大窑湾养殖区赤潮甲藻孢囊种类组成及分布的研究

2006—2007年对大窑湾养殖区的甲藻孢囊进行了研究,结果检出10种甲藻孢囊,分属于4大类9种甲藻孢囊,其中原多甲藻类孢囊2种,膝沟藻类孢囊5种,钙质类孢囊1种,裸甲藻类孢囊1种,一种艉杆藻(Ensiculifera Balech)和一种易碎藻(Fragilidium mexicanum)是中国沿海新记录的两个种类,此外还有一种无法鉴定到种的一种原多甲藻孢囊。膝沟藻类孢囊的种类数占绝对优势,占50%,其次是原多甲藻类孢囊占30%,钙质类孢囊和裸甲藻类孢囊各占10%。10种孢囊中有3种异养型孢囊和7种自养型孢囊。并发现两种产麻痹性贝毒(PSP)的孢囊—塔玛亚历山大藻/链状亚历山大藻复合体(Alexandrium tamarense/A.catenella)和链状裸甲藻(Gymnodinium catenatum)。孢囊密度的季节分布呈现明显的双峰型,在2007年1月份和2007年的8月份出现了两个高峰。2007年的1月份达到了调查全年的孢囊密度最高值,达到了1.09×103 cysts/g湿重,2007年的8月份出现的第二次高峰期孢囊密度达到了0.86×103 cysts/g湿重。孢囊密度最低值出现在2007年的10月,只有280 cysts/g湿重。全年孢囊平均密度为695.5 cysts/g湿重。甲藻孢囊的多样性指数除了2007年的9月和10月以外,相差不大,变化范围为0.17—0.55,全年的平均值为0.46。     

中国黄藻纲植物新记录属和种

报道了采自中国山西、黑龙江、上海、浙江等地的中国黄藻纲植物新记录14种,隶属于7个属,其中椭球藻(属Ellipsoidion)、拟杆藻属(Bumilleriopsis)、异毛藻属(Heterotrichella)和异丝藻属(Heterothtix)4个属在中国是首次报道。     

锥状施克里普藻孢囊的多种形态特征

休眠孢囊是某些甲藻生活史的重要阶段,它被认为是甲藻赤潮爆发的种源。通过观察我国沿海海域采集的锥状施克里普藻（Scrippsiella trochoidea（Stein）Loeblich）休眠孢囊的类型以及形态特征,并将实验条件下形成的休眠孢囊与海区沉积物中休眠孢囊的形态特征进行比较,结果表明,锥状施克里普藻细胞在20°C、12小时光照/12小时黑暗、f/2培养基和f/2-N培养基中分别培养至第14天和第8天,可以观察到休眠孢囊的产生。培养30天后,培养液中有大量休眠孢囊,将藻液避光保存90天后,观察到培养基中有4种类型的休眠孢囊产生：（1）球形到椭球形（长25-39μm）,钙质外壁,稀疏或密被钙质附属物（长1-4μm）;（2）球形（直径为29-35μm）,钙质外壁,附属物非钙质（长0.5-1.5μm）;（3）球形到椭球形（长29-35μm）,钙质,表面光滑,无附属物;（4）球形（直径24-35μm）,有机外壁,无附属物。第1种类型的孢囊最为常见,分别占到2种培养条件下所得全部孢囊的77.70%和95.18%。从沉积物中所得的孢囊多属于第1种类型。分析了实验条件下形成不同类型休眠孢囊的原因和比例,同时探讨了海区沉积物中形成多种孢囊形态特征的原因,为锥状施克里普藻孢囊的生态研究提供了依据。     

扁裸藻属和鳞孔藻属的新分类群

细胞较小,长25～28m,宽7～13m,厚4～5m,正面观呈短杆形或矩圆形,前端平截,中央略凹人,两侧略弯或近于平行,后端近圆形或平截,具一乳头状小尾突,侧     

广东省淡水红藻弯枝藻属一种（Compsopogon sp．）的形态描述及分类地位探讨

2009年8月在广东省龙川县定南水河采集到一份弯枝藻属的标本,其形态特征介于弯枝藻（Compsopogon caeruleus）和疏枝弯枝藻（Csparsus）之间。植物体株高10～18cm,主轴直径180～270μm,微收缢;中轴细胞近球形和扁球形,横壁收缢明显,直径150-200μm,高100～110μm;皮层细胞1层,中轴细胞缢缩处为2—3层;多角形或卵形,长25~35μm,直径20-25μm;分枝直出至侧生,某些基部细胞特化为假根;分枝顶端渐尖、急尖或二叉状;单孢子球形,直径10—14μm。该标本可能为弯枝藻和疏枝弯枝藻在演化过程中的一种过渡类型。     

