Cyst formation of the noxious red tide flagellate Chattonella marina (Raphidophyceae) in culture

Cyst formation of the noxious red tide flagellate Chattonella marina (Raphidophyceae), cultured from a cyst in sediments of Suo-nada, Japan in 1986, was investigated by culture experiments. Nitrogen limitation was effective in inducing cyst formation. After incubation in N-limited medium at 25°C with 10000 lux on a 14 h light, 10 h dark photo-cycle, cultures were subjected to six different light intensities (0 10000 lux) in order to determine effects of light intensity on cyst formation. Cysts were formed under light intensities of 1000 lux or below, but not at 10000 lux. Cysts formed in culture displayed morphological characteristics quite similar to those natural cysts observed in sediments collected from the Inland Sea of Japan. Observations on nucleus, using diaminidinophenylindole (DAPI)-staining and epifluorescence microscopy, indicated that the majority of cysts formed in culture were uninucleate, a few were binucleate. Germination of cysts was observed under adequate conditions for germination (22°C, 3500 lux with a 14 h light, 10 h dark photo-cycle) even after storage at 11°C in the dark for more than 4 mo.     

Oxygen-radical-mediated toxic effects of the red tide flagellate Chattonella marina on Vibrio alginolyticus

Toxic mechanisms of the red tide flagellate, Chattonella marina, collected in 1985 from Kagoshima Bay, Japan, were studied at the subcellular level. C. marina was found to reduce ferricytochrome c at a rate related to the concentration of plankton cells. Ca. 50% of the cytochrome c reduction was inhibited by the addition of 100 U superoxide dismutase ml^-1. These results suggest that a part of the cytochrome c reduction was caused by a superoxide anion which was extracellulary released from C. marina. Moreover, a small amount of hydrogen peroxide was detected in the C. marina suspension using the fluorescence spectrophotometric assay method. The identity of the hydrogen peroxide was confirmed by its reaction with 500 U catalase ml^-1. It is thus proposed that C. marina produces harmful active oxygen radicals and therefore exhibits a toxic effect on surrounding living organisms. In agreement with these results, C. marina strongly inhibited the proliferation of marine bacteria, Vibrio alginolyticus, in a plankton/bacteria co-culture system. The growth inhibition of bacteria caused by C. marina was related to the density and the metabolic potential of C. marina. Ruptured plankton showed no toxic effect on the bacteria. Furthermore, the toxic effect of C. marina on V. alginolyticus was completely suppressed by the addition of catalase and superoxide dismutase. In addition to these radical-scavenging enzymes, a chemical scavenger, sodium benzoate, also had a protective effect. These results suggest that oxygen radicals are important in the toxic action of C. marina.     

营养盐对海洋卡盾藻生长与产毒的影响（Effect of Some Nutrients on the Growth and Haemolytic Toxins Production of Chattonella marina）

海洋卡盾藻(Chattonella marina)是我国南方重要的有害赤潮原因种,其产生的溶血毒素、活性氧等可能是造成鱼类死亡的重要原因.在实验室条件下,研究了氮、磷和铁等营养盐对海洋卡盾藻生长及产毒的影响,以期为阐明海洋卡盾藻溶血毒素生成机制奠定基础.研究结果发现:在铁、氮、磷盐三因素四水平正交实验条件下,铁盐和氮盐是影响海洋卡盾藻生长的显著因子;氮盐和磷盐是影响海洋卡盾藻产毒的显著因子;铁盐浓度0.492mg·L-1、氮盐浓度375mg·L-1、磷盐浓度5mg·L-1时海洋卡盾藻的比生长速率最大;铁盐浓度0mg·L-1、氮盐浓度375mg·L-1、磷盐浓度0.5mg·L-1时海洋卡盾藻的产毒能力最强,即高氮、低磷条件下有利于海洋卡盾藻溶血毒素的合成.     

