Predictability of Copper, Irgarol, and Diuron Combined Effects on Sea Urchin Paracentrotus lividus

The aim of this work was to investigate the mixture toxicity of Irgarol (2-methylthio-4-t-butylamino-6-cyclopropylamino-s-triazine), Diuron (3-(3,4-dichlorophenyl)-1,1-dimethylurea), and copper upon the sea urchin Paracentrotus lividus and to compare the observed data with the predictions derived from approaches of Concentration Addition (CA) and Independent Action (IA). Copper spermiotoxicity was more sensitive (EC50 = 0.018 mg/L) than embryotoxicity (EC50 = 0.046 mg/L). The offspring malformations were mainly P1 type (skeletal alterations) in both cases, probably because copper competes to fix Ca²⁺. Irgarol and Diuron toxicity has been previously investigated. EC50 mixture embryotoxicity showed an EC50 of 1.79 mg/L, whereas spermiotoxicity mixture effects were lower than 11%. Both CA and IA modeling approaches failed to predict accurately mixture toxicity. For embryotoxicity, the IA model overestimated the mixture toxicity at effect levels of <80%. CA does not represent the worst-case approach showing values lower than IA (embryotoxicity) or similar (spermiotoxicity).     

Simultaneous determination of thiamin and riboflavin in the sea urchin, Paracentrotus lividus, by high-performance liquid chromatography

Thiamin and riboflavin were determined in the sea urchin, Paracentrotus lividus, by reversed-phase liquid chromatography with fluorescence detection and a mobile phase consisting of 72% 0.005 M NH(4)OAc and 28% MeOH at a flow rate of 1.450 ml/min. The total analysis time never exceeded 20 min. After acid hydrolysis and enzymatic digestion, thiamin was oxidized to thiochrome and the oxidized fraction was purified and concentrated by solid-phase extraction. Limits of detection were 9 pg/ml and 80 pg/ml for thiamin and riboflavin, respectively. Good results were obtained with respect to repeatability (RSD%<2.4) and recovery (>91.3%).     

Toxicity of tributyltin and triphenyltin to early life-stages of Paracentrotus lividus (Echinodermata: Echinoidea)

Sperm cell and embryo toxicity tests using the Mediterranean sea urchin Paracentrotus lividus were performed to assess the toxicity of tributyltin chloride, bis(tributyltin)oxide, triphenyltin acetate, and triphenyltin hydroxide. Toxicity values (mean effective concentration [EC50]) ranged from 2.97 to 18.5 microg/L for sperm cells and from 1.11 to 2.62 microg/L for embryos. For sperm cells, the toxicity of the two tributyl compounds was significantly greater than that of two triphenyl compounds; for embryos, the triphenyl compounds appeared to be more toxic. Study of embryotoxic effects highlighted closely concentration-dependent damages, the most sensitive stages corresponding to the crucial phases of differentiation (gastrula and prisma). Both EC50 and no-observed-effect concentration values for the four organotin compounds are similar to those reported in the literature for early life stages of other marine organisms.     

Arsenic Exposure Affects Embryo Development of Sea Urchin, Paracentrotus lividus (Lamarck, 1816)

Toxicity tests were performed with embryos of Paracentrotus lividus to investigate the toxicological effect of two arsenic species: arsenate (As^V), expected to be more toxic, and dimethyl-arsinate (DMA) expected to be less toxic. Exposures to toxicants were performed at different developmental stages in order to identify the most sensitive phase of embryological development. Statistical analysis revealed a high significance of each factor (Molecule, Concentration and Time of exposure) and their interaction for the dependent variable “Percentage of normal-shaped plutei”. In particular, the 8 cell stage was the most sensitive to arsenic; at a concentration of 50 μg L^?1 DMA proved to be more toxic than As^V, resulting in nearly 50 % of normal-shaped plutei against the 74 % recorded for As^V. Starting the administration of arsenic at the morula stage, arsenate proved to be significantly more toxic when compared to DMA.     