云南溪菜属一新种

植物体大形,草绿色,柄极短小,以盘状固着器着生,形状多样,有披针形、窄或宽椭圆形、倒卵形,有的具较深的纵裂,长3～14cm,宽0.7～8.1cm,边缘不规则浅波状并波状皱褶,顶端钝圆或广圆形,有的具短的尖顶,基部先为钝圆形,后扩展为心形;细胞为近圆形或长圆形,长6.5～11(～13)μm,宽5～9μm,通常每4个为一组,每4组排列为一区,     

两种胶毛藻科植物的培养研究

对胶毛藻科植物优美胶毛藻[Chaetophora elegans(Roth)Agardh]和羽枝竹枝藻[Draparnaldia plumosa(Vauch) Agardh]进行了室内培养,对它们自游动孢子萌发到成熟丝状体形成等生活史各阶段都进行了详细观察,并做了活体显微镜照相记录。结果显示:(1)优美胶毛藻的游动孢子较小,直径为9~11μm,顶端具4条等长鞭毛,自母细胞释放后,游动孢子固定在基质上萌发,萌发类型为直立式,顶端细胞发育为直立系统,其直立系统为二叉状分枝,无明显主枝,侧枝发达,分枝顶端细胞渐尖或毛状,各级分枝细胞大小基本相同,圆柱形,宽约为6~8μm,长约为10~12μm。基细胞发育为匍匐系统,首先产生匍匐枝,由它产生假根。随生长,直立系统下部的细胞产生次生假根,成熟的优美胶毛藻外观近球形,与野生胶毛藻的形态相似。(2)羽枝竹枝藻游动孢子产生于母细胞,游动孢子体积较大,直径约为30μm左右,顶端具4条等长鞭毛。游动孢子从母细胞内释放后,固定在基质上萌发,萌发类型亦为直立式,顶端细胞发育为直立系统,其直立系统为分枝丝状体,主枝粗壮,细胞圆柱形,长约为50~60μm,宽约为48~50μm,内有多个蛋白核;侧枝细胞为长圆形,长约为40~50μm,宽约为17~19μm,分枝顶端的细胞尖细或为毛状。基细胞发育为假根和匍匐枝。当藻体成熟时,直立系统下部的细胞产生次生假根。     

采自青海湖的刚毛藻属一新种——青海刚毛藻

对青海湖的丝状藻类进行了样品采集,分析引起水华的藻类种类和组成。基于SSU-LSU的联合进化分析和基于ITS的系统进化分析显示:形成青海湖水华的丝状藻为一种刚毛藻,该种与史氏刚毛藻Cladophora stimpsonii和散束刚毛藻Cladophora vagabunda具有较近的亲缘关系,属于海洋刚毛藻和淡水刚毛藻的中间过渡种类。和邻近种类的区别特征如下:该种生长于咸水生境,植物体纤细柔软,呈绿色或浅绿色,分枝旺盛,具明显的顶端生长和居间生长。细胞直径较小。主轴细胞直径30.0—90.0μm,细胞长宽比2.4—8.0。分枝细胞直径25.0—70.0μm,长宽比4.0—12.3。顶端细胞直径25.0—50.0μm,长宽比3.6—12.0。基于形态学特征和分子系统发育分析,将其定为一新种,即青海刚毛藻Cladophora qinghaiensis sp. nov.。     

