Toxicity and Accumulation of Selenite in Four Microalgae

The toxicity and bioaccumulation of selenite in four microalgae, Spirulina platensis,Dunaliella salina, DunalieUa bardawill and Phaeodactylum tricornutum cultured in the presence of selenite were investigated. Lower concentrations of selenite were generally nontoxic and frequently stimulated algal growth, while higher concentrations of selenite inhibited algal growth. Selenite was more toxic to D. salina and D. bardawill than to S. platensis and P. tricornutum. All algae cultured in selenite were able to incorporate Se to different degrees, which depended on algal species. The distributions of selenite among intracellular macromolecular compounds were different among algal species : most of the selenite was associated with proteins in S. platensis, D. salina and D. bardawill, while most of the selenite was associated with lipids in P. tricornutum, which reflected the physiological differences among the algae. These observations suggest that algae are able to accumulate selenite and bind it with intracell~ar macromolecular compounds when exposed to high concentration of selenite. This may represent a form of storage or detoxification of selenite by the algae.     

Recovery of gonadal development in tiger puffer Takifugu rubripes after exposure to 17β-estradiol during early life stages

The aim of the present study was to investigate the long-term effects of 17β-estradiol(E2) exposure on gonadal development in the tiger puffer( Takifugu rubripe s), which has a genetic sex determination system of male homogametic XY-XX. Tiger puffer larvae were exposed to 1, 10 and 100 μg/L E2 from 15 to 100 days post-hatch(dph) and then maintained in clean seawater until 400 dph. Changes in sex ratio, gonadal structure and gonadosomatic index(GSI) were monitored at 100, 160, 270 and 400 dph. Sex-associated single nucleotide polymorphism(SNP) markers were used to analyze the genetic sex of samples, except those at 100 dph. Exposure had a positive effect on the conversion of genetically male gonads into phenotypically female gonads at 100 dph. However, gonads from 60% of genetic XY males in the 1-μg/L E2 group and 100% in the 10-μg/L E2 group developed intersexual gonads at 160 dph; gonads of all genetic XY males in the two treatment groups reverted to testis by 270 dph. While 38%, 57% and 44% of gonads of XY fish in the 100-μg/L E2 group reverted to intersexual gonads at 160, 270 and 400 dph, respectively, none reverted to testis after E2 treatment. In addition, E2 exposure inhibited gonadal growth of both genetic sexes, as indicated by the clear dose-dependent decrease in GSI at 270 and 400 dph. The results showed that exposure to E2 during the early life stages of tiger puffer disrupted gonadal development, but that fish recovered after migration to clean seawater. The study suggests the potential use of tiger puffer as a valuable indicator species to evaluate the effects of environmental estrogens on marine fish, thereby protecting valuable fishery resources.     

EFFECT OF GERMANIUM DIOXIDE ON GROWTH OF SPIRULINA PLATENSIS

This study on the effect of different concentrations of germanium dioxide (GeO_2) on the specificgrowth rate (SGR), pigment contents, protein content and amino acid composition of Spirulina platensisshowed that Ge was not the essential element of this alga; that GeO_2 could speed up growth and raiseprotein content of S. platensis, and could possibly influence the photosynthesis system. The concentrationrange of GeO_2 beneficial to growth of S. platensis is from 5-100 mg/l. GeO_2 is proposed to be util-ized to remove contamination by Chlorella spp. usually occurring in the cultivation of Spirulina.     

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...     

Removal of Cypermethrin with Seaweed Gracilaria lemaneiformis

The removal of cypermethrin with a red macroalga, Gracilaria lemaneiformis, was studied under laboratory conditions. Results showed that the residue contents with G. lemaneiformis were significantly lower than those corresponding groups without the algal thalli after 96 h treatment. The removal rates decreased with increasing concentrations, which were about 50% without G. lemaneiformis after 96 h exposure, and increased to 89%, 73%, and 66% in flasks with G. lemaneiformis at the concentrations of 10, 100, and 1000 μg L-1, respectively. The amount of biosorption(absorption and adsorption) by G. lemaneiformis increased with the increasing concentration and exposure time. Adsorption was the main process for the removal by G. lemaneiformis, which accounted for 75%–97% of the total biosorption. However, biosorption only contributed 0.5%–19.3% to the total losses of cypermethrin, which was more efficient under the low concentration. Natural losses contributed the largest portion of losses, which was over 65% in all treatments during the experiment. The unknown pathway of removal, which might be the bio-decomposed by microorganisms attaching the algal thalli, also contributed a lot to the total removal. The results suggested that cultivation of G. lemaneiformis could significantly remove cypermethrin, especially at low concentrations, and large-scale cultivation of G. lemaneiformis has considerable potential of biorestoration of eutrophic and cypermethrin-polluted coastal sea areas.     

