塔玛亚历山大藻对两种单细胞藻生长的影响

本文研究了塔玛亚历山大藻(ATCI01品系)细胞再悬浮液、细胞破碎液、上清液、藻壳液、过滤液对亚心形扁藻悬浮性的影响,以及塔玛亚历山大藻与亚心形扁藻和小球藻之间的相互作用.结果发现,含有塔玛亚历山大藻成分的实验溶液均能在短时间内造成亚心形扁藻失去鞭毛形成胞囊而下沉,其中再悬浮液的影响最大.但亚心形扁藻受到的这种不利影响,一般在一周后即开始恢复.小球藻生长几乎不受塔玛亚历山大藻的影响,混合培养水体中的小球藻生长曲线与单种培养的几乎一致,其悬浮性也无明显改变.与亚心形扁藻进行混合培养的塔玛亚历山大藻(ATCI01品系和ATHK品系)均表现出培养前期增殖曲线与单种培养类似,但在一周后混合培养的塔玛亚历山大藻的增殖曲线开始下降,这一时间恰好与扁藻细胞密度开始增加的时间相一致.而与小球藻的混合培养的塔玛亚历山大藻在培养前期与单种培养一致,后期则种群数量有所下降.     

Effects of the toxic dinoflagellate Alexandrium tamarense on the greenshell mussel Perna canaliculus

Abstract

Oxygen consumption, grazing rate, and byssus production of the greenshell mussel Perna canaliculus were measured following exposure to bloom concentrations (10⁶ cells l^?1 ) of a toxic clone of the dinoflagellate AIexandrium tamarense (GT429), using anon‐toxic clone of the same species (PLY173) as control. Following feeding for 1 h on GT429, mussels (dry weight 54–127 mg) showed significantly increased oxygen uptake at 15°C (0.08 μl O₂ (mg dry tissue weight)^?1 ) compared with control mussels. However, following 24 h recovery, oxygen uptake was similar to initial values in all experimental conditions. The grazing rate of P. canaliculus fed upon toxic clone GT429 was 4.58 ± 0.76 cells mg^?1 h^?1 compared with 1.99 ± 0.47 cells mg^?1 h^?1 for PLY173. On exposure to A. tamarense, mussels maintained normal opening behaviour and there was no change in the byssus production over 24 h. It was concluded that P. canaliculus showed no dramatic physiological effects following short‐term feeding on the toxic strain of A. tamarense. During 2 weeks’ exposure to twice daily bloom concentrations of GT429, there was no mussel mortality. Toxicity in the tissues was 1295 μg STX equivalent per 100 g tissue, levels at which these mussels would be unsuitable for human or animal consumption.     

塔玛亚历山大藻对卤虫干重及能值的影响

研究了有毒赤潮藻塔玛亚历山大藻Alexandrium tamarense对甲壳动物卤虫的干重及能值的影响.实验表明,经过18h的染毒实验,7 500尾卤虫Artemia parthenogenetica在水体中有塔玛亚历山大藻细胞或过滤液(细胞密度均为200cells·ml-1)存在的情况下,其总干重及总能值的增加明显受到抑制.卤虫总干重,对照组增加0.010 4g,而滤液组和藻细胞组均增加了0.002 2g,仅为对照组的21.1%;卤虫总能值,对照组增加0.143 5kJ,而滤液组和藻细胞组则分别增加0.018 0kJ和0.056 8kJ,分别为对照组的39.5%和12.5%.经SAS软件分析,塔玛亚历山大藻细胞对卤虫总干重的影响与对照组比较有显著性差异(p=0.037 0),而对其总能值增加则无显著差异(p=0.1251);而塔玛亚历山大藻过滤液对卤虫的总干重、总能值的增加与对照组都有显著性差异(p分别为0.021 5和0.013 5).另外,无论是藻细胞或是过滤液对卤虫的比能值均无影响.有毒塔玛亚历山大藻使卤虫总干重减少可影响其他相关海洋动物的食物供给,而对卤虫能值的影响则表明毒藻可改变食物链中能量的传递效率,从而影响处于较高营养层的生物.研究还显示该藻的不利影响可出现在赤潮发生期及消退期,这为全面了解赤潮危害提供新的视角.     

