STUDIES ON MIXED MASS CULTIVATION OF ANABAENA SPP (NITROGEN-FIXING BLUE-GREEN-ALGAE, CYANOBACTERIA) ON A LARGE-SCALE

Studies on mixed mass cultivation of Anabaena spp. on a large scale (5170 m2) were conducted continuously for 3 years. Under the continental monsoon climate in northern subtropics (30-degrees-N, 115-degrees-E), 7-11 g dry weight m-2 day-1 of microalgal biomass on average was harvested in simple plastic greenhouses in the effective growth days during the warmer seasons. The maximum productivity was 22 g m-2 day-1 in the middle of summer. Observations on the productive properties of strains of Anabaena spp. indicated that they were different from and could compensate for each other in their productivities and adaptations to the seasonal changes. With different lining materials (PVC sheets, concrete, sand and soil) in the culture ponds, no significant variation of productivity was found, but bubbling with biogas in the middle of the day and the application of some growth regulating substances (2,4-D, NaHSO3 and extracts of oyster mushroom spawn) was able to improve the production. The cost of microalgal biomass in this way was around 0.75-1.0 US dollar(s) per kilogram.

Mechanical and thermal induced chain conformational transformations in syndiotactic polypropylene (sPP)

Stretching a stacked sPP lamellar morphology at room temperature leads to a mechanical induced transformation from the (t(2)g(2))(2) (helical) into the (tttt) (zigzag) chain conformation of the polymer. The so prepared samples exhibit after annealing above 80 degreesC a thermal induced retransformation into the cell I and cell III crystal structure of the helical chain conformation. The mechanical induced chain conformational transformation as well as the thermal induced retransformation was studied by means of transmission electron microscopy and electron diffraction. (C) 2001 Kluwer Academic Publishers.

Epitaxial crystallization of sPP on the (100) lattice plane of HDPE crystals

Crystallization behavior of syndiotactic polypropylene(sPP) on the (100) lattice plane of high-density polyethylene(HDPE) crystals was studied by means of transmission electron microscopy and electron diffraction. The results indicate that sPP crystals can grow epitaxially on the (100) PE lattice plane with their chain directions +/-37 degrees apart from the chain direction of the HDPE substrate. The contact planes are (100) lattice planes for both polymers. This kind of epitaxy is explained in terms of parallel alignment of HDPE chains along the rows formed by the {CH3, CH2,CH3} groups in the (100) lattice plane of the sPP crystals. This implies that in the epitaxial crystallization of sPP with fiber oriented HDPE substrate, not only the (110) but also the (100) HDPE lattice planes can act as the oriented nucleation sites. Furthermore, according to the poor matching between HDPE chains in the (100) lattice plane and the {CH3, CH2, CH3} group rows in the (100) lattice plane of the sPP crystals, it is concluded that the geometric matching is not the only controlling factor for the occurrence of polymer epitaxy.

Crystallization of syndiotactic polypropylene (sPP) from oriented melts

The influence of the syndiotacticity on the crystallization behaviour of syndiotactic polypropylene (sPP) has been investigated. The syndiotacticity has been measured by C-13-NMR spectroscopy and the phase formation has been observed by electron diffraction of oriented samples. It is shown that the crystal phase formation depends strongly on the perfection of the tacticity of the macromolecules.

Epitaxial crystallization in the sPP/nylon-12 semicrystalline polymer system

The epitaxial crystallization behaviour of syndiotactic polypropylene (sPP) on highly oriented nylon-12 substrates has been investigated by means of transmission electron microscopy. The results obtained from bright field electron microscopy and electron diffraction indicate that sPP crystals grow epitaxially on the oriented nylon-12 substrate with their c-axes +/- 37 degrees apart from the chain axis of the nylon-12 substrate. The contact planes of the sPP crystals are the (100) lattice planes. Moreover, the epitaxial crystallization of nylon-12 on highly oriented sPP substrates from a dilute solution in cyclohexanone has also been studied using optical microscopy. The results show that the nylon-12 crystals grow epitaxially on the oriented sPP substrate with the oriented nylon-12 lamellae forming large, anisotropic domains. Copyright (C) 1996 Elsevier Science Ltd.

