Nitrification and the enumeration of marine nitrifying bacteria in a closed prawn-culture system

Nitrification in a closed prawn-culture system was well established and followed the classical oxidation sequence of ammonia via nitrite to nitrate. Ammonia levels were in the range 4,8–68,7 μg-at N·?^?1 nitrite 0,15–891 μg-at N·?^?1 and nitrate approached a maximum of 9 898 μg-at N·?^?1 after 22 weeks. Marine nitrifying bacteria were enumerated by the most probable number (MPN) technique. The maximum MPN estimate of either group of nitrifying bacteria in the filter was 1,73 × 10⁷·cm^?3 while the maximum MPN estimate of their counterparts in the culture water was approximately 500 times lower. To establish mean incubation times for the accurate enumeration of nitrifying bacteria, incubations were carried out over a period of 130 days. Maximum estimates of MPN of ammonia-oxidizing bacteria were obtained after an incubation period of 20 days with a mean of 15 days. Nitrite-oxidizing bacteria required a maximum of 65 days with a mean incubation period of 30,3 days.     

活石（live rock）在珊瑚养殖体系中对NH4+、NO3-的去除作用

Maintaining stable water quality is one of the key processes for recirculating coral aquaculture. Traditional aquarium systems which mainly utilized a nitrification of nitrifying bacteria attached to the surface of massive artificial filter material are difficult to maintain the oligotrophic conditions necessary for coral aquaculture. This study investigated the removal effects of dissolved inorganic nitrogen(ammonia and nitrate) by live rock(LR), a key component in the "Berlin system" coral aquarium. The expression levels of bacterial functional genes, AOA3,amo A and nos Z, were measured on the exterior and interior of LR. The nitrifying and denitrifying bacterial abundance on LR was quantified and the nitrogen nutrient regulatory effects of LR were evaluated. The results demonstrated that LR mainly removed ammonium(NH_4~+) from the water with a mean efficiency of 0.141 mg/(kg·h), while the removal of nitrate(NO_3~–) was not significant. Bacterial diversity analysis showed that ammonia-oxidizing bacteria(AOB) were the most common bacteria on LR, which accounted for 0.5%–1.4% of the total bacterial population, followed by denitrifying bacteria, which accounted for 0.2% of the total population, and the ammonia-oxidizing archaea(AOA) were the least common type(0.01%). The low abundance of denitrifying bacteria may be responsible for the poor nitrate(NO_3~–) removal of LR. Thus, other biological filtration methods are needed in coral aquaria to control nitrates generated from nitrification or biological metabolism.     

海洋微藻除菌及除菌与自然带菌微藻生长特点比较

于1995年4—12月在中国科学院海洋研究所进行微藻除菌及比较除菌与自然带菌微藻生长特点的研究。经平板培养排除霉菌后,利用组合抗生素(青霉素+卡那霉素+链霉素+庆大霉素)获得除菌球等鞭金藻、三角褐指藻及小球藻,对抗生素处理前后的微藻生长特点进行比较研究。结果表明,与未除菌时相比,除菌后的球等鞭金藻及小球藻不易老化(可保持良好悬浮30d以上);回加细菌于除菌藻,藻细胞下沉附底,说明细菌可促使微藻细胞老化。无维生素时,除菌后的球等鞭金藻生长更差,暗示未除菌的球等鞭金藻培养液内可能存在产(类)维生素细菌。除菌后三角褐指藻细胞形态发生一定变化,回加细菌后藻细胞形态有部分恢复。与未除菌时相比,除菌后三角褐指藻更能耐受高温(如30℃)。另外,某些抗生素能够刺激球等鞭金藻的生长。     