Flushing rate and salinity may control the blooms of the toxic dinoflagellate Alexandrium tamarense in a river/estuary in Osaka Bay, Japan

The first observation of a red tide of the toxic dinoflagellate Alexandrium tamarense in the Yodo River estuary, Osaka Bay, Japan was in the spring of 2007. To clarify which environmental factors controlled the abundance of A. tamarense, field surveys were conducted in 2008 and 2009. In 2008, the increase of A. tamarense occurred from early to mid-March and temperatures ranged from 9 to 13 °C. In contrast, in 2009, the increase was 1 month later, from early to mid-April and therefore temperatures were higher. In both years, A. tamarense was most abundant when salinities were relatively high (15–25), river discharge was low, and the water column was stable. There were no cells during periods of low salinity (<10), high discharge. In 2008, possibly silicate and phosphate were limiting during the simultaneous diatom and A. tamarense bloom in early March with silicate restricting the further growth of diatoms. In this estuarine basin, freshwater discharge from the weir increased the flow rate at stations downstream of the weir and deterred bloom formation by A. tamarense. Conversely, the lack of discharge increased the salinity and stability of the water column and decreased in the flow rate in the river/estuary. This process promoted the initiation of the bloom of A. tamarense and the subsequent accumulation of paralytic shellfish toxin in the brackish water clam Corbicula japonica. It is recommended that a constant discharge from the weir is maintained in order to prevent blooms of A. tamarense and subsequent paralytic shellfish poisoning (PSP) incidents in this area.     

塔玛亚历山大藻生成二甲基硫和二甲基硫丙酸的实验研究

主要研究在封闭培养条件下塔玛亚历山大藻（Alexandrium tamarense)生长周期内藻体细胞的二甲基硫丙酸（DMSP（）含量以及释放至水体的二甲基硫（DMS）含量，结果表明：（1）塔玛亚历山大藻藻体细胞DMSP含量变化与该藻细胞数量动态变化趋势相一致，在生长周期的第7天最高值；（2）藻体细胞的DMSP含量以及释放至水体的DMS含量均与藻体细胞数量有显著相关；（3）单位细胞DMSP生成量的变化与DMS释放量变化呈现相反的趋势，在DMS释放量最高时，单位细胞DMSP生成量最低。     

海洋有毒甲藻Coolia monotis的研究概况

甲藻(Pyrrophyta)又称双鞭毛藻(Dinoflagellates),是海洋浮游植物的重要种群之一,其种类繁多,全世界已记录的约有130个属,1200个代表种。随着赤潮的频繁出现及赤潮毒素的强效活性的发现,有毒甲藻的危害及医药作用日益引起人们的重视。赤潮毒素是海洋生物活性物质中研究、开发进展最为迅速的领域之一。     

磷酸盐不同补充方式对塔玛亚历山大藻生长和产毒影响

实验室条件下探讨经过磷酸盐限制后,2种不同补给方式对塔玛亚历山大藻生长和产毒的影响.结果表明,经磷限制后,在一次性添加和脉冲式添加2种磷酸盐补充方式下,塔玛亚力山大藻的最大比生长速率分别为0.71 d-1和0.63 d-1,明显高于在f/2培养液条件下的0.40d-1;同时,一次性添加组所达到的最大生物量明显高于脉冲式添加组(P＜0.05),表明在寡磷酸盐海区,一旦磷酸盐得到丰富补充后,浮游植物会暴发性增长,极易引发赤潮.但是,2组在光能转化效率Fv/Fm值和毒素变化方面都没有表现出显著差异(P＞0.05).前6d受到磷酸盐限制的塔玛亚历山大藻,其光能转换效率值Fv/Fm从0.62降至0.40,单位细胞毒素含量却是在f/2培养液条件下的6倍;而添加磷酸盐后,Fv/Fm值在3d内迅速升至0.60,细胞数量呈现快速增长态势,单位细胞毒素含量却开始降低.至培养结束时,Fv/Fm值缓慢降低至0.40左右,单位细胞毒素含量分别为0.73和0.78 pgSTXeq/个,与f/2培养液条件下的0.76 pgSTXeq/个相当.HPLC测定结果表明,塔玛亚历山大藻南海株毒素含量主要是由GTX1,GTX4,neoSTX和C2四类组分构成,其和占总毒素含量的98％以上.     

