改性粘土对虾夷扇贝(Mizuhopecten yessoensis)体内麻痹性贝毒的影响

目前,改性粘土技术已成为国内外普遍认可的有害赤潮治理方法,但在利用该技术治理产毒藻赤潮过程中,对底栖滤食性贝类体内毒素累积和排出的影响尚不清楚。通过室内实验,考察了聚合氯化铝改性粘土(MC I)絮凝典型产毒藻——太平洋亚历山大藻(Alexandrium pacificum)后,虾夷扇贝食用组织和消化腺组织中麻痹性贝毒(paralytic shellfish poisoning,PSP)含量、组分的变化情况。研究结果表明,在前期毒素累积阶段,虾夷扇贝暴露于A. pacificum 3 h后,未添加改性粘土的对照组中虾夷扇贝食用组织毒素水平由空白组的未检出迅速升高至13.24µg STXeq/kg,消化腺组织毒素水平由0.68µg STXeq/kg升高至42.97µg STXeq/kg,第4 d时对照组食用组织、消化腺组织毒素水平达到最高,分别为258.67和3 208.40µg STXeq/kg。添加0.2 g/L MC I的实验组中,3 h后水体中97%的A.pacificum藻细胞被絮凝沉降,此时虾夷扇贝食用组织和消化腺组织毒素水平远低于对照组,分别为6.33和18.39µg STXeq/kg,第2 d时达到98.92和574.54µg STXeq/kg,均显著低于对照组(P<0.05),随后呈下降趋势。在随后的毒素排出阶段,实验组虾夷扇贝食用组织和消化腺组织毒素水平显著低于对照组(P<0.05)。另外,在累积阶段,对照组和实验组虾夷扇贝食用组织和消化腺组织中PSP各组分相对含量差异不大,说明改性粘土未对虾夷扇贝体内的PSP转化造成明显影响。由实验结果可见,改性粘土可有效絮凝去除水体中的A.pacificum藻细胞,减少虾夷扇贝对其的滤食,从而降低了PSP在贝类体内的积累,该研究结果将为改性粘土治理底栖贝类养殖水体有毒赤潮提供科学依据。     

海产贝类车轮虫病及控制途径初探

报道山东沿海海产双壳贝类鳃丝上寄生的三种车轮虫：扇贝车轮虫、竹蛏车轮虫及蛤蜊车轮虫。同时对所涉种类的寄主专一性、感染途径、淡水杀灭效果及其危害性做了初步探讨。     

自然环境下虾夷扇贝(Patinopecten yessoensis)体内麻痹性贝毒的净化研究

为探究在自然环境中虾夷扇贝Patinopecten yessoensis体内麻痹性贝毒(PSTs)的净化过程,从黄海北部大窑湾海域采集了受PSTs污染的虾夷扇贝,将其转移至黄海的棋盘磨水域,该水域未曾有PSTs污染的报道。本研究采用高效液相色谱-荧光检测法(HPLC-FLD)检测30天净化过程中虾夷扇贝体内PSTs的组分和水平。研究结果发现,在第9天,虾夷扇贝软体中的PSTs的含量下降到一个相对较低的水平。而且,初期扇贝体内PSTs的净化率高于后期。扇贝各组织器官中PSTs含量分析发现,消化腺中的PSTs含量最高,这对于人类健康和扇贝养殖具有重要的意义。在本研究中还发现,虾夷扇贝的死亡率与PSTs的水平有一定的关系。     

