云南3个盐矿真菌活性菌株的筛选

采用滤纸片法和胞外酶定性筛选培养基对云南3个盐矿(乔后盐矿、一平浪盐矿和昆明盐矿)真菌进行抗菌活性筛选和产胞外酶筛选,并通过显微形态特征的观察对活性菌株进行鉴定,同时对活性较好的菌株进行NaCl的耐受实验。结果筛选到有抗菌活性的菌株51株,其中有26株对白假丝酵母(Candida albicans)有抗菌活性;产胞外酶活性的菌株有67株,产纤维素酶、蛋白酶和淀粉酶的菌株各有58株、41株和26株。活性菌株的优势属均为青霉属Penicillium。活性菌株对NaCl有不同程度的耐受性。     

傣药植物内生真菌的抑菌活性评价及发酵条件探索

从云南傣药植物中分离到180株内生真菌,选用9种培养基进行发酵。利用茄腐镰刀菌、尖孢镰刀菌、立枯丝核菌、玉米小斑病、稻梨孢菌等5种植物病原菌作为指示菌株,结合TLC检测对其发酵粗提物进行活性评价和化学多样性分析,以期寻找到具有开发潜力的活性菌株。研究结果表明活性菌株为36株,其中有7株菌活性好且产物多样性丰富。     

灯台树内生放线菌多样性及抗菌活性评价

从云南西双版纳采集7份灯台树样品,经过表面消毒,用4种分离培养基分离得到105株内生放线菌。经16S rRNA基因序列分析鉴定,分布于7个科9个属。利用5种植物病原真菌指示菌对所有菌株的发酵液进行抗菌活性检测。结果显示,有12.4%、14.3%、11.4%、12.4%、8.6%的菌株分别对镰刀霉、疫霉、赤星霉、苹果炭疽、白色念珠菌有抗性。对3株具有广谱抗菌活性的菌株进行再次发酵和抗菌活性复筛,结果显示这3株的抗菌活性稳定,并可能含有生物碱类化合物。     

广西北部湾放线菌的分离筛选及活性产物的鉴定

为从广西北部湾的泥样和植物中分离海洋放线菌,筛选具有抑菌活性的菌株,分离活性化合物。研究采用普通稀释法分离菌株,对发酵产物进行抑菌活性测试,利用活性追踪分离活性化合物,并通过波谱方法确定化合物结构。结果表明从6个海泥样品和3个植物样品中共分离73株放线菌,筛选得到具有抑香蕉枯萎病和金黄色葡萄球菌活性的菌株7株,并从其中的1株链霉菌 Streptomyces sp.MDCW-126的次级代谢产物中分离鉴定了星形孢菌素。从广西北部湾分离的药用活性菌株资源具有开发和深入研究价值。     

茵陈内生细菌的分离鉴定与抑菌活性成分分析

为从中药茵陈中分离内生细菌,筛选具有抑菌活性的菌株,分析活性成分。研究采用7种不同的培养基分离菌株,对分离到的菌株进行形态学和分子生物学鉴定,并对发酵产物进行抑菌活性测定,并利用分光光度法和高效液相色谱法分析抑菌活性较高的菌株发酵液活性成分。结果表明从茵陈新鲜组织中共分离到52株内生细菌,分属于9个种,芽孢杆菌为优势菌群。筛选得到具有抑菌活性的菌株17株,其中NA-3、BPA-24和KMB-32 3株菌活性较强,NA-3和KMB-32的发酵液中齐墩果酸含量较高。本研究通过分离纯化茵陈内生细菌,明确了茵陈内生细菌的菌群类别,获得了产齐墩果酸的两个菌株,为微生物发酵生产活性物质提供了素材。     

海洋细菌抗菌和细胞毒活性的初步研究

从不同海域的生物、海水和海泥中分离海洋细菌,利用琼脂扩散法和MTT法对细菌培养液的乙酸乙酯提取物进行了抗菌和细胞毒活性筛选。比较了活性菌株与来源的相关性．结果表明,在分离的341株海洋细菌中。42株细菌的代谢产物具有抗菌活性,7株具有细胞毒活性,其中来源于海洋无脊椎动物和海藻的活性菌株比例(22％和11％)大于来源于海水和海泥的细菌(7％和5％)．细菌分类鉴定结果显示,具有活性的细菌大部分属于假单胞菌属、发光杆菌属、梭状芽孢杆菌属、交替单胞菌属和黄杆菌属．     