锥状施克里普藻孢囊的多种形态特征

休眠孢囊是某些甲藻生活史的重要阶段, 它被认为是甲藻赤潮爆发的种源。通过观察我国沿海海域采集的锥状施克里普藻(Scrippsiella trochoidea (Stein) Loebl ich)休眠孢囊的类型以及形态特征, 并将实验条件下形成的休眠孢囊与海区沉积物中休眠孢囊的形态特征进行比较, 结果表明, 锥状施克里普藻细胞在20°C、12小时光照/12小时黑暗、f/2培养基和f/2-N培养基中分别培养至第14天和第8天, 可以观察到休眠孢囊的产生。培养30天后, 培养液中有大量休眠孢囊, 将藻液避光保存90天后, 观察到培养基中有4种类型的休眠孢囊产生: (1)球形到椭球形(长25-39 μm), 钙质外壁, 稀疏或密被钙质附属物(长1-4 μm); (2)球形(直径为29-35 μm), 钙质外壁, 附属物非钙质(长0.5-1.5 μm); (3) 球形到椭球形(长29-35 μm), 钙质, 表面光滑, 无附属物; (4) 球形(直径24-35 μm ), 有机外壁, 无附属物。第1种类型的孢囊最为常见, 分别占到2种培养条件下所得全部孢囊的77.70%和95.18%。从沉积物中所得的孢囊多属于第1种类型。分析了实验条件下形成不同类型休眠孢囊的原因和比例, 同时探讨了海区沉积物中形成多种孢囊形态特征的原因, 为锥状施克里普藻孢囊的生态研究提供了依据。     

武汉鼓藻类新分类群

与原变种不同为半细胞前面观中部具有6个颗粒,颗粒间具5个圆孔纹,缢部上端具有 1个颗粒。细胞宽25～31μm,长34～42μm,厚17～21μm,缢部宽7～11μm。     

Morphological peculiarities of <Emphasis Type="Italic">Fragilidium mexicanum</Emphasis> Balech, 1988 (Dinophyta) from the Far-Eastern Seas of Russia

The vegetative cells of Fragilidium mexicanum Balech are recorded from the Far Eastern seas of Russia (Sea of Japan and Sea of Okhotsk) for the first time. Morphological study of both cultured and wild cells of F. mexicanum showed that the shape of the cell and the first (1″) and second (2″) precingular plates, the direction of the slot in plate 1″, and the shape of the anterior sulcul plate (S.a.) vary greatly. These features bear similarity to those of F. mexicanum, as well as a closely related species, F. subglobosum. The most conservative characters distinguishing the two species are the shape and size of the first and seventh postcingular plates and the shape of the posterior sulcal plate.     

Alexandrium peruvianum (Balech and Mendiola) Balech and Tangen a new toxic species for coastal North Carolina

Routine sampling of the water quality stations in the New River Estuary (Jacksonville, North Carolina, USA) during November 2004 revealed the presence of a previously unidentified dinoflagellate. Preliminary observations of its morphology suggested it to be consistent with that of Alexandrium peruvianum (Balech et Mendiola) Balech et Tangen. Observations using brightfield, epifluorescence and scanning electron microscopy confirmed the diagnostic thecal plates to be those of A. peruvanium. Clonal cultures established from cells isolated from the New River Estuary samples were also used for further studies of morphology and for the presence of toxins. Thecal morphology was consistent with that described by Balech clearly separating it from the sister species Alexandrium ostenfeldii. Three classes of toxins were detected from these cultures. An erythrocyte lysis assay (ELA) was used to confirm the presence of hemolytic toxins in A. peruvianum cultures. A cellular EC₅₀ for lysis was 1.418 × 10⁴ cells, well within the range the maximal cells densities found in the New River and more potent when compared on a cellular basis with Prymnesium parvum. Another toxin class detected in A. peruvianum cultures was the fast acting 13-desmethy C and D spirolides also produced by the sister species A. ostenfeldii. The last toxin type detected in the A. peruvianum cultures was the paralytic shellfish toxins, GTX 2, 3, B1, STX and C1,2. These findings expand the geographic range of occurrence for A. peruvianum in the U.S. to be much greater than previously considered. The morphological characters agreed with previously reported molecular data in separating A. peruvianum from A. ostenfeldii. It is also the first confirmed report that this species produces PSP toxins, spirolides and naturally occurring hemolytic substances. In light of these findings additional attention is needed for the detection of Alexandrium species in all coastal waters of the U.S. This added effort will enhance the evaluation of the relative impacts of the species to shellfish safety and bloom surveillance.     

