTT方法研究不同时间不同浓度RNA组分对HT-29增殖的影响,应用流式细胞术、RT-PCR研究RNA组分对HT-29细胞周期的影响。结果乳杆菌DM9811发酵液中RNA组分能抑制HT-29细胞增殖,并呈现出时间-剂量依赖性;RNA组分作用于HT-29细胞24 h、48 h时,细胞周期G0/G1期所占比例明显上升,S期所占比例明显降低(P0.01),细胞周期调控因子CDK6、p27Kip1、p53的表达升高,CDK2、CDK4、PCNA的表达降低。结论乳杆菌DM9811代谢产物RNA在体外具有抑制癌细胞周期活性。     

发酵无花果香料的挥发性成分分析

利用微生物发酵无花果开发特色香料,并采用同时蒸馏萃取装置收集挥发性成分并用气相色谱一质谱仪对生物技术制备的无花果香料挥发性成分进行分离和鉴定,经毛细管色谱分离出47种组分,确认了其中的45种成分,并用面积归一化法测定了各种成分的百分含量,其主要成分为：9,12-十八碳二烯酸乙酯(27．34％)、十六酸乙酯(23．99％)、邻苯二甲酸二丁酯(6．18％)、邻苯二甲酸二异丁酯(5．52％)、9,12-十八碳二烯酸甲酯(4．72％)、十六酸甲酯(4．67％)、9,12,15-十八碳三烯酸乙酯(4．48％)、9-十八碳烯酸乙酯(3．80％)、糠醛(2．53％)、9,12,15-十八碳三烯酸甲酯(1．85％)、十八酸乙酯(1．42％)、9-十八碳烯酸甲酯(1．26％)等。     

瑞香狼毒茎叶化学成分研究

新鲜瑞香狼毒(Stellera chamaejasme)茎叶经正已烷提取,脱腊后进行GC-MS-DS联用分析,鉴定出20个化合物。主要成分为正十八烷;正十九烷;2,6,10,14 — 四甲基十六烷;十六烷酸;十六烷酸甲酯;十六烷酸乙酯;9,12,15-十八碳三烯酸甲酯;9,12-+八碳二烯酸;9,12,15-十八碳三烯酸和十八烷酸。     

乳杆菌DM9811代谢产物脂肪酸组分对义齿性口炎患者口腔中白假丝酵母菌的抑制

目的从调整口腔菌群平衡的角度出发,观察乳杆菌DM9811代谢产物脂肪酸组分对义齿性口炎患者口腔菌群的影响。方法（1）体外试验：配制不同浓度乳杆菌DM9811代谢产物脂肪酸组分的沙包液体培养基,分别接种10^8CFU／ml白假丝酵母菌菌液,用CFU活菌记数法计数。（2）临床试验：符合义齿性口炎病例40例,随机分为试验组20例和对照组20例（阴性对照组和阳性对照组各10例）。分别给予乳杆菌DM9811代谢产物脂肪酸组分、纯净水、洗必泰含漱液。观察菌群变化。结果（1）体外试验：乳杆菌DM9811代谢产物脂肪酸组分在浓度为1．6、0．8和0．4mg／ml时对白假丝酵母菌的抑制作用分别为100％、98．11％和62．72％。（2）临床试验：试验组在治疗后口腔黏膜白假丝酵母菌数量明显降低,口腔链球菌数量升高,较治疗前差异均有显著性（P〈0．01）;停药后第21天较治疗7d差异无显著性（P〉0.05）。结论乳杆菌DM9811代谢产物脂肪酸组分对口腔白假丝酵母菌有明显的抑杀作用;对口腔链球菌有促进作用,可能有调整口腔菌群平衡的作用。     

大麦虫不同虫态甲醇粗提物化学成分的分析和比较

为了探寻大麦虫Zophobas morio变态发育过程中化学成分的变化, 本研究利用薄层色谱技术对大麦虫的3个虫态(幼虫、 蛹和成虫)甲醇粗提物中的化学成分进行分析和比较, 利用柱色谱技术、 核磁共振氢谱技术（¹H MR）和气相色谱-质谱联用技术（GC-MS）对蛹甲醇粗提物进行重点分析。结果表明： 虫蛹甲醇粗提物中含有幼虫和成虫甲醇粗提物中不存在的化学成分。对虫蛹甲醇粗提物进行针对性的结构研究, 共鉴定出10个脂肪酸。其中, 存在于大麦虫虫蛹中的10-十六碳酮酸、 10-十八碳酮酸为首次从自然界昆虫中获得; (8E, 11E)-8, 11-十八碳烯酸、 (9Z, 12Z)-9, 12-十八碳烯酸、 (9Z, 12Z, 15Z)-9, 12, 15-十八碳烯酸、 8-(3-辛基-2-环氧乙烷基)辛酸和8-(2-辛基环丙烷基)辛酸为首次从大麦虫中发现。以上研究结果为针对大麦虫不同生物阶段的开发利用提供新的理论依据。     

