南山牧场牛乳中具有抑菌活性乳酸菌的筛选及鉴定

该研究采用湖南南山高山牧场的鲜牛奶为原料,李斯特菌为指示菌,抑菌圈直径为评价指标,从中筛选抑菌特性较高的乳酸 菌,通过形态观察和分子生物技术对其进行鉴定。 同时,对菌株所产的抑菌物质的成分进行初步分析,并研究pH、温度及NaCl对其抑 菌效果的影响。 结果表明,从湖南南山高山牧场的鲜牛奶中共分离出28株乳酸菌,其中7株对李斯特菌具有抑菌活性,且菌株C15的 抑菌能力最强,抑菌圈直径为6.17 mm。 菌株C15被鉴定为乳酸乳球菌（Lactococcal lactis）,初步判定其所产的抑菌物质成分为蛋白类 或多肽类物质,该抑菌物质在酸性及中性条件下稳定,且具有较好的热稳定性和耐盐性,经80 ℃处理20 min后,抑菌活性保留54.35%; 经10%NaC（l 1 mol/L）处理2 h后,抑菌活性保留86.76%。     

红豆杉内生放线菌的分离及活性菌株的筛选与鉴定

通过组织匀浆法分离红豆杉(Taxus chinensis)根、茎、叶3个部位的内生放线菌,采用滤纸片法初步研究了内生放线菌发酵液的抑菌活性,并对抑菌活性明显的菌株进行了分子鉴定。结果表明,分离纯化得到内生放线菌55株,抑菌活性试验中共有21株表现出了不同程度的拮抗作用,占全部分离菌株的38.2%,其中4株具有明显的抑菌活性。经16SrDNA序列分析,初步鉴定3株为链霉菌属(Streptomyces sp.),1株为小单孢菌属(Micromonosporasp.)。     

贝莱斯芽孢杆菌抑尖孢镰刀菌脂肽类物质的鉴定

为探究贝莱斯芽孢杆菌（Bacillus velezensis）P9对于尖孢镰刀菌（Fusarium oxysporum）具有拮抗作用的主要活性成分,采用酸沉淀法和有机溶剂沉淀法提取菌贝莱斯芽孢杆菌P9发酵液中的脂肽类抗生素粗提物,研究粗提物对尖孢镰刀菌的抑菌活性。利用多功能制备液相仪对其组分进行分离,确定具有抑菌活性的组分,并利用高效液相色谱-四级杆飞行时间质谱联用（HPLC-Q-TOF-MS）仪对具有抑菌活性的组分进行鉴定。结果表明,脂肽类抗生素粗提物对尖孢镰刀菌有良好的抑菌作用,且酸沉淀法得到的粗提物抑菌活性较高,从中共分离到7种组分（编号为1#～7#）,其中4种组分具有抑菌作用,经鉴定,组分2#为C15SurfactinA或C16SurfactinB或C15SurfactinC,组分4#、6#、7#分别为C15BacillomycinD、C14BacillomycinD、C16BacillomycinD。     

华贵栉孔扇贝肠道产细菌素乳酸菌的分离筛选

采用双层牛津杯扩散法,以单核细胞增生李斯特氏菌、副溶血性弧菌、金黄色葡萄球菌、大肠埃希氏菌和枯草芽孢杆菌为指示菌,测定南海海域华贵栉孔扇贝肠道中分离到的乳酸菌的抑菌活性。对具有抑菌活性菌株的发酵液进行排酸、排过氧化氢实验,及胃蛋白酶、胰蛋白酶、木瓜蛋白酶敏感实验以筛选产细菌素菌株,并对细菌素进行温度、酸碱稳定性研究。结果显示:菌株ZY-F2的发酵液对食品中常见的腐败菌、致病菌有良好的抑制作用,蛋白酶酶解实验发现其抑菌物质具有蛋白质性质,初步确定为细菌素。该菌株发酵液经121℃处理15 min后相对抗菌活性为81.54%,且在pH2.0～6.0酸性范围内表现出抑菌活性。根据形态、生理生化鉴定及16S rRNA序列同源性分析,菌株ZY-F2被鉴定为食物魏斯氏菌(Weissella cibaria)。     

