DNA damage and apoptosis in the mussel Mytilus galloprovincialis

The effects of known genotoxic substances (4-nitroquinoline-N-oxide, benzo[a]pyrene, teniposide, etoposide, cycloheximide, tributyltin) on human cells (FLC, HL-60) and on mussels were investigated. The correlations between formation of DNA strand breaks and DNA fragmentation characteristic for the process of apoptosis were estimated. Strand breaks induced by 4-nitroquinoline-N-oxide and benzo[a]pyrene did not correlate with DNA fragmentation detected in the process of apoptosis. Induction of internucleosomal DNA fragmentation in HL-60 cells was initiated by teniposide, etoposide and tributyltin, while in the gills of mussels this was detected only with tributyltin. Levels of DNA strand breaks in natural mussel populations, living at locations under the influence of urban and industrial wastes, do not mirror the apoptotic processes.     

Lysosomal reaction to xenobiotics in mussel hemocytes using BODIPY-FL-verapamil

Many cellular and sub-cellular biomarkers associated with mussel (Mytilus edulis) digestive gland and kidney have been characterised. The lysosomal compartment of these tissues have been recognised as being particularly sensitive, exhibiting pollutant induced responses which could be potentially used as a ‘biomarker’. However, relatively few studies have investigated the lysosomal response within molluscan hemocytes. This study was conducted to test whether lysosomal reactions, in live hemocytes isolated from mussels, can be used as a biomarker of pollutant exposure and deleterious effect. Lysosomal responses to a number of hydrocarbons, including anthracene and phenanthrene, and to the amphiphilic heterocylic chemical, chlorpromazine, were examined. The supravital dye neutral red (NR) was used to examine lysosomal membrane fragility, following xenobiotic exposure. NR was also used to verify the lysosomal compartment as the reported accumulation site of a new molecular probe, BODIPY-FL-verapamil (BFLV). The use of BFLV, with confocal laser microscopy and image analysis enabled visualisation and quantification of lysosomal distribution and perturbation. BFLV showed that exposure of molluscan hemocytes to xenobiotics (20 ppb–10 ppm) induced the formation of pathologically enlarged lysosomes. The internal trafficking of lysosomes was shown to be severely compromised after exposure to chlorpromazine. Exposed molluscan hemocytes exhibited significantly reduced lysosomal retention times, for neutral red. Preliminary data is presented demonstrating the opportunity for these non-destructive biomarker techniques to detect pollution gradients in situ.     

三种典型POPs对紫贻贝不同组织DNA损伤的比较研究

采用彗星电泳技术,比较研究了Aroclor 1254、BaP、DDT三种典型POPs对紫贻贝(Mytilus edulis)鳃、性腺、消化腺细胞的DNA损伤效应的差异,以期为进一步评价这三种典型POPs遗传毒理机制及其预警监测提供科学依据.结果表明,Tail DNA%是评价DNA损伤的理想指标,另外,在急性毒性试验中,...     

A carbon and nitrogen flux model of mussel digestive gland epithelial cells and their simulated response to pollutants

The mussel digestive gland epithelial cells provide a key interface between the organism and pollutants such as aromatic hydrocarbons. The simulation of their uptake and export mechanisms as well as an internal protein degradation pathway, and any subsequent disruption to any of them, has been undertaken. A computational model is described, which simulates the flow of carbon and nitrogen through a mussel's digestive cell. The model uses a compartmentalised view of the cell with inviolate 'pipelines' connecting each of the volume-variable partitions. Only the major physiological pathways relevant to the flow of either carbon or nitrogen or volume are modelled. Simulated response to hydrocarbon exposure is examined.     

Impact of the Sea Empress oil spill on lysosomal stability in mussel blood cells.

Coastal zones are among the most productive and vulnerable areas on the planet. An example of impact on these fragile environments was shown in the case of the "Sea Empress" oil tanker, which ran aground in the Bristol Channel in 1996, spilling 72,000 tons of "Forties" crude oil. The objective was to investigate the sub-lethal cellular pathology and tissue hydrocarbon contamination in marine mussel populations, 4 months after the initial spill, using the neutral red retention (NRR) assay for lysosomal stability in blood cells. NRR was reduced in mussels, and indicative of cell injury, from the two sites closest to the spill in comparison with more distant and reference sites. Lysosomal stability was inversely correlated with polycyclic aromatic hydrocarbon concentrations in mussel tissues. Reduced lysosomal stability has previously been shown to contribute to impaired immunocompetence and to autophagic loss of body tissues. The use of this type of technique is discussed in the context of cost-effective, ecotoxicological tools for Integrated Coastal Zone Management.     

