底栖硅藻大面积培养技术

底栖硅藻是从鲍上足分化匍匐幼体附着变态到长至壳长达3-5mm的稚鲍所必需的饵料。底栖硅藻质量的优劣，直接影响到鲍苗培育的成活率。因此培养足量的优质底栖硅藻是鲍人工育苗生产中最为重要的环节。本文重点围绕高密度、优质底栖硅藻规模化培养技术进行探讨，总结了一套底栖硅藻大面积培养技术，     

鲍采苗板上底栖硅藻种类变化的观察

底栖硅藻作为鲍幼体的前期饵料,其种类与质量对鲍人工育苗的成败起着至关重要的作用,其种类适宜且数量充足可以延长鲍幼体的附着变态率,延长在板时间,实现剥离大规格苗种,达到提高成活率的目的。据报道在底栖硅藻自然种群中,以小型的卵形藻(Cocconesp.)和舟形藻(Naviculasp.)的     

底栖硅藻中诱导鲍鱼幼体附着和变态的物质研究进展

底栖硅藻不但能提供促进鲍鱼幼体附着和变态的物质,还能作为鲍鱼幼体的饵料生物为其发育提供营养.底栖硅藻诱导鲍鱼幼体附着和变态的主要物质有胞外多糖、氨基酸、多不饱和脂肪酸等,对其营养功能和诱导附着变态作用等方面进行了综述,以期为鲍鱼育苗以及优质配合饲料的开发提供指导.     

明矾浓缩单胞藻饲喂刺参稚参的试验

刺参稚参营附着生活后，以附着基上的底栖硅藻和其他有机碎屑为饵料，这就需要在附着基上预先培养底栖硅藻。底栖硅藻的预先培养需在采苗前的15～20天进行，操作比较麻烦，并占用大量的空间，后期还需对底栖硅藻进行补充，生产上有较大的难度。在刺参人工育苗过程中，浮游单胞藻饵料需提前培养，往往出现生产过剩现象。特别是刺参幼体附着变态为稚参后，浮游单胞藻饵料的使用量明显减少，饵料室会剩余大量的单胞藻饵料，放弃不用则造成很大的浪费。     

九孔鲍育苗池附着基上底栖微藻群落及其多样性

2006年9月14至10月26日对福建省漳浦县下蔡九孔鲍(Haliotis diversicolor supertexta)育苗池进行定点采样。检测结果共鉴定底栖微藻24属62种,其中硅藻21属59种,绿藻、蓝藻和褐藻各1种。附着基上底栖微藻的优势种有12种,全部为硅藻,如盾卵形藻微小变种(Cocconeis scutellum var.parva)、小形舟形藻(Navicula parva)、咖啡双眉藻(Amphora coffeaeformis)、琴式菱形藻(Nitzschia panduriformis)等。优势种的优势度突出。常见种有25种,少见种有27种。优势种和常见种多是能够适应底栖环境的种类。附着基上底栖微藻种类及多样性指数育苗早期波动大,后期逐渐稳定,底栖微藻多样性指数平均为0.980～1.959,结果表明,底栖微藻群落结构的演变与鲍幼体摄食活动有很大的相关性。     

小型底栖硅藻的定向培养技术

皱纹盘鲍（Haliotis discus hannai）在育苗生产过程中,底栖硅藻的种类、质量,直接影响到鲍苗的附着率、成活率及生长速度,关系到整个育苗生产的成败。     

稚鲍饵料的初步试验

在鲍鱼人工育苗过程中，解决稚鲍的适宜饵料及满足供应的问题是充分发挥幼体的生长发育潜力，提高成活率的关键。如何解决稚鲍饵料问题，国内外曾有一些报道。如福建省东山鲍珠站的措施是：5mm以内的稚鲍，主要靠附着板上事先培养的底栖硅藻，其后以更换附着片进行饵料的补充。5mm以上幼鲍主要投喂海藻及人工配合饵料片。日本在5mm以下幼鲍期，主要也是靠底栖硅藻类及海带，     