Sulfide as a confounding factor in toxicity tests with the sea urchin Paracentrotus lividus: comparisons with chemical analysis data

Sperm cell and embryo toxicity tests with the sea urchin Paracentrotus lividus were performed to assess the toxicity of sulfide, which is considered a confounding factor in toxicity tests. For improved information on the sensitivity of these methods to sulfide, experiments were performed in the same aerobic conditions used for testing environmental samples, with sulfide concentrations being monitored at the same time by cathodic stripping voltammetry. New toxicity data for sulfide expressed as median effective concentration (EC50) and no-observed-effect concentration (NOEC) are reported. The EC50 value for the embryo toxicity test (total sulfide at 0.43 mg/L) was three times lower than for the sperm cell test (total sulfide at 1.20 mg/L), and the NOEC values were similar (on the order of total sulfide at 10(-1) mg/L) for both tests. The decrease in sulfide concentration during the bioassay as a consequence of possible oxidation of sulfide by dissolved oxygen was determined by voltammetric analysis, indicating a half-life of about 50 min in the presence of gametes. To check the influence of sulfide concentrations on toxicity effects in real samples, toxicity (with the sperm cell toxicity test) and chemical analyses also were performed in pore-water samples collected with an in situ sampler in sediments of the Lagoon of Venice (Italy). A highly positive correlation between increased acute toxicity and increased sulfide concentration was found. Examination of data revealed that sulfide is a real confounding factor in toxicity testing in anoxic environmental samples containing concentrations above the sensitivity limit of the method.     

Toxicity of heavy metals using sperm cell and embryo toxicity bioassays with Paracentrotus lividus (Echinodermata: Echinoidea): comparisons with exposure concentrations in the Lagoon of Venice,Italy

Sperm cell and embryo toxicity tests using the Mediterranean sea urchin Paracentrotus lividus Lmk were performed to assess the toxicity of As3+, Cd2+, Cr3+, Ni2+, Pb2+, Cu2+, Zn2+, and Hg2+. The aim of this study was to improve information about the comparative sensitivity of sea urchin bioassays to the heavy metals, which are an important cause of contamination in the ecosystem of the Lagoon of Venice. Considering the data in mM/L, the order of toxicity is Hg2+ > Cu2+ > Zn2+ > As3+ > or = Cr3+ > or = Cd2+ > or = Pb2+ > or = Ni2+ for the sperm cell test and Hg2+ > or = Pb2+ > Cu2+ > Zn2+ > Cd2+ > Ni2+ > As3+ > or = Cr3+ for the embryo test. New toxicity data for metals expressed as median effective concentration (EC50) and no-observed-effect concentration (NOEC) are reported for the Mediterranean species. Accurate observations of embryotoxic effects at increasing metal concentrations were done, detecting some different behaviors in metal toxicity. Toxicity data compared with water column and pore-water concentrations recorded in the Lagoon of Venice (Italy) demonstrate the potential ability of bioassays using sea urchin to detect important contaminants in this ecosystem.     

Comparison of Methods to Obtain a Liquid Phase in Marine Sediment Toxicity Bioassays with Paracentrotus lividus Sea Urchin Embryos

Sediment toxicity bioassays using planktonic organisms are widespread tools in environmental quality assessment, but they have limitations because the method for extracting and diluting the liquid phase may affect the final toxicity. The present study compares the toxicity on sea urchin embryos and larvae of elutriates and pore water obtained from intertidal sediments by various methods. The effects of mixing method, dilution protocol, aeration, and pH adjustment of the pore water are discussed. For a quantitative assessment of toxicity, a posteriori dilution of the elutriate yielded results comparable to mixing the equivalent proportions of sediment and sea water. The highest sensitivity was found by using pore water. Rotator mixing in air-tight closed flasks was a second-best choice. In both cases the geochemical parameters of the liquid phases obtained were not suitable for the incubation of water-column organisms. To avoid the environmentally irrelevant toxicity caused by these natural conditions typical of highly reduced sediments, gentle aeration or dilution with O₂-saturated control sea water is necessary prior to testing, despite the unavoidable changes in redox conditions. Measuring length of 48-h-old larvae improves the sensitivity and the discriminating power of the test. Received: 17 March 2001/Accepted: 20 July 2001     

Toxic Effects of Irgarol and Diuron on Sea Urchin Paracentrotus lividus Early Development, Fertilization, and Offspring Quality