南方红豆杉内生及根际放线菌多样性及其生物活性

【目的】研究药用植物南方红豆杉内生及根际土壤放线菌的多样性及其抑菌、抗肿瘤等重要生物活性并获得一些具有强抑制植物病原真菌以及抗肿瘤等重要生物活性的菌株。【方法】选择7种培养基从南方红豆杉及其根际土壤中分离放线菌,对链霉菌进行形态学分类,去重复后对其进行抑制植物病原真菌以及抗肿瘤活性的筛选并对高活性菌株进行初步鉴定。对部分菌株进行16S rRNA基因测序分析研究其多样性。【结果】研究共分离得到277株放线菌,经去重复后剩余111株放线菌,可归类到6个亚目、7个科、8个属。其中链霉菌可分为10个类群。生物活性研究结果显示:30.9%的菌株具有抑制植物病原真菌活性,其中6株放线菌对多种植物病原真菌显示了强的抑菌活性。分别有44.1%和33.3%的菌株对胃癌肿瘤细胞株SGC-7901和肺癌肿瘤细胞株NCI-H460的抑制率在40%以上。【结论】药用植物南方红豆杉及其根际土壤蕴含种类丰富的放线菌资源,具有良好的生物学活性。菌株KLBMP 2170具有显著的抑菌以及抗肿瘤活性,值得我们去进一步研究。     

三种药用植物内生真菌抗真菌活性的研究

从雪上一枝蒿(Aconitumbrachypodum)、刺五加(Acanthopanaxsenticosus)和大血藤(Sargentodoxacuneata)分离获得的162株内生真菌进行抗真菌活性检测,结果具有抗菌活性菌株有74株,雪上一枝蒿15株,刺五加23株,大血藤36株,分别占3种植物各自内生真菌的44.12%,48.94%和44.44%。将具有抗菌活性的菌株进行分类鉴定,分属于6目、7科、23属。     

银杏内生菌的分离及其抑菌活性筛选

从健康的银杏（Ginkgo biloba）茎和叶片中分离内生菌,结果从银杏叶和茎上共分离到内生真菌20株,其中9株来自银杏茎部,11株来自银杏叶部;内生放线菌23株,其中15株来自于银杏茎部,8株来自于银杏叶部;内生细菌15株,其中8株来自于银杏茎部,7株来自于银杏叶部。以金黄色葡萄球菌（Staphylococcus aureus）、枯草芽胞杆菌（Bacillus subtilis）、大肠埃希菌（Escherichia coli）、黑曲霉（Aspergillus niger）、酿酒酵母（Saccharomyces cerevisiae）作为指示菌,采用双层平板法对内生菌进行抑菌活性筛选,结果表明：20株内生真菌中有12株出现了抑菌活性,有抑菌活性菌株的比例为60.0%;23株内生放线菌中,仅3株出现了抑菌活性,有抑菌活性菌株的比例为13.0%;15株内生细菌中,有4株出现了抑菌活性,有抑菌活性菌株的比例为26.7%。     

青海盐碱环境中具抗肿瘤活性放线菌的筛选和多样性研究

从我国青海省采集盐碱土样或泥样,用添加1.0~3.0mol/L NaCl的GPY琼脂培养基和ISP2琼脂培养基分离到145株典型放线菌菌株。采用6种肿瘤细胞株对分离菌株的发酵产物进行体外筛选,得到26株抗肿瘤活性阳性菌株(17.9%),19株为拟诺卡氏属(Nocardiopsis)菌株,7株为链霉菌属(Streptomyces)菌株。在抗肿瘤活性、形态特征、生理生化特性和全细胞水解物氨基酸组分分析等实验结果的基础上,选取差异较大的8株抗肿瘤活性阳性菌株进行16S rRNA基因序列测定和系统发育多样性分析。结果表明,2株属于链霉菌属(Streptomyces)的1个已知物种和1个潜在新种;6株属于拟诺卡氏菌属(Nocardiopsis),可能代表该属的4个新种。研究表明青海盐碱环境中存在产生抗肿瘤活性物质的重要放线菌资源,也提示其中蕴藏着较丰富的微生物多样性。     

Streptomyces sp. 173, an insecticidal micro-organism from marine

AIMS: To find new insecticidal antibiotics from marine micro-organisms. METHODS AND RESULTS: Strains isolated from seawater and sea sediments from Beidiahe and Dagang of the east coast of China were screened for their insecticidal qualities. The screening was carried out using bioassay of brine shrimp and the insect pest Helicoverpa armigera. The fermentation, preliminary extraction and isolation of Streptomyces sp.173 were carried out. CONCLUSIONS: In total 331 isolates were examined through bioassay of brine shrimp and 40 isolates (12.08%) showed potential insecticidal activities. Of the 40 isolates, one isolate, designated Streptomyces sp.173, was found to have strong insecticidal activity against both brine shrimp and H. armigera, similar to that of avermectin B1. SIGNIFICANCE AND IMPACT OF THE STUDY: The isolated Streptomyces sp.173 has great insecticidal potency. This work indicated that marine micro-organisms could be an important source of insecticidal antibiotics and the improved anti-brine shrimp bioassay is suitable for primary screening.     