近海沉积物与水体中天然大分子聚合物降解菌的原位富集与多样性分析

本研究选取5种海洋动植物大分子高聚物或其天然组织,包括虾壳、鱼鳞、海带叶片、几丁质和壳聚糖,分别在海水表层和沉积物环境中进行富集,定期取样,通过高通量测序分析菌群多样性。结果发现,不同有机物原位富集的细菌多样性存在较大的差异,而且同种底物在海水表层与沉积物中的降解菌菌群差异较大。从物种多样性看,在海水表层环境中富集的鱼鳞样品种群最丰富,而沉积物环境中富集的海带叶片样品菌群多样性最低,除其优势菌群为热袍菌门外,其他所有富集物中优势菌均为变形菌门。其中脱硫杆菌科、黄杆菌科、脱硫弧菌科和弧菌科占有较大比例,脱硫杆菌科在所有样品中优势较大,黄杆菌科在海水表层环境样品中为优势菌群,弧菌科在沉积物样品几丁质和壳聚糖样品中占比较高。本研究通过对同种海洋环境中不同富集样品之间以及同种底物不同环境中富集菌群之间的比较,分析结果得到了原位条件下参与大分子聚合物降解的菌群种类,但有待于在更多不同的海域富集物中进行验证。     

源自生物絮团产絮凝剂的异养硝化-好氧反硝化菌xt1的鉴定及其脱氮特性

为了高效进行水体脱氮,本实验从形成于凡纳滨对虾(Litopenaeus vannamei)养殖水体的生物絮团中分离到一株具产絮能力的脱氮菌xt1,经16S r RNA基因测序与生理生化分析确定菌株xt1为短小芽孢杆菌(Bacillus pumilus)。在此基础上,本文研究了该菌的脱氮特性。结果表明:菌株xt1最佳碳源为葡萄糖,以其为底物对氨氮、硝态氮去除率分别达95.56%和57.40%。以蔗糖为碳源亦具较高脱氮率,对氨氮、硝态氮去除率分别达69.95%和49.50%;该菌能利用有机氮加速生长,添加0.25%、0.5%、1%和2%的蛋白胨能促进OD600,分别达到0.925、1.034、1.103和1.314,均高于未加蛋白胨下的生长,且氨氮去除率均超过90%,硝态氮去除率均超过88%;该菌能适应20—200mg/L无机氮浓度;该菌能以NH4+-N、NO2–-N或NO3–-N为唯一氮源进行异养硝化-好氧反硝化,反应84h去除率分别达到94.16%、47.60%和91.17%。其中,该菌的硝化形式是将氨氮转化为气态氮脱除,其硝态氮反硝化形式是先将硝态氮转化为亚硝氮,再以气态氮脱除。在进行异养硝化-好氧反硝化同时,菌株xt1体现絮凝特性,絮凝率最高分别达到82.28%、73.15%和75.60%;此外,添加该菌于养殖水体中能加速生物絮团形成,同时提高脱氮率。各项结果表明,菌株xt1可作为水产养殖水体脱氮的备选菌株。     

Molecular diversity of Thermococcales isolated from Guaymas Basin hydrothermal vents

Members of Thermococcales are organoheterotrophic hyperthermophilic anaerobes consisting of three genera, Thermococcus, Pyrococcus, and Palaeococcus, among which Thermococcus are the most frequently isolated. In this study, a variety of Thermococcales strains were isolated from the Guaymas Basin hydrothermal vents under different temperatures and pressures. Based on their 16S rRNA gene sequences, all the strains isolated from 85 C or 95 C enrichment cultures at ambient pressure were classified as Thermococcus, while strains isolated from 108 C-30 MPa enrichment cultures belonged to Pyrococcus. The combination of high pressure and high temperature for enrichment was shown to be an efficient method for isolating Pyrococcus strains. Through selecting and analyzing with multiple molecular phylogenetic markers, the isolated Thermococcales strains were found quite diversified including several putative novel species. This demonstrates a vast genetic reservoir of Thermoccoccales in the deep-sea hydrothermal vents, and a fast evolving of the Thermococcales species adapting to the fast changing environment.     