乳杆菌DM9811代谢产物脂肪酸组分对义齿性口炎的临床应用研究

目的从口腔菌群平衡的角度观察乳杆菌DM9811代谢产物脂肪酸组分对义齿性口炎患者口腔细菌的影响。方法采取自身前后对照,24例义齿性口炎患者给予乳杆菌DM9811代谢产物脂肪酸组分制成的含漱液,每次20 ml,每日3次漱口。分别于用药前,用药后第7天、14天、28天以及停药后第60天采集标本培养,记录活菌数。结果24例患者中18例为有效病例。用药后7 d口腔白假丝酵母菌数量明显降低,口腔链球菌数量升高,较治疗前差异均有显著性（P〈0.01）,乳杆菌治疗前后差异无显著性（P〉0.05）;用药14 d、28 d及停药后60 d时,口腔内白假丝酵母菌、口腔链球菌、乳杆菌数量较7 d时差异均无显著性（P〉0.05）。结论乳杆菌DM9811代谢产物脂肪酸组分治疗义齿性口炎7 d时致病菌和优势菌即可达到相对平衡。乳杆菌DM9811代谢产物脂肪酸组分治疗义齿性口炎不会造成口腔菌群的失衡。     

乳杆菌DM9811发酵液提取物中RNA组分对水泡性口炎病毒抑制作用研究

目的通过乳杆菌DM9811发酵液提取物中RNA组分对水泡性口炎病毒(VSV)抑制作用研究,探讨其在抵抗肠道病毒性感染性疾病方面的作用。方法应用50%细胞感染计量法(TCID50)、免疫荧光法、ELISA和MTT法探讨不同浓度的RNA组分对VSV的抑制作用。结果 RNA组分200μg/mL浓度时与对照比较,RNA组分具有竞争性抑制VSV感染作用,细胞存活率为(90.9±3.67)%,并具有阻断VSV侵入细胞作用,细胞存活率为(96.6±1.47)%。但RNA组分对病毒生物合成的抑制作用不明显。此外RNA组分具有诱导BALB/c小鼠脾细胞产生IFN-α作用,并呈现一定的剂量依赖关系。结论乳杆菌DM9811发酵液提取物中RNA组分对VSV具有明显抑制作用。     

乳杆菌DM9811代谢产物中的RNA组分对宫颈癌细胞cAMP/cGMP影响

目的通过分析乳杆菌DM9811代谢产物中的RNA组分对宫颈癌细胞cAMP/cGMP影响,探讨乳杆菌对细胞作用的分子机制。方法采用细胞与分子生物学方法。结果乳杆菌代谢产物中的RNA组分对细胞的信息分子cAMP具有重要的调节活性。当除去代谢产物中的RNA分子刺激细胞时,细胞内的cAMP水平低于纯化后的RNA组cAMP水平,低了1.75倍,cGMP水平也出现了对应关系。结论乳杆菌DM9811代谢产物中的RNA组分对细胞内信息分子cAMP/cGMP具有调节作用。     

乳酸杆菌、双歧杆菌代谢产物的气相色谱分析

用气相色谱分析乳酸杆菌、双歧杆菌提取物,结果表明乳酸杆菌提取物中乙酸的含量很高,双歧杆菌提取物挥发性脂肪酸种类较多,有乙酸、丙酸、异丁酸及丁酸等多种短链脂肪酸.分析结果表明短链脂肪酸很可能是乳酸杆菌、双歧杆菌提取物抑菌的主要活性成分.     

Selected nondigestible carbohydrates and prebiotics support the growth of probiotic fish bacteria mono-cultures in vitro

