Salicylic acid and heat acclimation pretreatment protects Laminaria japonica sporophyte (Phaeophyceae) from heat stress

Possible mediatory roles of heat acclimation and salicylic acid in protecting the sporophyte of marine macroalga Laminaria japonica (Phaeophyceae) from heat stress were studied. Heat stress resulted in oxidative injury in the kelp blades. Under heat stress significant accumulation of hydrogen peroxide (H₂O₂) and malonaldehyde (MDA), a membrane lipid peroxidation product, and a drastic decrease in chlorophyll a content were recorded. Activity of the enzymatic antioxidant system was drastically affected by heat stress. The activity of superoxide dismutase (SOD) was significantly increased while peroxidase (POD), catalase (CAT) and glutathione peroxidase (GPX) were greatly inhibited and, simultaneously, phenylalanine ammonia-lyase was activated while polyphenol oxidase (PPO) was inhibited. Both heat acclimation pretreatment and exogenous application of salicylic acid alleviated oxidative damage in kelp blades. Blades receiving heat acclimation pretreatment and exogenous salicylic acid prior to heat stress exhibited a reduced increase in H₂O₂ and MDA content, and a lower reduction in chlorophyll a content. Pretreatment with heat acclimation and salicylic acid elevated activities of SOD, POD, CAT, GPX and PPO. Considering these results collectively, we speculate that the inhibition of antioxidant enzymes is a possible cause of the heat-stress-induced oxidative stress in L. japonica, and enhanced thermotolerance may be associated, at least in part, with the elevated activity of the enzymatic antioxidant system.     

Studies on the effects on growth and antioxidant responses of two marine microalgal species to uniconazole

Uniconazole, as a plant growth retardant, can enhance stress tolerance in plants, possibly because of improved antioxidation defense mechanisms with higher activities of superoxide dismutase(SOD) and peroxidase(POD) enzymes that retard lipid peroxidation and membrane deterioration. These years much attention has been focused on the responses of antioxidant system in plants to uniconazole stress, but such studies on aquatic organism are very few. Moreover, no information is available on growth and antioxidant response in marine microalgae to uniconazole. In this paper, the growth and antioxidant responses of two marine microalgal species, Platymonas helgolandica and Pavlova viridis, at six uniconazole concentrations(0-15 mg L-1) were investigated. The results demonstrated that 3 mg L-1 uniconazole could increase significantly chlorophyll a and carbohydrate contents of P. helgolandica(P 0.05). Higher concentrations(≥12 mg L-1) of uniconazole could inhibit significantly the growth, dry weight, chlorophyll-a and carbohydrate contents of P. helgolandica and P. viridis(P 0.05). Uniconazole caused a significant increase in lipid peroxidation production(MDA) at higher concentrations(≥ 9 mg L-1). The activities of antioxidant enzymes, superoxide dismutase(SOD) and catalase(CAT) were enhanced remarkably at low concentrations of uniconazole. However, significant reduction of SOD and CAT activities was observed at higher concentrations of uniconazole.     

Antioxidation and ATPase activity in the gill of mud crab Scylla serrata under cold stress

Mud crab (Scylla serrata) is an important commercial crustacean in China. An experiment was designed to study the effect of cold stress on S. serrata. After a one-week adaptation at 28oC, the temperature is suddenly reduced to 4oC. The crabs were sampled every 2 h for 10 h and dissected immediately to measure the enzyme activity. The crabs at room temperature (28oC) were used as the control group. The activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX), the content of malondialdehyde (MDA) and the activity of 4 ATPases (Na , K -ATPase; Mg2 -ATPase; Ca2 -ATPase; Ca2 , Mg2 -ATPase) were measured biochemically. In contrast to the control group, the SOD activity increased significantly from 2 to 6 h after the cold stress, and then decreased. The CAT and GPX activities increased in 2 h, and then decreased gradually. The content of MDA increased gradually in 4 h. The activity of Na , K -ATPase decreased in 2 h, increased up to the top value at Hour 6, then decreased again. The activities of Mg2 -ATPase, Ca2 -ATPase and Ca2 , Mg2 -ATPase increased significantly in 6 h, insignificantly in any other hours. Under cold stress, the activity of antioxidative enzymes in S. serrata was reduced at first then stabilized, ROS-scavenging weakened, and MDA accumulated gradually in the gill after 6 h. The activity of the 4 ATPases in the crab decreased after 6 h, suggesting that the ability to regulate ion concentration has been paralyzed. Therefore, the maximum period to sustain healthy meat in the crab under cold stress is 6 hours.     