几何形态学对8种扇贝的形态分类的应用

几何形态学(morphometrics)是基于笛卡尔地标点的统计分析方法,主要运用广义普鲁克分析(GPA)、薄板样条分析(TPS)、主成分分析(PCA)、典型变量分析(CVA)等方法,定量的对形态变化进行识别,结果更为客观。本研究运用几何形态学方法对8种常见扇贝(栉孔扇贝Azumapecten farreri、虾夷盘扇贝Mizuhopecten yessoensis、海湾扇贝Argopecten irradians、平濑掌扇贝Volachlamys hirasei、北海道扇贝Swiftopecten swiftii、荣套扇贝Gloripallium pallium、华贵类栉孔扇贝Mimachlamys nobilis、美丽环扇贝Annachlamys striatula)间形态关系进行研究,运用界标点和半界标点对扇贝的壳盘和壳耳进行数字化标点,利用PCA、CVA、TPS方法,获得8种扇贝的形态差异并进一步分析,建立系统发育树。PCA和CVA分析结果表明可以通过几何形态测量学分析方法将8种扇贝基于形态差异进行有效区分。此外,结合TPS的分析结果,发现壳耳和壳盘在扇贝中具有种间规律性差异。通过其形态分异进行表型聚类,从结果可知:美丽环扇贝、虾夷盘扇贝和海湾扇贝距离较近,壳型有共同的特征:扇贝壳盘较椭圆,壳耳相对较小;荣套扇贝和华贵类栉孔扇贝距离较近,壳型有共同的特征:扇贝壳盘更扁平,前耳相对较大;北海道扇贝、平濑掌扇贝和栉孔扇贝与其他种类没有交集。在聚类过程中,华贵类栉孔扇贝和荣套扇贝有交集,10个标本混淆,其他标本均成功聚类,正确率为95.5%。     

海水养殖鱼类外寄生车轮虫Ⅰ.车轮虫属

依据干银法和蛋白银染色法研究了山东沿海养殖鱼类的4种寄生车轮虫属纤毛虫: 直钩车轮虫、杜氏车轮虫、日本车轮虫及小袖车轮虫, 其中直钩车轮虫为我国的新记录.作者对采自不同宿主的直钩车轮虫种群的形态学特征进行了比较描述, 对所涉车轮虫的形态学、宿主、分布、流行情况及危害程度等做了综合报道.     

鲈鱼的鳃寄生车轮虫病及扫描电镜观察

报道鲈鱼的鳃寄生车轮虫病及病理组织的扫描电镜观察。病原鉴定为 :直钩车轮虫、杜氏车轮虫、日本车轮虫、劳牧小车轮虫及简单两分虫 5种车轮虫科纤毛虫 ,其中数量上占优势的为直钩车轮虫。扫描电镜观察结果表明 ,鳃寄生车轮虫可对宿主的上皮组织产生明显的机械性损伤并造成鳃丝粘液分泌增多及炎症反应。文中就海水养殖鱼类及野生鱼类由车轮虫寄生造成的危害及防治途径做了探讨。     

海水养殖鱼类外寄生车轮虫Ⅱ.拟车轮虫属、小车轮虫属及两分虫属

依据干银法和蛋白银法染色法研究了黄渤海沿岸经济鱼类寄生的车轮虫科内3属4种纤毛虫: 球核拟车轮虫、斜拟车轮虫、劳牧小车轮虫及简单两分虫.其中球核拟车轮虫和斜拟车轮虫为我国的新记录.文中对所涉车轮虫的形态学、宿主、分布、流行情况等做了报道.     

海水养殖鱼类外寄生车轮虫I.车轮虫属

依据干银法和蛋白银染色法研究了山东沿海养殖鱼类的 4种寄生车轮虫属纤毛虫 :直钩车轮虫、杜氏车轮虫、日本车轮虫及小袖车轮虫 ,其中直钩车轮虫为我国的新记录。作者对采自不同宿主的直钩车轮虫种群的形态学特征进行了比较描述 ,对所涉车轮虫的形态学、宿主、分布、流行情况及危害程度等做了综合报道     