Toxicological effects of potash brine on bay of Fundy marine organisms

The acute lethality and sublethal heavy metal contamination potential to marine organisms of a waste brine solution from a potash mining operation were determined. The brine was acutely lethal to marine organisms of the Bay of Fundy, Canada, at concentrations between 47 and 55‰ salinity (96-h LC₅₀ values: threespine stickleback, Gasterosteus aculeatus, 47·7‰; polychaete worm, Nephtys incisa, 52·5‰; blue mussel, Mytilus edulis, lethal to whole animal and reduced ciliary activity of gills above 55‰).The brine diluted to sublethal concentrations (≤40‰) contained manganese (≤ 49·5 ×), copper and zinc (≤ 5·2 ×), lead (≤ 2·6 ×) and cadmium (≤ 0·7 ×) in decreasing order of enrichment relative to control seawater. Copper was accumulated in mussels exposed to these solutions over 44 days to the greatest degree (4 ·4 ×), followed by manganese and lead. Tissue lead concentration reached a steady value within 44 days and cadmium was not accumulated.     

不同碳源富集的石油烃降解菌群结构的分析

以原油和正十六烷为唯一碳源,从长期受石油污染的环境中富集和分离具有不同功能的石油烃降解菌,并利用平板法和PCR-DGGE法时不同碳源富集到的菌群结构进行分析.结果表明,利用2216E平板从以原油和正十六烷为碳源的富集液中各得到两株菌,分另11为TJ-1、TJ-2和TJL-1和TJL-2,分子生物学方法鉴定结果表明,这4株菌分别为短小芽孢杆菌(Bacillus pumilus)、威尼斯不动杆菌(Acinetobacter venetianus)、Oceanobacillus picturaeP和短小芽孢杆菌(Bacillus pumilus).PCR-DGGE分析结果表明,以原油和正十六烷为碳源的富集液中优势菌分别有5种和2种,且2种富集液中的优势菌明显不同.对比PCR-DGGE和平板法分析结果,可以看出PCR-DGGE法能够提供更全面的菌群结构信息.     

重金属锌对海洋微藻藻源氨基酸产出的影响

锌是海洋微藻生长必需的金属元素,低浓度时可促进微藻的生长,而在高浓度时则表现为抑制其生长。受人类活动的影响,海洋中锌有富集增加的趋势,从而会对海洋微藻的生长产生影响。本论文探究了锌离子以及纳米氧化锌对硅藻中肋骨条藻(Skeletonemacostatum)和新月菱形藻(Nitzschia closterium)生长的影响,特别是对藻源氨基酸产生的影响,结果发现:纳米金属颗粒会通过释放离子的形式抑制海洋浮游植物的生长,中肋骨条藻比新月菱形藻对锌的响应更敏感。锌离子和纳米氧化锌可通过抑制浮游植物的生长来减少氨基酸的释放量和改变藻源氨基酸的种类,从而可能会对海洋氮循环过程产生影响。     

海洋异养细菌对小球藻生长和无机营养盐吸收的影响研究

从胶州湾海洋异养细菌同小球藻(Chlorella vulgaris)的共存体系中,分离出6株形态特征差异较大的异养细菌,并做变性凝胶梯度电泳(Denature Gel Gradient Electrophoresis,DGGE)分析.其中一株对小球藻的生长有较大的促进作用,而其余菌株则没有明显的效果.6株细菌能在小球藻产生的胞外溶解有机物(Dissolved Organic Mate rials,DOM)溶液中生长良好,共存细菌对提高菌藻体系对无机营养盐的吸收具有较大的贡献.     

