Isolation of antifouling substances from the Palauan sponge Protophlitaspongia aga

ABSTRACT: Fouling organisms cause serious problems by settling on ships' hulls, and other marine infrastructures. Organotin compounds have been developed as effective antifoulants. However, environmental problems are associated with these compounds. It is necessary to find antifouling substances that exhibit a lower toxicity than those substances currently used. A new pyrimidine derivative, 3,4,5,6-tetrahydro-6-hydroxymethyl-3,6-dimethyl-4-pyrimidinecarboxylic acid, (compound 1) and zooanemonin (compound 2) were isolated from the marine sponge Protophlitaspongia aga as antifouling substances against the barnacle Balanus amphitrite . We also found that α-nicotinamide ribose (compound 3) inhibited germination and the attachment of Ulva spores.     

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.     

智利竹鱼渔场海洋微生物的物种多样性及其生物活性筛选

利用东海水产研究所开展的2012年东南太平洋智利竹鱼(Trachurus murphyi)渔场生态环境调查,对该海域75个站点进行了海洋微生物样品的采集;通过菌株的16S rRNA测序及GenBank数据库比对,分析了从所采样品中分离获得的海洋微生物菌株的物种多样性;通过菌株发酵、代谢产物提取及其抗菌滤纸扩散分析,对其发酵代谢产物粗提物进行了抗菌活性筛选;通过类胡萝卜素合成通路中八氢番茄红素合成酶(crtB)及脱氢酶(crtI)基因分析,对筛选到的抗菌活性菌株进行了基因筛选。结果表明,累计分离获得可培养海洋微生物菌株628株,其隶属12个属,其中的优势菌属包括亚硫酸盐杆菌属(Sulfitobacter)、弧菌属(Vibrios)及气单胞菌属(Aeromonas)。其中16株菌株的发酵代谢产物具有明显的抗菌活性,而其中12株活性菌株具有合成类胡萝卜素的crtB、crtI分子遗传基础。     

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.     

Progress,opportunities and challenges for marine conservation in the Pacific Islands

1. The people of the Pacific have long relied on the ocean for sustenance, commerce and cultural identity, which resulted in a sophisticated understanding of the marine environment and its conservation.
2. The global declines in ocean health require new and innovative approaches to conserving marine ecosystems. Marine Protected Areas (MPAs) have been shown to be a highly effective means of conserving biodiversity and managing fisheries, while also restoring and preserving overall ecosystem function.
3. Traditional ecological knowledge held by many island peoples in the Pacific is critical to the development, design and implementation of contemporary MPAs.
4. Chile's offshore islands are among the few oceanic archipelagos along the west coast of South America. These islands have cultural and ecological connections to the broader insular Pacific, yet our scientific understanding of them is extremely limited.
5. Chile has created several large-scale MPAs around their offshore archipelagos. By protecting these unique ecosystems, Chile has established itself as a global leader in marine conservation.
6. Effective management and a better understanding of social–ecological interactions are currently the biggest challenges facing MPAs in the Pacific Islands.

     

Geophysical approaches to the classification,delineation and monitoring of marine habitats and their communities

• 1. If marine environments are to be systematically protected from the adverse effects of human activities, then identification of the types of marine habitats and the communities they contain, and delineation of their boundaries utilizing a consistent classification is required. Human impacts on defined communities can then be assessed, the ‘health’ of these communities can be monitored, and marine protected areas can be designated as appropriate.
• 2. Schemes to classify habitats at local and regional scales, according to their geophysical properties, may identify different factors as determinants, and/or use them in different sequences in a hierarchical classification. We examined the reasons for these differences in local and regional applications of a global concept, and argue that a common set of factors could be applied in a defined and defensible sequence to produce a common hierarchy of habitat types among geographic regions.
• 3. We show how simple mapping and GIS techniques, based on readily available data, can lead to the identification of representative habitat types over broad geographic regions. We applied a geophysical framework first to the entire Canadian coastline and second to the Scotian Shelf of Atlantic Canada to establish broad scale marine natural regions and ‘seascapes’, respectively. This ecosystem level approach — which defines representative habitat types — is a fundamental prerequisite for many purposes. It can form the basis for further analyses including: definition of community types from habitat — community relationships; evaluation of the potential roles of focal species in marine conservation; evaluation of candidate marine protected areas; definition of unaffected reference areas against which the effects of human activities can be gauged; guidance for water quality monitoring studies; management of marine resources.

Copyright © 2003 John Wiley & Sons, Ltd.     