织纹螺（Nassarius spp.）体内河豚毒素来源的研究

近几十年来，国内沿海地区频繁发生食用织纹螺中毒事件，并导致数十人死亡，这一问题得到了政府相关部门的高度重视。但是，由于织纹螺毒性变化很大，毒素来源不清楚，因此很难预测食用织纹螺中毒事件的发生，这在很大程度上限制了对食用织纹螺中毒事件的有效监测和管理。目前，对于中国沿海有毒织纹螺体内河豚毒素（tetrodotoxin, TTX）的来源还未见过系统研究。本文选取中国沿海常见的半褶织纹螺（Nassarius semiplicatus）、纵肋织纹螺（N. variciferus）和拟半褶织纹螺（N. semiplicatoides sp. nov.）作为实验对象，从毒素的微生物来源与食物链来源这两个角度分别展开研究，以探讨织纹螺体内 TTX 的可能来源，为提出相应的预防管理措施提供科学依据。 首先，我们先后从曾发生过中毒事件的江苏盐城和连云港采集了织纹螺样品，通过小鼠生物测试法和液-质联用分析技术（LC-MS），对织纹螺的毒性和毒素组成进行了测试和分析，分离培养了织纹螺体内及其生活环境中的细菌，应用河豚毒素单克隆抗体酶联免疫检测方法（ELISA）对细菌的产毒情况进行了测试，并通过 16S 核糖体（rRNA）部分基因序列测定对细菌种类进行了初步的分析。研究发现，采自江苏盐城和连云港的半褶织纹螺的毒性分别约为 2 MU/g 和 200 MU/g 组织，体内的毒素成分是河豚毒素及其同系物。从盐城的半褶织纹螺及其生活环境分离的菌株中随机挑出 14 个菌株中，9 个菌株河豚毒素检测结果呈现阳性。从连云港高毒性半褶织纹螺消化腺中分离到的 45 个菌株中，阳性菌株有 21 个。但是，有毒菌株毒素含量较低，毒素含量范围是 15-184ng/g。通过 16S rDNA 部分序列的测序结果发现，大部分有毒菌株与弧菌属（Vibrio）的细菌在遗传序列信息上比较相近。其余有毒菌株分别与希瓦氏菌属（Shewanella）、海单胞菌属（Marinomonas）、黄杆菌属（Tenacibaculum）、动性菌属（Planococcus）、发光杆菌属 (Photobacterium)和气单胞菌属（Aeromonas）的遗传序列比较相近。其中与海单胞菌属、动性菌属和发光杆菌属亲缘关系较近的产毒细菌是首次报道。这一研究表明织纹螺体内及其生活环境中的存在产河豚毒素的细菌，但由于产毒素的量较低，因此可能在织纹螺体内河豚毒素的产生和累积过程并不发挥主要作用。 织纹螺作为一类腐食性的海洋动物，也有可能通过进食含有河豚毒素的生物而累积河豚毒素。对此，我们开展了高毒性半褶织纹螺的室内培养实验，以及河豚毒素在不同种类织纹螺体内的累积和排出的模拟实验，并定期采样，通过液相色谱与串联质谱联用技术（LC-MS/MS）对织纹螺体内河豚毒素及其同系物的含量变化情况进行了分析。室内培养实验发现，从连云港赣榆县采集的高毒性半褶织纹螺，在实验初期，体内毒素含量呈下降的趋势，但从 7月上旬开始，毒素含量突然快速上升，与连云港赣榆县野外采集的织纹螺的毒素含量表现出相似的变化趋势。河豚毒素在不同种织纹螺体内的累积和排出的模拟实验发现，通过投喂高毒性的河豚鱼肝脏（毒性为5×103 MU/g），纵肋织纹螺在一段时间内能够快速累积少量的河豚毒素。当停止投喂有毒河豚鱼肝脏后，毒素含量会快速下降。而在曾导致中毒事件的拟半褶织纹螺中，投喂有毒河豚鱼的肝脏后，其体内毒素含量只有缓慢增加。但在投喂无毒的河豚鱼肝脏后，其毒性却出现了快速增加的现象，这与该地区野外样品的毒性变动状况类似。这些发现显示高毒性半褶、拟半褶织纹螺体内的河豚毒素应当不是食物链累积的结果，而可能是由其自身产生。并且，毒素含量的变化具有一定的生物节律，有可能与产卵、繁殖等自然节律相关。 通过对半褶、纵肋和拟半褶织纹螺的研究工作可以认为，产河豚毒素的细菌不是织纹螺体内河豚毒素的主要来源，并且毒素也不是来自其摄食的食物，推测可能主要是由织纹螺自身产生。织纹螺所表现出的河豚毒素含量的季节性变化，极有可能与产卵、繁殖等自然节律相关，这些发现为预防和管理食用织纹螺中毒事件提供了科学依据。但是，本研究并未完全阐明织纹螺体内河豚毒素的来源，对于织纹螺体内河豚毒素的确切来源以及河豚毒素的代谢和转化机制，还有待于更加深入地研究工作。

PCR-DGGE analysis of intestinal bacteria and effect of Bacillus spp. on intestinal microbial diversity in kuruma shrimp (Marsupenaeus japonicus)

In this study, the intestinal microbiota of kuruma shrimp (Marsupenaeus japonicus) was examined by molecular analysis of the 16S rDNA to identify the dominant intestinal bacteria and to investigate the effects of Bacillus spp. on intestinal microbial diversity. Samples of the intestines of kuruma shrimp fed normal feed and Bacillus spp. amended feed. PCR and denaturing gradient gel electrophoresis (DGGE) analyses were then performed on DNA extracted directly from the guts. Population fingerprints of the predominant organisms were generated by DGGE analysis of the universal V3 16S rDNA amplicons, and distinct bands in the gels were sequenced. The results suggested that the gut of kuruma shrimp was dominated by Vibrio sp. and uncultured gamma proteobacterium. Overall, the results of this study suggest that PCR-DGGE is a possible method of studying the intestinal microbial diversity of shrimp.