Dietary requirements of seaweed flies (Coelopa frigida)

The seaweed fly, Coelopa frigida (Fabricius), is mostly found in piles of decomposing seaweed deposited on the seashore which form its only breeding sites. It is shown that C. frigida can complete its life cycle in a wide variety of marine algae, and that the larvae are unable to survive without some, as yet unidentified, consituent of seaweed. The larvae also have a requirement for a microbial gut flora which probably derives from the bacterial flora naturally associated with algae growing in the sea. After deposition of the seaweed on the shore, the bacterial population increases enormously, and is ingested by the feeding Coelopa larvae. The dietary requirement for bacteria can be satisfied by a variety of pure bacterial cultures of marine origin, and also by pure cultures of Escherichia coli, Bacillus subtilis and Saccharomyces cerevisiae. It is suggested that the microbial cells are being used by the larvae as their principal source of energy. The bacterial populations naturally found on stranded seaweed are grazed by the feeding larvae. It is the combined activities of microbial and insect populations that result in rapid decomposition of the seaweed. The ecological relationships between marine algae, the microbial flora, and dipteran larvae are discussed.     

钻井液特征污染物的毒性评估和海洋测试生物的适用性比较

随着海上钻井数量的逐年增加、钻井液的使用量迅速增加,与之相关的环境危害引起了广泛关注,因此有必要筛选合适的毒性测试生物和开发环境友好的钻井液。本研究中,按照ISO11348-1（2007）、GB/T21805-2008和GB/T18420.2-2009的毒性测试方法,采用标准的海洋测试生物：发光细菌（Vibrio fischeri）、牟氏角毛藻（Chaetoceros muelleri）、诸氏鲻虾虎鱼（Mugilogobius chulae）、凡纳滨对虾（Litopenaeus vannamei）、卤虫（Artemia）、蒙古裸腹溞（Moina mongolica）,研究了钻井废液特征污染物（苯酚、七水硫酸锌、氯化汞）以及参比毒物十二烷基硫酸钠（Sodium dodecyl sulfate,SDS）的急性毒性效应。结果表明,钻井液特征污染物对6种海洋生物的急性毒性效应有所不同：Hg²⁺对海洋生物表现为极高毒性,Zn²⁺、SDS表现为中、高毒性,苯酚表现为中、低毒性;6种海洋生物对4种特征污染物的敏感性顺序为：蒙古裸腹溞 > 诸氏鲻虾虎鱼 > 卤虫 > 凡纳滨对虾 > 发光细菌 > 牟氏角毛藻;诸氏鲻虾虎鱼与卤虫、凡纳滨对虾种间毒性相关性显著,蒙古裸腹溞与发光细菌、凡纳滨对虾种间毒性相关性显著。上述研究结果为环保型海洋钻井液开发和钻井废液生物毒性评价中受试生物的选择提供了基础数据。     

南、北极近岸海水中好氧不产氧光合基因pufM的多样性

好氧不产氧光合(AAP)细菌因其具有利用溶解性有机物及光能的能力,在海洋碳循环及能量流动中发挥着重要的作用。该类细菌广泛分布在海洋环境中,其在不同生境中的多样性已被调查。但到目前为止,人们对于高纬度地区好氧不产氧光合细菌的认识还较为缺乏。有鉴于此,本研究基于编码光反应复合物上一个色素结合蛋白亚基的pufM基因,对北极王湾及南极乔治王岛近岸水体中夏季好氧不产氧光合细菌的多样性进行了检测。针对2个王湾站位和2个南极麦克斯维尔湾站位构建了4个pufM基因克隆文库,获得674个阳性克隆子。北极克隆子全部由α-变形细菌组成,而南极克隆子则包括α-变形细菌及β-变形细菌。来源于类似红细菌科的pufM基因在所有样品中皆占据优势。此外,与一株滴状亚硫酸盐杆菌中质粒编码的pufM基因存在亲缘关系的序列,在南、北极样品中均占优势。结果表明海洋环境中的pufM基因存在跨极甚至是环球分布。与此同时,南、北极序列之间的差异,也表明了极地地方种的存在。这些结果显示,作为好氧不产氧光合细菌的红细菌科在两极的近岸水体中具有重要的地位。     