底栖硅藻种类组成对皱纹盘鲍匍匐幼体和稚鲍生长、存活的影响

为验证由生产培养获得的不同质量的底栖硅藻培养皱纹盘鲍不同生长阶段幼体的效果,采用经不同培养措施获得的两类底栖硅藻群落(饵料A和饵料B)培养皱纹盘鲍前期匍訇幼体和稚鲍.试验结果表明,饵料A培养前期匍匐幼体的存活率[(43.47±13.53)%]显著高于饵料B[(6.94±5.17)%],但生长指标差异不显著.两类饵料培养的稚鲍存活率差异不显著,但饵料B的生长速度和特定生长率[(0.151±0.025)mm/d和(4.706±0.492)%/d)]显著高于饵料A[(0.103±0.022)mm/d和(3.625±0.374)%/d)].说明皱纹盘鲍幼体在不同生长阶段适应底栖硅藻的种类不同.匍匐幼体期对底栖硅藻质量要求较高,稚鲍期由于食量增加和食性转化,对饵料数量的要求表现得更为明显.     

2种底栖硅藻饲养杂色鲍幼体和稚贝的饵料效果

用羽纹藻（Pinnularia sp．）、简单双眉藻（Amprhoraexigua）、羽纹藻和简单双眉藻混合（混合组）作为饵料，以自然海区采集的硅藻作对照，研究其对杂色鲍（Haliotisdiversicolor）幼体和稚贝成活和生长的影响，确定人工分离的2种硅藻的饵料效果。结果表明，2种底栖硅藻在杂色鲍育苗中具有一定的饵料价值。饵料种类对不同规格杂色鲍的成活率和生长影响显著，并且2个观测指标的结果相同。幼体羽纹藻组、简单双眉藻、混合组3组间的成活率、生长无显著差异，但都明显比对照组低；稚贝简单双眉藻、混合组和对照组无显著差异，羽纹藻组显著比对照组低。     

不同浓度化学刺激物对多棘海盘车摄食行为及呼吸代谢的影响

实验室条件下,研究了3种贝类(太平洋牡蛎、紫贻贝、菲律宾蛤仔)不同浓度组织匀浆液及5种单一L型氨基酸(甘氨酸、谷氨酸、酪氨酸、天冬氨酸、赖氨酸)等化学刺激物对多棘海盘车(Asterias amurensis)摄食行为及呼吸代谢的影响。将多棘海盘车对刺激物的行为反应分为正趋向、负趋向和无反应3类,趋向反应由简单到复杂分为4种反应类型,通过测定耗氧率来表示呼吸代谢水平。结果显示,3种贝类组织匀浆液和5种氨基酸(浓度范围10 10~10 4mol/L,酪氨酸除外)均引起多棘海盘车的正趋向反应,且多棘海盘车产生反应的数量比例随刺激物浓度的升高而增加,反应类型也由简单逐渐趋于复杂。同种氨基酸在低浓度下引起多棘海盘车正趋向反应,但超过一定浓度(10 3~10 2mol/L)后会引起负趋向反应(实验浓度范围内谷氨酸组未出现)。不同种类刺激物对多棘海盘车摄食行为反应的影响不同,谷氨酸、甘氨酸以及菲律宾蛤仔组织匀浆液对多棘海盘车诱引效果最好。多棘海盘车的耗氧率随刺激物浓度的增加而逐渐升高。在贝类组织匀浆液中多棘海盘车的耗氧率均高于对照组;甘氨酸、谷氨酸、天冬氨酸各处理组中,只有在较高浓度下耗氧率才显著高于对照组,酪氨酸各组之间无显著差异。研究旨在为贝类底播增养殖和人工鱼礁区敌害生物的防除提供参考,同时为多棘海盘车觅食行为机制的研究提供数据支持。     

Induction of larval settlement and metamorphosis of the abalone, Haliotis discus hannai larvae using bromomethane and potassium chloride

Larvae of Haliotis discus hannai were induced to settle and metamorphose by exposure to seawater supplemented with bromomethane. The settlement and metamorphosis of veliger larvae were observed with bromomethane concentrations of 300, 600, 900, 1200, 1500 ppm on the benthic diatom plates. The highest settlement and metamorphosis rates of larvae were observed in bromomethane concentrations of 600 ppm.