Effect of dietary supplementation of vitamin C on growth,reactive oxygen species,and antioxidant enzyme activity of Apostichopus japonicus (Selenka) juveniles exposed to nitrite

Different amounts of vitamin C were added to diets fed to juveniles(2.5 ± 0.15 g) of sea cucumber Apostichopus japonic u s(Selenka) in an attempt to reduce the stress response of specimens exposed to nitrite stress. A commercial feed was used as the control diet and three experimental diets were made by supplementing 1 000, 1 500, or 2 000 mg vitamin C/kg diet to control diet separately in a 45-day experiment. Sea cucumbers were exposed to three different levels(0.5, 1.0, and 1.5 mg/L) of nitrite stress for 4, 8, and 12 h at four time intervals(0, 15, 30, and 45 d). Growth of the animals was recorded during the experiment. Reactive oxygen species(ROS)(i.e. hydroxyl free radical(-OH), malondialdehyde(MDA) and total antioxidant capacity(T-AOC)) and antioxidant enzyme activities(i.e., superoxide dismutase(SOD) and catalase(CAT)) were measured. Response surface methodology(RSM) was used to analyze the effect of multiple factors on ROS indices and enzyme activities. Weight gain(WG) and special growth rate(SGR) of vitamin C supplementation groups were significantly higher than those of control group( P 0.05). The levels of-OH and MDA increased under exposure time extending and nitrite concentration increasing, whereas T-AOC level decreased. SOD and CAT activities increased at 4 h and 8 h and decreased at 12 h. During the days in which the animal consumed experimental diets, the levels of-OH and MDA decreased and that of T-AOC increased. This result suggests that diets containing vitamin C could reduce the nitrite stress response in the animals and increase their antioxidant capacity. The multifactor regression equation of growth performance, ROS indices, and duration of feeding results suggest that vitamin C supplementation of 1 400–2 000 mg/kg diet for 29–35 days could reduce effectively the effects of nitrite exposure.     

Stress Responses to Rapid Temperature Changes of the Juvenile Sea Cucumber （Apostichopusjaponicus Selenka）

Activities of hexokinase （HK）, pyruvate kinase （PK）, superoxide dismutase （SOD） and catalase （CAT） and Hsp70 level were measured to evaluate the response of the commercially important sea cucumber （Apostichopus japonicus Selenka） to rapid temperature changes in laboratory. Animals were subjected to a higher temperature （from 10 to 20℃） （Tinc treatment） or to a lower temperature （from 20 to 10℃） （Tddec treatment） for 72 h. At 1, 3, 12, 24, 72 h of exposure, animals were removed and prepared for further analysis. Results showed that the effect of acute temperature changes on enzyme activities was significant. In Tinc treatment, activities of SOD and CAT increased immediately. The significant enhancement of SOD and CAT activities suggested that oxidative stress increases significantly when ambient temperature increasing from 10 to 20℃. The up-regulation of Flsp70 in Tinc and Tdec treatments indicated that Hsp70 was a bioindicator of thermal stress in the sea cucumber, and the expression pattern depended on the thermal treatment.     

Antiaging activity of low molecular weight peptide from Paphia undulate

Low molecular weight peptide (LMWP) was prepared from clam Paphia undulate and its antiaging effect on D-galactose-induced acute aging in rats, aged Kunming mice, ultraviolet-exposed rats, and thermally injured rats was investigated. P. undulate flesh was homogenized and digested using papain under optimal conditions, then subjected to Sephadex G-25 chromatography to isolate the LMWP. Administration of LMWP significantly reversed D-galactose-induced oxidative stress by increasing the activities of glutathione peroxidase (GPx) and catalase (CAT), and by decreasing the level of malondialdehyde (MDA). This process was accompanied by increased collagen synthesis. The LMWP prevented photoaging and promoted dermis recovery and remission of elastic fiber hyperplasia. Furthermore, treatment with the LMWP helped to regenerate elastic fibers and the collagen network, increased superoxide dismutase (SOD) in the serum and significantly decreased MDA. Thermal scald-induced inflammation and edema were also relieved by the LWMP, while wound healing in skin was promoted. These results suggest that the LMWP from P. undulate could serve as a new antiaging substance in cosmetics.     

