云南省4种蕨类植物提取液的抑菌活性

为了合理开发利用蕨类植物资源,本研究利用圆纸片法分析了云南省4种蕨类植物单芽狗脊蕨（Woodwardia unigemmata）、蜈蚣蕨（Pteris vittata）、鸡足山耳蕨（Polystichum jizhushanense）和灰绿耳蕨（Polystic-hum eximium）的提取液对金黄色葡萄球菌（Staphylococus aureus）、大肠杆菌（Escherichia coli）和枯草芽孢杆菌（Bacillus subtilis）的抑菌活性。抑菌试验结果表明,当4种蕨类植物提取液浓度分别为10%、8%、6%和4%时,对照组无抑菌圈出现,单芽狗脊蕨提取液对大肠杆菌和金黄色葡萄球菌无抑菌活性,灰绿耳蕨提取液对金黄色葡萄球菌无抑菌活性,其它的蕨类植物提取液对供试微生物都表现出不同程度的抑菌活性;并且这4种蕨类植物提取液至少对一种供试细菌具有抑菌活性,抑菌活性范围为8～14mm,平均抑菌圈直径为11.2mm,4种提取液对枯草芽孢杆菌抗菌活性效果最明显。由此,我们推测本研究中的4种蕨类植物均有抗菌物质存在,并将有望成为筛选抗菌新型药物的种质资源。     

蒲公英不同部位抑菌作用及代谢物差异的UPLC-MS/MS分析

为了研究蒲公英不同部位抑菌作用和代谢物成分的差异,本物采用打孔法和超高效液相色谱串联质谱(UPLC-MS/MS)方法,研究了蒲公英根、茎叶和花的水提取物和醇提取物对5种致病菌的抑菌作用,并对不同部位的代谢物成分进行了定性和定量分析。结果表明,蒲公英不同部位初提取物对变形杆菌、嗜根考克氏菌和枯草芽孢杆菌均有抑菌作用,而对大肠杆菌和铜绿假单胞菌均无抑菌作用;醇提法的抑菌效果优于水提法,花提取物的抑菌活性最强,根最弱。蒲公英的不同部位共鉴定出449种代谢物,根、茎叶和花中相对含量最高的物质分别为反油酸(脂类)、γ-亚麻酸(脂类)和木犀草素(黄酮类);且根、茎叶和花中相对含量排名前20的物质中黄酮类分别有0种、3种和8种,表明黄酮类物质可能是蒲公英中的主要抑菌活性物质。正交偏最小二乘法-判别分析(OPLS-DA)结果显示,木犀草素-7-O-葡萄糖苷在花中上调幅度最大,橙皮素C-丙二酰己糖苷为根中特有物质。本研究结果为利用蒲公英不同部位研制和开发新型药物提供了理论依据。     

利用环介导等温扩增技术快速检测水产品中的副溶血弧菌

副溶血弧菌(Vibrio parahaemolyticus)是一种重要的食源性致病菌。首次将环介导等温扩增技术（Loop-Mediated Isothermal Amplification, LAMP）应用于副溶血弧菌的快速检测。针对副溶血弧菌不耐热溶血毒素基因（tlh）设计四条特异性引物,建立了副溶血弧菌LAMP检测方法,1 h即可完成。对12种细菌共28株菌进行LAMP扩增,仅副溶血弧菌得到阳性结果,证明引物具有很高的特异性;副溶血弧菌基因组DNA和纯培养物的检测灵敏度分别为90 fg/LAMP反应体系和24 cfu/mL,对模拟食品样品进行直接检测,检测限为89 cfu/g;对40份水产样品进行检测,8份副溶血弧菌LAMP阳性,其中6份传统培养阳性。本研究表明,该方法检测副溶血弧菌特异性强、灵敏度高,并且操作简便、快捷、检测成本低,有望发展成为快速检测副溶血弧菌的有效手段。     

苹果渣中不同极性多酚的分离及体外抗氧化活性研究

为了优化苹果多酚提取工艺,进而提高苹果多酚提取物中强效多酚的含量,该文采用液-液萃取方法按极性大小对果渣多酚进行分离,进而采用1,1-二苯基-2-苦苯肼自由基法和铁离子还原/抗氧化力法对不同极性多酚体外抗氧化活性进行评价.结果表明:液-液萃取方法,可以实现不同极性多酚的有效分离,不同极性苹果多酚,其体外抗氧化活性有显著差异,水层多酚活性最强,其次为正丁醇层,乙酸乙酯层相对较弱,即多酚的抗氧化性与其极性呈正相关.因此,工业化提取苹果渣中多酚时,增大提取溶剂的极性有利于强效多酚的获取.     