Characterization of a novel marine origin aerobic nitrifying–denitrifying bacterium isolated from shrimp culture ponds

A novel marine origin Bacillus subtilis strain H1 isolated from a shrimp culture pond effectively removed NH₄⁺‐N, ‐N and ‐N, with a maximum ammonium, nitrite and nitrate removal rate of 2.35 mg NH₄⁺‐N L^?1 hr^?1 per OD, 9.64 mg ‐N L^?1 hr^?1 per OD and 0.75 mg ‐N L^?1 hr^?1 respectively. The gas chromatography–isotope ratio mass spectrometry results indicated that N₂O was emitted when ¹⁵NH₄Cl, Na¹⁵NO₂ or Na¹⁵NO₃ was used. Additionally, N₂ was also produced when Na¹⁵NO₂ was used. Single‐factor experiments suggested that the optimal conditions for NH₄⁺‐N and ‐N removal were glucose as a carbon source, C/N 15, initial pH 7.5, 30 g/L NaCl, 28°C and a shaking speed of 160 rpm. Orthogonal tests showed that the optimal conditions for NH₄⁺‐N removal were C/N 15, pH 9, 10 g/L NaCl and shaking speed 160 rpm when ammonium chloride was used as the substrate. The optimal conditions for ‐N removal were C/N 10, pH 6, 10 g/L NaCl and a shaking speed of 160 rpm when sodium nitrite was used as the substrate. In summary, B. subtilis strain H1 had highly efficient aerobic nitrifying–denitrifying ability and high adaptability, suggesting that it is potentially valuable to marine aquaculture.     

Relationships between marine growth and marine survival of one sea winter Atlantic salmon, Salmo salar L., from the River Bush, Northern Ireland

An examination of marine growth/marine survival relationships in Atlantic salmon, Salmo salar L., was carried out, based on scale growth measurements in relation to two indices of marine survival in wild fish from the River Bush, Northern Ireland. The survival of cohorts to the Irish/Northern Irish coast (prefishery) and to fresh water was statistically unrelated to variation in growth from smolt migration to the end of the first winter at sea ( P  > 0.1; – P  > 0.7). Marine growth of 1+ smolts decreased significantly during the period of the study ( P  < 0.05), but growth of 2+ smolts did not change ( P  > 0.05). The variability in marine growth was much less than the variation in natural survival at sea, suggesting that factors instead of or in addition to, growth influence natural survival in the marine environment. Survival to fresh water was not related to survival to the coast ( P  > 0.4), although it was inversely correlated with exploitation rate ( P  < 0.01). These results are discussed in relation to the use of freshwater returns to assess marine survival and the potential for the variation in natural marine mortality to influence total life-cycle variation.     

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。     

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...     

Growth performance of bath sponge (Spongia officinalis Linnaeus, 1759) farmed on suspended ropes in the Dardanelles (Turkey)

Commercial bath sponge population in the Mediterranean has been recently reduced due to diseases and the depletion of natural banks. The commercial supply is far below the demand on the last 10–15 years. In this study, we aimed to investigate the farming performance of Spongia officinalis in the Dardanelles. Sponges collected from the Dardanelles cut into pieces of different weight. They were divided into two groups depending on the wet weight ranging from 50 to 150 g (mean weight 102.6 ± 4.81 g) and from 160 to 360 g (mean weight 235.8 ± 8.56 g). Three rope systems located parallel to each other with 6–7 m inter‐distance were constructed for the cultivation of sponges. At the end of the study, while small‐sized sponges could reach mean wet weight of 120.7 ± 14.8 g, large‐sized sponges reached mean wet weight of 247.6 ± 22.4 g. Growth rates were estimated 5–17% at 21st month. The survival rates were found 82% and 88%.     

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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You cannot conserve a species that has not been found: The case of the marine sponge Axinella polypoides in Liguria,Italy

1. Detailed knowledge about the distribution of species in need of protection is required for the management of Marine Protected Areas, a major tool to reduce marine biodiversity loss. Such knowledge is deficient for most marine invertebrates.
2. Axinella polypoides is a marine sponge included on the list of protected species by the Barcelona Convention (1976) and the Bern Convention (1987). This large and erect species has an important ecological role in habitat forming and benthic–pelagic coupling.
3. Bathymetrical, geographical and ecological data over the last 60 years were collated from publications and reports, together with new surveys to assess the distribution and protection status in Liguria of A. polypoides. It identified a more widespread distribution than previously thought, which points at a general need for dedicated investigations on the occurrence of species that require protection.
4. Bathymetrical distribution was trimodal, with peaks corresponding to different geomorphological settings: coastal cliff bases (around 38 m depth), inner shelf shoals (52 m) and rocks amidst coarse sediment on the outer shelf (79 m). Density was significantly greater at the shallowest depths. The species was mostly found in the coralligenous biocoenosis, in association with other characteristic species or forming a monospecific facies.
5. On (sub)vertical cliffs, A. polypoides often exhibited an unusual cane shape, rather than the typical bushy morphology, thus causing confusion with the congeneric A. cannabina, a more southern species. Records of the latter in the Ligurian Sea therefore need confirmation.
6. Only a minority (22.6%) of A. polypoides records were in Marine Protected Areas, the remainder being located in areas with no current environmental protection plans in place. While the occurrence of this species in MPAs remained stable over the decades, the only quantitative historical data available indicated that populations in non-protected areas were declining owing to anthropogenic impacts (fishing and anchoring).