抗耐甲氧西林金黄色葡萄球菌菌株的筛选、鉴定及其活性物质初步研究

从海南红树林土壤分离筛选抗耐甲氧西林金黄色葡萄球菌（MRSA）菌株,通过形态学观察、生理生化试验及分子生物学进行菌种鉴定。采用不同培养基优选抑菌活性物质生成条件;以有机溶剂萃取抑菌活性物质,采用气相色谱-质谱（GC-MS）分析抑菌物质化学成分,采用微量稀释法测定最低抑菌浓度与最低杀菌浓度。结果表明,筛选得到一株抗MRSA菌株,编号为NEL-002,被鉴定为萎缩芽孢杆菌（Bacillus atrophaeus）。菌株NEL-002采用LB液体培养基得到发酵液经石油醚萃取粗提物抗MRSA菌活性最强,且在40～80 ℃、pH在5～10较稳定,对多种蛋白酶不敏感;最低抑菌质量浓度为62.5 μg/mL,最低杀菌质量浓度为500 μg/mL。GC-MS鉴定出抗菌活性物质为十甲基环戊硅氧烷、十二甲基环六硅氧烷及邻苯二甲酸二丁酯。     

产细菌素乳酸菌的筛选及鉴定

从传统发酵制品中分离筛选到50株乳酸菌,通过琼脂扩散法从中筛选出5株对指示菌具有明显抑菌作用的菌株,通过排除酸和过氧化氢的作用后,其中一株菌仍具有一定的抑菌活性.用胰蛋白酶对其发酵液进行处理后.活性下降很大,说明代谢产物中含有蛋白性质的抑菌物质,可能是细菌素.对此菌株进行鉴定,初步认为是戊糖乳杆菌,并对其生长能力和产酸能力进行了测定.     

蜂蜜中细菌的筛选、鉴定及其抑菌作用

从4种蜂蜜中分离得到8株芽孢杆菌,通过生理生化实验及形态学观察,结合16S rRNA基因序列分析对这些菌株进行了鉴定。初步研究了8株分离菌对大肠杆菌(Escherichia coli)、痢疾杆菌(Shigella dysenteriae S7NaCl)、粘质沙雷氏菌(Serratia marcescens SYBCT02)的抑菌作用。通过对这些分离菌的抑菌性质的研究,发现有些菌株是通过竞争作用抑制其他菌落的生长,还有些菌株是通过其代谢过程产生的抑菌物质来抑制其他菌落的生长。     

传统浆水中乳酸菌的筛选及抑菌性能分析

以甘肃传统发酵浆水为材料,采用平板划线法分离纯化其中的乳酸菌,并对纯化后的菌株进行形态学观察及生物学特性分析,筛选出其中具有抑菌活性的菌株,分别研究它们的抑菌能力以及抑菌物质的成分。结果表明:6株乳酸杆菌中,菌株SJ-1、SJ-2的发酵代谢产物对指示菌具有较高的抑菌活性。排除酸抑制作用和过氧化氢抑制作用后代谢产物仍具抑菌活性,且显示出良好的热稳定性。使用不同的蛋白酶对代谢产物进行处理后,其抑菌能力产生了程度不等的降低,表明抑菌物质具蛋白酶敏感性,因此判断抑菌物质中含有蛋白类抑菌成分。经鉴定发现菌株SJ-1为发酵乳杆菌(Lactobacillus fermentum),SJ-2为植物乳杆菌(Lactobacillus plantarum)。     