壳聚糖酶基因的克隆与序列分析

本研究采用一对引物从巨大芽孢杆菌BS-0409中成功地克隆了壳聚糖酶基因（Csn） 全基因序列,大小为516bp,将其在NCBI网站上用BLAST程序进行在线检索分析,结果与多种芽孢杆菌同源性均为96%。同时,利用DNAMAN软件比较巨大芽孢杆菌BS-0409与其它13种不同细菌Csn编码基因的氨基酸序列,结果显示不同细菌Csn氨基酸序列之间有比较高的同源性,且与多种芽孢杆菌具有较高的同源性。     

复合益生菌发酵液的功能特性及对对虾诱食效果

为探求复合菌发酵液的多种功能,该试验选择具有脱氮、产酶、抑菌等优良性能的酵母菌、乳酸菌及芽孢杆菌各1株,利用已优化的HJ培养基进行共培养,实时监测发酵过程,分时段取样,对复合菌发酵液的脱氮、产酶、抑菌、培藻及诱食等功能进行研究。结果表明,酿酒酵母菌NJ-02、屎肠球菌SC-01及枯草芽孢杆菌M7-1能够在HJ培养基中进行共发酵,连续培养24 h后3株微生物活菌数量分别达到3.88×10~8、2.41×10~(10)、5.38×10~9 CFU/mL。复合菌发酵液的脱氮、培藻性能同复合菌中枯草芽孢杆菌M7-1的活菌数相关较大,发酵至16h其降解亚硝态氮和培藻性能最为理想,亚硝态氮降解率为89%,并使小球藻叶绿素a质量分数提升49.6%。复合菌发酵液具有同枯草芽孢杆菌M7-1相当的产酶(蛋白酶、淀粉酶、纤维素酶)活性及同屎肠球菌SC-01相当的抑菌(副溶血弧菌)活性。复合菌发酵液饲喂对虾,诱食效果明显好于对照组(P0.05),同化学诱食剂二甲基-β-丙酸噻亭(dimethyl-beta-propiothetin hydrochloride, DMPT)差异不显著,其肠道中乳酸菌、酵母菌数目显著高于对照组及化学诱食剂组(P 0.05)。该研究所制备的脱氮、培藻、抑菌及诱食功能复合菌发酵液,为可持续生态水产养殖提供了新的微生物资源和技术方法。     

竹黄无性型菌株液体发酵菌丝提取物的生物活性研究

测定竹黄无性型菌株ZHLS-03发酵菌丝体中竹红菌甲素的含量,并对其菌丝提取物的清除自由基活性和抑菌活性进行了研究。按50μg/ml的浓度测定了菌丝体石油醚、乙酸乙酯和甲醇等3种提取物对水稻纹枯病菌、稻瘟病菌、棉花枯萎病菌3种植物病原菌以及白色假丝酵母、金黄色葡萄球菌2种病原细菌的抑制能力;同时测定菌丝体的石油醚、乙酸乙酯和饱和正丁醇等3种提取物对DPPH自由基的清除率。结果表明：菌株ZHLS-03液体培养至6d,菌丝体中竹红菌甲素含量最高,可达到28029mg/g;菌丝体甲醇和正丁醇提取物分别具有较强的抑菌活性和清除自由基活性,甲醇相对棉花枯萎病菌的抑制率为7478%,抗白色假丝酵母的抑菌圈直径达到2069mm,且反应15min后正丁醇提取物对DPPH自由基的清除率可达9324%。     

入侵植物假臭草和胜红蓟提取物的抑菌活性研究

假臭草和胜红蓟是我国南方常见的入侵植物,为了防治和利用其开发植物源抑菌剂,本试验采用牛津杯法检测了不同溶剂(70%乙醇、95%乙醇、石油醚和乙酸乙酯)提取物对28种病原微生物的抑菌活性,并评估了不同器官部位提取物的抑菌活性。结果表明,不同溶剂提取物抑菌活性有差异,假臭草70%乙醇提取物对金黄色葡萄球菌(Staphylococcus aureus)抑制活性最强,抑菌圈直径为26.28±0.75mm;胜红蓟95%乙醇提取物对腐生葡萄球菌(Staphylococcus epidermidi)抑制活性最好,抑菌圈直径为17.31±0.46 mm。此外,假臭草和胜红蓟70%乙醇不同部位提取物对金黄色葡萄球菌(Staphylococcus aureus)的抑菌作用表现为叶部提取物(22.31±0.34 mm和19.52±0.28 mm)茎部提取物(22.25±0.65 mm和12.75±0.45 mm)根部提取物(11.78±0.69 mm和9.05±0.53 mm)。假臭草提取物对12种细菌和1种真菌黑曲霉菌(Aspergillus niger)具有抑制效果;胜红蓟提取物对9种细菌具有抑制效果。本研究结果为进一步研发具有潜力的植物源抑菌剂奠定了理论基础。     