Cellular responses in the mussel Mytilus edulis following exposure to diesel oil emulsions: Reproductive and nutrient storage cells

The objectives of this study were to examine, and quantify with the aid of stereological techniques,^1,2 the effects of exposure to hydrocarbons on the reproductive and nutrient storage cell systems in the marine mussel, Mytilus edulis. In addition, the capacity for recovery of these cell systems was quantified following a period of hydrocarbon depuration. The second phase of these investigations examined the significance of nutritional reserves at the time of exposure to hydrocarbons and how this may influence the animal's ability to survive the insult. The results (Table 1) indicated that, first, hydrocarbons had a deleterious effect on the nutritional storage cells leading to reduced fecundity and oocyte atresia (degeneration). Given a period of depuration, however, there were indications of a recovery in both the reproductive and storage cell systems. Second, the ability of mussels to survive hydrocarbon exposure was dependent upon the nutrient reserves at the onset of exposure.     

Antioxidant and pro-oxidant challenge of tannic acid in mussel hemocytes exposed to cadmium

The present study investigates the antioxidant and pro-oxidant behavior of tannic acid (TA) in hemocytes of mussel Mytilus galloprovincialis, in the presence or the absence of cadmium (Cd). TA at concentrations up to 20 μM, primarily found to be no toxic (in terms of cell viability, superoxide anions, nitric oxide and lipid peroxidation products currently estimated), significantly diminished the cytotoxic and oxidative effects induced by the metal (50 and/or 100 μM) in all cases. On the other hand, higher concentrations of TA (40 and 60 μM) were toxic, thus enhancing Cd-mediated cytotoxic and oxidative effects. The present study showed TA beneficiary properties in hemocytes of mussels, at least at low concentrations, while TA at concentrations higher than 20 μM could serve as an excellent oxidized substrate, thus enhancing toxic effects either alone or with the presence of micromolar concentrations of non transition metals, such as Cd.     

醚菊酯对日本囊对虾肝胰腺细胞DNA损伤效应的研究

为检测中国近海海洋环境中菊酯类农药等污染物对日本囊对虾(Marsupenaeus japonicas)的基因毒性及其危害程度,作者首次使用醚菊酯对日本囊对虾进行毒理实验,将日本囊对虾分为6组进行不同剂量的处理,采用彗星实验分析技术对处理培育25 d的日本囊对虾DNA损伤情况进行检测,并通过Comet Score 1.5分析软件对实验图谱进行拖尾率、尾长、彗尾DNA相对含量、尾距、Olive距等DNA损伤指标的统计分析,在对各指标分别建立一元回归方程的基础上,又通过SPSS软件进行判定分析,筛选影响作用更大的检测指标,建立了多参数的多元回归方程。实验结果发现,与空白对照组相比,日本囊对虾各实验处理组肝胰腺细胞DNA均有不同程度的损伤且各检测指标均有显著性差异(P0.01),各检测指标与处理浓度表现出良好的剂量效应关系,随着处理浓度递增,各检测指标都呈规律性的增长趋势。回归分析和相关系数表明:各检测指标与处理浓度具有高度的相关性,多项式回归方程(P0.05)和多元回归方程(P0.005)均有显著性的统计学意义,通过多元回归方程可以有效地推断醚菊酯处理浓度和处理时间。     

Shore exposure affects mussel population structure and mediates the effect of epibiotic algae on mussel survival in SW Ireland

On rocky shores, the relative importance of abiotic and biotic processes that regulate community structure are thought to vary with levels of shore exposure. This can lead to characteristic features found on sheltered and exposed shores. This study identified differences in the population structure of mussels on exposed and sheltered rocky shores on Atlantic coasts of south-west Ireland. Direct interactions between epibiotic algae and their host mussels were also examined to test if potential effects varied with shore exposure. Mussel beds on sheltered shores were less dense and comprised larger mussels with greater rates of individual survival and growth than those on exposed shores. The results of a field experiment showed that algal epibionts had a negative effect on mussel survival on sheltered shores but not on exposed shores. Surprisingly, the presence of algal epibionts had no effect on mussel growth on either shore type. These findings contrast with those of previous studies. The effects of shore exposure and algal epibionts on mussels may be species-specific and may interact with other factors across different regions. This study shows that predictions of effects of exposure on mussel populations and their epibionts should only be based on specific experimental evidence and cannot be generalised across regions.     

