儿茶酚胁迫下铜绿微囊藻的生理生化响应

根据儿茶酚抑藻的剂量效应关系,研究了铜绿微囊藻在不同的投加剂量下,藻细胞的生理变化特征,采用扫描电子显微镜(SEM)观察细胞形貌,并测定了超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、胞内磷酸酶(AKP)和还原型谷胱甘肽酶(GSH)活性以及还原型抗坏血酸(AsA)和丙二醛(MDA)浓度的变化。结果显示,儿茶酚使细胞表面出现凹陷或者孔洞,致使丙二醛(MDA)相对浓度增加,生物膜受损明显。儿茶酚剂量≤EC50(0.80 mg/L)时,SOD、CAT和AKP酶活性在1~2 d增加,随后下降,接近于对照样。儿茶酚剂量≥EC60(1.20 mg/L)时,SOD、CAT和AKP酶活性被显著抑制,直到完全低于对照样水平。GSH和AsA也表现出类似规律。     

Identification and characterization of coagulation inhibitor proteins derived from cyanobacterium Microcystis aeruginosa

The excess growth of cyanobacteria in semi enclosed water areas caused by eutrophication brings about coagulation inhibition in drinking water treatment processes. In this study, coagulation inhibitor proteins produced by Microcystis aeruginosa, a major cyanobacterium in algal bloom, were acquired by a phage display technique, an aluminum-immobilized affinity chromatography and a protein expression technique using Escherichia coli cells. Two cyanobacterial peptides with a high ratio of metallophilic amino acids were recovered, which were a part of homologues of a thiol oxidase enzyme Ero1p and a trans-acting repressor ArsR. It was also shown that the homologue of ArsR exhibited a stronger inhibitory effect on the coagulation of kaolin suspension with polyaluminum chloride than the control proteins. This is the first report to identify a cyanobacterial cell component to inhibit coagulation. The compositions of polar amino acids were critical to explain the strength of coagulation inhibition potential. Polar proteins from cyanobacteria could collectively consume coagulants or stabilize clay particles, which would be plausible explanations for causing coagulation inhibition. Meanwhile, results from the kaolin coagulation tests using the control proteins implied that the neutralization of positive charges of coagulant constituents by simple electrostatic interactions might not be the key mechanism on the protein-induced coagulation inhibition.     

铜绿微囊藻生长条件优化模拟研究

采用Plackett Burman实验设计与响应曲面法(RSM)相结合,研究了环境因子对铜绿微囊藻生长的影响。首先采用Plackett-Burman设计对影响藻细胞生长的环境因子进行筛选,结果表明:温度、NaNO3、K2HPO4、Ferric.Citrate对铜绿微囊藻的生长有重要影响,各因子影响值大小为E温度EK2HPO4ENaNO3EFerric.Citrate。在此基础上,用Box-Behnken设计对铜绿微囊藻的生长条件进一步优化,通过对藻细胞最大现存量与各考察因子之间的输入响应关系进行分析,结果表明:铜绿微囊藻的最佳生长条件是:温度为29℃,NaNO3为1.059 mmol/L,K2HPO4为0.057 mmol/L,Ferric.Citrate为0.024mmol/L,此时藻细胞的最大现存量为10.8293×105cells/mL。     

The changes of nitric oxide production during the growth of Microcystis aerugrinosa

This study characterized the changes of nitric oxide (NO) production during the growth of Microcystis aerugrinosa, a cyanobacterium which usually cause cyanobacterial blooms. Results showed a drastic NO release accompanying with cell density and Chl-a content sharp rises when M. aerugrinosa grew from fifth day to sixth day. Moreover, high N:P ratio accelerated the cyanobacterial growth and NO burst. Sodium nitroprusside, an exogenous NO donor, promoted M. aerugrinosa growth with the optimal concentration of 0.1 mg/L. Experiments by supplementing with sodium nitrite and l-arginine demonstrated NO production in M. aerugrinosa cells was mainly through nitrate reductase (NR) pathway while minorly through NO synthase pathway. All these data suggested M. aerugrinosa produced increasing NO during its growth mainly by NR pathway, during which NO positively regulated the growth of M. aerugrinosa.     

