Experimental studies on dimethylsulfide (DMS) and dimethylsulfoniopropionate (DMSP) production by four marine microalgae

The production of dimethylsulfide （DMS） and dimethylsulfoniopropionate （DMSP） by marine microalgae was investigated to elucidate more on the role of marine phytoplankton in ocean-atmosphere interactions in the global biogeochemical sulfur cycle.Axenic laboratory cultures of four marine microalgae–Isochrysis galbana 8701,Pavlova viridis,Platymonas sp.and Chlorella were tested for DMSP production and conversion into DMS.Among these four microalgae,Isochrysis galbana 8701 and Pavlova viridis are two species of Haptophyta,while Chlorella and Platymonas sp.belong to Chlorophyta.The results demonstrate that the four algae can produce various amounts of DMS（P）,and their DMS（P） production was species specific.With similar cell size,more DMS was released by Haptophyta than that by Chlorophyta.DMS and dissolved DMSP （DMSPd） concentrations in algal cultures varied significantly during their life cycles.The highest release of DMS appeared in the senescent period for all the four algae.Variations in DMSP concentrations were in strong compliance with variations in algal cell densities during the growing period.A highly significant correlation was observed between the DMS and DMSPd concentrations in algal cultures,and there was a time lag for the variation trend of the DMS concentrations as compared with that of the DMSPd.The consistency of variation patterns of DMS and DMSPd implies that the DMSPd produced by phytoplankton cells has a marked effect on the production of DMS.In the present study,the authors’ results specify the significant contribution of the marine phytoplankton to DMS（P） production and the importance of biological control of DMS concentrations in oceanic water.

Effect of Benzo[a]pyrene on Detoxification and the Activity of Antioxidant Enzymes of Marine Microalgae

The objective of this study was to examine the effect of benzo[a]pyrene(Ba P) on the detoxification and antioxidant systems of two microalgae,Isochrysis zhanjiangensis and Platymonas subcordiformis.In our study,these two algae were exposed to Ba P for 4 days at three different concentrations including 0.5 μg^(L-1)(low),3 μg^(L-1)(mid) and 18 μg^(L-1)(high).The activity of detoxification enzymes,ethoxyresorufin O-deethylase(EROD) and glutathione S-transferase(GST) increased in P.subcordiformis in all Ba P-treated groups.In I.zhanjiangensis,the activity of these two enzymes increased at the beginning of exposure,and then decreased in the groups treated with mid-and high Ba P.The activity of antioxidant enzyme superoxide dismutase(SOD) increased in I.zhanjiangensis in all Ba P-treated groups,and then decreased in high Ba P-treated group,while no significant change was observed in P.subcordiformis.The activity of antioxidant enzyme catalase(CAT) increased in I.zhanjiangensis and P.subcordiformis in all Ba Ptreated groups.The content of malondialdehyde(MDA) in Isochrysis zhanjiangensis increased first,and then decreased in high Ba P-treated group,while no change occurred in P.subcordiformis.These results demonstrated that Ba P significantly influenced the activity of detoxifying and antioxidant enzymes in microalgae.The metabolic related enzymes(EROD,GST and CAT) may serve as sensitive biomarkers of measuring the contamination level of Ba P in marine water.

COMPARATIVE STUDIES OF INORGANIC CARBON UTILIZATION IN EMILIANIA HUXLEYI AND SOME NON-CALCIFYING MARINE MICROALGAE

Inorganic carbon utilization in the non calcifying marine microalgae, Nannochloropsis oculata, Phaeodactylum tricornutum and Porphyridium purpureum was compared with high and low calcifying strains of Emiliania huxleyi grown in artificial seawater medium aerated with either air (0.03% V/V CO 2) or CO 2 free air. For high calcifying strain of E. oculata and P. tricornutem , similar growth patterns were observed in air and CO 2 free air grown cultures. P. purpureum showed a less final cell density in CO 2 free air than in air grown culture. However, low calcifying strain of E. huxleyi was able to grow only in air grown culture, but not in CO 2 free air grown culture. Measurements of alkalinity, pH, concentration of dissolved inorganic carbon (DIC) and free CO 2 showed different patterns of DIC utilization. With N. oculata, P. tricornutum and P. purpureum the pattern of DIC utilization was characterized by an increase of pH and a decrease of DIC but a constant alkalinity in the cultures aerated with air or CO 2 free air, suggesting that bicarbonate utilization was concomitant with an efflux of OH -. Both alkalinity and pH were maintained rather constant in air grown culture of low calcifying strain of E. huxleyi, suggesting that diffusive entry of CO 2 could meet the requirement of DIC for its photosynthesis and growth. High calcifying strain of E.huxleyi , however, showed a pattern of decrease of alkalinity and DIC but an almost constant pH , indicating that bicarbonate was the major form of inorganic carbon utilised by this organism and bicarbonate uptake is unlikely to be accompanied by an efflux of OH -. The final pH values reached by N. oculata, P. tricornutum and P. purpureum in a closed system were 10.75, 10.60 and 9.85 respectively, showing that bicarbonate utilisation is concomitant with an efflux of OH . While the final pH of 8.4 in high calcifying E. huxleyi suggests that bicarbonate utilization was not accompanied by an efflux of OH -.

