DNA strand breaks in grass shrimp embryos exposed to highway runoff sediments and sediments with coal fly ash

Embryo production was reduced in female grass shrimp exposed to sediments with added coal fly ash and to sediments collected from an estuarine station containing high PAH concentrations due to its proximity to a highway storm drain. Grass shrimp embryos exposed to pore water from the high PAH and high metal sediments showed both reduced hatching and increases in DNA strand breaks (comet assay). Sediments with added coal fly ash had high concentrations of vanadium and selenium which may have contributed to effects similar to those observed with sediments with high PAH. The embryo pore water bioassay (hatching/DNA strand breaks) gave results comparable to those observed for reproduction effects (reduced embryo production/embryo hatching) with female grass shrimp exposed to whole sediment.     

Effects of genotoxic compounds on DNA and development of early and late grass shrimp embryo stages

Early and late developmental stages of grass shrimp embryos were exposed to different concentrations of two genotoxicants, 2-methyl-1,4-naphthoquinone (MNQ) and 4-nitroquinoline-N-oxide (NQO). DNA strand breaks were assessed by the comet assay while embryo development effects were determined by % of embryos hatching. Early embryo stage embryos were significantly more sensitive to genotoxicants than late stages. For example, all stage 4 embryos failed to hatch at 1 microM NQO while 95% of stage 8 hatched at this concentration. High DNA tail moments, which are a measure of the number of DNA strand breaks, were found in late stage embryos exposed to genotoxicants. Early stage embryo development was effected by low concentrations of genotoxicants but no changes were observed in DNA tail moments. We suggest that high DNA moments in late embryo stages reflect high DNA repair activity, while early stages may lack a fully developed DNA repair system.     

Interactive effects of UV, benzo[alpha] pyrene, and cadmium on DNA damage and repair in embryos of the grass shrimp Paleomonetes pugio

We examined the link between DNA strand breaks and hatching rates in grass shrimp, (Paleomonetes pugio), embryos exposed to 0.2 microM benzo[alpha] pyrene (BP), 5 microM cadmium (Cd) and 330 kJ/m(2) UV light, either alone or together. After exposure, embryos were transferred to clean seawater with or without 5 microM Cd. Hatching rates and DNA strand breaks (Comet Assay) were determined. DNA lesions caused by exposure to BP, UV light, or BP/cadmium were rapidly repaired and were not associated with any effects on hatching. Exposure to Cd after exposure to BP or UV did not affect embryological development or DNA repair. Exposure to BP/UV resulted in a high level of DNA lesions which were slowly repaired. Exposure to cadmium following BP/UV exposure inhibited hatching and DNA repair. Adducts formed during exposure to BP/UV exposure may be difficult to excise or may saturate the nucleotide excision repair system.     

Dietary transfer of fluoranthene from an estuarine oligochaete (Monopylephorus rubroniveus) to grass shrimp (Palaemonetes pugio): Influence of piperonyl butoxide

The objective of this study was to determine the potential for dietary transfer of sediment-associated fluoranthene from tubificid oligochaetes (Monopylephorus rubroniveus) to grass shrimp (Palaemonetes pugio). Grass shrimp, either in the presence or absence of sublethal waterborne concentrations of the metabolic inhibitor, piperonyl butoxide (PBO), were fed fluoranthene-dosed oligochaetes for 5-days. All grass shrimp bioaccumulated fluoranthene; however, bioaccumulation was 3X higher in the presence of PBO. Trophic transfer coefficients (TTCs) were 0.02 and 0.01 in the presence and absence of PBO, respectively. Following the 5-day accumulation period, shrimp in both treatments were allowed to depurate for 3 days. Depuration rates were significantly higher in PBO-exposed shrimp. These results demonstrated that sediment-associated fluoranthene can be transferred through the diet from oligochaetes to grass shrimp, and the presence of PBO enhanced fluoranthene bioaccumulation. However, the comparatively low TTCs suggest that biomagnification of fluoranthene in estuarine food webs is low.     

DNA strand breaks (comet assay) and embryo development effects in grass shrimp (Palaemonetes pugio) embryos after exposure to genotoxicants.