Aims:  To search for nondigestible but fermentable (NDF) carbohydrates and prebiotics with a potency to promote the growth of selected bacteria in vitro .
Methods and Results:  The growth of three reference bacteria strains Bacillus subtilis LMG 7135^T, Carnobacterium piscicola LMG 9839, Lactobacillus plantarum LMG 9211 and one candidate probiotic bacteria Lactobacillus delbrueckii subsp. lactis was investigated over a minimum period of 48 h in the presence of β -glucan, xylo-oligosaccharide, arabinoxylo-oligosaccharide, inulin, oligofructose and glucose. Besides the capability to grow on inulin and oligofructose containing media, a distinct high growth in β -glucan based substrates and a low growth in (arabino)xylooligosaccharide containing media were evident for most bacteria tested. With the exception of B. subtilis and L. plantarum , other bacteria grew equally well or even better on different substrates than on glucose. The fermentation of studied carbohydrates by these micro-organisms was dominated by the production of acetic acid as the main short chain fatty acid.
Conclusions:  Selected bacteria are able to ferment and grow on NDF and prebiotic carbohydrates but in a substrate dependent manner.
Significance and Impact of the Study:  This study delivers a first screening of which NDF or prebiotic carbohydrates are the most promising for aquaculture feed supplementations.     

Amino acid fermentation in non-starter Lactobacillus spp. isolated from cheddar cheese

Amino acid fermentation profiles of nine strains of Lactobacillus spp., initially isolated from a 3-year-old Cheddar cheese, were determined using the Biolog MT microplatetrade mark method. Eight of the isolates were able to ferment amino acids, but only when incubated in the presence of exogenously supplied alpha-ketoglutaric acid that served as an acceptor in the initial transamination step in the fermentative degradation. The range of amino acids catabolized was strain dependent. Amino acid catabolites were detected by gas chromatography mass spectrometry (GCMS) in culture supernatant fluids of a representative non-starter lactic acid bacteria isolate Lactobacillus paracasei CI6.     

Growth and end product formation of two psychrotrophic Lactobacillus spp. and Brochothrix thermosphacta ATCC 11509T at different pH values and temperatures.

Lactobacillus viridescens, Lactobacillus sp. strain 173 (homofermentative), and Brochothrix thermosphacta ATCC 11509T were studied at different pH values and temperatures in aerobic and anaerobic batch cultures. The growth rates were higher in aerobic than in anaerobic cultures. L. viridescens grew faster at pH 5.8 than at pH 6.3, whereas the opposite was true for B. thermosphacta. Lactobacillus sp. strain 173 was inhibited in air or at 8 degrees C in anaerobic culture. B. thermosphacta did not grow in anaerobic culture at pH 5.3. The following variations in growth yields were found in the different environments studied: Lactobacillus sp. strain 173, 23 to 25 g (dry weight) per mol of glucose consumed; L. viridescens, 11 to 23 g/mol; B. thermosphacta, 16 to 38 g/mol. In air, L. viridescens produced D-lactic acid, ethanol, and acetic acid, whereas no acetic acid was produced anaerobically. Acetic acid and ethanol together constituted 41 to 48% of the total product yield irrespective of pH and temperature. Lactobacillus sp. strain 173 produced a racemic mixture of D- and L-lactic acid at pH 6.3, whereas the proportion of L-lactic acid was higher than that of D-lactic acid at pH 5.3. In air, product formation of B. thermosphacta varied from a domination of L-lactic acid to increasing yields of acetoin, acetic acid, 2,3-butanediol and isovaleric acid. The effect of pH and temperature on product formation was difficult to separate from the effect of O2 availability in aerobic cultures. However, it was indicated that more 2,3-butanediol and less acetoin were produced with a decreasing temperature.(ABSTRACT TRUNCATED AT 250 WORDS)     

Growth and end product formation of two psychrotrophic Lactobacillus spp. and Brochothrix thermosphacta ATCC 11509T at different pH values and temperatures

Lactobacillus viridescens, Lactobacillus sp. strain 173 (homofermentative), and Brochothrix thermosphacta ATCC 11509T were studied at different pH values and temperatures in aerobic and anaerobic batch cultures. The growth rates were higher in aerobic than in anaerobic cultures. L. viridescens grew faster at pH 5.8 than at pH 6.3, whereas the opposite was true for B. thermosphacta. Lactobacillus sp. strain 173 was inhibited in air or at 8 degrees C in anaerobic culture. B. thermosphacta did not grow in anaerobic culture at pH 5.3. The following variations in growth yields were found in the different environments studied: Lactobacillus sp. strain 173, 23 to 25 g (dry weight) per mol of glucose consumed; L. viridescens, 11 to 23 g/mol; B. thermosphacta, 16 to 38 g/mol. In air, L. viridescens produced D-lactic acid, ethanol, and acetic acid, whereas no acetic acid was produced anaerobically. Acetic acid and ethanol together constituted 41 to 48% of the total product yield irrespective of pH and temperature. Lactobacillus sp. strain 173 produced a racemic mixture of D- and L-lactic acid at pH 6.3, whereas the proportion of L-lactic acid was higher than that of D-lactic acid at pH 5.3. In air, product formation of B. thermosphacta varied from a domination of L-lactic acid to increasing yields of acetoin, acetic acid, 2,3-butanediol and isovaleric acid. The effect of pH and temperature on product formation was difficult to separate from the effect of O2 availability in aerobic cultures. However, it was indicated that more 2,3-butanediol and less acetoin were produced with a decreasing temperature.(ABSTRACT TRUNCATED AT 250 WORDS)     