Isolation and identification of an anti-algal compound from Artemisia annua and mechanisms of inhibitory effect on algae

The goals of this work were to isolate and identify an anti-algal compound from extracts of Artemisia annua and study its mode of action on Microcystis aeruginosa. The anti-algal compound was isolated from the extracts using column chromatography and activity-guided fractionation methods. Artemisinin with strong anti-algal activity was identified by gas chromatography-mass spectrometry and ¹H Nuclear Magnetic Resonance. The EC₅₀ of artemisinin on M. aeruginosa was 3.2 mg L⁻¹. Artemisinin decreased the soluble protein content and increased the superoxide dismutase activity and ascorbic acid content of M. aeruginosa, but exerted no effect on soluble sugar content. The results suggested the mode of action of artemisinin on algae may primarily be the increasing level of reactive oxygen species in algae cells. The results of our research could aid in the development of new anti-algal substances and lead to further study of mechanisms of inhibitory effect on algae.     

不同质量浓度苦草对铜绿微囊藻生长及抗氧化酶系统的影响

采用苦草（Vallisneria spiralis Linn.）和铜绿微囊藻（Microcystis aeruginosa）共生培养的实验方法,通过追踪测定铜绿微囊藻的生物量、叶绿素a含量、丙二醛（MDA）含量、超氧化物歧化酶（SOD）和过氧化物酶（POD）活性,研究了不同质量浓度苦草对铜绿微囊藻生长及抗氧化酶系统的影响。结果表明,质量浓度大于10 g/L时,苦草对铜绿微囊藻有明显的抑制作用,表现为苦草质量浓度为10、20和40 g/L时,第15天对铜绿微囊藻的抑制率分别为63.3%、94.7%和99.8%,培养过程中,铜绿微囊藻的叶绿素a含量逐渐减少,而SOD、POD活性及MDA含量呈现先增加后逐渐降低的趋势,表明苦草释放的化感物质在经过一定时间积累后能够明显抑制铜绿微囊藻SOD和POD的活性,引起细胞的氧化损伤,促进叶绿素的分解,从而导致藻类死亡,这是苦草抑制铜绿微囊藻生长的原因之一。     

Behavioural and biochemical responses of two marine invertebrates Scrobicularia plana and Hediste diversicolor to copper oxide nanoparticles

Engineered nano-sized Cu oxide particles are extensively used in diverse applications. Because aquatic environments are the ultimate “sink” for all contaminants, it is expected that nanoparticles (NP) will follow the same fate. In this study, two marine invertebrates Scrobicularia plana and Hediste diversicolor were chosen as ecotoxicological models. The aim was to evaluate behavioural (burrowing kinetics, feeding rate) and biochemical (biomarkers) responses of S. plana and H. diversicolor exposed in the laboratory to Cu (10 μg L⁻¹) added in natural seawater either in the form of engineered nanoparticles (NPs) of CuO or as dissolved Cu in 2% HNO₃. Exposure was characterized by considering (i) the physico-chemical fate of NP (ii) the fraction of labile Cu in experimental media and (iii) Cu bioaccumulation. Results showed high aggregation of CuO NPs in seawater and no additional bioavailable Cu concentrations. Behavioural impairments were observed in S. plana exposed to CuO NPs or soluble Cu whereas in H. diversicolor, only the exposure to soluble Cu led to a burrowing decrease. No obvious neurotoxicity effects were revealed since in both species, no changes in cholinesterasic activity occurred in response to both forms of Cu exposure. Biomarkers of oxidative-stress catalase and glutathione-S-transferase were enhanced in both species whereas superoxide dismutase was increased only in S. plana exposed to CuO NPs. Metallothionein-like protein was increased in bivalves exposed to both forms of Cu. Since, no detectable release of soluble Cu from CuO NPs occurred during the time of experiment, ecotoxicity effects seem to be related to CuO NPs themselves.     