地衣腌菜中产广谱细菌素乳酸菌的筛选与鉴定

采用滤纸片双层平板法从自制酸奶、市售散装腌菜和袋装榨菜中分离出乳酸菌,再对分离的乳酸菌进行抑菌试验、酸和过氧化氢抑制作用的排除、蛋白酶的敏感检测和抑菌谱测定,最后对所得菌株形态和生理生化特性进行鉴定。结果:从样品中筛选到5株具有较高抑菌活性的乳酸菌菌株,在排除了酸和过氧化氢干扰后,发酵上清液仍有很强的抑菌作用;经蛋白酶处理后,发酵液的抑菌活性显著下降,说明该抑菌物具有蛋白质性质,是一类细菌素。这5株菌对绿脓假单胞杆菌、金黄色葡萄球菌、枯草芽孢杆菌和大肠埃希氏菌均有很强的抑菌活性,并选出抑菌活性最强的菌株E2-2,初步鉴定其为戊糖乳杆菌。     

罗布麻内生细菌的分离鉴定及抑菌活性

目的:研究罗布麻内生细菌及其抑菌活性,旨在深入开发新资源。方法:以红花罗布麻为研究对象,采用平板分离法从中分离内生细菌,利用牛津杯法对其发酵物进行抑菌活性研究,对有抑菌活性的菌株进行形态特征观察和16S rRNA序列同源性分析。结果:筛选出的D1菌株对大肠杆菌、金黄色葡萄球菌、藤黄微球菌和枯草芽孢杆菌四株供试指示菌均有明显的抑制作用,D3菌株对藤黄微球菌和枯草芽孢杆菌有一定的抑菌作用。结合菌株的形态学特征和16S rRNA序列同源性分析,将D1鉴定为假单胞菌,D3鉴定为菠萝泛菌。结论:罗布麻中蕴含着具有抑菌活性的内生细菌资源,可进一步的开发利用。     

Antibacterial and antioxidant properties of <Emphasis Type="Italic">Ramulus Cinnamomi</Emphasis> using supercritical CO<Subscript>2</Subscript> extraction

In this study, crude extracts of Ramulus Cinnamomi from supercritical carbon dioxide under various extraction conditions were examined for their antioxidant and antibacterial activity. The extractions were conducted in the range of 4,000–6,000 psi and 40–50 °C, and the solvent to feed ratio of the extraction was 30. The antibacterial activity was tested on the clinical drug-resistant strains, including 27 Acinetobacter baumannii, 20 Pseudomonas aeruginosa and 2 Staphylococcus aureus isolates by the disk diffusion method. The bioassay results indicated that Ramulus Cinnamomi showed obvious antimicrobial activity against the tested strains. This study also found that increasing the temperature and pressure would increase the yield of the supercritical fluid extraction (SFE), even though the best extraction conditions for antibacterial activity were found to be high pressure and low temperature. The minimum inhibitory concentration (MIC) was determined on the crude extract of Ramulus Cinnamomi, indicating that the crude extracts from supercritical extraction showed better antibacterial activity than those obtained by ethanol extraction. Based on the spectrophotometer and bioassay determination, the antimicrobial constituent was identified to be cinnamaldehyde. Time-kill kinetics and scanning electron microscopy (SEM) were employed to monitor the survival characteristics and the changes in morphologies, respectively, of the test microorganisms in the presence of herbal extracts. Moreover, antioxidant activity was evaluated by scavenging of the free radical DPPH. Extracts of Ramulus Cinnamomi provided 50% inhibition at 2 mg/ml concentration. This study will provide valuable information for extraction of the natural bioactive component, cinnamaldehyde, from Ramulus Cinnamomi by supercritical extraction.     

黑曲霉孢子粉粗提物对青枯雷尔式菌的抑菌机制初探

以黑曲霉（Aspergillus niger）孢子粉作为原料,对其粗提物进行分离,以青枯雷尔式菌（Rastonia solanacearum）为指示菌,对抑菌活性进行监测,并利用气相色谱-质谱联用（GC-MS）技术分析其活性成分。通过测定电导率、核酸蛋白泄露、总蛋白以及活性氧（ROS）等指标的变化分析黑曲霉抑菌机制。结果显示,黑曲霉粗提物洗脱液S5对青枯雷尔式菌具有良好的抑菌效果,抑菌率可达90.87%;GC-MS检测结果显示,其主要成分为油酸（26.52%）、亚油酸甲酯（14.46%）、油酸甲酯（11.10%）、棕榈酸（10.50%）、油酸乙酯（7.36%）、亚油酸乙酯（6.88%）等;粗提物主要通过影响细胞膜完整性、损伤细胞结构、抑制细胞蛋白质合成及能量代谢等方面达到抑制菌株Q11-2的效果。     

