Application of the sea urchin sperm bioassay to sewage treatment efficiency and toxicity in marine waters

A newly standardized 60-minute sea urchin (and sand dollar) sperm bioassay was used to measure the toxicity of sewage at each stage in the treatment process. The sensitivity of the sperm bioassay was compared to the results of 48- to 96-hour bioassays with sea urchin and oyster embryos and crab zoeae in side-by-side tests of each sewage type. These data were then used to estimate the dilutions necessary to protect sensitive life stages of marine animals around a proposed sewage outfall in Puget Sound and to select the best bioassay procedure for a post-discharge program for monitoring water column toxicity.Results showed that very little acute toxicity was removed by the primary treatment process but that toxicity was greatly reduced by secondary treatment. A high degree of toxicity was reintroduced by chlorination but this source of toxicity was essentially eliminated by dechlorination. The simple and rapid sperm assay proved more sensitive to all stages of sewage than did the longer embryo and larval assays. Based on the results of the comparative testing and the ease, rapidity and year-round availability of the sea urchin sperm assay (sea urchins in winter, sand dollars in summer), this test was recommended for post-discharge monitoring of sewage effluent-related toxicity in the receiving waters.     

Identification of specific malformations of sea urchin larvae for toxicity assessment: Application to marine pisciculture effluents

Standard toxicity screening tests are useful tools in the management of impacted coastal ecosystems. To our knowledge, this is the first time that the sea urchin embryo development test has been used to evaluate the potential impact of effluents from land-based aquaculture farms in coastal areas. The toxicity of effluents from 8 land-based turbot farms was determined by calculating the percentage of abnormal larvae, according to two criteria: (a) standard, considering as normal pyramid-shaped larvae with differentiated components, and (b) skeletal, a new criterion that considers detailed skeletal characteristics. The skeletal criterion appeared to be more sensitive and enabled calculation of effective concentrations EC₅, EC₁₀, EC₂₀ and EC₅₀, unlike the classical criterion. Inclusion of the skeleton criterion in the sea urchin embryo development test may be useful for categorizing the relatively low toxicity of discharges from land-based marine fish farms. Further studies are encouraged to establish any causative relationships between pollutants and specific larval deformities.     

Stress-induced apoptosis in sea urchin embryogenesis

We find that early sea urchin embryos have the capability to induce programmed cell death, or apoptosis, in response to chemical and physical stress. Strongylocentrotus purpuratus embryos (fertilized, 4 cell, 16 cell, 64 cell, and early blastula) were exposed to known cytotoxins, in order to determine when apoptosis occurs naturally and in response to stress. Using cell permeability as an indicator of early stage apoptosis, caspase activation as a mid-stage indicator, and DNA fragmentation as a late stage indicator, we find that during the cleavage stage of embryogenesis apoptosis is almost completely absent. However, a statistically significant (p<0.001) rise in apoptosis in stressed embryos is evident around 24 h after fertilization, during the early blastula stage and shortly after hatching. Before this stage, exposed embryos show no statistically significant increases in apoptosis in comparison to the controls. This pattern of apoptosis in development is similar to that seen in lower vertebrate models in which stress-induced apoptosis occurs only around the mid-blastula transition. We conclude that apoptosis may be used to rid embryos of aberrant or damaged cells in early development, but this response is stage-dependant. Repair, rather than apoptosis, may be utilized during earlier stages, or alternatively, embryos exposed to such stressors may continue development with damaged cells and perhaps damaged DNA. Our continued studies will focus on these alternative hypotheses.     

Influence of salinity on fertilization and larval development toxicity tests with two species of sea urchin

Sea urchin embryo-larval development (ELD) and fertilization tests have been widely used in ecotoxicity studies and are included in regulatory frameworks. Biological processes occur naturally within a range of salinity that depends on the species considered. In an attempt to determine the optimum range of salinity, ELD and fertilization bioassays were performed at different salinities (15-40.5‰) with two species of Atlantic sea urchin: Arbacia lixula and Paracentrotus lividus. In the ELD assay, the optimum range of salinity was wider for A.?lixula (29-35.5‰) than for P.?lividus (29-33‰). In the fertilization assay with P.?lividus as a bioindicator species, the highest percentage of fertilization (90%) was obtained at salinities of between 29 and 33‰. More research on A.?lixula is required, since the fertilization success was below 60%. The results of the present study demonstrate that salinity may be a confounding factor in interpreting ELD test results.     