     

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。     

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。     

Benthic foraminifera as proxies of marine influence in the Orosei marine caves,Sardinia, Italy

1. As a result of their location at the boundary between marine and continental domains, marine caves are affected by wide spatial and seasonal environmental changes. Only recently have benthic foraminifera been recognized as reliable indicators for the ecological zonation of these environments.
2. The present study is focused on two marine caves of the Orosei Gulf, Sardinia, Italy: Bue Marino and Bel Torrente. It investigates the spatial and seasonal variability of benthic foraminiferal assemblages relative to sediment grain size and water parameters (temperature, salinity, pH, and dissolved oxygen) collected during two campaigns in August 2014 and April 2015. The results from 2014 have been partially published.
3. Based on a comparison of the results of the two campaigns, the considerable reduction of foraminiferal abundance in Bel Torrente was deduced to occur because of the strong freshwater flows occurring during the rainy season; in Bue Marino, the less severe water flow allowed the identification of both living and dead foraminifera, although strongly reduced in number. These identifications allowed benthic foraminifera to be used to define the ecological zonation.
4. Entrance, confluence, and transitional ecozones were identified in Bue Marino cave on the basis of species abundance. The second ecozone, not recognized in 2014, was correlated with plant debris at the confluence of the two cave branches. The other two ecozones, which are characterized by the faunal shift from hyaline‐ to agglutinated‐prevalent assemblages, were attributed to the gradient of abiotic parameters detected from the outer to the inner portions of the cave. In both campaigns the same ecozones were recognized in terms of species composition, with exceptions being found to different extents as a result of seasonal variability.
5. As the distribution of foraminiferal ecozones is conditioned by a decreasing gradient of marine influence, long‐term monitoring may be regarded as a promising tool for future studies on sea‐level change.

     

寄生海水养殖鱼类的拟格拉夫涡虫的流行、危害与防治

自2000年10月中旬,在罗源湾发现一种罕见的能够寄生鱼类的涡虫,对其流行病学、形态学以及进化地位进行了大量的研究,结果表明：这种涡虫是一种未被报道过的新种,在福建省的各海水养殖区广泛流行,能够寄生于眼斑拟石首鱼（Sciaenops ocellatus）、大黄鱼（Pseudosciaena crocea）、鮸鱼（Miichthys miiuy）、牙鲆（Paralichthys olivaceus）、黑鲷（Sparus macrocephalus）和横纹东方鲍（Takifugu oblongus）等海水养殖鱼类的鳃、鳍及体表,造成寄生部位的严重损伤,鱼因呼吸困难或细菌的继发性感染而造成大量死亡,死亡率可达20％-60％;流行季节主要为秋季（8-11月）;水温22-26℃,盐度25-30。此外,提出了使用地下水,降低盐度、温度,合理用药等切实可行的防治本涡虫病的建议。     

海洋经济生物标本的采集、制作和保存

海洋经济生物标本是高校、科研院所从事相关海洋生物学教学与研究的重要材料。以藻类、鱼类、贝类、甲壳类等为例,提出标本采集前应明确的几个问题,阐述了海洋经济生物标本采集方法、制作流程、药品配制、科学保存和维护技术等。     

原子吸收法测定海洋生物体中Fe、Mn、Ni的方法研究

通过收集国内外现行标准、规范,以中国海洋领域现行标准体系为基础,研究以原子吸收法测定海洋生物体中Fe、Mn、Ni元素的标准方法。通过加入Cacl2消除火焰法测定Fe、Mn元素的干扰或加入Pd(NO3)2和Mg(NO3)2消除石墨炉法测定Mn、Ni元素的干扰;在特定温度以HNO3-H2O2常压消解或微波消解样品,Fe、Mn元素以火焰法测定、Mn、Ni元素以石墨炉法测定,实验结果表明Fe、Mn、Ni元素的溶出值均在标准值范围内,从而证明该方法具有可行性,适用于海洋生物体的标准方法。