厦门湾沉积物中有害有毒甲藻休眠孢囊及塔玛亚历山大藻的潜在危害

本工作通过分析厦门湾表层沉积物中的甲藻休眠孢囊，发现了1种有毒塔玛亚历山大藻(Alexandrium tamarense)休眠孢囊，及两种有害甲藻休眠孢囊：锥状斯氏藻(Scrippsiella trochoidea)、具刺膝沟藻(Gonyaulax spinifera)．对塔玛亚历山大藻进行了培养。并初步研究了该藻的毒性及贝类对其毒素的积累．研究结果表明，厦门海域沉积物中的塔玛亚历山犬藻是有毒的．它的较适宜生长环境条件是：水温20～25℃．盐度为25～30，光照度为3000～50001x．塔玛亚历山大藻在每年的4月26日至6月10日、10月11日至11月20日可能会形成有害藻华，危害厦门海域的海洋生物并可能威胁到人类的健康和生命安全．     

Study on impact of dinoflagellate Alexandrium tamarense on life activities of marine bivalves

The effects of a PSP producing dinoflagellate Alexandrium tamarense on marine bivalvesat their several important life stages: egg, D- shape larva, eyespot larva, juvenile and adult, were studied. The results show that the hatching, survival, activity, filtration and growth were adversely affected by the alga and the impact was significantly increased with the increase of algal density. The inhibitory effect on egg hatching was most significant, which the hatching rate was only 30% of the control when exposed to the alga at 100 cell/cm3 after 36 h. Further experiments show that the algal culture, re-suspended cells and cell fragments had the inhibitory effect, while no such effect was from the cell-free medium, cell contents and standard STX. The results indicate that the alga could produce unknown toxins, rather than PSP, associated with the cell surface.     

Effects of different nitrate and phosphate concentrations on the growth and toxin production of an Alexandrium tamarense strain collected from Drake Passage

Nitrate (N) and phosphate (P) are believed to be two of the most important nutrients for the growth and toxin production for Alexandrium species. The study of the growth and toxicity characteristics of the Alexandrium spp. under the change of N and P can help us to understand the dynamics of algal bloom and toxification events in natural environments. A strain of Alexandrium tamarense (designated as Kci) was successfully isolated from the Drake Passage in 2001 and the clonal culture has been kept in our laboratory (Ho et al., 2003, 2012). In order to extend our understanding on the growth physiology and toxicity of this A. tamarense strain, growth and cellular toxin content were examined in unialgal batch cultures under different concentrations of N and P. The effects of variable N, P concentrations on growth, cellular toxicity (fg STXeq. cell⁻¹), and toxin composition (% molar) were determined in both exponential and stationary growth phases. The toxin profile, determined by high-performance liquid chromatography with fluorescence detection (HPLC-FD), was found to be remained relatively stable and was consistently dominated by the N-sulfocarbamoyl C-toxins (>90%) under different conditions and growth phases. There were also trace amounts of other carbamate gonyautoxins consistently expressed. The cellular toxicity varied under different N and P concentrations, as well as different growth stages. A positive correlation was observed between cellular toxicity and N concentrations, but the toxicity was enhanced when P was depleted. Both cell densities and growth rate of the cells were severely suppressed under N- or P-depletion. However, the biovolume of the cells tended to be larger at N- or P-depleted cultures. Results from the present study provide valuable insight for the ecophysiology of Alexandrium species in the coastal ecosystem of Drake Passage.     

