Acute toxicity of organochlorine insecticide endosulfan to the giant freshwater prawn Macrobrochium rosenbergii

Endosulfan, an organochlorine pesticide, is highly toxic and effective at controlling pests in agriculture, horticulture, and public health programs. In this study, static bioassays were used to evaluate the toxicity of endosulfan to freshwater prawns( Macrobrachium rosenbergii) of various lengths(1.5±0.03,4±0.08, and 7±0.06 cm). Additionally, the activities of peroxidase(POD), acid phosphatase(ACP),alkaline phosphatase, acetylcholinesterase(AChE), and Na + /K +-ATPase were analyzed to refl ect the effects of endosulfan exposure. The 96 h LC 50 of endosulfan for prawns 1.5, 4, and 7 cm long were 1.86, 4.53,and 6.09μg/L, respectively, improved tolerance to endosulfan with growth. The POD activities of test organisms exposed to low concentrations of endosulfan were inhibited, indicating the presence of oxygen damaged tissue. Moreover, a notable decrease in AChE activity was observed due to overstimulation of neurotransmission, which might result in abnormal behavior. The effect caused by endosulfan on phosphatase production in the hepatopancreas of prawns 1.5, 4, and 7 cm long was different because the ability of nonspecifi c immune regulation increased with growth. The 96 h LC 50 values obtained in this study could be used in the formulation of water-quality criteria in China. Moreover, the changes in enzymes activities of M. rosenbergii under stress of endosulfan could be applied in the establishment of early warning indicators for bio-safety.     

Effect of cadmium on the defense response of Pacific oyster Crassostrea gigas to Listonella anguillarum challenge

Heavy metal pollution can affect the immune capability of organisms.We evaluated the effect of cadmium(Cd) on the defense responses of the Pacific oyster Crassostrea gigas to Listonella anguillarum challenge.The activities of several important defensive enzymes,including superoxide dismutase(SOD),glutathione peroxidase(GPx),acid phosphatase(ACP),Na+,K+-ATPase in gills and hepatopancreas,and phenoloxidase-like(POL) enzyme in hemolymph were assayed.In addition,the expression levels of several genes,including heat shock protein 90(HSP90),metallothionein(MT),and bactericidal/permeability increasing(BPI) protein were quantified by fluorescent quantitative PCR.The enzyme activities of SOD,ACP,POL,and GPx in hepatopancreas,and the expression of HSP90 were down-regulated,whereas GPx activity in the gill,Na+,K+-ATPase activities in both tissues,and MT expression was increased in Cdexposed oysters post L.anguillarum challenge.However,BPI expression was not significantly altered by co-stress of L.anguillarum infection and cadmium exposure.Our results suggest that cadmium exposure alters the oysters’ immune responses and energy metabolism following vibrio infection.     

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.     

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.     

Vitreoscilla hemoglobin gene (vgb) improves lutein production in Chlorella vulgaris

Vitreoscilla hemoglobin is an oxygen-binding protein that promotes oxygen delivery and reduces oxygen consumption under low oxygen conditions to increase the efficiency of cell respiration and metabolism. In this study, we introduced a Vitreoscilla hemoglobin gene （vgb） into Chlorella vulgaris by Agrobacterium tumefaciens-mediated transformation （ATMT）. PCR analysis confirmed that the vgb gene was successfully integrated into the Chlorella vulgaris genome. Analysis of biomass obtained in shake flasks revealed transformant biomass concentrations as high as 3.28 g/L, which was 38.81% higher than that of the wild-type strain. Lutein content of transformants also increased slightly. Further experiments recovered a maximum lutein yield of 2.91 mg/L from the transformants, which was 36.77% higher than that of the wild-type strain. The above results suggest that integrated expression of the vgb gene may improve cell growth and lutein yield in Chlorella vulgaris, with applications to lutein production from Chlorella during fermentation.     