Contribution of Raman scattering to upward irradiance in the sea

Measurements of underwater irradiance revealed that the vertical attenuance in upward irradiance for wavelengths above 520 nm decreased with increasing depth, while the attenuance in the remaining wavelength region and also the attenuance in the downward irradiance in the whole wavelength range kept almost constant values. In this paper, it is suggested that the decrease in the attenuance for the upward irradiance above 520 nm can be ascribed to the Raman scattering of water molecules excited by the intense blue-green light in the downward irradiance. The pure water Raman scattering function at a scattering angle of 90° is measured and the results are used for the theoretical computation of upward irradiance generated by Raman scattering. Then, the difference between observed upward irradiance and the upward irradiance obtained by extrapolation from that in the shallow layers is computed under the assumption of constant irradiance attenuance. Since this difference is expected to represent the upward irradiance generated by Raman scattering, its value is compared with the upward irradiance due to Raman scattering obtained by theoretical computation. The similarity between the two upward irradiances so evaluated supports the view that Raman scattering makes a significant contribution to upward irradiance in the longer wavelength region.     

翡翠贻贝肉在冷冻冷藏中质量的变化

本文通过定期测定冷冻冷藏中翡翠贻贝肉的干耗率、过氧化值（ＰＯＶ）、挥发性盐基氮（ＴＶＢ－Ｎ）、ｐＨ值等理化指标,以探索翡翠贻贝肉在冷冻冷藏中质量的变化。结果表明,引起冷冻冷藏中贻贝肉蛋白质的冷冻变化和老化、烹调时口感发渣等的主要原因是水分的干耗和肉的严惩脱水。     

中国近海裸甲藻类分类研究

裸甲藻的分类鉴定工作是其生理生态学研究和赤潮研究的基础,为进一步完善我国裸甲藻分类体系,并为裸甲藻鉴定提供参考依据,收集了国家海洋局第一海洋研究所近年来在中国近海海域进行浮游植物调查所采集的水样与网样标本,针对裸甲藻目进行鉴定,初步鉴定出裸甲藻25种,其中在我国海域首次记录的有6种。以《中国海洋生物名录》收录的分类体系为基础,参照World Register of Marine Species的最新分类系统,将鉴定的25种裸甲藻分为5个科15个属。     

Palaeopressure reconstruction to explain observed natural hydraulic fractures in the Cleveland Basin

Natural fractures observed within the Lower Jurassic shales of the Cleveland Basin show evidence that pore pressure must have exceeded the lithostatic pressure in order to initiate horizontal fractures observed in cliff sections. Other field localities do not show horizontal fracturing, indicating lower pore pressures there. Deriving the burial history of the basin from outcrop, VR and heat-flow data gives values of sedimentation rates and periods of depositional hiatus which can be used to assess the porosity and pore pressure evolution within the shales. This gives us our estimate of overpressure caused by disequilibrium compaction alone, of 11 MPa, not sufficient to initiate horizontal fractures. However, as the thermal information shows us that temperatures were in excess of 95 °C, secondary overpressure mechanisms such as clay diagenesis and hydrocarbon generation occurred, contributing an extra 11 MPa of overpressure. The remaining 8.5 MPa of overpressure required to initiate horizontal fractures was caused by fluid expansion due to hydrocarbon generation and tectonic compression related to Alpine orogenic and Atlantic opening events. Where horizontal fractures are not present within the Lower Jurassic shales, overpressure was unable to build up as high due to proximity to the lateral draining of pressure within the Dogger Formation. The palaeopressure reconstruction techniques used within this study give a quick assessment of the pressure history of a basin and help to identify shales which may currently have enhanced permeability due to naturally-occurring hydraulic fractures.     

Effects of 17beta-estradiol exposure in the mussel Mytilus galloprovincialis

Mussels Mytilus galloprovincialis were exposed to different concentrations of estradiol (20, 200, and 2000 ng/l) in a semi-static regime (1-day dosing intervals) for up to 7 days in an attempt to see how mussels dealt with exogenous estrogenic compounds. Sex hormone levels were determined in whole tissue. Free-estradiol was only significantly elevated at the highest exposure dose (up to 10-fold). Most of the estradiol was in the tissues as fatty acid esters (> 78%), which sharply increased in a dose-dependent manner (from 4 ng/g in controls to 258 ng/g at the high exposure group). In contrast, neither free nor esterified testosterone levels showed significant differences between control and exposure groups. The results suggest the existence of mechanisms that allow mussels to maintain their hormonal status, and the important role that fatty acid esterification may play within those mechanisms. Synthesis and conjugation rates of estradiol were further investigated by measuring the activity of P450 aromatase, and palmitoyl-CoA:estradiol acyltransferase, in digestive gland microsomal fractions. Overall, the study contributes to the better knowledge of molluscan endocrinology, and defines new mechanisms of regulation of free steroid-levels in mussels.     

Relationships of rainfall,river flow,and salinity to faecal coliform levels in a mussel fishery

Faecal coliform levels were determined for sea water and shellfish flesh from farms of the green mussel, Perna canaliculus, in Pelorus Sound, Marlborough Sounds, New Zealand. Simple linear models were developed to relate logarithmically transformed data on the faecal coliform levels to measures of fresh water entering the mussel beds. These models support guidelines to restrict harvesting following periods of heavy rain in order to comply with shellfish sanitation standards. Alternative guidelines are suggested based on the volume of river water entering the sound or salinity.     