硫酸盐还原细菌（SPBs）的筛选培养及在两相厌氧工艺中试验研究

通过硫酸盐还原细菌（ＳＰＢｓ）的筛选、培养条件优化及在两相厌氧工艺中产酸相应用硫酸盐还原细菌（ＳＰＢｓ）试验,选出了一株耐富含硫酸盐高浓度酵母生产废水的脱硫弧菌,并强化两相厌氧的工艺中产酸相硫酸盐还原细菌（ＳＰＢｓ）的活性。     

紫苏挥发油化学成分GC/MS分析及抑菌评价研究

采用水蒸汽蒸馏法分别提取紫苏叶和花中挥发油,运用毛细管气相色谱-质谱联用法结合计算机检索对其挥发油进行化学成分分析,采用面积归一法测定了挥发油中各种成分的相对质量分数,利用抑菌贴片法、MIC法和光电浊度法分别对枯草芽孢杆菌和大肠杆菌进行了抑菌性研究。结果显示,从紫苏叶和花水蒸汽蒸馏所得挥发油中分别检测出12种和10种化合物,其中叶中主要成分为紫苏醛（75.88%）、柠檬烯（3.82%）、石竹烯（9.87%）和法呢烯（3.21%）;花中主要成分为紫苏醛（49.23%）、石竹烯（18.53%）、法呢烯（8.99%）、芳樟醇（11.3%）和姜黄二酮（4.55%）。紫苏精油对两种细菌均有很强的抑制作用,但对枯草芽孢杆菌的抑制能力要明显强于对大肠杆菌的抑制能力。     

商陆根部多糖的分离、纯化及其体外抗氧化活性

本研究从商陆根部提取分离出商陆多糖,并对该多糖含量进行测定及体外抗氧化活性研究。商陆根部经石油醚回流脱脂、热水抽提、脱色、乙醇沉淀等处理可从中提取出粗多糖。多糖经三氯乙酸脱蛋白纯化,再分别以考马斯亮蓝法和蒽酮-硫酸法测定其蛋白质含量和糖含量,并通过采用清除羟自由基和超氧自由基能力测定法、β-胡萝卜素-亚油酸法,以抗坏血酸（Vc）为对照,来分析粗多糖和纯多糖的体外抗氧化能力。采用水提法提取的粗多糖,含有10.69%的蛋白质和60.91%的多糖;纯化后的多糖糖含量达到91.04%。商陆粗多糖、纯多糖及Vc对羟自由基（·OH）清除率的IC50分别是1.26mg/mL、4.78mg/mL和0.224mg/mL;对超氧自由基（O2-·）清除率的IC50分别为1.91mg/mL、2.28mg/mL和0.123mg/mL;对β-胡萝卜素-亚油酸体系的抑制作用的IC50分别为0.471mg/mL、0.692mg/mL和0.379mg/mL。实验结果表明商陆多糖是一种较好的抗氧化剂,可作为潜在天然抗氧化剂应用于食品和医药工业中。     

In vitro cytotoxicity of nonpolar constituents from different parts of kava plant (Piper methysticum)

Kava (Piper methysticum), a perennial shrub native to the South Pacific islands, has been used to relieve anxiety. Recently, several cases of severe hepatotoxicity have been reported from the consumption of dietary supplements containing kava. It is unclear whether the kava constituents, kavalactones, are responsible for the associated hepatotoxicity. To investigate the key components responsible for the liver toxicity, bioassay-guided fractionation was carried out in this study. Kava roots, leaves, and stem peelings were extracted with methanol, and the resulting residues were subjected to partition with a different polarity of solvents (hexane, ethyl acetate, n-butanol, and water) for evaluation of their cytotoxicity on HepG2 cells based on the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and lactate dehydrogenase and aspartate aminotransferase enzyme leakage assays. Organic solvent fractions displayed a much stronger cytotoxicity than water fractions for all parts of kava. The hexane fraction of the root exhibited stronger cytotoxic effects than fractions of root extracted with other solvents or extracts from the other parts of kava. Further investigations using bioassay-directed isolation and analysis of the hexane fraction indicated that the compound responsible for the cytotoxicity was flavokavain B. The identity of the compound was confirmed by (1)H and (13) C NMR and MS techniques.     