Grass shrimp embryos develop in egg sacs (stages 1-10) attached to the female for 14-20 days after which they 'hatch' from the egg sacs into a swimming zoea stage (stage 11). Until they emerge from the egg sacs, embryos depend on lipids and lipovitellin stored within the egg. The percent of embryos which hatch after exposure to toxicants relative to controls was the basis of an embryo development assay. Exposure of embryos to chromium(III) chloride, sodium chromate, mercuric chloride, and 2-methyl-1,2-naphthoquinone (MNQ) resulted in a reduced hatching rate. In addition to effects on embryo development, DNA strand damage tests were carried out on contaminant-exposed embryos, using the single-cell electrophoresis method often referred to as comet assay. Development of stage 4 embryos was more affected by MNQ exposure than stage 7 embryos. The hatching rates of stages 4 and 7 embryos exposed to MNQ (172 micrograms/l) were 0 and 90%, respectively. DNA strand damage, measured as DNA tail moments, were 3.4 and 4.4, respectively. Thus, exposure of an early embryo stage to MNQ prevented full embryo development while development of later embryo stages was not affected. It may be that the DNA repair systems are more efficient in later embryo stages than in early stages and thus DNA damaged in the early stages affects development.     

Comet assays to assess DNA damage and repair in grass shrimp embryos exposed to phototoxicants

Exposure of grass shrimp (Palaemonetes pugio) embryos to four compounds (anthracene, pyrene, alpha-terthienyl, methylene blue) along with solar exposure resulted in extensive DNA strand damage using the comet assay. DNA tail moments of embryos exposed to these chemicals in the dark ranged from 1.8 to 4.3, while exposure to chemicals and solar resulted in tail moments of 14.3-15.3. Reduction of DNA tail moments when solar exposed embryos were transferred to the dark, suggested DNA repair systems were active. The comet assay can be used to follow both DNA damage and repair following exposure to phototoxic chemicals.     

Multi-level assessment of chronic toxicity of estuarine sediments with the amphipod Gammarus locusta: I. Biochemical endpoints

We report on biomarker responses conducted as part of a multi-level assessment of the chronic toxicity of estuarine sediments to the amphipod Gammarus locusta. A companion article accounts for organism and population-level effects. Five moderately contaminated sediments from two Portuguese estuaries, Sado and Tagus, were assessed. Three of them were muddy and two were sandy sediments. The objective was to assess sediments that were not acutely toxic. Three of the sediments met this criterion, the other two were diluted (50% and 75%) with clean sediment until acute toxicity was absent. Following 28-d exposures, the amphipods were analysed for whole-body metal bioaccumulation, metallothionein induction (MT), DNA strand breakage (SB) and lipid peroxidation (LP). Two of the muddy sediments did not cause chronic toxicity. These findings were consistent with responses at organism and population levels that showed higher growth rates and improvement of reproductive traits for amphipods exposed to these two sediments. Two other sediments, one muddy and one sandy, exhibited pronounced chronic toxicity, affecting SB, MT induction (in muddy sediment), survival and reproduction. Potential toxicants involved in these effects were identified. The last sandy sediment exhibited some loss of DNA integrity, however growth was also enhanced. Present results, together with the organism/population-level data, and also benthic communities information, were analysed under a weight-of-evidence approach. By providing evidence of exposure (or lack of it) to contaminants in sediments, the biomarkers here applied assisted in distinguishing toxicants' impacts in test organisms from the confounding influence of other geochemical features of the sediments.     

Different levels of mussel (Mytilus edulis) DNA strand breaks following chronic field and acute laboratory exposure to polycyclic aromatic hydrocarbons

Levels of polycyclic aromatic hydrocarbons (PAHs) including benzo[a]pyrene (B[a]P) were at least seven-fold higher in mussels sampled from a polluted site (Loch Leven, in Scotland, UK) compared to a nearby clean reference site (Loch Etive) throughout the year 2000. Levels of DNA strand breaks (alkaline COMET assay) using both gill and digestive gland nuclei were similar at both sites despite the difference in contaminant load (total PAH). In contrast, mussels collected from a reference site (Port Quin, Cornwall, UK) had an increase in DNA strand breaks in digestive gland cells following laboratory exposure to B[a]P-dosed Isochrysis galbana. However, after 14 days high dose (20 ppb-exposed diet) animals had returned to levels similar to the controls. There was no evidence of increased necrosis or apoptosis after treatments. The results from these two studies suggest that an adaptive response may prevent ongoing DNA damage in mussels exposed to high levels of B[a]P and PAH contamination.     