大黄鱼抗菌肽hepcidin-like的抗菌和抗寄生虫活性分析

近年来，我国重要经济鱼类大黄鱼（Larimichthys crocea）遭受了严重的海水白点病的感染，造成了巨大的经济损失。海水白点病是由体外寄生虫刺激隐核虫（Cryptocaryon irritans）感染引起，不仅能引起被感染鱼类生理的破坏，还能够引发二次细菌感染。有报道指出一些抗菌肽（antimicrobial peptides）具有抗刺激隐核虫的活性。大黄鱼hepcidin-like（命名为Lc-HepL）是从刺激隐核虫感染大黄鱼后的比较转录组中挖掘到的一个差异表达基因。本研究中，在基因和蛋白水平上分析了该抗菌肽的生物活性。刺激隐核虫感染后，qRT-PCR检测到Lc-HepL在鳃、肌肉、肝脏、头肾、肠道和脾脏6种组织中显著上调，并且上调的时间点出现在幼虫感染阶段、滋养体脱落阶段和二次细菌感染阶段，结果显示Lc-HepL在抗刺激隐核虫和二次细菌感染的免疫反应中发挥重要作用。在体外成功诱导并纯化的重组Lc-HepL（rLc-HepL）对某些病原菌具有剂量和时间依赖性的强抗菌活性。本研究首次探究了rLc-HepL的抗刺激隐核虫活性，显微观察结果显示其能引起幼虫细胞膜破裂、内容物外泄，该结果首次提供了大黄鱼hepcidin-like具有强抗刺激隐核虫活性证据。研究数据表Lc-HepL很可能是大黄鱼抗刺激隐核虫感染的一种重要的先天免疫因子，具有应用到未来的药物中的潜力。     

海水养殖鱼类外寄生车轮虫II.拟车轮虫属、小车轮虫属及两分虫属

依据干银法和蛋白银法染色法研究了黄渤海沿岸经济鱼类寄生的车轮虫科内 3属 4种纤毛虫 :球核拟车轮虫、斜拟车轮虫、劳牧小车轮虫及简单两分虫。其中球核拟车轮虫和斜拟车轮虫为我国的新记录。文中对所涉车轮虫的形态学、宿主、分布、流行情况等做了报道。     

东印度洋5种缘毛类纤毛虫(钟虫属和聚缩虫)水平分布初次报道

腹壁纤毛虫是一种附着在浮游动物上的原生生物，尤其是附着在桡足类和甲壳类上的。而缘毛虫是可以以自由或寄生的形式在世界各沿海地区和港湾处被发现。本研究首次对东印度洋表层至200 m深海域的缘毛类纤毛虫以及其水平分布展开了研究。根据纤毛虫的形态、纤毛结构和茎的形态等的特征，在2014年4月10日至5月13日东印度洋航次的浮游生物拖网样品中鉴定出了五种富腹纤毛虫，海洋钟形虫Vorticella oceanica、交替聚缩虫Zoothamnium alternans、阿拉聚缩虫Zoothamnium alrasheidi、远洋聚缩虫Zoothamnium pelagicum和海洋聚缩虫Zoothamnium marinum。并通过光显微镜和扫描显微镜拍摄的照片，说明了其形态、尺寸和特征。研究结果表明，海洋钟形虫和远洋聚缩虫的分布与硅藻（如，密聚角毛藻Chaetoceros coarctatus）和桡足类（如，短角长腹剑水蚤Oithona brevicornis）等寄主有关，也包括甲藻（如，三角角藻Ceraitum tripose）周围的一些个体。这些附着纤毛虫的分布在东印度洋海域东南区以海洋钟形虫、远洋聚缩虫和海洋聚缩虫为主，而在孟加拉湾北部则有大量的阿拉聚缩虫。这种分布的变化可能是与寄主的分布有关，如桡足类（短角长腹剑水蚤）和硅藻（密聚角毛藻）的分布影响。     

热带西太平洋的小粟虫属的四种有孔虫(原生动物)的形态学研究以及Miliolinella obesa nov. spec. 新种描述

Foraminiferal oozes were sampled from the tropical West Pacific seamount and seabed of deep sea,by a remotely operated vehicle(ROV)in December 2014 and March 2016.Using standard morphological method,four Miliolinella species,including Miliolinella obesa,M.circularis,M.suborbicularis and M.subrotunda were described.Among the four species,M.obesa is a new species.It is a large member(about 500μm in length)of the genus.This species is characterized by having a very stout and transverse broadly circular outline,and its body width is greater than the body length.In addition,its chamber demarcations are obscure from the exterior appearance.Three chambers are unclearly visible on one side and two chambers are visible on the opposite.Finally,we provided very detailed taxonomic microphotographs and the ecological distribution information for each species.     