Ecotoxicity of Cadmium to Maize and Soybean Seedling in Black Soil

Ecotoxicity and bioavailability of cadmium(Cd)to the maize(Zea mays L.)and the soybean(Glycine max(L.)Merr.)were investigated by acute toxicity experiment in the laboratory with black soil.Ecotoxicity and bioavailability of Cd were quantified by calculating the median effective concentration(EC50)and bioaccumulation factor(BAF).The measurement endpoints used were seed germination and seedling growth(shoot and root).The results showed that concentrations of Cd in the soil had adverse effect on the growth of roots and shoots.Seed germination was not the sensitive indicator for the ecotoxicity of Cd in the soil,while the growth of roots was the most sensitive measurement endpoint.Absorbability and transport of Cd in plants depended on the test crop species and Cd concentrations in the soil.The maize retains more Cd in its roots,while the soybean transports more Cd to the shoots from roots.     

Distribution and elimination of Norfloxacin in Fenneropenaeus chinensis larvae

This study examined the distribution and elimination of Norfloxacin(NFLX) in Fenneropenaeus chinensis ovary and egg and newly hatched larvae.Mature parental shrimp were exposed to 4 or 10 mg L 1NFLX for 2 or 5 d.Ovary and eggs of the shrimp were sampled after spawning in order to detect NFLX residue using high-performance liquid chromatography(HPLC).Results showed that NFLX residue accumulated in F.chinensis eggs after the parental exposure,with the highest residue detected in ovary.To examine the fate of NFLX residue in larvae,we further determined the concentration of NFLX residue in F.chinensis eggs and larvae at 4 different developmental stages after 24-h exposure.From the newly metamorphosed larvae(0 h post-metamorphosis,h.p.m),samples were taken at different time intervals to 72 h.p.m.HPLC assay showed that the concentrations of NFLX residue in zoea exposed to 4 and 10 mg L 1NFLX were the highest at 1.5 h,i.e.,0.332 and 0.454 μg g 1,respectively.At the two NFLX exposure levels,the elimination time of half NFLX(half life) in nauplius was 45.36 and 49.85 h,respectively,followed by that in zoea(31.68 and 33.13 h),mysis larvae(42.24 and 47.28 h) and postlarvae(24.48 and 30.96 h).Both NFLX exposure levels had a germicidal effect.The distribution and elimination of NFLX residue in F.chinensis tissue,eggs and larvae correlated well with the drug exposure level.The disappearance of NFLX residue coincided with the larval growth,and the half-life of NFLX decreased with the larval development.     

The toxic effects of copper on larvae of the barnacle species Chthamalus challengeri

With the increased use of copper(Cu)-based antifouling(AF) paints,copper has become a potential threat to marine organisms. Experiments were performed to investigate the effects of copper on larvae of the barnacle Chthamalus challengeri. These experiments attempted to identify a more sensitive index to monitor copper pollution in marine environments. The 24 h LC5 0 ranged from 156.07 μg/L(nauplius II) to 817.15 μg/L(cypris) and the no observed effect concentration(NOEC) ranged from 81.75 μg/L(nauplius II) to 571.04 μg/L(cypris). The cypris settlement rate declined significantly when copper concentrations ≥135 μg/L. No cypris was found in the copper concentration of 60 and 75 μg/L treatment groups stressed for 22 d. Nauplius II moulting was not affected by exposure to copper for 24 h; however,when extended to 48 h,the percent moulted in 75 μg/L treatment group was declined to 37.12%,which was significant lower( P 0.05) than that in the control group. The phototaxis of nauplius II decreased significantly when copper concentrations ≥45 μg/L. Despite an initial significant increase at copper concentrations of 30 μg/L,ammonia excretion rate decreased when copper concentrations ≥60 μg/L. These results suggested that:(1) nauplius II could not develop to the cypris when the copper concentration ≥60 μg/L;(2) cypris settlement is more susceptible to copper than cypris mortality rate;(3) nauplius II is the most sensitive larval stage;(4) nauplius II ammonia excretion rate is the most sensitive index to copper and might be as the indicator for copper pollution monitoring.     

