Sperm motility in the Pacific oyster (Crassostrea gigas) is affected by nonylphenol

Nonylphenol, a widespread aquatic pollutant with endocrine-disrupting potential has been found to affect sperm motility in the Pacific oyster, Crassostrea gigas (Thunberg). This marine invertebrate, a global aquaculture species, was exposed to nonylphenol at concentrations of 1 and 100 μg l⁻¹ for a duration of 72 h during the period of gametogenesis. Growth was monitored at regular intervals until sexual maturity when sperm motility was assessed. The growth rate of C. gigas remained unaffected by exposure to nonylphenol during gametogenesis. However, the number of individuals with motile sperm was significantly reduced.     

Effects of heavy metals on the survival and feeding behaviour of the sandy shore scavenging gastropod Nassarius festivus (Powys)

The effects of Cu, Cd, Zn and Cr on the survival and feeding behaviour of the sandy shore scavenging gastropod Nassarius festivus were compared. The 96-h LC₅₀ for Cu, Cd, Zn and Cr were 0.36, 1.52, 1.76 and 36.9 mg l⁻¹, respectively. Four sublethal concentrations of each metal plus a control were prepared and the snails were exposed to experimental solutions for 96 h. Feeding behaviour was studied after the snails were starved for five days. As compared with the control, the number of individuals feeding was significantly reduced by exposure to 0.05 mg l⁻¹ Cu, 0.2 mg l⁻¹ Zn, 0.5 mg l⁻¹ Cd and 5 mg l⁻¹ Cr. The time spent feeding was greater for individuals exposed to greater concentrations of Zn and Cd but no effect was found for Cu and Cr. Chemoreception of food was studied by placing the snails at a fixed distance of 15 cm from the bait. The success rate of reaching the bait was less for individuals exposed to Cr but no effect was found for Zn, Cu or Cd. The time required for an individual to reach the bait decreased as the concentration of Zn increased. In contrast, a longer time was required for individuals exposed to Cr whereas the effect of Cd and Cu was insignificant. The potential of using feeding behaviour and chemoreception in contaminant evaluations is discussed.     

Burial, crawling and drifting behaviour of the bivalve Macomona liliana in response to common sediment contaminants

To investigate the effect of sediment contamination on the tellinid bivalve Macomona liliana, juvenile shellfish (about 1.3 mm long) were exposed to copper- and zinc-dosed sand, and sediments collected from several contaminated sites. The behavioural responses measured included burial rate, crawling and drifting. In copper-dosed sediment, a significant reduction in the number of shellfish burying after 10 min occurred at 25 mg Cu kg⁻¹ (dry wt). After a 96-h exposure, shellfish crawled away from sediment with 10 mg Cu kg⁻¹ (dry wt), and when a weak current was provided they left this 10 mg Cu kg⁻¹ (dry wt) sediment by drifting. Zinc-dosed sediment slowed burial at 80 mg Zn kg⁻¹ (dry wt), and stimulated crawling and drifting at 40 mg Zn kg⁻¹ (dry wt). Some of the field-collected contaminated sediments slowed burial and stimulated drifting, but none affected crawling. It appears that juvenile drifting induced by sediment contamination could play a role in determining M. liliana distributions.     

Biomonitoring of pollution by Cerastoderma edule from the British Isles: a Laser Ablation ICP-MS study

Shells of the common cockle Cerastoderma edule have been analysed using Laser Ablation Inductively Coupled Plasma-Mass Spectrometry (LA-ICP-MS) for a suite of elements (Pb, Cu, As, Zn and U). The samples of C. edule were obtained from four sites in the British Isles, two facing the Atlantic and two adjacent to the Irish Sea. The LA-ICP-MS technique, permits high resolution examination of the relationship between the concentrations of elements in the hard parts and that in the environment. This study shows that this bivalve can be used as a biomonitor of pollution. All four areas under investigation are affected by anthropogenic pollution, in particular Zn and Cu, and are characterized by short-term extreme pollution events. Furthermore, high levels of U are recorded from the cockles sampled from the sites adjacent to the Irish Sea, probably related to a combination of both natural and industrial effluents discharged into the area.     

Effects of Animal Size on Concentrations of Metallothionein and Metals in Periwinkles Littorina littorea Collected from the Firth of Clyde, Scotland

Different sizes of Littorina littorea were collected from four areas in the Firth of Clyde, Scotland. Their metallothionein (MT) and heavy metal concentrations were analysed using the Silver Saturation Method and Atomic Absorption Spectrophotometry, respectively. Concentrations of MT, Cd and Zn (as μg g⁻¹ dry soft-body weight) generally decreased with an increase in size of L. littorea. MT concentrations were better correlated with Cd than with Zn or Cu concentrations. Nevertheless, MT and the metals in periwinkles (as μg individual⁻¹) increased significantly with increasing size. Concentrations of MT and the metals among the sampling areas were compared at a standardised soft-body weight (10 mg). The results from Discriminant Analysis based on all metal concentrations indicate that Largs is different from the other areas and characterised by high Fe concentrations in L. littorea. The problems and differences in using either soft-body weight or shell length as independent variable for size-standardisation are discussed.     