Cadmium toxicity to three species of estuarine invertebrates

Three species of estuarine invertebrates, Palaemonetes pugio (grass shrimp), Pagurus longicarpus (hermit crab) and Argopecten irradians (bay scallop), were exposed to Cd in flowing seawater at concentrations of 0·06, 0·12, 0·25, 0·5 and 1·0 mg/litre. Incipient LC₅₀ values of 0·53 and 0·07 mg/litre were estimated for bay scallop and hermit crab, respectively. The toxicity curve for grass shrimp had not stabilised, but the incipient LC₅₀ value was estimated to fall within a range of 0·2 to 0·3 mg/litre. Short-term response, as measured by time to 50% mortality at the highest Cd concentration, was 10, 21 and 23 days for the bay scallop, hermit crab and grass shrimp, respectively. Scallop growth was inhibited at all exposure concentrations with a measured 42-day EC₅₀ value of 0·078 mg/litre Cd. Byssal thread detachment precedes death in bay scallops. An EC₅₀ value of 0·54 mg/litre Cd for byssal detachment was measured on day 8 of the bioassay before appreciable mortality. This compared favourably with the incipient LC₅₀ value of 0·53 mg/litre Cd. Cadmium accumulation occurred at all concentrations in bay scallop and grass shrimp.     

The impact of rising sea temperature on innate immune parameters in the tropical subtidal sea urchin Lytechinus variegatus and the intertidal sea urchin Echinometra lucunter

Ocean temperatures are rising throughout the world, making it necessary to evaluate the impact of these temperature changes on sea urchins, which are well-known bioindicators. This study evaluated the effect of an increase in temperature on the immune response of the subtidal Lytechinus variegatus and the intertidal Echinometra lucunter sea urchins. Both species were exposed to 20 (control), 25 and 30 °C temperatures for 24 h, 2, 7 and 14 days. Counting of coelomocytes and assays on the phagocytic response, adhesion and spreading of coelomocytes were performed. Red and colorless sphere cells were considered biomarkers for heat stress. Moreover, a significant decrease in the phagocytic indices and a decrease in both cell adhesion and cell spreading were observed at 25 and 30 °C for L. variegatus. For E. lucunter, the only alteration observed was for the cell proportions. This report shows how different species of sea urchins respond immunologically to rising temperatures.     

Embryotoxicity of butyltin compounds to the sea urchin Paracentrotus lividus.

Tributyltin (TBT) has been widely employed in marine anti-fouling paints as a biocide, although it represents a serious risk, particularly in estuarine and coastal water/sediment ecosystems. In this study, the embryotoxic effects of TBT and its degradation products, dibutyltin (DBT) and monobutyltin (MBT), were analyzed during the development of the sea urchin Paracentrotus lividus from post-fertilization to the pluteus stage, to better clarify ecotoxicological impact. The embryotoxicity of butyltins is concentration-dependent and increases proportionally with number of butyl groups. Significant growth reduction was observed at TBT concentrations as low as 0.01 microgram l-1; 1 microgram l-1 was the maximum concentration allowing embryos to reach the pluteus stage at 48 h post-fertilization. Development was blocked at the morula or blastula stage with higher TBT concentrations. DBT and MBT are less toxic: slowed development and a decrease in pluteus size occurred at 10 micrograms l-1 DBT and 0.5 mg l-1 MBT. Effects on both skeletal deposition and blocked embryonic development are suggested to be due to the interference of organotin compounds with intracellular calcium homeostasis.     