Abalone larvae were induced to settle and metamorphose by exposure to seawater supplemented with potassium chloride. The settlement and metamorphosis of veliger larvae was observed in potassium chloride concentrations of 14, 19, 24 and 29 mM on the benthic diatom plates. The highest settlement and metamorphosis rates of larvae were observed in potassium chloride concentrations of 14–19 mM.     

僧帽牡蛎幼体附着和变态的诱导

采用外加人工化学物质检验对僧帽牡蛎眼点幼体的附着和变态诱导活性,结果表明去甲肾上腺素、野上腺素和L-多巴可有效诱导幼体不固着变态.在药物持续作用下,肾上腺素的最佳诱导浓度为10-5mol@L-1(变态率67.4%),去甲肾上腺素的最佳诱导浓度为5×10-5mol@L-1(变态率52.2%),L-多巴的最佳诱导浓度为10-5mol@L-1(变态率43.0%),而对照组变态率为0.幼体大多在药物作用12～24h之间完成变态.三种药物诱导的变态个体中有85%以上是不固着变态.比较多种附着基和EPI的变态诱导活性,显示同种贝壳的活性较肾上腺素还高.去甲肾上腺素诱导幼体变态效应对低盐海水有较强耐受能力,而对高盐海水适应性差.     

Settlement of larval top shell <Emphasis Type="Italic">Turbo (Batillus) cornutus</Emphasis> in response to several marine algae

ABSTRACT:     The settlement of larval top shell Turbo  ( Batillus )  cornutus in response to seven macroalgal and three benthic diatom species was examined in the laboratory. The percentages of larval settlement differed significantly among algal species, and were significantly higher on two red algal species ( Marginisporum crassissima , an articulated coralline alga, and Gelidium elegans , an agarophyte) than on the green algae, brown algae and benthic diatoms tested. Marginisporum crassissima induced larval settlement most strongly throughout the experiments. The percentage of larval settlement on a crustose coralline alga (unidentified species), which is known as a suitable substratum for abalone larvae, was much lower than on M. crassissima and G. elegans , which have morphologically complex growth forms . The percentage of settled individuals on ethanol-killed M. crassissima was much lower than on intact M. crassissima . The results indicate that physical structure is unlikely to be the main inducer of settlement; therefore, settlement of top shell larvae seems to be strongly induced by chemicals from the two red algal species. The specific distribution of juvenile top shell in the field may be at least partly determined by the strong induction of larval settlement by articulated coralline and gelidiacean algae.     

Effects of diet and temperature on post‐settlement growth and survival of the short‐spined sea urchin Strongylocentrotus intermedius

Effects of diet and temperature on post‐settlement growth and survival of a sea urchin Strongylocentrotus intermedius, which fed on one of six mono‐cultured benthic diatoms, gametophytes of a brown alga Undaria pinnatifida, a green alga Ulvella lens plus mixed benthic diatoms, or no food at 6, 9, 12 and 15°C, were examined. Digestion efficiency (DE), ingestion rate (IR), excretion rate (ER) and digested cell content index (DCCI) of each diatom species were measured. Growth rates largely differed among the dietary treatments from 7 days post‐settlement. Sea urchins fed on a diatom Cocconeis sublittoralis showed considerably higher growth (7.85‐34.67 μm day^?1) than those fed on the other diets and high survival rates (100%). DCCI was also highest in C. sublittoralis. These results suggest that the quantity of diatom cell contents digested is an important factor affecting growth and survival of the sea urchins. Growth rates of sea urchins were higher in higher water temperatures, and significant differences in growth rate were observed between 6‐9 and 12‐15°C. ERs of C. sublittoralis were generally higher at higher temperatures, but DEs did not differ among the temperature treatments. Variations in growth rate among different water temperature appear to be caused by different food intakes of the urchins.     