Possible involvement of the glycocalyx in the ichthyotoxicity of Chattonella marina (Raphidophyceae): immunological approach using antiserum against cell surface structures of the flagellate

Although the ichthyotoxic mechanism of Chattonella marina is still unknown, several lines of evidence suggest that the reactive oxygen species (ROS), such as superoxide anion (O₂^-), hydrogen peroxide (H₂O₂)_, and hydroxyl radical (·OH), produced by C. marina are involved in the mortality of fish exposed to this flagellate. Recently, we found that the cell-free supernatant prepared from C. marina, which is considered to contain the glycocalyx, showed NADPH-dependent O₂^- generation. In this study, we prepared antiserum against the crude glycocalyx of C. marina. Using indirect immunofluorescence, it was confirmed that the antiserum specifically reacted with C. marina cells. In addition to C. marina, the antiserum also reacted with other raphidophycean flagellates such as Heterosigma akashiwo, Olisthodiscus luteus, and Fibrocapsa japonica, whereas no reactivity was observed against six other flagellate species tested. These results suggest that raphidophycean flagellates have common epitopes recognized by the antiserum. Interestingly, immunohistochemical analysis of paraformaldehyde-fixed gill lamellae from yellowtail exposed to C. marina revealed that the antiserum stained the surface of gill lamellae, while no such staining pattern was observed in control gill lamellae. These results suggest that the glycocalyx may be discharged when C. marina cells are inhaled into the fishes' mouths and then come into contact with the gill surface. Based on the present results, together with our previous findings, we propose that continuous accumulation of the discharged glycocalyx on the gill surface occurs during C. marina exposure, which may be responsible for the ROS-mediated severe gill tissue damage leading to fish death.     

Bioavailable iron species in seawater measured by macroalga (Laminaria japonica) uptake

The bioavailability of iron in seawater filtered through a 0.025-m filter was investigated using ⁵⁹Fe-labeled iron uptake by the macroalga Laminaria japonica (Areschoug: Phaeophyta) (collected in the northern Japan Sea 1993) as an assay. About 80% of the iron in the 0.025-m filtered coastal seawater was soluble and/or small colloidal organically bound iron, associated with natural organic ligands forming complexes with ferric ion. After decomposition of the organic matter by ultraviolet (UV) irradiation, 55% of the iron addition [or 0.6 nM, nearly the concentration of Fe(OH) ₂ ⁺ in equilibrium with amorphous hydrous ferric oxide in seawater at pH 8.0] in the 0.025-m filtered coastal seawater was taken up by the macroalga. Since the iron concentrations in the 0.025-m filtered coastal seawater are 0.1 to 2.0 nM and only 0.6 nM of the iron is likely available to biota over 1 to 2 d, we suggest that only small amounts of bioavailable iron exist in coastal seawater not affected by inflow from land and that a significant fraction of dissolved (<0.025 ) iron occurs in forms, such as organic iron complexes, other than the simple hydroxo-complex species predicted by thermodynamic models.     

微囊藻毒素-LR对罗非鱼鳃组织活性氧自由基含量及相关抗氧化酶活性的影响

应用腹腔注射的方式,研究了微囊藻毒素MC-LR对罗非鱼（Oreochromis niloticus）鳃组织活性氧自由基含量（ROS）及过氧化氢酶（CAT）、超氧化物歧化酶（SOD）活性的影响。结果表明：在MC-LR的胁迫下,罗非鱼鳃组织产生了大量的ROS,并引起了SOD和CAT 2种抗氧化酶活性的变化。SOD和CAT在清除过量的ROS时表现出的动态变化过程不同,SOD的变化曲线是多阶段的,而CAT则表现为先诱导后恢复的趋势。SOD和CAT在变化趋势上有所不同,但均表现为明显的时间与效应的正相关关系和剂量与效应的正相关关系。这也为将鳃组织中SOD和CAT作为抗氧化生物标志物来监测微囊藻毒素对罗非鱼胁迫作用提供了一定的参考。试验为研究微囊藻毒素对罗非鱼的生态毒理学作用提供相应的资料,并为罗非鱼质量安全问题的研究打下一定的基础。     

Inhibition of the growth of micro-algae and bacteria by extracts of eelgrass (Zostera marina) leaves

The effects of the water-soluble fraction of dead leaves of the eelgrass Zostera marina L. on the growth of 8 species of micro-algae (pennate and centric diatoms, dinoflagellates, and a green flagellate) and a bacterium were studied on agar plates and in liquid culture. The extracts of leaves which had been dead from a few days to 2 wk inhibited growth and often killed cells in all test organisms. Extracts were lethal even at concentrations equivalent to as little as 0.25 mg dry leaf ml^-1, but inhibition decreased when extracts were prepared from leaves aged in the laboratory for 35 d (loss of anti-bacterial activity) or 90 d (loss of anti-algal activity). Extracts of leaves which had aged and dried several months in the field had no effect, except at very high concentrations (13 mg dry leaf ml^-1) when the lag phase of growth was prolonged several days in a culture of the chlorophyte Platymonas sp. The active fraction in eelgrass leaves may be important in controlling initial growth of micro-organisms on eelgrass detritus, and it could determine the composition and activity of the epiphytic community on living leaves.     