海草凋落叶的溶解有机物的释放及其生物地球化学意义

Dissolved organic matter(DOM) represents a significant source of nutrients that supports the microbial-based food web in seagrass ecosystems. However, there is little information on how the various fractions of DOM from seagrass leaves contributed to the coastal biogeochemical cycles. To address this gap, we carried out a 30-day laboratory chamber experiment on tropical seagrasses Thalassia hemprichii and Enhalus acoroides. After 30 days of incubation, on average 22% carbon(C), 70% nitrogen(N) and 38% phosphorus(P) of these two species of seagrass leaf litter was released. The average leached dissolved organic carbon(DOC), dissolved organic nitrogen(DON) and dissolved organic phosphorus(DOP) of these two species of seagrass leaf litter accounted for 55%, 95% and 65% of the total C, N and P lost, respectively. In the absence of microbes, about 75% of the total amount of DOC, monosaccharides(MCHO), DON and DOP were quickly released via leaching from both seagrass species in the first 9 days. Subsequently, little DOM was released during the remainder of the experiment. The leaching rates of DOC, DON and DOP were approximately 110, 40 and 0.70 μmol/(g·d). Leaching rates of DOM were attributed to the nonstructural carbohydrates and other labile organic matter within the seagrass leaf. Thalassia hemprichii leached more DOC, DOP and MCHO than E. acoroides. In contrast, E. acoroides leached higher concentrations of DON than T. hemprichii, with the overall leachate also having a higher DON: DOP ratio. These results indicate that there is an overall higher amount of DOM leachate from T. hemprichii than that of E. acoroides that is available to the seagrass ecosystem. According to the logarithmic model for DOM release and the in situ leaf litter production(the Xincun Bay, South China Sea), the seagrass leaf litter of these two seagrass species could release approximately 4×10~3 mol/d DOC, 1.4×10~3 mol/d DON and 25 mol/d DOP into the seawater. In addition to providing readily available nutrients for the microbial food web, the remaining particulate organic matter(POM)from the litter would also enter microbial remineralization processes. What is not remineralized from either DOM or POM fractions has potential to contribute to the permanent carbon stocks.     

Integration of the nuclease protection assay with sandwich hybridization (NPA-SH) for sensitive detection of Heterocapsa triquetra

Microalgae are photosynthetic microorganisms that function as primary producers in aquatic ecosystems. Some species of microalgae undergo rapid growth and cause harmful blooms in marine ecosystems. Heterocapsa triquetra is one of the most common bloom-forming species in estuarine and coastal waters worldwide. Although this species does not produce toxins, unlike some other Heterocapsa species, the high density of its blooms can cause significant ecological damage. We developed a H. triquetra species-specific nuclease protection assay sandwich hybridization (NPA-SH) probe that targets the large subunit of ribosomal RNA (LSU rRNA). We tested probe specificity and sensitivity with five other dinoflagellates that also cause red tides. Our assay detected H. triquetra at a concentration of 1.5×10⁴ cells/mL, more sensitive than required for a red-tide guidance warning by the Korea Ministry of Oceans and Fisheries in 2015 (3.0×10⁴ cells/mL). We also used the NPA-SH assay to monitor H. triquetra in the Tongyeong region of the southern sea area of Korea during 2014. This method could detect H. triquetra cells within 3 h. Our assay is useful for monitoring H. triquetra under field conditions.     

发现于我国渤海辽东湾的海洋甲藻新种-渤海异冒藻(Heterocapsa bohaiensis sp. nov.)