有毒甲藻—塔玛亚历山大藻在厦门地区虾塘引起赤潮

材料于１９９４年７月采自厦门及邻近地区对虾养殖池。塔玛亚历山大藻藻体细胞形态准圆形，长略大于宽。细胞和１６－３１μｍ，宽１３－３０μｍ。甲板排列公式为Ａｐ，４＇，６″，６ｃ，８ｓ，其孢囊长椭圆形，长径２２－２６μｍ，短径１８－２２μｍ，表面光滑。采样期间虾池表层水温２８－３１℃，盐度１５－３１。     

营养盐浓度和初始细胞密度对东海原甲藻与塔玛亚历山大藻种间竞争的影响

以我国东海2种重要赤潮原因种--东海原甲藻(Prorocentrum donghaiense)和塔玛亚历山大藻(Alexandrium tamarense)为研究对象,研究了高、低磷酸盐浓度以及初始细胞密度对这2物种种群增长与种间竞争的影响.结果表明,不同实验条件下2物种有不同的竞争结果,营养盐限制环境中主要表现为资源利用性竞争,东海原甲藻为竞争优势种;丰富营养盐环境中,种间竞争具有明显的密度依赖性,根据初始细胞密度的不同,2竞争物种既可以相互排斥,也可能共存.该结果与东海赤潮特征即2物种既可以分别形成单相型赤潮,又可以共同形成双相型赤潮相一致.     

微量元素Fe、Mn、Co对有毒甲藻生长和产毒的影响

实验通过培养基筛选实验和在此基础上的微量元素Fe、Mn、Co正交实验对海南三亚热带珊瑚礁海域有毒甲藻Coolia monotis的生长和产毒影响进行了初步研究,以期获得充足的原材料进一步开展毒素化合物的研究.实验结果表明,在Erd、f/2、GP和K 4种培养配方中,K系列培养液更适合于C monotis的培养.在以2K为基础培养配方的微量元素正交实验中,Fe、Mn和Co 3种微量元素的浓度配比为:Fe:2×10<'-5>mol·L<'-1>,Co:1×10<'-6>mol·L<'-1>,Mn:5×10<'-7>mol·L<'-1>时能获得最大的平均生长速率;而Fe:5×10<'-6>mol·L<'-1>,Co:1×10<'-7>mol·L<'-1>,Mn:2×10<'-6>mol·L<'-1>的条件下C monotis的毒性最强.     

东海原甲藻与塔玛亚历山大藻种群增长过程与种间竞争研究

针对我国东海2种重要赤潮肇事生物东海原甲藻(Prorocentrum donghaiense)和塔玛亚历山大藻(Alexan-drium tamarense)进行了种群增长过程和种间竞争的实验研究。结果表明,单种培养时,不同氮磷比对东海原甲藻和亚历山大藻种群增长过程没有明显的影响,但是会影响其最终的细胞密度;在混合培养时,营养盐组成比例不同对东海原甲藻和塔玛亚历山大藻的种间竞争过程有影响,但不会影响其最后竞争结果。竞争物种的细胞起始密度对种间竞争结果有明显的影响,P.donghaiense和A.tamarense的接种细胞密度比为2 000∶100和2 000∶200时,东海原甲藻占优势;当接种细胞密度比为4 000∶400时,塔玛亚历山大藻占优势。     

温、盐度及营养盐对不同藻株塔玛亚历山大藻生长的影响

利用2001年11月至2003年2月从长江口、闽江口和北部湾水域表层沉积物中分离的休眠孢囊,萌发获得三株塔玛亚历山大藻,并研究了温度、盐度、营养盐对它们生长的影响。实验结果表明,来源于三个不同海区的藻株在水温32℃的条件下均无法存活,温度对长江口藻株(ATDH01)及广西北部湾藻株(ATGX02)的生长有极显著影响(P<0.01),而对闽江口藻株(ATMJ02)的生长无显著影响(P>0.05)。不同海区的藻株,其生长特性有明显差异,ATDH01藻株最适生长温度为24℃,最适生长盐度为30,且在盐度15和35仍有较高的生长率;而ATMJ02藻株及ATGX02藻株最适生长温度分别为24℃和20℃,最适生长盐度范围为20～30,盐度超出此范围生长率明显降低。ATDH01藻株的生长繁殖能力最强。营养盐对三个不同海区藻株生长的影响基本一致,在低P(1.2μmol/L)组中,三株藻的指数生长期较短,细胞密度明显的低于f/2组;而在低N(44μmol/L)组中,虽然在培养初期,三株藻的细胞生长明显快于f/2组,但在生长后期由于氮的限制,最终细胞密度仍低于f/2组。     