乐清湾大型底栖动物群落及其与环境因子之间的关系

为了解和掌握乐清湾大型底栖动物群落及其与环境因子的相关性,于2015年1月、3月、5月、8月和10月开展了5个航次的大型底栖动物和环境因子的调查。结果表明,1月、3月和5月3个月份的群落结构和空间分布均较为接近,优势群落的主要特征种为薄云母蛤和不倒翁虫;8月和10月的优势群落不明显,主要群落特征种有不倒翁虫、双形拟单指虫、寡鳃齿吻沙蚕、小头虫、中蚓虫属一种等。丰度/生物量曲线(ABC曲线)分析表明5月的群落结构较稳定,1月、3月、8月和10月均处于不同程度的扰动状态,其中8月的受扰动程度最大。BVSTEP分析表明亚硝酸盐是乐清湾大型底栖动物群落各月份差异的主要环境驱动因子,盐度和氨氮分别是影响乐清湾3月和5月大型底栖动物群落结构的主要环境因子。典范对应分析(CCA)表明水温对乐清湾大型底栖群落特征种时空分布影响最大,其次是盐度、溶解氧、含氮营养盐和pH等。通过CCA排序图发现大部分群落特征种分布在含氮营养盐较低的水域,表明高浓度的含氮营养盐已经对乐清湾的大型底栖动物产生了负面影响。     

芽孢杆菌(Bacillus)及其复合粘土对典型赤潮生物的去除作用研究

有害赤潮对海洋生态环境和沿海经济危害巨大,如何安全、有效地治理有害赤潮非常重要。文章考察了几种常见芽孢杆菌对近海典型赤潮生物——东海原甲藻(Prorocentrum donghaiense)的去除作用,发现解淀粉芽孢杆菌(Bacillus amyloliquefaciens)对其去除作用最强。进一步考察解淀粉芽孢杆菌对不同赤潮生物的去除作用,发现该芽孢杆菌对米氏凯伦藻(Karenia mikimotoi)和赤潮异弯藻(Heterosigma akashiwo)的去除作用高于东海原甲藻。在此基础上,将解淀粉芽孢杆菌与高岭土复合,研究了微生物复合粘土对典型赤潮生物的去除效果。结果表明,解淀粉芽孢杆菌与高岭土复合后,能够有效促进该菌的生长,进而提升了其对赤潮生物的去除能力。文章对芽孢杆菌及其复合粘土去除赤潮生物的机制进行了分析和探讨,为进一步发展和优化改性粘土治理赤潮技术体系提供了重要参考。     

A study on relation between heterotrophic bacteria and shrimp disease

Ａ　ｓｔｕｄｙ　ｏｎ　ｒｅｌａｔｉｏｎ　ｂｅｔｗｅｅｎ　ｈｅｔｅｒｏｔｒｏｐｈｉｃ　ｂａｃｔｅｒｉａ　ａｎｄ　ｓｈｒｉｍｐ　ｄｉｓｅａｓｅ￥／／ＩＮＴＲＯＤＵＣＴＩＯＮＨｅｔｅｒｏｔｒｏｐｈｉｃｂａｃｔｅｒｉａ，ｗｈｉｃｈｏｂｔａｉｎｎｕｔｒｉｅｎｔ...     