植物乳杆菌DY6主要抑菌代谢物的分析和鉴定

【背景】被广泛应用于食品和饲料等多个行业的乳酸菌已成为制作生物防腐剂的研究热点。【目的】探究抑菌性能良好的植物乳杆菌DY6的抑菌物质,为其进一步应用提供参考依据。【方法】对植物乳杆菌发酵液中抑菌物质的理化特性进行研究,采用GC-MS分析发酵上清液代谢物,通过多元统计学分析方法推测主要抑菌物质,抑菌物质通过半制备进行初步分离后用GC-MS鉴定。【结果】植物乳杆菌DY6对金黄色葡萄球菌、大肠杆菌、沙门氏菌都有较强的抑制作用。采用不同发酵时间的发酵液作为研究对象,测定发酵上清液的抑菌能力,发酵0-4 h上清液无抑菌能力,发酵至8 h抑菌能力逐步上升,发酵24-48 h发酵上清液抑菌能力趋于稳定,在48 h时抑菌能力最佳,抑菌直径为15.28mm。通过多元统计学分析乳酸菌发酵液差异标志物,发现主要差异物为有机酸(如乳酸、乙酸、丙酸等)和脂肪酸(如辛酸、癸酸等)。经过半制备液相分离发酵上清液得到的抑菌组分,主要有有机酸(如乳酸、乙酸、3-苯基乳酸、苯丙酸等)和脂肪酸(如癸酸、辛酸、壬酸等),另外还有少量的醛类和醇类物质。【结论】确定了植物乳杆菌DY6的抑菌物质主要为有机酸和脂肪酸,为其进一步防腐应用提供了理论基础。     

耐乙酸乳杆菌的筛选及其产乳酸特性

【背景】耐受乙酸的乳酸菌是传统谷物醋醋酸发酵过程中产生乳酸及其风味衍生物的重要功能微生物。【目的】从镇江香醋醋醅中分离鉴定具有耐乙酸特性的乳酸菌,并评价不同条件下该菌株的产乳酸能力。【方法】利用4%(体积比)乙酸含量的MRS培养基分离耐乙酸乳酸菌;对其进行16S rRNA基因鉴定、基因组测序、形态观察以及生理生化特性研究;考察不同乙酸浓度、葡萄糖浓度、发酵温度和时间对菌株产乳酸能力的影响。【结果】分离得到一株可耐受6%乙酸的乳杆菌Lactobacillus sp. JN500903;在厌氧静置、接种量5%、乙酸浓度5%、葡萄糖浓度40 g/L、发酵温度37°C、发酵时间10 d条件下,该菌株乳酸产量为16.1 g/L。【结论】乳杆菌JN500903能够耐受6%乙酸浓度,具有在酸性环境下合成乳酸的能力,有一定的应用潜力。     

Thin layer chromatographic determination of organic acids for rapid identification of bifidobacteria at genus level

This study presents a simple and fast method for the identification of bifidobacteria using a thin layer chromatographic (TLC) analysis of the short chain fatty acids in a culture broth. When the chromatogram was sprayed with the indicator solution (methyl red-bromophenol blue in 70% ethanol), lactic acid exhibited two red spots, and acetic acid, propionic acid, and butyric acid all produced blue spots. Succinic acid and citric acid produced yellow and dark yellow spot, respectively. In addition, these organic acids showed different R(f) values. The total time taken to analyze the organic acids in the 10 bacterial culture broths using the proposed method was approximately 50 min. The proposed TLC method was used to analyze the organic acids in culture broths of the following strains, five Bifidobacterium species. (Bifidobacterium longum, B. breve, B. infantis, B. bifidum, and B. adolescentis) and five other lactic acid bacteria strains (Lactobacillus casei, L. bulgaricus, L. acidophilus, Streptococcus thermophilus, and S. lactis). Both spots of lactic acid and acetic acid were detected on all the TLC plates from the five bifidobacterial culture broths. The five other lactic acid bacterial culture broths, however, only exhibited lactic acid spots. Accordingly, the proposed TLC method would appear to be a useful tool for rapid identification of Bifidobacterium spp. at the genus level.