Biomass of meiobenthic in the Bohai Sea, China

IorasMeiOfauna is a vmp imprtant pep in benthic small bo web energetica1ly duo to theirarell aize and high turnovr rates. A nde Of meMauna op be the recireulaton of nutrient8*Marine nemathe mey keep the bacterial chnies on sand grains in active phase Of gorth byfeding on bacteria, thus enhancing the recirculation Of nutrients (McIntyre, l969; Feller andWrmick, l988; MOntagna et al., l995). But energetics studie8, no matter tfor conductedon the energybudgt level for individua sPeCies or a…     

Measurement of DNA single-strand breaks in gill and hemolymph cells of mussels, Mytilus sp., collected on the French Atlantic Coast

DNA single-strand breaks were measured by the comet assay in both gill and hemolymph cells of mussels collected in 3 sampling areas of the French coast (Pointe du Castelli, Pen Bron and Saint-Nazaire Harbour). Whole mussel tissue samples were also collected for the chemical determination of PAH, PCB and heavy metal concentrations. In mussel, a higher level of DNA strand breaks was measured in gill than in hemolymph cells (p < 0.01). Despite a factor of contamination from 2 to 3 between sites, no difference in the extent of mussel DNA strand breaks was shown between sampling locations (p > 0.05), questioning the sensitivity of the assays used in biomonitoring studies.     

The structure of the benthic macrofaunal assemblages and sediments characteristics of the Paraguaçu estuarine system, NE, Brazil

The structure of the benthic macrofaunal assemblages of the estuarine portion of Paraguaçu River, NE, Brazil, and its relationship with surface sediment characteristics (trace metals, PAHs, nutrients and grain size) and physical variables were investigated at ten stations on two contrasting occasions, summer (dry season) and winter (rainy season). A total of 1258 individuals (632 in winter and 626 in summer) and 62 taxa representing polychaetes, crustaceans, bivalves, echinoderms, bryozoans, sponges, cnidarians and cephalochordates were collected. Benthic assemblages in the upper estuary were unlike those in the lower estuary and a clear substitution of benthic taxa along the estuary was observed. Macrofaunal invertebrates in the low salinity region, composed of coarse sediments, were dominated by tellinids, venerids (bivalves), cirolanids (isopods), cyclopoids (copepods), and nereidids (polychaetes). While the high salinity region, composed of fine sediments, were dominated by nuculids (bivalves), cirratulids (polychaetes), and by amphiurids (ophiuroids). The Paraguaçu estuarine system is not severely affected by anthropogenic activities. In the great majority of the study sites, concentrations of trace metals and PAHs in the sediments were near background values. Nutrients values were also low. We formulated new models of taxon distribution and suggested detailed studies on the effects of salinity variation and studies using functional approaches to better understand the processes causing the spatial patterns in tropical estuarine benthic assemblages.     

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 amino acid composition of estuarine colloidal material

Amino acids were determined in estuarine colloidal and particulate material from near surface waters ranging from fresh to brackish water (12 g kg^?1 salinity). The hydrolizable amino acids and associated ammonia account for an average of 80% of the nitrogen present in colloidal samples, and approximately 75% of the nitrogen from particulate fractions of the same samples. The relative proportions of these amino acids are similar to those of a cultured estuarine diatom. There are no significant amounts of D amino acids, or non-protein amino acids characteristic of bacteria in sediments and soil. A single deep-water sample (25 m) shows some evidence of bottom mud resuspension by the presence of a possible aspartic acid-hydroxyproline dimer.     

Fatty acids and sterols as source markers of organic matter in sediments of the North Carolina continental slope

To estimate the source and diagenetic state of organic matter reaching bottom sediments, fatty acids and sterols were measured in unconsolidated surface material (flocs) at 12 sites ranging from 600 to 2000 m across the mid-Atlantic continental slope off Cape Hatteras, North Carolina. Total free and esterefied fatty acids were similar in distribution and concentration to other coastal systems, with values ranging from 0.64 to 46.52 μg mg⁻¹ organic carbon (1.10–68.85 μg g⁻¹ dry sediment). Although shallow (600 m) stations contained significantly greater fatty acid concentrations than deep (> 1400m) stations, high variability observed at mid-depth (800 m) collections precluded a consistent relationship between total fatty acid concentration and station depth. At three sites where underlying sediments were also collected, decreases in total fatty acids, reduced amounts of polyenoic acids and significant presence of bacterial fatty acid suggest rapid reworking of labile organic material that reaches the sediment surface. The distribution of sterols was remarkably consistent among all sites even though there were large variations in concentrations (1.8–20.7 μg mg⁻¹ organic carbon). Sterol composition indicated phytoplankton, principally diatoms and dinoflagellates, as the principal source of labile organic matter to sediments, together with a significant input of cholest-5-en-3β-ol typical of zooplankton and their feeding activity. A minor but widespread terrigenous input was also evident based upon significant concentrations of sterols dominant in vascular plants.     