Competition within the marine microalgae over the polar dark period in the Greenland Sea of high Arctic

With the onset of winter, polar marine microalgae would have faced total darkness for aperiod of up to 6 months. A natural autumn community of Arctic sea ice microalgae was collected for dark survival experiments from the Greenland Sea during the ARKTIS-XI/2 Expedition of RV Po-larstem in October 1995. After a dark period of 161 days, species dominance in the algal assemblage have changed from initially pennate diatoms to small phytoflagellates (<20 μm). Over the entire dark period, the mean algal growth rate was - 0.01 d-1. Nearly all diatom species had negative growth rates, while phytoflagellate abundance increased. Resting spore formation during the dark period was observed in less than 4.5% of all cells and only for dinoflagellates and the diatom Chaetoceros spp. We assume that facultative heterotrophy and energy storage are the main processes enabling survival during the dark Arctic winter. After an increase in light intensity, microalgal cells reacted with fast growth within days. Phytoflagellates had the highest growth rate, followed by Nitzschia frigida. Further investigations and experiments should focus on the mechanisms of dark survival (mixotrophy and energy storage) of polar marine microalgae.

Open Mass Cultures of Marine Microalgae for Biodiesel Production: Laboratory Approach to Study Species Competition in Mixed Cultures

Since the interspecific competition represents a critical issue for biodiesel production in open mass cultures of microalgae, laboratory studies aimed to select competitive species receive significant interest. In this work, a laboratory approach based on the method of multisizing cell counting was developed to monitor the competition course of single algal species grown in mixed cultures. Two marine microalgae, Dunaliella tertiolecta and Phaeodactylum tricornutum, were used to set-up an induced-contamination assay, with the aim to test their competitive capabilities. The evaluation of the oil production by dried biomass extraction was coupled to the growth assay to investigate the effect of competition on oil yield. The adopted experimental approach revealed effective as a method to selectively measure the algal growth of single species in mixed cultures, enabling to evaluate the competitive properties of Dunaliella outgrowing the contaminant species Phaeodactylum. At the end of the experiment the dominant species contributed more than 90% to the total biomass, while no loss of oil production was observed, the oil yield in the mixed being even higher than in the unialgal culture (3.15 vs. 2.28 mg). The outcomes of the induced competition suggest the use of Dunaliella as a competitive oil-producer species and especially support the potential of the experimental approach to be used for preliminary screening to drive species selection for open mass cultures.

Effects of pretreatment procedures on fatty acid composition and stable carbon isotopes in the marine microalga Isochrysis zhanjiangenisis

This study aims to quantify the effects of different pretreatment methods on the stable carbon isotope values of fatty acids in marine microalgae（Isochrysis zhanjiangenisis）.To identify the effects of sample preparation on theδ^（13）C value and the fatty acid composition,we examined eight types of pretreatment methods including：（a） drying the sample followed by direct methyl esterification using HCl-CH3OH;（b） drying the sample followed by direct methyl esterification using H2SO4-CH3OH;（c） drying the sample by ultrasonic extraction and methylesterification using HCl-CH3OH;（d） drying the sample by ultrasonic extraction and methyl-esterification using H2SO4-CH3OH;（e） fresh sample followed by direct methyl-esterification using HCl-CH3OH;（f） fresh sample followed by direct methyl-esterification using H2SO4-CH3OH;（g） fresh sample with ultrasonic extraction followed by methyl-esterification using HCl-CH3OH,and（h） fresh sample with ultrasonic extraction followed by methylesterification using H2SO4-CH3 OH.The results show that the δ^（13）C values from Groups a-e,g and h fluctuated within 0.3‰,and the δ^（13）C values of Group f were approximately 0.7‰ lower than the other seven groups.Therefore,the different sample pretreatment methods used towards the extraction of fatty acids from marine microalgae may result in different results regarding the stable carbon isotope ratios,and if necessary a correction should be applied.