Ingestion of Domoic Acid and Its Impact on King Scallop（Pecten maximus,Linnaeus 1758）

A simple method for spiking formulated feed with domoic acid （DA） was developed in this study. DA feed was prepared by mixing 0.15 mL 100 μg mL-1 DA with 0.1 g formulated feed, and drying the mixture at room temperature for 2 h. The prepared DA feed contained 0.19 pg DA per particle. Of the added DA, 46.72% was retained in the feed. Relatively high DA retention （about 50%） was recorded after DA feed was soaked in water for 2h. Exposure to DA feed for 7d did not cause the increase of tissue DA level of adult king scallop （Pecten maximus） significantly in 60 d. The increase of their gonad index after DA exposure was not significantly different from the control. No significant change in DA level was found in spermary, ovary or fertilized eggs after DA exposure. These results indicated that DA excretion may be more efficient than DA accumulation under the current experimental conditions, and the mechanism of domoic acid incorporation in P. maximus may involve intracellular biotransformation.     

Generation of Reactive Oxygen Species in Different Fractions of the Coelomocytes of Holothurian Eupentacta fraudatrix in Response to the Thermostable Toxin of Yersinia pseudotuberculosis in Vitro

Pure fraction (92% - 95%) of phagocytes (FP) and a mixture of amoebocytes (62%) and morula cells (38 % )-FPMC- of the holothurian Eupentacta fraudatrix ( Holothuroidea,Dendrochirota ) were obtained by using ficoll-verographine step gradient. Basal production of reactive oxygen species (ROS) in FP quantified by using reduction of nitroblue tetrazolium (NBT) was more than twice that in FPMC. Thermostable toxin of Yersinia pseudotuberculosis (TST) at differentconcentrations ( 0.2; 0.5; 2.5 μg/ml, but not 0.1 μg/ml) stimulated NBT reduction in FPMC after 24 h incubation. In FP, TST at concentrations of 0.1 and 0.2 μg/ml inhibited and at concentrations of 0.5 and 2.5 μg/ml stimulated NBT reduction after 24 h incubation. Maximal effect was observed in FP and FPMC at TST concentrations of 0.5 and 0.2 μg/ml, respectively. Addition of catalase (0.7 μg/ml) to the cells treated with TST (2.5 μg/ml) was followed by a decrease in NBT reduction compared to that under toxin treatment alone. TST stimulated superoxide dismutase activity in ccncentration-dependent manner (maximum at 0.5 μg/ml concentration in FP) after 24 htreatment, and this stimulation was prevented by a commercial catalase. Plant lectin concanavalin A stimulated NBT reduction more than 5-fold in FPMC compared to the control. With addition of TST, lectin stimulated ROS to lesser extent than that with lectin alone. When catalase, TST, and lectin were added into the FPMC simultaneously, ROS increase was similar to that under lectin treatment alone. On the whole, data obtained indicated that ROS generation in holothurian coelomocytes especially occurs in both stimulated and not stimulated phagocytes, and that changes in ROS production by these cells may be one of the mechanisms of antibacterial protection of holothurians.     

Bioaccumulation of Pb<Superscript>2+</Superscript> and its effects on growth,morphology and pigment contents of <Emphasis Type="Italic">Spirulina</Emphasis> (<Emphasis Type="Italic">Arthrospira</Emphasis>) <Emphasis Type="Italic">platensis</Emphasis>