Abatement of sulfide generation in sewage by glutaraldehyde supplementation and the impact on the activated sludge accordingly

Hydrogen sulfide emission in sewer systems is associated with toxicity, corrosion, odour nuisance and high costs treatment. In this study, a novel method to inhibit sulfide generation from sewage by means of glutaraldehyde supplementation has been suggested and evaluated under anaerobic conditions. Different concentrations of glutaraldehyde at 10, 15, 20, 30 and 40 mg·L^-1 have been investigated. Besides, the possible impacts of glutaraldehyde supplementation on an activated sludge system and an appraisal of the economic aspects are presented as well. As observed from the experimental results, a dosage of 20 mg·L^-1 glutaraldehyde resulted in a significant decrease of the sulfide production by 70%–80% in the simulated sewage. Moreover, the impacts of additional glutaraldehyde at 20 mg·L^-1 on activated sludge, in terms of chemical oxygen demand removal and oxygen uptake rates, were negligible. From an economical point of view, the cost of the commercial glutaraldehyde products required in the operation, which was calculated on the basis of activated sulfide removal avoidance, was around €3.7–4.6 S·kg^-1. Therefore it is suggested that glutaraldehyde supplementation is a feasible technique to abate the sulfide problems in sewer systems. Yet further research is required to elucidate the optimum “booster” dosage and the dosing frequency in situ accordingly.     

Ultrastructural alterations on chloride cells of the yellowtail Seriola quinqueradiata,following exposure to the red tide species Chattonella antiqua

By means of scanning and transmission electron microscopy, the influence of the red tide species Chattonella antiqua was examined with respect to the surface ultrastructures of chloride cells of the yellowtail Seriola quinqueradiata. Conspicuous ultrastructural alterations occurred on the apical surface of these cells. The majority of chloride cells in the control gills showed an apical surface with numerous cellular extensions, while more than half the chloride cells affected by red tide organisms exhibited an apical surface with fewer and smaller extensions, a wrinkled apical surface, or a protruded apical surface. These ultrastructural alterations of chloride cell surface may be due to the partial disturbance of salinity by C. antiqua, and reflect the changes of the ion-transport function in yellowtail gills exposed to red tide water.     

Adsorption of arsenic (V) by iron (III)-modified natural zeolitic tuff

Adsorption of arsenic (V) by natural zeolitic tuff, modified with iron (III), was investigated. Also, the iron (III) adsorption characteristics by natural zeolitic tuff was evaluated. It was determined that iron (III) adsorption by starting zeolitic tuff was best represented by the Freundlich type of isotherm, having correlation coefficient (r ²) of 0.990. Arsenic (V) adsorption by iron (III)-modified zeolitic tuff followed a nonlinear type of isotherm. The best fit of the experimental data was obtained using the Langmuir–Freundlich model (r ² = 0.99), with the estimated maximum of arsenic (V) adsorption to iron (III)-modified zeolitic tuff of 1.55 mg/g.     

Impact of Arenicola marina (Polychaeta) on the microbial assemblages and meiobenthos in a marine intertidal flat

The benthic microbial food web can be responsible for a large proportion of benthic carbon cycling yet there are few data on the trophic interactions between this food web and macrobenthos. A large-scale field experiment was conducted to investigate effects of eliminating the polychaete Arenicola marina on benthic microbes (prokaryotes, heterotrophic and autotrophic protists) and metazoan meiofauna in a marine intertidal flat of the North Sea, Germany. Over a period of 2 years, quantity and composition of micro- and meiobenthos from unmanipulated sites were compared to those from sites deplete of lugworms. These grazer treatments were cross-classified with different sediment characteristics (low- and mid-intertidal areas). Lugworm removal resulted in an initial increase in abundance of prokaryotes and nanoflagellates, which became less pronounced in the second year. Ciliates were not affected quantitatively, but in the absence of lugworms, diversity and the proportion of carnivorous forms increased. Meiobenthos (nematodes, ostracods and copepods) were affected only moderately. The observed changes are probably due to a combination of release from grazing/predation pressure, changes in the species composition of higher trophic levels (namely large polychaetes) and altered environmental conditions (such as depth of the oxygenated layer and sediment grain size). Spatial differences between sites of different tidal exposure/grain size appeared to be as large as temporal differences during the 2 years following the manipulation of the system. We conclude that in intertidal sediments, indirect effects due to habitat transformation are as important as direct biological interactions (grazing pressure and competition) for the dynamics of the benthic microbial food web.     