Variations of antioxidant enzyme activity and malondialdehyde content in nemertean Cephalothrix hongkongiensis after exposure to heavy metals

The antioxidant enzyme activity and malondialdehyde (MDA) content of Cephalothrix hongkongiensis were studied to assess variations in the biochemical/physiological parameters of nemerteans under heavy metal stress. Worms were exposed to copper, zinc and cadmium solutions at different concentrations, and the activity of three antioxidant enzymes, catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPX), and MDA content were measured. The results show that the activity of each enzyme changed immediately after exposure to heavy metals. CAT was invariably inhibited throughout the experimental period, while the SOD activity was significantly elevated by exposure to Cu²⁺ for 48 h, but then decreased. SOD was inhibited by Zn²⁺during the first 12 h of exposure, but activated when exposed for longer periods. Under Cd²⁺ stress, SOD activity decreased within 72 h. GPX activity varied greatly, being significantly increased by both Cu²⁺ and Zn²⁺, but significantly inhibited by Cd²⁺ in the first 12–24 h after exposure. MDA content increased on Cu²⁺ exposure, but normally decreased on Zn²⁺ exposure. MDA content followed an increase-decrease-increase pattern under Cd²⁺ stress. In conclusion, the antioxidant system of this nemertean is sensitive to heavy metals, and its CAT activity may be a potential biomarker for monitoring heavy metal levels in the environment.     

Effects of Environmental Cadmium on Cadmium Accumulation,Oxidative Response,and Microelements Regulation in the Liver and Kidney of Hexagrammos otakii

The effects of cadmium(Cd)on metal accumulation,microelements contents,and antioxidant responses in Hexagram-mos otakii were studied.The fish were exposed to 0.2,2.5,or 10μg L?1 Cd for 12 or 24 days.Then,the concentration of Cd and mi-croelements(Ca,Fe,Zn,and Se)were determined in the liver and kidney.Moreover,the activities of antioxidant enzymes including superoxide dismutase(SOD),catalase(CAT),and glutathione-S-transferase(GST),and the content of malondialdehyde(MDA)in the liver and kidney were also measured.A continuous accumulation of Cd was observed throughout the experimental period.Cd accumulation was higher in liver than that in the kidney,while Ca and Fe contents were lower in liver than those in the kidney.Cd exposure resulted in a decrease of Ca and Fe concentrations in the kidney,while there was no effect on the liver.Zn and Se remained unaffected with exposure to Cd.Cd exposure induced severe oxidative stress in H.otakii,as indicated by significant induction of the activities of SOD,CAT,and GST,and a simultaneous increase of MDA content.These data show that antioxidant enzymes and mi-croelements contents can be used as potential biomarkers to monitor environmental health in fish.     

Effects of chilled storage and cryopreservation on sperm characteristics,antioxidant enzyme activities,and lipid peroxidation in Pacific cod <Emphasis Type="Italic">Gadus microcephalus</Emphasis>

The present study evaluated the effects of chilled storage and cryopreservation on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod Gadus macrocephalus. Sperm motility and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (Gr), and lipid peroxidation (measured via malondialdehyde (MDA) content) were determined after the milt was stored at 4°C for 12 h, cryopreserved without cryoprotectant in 12% propylene glycol (PG), cryopreserved in 12% PG+0.1 mol/L trehalose, or cryopreserved in 12% PG spermatozoa but centrifuged to decant the supernatant prior to cryopreservation (only sperm cells were cryopreserved). After chilled storage or cryopreservation, the SOD, CAT and GPx activities were reduced in sperm cells and increased in seminal plasma in almost all treatments; sperm motility parameters were also decreased. However, the addition of trehalose into the cryoprotectant could significantly improve the postthaw sperm quality as revealed by the sperm average path velocity. This improvement might be attributed to the function of trehalose in scavenging reactive oxygen species. Chilled storage and cryopreservation had significant effects on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod.     

