Phenotypic characterization and cultivation conditions of inhibitor-producing fungus isolated from marine sediment

ABSTRACT:   Tyrosinase inhibitor-producing fungus of Trichoderma sp. strain H1-7, which was isolated from marine sediment, was investigated by phylogenetic analysis and physiological characteristics. Phylogenetic analyses of the strain were similar to Trichoderma atroviride or T. viride . Physiological characteristics of the strain were similar to T. viride , and based on these results, it was identified as T. viride . Characterization of tyrosinase inhibitory activity of a culture supernatant of the strain was investigated. The inhibitory activity of the supernatant of the strain decreased after cultivation for more than 3 days. Furthermore, sea water was not essential for the production of the tyrosinase inhibitor (TI). When TI production of the strain was compared to T. viride , the strain showed higher activity than T. viride . From this result, it seemed that the strain had characteristic features comparable to T. viride , which was isolated from the terrestrial environment. As TI production of the strain showed higher potential than that of T. viride , it is necessary to elucidate the chemical structure of TI exemplified in the present study.     

海洋微生物活性物质的研究

海洋微生物由于其生存环境的特殊性,发展出独特的代谢方式,由此产生了结构新颖,功能多样的活性物质。综述了抗菌、抗肿瘤、酶、酶抑制剂、海洋生物毒素、不饱和脂肪酸等活性物质的研究进展,展现出海洋活性物质的巨大开发潜力。     

一株产吡咯喹啉醌南极海洋细菌的分离鉴定及生物活性

从采集自南极FAO 48.1海域海水样品中分离纯化到一株产吡咯喹啉醌(PQQ)海洋细菌,对其进行了细菌分类学鉴定、摇瓶培养发酵、发酵产物提取及HPLC定量分析,并对纯化的PQQ进行了针对一氧化氮(NO)抑制活性及羟自由基(.OH)清除抗氧化活性评价。结果表明,该菌株与副球菌属(Paracoccus sp.)内菌株的16S rRNA基因序列同源性较高。综合多个分类学指标的分析结果,将其鉴定为属于副球菌属,命名为Paracoccus sp.PQ-08。生物活性评价结果表明,PQQ可显著抑制人脐静脉内皮(ECV-304)模式细胞内诱导型一氧化氮合成酶(iNOS)在mRNA转录水平及蛋白水平的表达量,明显降低胞内的NO释放量。当其作用浓度为100μM时,对iNOS蛋白表达的最高抑制率为67.9%(P<0.05)。此外,实验还证实PQQ具有较强的清除.OH的抗氧化活性。     

基于DNA条形码技术对江门沿岸海域夏季鱼卵的鉴定

以江门沿岸海域夏季采集到的鱼卵研究对象,利用DNA条形码技术分析鉴定鱼卵种类。本研究获得鱼卵个体有效线粒体COI序列信息217个,测序成功率为68.5%。经BOLD数据库比对分析,成功鉴定鱼卵5目14科19属20种(未知种2种);种内遗传距离为0~0.006,平均遗传距离为0.002,种间遗传距离为0.149~0.325,平均遗传距离为0.255,种间遗传距离为种内遗传的128倍;鱼卵样品以鲈形目数量最多,占51.8%;鲱形目次之,占24.8%;其中鱼卵优势种为黄斑鲾(Photopectoralis bindus)、日本鳀(Engraulis japonicus)、大甲鲹(Megalaspis cordyla)、粗鳞鮻(Chelon subviridis)、金钱鱼(Scatophagus argus)、龙头鱼(Harpadon nehereus)、亚洲(Sillago asiatica)。本次调查川山群岛东部海域获得鱼卵数量、鱼卵种类数均最多,是江门沿岸海域日本鳀、龙头鱼的主要产卵场;黄茅海、镇海湾西部水域均未获得鱼卵,可能与水域环境变化及陆源污染导致产卵环境破坏、亲体量减少有关。该研究结果揭示了江门沿岸海域夏季鱼卵分布格局,为其制定有效的产卵场保护措施提供依据,同时表明DNA条形码技术能有效地对鱼卵进行种类鉴定,可被广泛地应用于我国沿海海域鱼卵鉴定工作。     