Irgarol and Diuron are the most representative “organic booster biocides” that replaced organotin compounds in antifouling paints. It cannot be assumed beforehand that their use will have no environmental impact: more ecotoxicological data and a significant environmental monitoring are required. Spermio and embryotoxicities of the biocides Irgarol and Diuron were investigated on Paracentrotus lividus, the dominant echinoid species of the Mediterranean Sea. Spermiotoxicity was studied by assessing the effects of sperm exposure on fertilization rate as well as on the induction of transmissible damages to the offspring. Embryotoxicity was studied by assessing the developmental defects in the exposed larvae. The experimental results show a Diuron EC50 of 2.39 (± 0.21) mg/L with a NOEL of 0.25 mg/L for embryos, and of 5.09 (± 0.45) mg/L with a NOEL of 0.5 mg/L for sperms, respectively. Data obtained from the embryotoxicity test on Irgarol [EC50 0.99 (± 0.69) mg/L] are of the same order of magnitude as the literature data about Japanese urchins. Spermiotoxicity tests show an Irgarol EC50 of 9.04 (± 0.45) mg/L with a NOEL of 0.1 mg/L. These data show the different sensitivities of the two tests: embryos are more sensitive than sperms for both the tested chemicals and Diuron seems to be the less toxic one. Moreover, as a major output of the experimental work, tested herbicides exert transmissible damage to spermatozoa evidenced by larval malformations in the offspring, mainly P1 type (skeletal alterations). The comparison of the endpoints results offers an interesting indication of a probable different mode of action (Irgarol seems to interact with calcium homeostasis) of the two biocides.     

Shewanella harboring antimicrobial and copper resistance genes in sea urchins (Paracentrotus lividus) from the Crozon peninsula (Brittany,France)

Shewanella is a genus of aquatic non-fermenting Gram-negative bacteria with increasing numbers of reports of infections in humans and appearance of antimicrobial resistant strains. Cases of infection show a relatively strong association with seafood consumption or exposure to seawater. This study aimed to analyze Shewanella spp. isolated from the sea urchin Paracentrotus lividus collected from the Crozon peninsula (France) with the intention of obtaining insights into the role of this genus as a reservoir of antimicrobial and heavy metal resistance genes. Five among seven Shewanella isolates were resistant to antimicrobials, mainly to broad spectrum beta-lactams. Four isolates displayed multiple resistance to at least three of these antimicrobial classes: broad spectrum beta-lactams, aminoglycosides, macrolide, quinolones and/or tetracycline. Three antimicrobial resistance genes were detected in just one isolate encoding resistance to beta-lactam (bla_SHV and bla_TEM-1) and macrolide (ermB). In addition, the copper resistance gene cusB, was observed in this isolate which is also a plasmid carrier. Another copper resistance encoding gene, copA, was found among the isolates. These results indicate that the multidrug-resistant (MDR) Shewanella isolates and resistance genes could be potential risks to public health, due to the carrying of these MDR bacteria by sea urchins through human consumption.     

Assessing the Toxicity of Chemical Compounds Associated With Land-Based Marine Fish Farms: The Sea Urchin Embryo Bioassay With Paracentrotus lividus and Arbacia lixula

In aquaculture, disinfection of facilities, prevention of fish diseases, and stimulation of fish growth are priority goals and the most important sources of toxic substances to the environment, together with excretory products from fish. In the present study, embryos of two species of sea urchin (Paracentrotus lividus and Arbacia lixula) were exposed to serial dilutions of six antibiotics (amoxicillin (AMOX), ampicillin, flumequine (FLU), oxytetracycline (OTC), streptomycin (ST), and sulfadiazine [SFD]) and two disinfectants (sodium hypochlorite (NaClO) and formaldehyde [CH(2)O]). Alterations in larval development were studied, and the effective concentrations (ECs) were calculated to evaluate the toxicity of the substances. Both species showed similar sensitivities to all substances tested. Disinfectants (EC(50) = 1.78 and 1.79 mg/l for CH(2)O; EC(50) = 10.15 and 11.1 mg/l for NaClO) were found to be more toxic than antibiotics. AMOX, OTC, and ST caused <20 % of alterations, even at the highest concentrations tested. FLU was the most toxic to P. lividus (EC(50) = 31.0 mg/l) and SFD to A. lixula (EC(50) = 12.7 mg/l). The sea urchin bioassay should be considered within toxicity assessment-monitoring plans because of the sensitivity of larvae to disinfectants.     