Cristatumins A-D, new indole alkaloids from the marine-derived endophytic fungus Eurotium cristatum EN-220

Four new indole alkaloids, namely, cristatumins A-D (1-4), along with six known congeners (5-10) were identified from the culture extract of Eurotium cristatum EN-220, an endophytic fungus isolated from the marine alga Sargassum thunbergii. The structures of these compounds were established on the basis of extensive spectroscopic analysis. Each of these compounds was evaluated for antimicrobial and insecticidal activity. Compounds 1 and 9 showed antibacterial activity against Escherichia coli and Staphyloccocus aureus, respectively, while compounds 2, 6, and 7 exhibited moderate lethal activity against brine shrimp. Preliminary structure-activity relationships were also discussed.     

Polycyclic aromatic hydrocarbons and heavy metals in Kostrena coastal area

The aim of this study was to determine pollution by polycyclic aromatic hydrocarbons (PAH) and heavy metals in seawater and sediment in Kostrena coastal area, as well as their toxicity using bioluminescence based tests. Total PAH concentration in seawater ranged 1.7-155.3 ng/L. The share of carcinogenetic PAH was relatively high, ranging 22-48.3%. Nickel concentrations in seawater were beyond detection limits (< 0.1 microg/L), vanadium concentrations ranged 0.66-1.96 microg/L, chrome concentrations were beyond detection limits, and copper concentrations were also beyond detection limits or extremely low (up to 0.32 microg/L). EC50 values in seawater ranged 23.80-90.90 ng/L. Correlation between total PAH concentration and toxicity of seawater showed strong connection between them (r = 0.9579). Total PAH concentration in marine sediment ranged 58.02-1116 microg/kg dry weight (d.w.). The share of carcinogenetic PAH was extremely high ranging 10-53%. Nickel concentrations in marine sediment ranged 8-24 mg/kg d.w., vanadium concentrations ranged 24-42 mg/kg d.w., chrome concentrations ranged 11-19 mg/kg d.w., and copper concentrations ranged 7-25 mg/kg d.w. EC50 values in marine sediment ranged 818-4596 microg/kg d.w. Correlation between total PAH concentration and toxicity of marine sediment showed weak connection between them (r = 0.2590). Previous studies of seawater samples from areas of the Adriatic sea under the direct influence of oil industry did not include concentrations of heavy metals, which makes our study the first to present such comprehensive results. Our results point out the need for further evaluations and following of marine environment pollution and its consequences on living organisms and marine ecosystem in whole.     

产生物活性物质海洋放线菌的筛选和鉴定

本实验从辽宁大连海域的海水、海泥和海参养殖圈的样品中分离得到38株海洋放线菌,以金黄色葡萄球菌、溶壁微球菌、枯草芽孢杆菌、副溶血性弧菌和铜绿假单胞杆菌为指示菌,筛选出两株抑菌活性高的菌株,分别命名为HS-B31和HS-B34。16S r DNA测序鉴定及构建系统发育树的结果显示,它们都属于放线菌目、链霉菌属的不同种。对两株链霉菌的发酵上清液进行萃取、粗提和浓缩得到粗提物,即B31和B34;抗菌作用的结果显示,这两种粗提物的抗菌效果均为显著。经薄层层析分析,并利用制备型层析板对粗提物B31和B34进行活性物质的分离制备,共得到八个组分,即B31-1、B31-2、B31-3、B31-4、B31-5和B34-1、B34-2、B34-3。用滤纸片法对这些组分进行抗菌检测,结果显示组分B31-3和B34-3不仅对革兰氏阳性菌金黄色葡萄球菌有较好的抑菌作用,而且对海洋致病菌革兰氏阴性菌副溶血性弧菌也显示出较强的抑菌效果。研究为新型抗生素的研制和应用提供了微生物新资源。     

Inhibition of the development of microorganisms (bacteria and fungi) by extracts of marine algae from Brittany, France

The inhibitory effects of aqueous, ethanolic and dichloromethane fractions from 16 marine algae from the Atlantic shores of North-East Brittany, France, have been investigated against microorganisms frequently associated with immersed surfaces. The extracts were tested in vitro against isolates of marine fungi, bacteria and yeasts potentially involved at different stages in the formation of biofilms in the sea. The high levels of inhibitory activity of nine extracts against marine fungi and Gram-positive bacteria and their apparent absence of toxicity against larvae of oysters and sea urchins suggests a potential for novel active ingredients. Received: 30 December 1998 / Received revision: 25 April 2000 / Accepted: 1 May 2000     

Evidence for several types of biologically active substances in north pacific sea anemones

1. Twenty-two sea anemone samples from seven species were collected in Aleutian and Comandorskiye Islands from sub-littoral region (> 50 m depth).2. Water-ethanol extracts of sea anemones were tested using various test-systems after ethanol evaporation.3. All sea anemones extracts inhibited DNA and most of them inhibited RNA synthesis in Ehrlich carcinoma tumor cells.4. Extracts of most sea anemones species showed high hemolytic activity.5. The extracts proved to be nontoxic or display low toxicity being i.p. injected into mice.6. Some extracts precipitated virus of aleutian disease of mink.7. None of the extracts showed activity toward Gram + ve, Gram −ve bacteria or yeast.     