几种海藻中溴过氧化物酶的筛选及酶学性质

溴过氧化物酶(BrPOD)是具有特殊功能的过氧化物酶,海藻是其主要来源。对几种中国海域的红藻如角叉菜(Chondrus ocellatus)、龙须菜(Gracilaria sjoestedtii)、珊瑚藻(Corallina officinalis)进行了溴过氧化物酶的藻种筛选,并对酶活较高的珊瑚藻进行溴过氧化物酶分离纯化及性质的研究。通过硫酸铵沉淀、DEAE-cellulose 52离子交换层析、Sephadex G-100凝胶层析等方法,从珊瑚藻中分离得到溴过氧化物酶。对该酶性质研究表明,该酶分子量较大,表观分子量为64 kD;溴化单氯甲酮的最适pH值为6.0;pH在5.0-9.0时酶活性稳定;在30-70℃温度范围内酶活性稳定;Ca2 、Co2 、Cu2 、Mn2 、NaF和EDTA等化合物使溴过氧化物酶活性下降,钒酸盐能提高酶活性。反应动力学实验表明,该酶对Br-、H2O2的Km分别为7.40 mmol/L和96.09μmol/L。     

2株海水鱼源潜在益生菌的分离、鉴定及特性分析

为筛选用于微生态制剂研发的海水鱼源益生菌,本研究对海捕野生许氏平鲉(Sebastes schlegelii)和大泷六线鱼(Hexagrammos otakii)的消化道内壁黏膜样品进行细菌分离纯化,获得80株可培养细菌。使用选择性培养基对菌株的产酶能力进行测定,选取海水鱼常见病原菌为指示菌测定分离菌株产物的抑菌活性,筛选出2株潜在益生菌TS2和TH8,并进行菌株的生理生化检测、16S rDNA序列分析、生长特性及其对宿主安全性的研究。结果显示,TS2产蛋白酶(protease)、淀粉酶(amylase)和脂肪酶(lipase),其无菌培养产物可显著抑制鳗弧菌(Vibrio anguillarum)、副溶血弧菌(Vibrio parahaemolyticus)、哈维氏弧菌(Vibrio harvey)和假交替单胞菌(Pseudoalteromonas nigrifaciens)的生长。TH8产蛋白酶和脂肪酶,其无菌培养产物可显著抑制鳗弧菌、溶藻弧菌(Vibrio alginolyticus)、副溶血弧菌、假交替单胞菌、嗜水气单胞菌(Aeromonas hydrophila)、金黄色葡萄球...     

多毛类沙蚕在沿岸海洋污染环境中的生物修复作用

随着经济社会的快速发展,人类对海洋生态环境的负面效应逐渐显现出来,海洋污染环境影响了人类的正常生活.为了防止沿岸海洋环境污染和海洋生态环境的持续恶化,各国都采取积极的响应措施.为了减少有害污染物和避免生态环境恶化,国内外的许多专家和学者提出利用多毛类的生物修复能力来有效地缓解沿岸海洋环境污染的压力.     

Characterizing regime shifts in the marine environment

Recent years have seen a plethora of studies reporting that ‘regime shifts’ have occurred in the North Atlantic and Pacific Oceans during the last century. In many cases, the criteria used to distinguish a regime shift have not been explicitly stated. In other cases, a formal definition has been proposed and the data set assessed against it. Developing a universal quantitative definition for identifying and distinguishing between purported climatic and ecological regime shifts has proved problematic as many authors have developed criteria that seem unique to the system under study. Consequently, they throw little light on the drivers of ecological regime shifts. Criteria used to define regime shifts are reviewed and on the basis of evidence from purported regime shifts, common characteristics in the speed and amplitude of the changes and the duration of quasi‐stable states are used to propose a more clearly defined set of criteria for defining climatic and ecological regime shifts. Causal drivers of regime shifts are explored using correlation analysis. Limitations of these methods are discussed.     

海州湾海洋牧场水环境的承载力

海洋牧场是实现海洋环境保护与渔业资源养护的重要举措。本研究以2014年春(5月)、夏(8月)、秋(10月)对海州湾海洋牧场示范区水环境数据为基础,选取高锰酸盐指数(CODMn)、溶解无机氮(DIN)、溶解无机磷(DIP)、生化需氧量(BOD)作为评价指标,利用BP神经网络模型对该海域水环境承载力进行评价。结果显示,2014年海州湾海洋牧场水环境承载力指数平均值高于0.6,承载状态较理想。水环境承载力存在明显的季节变化,呈现出夏季春季秋季;海洋牧场区域水环境承载力优于对照区海域;CODMn、DIN浓度过高是导致部分站点轻度超载的主要原因,这可能与陆源污染有关。研究表明,航运对水环境承载状态有负面影响;BP神经网络模型构建方便快捷,评价结果客观合理,可应用于海洋牧场等海域水环境承载力的研究。     