Toxic Alexandrium peruvianum (Balech and de Mendiola) Balech and Tangen in Narragansett Bay,Rhode Island (USA)

Phytoplankton monitoring in Wickford Cove, Rhode Island, US (41°34′10.13″N, 71°26′45.76″W), located in Narragansett Bay, detected an unusual species of Alexandrium in the spring of 2009. Thecal plate analysis using brightfield and SEM microscopy revealed a plate morphology consistent with that of Alexandrium peruvianum (Balech and de Mendiola) Balech and Tangen. Molecular analyses indicated that the sequences of the SSU, ITS1, 5.8S, ITS2 and LSU through the D region of the 18S gene were similar to those of A. peruvianum from North Carolina. Toxin analyses of cells brought into culture revealed saxitoxins, gymnodimine and fast-acting spiroimines were present in the cultured clone. Saxitoxins detected included GTX 2, GTX3, B1, STX, C1 and C2. Also present in the Wickford cove isolates of A. peruvianum were 12-methyl gymnodimine and 13-desmethyl spirolide C. A. peruvianum was detected at four sites in lower Narragansett Bay: at two sites in Wickford and two sites in Jamestown, RI. A. peruvianum was observed in the spring of 2009, 2010, 2011 and 2012 at maximum abundance levels ranging from tens of cells per liter to 14,000 cells L⁻¹. The discovery of A. peruvianum in Rhode Island coastal waters, with its potential threat to public health, is notable as it appears to be an emergent bloom species globally. The presence of A. peruvianum in Narragansett Bay is the third confirmed observation of this species on the Atlantic coast of North America. Monitoring efforts in the southern New England region should incorporate measures to detect the presence of A. peruvianum toxins.     

The toxicity and intraspecific variability of Alexandrium andersonii Balech

The toxicity of Alexandrium andersonii Balech is unclear and its intraspecific variability has yet to be studied. To address these gaps in our knowledge, in the present work five strains of A. andersonii from four different localities were characterized. The results showed that despite genetic homogeneity in the 5.8-ITS (internal transcribed spacer) and large subunit (LSU) regions and similar growth rates, strains originating from different locations varied with respect to cell size, the ratios of certain pigments, and their growth patterns. Cultures of the strains grown at 20 °C were analyzed for toxicity using four different methodologies. The two officially established methods, mouse bioassay and high-performance liquid chromatography with fluorescence detection (HPLC-FLD) and post-column reaction analysis of PSP toxins, failed to show the toxicity of any strain. Strains grown at 14 °C were also negative for PSP toxins by HPLC-FLD. However, strains grown at 20 °C exhibited both a response characteristic of the presence of toxin-inhibiting voltage-gated sodium channels, as demonstrated in a neuroblastoma neuro-2a cell-based assay, as well as hemolytic activity in a sheep red blood cell assay.     

Sexual and Asexual Processes in Protoperidinium steidingerae Balech (Dinophyceae), with Observations on Life-History Stages of Protoperidinium depressum (Bailey) Balech (Dinophyceae)

ABSTRACT. A suite of morphological, histological, and molecular techniques was used to reveal for the first time division, sexuality, mandatory dormancy period of hypnozygotes, and identity of life-history stages of any Protoperidinium spp. In both Protoperidinium steidingerae and Protoperidinium depressum , asexual division occurred by eleutheroschisis within a temporary cyst, yielding two daughter cells. Daughter cells were initially round and one-half to two-thirds the size of parent cells then rapidly increased in size, forming horns before separating. Gamete production and fusion was constitutive in clonal and non-clonal cultures, indicating that both species may be homothallic. Gametes were isogamous, approximately half the size and lacking the pink pigmentation of the vegetative cells, and were never observed to feed. Gamete fusion resulted in a planozygote with two longitudinal flagella. Planozygotes of P. steidingerae formed hypnozygotes. The fate of planozygotes of P. depressum is unknown. Hypnozygotes of P. steidingerae had a mandatory dormancy period of ca. 70 days. Germination resulted in planomeiocytes with two longitudinal flagella. Nuclear cyclosis occurred in the planozygotes of P. depressum , but in the planomeiocytes of P. steidingerae . The plate tabulation and gross morphology of gametes of P. steidingerae and P. depressum differed markedly from those of vegetative cells. Thus, misidentification of morphologically distinct life-history stages and incomplete examination of thecal plate morphology in field specimens has likely led to taxonomic confusion of Protoperidinium spp. in previous studies.     