A laboratory experiment was conducted to assess the bioaccumulation of Pb²⁺ and its effects on growth, morphology and pigment contents of Spirulina (Arthrospira) platensis. The specimen cultured in Zarrouk liquid medium was treated with various initial metal concentrations (0, 5, 10, 30, 50 and 100 μg mL⁻¹). The growth of S. platensis was adversely affected by Pb²⁺ at high concentrations (30, 50 and 100 μg mL⁻¹). However, at low concentrations (5 μg mL⁻¹), Pb²⁺ could stimulate its growth slightly. The pigment contents (chlorophyll α and β carotene) were decreased in a dose-dependent manner. The highest reductions (67% and 53% respectively in chlorophyll α and β carotene) were observed in 100 μg mL⁻¹ treatment group. The LC₅₀ (96 h) of Pb²⁺ was measured as 75.34 μg mL⁻¹. Apart from a few cases of filament breakages at elevated concentrations (50 and 100 μg mL⁻¹), morphological abnormalities are not specific. Metal bioaccumulation increased with Pb²⁺ concentrations, but decreased with exposure time. The maximum accumulated amount was 188 mg g⁻¹ dry weight. The bioconcentration factor (BCF) reached to a peak at day 2, followed by a gradual reduction for all the exposure concentrations. S. platensis is able to tolerate considerably high Pb²⁺ concentrations. Consequently it can be used as a potential species to remove heavy metal from contaminated waters.     

Effects of heavy metals (Pb2+ and Cd2+) on the ultrastructure, growth and pigment contents of the unicellular cyanobacterium Synechocystis sp. PCC 6803

The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb2+ and Cd2+ (0, 0.5, 1, 2, 4, 6 and 8 mg/L). The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 ceils became evident with the increased (>4 mg/L Pb2+) metal concentration. The photosynthetic apparatus (thylakoid membranes) were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration (8 mg/L pb2+), the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations (0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L Pb2+), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll a, βcarotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb2+, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll a, β carotene and phycocyanin respectively over the control. Corresponding reductions for the same Cd2+concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC50) values (3.47 mg/L Cd2+ and 12.11 mg/L Pb2+) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd2+ than Pb2+.     

Effects of heavy metals （Pb^2＋ and Cd^2＋） on the ultrastructure, growth and pigment contents of the unicellular cyanobacterium Synechocystis sp. PCC 6803

The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb²⁺ and Cd²⁺ (0, 0.5, 1, 2, 4, 6 and 8 mg/L). The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased (>4 mg/L Pb²⁺) metal concentration. The photosynthetic apparatus (thylakoid membranes) were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration (8 mg/L Pb²⁺), the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations (0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L Pb²⁺), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll α, β carotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb²⁺, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll α, β carotene and phycocyanin respectively over the control. Corresponding reductions for the same Cd²⁺concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC₅₀) values (3.47 mg/L Cd²⁺ and 12.11 mg/L Pb²⁺) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd²⁺ than Pb²⁺. Supported by the Chinese Scholarship Council     

4种重金属离子及其混合液对壶状臂尾轮虫的急性毒性试验

研究了铜、锌、镉、铅离子及其混合液对壶状臂尾轮虫(Brachionusurceus)的急性毒性影响,实验结果表明,Cu2+、Zn2+、Cd2+、Pb2+对壶状臂尾轮虫的24hLC50分别为83 .18μg/L、4 .27mg/L、7. 60mg/L、23 44mg/L,将4种离子按照质量比120∶20∶6∶1的比例配成混合金属溶液,其混合液对壶状臂尾轮虫的24hLC50为0 42mg/L。     

CpG-DNA对异育银鲫(Carassius auratus gibelio)免疫细胞活性的诱导作用

探索含CpG基序的未甲基化寡脱氧核苷酸(ODNs)对鱼类的免疫调节,将人工合成的鱼免疫刺激剂寡脱氧核苷酸序列1670(含一个CpG基序)、1670-D(含两个CpG基序)加入到离体培养的异育银鲫的头肾巨噬细胞和外周血白细胞中,采用MTT法测定其对鲤外周血白细胞增殖的影响,NBT还原法和Griess试剂显色法测定对头肾巨噬细胞的呼吸爆发的影响。结果显示,1670可显著(浓度为20.0μg·mL-1)或极显著(浓度为0.1、1.0、10.0μg·mL-1)提高异育银鲫巨噬细胞的氧呼吸爆发活性,但对氮呼吸爆发活性和外周血白细胞的增殖无显著作用;1670-D可显著(浓度为0.1μg·mL-1)或极显著(浓度为1.0、10.0、20.0μg·mL-1)增强异育银鲫巨噬细胞的氧呼吸爆发活性和外周血白细胞的增殖,且可极显著(浓度为1.0、10.0、20.0μg·mL-1)提高异育银鲫巨噬细胞的氮呼吸爆发活性。表明特定序列、特定作用剂量的CpG-DNA可增强体外培养的异育银鲫巨噬细胞和外周血白细胞的非特异性免疫应答。     