Developing Toxicity Scores for Embryotoxicity Tests on Elutriates with the Sea Urchin Paracentrotus lividus, the Oyster Crassostrea gigas, and the Mussel Mytilus galloprovincialis

Bioassays are routinely employed for sediment quality assessment. In order to be able to effectively use Bioassays responses in regulatory and management frameworks, toxicity scores, which rank toxicity data in defined classes that are continuous and difficult to interpret, should be reliable and suitable tools to support decisions about the presence or absence of toxicity in tested samples and on how toxic a sample is. A statistical approach is needed to define thresholds for toxicity scores. The Minimum Significance Difference (MSD) criterion allowed the evaluation of toxicity thresholds for each test-matrix and organism pair, based on large sets of experimental data. The MSD values were normalized with respect to the control, ranked in ascending order, and the 90th percentile was identified; the Toxicity Threshold (TT) was calculated by subtracting the 90th percentile from 100 and the Toxicity Limit (TL) was estimated as the percentage of control response multiplied by TT. Taking into account sample responses normalized with respect to control (S), when S > TL, the sample is considered nontoxic; when S 相似文献   

中药材中重金属的测定

采用微波消解样品、石墨炉原子吸收分光光度法测定植物药材中铅和镉 ,冷原子吸收分光光度法测定植物药材中的汞。用此方法测定了美国NBS15 70标准参考物质菠菜粉 ,以及 2 3种 2 6 4份植物药材。铅、镉、汞方法的平均回收率分别为 95 8%、97 5 %、99 2 % ;检出限 (LOD)分别为 0 0 0 6mg kg、0 2 0 8μg kg、0 0 2 6 μg kg;定量限 (LOQ)分别为 0 0 19mg kg、0 6 94μg kg、0 0 87μg kg。此方法同样适用于食物样品中铅、镉和汞的检测工作     

Dietary fibers and heavy metal retention in the rat

The metal-binding capacities of some gel-forming polysaccharides and other substances have been investigated in vitro in an attempt to relate their metal-binding properties to the retention of dietary Pb and Cd in vivo. In equilibrium dialysis systems, aqueous solutions of alginic acid, pectin, agar, and carrageenan (1 g fiber/100 ml) all bound Pb and Cd to varying degrees. Alginic acid had the greatest binding capacity for Pb (50 micrograms Pb bound/mg fiber) and carrageenan for Cd (9.3 micrograms Cd bound/mg fiber). Addition of any one of these fibers, or indulin or glucuronic acid to the diet increased the tissue retention of one or both of the metals. Only cellulose supplementation reduced the retention of both Pb and Cd. Carrageenan decreased that of Pb and increased that of Cd. In another experiment alginic acid was shown to increase Pb retention in rats even when present at fairly low dietary concentrations (1 g/kg).     

Naturally-occurring heavy metal binding proteins in invertebrates

Low molecular weight cadmium, copper, and zinc binding protein has been isolated from the musselMytilus edulis in the cadmium polluted areas of Corio Bay adjacent to Melbourne. Its molecular weight and other properties suggest that it belongs to the metallothionein family of metal binding proteins. Its synthesis may have been induced in the cyctoplasmic solutions of the cells as a regulatory mechanism to avoid intoxication by excess uptake of any of the metals.     

Assimilation and bioconcentration of Ag and Cd by the marine black bream after waterborne and dietary metal exposure

We determined the aqueous uptake and dietary assimilation of Cd and Ag by the marine black bream Acanthopagrus schlegeli following one to four weeks' exposure (or conditioning) to waterborne or dietary Cd or Ag at different concentrations. The concentrations of metals and metallothioneins (MT) in different tissues also were determined. The viscera contained the highest Ag, Cd, and MT concentrations after metal exposure. After exposure to waterborne metals, the metal and MT concentrations in the gills were higher than those in the remaining tissues (mainly muscles and bones), but this pattern was reversed following exposure to dietary metals. The assimilation efficiencies (AEs) of Cd and Ag ranged from 6 to 24% and 15 to 30%, respectively. The rate constant of uptake from the dissolved phase (ku) of Cd and Ag ranged from 2.2 to 7.5 and 8.0 to 31.7 L kg(-1) d(-l), respectively. In all the exposure experiments, the ku and AE increased with induced MT concentration and tissue metal concentration. Increasing metal accumulation may have been due to the increased available binding sites following the induction of MT in the fish. Furthermore, the MT induced by either Cd or Ag was not specific, but was able to bind with both metals and enhance bioaccumulation. Exposure to dissolved and dietary metals may increase metal accumulation, which potentially may lead to metal toxicity, although the fish may develop a tolerance to metals due to the apparent induction of MT.