Temporal Variations of Benthic Bacterial Microflora on the Northwestern Mediterranean Continental Shelf and Slope

Abstract. Bacterial (direct and heterotrophic counts, heterotrophic potentials) and organic matter (org C, org N) parameters were studied over 2 years in the surface sediments of 3 stations located on the continental shelf (station A: 15m; station B: 35m) and slope (station C: 910m) in the northwestern Mediterranean Basin.
Although logistic constraints did not allow a satisfactory sampling periodicity in the offshore survey, some seasonal trends were still observable at the deepest station. Variations were quantitatively more important in the two shallow sediments (from 3×10² heterotrophic bacteria – ml^-1 in December 1984 to 10⁷ heterotrophic bacteria-ml^-1 in March 1986). Epifluorcsccncc direct counts were relatively constant (5×10⁸ cells-ml^-1), while heterotrophic potentials (with ¹⁴C labelled glucose and glutamic acid) showed a succession of very short pulses over the year. Different concentrations of organic matter (from 0.2% orgC at 15m to 0.8% orgC at 35m) appeared to sustain very similar bacterial numbers. Nevertheless, a certain relation between the seasonal evolution of bacterial and organic matter parameters was discernable. Although depth apparently had no measurable effect on maximal spring bacterial densities, it seemed to lessen the summer and winter decreases in the two shallow stations.     

近海水体环境DNA沉降对沉积物中纤毛虫分子多样性评估的影响

在陆架海区沉积物中, DNA高通量测序技术可检获大量浮游寡毛类和舞毛类等非底栖纤毛虫,这些源于水体的环境DNA(eDNA)如何影响对沉积物中纤毛虫分子多样性的评估,以及影响程度如何尚不明确。本研究选取了黄海冷水团中的两个站位,通过提取水体和沉积物中DNA和RNA,并采用直接提取法和洗脱法提取沉积物DNA,结合DNA和cDNA测序技术,探讨了水体和沉积物中纤毛虫分子多样性的关系。研究表明,基于洗脱DNA法、直接提取DNA法和cDNA法获得的沉积物中纤毛虫OTUs数分别为451、312和324个,其中211个OTUs同时由三种方法检获;而164个OTUs仅通过洗脱DNA法检获,其中89%为相对丰度低于0.1%的稀有类群。直接提取DNA法所获的寡毛类和舞毛类序列数占比达46%,而在洗脱DNA法和cDNA法中占比仅为12%和10%。沉积物中检获的43%—71%的舞毛类和寡毛类OTUs与浅层水(40m)共有,仅19%—29%与深层水(40m)共有,且这些共有的OTUs可同时在浅层水中检获。本研究发现,对沉积物中纤毛虫分子多样性评价造成影响的浮游类群主要来自浅层水,洗脱DNA与cDNA测序均可显著降低浮游类群eDNA的影响。鉴于洗脱DNA法较之cDNA法操作更简便,且避免了反转录过程导致的偏差,因此推荐用于对近海沉积物中纤毛虫等真核微生物的分子多样性研究。     

环境因子对菲在黄河口沉积物上吸附的影响

为研究多环芳烃从河口到近海的环境归趋行为与生态风险,考察了沉积物质量浓度、溶解性有机质、温度、盐度4种典型环境因子对菲在黄河口沉积物上吸附的影响,比较了黄河口与近海两种沉积物对菲的吸附性能。研究结果表明,沉积物质量浓度越低,单位质量颗粒物的菲吸附量越高;共存的溶解性有机质对菲的吸附具有增促作用,且腐殖酸比黄腐酸的作用更显著;温度的升高不利于菲的吸附,而盐度的增加有利于菲的吸附。菲在沉积物上的吸附是分配作用与表面吸附两种行为的耦合,其中黄河口沉积物以表面吸附为主,而近海沉积物以分配作用为主。近海沉积物菲吸附量显著高于黄河口沉积物菲吸附量。基于此,菲从河口到近海的迁移过程中,更易于在沉积物表面发生吸附沉降,从而可能降低水相中的生态危害,但对近海底栖生物具有潜在的健康生态风险。     