Optimization of fermentation conditions for 1,3-propanediol production by marine Klebsiella pneumonia HSL4 using response surface methodology

The industrially important organic compound 1,3-propanediol(1,3-PDO) is mainly used as a building block for the production of various polymers. In the present study, response surface methodology protocol was followed to determine and optimize fermentation conditions for the maximum production of 1,3-PDO using marine-derived Klebsiella pneumoniae HSL4. Four nutritional supplements together with three independent culture conditions were optimized as follows: 29.3 g/L glycerol, 8.0 g/L K 2 HPO 4, 7.6 g/L(NH 4) 2 SO 4, 3.0 g/L KH 2 PO 4, pH 7.1, cultivation at 35°C for 12 h. Under the optimal conditions, a maximum 1,3-PDO concentration of 14.5 g/L, a productivity of 1.21 g/(L·h) and a conversion of glycerol of 0.49 g/g were obtained. In comparison with the control conditions, fermentation under the optimized conditions achieved an increase of 38.8% in 1,3-PDO concentration, 39.0% in productivity and 25.7% in glycerol conversion in flask. This enhancement trend was further confirmed when the fermentation was conducted in a 5-L fermentor. The optimized fermentation conditions could be an important basis for developing lowcost, large-scale methods for industrial production of 1,3-PDO in the future.     

The induction and extraction of metallothioneins inArtemia

A small amount of heavy metal binding protein, identified by BioRad Protein Assay, has been isolated from the adult brine shrimp,Artemia franciscanus. This protein has an apparent molecular weight between 6000 to 9000 dalton. a UV absorption peak at 260 instead of 280 nm like most proteins; and has high affinity towards binding with radioactive labeled¹⁰⁹Cd. These characteristics are similar to that of metallothioneins reported for many vertebrate and invertebrate, marine and terrestrial animals. After the brine shrimp is exposed to a small amount of Cd²⁺ for 24 h, a large amount of metallothionein can be isolated, showing the inducibility of this detoxifying protein in the adultArtemia in a short period of time.     

Comparison in copper accumulation and physiological responses of Gracilaria lemaneiformis and G. lichenoides (Rhodophyceae)

Heavy metal pollution has become a worldwide problem in aquaculture. We studied copper (Cu2+ ) accumulation and physiological responses of two red algae Gracilaria lemaneiformis and Gracilaria lichenoides from China under Cu2+ exposure of 0-500 μg/L in concentration. Compared with G. lemaneiformis, G. lichenoides was more capable in accumulating Cu2+ , specifically, more Cu2+ on extracellular side (cell wall) than on intracellular side (cytoplasm) and in cell organelles (especially chloroplast, cell nucleus, mitochondria, and ribosome). In addition, G. lichenoides contained more insoluble polysaccharide in cell wall, which might promote the extracellular Cu2+ -binding as an efficient barrier against metal toxicity. Conversely, G. lemaneiformis was more vulnerable than G. lichenoides to Cu 2+ toxin for decreases in growth, pigment (chlorophyll a, chlorophyll b, phycobiliprotein, and β-carotene) content, and photosynthetic activity. Moreover, more serious oxidative damages in G. lemaneiformis than in G. lichenoides, in accumulation of reactive oxidative species and malondialdehyde, and in electrolyte leakage, because of lower antioxidant enzyme (superoxide dismutase and glutathione reductase) activities. Therefore, G. lichenoides was less susceptible to Cu2+ stress than G. lemaneiformis.     

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     

Characterization of ribulose-1, 5-bisphosphate carboxylase/oxygenase and transcriptional analysis of its related genes in Saccharina japonica (Laminariales,Phaeophyta)