Response of Yesso scallop Patinopecten yessoensis to acute temperature challenge: physiological and biochemical parameters

Water temperature is generally considered to be a major factor affecting the physiological and biochemical activities of marine bivalves. Here, the physiological and biochemical responses of Yesso scallop, Patinopecten yessoensis, to acute water temperature changes in summer were studied. Scallops were transferred directly to a lower temperature (T_dec treatment) (from 23°C to 15°C) or to a higher temperature (T_dec treatment) (from 15°C to 23°C) for 72 h, respectively. Results showed that the oxygen consumption and ammonia-N excretion rates of P. yessoensis decreased significantly in the T_dec treatment but increased dramatically at 6 h in the T_dec treatment (P <0.05). In the T dec treatment, hepatopancreas antioxidant enzyme activities, superoxide dismutase (SOD) and catalase (CAT) activities, increased substantially within 72 h (P <0.05). However, a significant decrease in CAT activity was found at 12 h in the T_dec treatment (P <0.01). A significant enhancement of acid phosphatase (ACP) activity and malondialdehyde (MDA) content was detected when scallops were acutely exposed to a temperature of 15°C. The levels of Cu/Zn-SOD gene expression in their gills up-regulated significantly in response to acute temperature changes (P <0.01). These data suggest that acute temperature change affects physiological and biochemical functions, and improve our knowledge of P. yessoensis under conditions of thermal stress.

     

干旱胁迫下植物抗逆性大尺度遥感监测方法

目前干旱与植被关系的研究主要集中于气候因子与植被时空变化的相关性分析以及植被对气候变化的响应,能够适用于大尺度的植物抗逆性监测方法还较为欠缺。本文基于归一化植被指数（NDVI）、总初级生产力（GPP）、修正花青素含量指数（mACI）、短波红外水分胁迫指数（SIWSI）监测干旱胁迫下的植被变化,综合考虑植物抗逆过程,建立滞后时间、抗逆时差、响应程度与恢复能力4个植物抗逆性监测指标,构建了一种能够适用于大尺度的植物抗逆性综合监测方法。利用各省份作物抗逆性综合评分与绝收比例进行相关性分析,两者呈显著负相关。利用该方法对干旱胁迫下我国不同类型植被的抗逆性进行评估,结果表明：(1)从全国整体水平来看,不同季节植物抗逆性差异较大,其中夏季植物抗逆性最弱,冬季最强。我国植物抗逆性空间异质性显著,春季植物抗逆性综合评分低于70分的区域主要位于山西、陕西北部,综合评分高于90分的区域主要集中在内蒙古东北部以及云南的南部地区;(2)不同类型植被的抗逆性有明显差异,夏季落叶针叶林抗逆性最强,类内差异最小,春秋两季草地抗逆性最强但抗逆性类内差异最大。本文提出的植物抗逆性综合监测方法有助于探索干旱胁迫下植物抗逆...     

Use of antioxidant enzymes of clam <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis> as biomarker to polycyclic aromatic hydrocarbon pollution

The typical organic pollutant polycyclic aromatic hydrocarbon (PAH) anthracene was selected as a contaminant to investigate its effects on the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in the clam Ruditapes philippinarum. The results show that SOD, CAT and GSH-Px had diff erent induction and inhibition reactions to anthracene stress, and that three diff erent organs in R. philippinarum (visceral mass, muscle tissue and mantle) had diff erent sensitivities to anthracene stress. This study suggest that SOD activities of the visceral mass, CAT activitities of the mantle and the visceral mass, and GSH-Px activity of the muscle tissue could be used as sensitive indicators of anthracene stress in R. philippinarum.     

Effect of polysaccharide from Enteromorpha prolifera on maize seedlings under NaCl stress

In this study,a polysaccharide from Enteromorpha prolifera(EP)was extracted and its effect on maize seedlings under NaCl stress was investigated.Firstly,the components and structure of the EP were determined.We found that EP is a sulfated polysaccharide of high-molecular weight(Mw,1 840 KDa)heteropoly saccharides and the main mono saccharide is rhamno se.The poly saccharide was applied to explore its effect on the growth of maize seedlings and its defense response under a salt stress.The results show that EP could promote the growth of maize seedlings under the salt stress.In addition,EP was shown able to significantly regulate membrane permeability and adjustment of osmotic substances such as soluble protein,soluble sugar,and proline,antioxidant enzymes containing superoxide dismutase,catalase,peroxidase,and ascorbate peroxidase.Therefore,EP is an effective salt-resistant substance for the growth of maize seedlings under NaCl stress.     