低盐度形成的微生物膜对厚壳贻贝稚贝附着的影响

为研究环境因子在厚壳贻贝(Mytilus coruscus)稚贝附着过程中的调控作用, 作者探讨了低盐度对微生物膜生物构成、群落结构的影响及所形成微生物膜对厚壳贻贝稚贝附着的影响。在实验室条件下, 研究微生物膜的日龄与干质量、细菌密度和硅藻密度、叶绿素a含量的关系及其对厚壳贻贝稚贝附着的影响。通过DGGE指纹图谱技术对微生物膜中的细菌群落结构多样性进行了分析。研究发现,盐度13和23时形成的微生物膜能有效促进厚壳贻贝稚贝的附着, 且盐度23、28d时稚贝附着率最高, 达到72%。相关性分析表明, 微生物膜的诱导活性与盐度、干质量、细菌密度、硅藻密度、日龄呈显著正相关性, 与叶绿素a无相关性。微生物膜的干质量、附着细菌密度及底栖硅藻密度明显随着日龄的增加而增加, 叶绿素a含量与微生物膜日龄无显著相关性。细菌群落在厚壳贻贝稚贝附着过程中发挥重要调控作用。     

Partitioning of copper among copper-binding proteins in the mussel Mytilus edulis exposed to soluble copper

Partitioning of copper among copper-binding proteins was evaluated in digestive glands of Mytilus edulis exposed to soluble copper. Groups of mussels were held in flow-through bioassay systems and exposed to 25 μg Cu liter^?1 for up to 21 weeks. At three-week intervals, groups of 25 mussels were removed and the digestive glands were analyzed for copper-binding proteins by gel-permeation chromatography and atomic absorption spectrometry.Chronic exposure to copper resulted in increased amounts of copper in the low molecular weight (LMW) protein fraction, which contains metallothioneins, and in the high molecular weight (HMW) protein fraction, which contains metalloenzymes. Concentrations of copper in the LMW protein fraction increased and then appeared to plateau with long exposure times, whereas those in the HMW protein fraction continued to increase with exposure time.     

Relationship between visible and heritable chromosome damage in trout cells and embryos

The disruption of chromosome segregation which is detected visually by the anaphase aberration (aa) test suggests that unequal amounts of DNA are distributed to daughter cells and possibly to subsequent cell generations. To investigate this possibility trout cell cultures and trout embryos (blastodisc) were exposed to benzo[a]pyrene (B[a]P) and the nitrosamide, MNNG, respectively. They were then examined by flow cytometry to determine if anaphase damage resulted in unequal DNA distribution to daughter cells. Both B[a]P and MNNG produced a significant increase (P < 0·01) in aa in cultured cells after 48 h exposure. These values returned to normal by 10 days in the absence of the genotoxic agents, except for the highest concentration (0·5 μg/ml MNNG), which showed only a 50% recovery by that time. Likewise, the coefficient of variation (CV) of nuclear DNA content of the cells also rose significantly after treatment and remained elevated for as long as 14 days following exposure. Experiments with rainbow trout embryos also showed a significant increase in both aa and CV following exposure to MNNG. Flow cytometric analysis of DNA content of trout cells and embryos following exposure to mutagens showed an unequal distribution of DNA that was transmissible through several cell generations. These findings indicate that visible anaphase aberrations could predict heritable genetic defects such as those associated with aneuploidy.     

Self-thinning, biodeposit production, and organic matter input to the bottom in mussel suspension culture

Future mussel suspension-culture methods may increasingly bypass standard population density adjustment on culture gear. This likely to exacerbate mussel fall-off. The potential extent of mussel fall-off is unknown. Here I use Dynamic Energy Budget theory and self-thinning theory to model the contribution of mussel fall-off to the total input of organic matter to the bottom. I focus on mussel populations grown on collector ropes and undergoing space-regulated self-thinning. Over a complete growth cycle, fall-off of mussel biomass was three times as high as the quantity harvested and accounted for 59% of total input of organic matter to the bottom. Biomass fall-off with collectors was 6-30 fold higher than with sleeving methods, as estimated from the literature. The relative contribution of mussel fall-off increased in a step-like fashion with initial population density, indicating that either mussels or biodeposits may dominate the organic input to the bottom, unless year-to-year variability in spat abundance is very high. The relative contribution of mussel fall-off did not increase appreciably with the intensity of second-set, but total mussel fall-off did. Assessment of each input type to the bottom is required for forecasting the consequences of a shift in culture methods, as they have different, spatially structured, effects on benthic environments.