Differences in trophic position among sympatric sea urchin species

Three species of sea urchin regularly co-occur in high abundances on subtidal rocky reefs in south-western Australia. We used two lines of evidence (stable isotope analysis and gut contents analysis), to test whether these species occupy different trophic positions. We looked at five discrete populations to test whether patterns were consistent. The gut contents of Heliocidaris erythrogramma contained almost exclusively fragments of macroalgae, and the δ¹⁵N of muscle was consistent with that expected for a herbivore. In contrast, the gut contents of Phyllacanthus irregularis and Centrostephanus tenuispinus contained a greater proportion of animal tissue, and the δ¹⁵N of muscle suggested that animal tissue was an important source of nutrition. Of the three co-occurring sea urchin species, one (H. erythrogramma) was ecologically dissimilar to the others and occupied a lower trophic position. This pattern was consistent among populations separated by up to 270 km in south-western Australia. Food resource partitioning might be one way in which these species are able to coexist.     

Participatory methodology used for sea urchin co-management in Barbados

This project aimed to develop co-management of exploited marine fishery resources in Barbados using the sea urchin fishery as a test case. The approach was to work with stakeholders, primarily the fishers, to establish a co-management mechanism that could be operated by the fishers themselves with technical and advisory support from the Fisheries Division. The project used Technology of Participation (ToP) methodology, developed by the Institute of Cultural Affairs (ICA). ToP methods used included Focused Conversation and Participatory Strategic Planning. To our knowledge, this methodology has not previously been applied in small-scale fisheries co-management. Fishers involvement was developed in stages: identification of groups of fishers in communities and a contact person for the group; dialogue with individuals and the small groups; discussion in larger groups to derive approaches to management; and full group participation to reach consensus regarding the most appropriate approach to management. Key persons identified in communities helped organize meetings to discuss the sea urchin fishery. From these community meetings, individuals were selected to take part in the strategic planning. Two vision meetings with separate groups of fishers, produced similar results. These groups were combined at a planning meeting, where fishers examined the blocks to achieving the vision, developed strategies to overcome the blocks, and an action plan to implement the strategies. Fishers and government officials concluded that the methodology had successfully facilitated the input of both parties and produced a workable, consensual approach.     

Sea urchin sperm bioassay for sewage and chlorinated seawater and its relation to fish bioassays

Bioassays were conducted with sea urchin and sand dollar sperm to determine the toxicity of chlorinated and unchlorinated sewage effluent and chlorinated and brominated seawater. The sperm cells were exposed to seawater dilutions of each toxicant for 5–15 min. The fertilisation of eggs served as the indicators of sperm viability. The effective concentrations which reduced fertilisation success by 50% (EC50) averaged 2·2 and 4·8% chlorinated and unchlorinated sewage in seawater, respectively. The sperm cells were extremely sensitive to chlorinated seawater at concentrations from 0·002 to 0·020 mg/litre total residual oxidant (TRO). Brominated seawater proved toxic to sperm in one test at 0·015 mg/litre TRO. Results of the sperm bioassays are compared with previous acute and chronic bioassays with fish.     

Relationships between sediment contamination and toxicity in San Francisco Bay

Sediment contamination and toxicity were monitored at 14 sites in San Francisco Bay between 1991 and 1996. Sediment contamination patterns were different in the major reaches of the Bay, and at each site. Several contaminants were consistently above concentrations previously associated with toxicity at most sites. Bulk sediment bioassays using the amphipod Eohaustorius estuarius and sediment elutriate bioassays using larval bivalves (Mytilus spp., Crassostrea gigas) also indicated different patterns of sediment toxicity in space and time. Sediments were most toxic to the amphipods at Redwood Creek (90% of the tests), and were toxic in at least half the tests conducted at five other sites. Sediment elutriates severely reduced normal bivalve larval development at the San Joaquin and Sacramento Rivers in all samples, but toxicity occurred in less than a third of the tests in the Central and South Bays. Toxicity could not be statistically related to seasonal freshwater flow or rainfall in the Bay, but seasonal variation in contaminant concentrations and toxicity was observed. Amphipod toxicity was inversely and significantly related to the mean effects range-median quotient, suggesting that cumulative concentrations of several contaminants were related to toxicity. Further analysis identified suites of specific contaminants at each site that were variably related to amphipod toxicity at each site. Chlordanes, cadmium, and silver were significantly related to amphipod survival in the North Bay. Seasonal patterns in low, and high molecular weight polycyclic aromatic hydrocarbons (PAHs) were related to toxicity at Alameda, and metals and PAHs were related to toxicity at Castro Cove. Larval bivalve toxicity was associated with metals in bulk sediments, but elutriate chemistry was not measured, and relationships with toxicity could not be examined. Hypotheses about effective concentrations of several individual contaminants and mixtures of contaminants were posed.     