CO2协同低温有水对卵形鲳鲹麻醉保活效果的影响

【目的】探究二氧化碳CO₂麻醉协同低温对卵形鲳鲹有水保活的影响,延长卵形鲳鲹(Trachinotus ovatus)的保活时间。【方法】研究保活过程中CO₂质量浓度、麻醉时间、麻醉温度和鱼质量水比对卵形鲳鲹有水保活时长的影响;在最佳麻醉保活条件下,测定其血清生化指标、肌肉及肝脏氧化应激指标及代谢指标。【结果】卵形鲳鲹在16℃、100 mg/L CO₂水溶液中麻醉3 min其麻醉效果最佳。在15℃低温下进行有水保活,鱼水质量比为2∶1,存活时间最长,为(359±2)min,此条件下复苏率达100%。经过CO₂麻醉协同低温有水保活后的卵形鲳鲹血清谷草转氨酶(GOT)活性显著升高(P<0.05)、尿素氮(BUN)和甘油三酯(TG)含量上升,血糖含量下降。肌肉中的乳酸脱氢酶(LDH)活性显著上升(P<0.05),乳酸(LD)和糖原含量上升,过氧化氢酶(CAT)活性和丙二醛(MDA)含量下降;肝脏中的LDH活性显著上升(P<0.05)、CAT活性和LD的含量上升,MDA和糖原的含量下降。【结论】CO₂协同低温有水对卵形鲳鲹有较好的麻醉效果,有助于保活运输。在保活过程中卵形鲳鲹通过提升能量代谢水平和抗氧化能力应答环境应激,在一定程度上影响卵形鲳鲹保活过程的存活时间。     

Mercury (Hg2+) effect on enzyme activities and hepatopancreas histostructures of juvenile Chinese mitten crab Eriocheir sinensis

We studied the effects of mercury (Hg²⁺) on antioxidant and digestive enzyme activities in terms of LC₅₀ value and on hepatopancreas histostructures of juvenile Chinese mitten crabs Eriocheir sinensis in 40-day exposure to various concentrations of Hg²⁺ (0, 0.01, 0.05, 0.10, 0.20, and 0.30 mg/L). The results show that the activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) significantly increased in the concentrations of 0.01 and 0.05 mg/L, while that of enzyme decreased in 0.10, 0.20 and 0.30 mg/L treatments. Meanwhile, Hg²⁺ disrupted the histostructures of the hepatopancreas, causing decreases in activities of pepsin, tryptase, amylase, and cellulose, which are synthesized in the hepatopancreas. Moreover, as the Hg²⁺ concentration increased, the survival rate of the crabs decreased, worst at 56.57% in 0.30 mg/L. Therefore, although crabs are able to tolerate low levels of mercury pollution, high levels lead to cellular injury and tissue damage in hepatopancreas, which then loses some of its vital physiological functions such as absorption, storage, and secretion.     

Antioxidant and isozyme features of two strains of Laminaria japonica （Phaeophyceae）

Healthy sporophytes of two gametophyte mutants of Laminaria japonica with different heat resistances: kelp 901 (901, with comparatively stronger heat-resistance) and Rongcheng No.1 (RC, sensi- tive to heat stress), were respectively collected during October to December 2002 from Yantai and Rong- cheng Sea Farm in the Shandong Peninsula of China. The contents of some biochemical materials and antioxidant capacity were analyzed under controlled laboratory conditions to identify if there is any rela- tion between the overall antioxidant capacity and the heat-resistance in L. japonica and to understand possible mechanism of heat-resistance. Results show that: (1) the overall antioxidant capacity in healthy sporophyte of 901, such as vitamin E, polyphenol, and ascorbic acid contents and the enzymatic activity of SOD, POD, CAT, Gpx, PPO, and PAL, were not always higher than that of RC under controlled labo- ratory conditions, and no significance (P>0.05) was shown in total antioxidant capacity (T-AOC) in 901 and RC. Result suggested that the difference in antioxidant capacity was not a decisive factor for different heat-resistances in L. japonica; (2) the simultaneous assay on isozymes was carried out using vertical polyacrylamide gel electrophoresis (PAGE). Considerable differences in peroxide (PRX), malate dehy- drogenase (MDH), malic enzyme (ME), polyphenol oxidase (PPO) and glutamate dehydrogenase (GDH) were obtained in 901 and RC from either the band number, relative mobility (Rf), or staining intensity, and ME could be used as an indicator to distinguish healthy sporophyte of 901 and RC under controlled labo- ratory conditions.     