海洋微生物酶的研究概况

海洋中蕴藏着丰富的微生物资源,海洋微生物对于维持海洋生态平衡发挥着重要作用。尽管海洋中的许多微生物推测是由陆地环境经河水、污水、雨水或尘埃等途径而来的,但特殊的海洋环境赋予海洋微生物以特殊的遗传结构和生活习性。海洋生态环境复杂,高盐度、高压力、低温及特殊的光     

Induction and characterization of a lysogenic bacteriophage of Lactococcus garvieae isolated from marine fish species

This study investigated the presence of prophages in Lactococcus garvieae isolated from several marine fish species in Japan. Representative strains of 16 bacterial genotypes (S1–S16) selected from more than 400 L. garvieae isolates were used to induce lysogenic bacteriophages. These strains were treated with 500 ng mL^?1 freshly prepared mitomycin C. A cross‐spotting assay was performed to validate the lysogenic and indicator strains. The lysogenic strains were selected for isolation and concentration of the phages. Phage DNA was digested with EcoRI for biased sinusoidal field gel electrophoresis analysis. Polymerase chain reaction (PCR) was used to detect integrated prophage DNA. Of the 16 representative bacterial genotypes, 12 strains integrated prophages as indicated by the PCR assay, and 10 phages were detected and isolated using two indicator bacterial strains. Analysis of genomic DNA showed that these phages were homologous and named as PLgT‐1. Transmission electron microscopy revealed that the morphology of PLgT‐1 was consistent with the virus family Siphoviridae. PCR analysis of the prophage DNA revealed that all of the S1 genotype strains were lysogenic (30/30), but none of the S16 genotype strains were lysogenic (0/30). This is the first study to investigate lysogenic bacteriophages from L. garvieae.     

Isolation of a novel strain of Bacillus pumilus from penaeid shrimp that is inhibitory against marine pathogens.

A bacterium was isolated from the mid-gut of healthy black tiger shrimp, Penaeus monodon , based on a large zone of inhibition in mixed culture on solid medium. The isolate was a Gram-positive, motile spore former, with an optimum pH range for growth in tryptone soya broth containing 2% NaCl of between pH 6 and 9. The bacterium was highly salt tolerant with concentrations between 0% and 8% having no detrimental effect on growth. The isolate was identified as Bacillus pumilus based on physiological capabilities using the API50CHB and Biolog systems. Amplification and sequencing of the 16S rRNA gene followed by phylogenetic analysis confirmed its identity. The Bacillus pumilus isolate was strongly inhibitory against the marine bacterial pathogens Vibrio alginolyticus , V. mimicus and V. harveyi , and weakly inhibitory against V. parahaemolyticus in cross-streaking assays on solid medium. The organism was marginally self-inhibitory, and inhibited B. licheniformis and B. subtilis . The suitability of the B. pumilus isolate for use as a probiotic in farmed shrimp was further supported by the absence of any of the known B. cereus enterotoxin genes. Based on these in vitro results, in vivo safety and efficacy trials are underway to determine suitability of the novel strain as a commercial probiotic.     

Food habits and marine survival of juvenile Chinook and coho salmon from marine waters of Southeast Alaska

Little is known about the food habits of juvenile Chinook (Oncorhynchus tshawytscha) and coho (Oncorhynchus kisutch) salmon in marine environments of Alaska, or whether their diets may have contributed to extremely high marine survival rates for coho salmon from Southeast Alaska and much more modest survival rates for Southeast Alaskan Chinook salmon. To address these issues, we documented the spatial and temporal variability of diets of both species collected from marine waters of Southeast Alaska during summers of 1997–2000. Food habits were similar: major prey items of both species included fishes, crab larvae, hyperiid amphipods, insects, and euphausiids. Multivariate analyses of diet composition indicated that the most distinct groups were formed at the smallest spatial and temporal scales (the haul), although groups also formed at larger scales, such as by month or habitat type. Our expectations for how food habits would influence survival were only partially supported. As predicted, Southeast Alaskan coho salmon had more prey in their stomachs overall [1.8% of body weight (BW)] and proportionally far fewer empty stomachs (0.7%) than either Alaskan Chinook (1.4% BW, 5.1% empty) or coho salmon from other regions. However, contrary to our expectations, coho salmon diets contained surprisingly few fish (49% by weight). Apparently, Alaskan coho salmon achieved extremely high marine survival rates despite a diet consisting largely of small, less energetically‐efficient crustacean prey. Our results suggest that diet quantity (how much is eaten) rather than diet quality (what is eaten) is important to marine survival.     