长江口及毗邻海区沉积物中磷的分布特征

于2006年6月和2007年4月在长江口及毗邻海区采集了表层沉积物样品,利用水淘选法对沉积物进行了粒径分级,并采用改进后的SEDEX方法对沉积物样品及分粒级沉积物样品进行了磷形态分析。结果表明,2006年6月和2007年4月长江口及毗邻海区沉积物总磷变化分别为12.56~19.64μmol/g和8.99~19.91μmol/g,其中碎屑磷是磷的主要存在形态。可交换态磷、铁结合态磷、有机磷、自生磷灰石磷及难分解有机磷在长江口外的平面分布比较一致,均在紧邻长江口门外、杭州湾外的条状带出现了含量的高值,而在口门内及口门外河口靠海一侧碎屑磷含量较高。不同形态磷在不同粒径沉积物中的含量不尽相同:有机磷、自生磷灰石磷以及难分解有机磷是小于8μm粒级沉积物中磷的主要组成部分,而且其在总磷中所占比例随沉积物粒径的增加而下降;碎屑磷则主要集中在粗沉积物中,为32~63μm及大于63μm粒级沉积物中磷的主要贡献者。沉积物的粒度分异以及各形态磷在不同粒径沉积物中含量的差异,共同影响着河口沉积物中磷的分布。     

The use of free-living nematodes as a bioassay for estuarine sediments

A bioassay technique using free-living nematodes has been developed to assess the environmental quality of estuarine sediments. Nematodes offer the advantages of (a) generally being the most abundant metazoan in sediments; (b) having short life histories and no pelagic larvae; and (c) being relatively easy to maintain in laboratory culture. Small individual body size and rapid generation time (two weeks) render nematodes especially useful in evaluating effects of potential toxicants on populations, rather than on single individuals.The bioassay technique was developed using two species, Chromadorina germanisa and Diplolaimella punicea, and sediments from the Hudson-Raritan estuarine area. The life histories and feeding habits of both species are known. Test sediments were taken from eight stations that represent a gradient from lightly to heavily impacted. Concentrations of potential toxicants measured in these sediments included polychlorinated hydrocarbons, polynuclear aromatic hydrocarbons, and heavy metals. Control sediments, taken from 100 km distant, were almost totally toxicant-free. Nematodes were incubated in the dark at 25°C for two weeks at 1:1 and 1:10 concentrations (volume/volume) of sediments prepared with sterile sea water. Food consisted of bacteria from nematode stock cultures. Intrinsic rate of natural increase (r) of the nematodes was used as the indicator of sediment quality; r has units (daily increase in number of generations), is easily computed, and can be treated statistically.Both species grew better in control and other sediments containing low concentrations of PCBs, PAHs, and heavy metals. In sediments having TIF PCB and PAH concentrations (ppb) of more than 270 and 8700, respectively, population growth of both species tended to be less than half that found at lower concentrations. Significant inverse correlations between r and toxicant concentration occurred at 1:1 sediment-water concentrations. At 1:10 concentrations the correlations remained negative, but not at the 0·05 level of significance.Use of the nematodes Chromadorina germanica and Diplolaimella punicea offers a quick, easy and statistically testable means of assessing the quality of marine sediments on populations, rather than single individuals, of marine benthic invertebrates.     

Behaviour of proteinaceous substances in the estuary of the Tama River

Behaviour of dissolved and particulate proteinaceous substances in the estuary of the Tama River was investigated through field observations and laboratory experiments. Concentrations of dissolved proteinaceous substances in estuarine water were determined at eight stations between the mouth of the Tama River (TA-15) and a point 16km upstream (TA-8), using the colorimetric method of Lowryet al. (1951) that employs Folin phenol reagent. The results were expressed in terms of Folin phenol active substances (FPAS). It was found that FPAS decreased with increasing salinity and that the proportion of high molecular weight FPAS (molecular weight greater than 100,000) decreased rapidly in the estuary.The shapes, sizes and numbers of amorphous particulate substances stained by Amido Black 10B, called Amido Black active substances (ABAS), were determined under a microscope. The numbers of ABAS retained on Millipore HA filters (mean diameter: 10–80µm) amounted to 450ml^–1 in the estuarine water about 6km from the mouth of the estuary (Station TA-12M). Some of these aggregates are considered to be producedin situ by flocculation of high molecular weight FPAS during estuarine mixing. ABAS were also produced in experiments by mixing filtered river water with sea water.