A small armed dinoflagellate bloomed in the aquaculture ponds off the coast of Liaodong Bay, Bohai Sea of China,resulting in heavy mortalities of the cultured prawns(Penaeus japonicus) and larvae of Chinese mitten handed crabs(Eriocheir sinensis). The bloom-forming species was successfully isolated, and cellular morphology of the specimen was consequently investigated through light, fluorescent and electron microscopy. The small((14.4±1.6) μm in length) ellipsoid cells show typical Heterocapsa thecal plate arrangement(Po, cp, 5′, 3 a, 7′′, 6 c,5 s, 5′′′, 2′′′′). The episome is evidently bigger than the hyposome. One to three spherical pyrenoids are located above or beside the large elongated nucleus. The body scale is characterized by a triangle basal plate with one central upright and nine peripheral spines. Above all, Heterocapsa bohaiensis could be distinguished from other Heterocapsa species by the combination of the cell size, morphology, cellular structure and body scale. Sequence analyses of both ITS and LSU regions reveal the significant genetic divergence between H. bohaiensis and other established species in this genus, further supporting novelty of this species. Noticeably, different sample treatment methods resulted in morphological variation of the apical pore complex(APC) of H. bohaiensis, which needs to be taken into account in future study.     

中国东南沿海小头虫科(环节动物:多毛类)一新种

中蚓虫是小头虫科较为常见的属，中国海区的样品过去常被鉴定为加洲中蚓虫Mediomastus californiensis Hartman。本研究中，来自东南沿海四个不同海区的118条中蚓虫样品被鉴定为一个新的种类，在此给出具体描述。这个新种具有如下特征：（1）第1至9刚节表皮双环型，刚毛束位于体节中间位置，而后的刚节多环型，刚毛束位于体节后半部；（2）后腹区具有疣足突起；（3）前4个胸刚节仅具双羽型翅毛状刚毛，其它体节仅具巾钩刚毛；（4）胸区巾钩刚毛的柄长且直，巾长为宽的3至5倍；（5）腹区巾钩刚毛不同于胸区，较短且具收缩部；（6）甲基绿染色模式独特。这个研究表明了中国海区的中蚓虫可能被误鉴定为加洲中蚓虫，而这个新种M.chinensis sp.nov.在东南沿海广为分布。     

利用扩增vvhA基因的环介导等温扩增技术快速，特异而敏感地检测创伤弧菌

创伤弧菌是一种可感染人类的河口病原菌。建立快速，特异而敏感的检测方法，有助于创伤弧菌感染的早期疾病诊断和及时治疗。本研究设计了针对vvhA基因的一系列引物（包括两对外部引物和两对内部引物），采用环介导等温扩增技术（LAMP）来检测创伤弧菌。结果显示，本方法的最适扩增温度是63℃，反应仅需35分钟。扩增产物不仅可以用含有DNA ladder的琼脂糖凝胶电泳检出，也可借助钙黄绿素直接肉眼观察。采用45株菌株检测该方法的特异性，其中所有创伤弧菌均被检出而其他菌株检测结果皆为阴性。本方法的敏感性是普通PCR扩增的100倍，同时利用该方法可以准确检测出所有的模拟样品、临床标本及环境样品。与其他已知方法比较，针对vvhA基因的介导等温扩增技术可以快速、简单、敏感及特异地鉴定创伤弧菌。     

基于稳定同位素分析推定秋冬季南极半岛南极大磷虾营养变化

南极大磷虾（Euphausia superba）为南大洋生态系统中的关键种，也是南极生态系统食物网中的重要枢纽。该种秋冬季转换期的营养信息对于理解其知之甚少的越冬机制非常重要。但关于此方面的少数研究在时空变化上仍存在着差异。为此，我们调查了南极半岛秋季（4-5月）和冬季（6月）磷虾成体δ¹³C和δ¹⁵N值的个体、月份及区域性差异。我们的目标旨在检验该期间磷虾的营养变化以及磷虾与其在南极海洋生态系统中的摄食环境之间的关系。结果如下：（1）磷虾δ¹³C值与体长之间无显著关系，但δ¹⁵N值与体长之间则存在显著相关性；（2）秋季磷虾δ¹³C值呈现增长趋势，但冬初季节并无显著变化，此期间δ¹⁵N值无显著不同；（3）布兰斯菲尔德与南设得兰群岛之间的δ¹⁵N平均值显著不同。我们的数据表明南极半岛秋至初冬转换期间磷虾成体营养呈现个体、季节性及区域性变化。     