豆薯种子中的杀虫成分及其毒力测定

为明确豆薯Pachyrrhizus erosus (L.) Urban种子中的杀虫成分及其杀虫毒力,以白纹伊蚊Aedes albopictus 4龄幼虫为目标昆虫,在活性跟踪的基础上,通过多种色谱技术和核磁共振技术分离鉴定其化学成分结构,并通过浸渍法、点滴、夹毒叶碟法等测定各化合物对白纹伊蚊幼虫、棉蚜Aphis gossypii无翅成蚜、甘薯天蛾Herse convolvuli幼虫和小菜蛾Plutella xylostella幼虫的杀虫活性和作用方式。结果表明,从豆薯种子中共分离、鉴定了14个化合物,即12a-hydoxyrotenone,pachyrrhizine,12a-hydoxypachyrrhizone,12α-dehydropachyrrizone,α-naphthoflavone,7-methoxyflavanone,12α-hydroxydolineone,6-methoxyflavone,4′-hydoxyflavanone,quercetin dihydate,5-methoxyflavone,7-hydroxyflavone,3′-hydoxyflavanone和3-hydroxyflavone。上述化合物中的前7个化合物对白纹伊蚊4龄幼虫具有毒杀活性, 处理24 h的LC₅₀分别是25.0,51.1,196.2,48.4,98.9,107.2 和15.6 mg/L;前6个化合物对棉蚜成虫24 h 的毒力分别为1.5,10.9,80.7,8,32.1和112.8 mg/L;前5个化合物以及12α-hydroxydolineone对甘薯天蛾3龄幼虫有毒杀活性;12α-hydoxyrotenone对3龄小菜蛾幼虫24 h的胃毒毒力LD₅₀为17.3 μg/头,7-methoxyflavanone对该虫仅表现出较弱的生长发育抑制活性。首次从豆薯种子中分离得9个化合物,即6-methoxyflavone,4′-hydoxyflavanone,quercetin dihydate,α-naphthoflavone,7-methoxyflavanone,5-methoxyflavone,7-hydroxyflavone,3′-hydoxyflavanone和3-hydroxyflavone;豆薯种子含有7个活性成分,其作用方式因目标昆虫不同而不同,其主要杀虫成分不是鱼藤酮而是12α-羟基鱼藤酮。     

Microbial characteristics of marine sediments in bathing area along Pesaro-Gabicce coast (Italy): a preliminary study

AIMS: This paper evaluates the presence of human pathogen micro-organisms in marine sediments in a coastal area suitable for bathing. In addition, the grain size analysis was performed in order to correlate the characteristics of the sediments and the microbial content. METHODS AND RESULTS: The samples were collected in two small bays along the central Adriatic coast, where breakwaters had been built for the purpose of halting marine erosion. Faecal contamination indicators, Salmonella and Vibrio species, enteric viruses were investigated using standard techniques for isolation and identification. The grain size was determined using calibrated sieves and 'Sedigraph' device. In some samples, the faecal contamination indices overstepped legislative limits. Salmonella was never found. Vibrio and enteric viruses were isolated: the micro-organisms were preferentially abundant in fine sediments. CONCLUSIONs: Marine sediments can represent an important reservoir of allochthonous and marine micro-organisms and the microbial charge correlates with the characteristics of the sediments. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates that it is important to analyse marine sediments before defining the quality of coastal areas.     

Protease Formation by a Marine Psychrophilic Bacterium

Proteolytic activity was found in the culture fluids of numerous psychrophilic bacteria isolated from terrestrial or marine samples. Among these organisms, a marine psychrophilic bacterium, Pseudomonas sp. No. 548, showed the highest proteolytic activity. This organism required salts of sea water for both growth and protease formation. The optimum temperature for the growth of this organism was 20°C. The formation of protease was the greatest at 5°C and decreased with increasing temperature. The extracellular protease system was fractionated into at least two components having proteolytic activities by chromatography with DEAE-cellulose. Increasing culture temperature tended to increase the activity ratio of Fraction I to Fraction II. Some cultural conditions for protease formation were investigated.