Saccharina japonica is a common macroalga in sublittoral communities of cold seawater environments,and consequently may have highly effi cient ribulose-1,5-bisphosphate carboxylase/oxygenase(Rubisco)activity for carbon assimilation.In our study,we cloned the full-length Rubisco gene from S.japonica(SJ-rbc).It contained an open reading frame for a large subunit gene(SJ-rbcL)of 1 467 bp,a small subunit gene(SJ-rbcS)of 420 bp,and a SJ-rbcL /S intergenic spacer of 269 bp.The deduced peptides of SJ-rbcL and SJ-rbcS were 488 and 139 amino acids with theoretical molecular weights and isoelectric points of 53.97 kDa,5.81 and 15.84 kDa,4.71,respectively.After induction with 1 mmol/L isopropyl-β-Dthiogalactopyranoside for 5 h and purifi cation by Ni 2+ affi nity chromatography,electrophoresis and western blot detection demonstrated successful expression of the 55 kDa SJ-rbcL protein.Real-time quantitative PCR showed that the mRNA levels of SJ-rbcL in gametophytes increased when transferred into normal growth conditions and exhibited diurnal variations: increased expression during the day but suppressed expression at night.This observation implied that Rubisco played a role in normal gametophytic growth and development.In juvenile sporophytes,mRNA levels of SJ-rbcL,carbonic anhydrase,Calvin-BensonBassham cycle-related enzyme,and chloroplast light-harvesting protein were remarkably increased under continuous light irradiance.Similarly,expression of these genes was up-regulated under blue light irradiance at 350 μmol/(m 2·s).Our results indicate that long-term white light and short-term blue light irradiance enhances juvenile sporophytic growth by synergistic effects of various photosynthetic elements.     

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.     

Impact of <Emphasis Type="Italic">Vibrio parahaemolyticus</Emphasis> and white spot syndrome virus (WSSV) co-infection on survival of penaeid shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

White spot syndrome virus (WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect Litopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus (WSSV+Vp), or we first infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection (Vp+WSSV). The eff ect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of Vibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction (PCR) method. LvMyD88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the diff erent mortality rates. Our results show that (1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection; (2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed significant increases in the numbers of Vibrio (P<0.05); (3) the WSSV copy numbers in the gills of the WSSV+Vp, Vp+WSSV, and the WSSV-alone groups increased from 10⁵ to 10⁷ /mg tissue 72, 96, and 144 h after infection, respectively. These results suggest that V. parahaemolyticus infection accelerated proliferation of WSSV in L. vannamei and vice versa. The combined accelerated proliferation of both V. parahaemolyticus and WSSV led to massive death of L. vannamei.     

Effects of dietary genistein on GH/IGF-I axis of Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

There is considerable concern that isoflavones, such as genistein in fish feed composed of soybean protein, aff ects somatic growth in fish. Our previous works demonstrated that 30 and 300 μg/g dietary genistein had no significant eff ect on growth performance in Nile tilapia (Oreochromis niloticus), but the higher level of genistein (3 000 μg/g) significantly depressed growth. This study was conducted to further examine the eff ects of dietary genistein on the endocrine disruption on growth hormone/insulin-like growth factor-I (GH/IGF-I) axis in Nile tilapia (O. niloticus). Juvenile fish were fed by hand twice daily to satiation with one of four isonitrogenous and isoenergetic diets, each containing either 0, 30, 300 or 3 000 μg/g genistein. Following an 8-week feeding period, plasma GH and IGF-I levels were investigated by radioimmunoassay and gene expression levels of gh, ghrelin, gnrhs, ghr, npy, npyrs, pacap, ghrs, igf-I, igf-Ir, and igfbp3 were examined by real-time PCR. The results show that no significant change in plasma GH and IGF-I levels in fish fed with diets containing 30 μg/g and 300 μg/g genistein. mRNA expression of genes along the GH/IGF-I axis remained unaff ected, except for igf-Ir, which was stimulated by the 300 μg/g genistein diet. While in fish fed the 3 000 μg/g genistein diet, the plasma GH and IGF-I levels decreased, and mRNA expression of gh, ghr2, npyr1, igf-I, and igf-Ir were also significantly depressed. In contrast, npy and igfbp3 mRNA expression were enhanced. This study provides convincing evidence for growth impediment by genistein by disturbing the GH/IGF-I axis in Nile tilapia O. niloticus.     