海洋渔场生态环境质量状况综合评价方法探讨

海洋渔场生态环境质量状况的评价与保护是海洋渔业资源和渔业生态环境的基础。西方从海洋水文、海洋化学、初级生产力水平和饵料生物水平等方面对其进行综合性探讨，初步建立了海洋渔场生态环境质量综合指数评价方法，并以北部湾渔场为例进行评价实践，旨为建立一套较成熟的综合评价方法，从而为我国海洋渔业生态环境评价提供参考依据。     

Size and age are important factors for marine sanctuaries: evidence from a decade of systematic sampling in a subtropical marine park

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海洋侧孢短芽孢杆菌S-12-86溶菌酶的分离纯化与冻干技术研究

海洋微生物溶菌酶发酵液经低温离心、超滤浓缩、乙醇提取、Superdex 75 10/300凝胶层析和反相高效液相色谱层析纯化得到电泳纯海洋溶菌酶,分子量为17.1 kD,比活达到3 987.7 U/mg,纯度提高41.98倍,活力回收率为21.7%。对该酶冷冻干燥过程的研究结果表明,海藻糖、蔗糖和麦芽糖对该酶均有一定的冻干保护作用。其中,以海藻糖的保护效果最佳。0.5 mol/L海藻糖与20 mg/ml吐温80复合作为冻干保护剂,使冻干后的溶菌酶活性维持在95%以上,为该酶进一步研究和应用提供了稳定的酶制剂。     

Isolation and identification of indigenous marine diatoms (Bacillariophyta) for biomass production in open raceway ponds

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土壤中单宁和植酸降解菌的筛选、鉴定及液态发酵研究

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致病性哈维氏弧菌溶血素基因克隆及其检测

从山东沿海的发病鲈鱼（Lateolabrax japonicus）分离到1株致病性哈维氏弧菌（Vibrio harveyi），从该菌的染色体DNA扩增出一条长约1．4kb的特异性片段。DNA序列分析表明，该克隆片段含有完整的1254bp溶血素基因，该溶血素基因与哈维氏弧菌VIB645的溶血素基因VhhA和VhhB的相似性分别为99．0％和98．5％；与副溶血弧菌（V．parahaemolyticus）热稳定性溶血素基因（TDH）的相似性为74．5％；与拟态弧菌（V．mimicus）、创伤弧菌（V．vulnificus）、霍乱弧菌（V．chderae）磷脂酶基因的序列相似性分别为57．4％、59．2％、53．0％；与最小弧菌、创伤弧菌、霍乱弧菌、霍氏弧菌（V．hollisae）、河流弧菌（V．tguvialis）、鳗弧菌（V．anguillarum）的溶血素基因的相似性仅为19．9％～24．8％。根据溶血素基因的保守区段，设计了1对特异引物。分析表明，该引物能特异检测哈维氏弧菌。其可检测的DNA最小量为0．001ng。用该方法对55株不同来源的患病鱼分离的疑似病原菌进行检测，结果检出8株哈维氏弧菌，检出率为14．55％，从健康动物中检出数占所检弧菌数的6．25％，海洋环境为10．53％，海水养殖环境为26．53％，表明哈维氏弧菌在不同的海水环境及健康海洋动物中普遍存在，在养殖水体中的数量高于其他海水环境。     

海洋遥感技术在海洋渔业及相关领域的应用与研究

自20世纪70年代始,科研人员将遥感技术应用于海洋渔业及其相关领域的研究,将各类卫星遥感所获得的数据对海洋水温、海流、光、盐度、溶解氧、气象因素、水深、海底地形、饵料生物等进行了由定性到定量的分析,并将所得结果用于指导渔业生产、促进渔业研究、预警海洋灾害等方面,所起作用非传统调查方法能为。因此,遥感技术在海洋渔业及其相关领域正在发挥着越来越大的作用,并预示着广阔的应用前景和巨大的应用潜力。本文综述了遥感技术在海洋生态系统相关要素和渔场变动、大型动植物分布、海况监测、海洋生态污染及灾害监测等方面应用与研究进展,并对海洋遥感信息在我国海洋渔业领域的应用研究前景进行了分析。