Thermodynamic investigation of the effect of oxygen and hydrogen chloride potential upon generation and decomposition behavior of dioxins

The thermodynamic equilibrium of the C‐H‐O‐Cl system has been calculated to elucidate the generation and decomposition behavior of dioxins, and is discussed from the viewpoint of the oxygen and hydrogen chloride potential. The dioxins behavior is greatly dependent on the oxygen potential, i.e. higher temperature and oxygen potential lead to the complete decomposition of dioxins, while low hydrogen chloride potential under insufficient oxygen condition leads to an increase of multi‐chlorine displacement of dioxins.     

The effect of temperature and thermal distribution on glycolysis in two rockfish species (Sebastes)

Studies on the effects of temperature on the activities of Embden-Meyerhof (EM) glycolysis, and the hexose monophosphate shunt (HMP) in fishes have dealt mainly with exotic and/or acclimated fishes. This study reports the effects of short-term reductions in temperature on EM and HMP activity in two closely related species of temperate fishes (Sebastes spp.) and its possible relation to the thermal distribution of the species. Thermal distribution data were collected by SCUBA for S. mystinus and S. serranoides in King Harbor, Redondo Beach, California, USA. Activities of the pathways were determined in liver-tissue studies, using glucose-¹⁴C and liquid scintillation techniques following the method of Hochachka (1968) with modifications. The data were analyzed by distribution-free methods. Tissue studies indicated HMP activity in both species at lowered temperature (5°C), but only in S. serranoides at 15°C. Results indicate that S. mystinus is capable of instantaneous temperature compensation, possibly related to its tendency to occupy cold waters.     

Phytoaccumulation of iron by callus tissue of Clerodendrum indicum (L)

Clerodendrum indicum (L.) is one of the important medicinal plants, originating in India. The plant is used as medicine for bronchitis, asthma and different immunological disease. Micro-propagation is a quick method, producing a huge number of plants. Apical leaf of Clerodendrum indicum (L.) was used as an explant for callogenesis. The combinations of different plant growth regulators (2,4-D, NAA, KN) in MS (Murashige and Skoog) medium produced two types of calli: one was friable and loose and the other was green, embryogenic and compact. Heavy metals cause pollution and health hazards. The plant absorbs heavy metals, which is very useful for controlling environmental pollution. Iron is a major nutrient for both plants and animals, but hyper accumulation of iron is injurious to health. Calli were used to study the optimum conditions of iron uptake in presence of different chemical environments. The different environments were created by using different chemical reagents such as (HCL, HNO₃, H₃PO₄ and KNO₃). Iron (III) was spiked with radioactive Fe-59 and dynamics of uptake was followed by measuring radioactivity by gamma-ray spectrometer. The best condition of iron uptake corresponded to 1 mol L^?1 KNO₃ after 4 hour of equilibration.     

Promotive effect of pyridine on indole degradation by activated sludge under anoxic conditions

Batch experiments were carried out to investigate the promotive effect of pyridine on indole degradation under denitrifying conditions. The seed sludge was obtained from a local coal-coking wastewater treatment facility and was acclimated in the laboratory. Indole and pyridine were supplemented to the synthetic wastewater at different ratios. The optimum ratio of chemical oxygen demand (COD) to nitrate (C/N) was 8.4–8.9 for both denitrification and indole and pyridine degradation. At a temperature of 28°C and pH of 7.0–7.5, the nitrate reductase activity (NRA) was in the best state. The addition of pyridine could promote NRA and the degradation of indole. When the initial concentration of indole was 150 mg/L, the concentration ratio of indole to pyridine was in the range of 1–10. Under optimum C/N conditions, the degradation of indole could be described with pseudo-zero-order kinetics. There was no accumulation of nitrite during the reaction. When the concentration ratio of pyridine to indole was less than 0.25 with an increase in the pyridine proportion, there were more significant augment rates for NRA and the degradation of indole than the situation when the concentration ratio was more than 0.25.     