合浦珍珠提取物对金黄色葡萄球菌的体外抑菌活性及稳定性研究

目的:研究合浦珍珠提取物对金黄色葡萄球菌的体外抑菌活性及稳定性。方法:采用牛津杯法及96孔法测定合浦珍珠提取物对金黄色葡萄球菌的抑菌活性及最小抑菌浓度,并探讨温度、反复冻融、酸碱、紫外线、金属离子、胰蛋白酶和胃蛋白酶对合浦珍珠提取物抑菌活性的影响。结果:合浦珍珠提取物对金黄色葡萄球菌具有显著的抑菌活性,最小抑菌浓度为0.94 mg·mL-1,且抑菌活性呈一定的剂量依赖效应。以8×MIC合浦珍珠提取物进行抑菌稳定性实验,以金黄色葡萄球菌的OD₆₀₀值为指标进行探讨,结果显示,合浦珍珠提取物在pH3.0~9.0抑菌活性保持稳定,但在pH11.0相比pH7.0抑菌活性,保留约70%;在-80~80 ℃抑菌活性保持稳定,但在100 ℃相比4 ℃处理30 min抑菌活性保留约68%;在-20 ℃反复冻融及20 W紫外灯照射条件下,提取物抑菌活性保持良好的稳定性;在25~200 mmol·L-1的NaCl、KCl、CaCl₂条件下,提取物抑菌活性稳定,但在25~200 mmol·L-1MgCl₂条件下,抑菌活性逐渐降低,在200 mmol·L-1 MgCl₂时,抑菌活性相比未添加金属离子组保留约40%;提取物对胰蛋白酶处理保持良好的抑菌稳定性,但在胃蛋白酶处理下,抑菌活性消失,说明对胃蛋白酶敏感。结论:合浦珍珠提取物对金黄色葡萄球菌具有稳定的体外抑制活性,为其今后在食品工业上的应用和开发提供一定的理论基础。     

苦荞麸皮黄酮提取物及有效成分的抑菌活性

研究苦荞麸皮总黄酮粗品及精品对病原菌的抑制作用,并初步分析其黄酮类成分,评价不同黄酮单体的抑菌活性。通过平板打孔法及二倍稀释法检测苦荞麸皮总黄酮粗品及精品及其4种黄酮单体的抗菌活性,使用高效液相色谱法定性定量检测苦荞麸皮总黄酮粗品及精品的黄酮成分。结果表明,苦荞麸皮黄酮精品对金黄色葡萄球菌、藤黄微球菌、肠球菌、大肠杆菌、志贺氏杆菌、沙门氏菌的抑制效果优于粗品。苦荞麸皮黄酮主要黄酮成分为芦丁、山奈酚-3-O-芸香糖苷、槲皮素和山奈酚,精品的总黄酮含量是粗品总黄酮含量的2.84倍。4种黄酮单体的抑菌能力具有一定的构效关系,其中槲皮素对所有供试菌均表现出较好的抑菌活性。     

火炭母提取物对金黄色葡萄球菌的抑菌活性及稳定性研究

目的：研究火炭母提取物对金黄色葡萄球菌的抑菌活性及稳定性。方法：用牛津杯法和微量肉汤稀释法来测定火炭母提取物的体外抑菌活性,并以金黄色葡萄球菌的OD_{600 nm}值为指标,进一步探讨温度、pH、紫外线及Na⁺、K⁺、Ca²⁺等金属离子对其抑菌稳定性的影响。结果：火炭母提取物对金黄色葡萄球菌具有一定抑菌活性,最小抑菌浓度为8 mg/mL;火炭母提取物在热处理、不同pH、紫外线照射等情况下抑菌活性较为稳定;当添加不同金属离子时其抑菌活性具有明显差异,0.1 mol/L Na⁺、K⁺处理时其抑菌活性有所下降,而在0.1 mol/L Ca²⁺处理时其抑菌活性丧失。结论：火炭母提取物对金黄色葡萄球菌具有稳定的体外抑菌活性。     