Regulation of spiroimine neurotoxins and hemolytic activity in laboratory cultures of the dinoflagellate Alexandrium peruvianum (Balech & Mendiola) Balech & Tangen

Defined experimental regimes were used to determine the effects of nutrient limitation on the toxicity of Alexandrium peruvianum in batch culture. Subsamples for cell counts and spiroimine analysis at six day intervals were used to investigate the concentrations and composition of these compounds throughout growth. An erythrocyte lysis assay for hemolytic activity was performed on cell pellets and supernatants also collected every six days over the entire growth period from all treatments. From the data, growth rates, cellular spiroimine quotas and effective concentration-fifty (EC₅₀s) for cellular and supernatant associated hemolytic activity were calculated. Phosphate limitation was identified as a key regulator of toxicity in this species, yielding maximum values of 54.1 pg cell⁻¹ for 13-desmethyl spirolide C, 96.4 pg cell⁻¹ for 12-methylgymnodimine and a potent hemolytic EC₅₀ value of 7.1 × 10³ cells. The concentrations of spiroimines detected in A. peruvianum among various treatments, in addition to a unique profile of paralytic shellfish poisoning toxins, is unique in the body of microalgal literature. Because of the multiple toxin arsenal produced by this organism, the evaluation of a single toxin clearly would have underestimated the potential virulence and significance of this clone. This study provides the first evidence that growth and toxin production of A. peruvianum are influenced by altered nutrient ratios.     

Observations on Neobursaridium gigas Balech, 1941 (Ciliata Heterotrichida)

SYNOPSIS. A large heterotrich ciliate (Family: Bursariidae) found in a papyrus swamp in Uganda was used for oxygen tension experiments by Beadle & Nilsson, 1959, under the name of Bursaria sp. This organism has now been identified as Neobursaridium gigas Balech. The morphology of the organism was studied in living and stained specimens, especially with the silver impregnation technique, and the present findings are compared to those of Balech.     

PREY SPECIFICITY AND FEEDING OF THE THECATE MIXOTROPHIC DINOFLAGELLATE FRAGILIDIUM DUPLOCAMPANAEFORME1

In summer to autumn of 2008, a recently described thecate mixotrophic dinoflagellate, Fragilidium duplocampanaeforme Nézan et Chomérat, occurred in Masan Bay, Korea, where it frequently contained bright‐orange fluorescent inclusions. Using cultures of F. duplocampanaeforme isolated from Masan Bay, we investigated feeding, digestion, and prey specificity of this mixotroph. F. duplocampanaeforme fed exclusively on Dinophysis spp. when offered a variety of prey including dinoflagellates, a raphidophyte, a cryptophyte, a ciliate, and diatoms separately. In addition, F. duplocampanaeforme had allelopathic effects on other organisms, including cell immobilization/motility decrease (in Dinophysis acuminata, D. caudata, D. fortii, D. infundibulus, Gonyaulax polygramma, Heterocapsa triquetra, and Prorocentrum triestinum), breaking of cell chains (in Cochlodinium polykrikoides), cell death (in Prorocentrum minimum), and temporary cyst formation (in Scrippsiella trochoidea). F. duplocampanaeforme engulfed whole Dinophysis cells through the sulcus. About 1 h after ingestion, F. duplocampanaeforme became immobile and shed all thecal plates. The ecdysal cyst persisted for ～7 h, during which the ingested prey was gradually digested. These observations suggest that F. duplocampanaeforme may play an important role in the Dinophysis population dynamics in the field.     

The Biology of Neobursaridium gigas Balech, 1941 (Ciliata: Heterotrichida)

SYNOPSIS. N. gigas Balech, 1941, has been found again in Uganda and cultured in the laboratory. Some features of its morphology and behavior were studied. The buccal cavity has a small pit. Trichocysts are found in the endoplasm as well as the cortex. The ciliate appears to be highly sensitive to light. Cyclops was an important predator. The size of N. gigas is very variable.