Teratogenesis in Fundulus heteroclitus embryos exposed to a creosote-contaminated sediment extract and CYP1A inhibitors

The goal of these experiments was to explore the relationship between cytochrome P4501A (CYP1A) induction and the teratogenicity of sediments from the Atlantic Wood Industries Superfund site (Elizabeth River, VA) in Fundulus heteroclitus embryos. In these experiments we used embryos spawned from reference site adults to assess CYP1A activity and teratogenicity induced by aqueous Elizabeth River sediment extracts (ERSE). Embryo exposures to ERSE induced CYP1A activity and caused deformities, including pericardial edema, heart elongation and tail shortening. Co-exposures with various CYP1A inhibitors significantly decreased CYP1A activity and increased the teratogenicity of the sediment extract. Potential mechanisms for this increased toxicity are discussed herein.     

崎头洋附近群岛水域泥沙特征及悬沙来源分析

依据实测水文泥沙资料分析舟山群岛南部崎头洋附近海域的悬沙与底沙特征,建立该水域二维潮流数学模型,同时根据底沙及悬沙的粒径分布和潮流的运动路径对该海域的悬沙来源做进一步分析.指出崎头洋附近群岛海域的悬沙除部分来源于潮汐动力作用下的底沙起动再悬浮之外,主要是杭州湾附近灰鳖洋水域的泥沙在潮汐动力作用下经群岛西北方向水道进入群岛内部水域的,进而在潮动力作用下经由东南方向水道进入东海及象山港附近水域.     

海参、海胆养殖水体中多抗性细菌抗性基因的初步研究

抗药性细菌对生态环境、养殖业以及人类健康构成严重威胁。以大连海参、海胆养殖场多抗性细菌筛选为基础,采用分子生物学手段对40株抗性细菌进行了氯霉素、土霉素及氟氯霉素抗性基因检测,探讨抗药性分子遗传机理,从而为控制抗药性细菌的产生和传播提供研究基础。     

Comparative toxicities of chlorinated biphenyls on sea urchin egg fertilisation and embryogenesis

The aim of this study was to compare the biological actions of selected chlorinated biphenyls (CB's) (with different degrees of chlorine substitution), using the sea urchin test system, for their ability to affect fertilisation, embryogenesis and mitotic activity. Sea urchin embryos were exposed to the test agents throughout larval development or for shorter intervals (e.g. before or after hatching). Sperm or eggs were exposed before fertilisation, thereafter the contaminants were removed from embryo cultures. The results showed maximal activities from medium-chlorinated (tetra- and hexachloro-substituted) compounds, as compared to low- and high-chlorinated biphenyls (mono-, di- and octachloro-substituted), at nominal levels ranging from 10⁻⁶m to 10⁻⁵m. This held true for the different parameters and treatment schedules. Developmental defects were only detected following post hatching exposure of embryos to CB's, whereas pre-hatching exposure failed to reveal any embryological or cytogenetic abnormalities.     

Molecular characterization and expression of the SiUCP2 gene in sea urchin Strongylocentrotus intermedius

Uncoupling protein 2 (UCP2) is a proton transporter located in the inner mitochondrial membrane, and inhibits the formation of adenosine triphosphate and reacti...     