Free amino acids in metamorphosing bonefish (Albula sp.) leptocephali

Metamorphosing leptocephalous larvae of the bonefish (Albula sp.) were analyzed for total ninhydrinpositive substances (NPS) and free amino acids. Total NPS content showed little change during metamorphosis. The average NPS value (±S.E.) for 16 larvae was 1.8 (±0.1) mg×larva^–1, which represents approximately 4% and 8% of the total dry weight of early and advanced larvae, respectively. Taurine was the most abundant free amino acid in whole-larva extracts, accounting for 36% and 59% of the total by weight in early and advanced larvae, respectively. The essential amino acids, leucine, isoleucine, phenylalanine, histidine, valine, methionine, lysine and arginine, accounted for about half (47%) of the total in early larvae but were reduced to about 23% of the total in advanced larvae. All of the component essential amino acids decreased during metamorphosis, but the greatest effect was seen with the first five. The remaining non-essential amino acids comprised less than 20% of the total in early larvae and, although the overall value changed little during metamorphosis, certain components such as glycine and glutamic acid showed large increases whereas others such as tyrosine and serine were reduced. Increases in amino acid content after acid hydrolysis of whole-larva extracts indicated that trichloroacetic acid-soluble, low molecular weight peptides were present in both early and advanced leptocephali.     

赤水河流域秋季底栖硅藻群落结构特征及水质评价

为研究赤水河流域底栖硅藻群落特征及水质状况,2019年9月测定了赤水河流域36个代表性样点的水体理化指标,调查了底栖硅藻群落组成,并应用硅藻指数评价了赤水河流域的水质状况。结果表明：秋季赤水河流域上游的电导率和总磷、总氮浓度均高于下游;调查采集到底栖硅藻29属138种,其中扁圆卵形藻多孔变种(Cocconeis placentula var. euglypta)、变异直链藻（Melosira varians）、极细微曲壳藻（Achnanthes minutissima）和谷皮菱形藻（Nitzschia palea）等为秋季赤水河流域优势种;下游底栖硅藻群落稳定性高于中上游;CCA结果显示可溶活性磷（SRP）和总氮（TN）是影响赤水河流域底栖硅藻群落的主要环境因子。TDI和CEE硅藻指数评价结果表明,赤水河流域水质等级整体为“中等”,55.6%的样点水质等级为“中等”以上,8.3%的样点水质等级为“极差”。     

单糖组合对藤壶金星幼虫附着的影响研究

藤壶Amphibalanus(=Balanus)amphitrite是典型的海洋污损生物,藤壶金星幼虫受外源信号的指引,寻找到合适的附着地点才能进行附着和变态。针对前期发现的有一定诱导活性的4种单糖Man、Xyl、Glc和Fuc,该研究利用正交试验研究其对藤壶金星幼虫附着的影响。结果发现,单糖组合对藤壶金星幼虫附着具有显著性影响,Man(5.00×10^-8 mol/L)、Glc(1.25×10^-8 mol/L)、Xyl(1.25×10^-8 mol/L)和Fuc(1.90×10^-7 mol/L)为最优组合。组合中4种单糖的影响力由高到低依次是Man>Fuc>Glc>Xyl,且Man、Fuc和Glc的影响具有显著性,Xyl对幼虫附着影响不大。添加单糖除了影响金星幼虫的附着率以外,更重要的是能显著缩短幼虫附着的时间进程。     

Culture of benthic diatoms and effect of dissolved organic carbon on metamorphosis of the ‘white sea urchin Lytechinus pictus (Verrill)’