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 heavy metal ions (Cu2+, Pb2+ and Cd2+) on DNA damage of the gills, hemocytes and hepatopancreas of marine crab, Charybdis japonica

There are rising concerns about the hazardous effects of heavy metals on the environment. In this study, comet assay and DNA alkaline unwinding assay were conducted on the tissues (gills, hepatopancreas, and hemocytes) of Charybdis japonica in order to illustrate genotoxicity of three heavy metal ions (Cu2+, Pb2+, and Cd2+) on the marine crabs C. japonica. The crabs were exposed to Cu2+ (10, 50, and 100 ?g.L?1), Pb2+ (50, 250, and 500 ?g L?1) and Cd2+ (5, 25, and 50 ?g L?1), and the tissues were sampled at days 0.5, 1, 3, 6, 9, and 15. DNA alkaline unwinding assay was used for testing the DNA single strand break in gills and hepatopancreas and comet assay was employed for testing the DNA damage in hemocytes. The results showed that the DNA damage (F-value) of gills in the crabs exposed to the three heavy metals was decreased gradually during the exposure periods and there was a dose-time response relationship in certain time, suggesting that the levels of DNA single strand break in all the experimental groups increased significantly compared to the controls. Changes of F-value in hepatopancreas of the crabs exposed to the three heavy metals were similar to those in gills except that the peak values were found in the 500 ?g L?1 Pb2+ treatment group at day 3 and the 50 ?g L?1 Cd2+ treatment group at day 9. The ranks of DNA damage in gills and hepatopancreas induced by the three heavy metal ions (50 ?g L?1, day 15) were Cd2+ >Pb2+ >Cu2+ and Pb2+ >Cu2+ >Cd2+. The levels of DNA damage in gills were higher than those in hepatopancreas in the same experimental group. It can be concluded that indices of DNA damage can be used as the potential biomarkers of heavy metal pollution in marine environment.     

Residues of organochlorine pesticides in intertidal flat surface sediments from coastal zone of Jiangsu Province,China

Sixteen surface sediment samples were collected and analysed to evaluate the residues of organochlorine pesticides(OCPs) from intertidal flat in Jiangsu Province. Overall, 22 OCPs were detected with total concentrations of OCPs ranging widely from 0.96 to 12.14 ng/g(dry wt). Total hexachlorocyclohexane(HCH) and total dichlorodiphenyltrichloroethane(DDT) levels varied from 0.01 to 0.67 ng/g and from 0.23 to 4.85 ng/g, respectively. DDTs were the predominant compounds. The dominance of β-HCH indicated a history of HCH pollution. According to the ratios of( p, p'-DDD+ p, p'-DDE)/ p, p '-DDT and o, p ′-DDT/ p, p ′-DDT, new input of DDTs did not occur in most sites, and the main sources were historical usage of technical DDTs. OCPs such as dieldrin, endrin, p, p ′-DDD, and p, p ′-DDT exceeded the effects range low, showing adverse biological effects that would occasionally occur at some sites of the study area.     

The influence of cadmium on the antioxidant enzyme activities in polychaete Perinereis aibuhitensis Grube (Annelida: Polychaeta)

The infaunal polychaete Perinereis aibuhitensis Grube, distributed widely along Asian coasts and estuaries, is considered a useful animal model in ecotoxicological tests and a promising candidate in biomonitoring programs. This paper deals with the activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidases (GSH-Px) in infaunal polychaete P. aibuhitensis exposed to a series of sublethal water-bound cadmium (Cd) concentrations (0, 0.34, 1.72, 3.44, 6.89, and 17.22 mg L⁻¹) under a short-term exposure (1–8 d). The results indicate that the SOD and GSH-Px activities in P. aibuhitensis are stimulated first and then renewed to the original level. The CAT activity of worms decreases at an earlier exposure time but increases to the control values at a later exposure time. Our study suggests that Cd can interfere with the antioxidant defense system of P. aibuhitensis. However, the changes in antioxidant enzyme activities for this species do not show the best promise as biomarkers in Cd biomonitoring of estuarine and coastal zones because weak or non-dose-effect relationships between the antioxidant enzymes activities and Cd levels are found.