埕北海域底质沉积物粒度分布特征及其动力环境意义

通过分析181个埕北海域底质沉积物样品的粒度资料,并应用Flemming三角图示法对研究区进行沉积动力环境分区,结果表明:研究区底质沉积物主要为粉砂质砂、砂质粉砂、粉砂和黏土质粉砂4种类型,整体上由岸向海粒度呈现细-粗-细的分布特征,且呈条带状与岸线平行;粒度相对较粗,粒径频率曲线多以单峰为主,分选中等偏差,多为正偏;沉积动力环境分区表明该海域沉积物多以悬浮和跃移为主;粒度特征和沉积动力强度以6 m和12 m等深线为轴对称分布,12 m水深等值线两侧沉积物的分布类型和运移趋势主要受季节的风浪变化控制,6 m水深等值线两侧沉积物分布和运移趋势主要受波浪破碎的紊流作用控制。     

Characteristics of sediment transport in the Zhoushan Archipelago sea area

Based on the field data including tidal current,suspended sediment concentration,grain size of surface sediments,the transport mechanism and movement trend of sediments are analyzed using the method of flux decomposition and Grain Size Trend Analysis.The results show that the concentration and flux of suspended sediments decrease sharply from the Hangzhou Bay to the offshore area.The suspended sediment transport is mainly controlled by advection transport including average current transport and Stokes drift-induced transport and the gravitational circulation transport.The surface sediments are transported from the entrance of the Hangzhou Bay to the offshore area in general; meanwhile,the sediment transport has two obvious transport trends in the offshore area.The interaction of tidal currents,residual currents,the East China Sea coastal current,Taiwan Warm Current and wind waves appear to play important roles in the sediment transport.Furthermore,the sediment distribution and transport are significantly affected by the Zhoushan Archipelago.     

Bottom-up and top-down control of seagrass overgrazing by the sea urchin Tripneustes gratilla

The lack of top-down control on Tripneustes gratilla, a sea urchin commonly known to graze on seagrass, and the bottom-up control of its feeding preference, led to the overgrazing of seagrass meadows of the species Thalassodendron ciliatum in Changuu Island (Zanzibar Archipelago). The impact of overgrazing on seagrasses was assessed by mapping the presence of grazed versus non-grazed seagrass patches in the study site, while the top-down control on T. gratilla was assessed by measuring the abundance of its fish predators. The feeding preference and distribution of T. gratilla were characterized by calculating the electivity indexes for each seagrass species and measuring sea urchin density, respectively. Approximately half of the patches of T. ciliatum were overgrazed, while predatory fishes of T. gratilla were absent from the site. The Vanderploeg and Scavia's Relativized Electivity Index indicated that T. gratilla had a feeding preference for T. ciliatum, which was also supported by higher urchin densities within T. ciliatum dominated patches. Bottom-up control of grazing activity was observed by quantifying and analyzing morphological, nutritional, and the chemical defense traits of the seagrass in relation to feeding preference and urchin density. Feeding was positively correlated to the seagrass tissue C:P ratio (ρ = 0.9), whereas urchin density showed no correlations. The bottom-up control of the feeding preference and agglomeration of T. gratilla in T. ciliatum meadows, together with the lack of evidence of substantial top-down control and the long recovery time of T. ciliatum led to the overgrazing of this species at this site. Overgrazing, therefore, was shown to be the result of multiple factors ranging from the traits of the seagrass and feeding preference of T. gratilla, to the abundance of predators in this area.     

Teratogenesis in Fundulus heteroclitus embryos exposed to a creosote-contaminated sediment extract and CYP1A inhibitors

The goal of these experiments was to explore the relationship between cytochrome P4501A (CYP1A) induction and the teratogenicity of sediments from the Atlantic Wood Industries Superfund site (Elizabeth River, VA) in Fundulus heteroclitus embryos. In these experiments we used embryos spawned from reference site adults to assess CYP1A activity and teratogenicity induced by aqueous Elizabeth River sediment extracts (ERSE). Embryo exposures to ERSE induced CYP1A activity and caused deformities, including pericardial edema, heart elongation and tail shortening. Co-exposures with various CYP1A inhibitors significantly decreased CYP1A activity and increased the teratogenicity of the sediment extract. Potential mechanisms for this increased toxicity are discussed herein.