Use of antioxidant enzymes of clam Ruditapes philippinarum as biomarker to polycyclic aromatic hydrocarbon pollution

The typical organic pollutant polycyclic aromatic hydrocarbon(PAH) anthracene was selected as a contaminant to investigate its eff ects on the activities of superoxide dismutase(SOD), catalase(CAT) and glutathione peroxidase(GSH-Px) in the clam Ruditapes philippinarum. The results show that SOD, CAT and GSH-Px had diff erent induction and inhibition reactions to anthracene stress, and that three diff erent organs in R. philippinarum(visceral mass, muscle tissue and mantle) had diff erent sensitivities to anthracene stress. This study suggest that SOD activities of the visceral mass, CAT activitities of the mantle and the visceral mass, and GSH-Px activity of the muscle tissue could be used as sensitive indicators of anthracene stress in R. philippinarum.     

Chitosan oligosaccharide-Ca complex accelerates the depuration of cadmium from Chlamys ferrari

This study investigated the effect of a chitosan oligosaccharide-Ca complex (COS-Ca) on the depuration of cadmium (Cd) from Chlamys ferrari. After exposure to 0.5 mg L-1 CdCl2 for 3 or 7 d, the scallops were treated by COS-Ca prior to determina-tion of Cd, calcium (Ca) and zinc (Zn) contents, Cd distribution in organs, malondialdehyde (MDA) content and antioxidant variables. Results showed that COS-Ca reduced Cd content in the viscera of the scallops, with highest Cd depuration rate (47%) observed on day 3. The COS-Ca concentration substantially affected Cd depuration, and the exposure to 8.75 mg L-1 COS-Ca led to significantly higher Cd depuration rate compared with those of lower COS-Ca concentrations (1.75, 3.5, 5.25, and 7.00 mg L-1). Distribution analysis of Cd in scallop organs indicated that COS-Ca significantly reduced Cd content in the kidney throughout the 5-d experiment, as well as in the gill during the early stage of Cd depuration. In addition, COS-Ca treatment decreased glutathione peroxidase (GSH-Px) activity and MDA content while increasing superoxide dismutase (SOD) and catalase (CAT) activities on different days. Our work suggested COS-Ca complex treatment as an effective method for acceleration of Cd depuration from Cd-contaminated bivalves.     

Effects of benzo（a）pyrene exposure on the antioxidant enzyme activity of scallop Chlamysfarreri

Scallop Chlamys farreri was exposed to different concentrations of benzo(a)pyrene (BaP) (0.5 μg/L, 1.0 μg/L, 10.0 μg/L and 50.0 μg/L) for 30 days in seawater. The 7-ethoxyresorufin O-deethylase (EROD) activity was significantly induced, and increased with the increasing BaP concentration. The glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), Glutathione peroxidase (GPx) activities increased in short time at low concentration of BaP, and was significantly depressed at high concentr...     

Antioxidative capacity and enzyme activity in Haematococcus pluvialis cells exposed to superoxide free radicals

The antioxidative capacity of astaxanthin and enzyme activity of reactive oxygen eliminating enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were studied in three cell types of Haematococcus pluvialis exposed to high concentrations of a superoxide anion radical (O₂⁻). The results show that defensive enzymes and astaxanthin-related mechanisms were both active in H. pluvialis during exposure to reactive oxygen species (ROS) such as O₂⁻. Astaxanthin reacted with ROS much faster than did the protective enzymes, and had the strongest antioxidative capacity to protect against lipid peroxidation. The defensive mechanisms varied significantly between the three cell types and were related to the level of astaxanthin that had accumulated in those cells. Astaxanthin-enriched red cells had the strongest antioxidative capacity, followed by brown cells, and astaxanthin-deficient green cells. Although there was no significant increase in expression of protective enzymes, the malondialdehyde (MDA) content in red cells was sustained at a low level because of the antioxidative effect of astaxanthin, which quenched O₂⁻ before the protective enzymes could act. In green cells, astaxanthin is very low or absent; therefore, scavenging of ROS is inevitably reliant on antioxidative enzymes. Accordingly, in green cells, these enzymes play the leading role in scavenging ROS, and the expression of these enzymes is rapidly increased to reduce excessive ROS. However, because ROS were constantly increased in this study, the enhance enzyme activity in the green cells was not able to repair the ROS damage, leading to elevated MDA content. Of the four defensive enzymes measured in astaxanthin-deficient green cells, SOD eliminates O₂⁻, POD eliminates H₂O₂, which is a by-product of SOD activity, and APX and CAT are then initiated to scavenge excessive ROS.