TraceeElements in Leccinum scabrum mushrooms and topsoils from Kłodzka Dale in Sudety Mountains, Poland

In the current study, we determined concentrations and transfer rates of Ag, Al, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, Pb, P, Rb, Sr and Zn to Brown Birch Scaber Stalks (Leccinum scabrum) mushrooms emerged in the area of Sudety Mountains (Sudetes) in Poland. Fruiting bodies and topsoil samples beneath L. scabrum were collected form the K odzka Dale. The trace elements were determined using validated method and inductively coupled plasma - atomic emission spectroscopy (ICP-AES) for final measurement. Mushrooms contained Ag, Cr, Hg, Co, Ni and Sr at < 1.0 μg/g dry weight; Ba and Pb at ~1.0 μg/g dw; Cd at < 5 μg/g dw; Cu and Mn at > 10 μg/g dw; Al and Ca at ~100 μg/g dw; Fe, Na, Rb and Zn at 100 to 500 μg/g dw, Mg at ~1,000 μg/g dw; P at ~5,000 μg/g dw and K at ~30,000 μg/g dw. Ca, Mn and Ni were nearly equally distributed between stipes and caps; stipes compared to caps were enriched in Ba, Na and Sr, while caps were enriched in Ag, Al, Cd, Co, Cr, Cu, Fe, K, P, Pb, Rb and Zn. The values of bioconcentration factor (BCF) varied highly depending on chemical element and were >1 for Ag, Cd, Cu, K, Mg, Na, P, Rb and Zn, while <1 for Al, Ba, Ca, Co, Cr, Fe, Mn, Ni, Pb and Sr. Topsoil showed elevated content of lead and mean concentration was 99 ± 32 μg/g dw, while cadmium was at 0.41 ± 0.15 and those two highly toxic to human elements occurred in edible caps of L. scabrum at 4.5 ± 2.2 and 2.9 ± 2.0 μg/g dw, respectively.     

培育密度、换水量和倒池培育对珠母贝浮游幼虫生长发育和存活的影响

以室内育苗池或玻璃缸养殖方式,研究了不同培育密度、换水量和倒池培育对珠母贝浮游幼虫生长发育和存活的影响。结果表明:当培育密度为0.8、1.2只/mL,直线铰合期时间分别为7.00、7.40d,日均生长为5.16、4.87μm,壳顶期时间分别为10.76d和10.77d,眼点期密度为0.58只·mL-1和0.60只·mL-1,差异不显著;日换水量分别为40%、60%、80%,直线铰合期时间、壳顶期时间、日均生长及眼点期密度差异不显著,日换水量40%能满足幼虫的生长和存活;壳顶初期进行倒池培育,能促进浮游幼虫的生长发育和提高其存活率。     

Acid volatile sulfide and simultaneously extracted metals in superficial sediments from Baihua Lake, China

The bioavailability of five divalent cationic heavy metals(Pb,Cd,Cu,Zn and Ni) in 10 superficial sediment samples from Baihua Lake was assessed based on the molar ratio of simultaneously extracted metals(SEMs) to acid volatile sulfide(AVS).Atomic absorption spectrometry(AAS) and X-ray powder diffraction(XRD) were used to determine the heavy metal concentrations and examine the mineralogy of the crystalline phases,respectively.The AVS loadings in sediments from Baihua Lake ranged from 64.30 to 350.08 μmol/g(dry weight).The corresponding SEM levels for the sampling sites varied from 1.770 to 14.660 μmol/g.The molar ratio of SEMs to AVS ranged from 0.014 to 0.084 with a mean value of 0.034.The XRD analysis also confirmed the presence of some metal sulfides in sediments from Baihua Lake.The SEMs/AVS ratios for all sampling sites were significantly lower than 1.0,indicating that AVS in the sediments was sufficient to bind the five heavy metals;thus,these heavy metals are currently not significantly bioavailable to benthic organisms.Comparing the SEMs results to published guideline values for metal toxicity to benthic organisms in sediments,however,suggests that Zn and Ni pose a risk at some sampling locations in Baihua Lake.