稻秆对铜绿微囊藻抑制作用的研究

研究了稻秆浸出液对铜绿微囊藻生长的抑制作用.研究表明,稻秆浸出液中含有的化学物质抑制了铜绿微囊藻的生长繁殖.稻秆浸泡时间的延长,有利于提高浸出液的抑藻效果,但浸泡时间过长则无益于抑藻效果提高,浸泡时间为15 d时,抑制率能高达75.99%.稻秆浸出液投加量达7.5%以上具有显著的抑藻效果.不同部位的水稻秸秆浸出液对铜绿...     

Mechanisms of photosynthetic inactivation on growth suppression of Microcystis aeruginosa under UV-C stress

This study aims to investigate the effects of UV-C irradiation on photosynthetic processes of Microcystis aeruginosa to unravel the mechanism(s) involved in how and in what ways UV-C mediates growth suppression and cellular recovery. Changes in the concentration of photosynthetic pigments, photochemical efficiency, PS II core protein (D1) content, and the coding genes expressions were measured. The results indicate that UV-C doses at 20–200 mJ cm⁻² lead to rapid reduction in gene expression of both psbA (for D1) and cpc (for phycocyanin), but the suppression was short term and recoverable within 3 d of post-UV incubation. Conversely, UV-C doses at ?50 mJ cm⁻² could induce marked decline in photochemical efficiency (represented by the optimal PS II quantum yield, F_V/F_M, and the effective PS II quantum yield, Y) as well as decreases in D1 content and water soluble pigments (phycoerythrins, phycocyanins, allophycocyanins) in M. aeruginosa during the post UV-C incubation period. The results suggest that interruption of both the light energy harvesting apparatus (especially the water soluble pigments) and the photochemical process mainly accounted for the growth suppression effect in UV-C irradiated M. aeruginosa.     

Hematological, biochemical and ionoregulatory responses of Indian major carp Catla catla during chronic sublethal exposure to inorganic arsenic

In the present study the acute toxicity of arsenic trioxide in fingerlings of Catla catla an Indian major carp was evaluated with renewal bioassay method. The median lethal concentration of arsenic trioxide to the fish C. catla for 96 h was found to be 20.41 ppm (with 95% confidence limits). From this a non-lethal dose of (2.041 ppm; 1/10th of LC 50 96 value) was selected and fingerlings were exposed to 35 d and hematological, biochemical and ionoregulatory responses were studied at days 7, 14, 21, 28 and 35. Arsenic trioxide produced a significant increase in hemoglobin, hematocrit, WBC count, plasma GPT levels and reduction in RBC count, plasma sodium, chloride, potassium, glucose, protein, GOT, LDH levels as compared to the control group. Gill Na⁺/K⁺-ATPase activity was influenced by arsenic trioxide exposure. A biphasic response was noted in the value of MCH and MCV. However the MCHC level was not altered in arsenic trioxide treated fish throughout the study period. Results of the present investigation suggest that arsenic trioxide affects the hematological, biochemical and ionoregulatory parameters of fish and alterations of these parameters can be useful in environmental biomonitoring of arsenic contamination.     

聚铝及其加载粘土矿物高效絮凝沉降铜绿微囊藻的研究

考察了4种聚铝絮凝剂絮凝沉降铜绿微囊藻的效果,发现编号为HPACS02的聚铝絮凝剂除藻效果最好,并进一步研究得知HPACS02在pH 7.5时絮凝除藻的效率最高.在采用HPACS02作为絮凝剂时,如果少量投加滑石、海泡石或高岭土等粘土矿物会对铜绿微囊藻的絮凝沉降产生增效作用,其中滑石的助凝效果最好.     