铁锰营养失衡对商陆根系分泌物的影响

以商陆为实验材料,用水培方法设置了8个实验处理:Fe0Mn0(对照),Fe0Mn-,Fe0Mn ,Fe-Mn0,Fe-Mn-,Fe-Mn ,Fe Mn0,Fe Mn-,Fe Mn 。处理30 d时,对根系分泌物中可溶性糖、总氨基酸、pH值以及根系质膜透性进行测定。结果表明,Fe-Mn-的可溶性糖含量稍高于对照,其余各处理组均低于对照,Fe0Mn ,Fe Mn 达到显著水平;氨基酸含量在铁锰失衡条件下普遍有所增加,其中Fe-Mn0分泌的氨基酸含量最大,与对照差异显著;除Fe0Mn 外,各处理组的pH值与对照组相比都显著增加,其中以铁毒处理组(Fe Mn0,Fe Mn ,Fe Mn-)的增加率最高;铁毒处理组(Fe Mn0,Fe Mn ,Fe Mn-)使商陆根系质膜透性增加,对商陆根系造成一定的伤害。总体而言,铁锰的缺乏或过量都会引起商陆根系分泌物含量的变化,其中以铁胁迫和锰毒对其影响最大;另外,相对单铁或单锰处理而言,铁锰之间的交互作用对商陆根系分泌物的变化也有一定的影响。     

Bioactive aromatic compounds from leaves and stems of Vanilla fragrans.

Alcoholic extracts of leaves and stems of Vanilla fragrans were fractionated with ethyl acetate and aqueous butanol. All three fractions of ethyl acetate, butanol, and water were screened for toxic bioactivity against mosquito larvae. The results of these experiments showed that the fractions from the ethyl acetate and butanol phases were both active in the bioassay. Bioactivity of the ethyl acetate fraction was found to be much greater than that from the butanol fraction in mosquito larvae toxicity. The water phase appeared to contain no substances that impaired mosquito larval growth. Repeated column chromatography of the ethyl acetate fraction on silica gel led to the isolation of 4-ethoxymethylphenol (1), 4-butoxymethylphenol (2), vanillin (3), 4-hydroxy-2-methoxycinnamaldehyde (4), and 3,4-dihydroxyphenylacetic acid (5). Compounds 4 and 5 were isolated from Vanilla species for the first time and 2 has not been reported to have been found in a natural form. 4-Ethoxymethylphenol (1) was the predominant compound, but 4-butoxymethylphenol (2) showed the strongest toxicity to mosquito larvae. The structures of the compounds were determined on the basis of their mass spectra and (1)H or (13)C NMR data.     

Essential oil constituents and in vitro antimicrobial activity of Decalepis hamiltonii roots against foodborne pathogens

Hydrodistillation of Decalepis hamiltonii roots yielded an essential oil (0.33% v/w) that contained 2-hydroxy-4-methoxybenzaldehyde (37.45%), 2-hydroxybenzaldehyde (31.01%), 4-O-methylresorcylaldehyde (9.12%), benzyl alcohol (3.16%), and alpha-atlantone (2.06%) as major constituents, with aromatic aldehydes constituting the main fraction of this root's essential oil. The oil was tested for its antimicrobial activity against foodborne pathogens responsible for food spoilage and human pathologies using standard antimicrobial assays. It exhibited strong antimicrobial activity against Bacillus cereus, Bacillus megaterium, Candida albicans, Escherichia coli, Micrococcus luteus, Micrococcus roseus, and Staphylococcus aureus at a concentration range of 1:0 with inhibitory activities of 27, 23, 16, 19, 22, 19, and 23 mm, respectively, which are comparable to those of the standards. The roots of D. hamiltonii, therefore, may be considered as an inexpensive source of an essential oil rich in antimicrobial compounds against foodborne pathogens.     