Chemical profiling of Punica granatum peels from Jordan using LC–MS/MS and study on their biological activities

A paucity of local information concerning the chemical profiles and biological activities of extracts obtained from less-studied P. granatum peel of Jordanian origin was considered in this study for the first time. Fractionation of the crude ethanol extracts was performed because of their higher phenolic and flavonoid contents compared to the water and acetone extracts. The chemical compositions of the respective samples, that is, extracts/fractions were identified by LC–MS/MS, and the elemental content of the raw materials was also analysed using atomic absorption spectrometry. Antioxidant activities of extracts and fractions were evaluated against DPPH and ABTS radical scavenging assays, and the antibacterial activities were investigated by disc diffusion method and MIC (Minimum Inhibitory Concentration). The LC–MS/MS results correlated strongly with the total phenolic and flavonoid contents, where ethanol displayed higher efficacy for extracting bioactive ingredients. Overall, a total of 19 phenolics were detected in the ethanolic peel extract of P. granatum. All of the analysed extracts showed strong antioxidant activities. Fractionation of ethanolic extracts resulted in fractions with almost similar chemical LC–MS/MS profiles, but the relative proportion of constituents was different. All fractions showed lower intensities of antioxidant capacities than crude extracts, highlighting the superiority of some components, either individually or combined, as well as their relative proportion on the biological activity of each fraction. The results of the present study emphasise the need to further explore the effect of putative interactions among plant bioactive ingredients and focus on possible interactions with drugs along with foods.     

Dunaliella salina microalga pressurized liquid extracts as potential antimicrobials

In the present work, the antimicrobial activity of different pressurized liquid extracts obtained from Dunaliella salina microalga was tested against several microorganisms of importance for the food industry (Escherichia coli, Staphylococcus aureus, Candida albicans, and Aspergillus niger). Different solvents (hexane, petroleum ether, hexane, and water) and extraction conditions (40, 100, and 160 degrees C) were tested. Results showed that the best antimicrobial activity was obtained for each solvent at the highest extraction temperature (160 degrees C). Likewise, the extraction yield followed the same trend, i.e., increasing with extraction temperature and was at a maximum when ethanol was used as an extraction solvent. Water extracts had the lowest extraction yields. In general, the best results in terms of antimicrobial activity were obtained using petroleum ether and hexane, although ethanolic extracts also showed good antimicrobial activity. Because the main antimicrobial activity of the extracts was against bacteria, the extracts can be considered to be specifically antibacterial. The extracts were analyzed by gas chromatography-mass spectrometry in order to identify the compounds responsible for activity. Fifteen different volatile compounds as well as several fatty acids (mainly palmitic, alpha-linolenic, and oleic acids) that could have been responsible for the antimicrobial activity were identified in the extracts. beta-Cyclocitral, alpha- and beta-ionone, neophytadiene, and phytol were identified among other volatile compounds; all of these compounds have previously been described as antimicrobial agents.     

抗耐甲氧西林金黄色葡萄球菌菌株的筛选与鉴定

通过对246株芽孢杆菌进行筛选,以期获得对耐甲氧西林金黄色葡萄球菌（MRSA）具抑制作用的菌株,抑菌结果表明,对耐甲氧西林金黄色葡萄球菌有抑制作用的菌株有5株,其中菌株1012-4的抑菌活性最强;为确定菌株种属,对优选菌株进行形态特征观察、生理生化特征鉴定以及API50 CHB鉴定,初步确定1012-4为枯草芽孢杆菌;为进一步确证菌株类型,实验采用了16S rDNA基因测序,序列结果表明,菌株1012-4为芽孢杆菌属枯草芽孢杆菌种;经反复确证,确定菌株1012-4为优势菌株,抗菌活性显著,为新型抗MRSA药物的研发提供了菌株来源。