Marine ecological risk assessment for the herbicide sulfometuron-methyl based on species sensitivity distribution approach

In recent years,herbicide sulfometuron-methyl(SM)has been used to kill the invasive plant Spartina alterniflora in some coastal areas of China,which may lead to the toxic effects on non-target marine organisms.The 96-h median effective concentrations(96-h EC50)of SM on six species of marine microalgae were measured in growth inhibition tests,and were then compared with other published toxicity data,based on which a method of species sensitivity distribution(SSD)was built to estimate the hazardous concentration of SM for 5%of species(HC5)and potentially affected fraction(PAF)for a certain concentration.Results indicate that SM exhibited a high toxicity to two species of green algae(Chlorella pacifica and Dunaliella salina)with a 96-h EC50 of 0.11 and 0.13 mg/L respectively,had a medium toxicity to two species of golden algae(Diacronema viridis and Isochrysis galbana)with a 96-h EC50 of 14.24 and 21.48 mg/L respectively,and showed a low toxicity to two species of diatoms(Skeletonema costatum and Phaeodactylum tricornutum)with a 96-h EC50 of 148.99 and>100 mg/L,respectively.The estimated values of HC5 and the predicted no-effect concentrations(PNEC)for SM were 0.077 and 0.015 mg/L,respectively.According to the current dosage for killing S.alterniflora in tidal flats in Fujian Province,China,SM entering the sea by spraying might cause the acute injury or death of 14%of marine species.This hazard could last for about a month for those sensitive species.Therefore,on the premise of inhibiting the growth of this invasive plant,the dosage of SM should be reduced as much as possible to avoid severe damage to the marine ecosystem.The results provide a valuable information for marine ecological risk assessment on SM and for marine environmental management.

Antibacterial screening of silver nanoparticles synthesized by marine micro algae

Objective:To explore the biosynthesis of silver nanoparticles synthesized by marine microalgae. Methods:Marine microalgae was collected from Central Marine Fisheries Research Institute (CMFRI,tuticorin) and cultured in the lab.Silver nanoparticles synthesis were observed in normal and microwave irradiated microalgae and screened against human pathogens for the presence of antimicrobials.Results:The presence of silver nanoparticle was confirmed by UV-Visible spectroscopy at 420 nm by the presence of plasmon peak.Further confirmation was done by scanning electron microscope(SEM).Conclusions:These results not only provide a base for further research but are useful for drug development in the present and future.

The Effect of Nitric Oxide on the Growth of Marine Phytoplankton

The incubation experiments of Skeletonema costatum, Dicrateria zhanjiangensis nov. sp., and Platymonas subcordiformis, and those of Emiliania huxleyi were carried out in the Marine Physical Chemistry Laboratory in Ocean University of China and in the Marine Organic Geochemistry Laboratory in the University of Georgia respectively. Nitric oxide was added into the media when these marine microalgae were growing. We found the grovth of these four microalgae were promoted or inhibited when nitric oxide of different concentrations was added one or two times each day during the cultivation process. The results are consistent with the influence of nitric oxide on the growth of high plants. The results show that nitric oxide may be a new factor of regulation and control for the phytoplankton growth in seawater.

Diversity and community structure of eukaryotic microalgae in surface sediments in the central Bohai Sea,China,based on a metabarcoding approach

Sediment samples were collected at 17 stations in the central Bohai Sea,China,and the diversity and community structure of eukaryotic microalgae were assessed by metabarcoding the V4 region of 18S r DNA.A total of 930 operational taxonomic units(OTUs)were detected for microeukaryotes,including 98algal OTUs.The algal communities comprised 42 genera belonging to 19 classes of six phyla,and they were dominated by chrysophytes and dinoflagellates.Dinoflagellates were also the most diverse microalgal group.The nano-sized dinoflagellates Biecheleria halophila and Azadinium trinitatum occurred abundantly in the study area;however,they have not been reported previously,as they may be overlooked or misidentified in light microscopy.Many pico-sized chlorophytes were detected in the sediment samples.Sixteen of the detected OTUs were assigned to potentially harmful and/or bloom-forming microalgae,suggesting some potential risks of harmful algal blooms in the central Bohai Sea.The capacity of metabarcoding to detect morphologically cryptic and small species makes this method a sufficiently sensitive means of detection for assessing eukaryotic microalgae in sediments.