Enhancement of As(V) adsorption onto activated sludge by methylation treatment

Biosorption properties of arsenate [As(V)] onto activated sludge were investigated in batch systems. The adsorption of As(V) onto sludge increased from 23 to 266 μg/g dry weight through the methylation of the activated sludge. This increase resulted from neutralization of carboxylic groups via the methylation process. The pH effect of As(V) uptake was also investigated and As(V) adsorption by methylated sludge decreased significantly at high pH (pH > 11) due to competition between As(V) and OH⁻ ions for binding sites distributed on sludge surfaces. In contrast, low pH favored As(V) adsorption by methylated sludge because of the elevated quantities of positively charged functional groups. The results suggest that methylated activated sludge may provide promising applications for the simultaneous removal and separation of As(V) from aqueous effluents.     

Removal of chromium(VI) by ZnCl2 activated coir pith carbon

The adsorption of chromium(VI) onto ZnCl₂ activated carbon developed from coir pith was investigated to assess the possible use of this adsorbent. The influence of contact time, adsorbent dose, Cr(VI) concentration, pH and temperature were investigated. The two theoretical adsorption isotherms, namely, Langmuir and Freundlich were used to describe the experimental results. The Langmuir adsorption capacity (Q ₀) was found to be 120.5?mg Cr(VI) per g of the adsorbent. The adsorption followed the second-order kinetics and was found to be maximum at pH 2.0. The pH effect and the desorption studies showed that ion exchange mechanism might be involved in the adsorption process. The effects of foreign ions such as chloride, sulphate, phosphate, selenite, molybdate, nitrate and perchlorate on the removal of Cr(VI) have been investigated. The removal of Cr(VI) from synthetic ground water was also tested. The results show that ZnCl₂ activated coir pith carbon is effective for the removal of Cr(VI) from water.     

盐京九号水稻秸秆对铜绿微囊藻(Microcystis aeruginosa)的抑制作用

以盐京九号水稻为材料,研究经高温灭菌或物理粉碎后的水稻秸秆微细颗粒对铜绿微囊藻(Microcystisaeruginosa)生长的抑制及其作用机制.结果表明,当藻液中水稻秸秆添加量≥10g·L-1时,高温灭菌与未灭菌的水稻秸秆同样能抑制微囊藻生长.灭菌稻秆试验组藻细胞的呼吸速率和SOD活性都呈现出先上升后迅速下降的趋势;通过有机溶剂(石油醚、乙酸乙酯和乙醚)萃取稻秆水浸液,用滤纸片法在固体培养基上所做的抑藻试验结果表明,乙醚提取液有明显的抑藻圈.提示抑藻效应是水稻秸秆中某些化学物质溶出而产生的作用,与微生物的活动无关.进一步分离鉴定乙醚提取液结果显示,浸提液中有机酸类和酚类物质有显著的抑藻效应.水稻秸秆微粒的粒径越小,产生的抑藻效应出现得越早,也越明显,这可能还与秸秆微粒的吸附作用有关.     

Mo6+对活性污泥系统反硝化性能的影响

采用对比实验的方法,在不同初始硝酸盐氮和COD质量浓度的条件下,对人工模拟的活性污泥系统投加不同质量浓度的Mo6 ,以此考察Mo6 对活性污泥系统反硝化性能的影响。结果发现,Mo6 的质量浓度小于5mg?L-1以下时能够促进硝酸盐氮的去除,质量浓度为1mg?L-1时促进效果最好。Mo6 的质量浓度在4mg?L-1以下时,可以使COD去除效率增加,质量浓度为2mg?L-1时有最佳促进作用。Mo6 的质量浓度小于1g?L-1时可提高反硝化活性污泥的TTC-脱氢酶活性,过高质量浓度则表现出抑制作用。综合Mo6 的对硝酸盐氮和COD去除的影响规律以及对活性污泥TTC-脱氢酶活性的影响,认为Mo6 在质量浓度为1mg?L-1时对反硝化的促进作用最强。