聚铝及其加载粘土矿物高效絮凝沉降铜绿微囊藻的研究

考察了4种聚铝絮凝剂絮凝沉降铜绿微囊藻的效果，发现编号为HPACS02的聚铝絮凝剂除藻效果最好，并进一步研究得知HPACS02在pH7．5时絮凝除藻的效率最高。在采用HPACS02作为絮凝剂时，如果少量投加滑石、海泡石或高岭土等粘土矿物会对铜绿微囊藻的絮凝沉降产生增效作用，其中滑石的助凝效果最好。     

Inter- and intra-specific responses to elevated ozone and chamber climate in northern birches

We studied the responses of micropropagated, northern provenances of downy, mountain and silver birches to elevated ozone (O₃) and changing climate using open-top chambers (OTCs). Contrary to our hypothesis, northern birches were sensitive to O₃, i.e. O₃ levels of 31-36 ppb reduced the leaf and root biomasses by −10%, whereas wood biomass was affected to a lesser extent. The warmer and drier OTC climate enhanced growth in general, though there were differences among the species and clones, e.g. in bud burst and biomass production. Inter- and intra-specific responses to O₃ and changing climate relate to traits such as allocation patterns between the above- and belowground parts (i.e. root/shoot ratio), which further relate to nutrient and water economy. Our experiments may have mimicked future conditions quite well, but only long-term field studies can yield the information needed to forecast responses at both tree and ecosystem levels.     

不同水温下鲴鱼对铜绿微囊藻的控制作用及对水质的影响

在室内受控模拟条件下开展实验,研究了在19、23、27、31、35℃5个水温梯度下鲴鱼对铜绿微囊藻和水质的影响。研究结果表明,在不同水温下,鲴鱼对铜绿微囊藻具有较强的控制作用,实验结束时铜绿微囊藻密度减少至初始密度的18%~30%,摄食率和消化率分别为6.83×104~8.32×104cells/(g·d)、93%~98%;叶绿素a的去除率为68%~88%;实验组TP、TN去除率分别为22%~25%、20%~38%,对照组的分别为80%~94%、28%~40%。对照组NH+4-N浓度变化很小(0.071~0.073 mg/L),而实验组氨氮浓度显著增大(2.222~3.645 mg/L),分别为初始值的31、34、42、51和46倍。     

水华束丝藻与铜绿微囊藻净化水体氮磷及其脂质积累过程比较

选取水华束丝藻与铜绿微囊藻作为典型水华蓝藻,探索水华蓝藻净化水体及合成生物柴油的潜力.采用氮磷浓度为NH4+-N 7.5 mg/L,PO43--P 1.0 mg/L的模拟污水,重点考察了在光照强度与温度不同的条件下2种微藻对氮磷的去除率,并利用尼罗红荧光染色法测定了微藻中的中性脂含量.结果表明,2种藻对PO43--P的去除率均较高,最高达到98%,对NH4+-N的去除率稍低,最高为52%.水华束丝藻及铜绿微囊藻均具有较强积累中性脂的能力,适宜的温度与光照条件有利于微藻对氮磷的去除及细胞中脂类的积累.2种藻均在光照强度为12 000 lx时中性脂含量更高.水华束丝藻在温度为18~22℃条件下,生长更优,氮磷净化效果较好,中性脂积累能力更强.而铜绿微囊藻在温度为22~26℃时表现更好.     

Antioxidant and metabolism responses to polyphenol stress in cyanobacterium Microcystis aeruginosa

Three common polyphenol compounds Gallic Acid (GA), Pyrogallic Acid (PA) and Catechol (CA) are known to have allelochemical-exhibiting inhibitory effects on the growth of the cyanobacterium Microcystis aeruginosa (M. aeruginosa). Metabolism and antioxidant responses in M. aeruginosa were investigated to elucidate the mechanism by which the three polyphenols inhibit algal growth. The inhibition effects of polyphenols were in the order of CA > PA > GA. The GA and CA exposures increased protein contents, superoxide dismutase (SOD) activity, catalase (CAT) activity and soluble sugar, especially for exposure to GA of 25 mg L^?1. Soluble sugar content increased significantly especially when exposed to CA for 72 h. When exposed to PA, protein content, and SOD and CAT activities initially increased but over longer treatment time the activities decreased, in contrast to sugar content. Our results suggest that PA exposure for longer periods of time may inhibit catabolism action, while CA exposure could induce more oxide stress than GA or PA. The overall study showed that polyphenol-induced oxidative damage might be responsible for polyphenol inhibition on M. aeruginosa growth. The increases in cellular antioxidant enzymes and soluble sugar may have been to counteract the oxidative stress.     