基于改进UCTransNet的海洋微藻图像分割模型

海洋微藻是海洋生态系统的基石,对其进行分割识别可以监测海洋水质并防治藻华。UCTransNet是使用Transformer模块替代UNet中跳跃连接模块的分割模型,但UCTransNet过于重视图像的通道信息而忽略了图像的空间信息。针对此情况,提出一种将空间与通道融合的注意力机制,并将其加入UCTransNet中,得到CSAM-UCTransNet。该模型加强了编码器与译码器之间的联系。实验表明,CSAM-UCTransNet对海洋微藻样本的分割精度相较于UCTransNet提升了4.88%。与UNet、Attention-UNet、UNet++等分割算法相比,该模型分割精度更高,对细节的处理效果更好。

UV-B Radiation Suppresses the Growth and Antioxidant Systems of Two Marine Microalgae, Platymonas subcordiformis （Wille） Hazen and Nitzschia closterium （Ehrenb.） W. Sm

Abstract: This study investigated whether increased solar UV-B radiation (280–315 nm) could suppress the growth of marine microalgae through effects on their antioxidant systems. Two marine microalgae species, Platymonas subcordiformis (Wille) Hazen and Nitzschia closterium(Ehrenb.) W. Sm, were exposed to a range of UV-B radiation and both showed reductions in their growth rates, and the chlorophyll a (Chl a) and carotenoid (Car) contents when UV-B radiation dose increased. Superoxide anion radical (O2-.) production and the concentration of hydrogen peroxide (H2O2) and malodiadehyde (MDA) also increased with the increasing of UV-B radiation. Antioxidant systems, non-enzymic components (Car and glutathione content) and enzymic components (superoxide dismutase (SOD) and catalase (CAT) activity), decreased as a result of enhanced UV-B radiation. When the exogenous glutathione (GSH) was added, the effects of UV-B radiation on the growth of the two species were alleviated. These results suggest that enhanced UV-B radiation suppressed the antioxidant systems and caused some active oxygen species to accumulate, which in turns retarded the development of the marine microalgae.

Effect of TiO_2 nanoparticle aggregation on marine microalgae Isochrysis galbana

TiO2 nanoparticles（NPs） could adversely impact aquatic ecosystems. However, the aggregation of these NPs could attenuate this effect. In this work, the biological effects of TiO2 NPs on a marine microalgae Isochrysis galbana were investigated. The aggregation kinetics of TiO2 NPs under different conditions was also investigated to determine and understand these effects. Results showed that, though TiO2 NPs had no obvious impact on the size and reproducibility of algal cells under testing conditions, they caused a negative effect on algal chlorophyll, which led to a reduction in photosynthesis. Furthermore, fast aggregation of TiO2 NPs occurred under all conditions, especially at the pH close to the p Hzpc. Increasing ionic strength and NP concentration also enhanced the aggregation rate.The aggregation and the following sedimentation of TiO2 NPs reduced their adverse effects on I. galbana.

海藻生物制药推动海洋强国建设研究进展

海藻细胞内含有丰富的多糖,是未来海洋生物医药研究领域的前沿热点。目前,海藻多糖积累主要聚焦于大型海藻,与大型海藻相比,海洋微藻具有种类繁多、操作简单、测定精准等优势,是亟待挖掘的有潜力的海洋生物医药的种质资源。文章从药用海藻资源的开发、药用海藻资源的培养、海藻生物制药的环境保障分析入手,分析海洋生物制药存在的瓶颈,挖掘切实可行的解决途径,提出“海洋微藻生物制药全链条”新理念,为助推海洋强国建设、提升海洋生物医药的经济效益和市场竞争力奠定基础。

三嗪除草剂扑草净对海洋浮游植物群落的影响——基于对微藻种群动态、种间竞争和光合能力的分析

以三角褐指藻(Phaeodactylum tricornutum)、海水小球藻(Chlorella pacifica)和球等鞭金藻(Isochrysis galbana)为研究对象,设置单藻培养组和双藻1:1(细胞密度比)共培养组,研究了扑草净对3种海洋饵料微藻种群动态、光合能力和种间竞争的影响.结果表明,扑草净对三角褐指藻、海水小球藻和球等鞭金藻的96h-EC50值分别为为7.65,16.67,4.11μg/L,扑草净对3种微藻的毒性大小为:球等鞭金藻>三角褐指藻>海水小球藻;扑草净通过抑制微藻的光合作用使微藻种群数量显著降低;扑草净暴露减弱了海水小球藻对三角褐指藻的竞争抑制且暴露于环境浓度(0.5和5μg/L)扑草净12d后便可导致海水小球藻和球等鞭金藻共培养体系中的敏感种球等鞭金藻全部消亡,最终使得种间作用向着更有利于硅藻的方向发展.研究结果为评估三嗪类除草剂扑草净的海洋生态风险提供了理论支撑.