锰尾渣改良对4种植物吸收锰的影响

采用盆栽法研究锰尾渣改良对商陆（Phytolacca acinosa）、美洲商陆（Phytolacca americana）、繁穗苋（Amaranthus paniculatus）和酸模叶蓼（Polygonum lapathif olium）4种植物吸收锰的影响。将锰尾渣与土壤混合进行基质改良,有利于植物生长。基质含80%锰尾渣时,商陆、美洲商陆和繁穗苋3种植物生长正常,吸收锰多。商陆和美洲商陆2种植物对锰尾渣耐受性强,生物量大,植物提取锰多;繁穗苋对锰尾渣耐受性较强。在锰尾渣污染区进行植物修复时,商陆和美洲商陆可作为首选植物,繁穗苋可作为辅助共生植物,采用锰尾渣含量为80%的基质改良方式最佳。     

Size Characterization by Laser Granulometry of Colloids Extracted from Compost at Different Temperatures

The objective of this work was to characterize colloids extracted from composts and their potential retention in soils. Compost made of sludge and green wastes was sampled (i) during the fermentation phase and (ii) after maturation. The same kind of compost was used in a long-term field experiment at Feucherolles (France), near Paris where amended and nonamended soils were sampled. The colloidal fraction was extracted from composts in water at room temperature (20°C) and compared to the colloidal fraction extracted from the soil. Composts were also extracted by pressurized hot liquid water at 50, 125 and 175°C. The total organic carbon of the extracts was measured and the particle size distribution (PSD) of colloidal extracts was analyzed by laser granulometry. The diameters of the colloids extracted from the soil ranged between 0.040 and 0.300 μm, independently of the temperature. For composts, it varied from 0.040 to 3.200 μm when extraction was done at 20°C, while at higher temperatures, much more organic matter was extracted, and colloid diameters ranged from 0.040 μm to 0.200 μm. The water-soluble C decreased and the size of colloids recovered in water at temperatures below 50°C increased when compost maturity increased. The adsorption on soils of colloidal particles extracted from composts was characterized. The largest adsorption (up to 30% of the initial soluble C) occurred with the extracts recovered at high temperature, in relation to the more hydrophobic properties of the colloids extracted with hot water maintained in subcritical conditions. After adsorption, the particle size distribution in the colloidal fraction extracted at 20°C moved towards finer fractions; by contrast, the colloidal fraction extracted at 175°C moved towards coarser fractions. The coarsest colloids coming from the soil disappeared during the adsorption experiment, probably because of the coprecipitation with the finest colloids coming from compost.     

Fractional extraction of compounds from grape seeds by supercritical fluid extraction and analysis for antimicrobial and agrochemical activities

White grape seeds were subjected to sequential supercritical fluid extraction. By increasing the polarity of the supercritical fluid using methanol as a modifier of CO(2), it was possible to fractionate the extracted compounds. Two fractions were obtained; the first, which was obtained with pure CO(2), contained mainly fatty acids, aliphatic aldehydes, and sterols. The second fraction, obtained with methanol-modified CO(2), had phenolic compounds, mainly catechin, epicatechin, and gallic acid. The fractions were bioassayed. Antimicrobial activities were checked on human pathogens, and a high degree of activity was obtained with the lipophilic fraction. Agrochemical activities on phytopathogenic fungi and activities on the etiolated wheat coleoptile bioassay were also checked. The more polar fraction was active in the latter bioassay.     

Phlorotannins from brown algae (Fucus vesiculosus) inhibited the formation of advanced glycation endproducts by scavenging reactive carbonyls

Accumulation of advanced glycation end products (AGEs) in vivo is associated with aging, diabetes, Alzheimer's disease, renal failure, etc. The objective of this study was to investigate the inhibitory effects of brown algae Fucus vesiculosus phlorotannins on the formation of AGEs. F. vesiculosus phlorotannins were extracted using 70% acetone. The resultant extract was fractionated into dichloromethane, ethyl acetate, butanol, and water fractions. The ethyl acetate fraction was further fractionated into four subfractions (Ethyl-F1 to -F4) using a Sephadex LH-20 column. F. vesiculosus acetone extract or fractions significantly inhibited the formation of AGEs mediated by glucose and methylglyoxal in a concentration-dependent manner. The concentrations of F. vesiculosus extracts required to inhibit 50% of albumin glycation (EC(50)) in the bovine serum albumin (BSA)-methylglyoxal assay were lower than those of aminoguanidine (a drug candidate for diabetic complication), except for F. vesiculosus acetone extract and dichloromethane fraction. In the BSA-glucose assay, F. vesiculosus extracts inhibited BSA glycation more than or as effectively as aminoguanidine, except for Ethyl-F3 and -F4. The ethyl acetate fraction and its four subfractions scavenged more than 50% of methylglyoxal in two hours. The hypothesis whether F. vesiculosus phlorotannins scavenged reactive carbonyls by forming adducts was tested. Phloroglucinol, the constituent unit of phlorotannins, reacted with glyoxal and methylglyoxal. Five phloroglucinol-carbonyl adducts were detected and tentatively identified using HPLC-ESI-MS(n).