碳源对铜绿微囊藻生理特性及微囊藻毒素产率的影响

为研究水体中不同碳源对铜绿微囊藻生理特性的影响,以Na2CO3与葡萄糖分别作为铜绿微囊藻生长的无机碳源与有机碳源,将铜绿微囊藻于光照下进行培养,并对其一系列的生理特性与微囊藻毒素产率进行检测。实验结果表明,同等碳浓度下,有机碳源更能促进铜绿微囊藻的生长,经过30 d的培养,铜绿微囊藻在有机碳源中的产量为187.55 g,比其在无机碳源中的产量提高了6.06%;微囊藻毒素在有机碳源中的产量为969.00μg/g,而在无机碳源中的产量却升高至1 193.60μg/g。参与藻毒素合成的3种氨基酸在无机碳源中的浓度要比有机碳源中的浓度高,但是其余几种氨基酸的含量与之情况相反。而有机碳源培养的铜绿微囊藻总可溶性蛋白含量为387.00μg/g,比无机碳源培养的铜绿微囊藻的蛋白含量提高了93.60%。     

Short and long-term effects of clofibric acid and diclofenac on certain biochemical and ionoregulatory responses in an Indian major carp, Cirrhinus mrigala

Extensive use of pharmaceuticals in human and veterinary medicine and aquaculture practices pose a serious threat to aquatic organisms. In the present investigation, Cirrhinus mrigala an Indian major carp was exposed to different concentrations of clofibric acid (CA) and diclofenac (DCF) and certain biochemical and ionoregulatory responses were assessed under short and long term exposures. During short-term (96 h) exposure period, plasma glucose and sodium (Na⁺) levels were increased at all concentrations (1, 10 and 100 μg L⁻¹) of CA and DCF treated fish. Plasma protein and chloride (Cl⁻) levels were found to be decreased at all concentrations of CA and DCF exposed fish comparatively to control groups. Meanwhile an increase in plasma potassium (K⁺) level was noted in fish exposed to CA treatments alone and in DCF treatments it was decreased. In long-term exposure (35 d), plasma Na⁺ and Cl⁻ levels were found to be significantly increased at all concentration of CA and DCF. However, a biphasic trend was observed in plasma glucose, protein and K⁺ levels. In both the treatments, a significant (P < 0.01 and P < 0.05) changes were observed in all parameters measured in fish exposed to different concentrations of CA and DCF. The results of the present investigation indicate that both the drugs caused significant changes in biochemical and ionoregulatory responses of fish at all concentrations. The alterations of these parameters can be useful in monitoring of pharmaceutical residues present in aquatic environment.     

Growth and photosynthetic responses to copper in wild grapevine

The present study evaluates the tolerance and accumulation potential of Vitis vinifera ssp. sylvestris under moderate and high external Cu levels. A greenhouse experiment was conducted in order to investigate the effects of a range of external Cu concentrations (0–23 mmol L⁻¹) on growth and photosynthetic performance by measuring gas exchange, chlorophyll fluorescence parameters and photosynthetic pigments. We also measured the total copper, nitrogen, phosphorus, sulphur, calcium, magnesium, iron, potassium and sodium concentrations in the plant tissues. All the experimental plants survived even with external Cu concentrations as high as 23 mmol L⁻¹ (1500 mg Cu L⁻¹), although the excess of metal resulted in a biomass reduction of 35%. The effects of Cu on growth were linked to a reduction in net photosynthesis, which may be related to the effect of the high concentration of the metal on photosynthetic electron transport. V. vinifera ssp. sylvestris survived with leaf Cu concentrations as high as 80 mg kg⁻¹ DW and growth parameters were unaffected by leaf tissue concentrations of 35 mg Cu kg⁻¹ DW. The results of our study indicate that plants of V. vinifera ssp. sylvestris from the studied population are more tolerant to Cu than the commercial varieties of grapevine that have been studied in the literature, and could constitute a basis for the genetic improvement of Cu tolerance in grapevine.