海洋微藻的生态化学计量学与微藻增养殖氮磷化学计量条件的应用与研究进展

总结关于元素化学计量的文献基础上,回顾了化学计量的起源和发展,从不同层面介绍了海洋浮游植物的生态化学计量学,阐述了几种应用较为广泛的生态化学计量模型,同时介绍了氮磷化学计量学在海洋微藻增养殖中的应用,分析了常用培养基配方中氮和磷的组成特征。该领域的研究已证明,浮游植物的生长率控制与环境因子、细胞元素组成及其培养基的组成有着密切的关系。重要的是氮磷比不是决定浮游植物增殖的唯一因素,而应该同时考虑氮和磷绝对浓度的影响,特别是在解决营养限制问题时尤其需要综合考量。

海洋微藻碳酸酐酶研究进展

海洋藻类通过光合作用同化CO_(2)和H_(2)O,贡献了全球初级生产力的46%,对其固碳机制的探索研究意义重大.受限于水环境中的无机碳理化性质以及固碳关键酶对底物CO_(2)的低亲和力,海洋光合藻类演化出碳浓缩机制(CCM)以提高固碳效率.碳酸酐酶(CA)是CCM中的关键酶,能催化HCO_(3)-与CO_(2)的可逆转化.得益于近年来藻类基因组学和转录组学的快速发展,藻类CA的鉴定、生理特性及其与CCM的关系研究深入到分子水平,人们基于多样性特征,包括不同亚细胞定位、基因序列多样性、蛋白结构多样性以及功能多样性,将CA划分为不同家族.本文综述不同光合微藻的CA基因家族多样性、生理活性及其在CCM中作用的研究进展,提出未来需进一步拓展CA基因家族在不同微藻CCM中生理功能多样性与环境响应机制的研究,包括:加强海水升温与酸化、大洋环境中金属离子共限制对微藻CA活性及CCM的影响等方面的研究;在具有重要生态意义与珊瑚共生的虫黄藻中深入开展CA研究;分析CA结合不同金属辅因子的生态学影响;并对微藻CA在全球气候变化以及“碳中和”背景下的应用前景进行展望.

有机磷农药对海洋微藻致毒性的生物学研究 Ⅵ.对硫磷对4种海洋微藻的毒性效应

本文报道有机磷农药——对硫磷对４种海洋微藻的毒性效应。实验结果显示：（１）对硫磷对４种海洋微藻有较高的毒性，对藻细胞的生长和分裂有严重的抑制效应。（２）４种海洋微藻对对硫磷的耐受力有一定的差异，按由大到小的顺序依次为新月菱形藻＞盐藻＞金藻＞扁藻。（３）对硫磷能够引起叶绿素ａ的降解，随着浓度的提高，藻细胞叶绿素ａ的含量呈下降趋势。

低浓度蒽对两种海洋微藻生长的兴奋效应

以两种海洋微藻金藻 870 1(Isochrysisgalbana 870 1)和骨条藻 (Skeletonemacostatum )为材料 ,研究了低浓度蒽 (1 5～ 6 μg·L-1)对两种海洋微藻生长的影响 .结果表明 ,在一定浓度蒽作用下 ,两种微藻的生长均呈现出较明显的“兴奋效应” ,细胞密度有所增加 ,但达到最佳刺激作用的浓度与时间有所不同 .在刺激浓度作用下 ,两种微藻体内的蛋白质、叶绿素a(chla .)、类胡萝卜素 (car.)含量都有所增加 ,其变化趋势与细胞密度的变化类似 .在整个实验过程中 ,微藻体内清除活性氧的关键性酶超氧化物歧化酶 (SOD)始终处于较高水平 。

三种抗生素对几种海洋微藻叶绿素a含量影响的初步研究

实验室条件下,研究了氯霉素(chloramphenicol,Cm)、遗传霉素(geneticin G418)、青霉素(penicillin)对三种常见海洋饵料微藻:小球藻(Chlorella vulgaris Beij.),金藻8701(Isochrysis galbana Parke 8701.)和小新月菱形藻(Nitzschia closteri-um Ehr.)叶绿素a含量的影响。结果表明:(1)低于100 mg/L的氯霉素对小球藻和金藻8701叶绿素a含量影响较小(P>0.05);氯霉素浓度大于50 mg/L时,小新月菱形藻叶绿素a含量显著降低(P<0.05)。(2)不同浓度的G418均明显抑制三种微藻叶绿素a含量的增加。(3)低于100 mg/L的青霉素能够促进三种藻叶绿素a的增加,相对增长率随青霉素浓度的升高而逐渐下降;三种藻对青霉素的敏感性存在差异。试验结果可为微藻无菌培养体系的建立提供参考。