南极菲尔德斯半岛土壤微生物在富集传代培养中对菲的响应

为揭示富集传代培养过程中南极多环芳烃降解菌群的演替规律,以菲为唯一碳源和能源对南极菲尔德斯半岛不同地理位置的6份土壤样品进行了富集和连续传代培养,并采用高通量测序技术分析了富集传代培养过程中菲降解菌群的群落结构及生物多样性。Alpha多样性分析结果表明,在富集培养阶段南极菲降解菌的物种丰富度显著高于传代培养阶段(P0.05),但传代培养阶段的代际物种丰富度无显著差异(P0.05)。Beta多样性分析结果表明富集培养阶段与传代培养阶段的群落结构具有显著性差异(P0.05),但传代培养阶段菲降解菌群趋于相对稳定,代际间群落结构无显著性差异(P0.05)。大量微生物在富集培养阶段丰度较高,但因不能适应实验室培养条件或不能利用菲而在传代培养阶段被淘汰。反之,假单胞菌属(Pseudomonas)、鞘氨醇菌属(Sphingobium)、贪噬菌属(Variovorax)、甲基娇养杆菌属(Methylotenera)和产碱杆菌属(Alcaligenes)等适应了该选择压力并在传代培养阶段形成了动态平衡的优势种群。此外,Pseudomonas、Sphingobium和Variovorax等在富集传代培养过程中具有显著性差异的优势种在维持群落的动态平衡中也发挥着重要的作用。该研究有助于加深人们对南极土壤环境中持久性有机污染物的降解潜力以及关键微生物动态变化的认识,为下一步充分挖掘利用南极环保功能微生物资源提供重要的依据。     

海洋污损细菌分离纯化及金属离子抑菌研究

为了研究海洋生物污损对船体和海洋工程的影响,作者将热喷涂镀钴基、镍基及含铜氧化钛基膜的船用厚钢板试样片放置在海水中,利用振荡培养箱模拟海洋环境,进行晃片培养。分离纯化镀膜钢片表面上的细菌,研究早期污损细菌的生长规律,通过革兰氏染色、生理生化实验对细菌进行鉴定,伯杰氏系统细菌学手册查证。采用纸片法测定抑菌圈,研究铜、钴、镍3种金属离子对污损细菌的影响,观察不同离子浓度的抑菌圈。研究结果表明:早期海洋污损细菌有G~+芽孢杆菌属(Bacillus)、G~+芽孢乳杆菌属(Sporolactobacillus)、G~+无芽孢杆菌(Bacilli)、葡萄球菌属(Staphylococcus)、G~+无芽孢丝状菌、假单胞菌属(Pseudomonas)6类;其中G~+细菌首先吸附; Cu~(2+)的最佳抑制质量浓度为30 g/L, Co~(2+)的最佳抑制质量浓度为80 g/L, Ni~(2+)的最佳抑制质量浓度为110 g/L。     

过硫酸氢钾复合物对对虾养殖底泥硝化作用、氨氧化微生物丰度和群落结构的影响

采用在模拟池塘中投放过硫酸氢钾复合物（KMPS）进行对比实验的方法,探究KMPS对养殖底质硝化作用的影响。通过对氨氮和亚硝态氮含量的检测,探究对不同时期氮素转化的影响,低频率高剂量投放组中的氨氮和亚硝态氮含量显著降低,而高频率低剂量组中氨氮和亚硝态氮的含量显著上升。高频率低剂量KMPS的投放使氨氧化古菌（AOA）和氨氧化细菌（AOB）丰度降低,而低频率高剂量KMPS的投放不会造成丰度降低,而且还表现出部分时期AOB丰度的上升。进一步对群落结构进行分析发现KMPS的投放使AOA群落中Nitrosopumilu属相对丰度降低,Nitrososphaera属的相对丰度上升,这种相对丰度的变化与KMPS的投放方式无关;但AOB群落受到KMPS投放方式的影响,低频率高剂量的KMPS投放下AOB群落优势属相对丰度显著提高。以上结果均说明低频率高剂量KMPS的投放起到了促进底质硝化作用的效果。同时,可为KMPS用于对虾养殖池塘底质改良开辟一个新的途径。     

从养殖贝类和大菱鲆分离的海洋发光弧菌的分类地位分析

海洋发光弧菌是海水养殖中重要的条件致病菌。从福建省晋江市附近水产养殖场花蛤、牡蛎软组织及波罗地海大菱鲆肠道内含物中分离得到16 株发光细菌, 采用ARDRA（amplified ribosomal DNArestriction analysis）、16S rDNA 序列测定、Biolog 碳源代谢分析及药敏试验等方法对...