西太平洋深海海水中乙醛降解菌多样性及群落结构研究

As a major aldehyde pollutant widely existing in industry and our daily life, acetaldehyde is more and more harmful to human health. As characteristic habitat niche, bacteria from deep sea environments are abundant and distinctive in heredity, physiology and ecological functions. Thus, the development of acetaldehyde-degrading bacteria from deep sea provides a new method to harness acetaldehyde pollutant. Firstly, in this study,acetaldehyde-degrading bacteria in the deep sea water of the West Pacific Ocean were enriched in situ and in the laboratory respectively, and then the diversity of uncultured bacteria was studied by using 16 S r RNA genes. Then acetaldehyde-degrading strains were isolated from two samples, including enrichment in situ and enrichment in laboratory samples of deep sea water from the West Pacific Ocean using acetaldehyde as the sole carbon source,and then the ability of acetaldehyde degradation was detected. Our results showed that the main uncultured bacteria of two samples with different enrichment approaches were similar, including Proteobacteria,Actinobacteria, Firmicutes, Cyanobacteria, but the structure of bacterial community were significant different.Four subgroups, α, γ, δ and ε, were found in Proteobacteria group. The γ-Proteobacteria was dominant(63.5%clones in laboratory enriched sample, 75% clones in situ enriched sample). The species belonged to γ-Proteobacteria and their proportion was nearly identical between the two enrichment samples, and Vibrio was the predominant genus(45% in laboratory enriched sample, 48.5% in situ enriched sample), followed by Halomonas(9% in situ enriched sample) and Streptococcus(6% in laboratory enriched sample). A total of 12 acetaldehyde-degrading strains were isolated from the two samples, which belonged to Vibrio, Halomonas,Pseudoalteromonas, Pseudomonas and Bacillus of γ-Proteobacteria. Strains ACH-L-5, ACH-L-8 and ACH-S-12,belonging to Vibrio and Halomonas, have strong ability of acetaldehyde degradation, which could tolerate 1.5 g/L acetaldehyde and degrade 350 mg/L acetaldehyde within 24 hours. Our results indicated that bacteria of γ-Proteobacteria may play an important role in carbon cycle of deep sea environments, especial the bacteria belonging to Vibrio and Halomonas and these strains was suggested for their potentials in government of aldehyde pollutants.     

分离自中国胶州湾的褐色马格里夫藻(Margalefidinium fulvescens)的形态学和系统进化特征

The dinoflagellate genus Margalefidinium has been split from Cochlodinium as a new genus recently and Margalefidinium fulvescens is one of the five Margalefidinium species. Margalefidinium fulvescens is toxic and has been reported from the coastal waters of USA, Canada, Mexico, China, Japan, Indonesia, Korea, Pakistan and Spain. Here we provide the morphological and phylogenetic characterization for an isolate of it from the Jiaozhou Bay, Qingdao, China. Our results showed that the vegetative cells were subspherical to ellipsoidal, 34–60 μm in length, and 19–41 μm in width. Both single cell forms and colonies in chains of 2, 4, or 8 cells were observed in cultures, but chain forms with 2 or 4 cells were observed more often in the field samples. The cingulum was rather deep, encircling the cell approximately twice, but the sulcus was rather narrow, surrounding the cell about one turn. The nucleus was spherical and located at the central epicone. The chloroplasts were granular, brownish, and scattered peripherally. An orange pigmented body also appeared in the epicone. The apical groove appeared vase-like as previously described. Under epi-fluorescence microscopy, a pumpkin-like structure was clearly observed, in which cells were embedded. Cells were observed to exit from the structure, which led us to a hypothesis that the structure may provide cells a shelter to avoid predation or to respond to other stresses. The phylogenetic analyses based on partial LSU rDNA sequences indicated that M. fulvescens from the Jiaozhou Bay was grouped with M. fulvescens populations from other origins and closely related to the clade of M. polykrikoides.Our morphological observations and phylogenetic analyses together confirmed the presence of M. fulvescens in China and our monitoring has also observed the species dominant in the dinoflagellate community of the Jiaozhou Bay in the early autumn of 2015, which alerted us to continually monitor this bloom-forming species in the region.