翡翠贻贝对塔玛亚历山大藻的摄食及毒素积累的初步研究

塔玛亚历山大藻是一种可产生PSP毒素的海洋微藻，通过室内摄食实验，初步研究了厦门港水产养殖动物翡翠贻贝对塔玛亚历山大藻的滤食率与光照度、温度、藻液浓度等因素的关系，并对比了翡翠贻贝对其它饵料微藻的摄食压力．同时对比了不同细胞密度的塔玛亚历山大藻浓度对翡翠贻贝毒素累积的影响．     

两种海洋微藻的浓度和配比对太平洋纺锤水蚤生长的影响

本研究以变态存活率、产卵量、成体头胸部长和体长为指标,研究了球等鞭金藻(Isochrysis galbana)和海洋原甲藻(Prorocentrum micans)不同浓度和配比对太平洋纺锤水蚤(Acartia pacifica)生长发育和产卵量的影响,以期探索培养太平洋纺锤水蚤最适合的饵料配方.实验结果表明:在饵料浓度(碳含量,下同)为1.70μg/cm3,配比为2∶1时,其变态存活率最高,可达到92.62%;在饵料浓度为4.25μg/cm3,配比为1∶2时,产卵量最高,雌体平均日产卵量可达47.1±2.7个;在饵料浓度为1.70μg/cm3,配比为2∶1时,成体头胸部长和体长均最长,分别为932.9、1 252.3μm.太平洋纺锤水蚤生长和发育的最优饵料组合为球等鞭金藻与海洋原甲藻浓度为1.70μg/cm3,配比为2∶1.     

长江口3 种贝类碳、氮收支的研究

以春季长江口缢蛏(Sinonovacula constricta)、河蚬(Corbicula fluminea)和光滑河兰蛤(Potamocorbula laevis)为研究对象,研究了此3种滤食性贝类的摄食生理参数,并根据能量平衡原理估算了3种双壳贝类的碳、氮收支情况。结果表明,(1)3种双壳贝类从滤食藻类中摄取的有机碳源主要通过呼吸代谢消耗、以粪便的形式排出,少部分随排泄代谢产物流出,余下的碳主要用作贝类自身的生长。缢蛏的碳收支方程式为100.00C(摄食碳)=20.17F(粪便碳)+50.05R(呼吸碳)+9.86U(排泄碳)+19.92P(生长碳),河蚬的碳收支方程式为100.00C=31.29F+37.40R+5.05U+26.26P,光滑河兰蛤的碳收支方程式为100.00C=44.13F+33.08R+11.05U+11.74P。(2)由于贝类在呼吸代谢中没有氮排放,故3种贝类的生长氮占总摄食氮的比例较碳大。因此,缢蛏氮收支方程式为100.00C(摄食氮)=28.22F(粪便氮)+49.38U(排泄氮)+22.40G(生长氮),河蚬的氮收支方程式为100.00C=45.05F+23.99U+30.96G,光滑河兰蛤的氮收支方程式为100.00C=46.97F+32.95U+20.08G。     

不同起始浓度对塔玛亚历山大藻和赤潮异弯藻种群竞争的影响

初步分析了起始浓度对塔玛亚历山大藻 (AlexandriumtamarenseBalech)和赤潮异弯藻 (HeterosigmaakashiwoHa da)种群增长的影响。在单种培养的情况下 ,A .tamarense与H .akashiwo的生长曲线可用逻辑斯谛增长模型拟合。不同起始浓度 ( 0 .2× 10 4,0 .5× 10 4,0 .8× 10 4cell/mL)对A .tamarense环境容纳量 (K)有明显的影响 ,随起始浓度的增加K有减小的趋势。H .akashiwo种群瞬时增长率 (r)随起始浓度增加而增加。按不同接种比例进行混合培养 ,不同起始浓度对 2种藻竞争的结果具有明显影响。当接种比例为A .tamarense∶H .akashiwo(A∶H) =1∶4时 ,H .akashiwo在竞争中占优势 ,而当A∶H =1∶1及A∶H =4∶1时 ,A .tamarense在竞争中占优势。竞争结果不仅受营养盐限制 ,克生物质对竞争结果亦具有重要影响。