The effect of biofilms of the benthic diatom Navicula incerta and of the dissolved organic carbon (DOC) they produce was evaluated on the growth and survival of Lytechinus pictus postlarvae during metamorphosis. In each container, 300 postlarvae were exposed to biofilms beginning on days 1, 3 and 6 of metamorphosis, using three replicates and a blank containing only filtered water. The experiments were run for 15 and 18 days in 500‐mL containers with daily water changes. We measured 30 postlarvae from each container and counted all organisms on days 1, 3, 9 and 15; DOC was quantified on days 3, 6 and 9. A second experiment was conducted to evaluate whether the presence of diatoms or the DOC they produced improved postlarval development. Five assays were performed in triplicate: a positive control with biofilms, a negative control with filtered water and three treatments with DOC‐enriched water from a N. incerta culture supplied on days 1, 4 and 8. In both experiments, postlarvae attained the largest size and highest survival in the treatments with the biofilm. The results indicate that the increase in postlarval size and survival was due to the DOC produced by the diatoms and not by their presence alone.     

维生素B7和B12对细菌生物被膜形成及厚壳贻贝幼虫变态的影响

为探究B族维生素对海洋细菌生物被膜形成、海洋贝类幼虫变态所产生的作用,本研究首先使用维生素B7 (VB7)和B12 (VB12)直接刺激厚壳贻贝(Mytilus coruscus)幼虫,观察其对变态的直接诱导活性;然后通过添加VB7和VB12,与海假交替单胞菌(Pseudoalteromonas marina)共同形成生物被膜,分析B族维生素对生物被膜形成及其生物学特性的影响;同时检测生物被膜变化对厚壳贻贝幼虫变态发育的影响。研究结果显示,0.02 mmol/L浓度VB7和VB12可以直接诱导厚壳贻贝幼虫的变态,且效果最为显著(P<0.05);0.02 mmol/L浓度VB7和VB12处理后的海假交替单胞菌生物被膜对幼虫附着变态的诱导作用均显著提高(P<0.05);进一步通过细菌密度计数、膜厚度分析、可拉酸染色和定量等方法,揭示VB7和VB12处理后生物被膜细菌密度、膜厚度以及胞外多糖、蛋白和脂质均显著增加(P<0.05)。研究结果证实,VB7和VB12可能通过改变海洋细菌生物被膜的生物学特性,进而调控厚壳贻贝幼虫变态发育。本研究为探究厚壳贻贝幼虫附着变态的分子机制提供了新的理论依据和创新思路,同时为B族维生素在提高厚壳贻贝人工育苗技术、改善厚壳贻贝养殖产业问题和促进海洋牧场生态修复建设等方面的应用提供了理论基础。     

脂多糖对细菌生物被膜形成及厚壳贻贝幼虫变态的影响

为探究脂多糖(LPS)对海洋细菌生物被膜形成、海洋贝类幼虫变态所产生的作用,本研究使用不同浓度的LPS直接刺激厚壳贻贝幼虫,观察其对幼虫附着变态的直接作用;同时在海假交替单胞菌形成生物被膜的过程中添加不同浓度的LPS,通过分析生物被膜生物学特性的变化,及变化后的生物被膜对厚壳贻贝幼虫附着变态的影响。研究结果显示,3种浓度的LPS均可直接诱导厚壳贻贝幼虫的变态;10 mg/L浓度LPS处理后的生物被膜其细菌密度、膜厚明显降低(p<0.05),胞外产物中多糖(如可乐酸)、脂类显著增加(p<0.05),同时其对幼虫变态的诱导作用也显著提高(p<0.05),因此推测,在厚壳贻贝幼虫的附着变态过程中,LPS具有直接诱导作用;除此之外,脂多糖还可以通过调控生物被膜胞外物质,特别是可乐酸的生成,从而间接影响厚壳贻贝幼虫的附着变态。