The potential role of mucus in the depuration of copper from the mussels Perna viridis (L.) and Septifer virgatus (Wiegmann)

Experiments were conducted to measure the mucus secretion rate in two common mussels in Hong Kong, Perna viridis (L.) and Septifer virgatus (Wiegmann), when chronically exposed to Cu (50 μg l⁻¹). After a 3-month exposure period, the mucus production rate of P. viridis at 25°C in the metal treatment was 2.65 times that of the control (10.7 vs 4.0 mg g⁻¹ dry wt h⁻¹), while S. virgatus showed a 1.85 times difference (4.41 vs 2.38 mg g⁻¹ dry wt h⁻¹). Mucus secretion by P. viridis under acute Cu exposure (0.5 mg l⁻¹) was significantly higher in the metal treatment than the control (13.43 vs 9.16 mg g⁻¹ dry flesh wt). Metal contents of the mucus secreted was about 18 times those in the control and 6 times in the soft tissues. Therefore, mucus appears to be an effective agent for Cu depuration in the mussel. The significance of these results to the local distribution and abundance of the mussels is discussed.     

Physiological Responses of Five Seagrass Species to Trace Metals

Trace metal run-off associated with urban and industrial development poses potential threats to seagrasses in adjacent coastal ecosystems. Seagrass from the largest urban (Moreton Bay) and industrial (Port Curtis) coastal regions in Queensland, Australia were assessed for metal concentrations of iron (Fe), aluminium (Al), zinc (Zn), chromium (Cr) and copper (Cu). Trace metal concentrations in seagrass (Zostera capricorni) leaf and root-rhizome tissue had the following overall trend: [Fe] > [Al] > [Zn] > [Cr] > [Cu]. Rainfall events and anthropogenic disturbances appeared to influence metal concentrations in seagrasses with the exception of Al, which does not appear to bioaccumulate. In laboratory experiments, five seagrass species (Halophila ovalis, H. spinulosa, Halodule uninervis, Z. capricorni, Cymodocea serrulata) were incubated with iron (1 mg Fe l⁻¹) and copper (1 mg Cu l⁻¹) and responses assessed by changes in PSII photochemical efficiency (Fv/Fm), free amino acid content and leaf/root-rhizome metal accumulation. Iron addition experiments only affected Halophila spp, while copper additions affected other seagrass species as well. Trace metal contamination of seagrasses could have ramifications for associated trophic assemblages through metal transfer and seagrass loss. The use of photosystem II photochemical efficiency as well as amino acid concentrations and composition proved to be useful sublethal indicators of trace metal toxicity in seagrasses.     

Calcium isotope fractionation in coccoliths of cultured Calcidiscus leptoporus, Helicosphaera carteri, Syracosphaera pulchra and Umbilicosphaera foliosa

Four species of marine calcifying algae, the coccolithophores Calcidiscus leptoporus, Helicosphaera carteri, Syracosphaera pulchra and Umbilicosphaera foliosa were grown in laboratory cultures under temperatures varying between 14 and 23 °C, and one species, C. leptoporus, under varying [CO₃²⁻], ranging from 105 to 219 μmol/kg. Calcium isotope compositions of the coccoliths resemble in both absolute fractionation and temperature sensitivity previous calibrations of marine calcifying species e.g. Emiliania huxleyi (coccolithophores) and Orbulina universa (planktonic foraminifera) as well as inorganically precipitated CaCO₃, but also reveal small species specific differences. In contrast to inorganically precipitated calcite, but similar to E. huxleyi and O. universa, the carbonate ion concentration of the medium has no statistically significant influence on the Ca isotope fractionation of C. leptoporus coccoliths; however, combined data of E. huxleyi and C. leptoporus indicate that the observed trends might be related to changes of the calcite saturation state of the medium. Since coccoliths constitute a significant portion of the global oceanic CaCO₃ export production, the Ca isotope fractionation in these biogenic structures is important for defining the isotopic composition of the Ca sink of the ocean, one of the key parameters for modelling changes to the marine Ca budget over time. For the present ocean our results are in general agreement with the previously postulated and applied mean value of the oceanic Ca sink (Δ_sed) of about − 1.3‰, but the observed inter- and intra-species differences point to possible changes in Δ_sed through earth history, due to changing physico-chemical conditions of the ocean and shifts in floral and faunal assemblages.     

The Impact of the Herbicide Diuron on Photosynthesis in Three Species of Tropical Seagrass

The impact and recovery from exposure to the herbicide diuron [DCMU; 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea] was assessed for three tropical seagrasses, maintained in outdoor aquaria over a 10-day period. Photosynthetic stress was detected using chlorophyll a fluorescence, measured with a Diving-PAM (pulse amplitude modulated fluorometer). Exposure to 10 and 100 μg l⁻¹ diuron resulted in a decline in effective quantum yield (ΔF/F_m^′) within 2 h of herbicide exposure in Cymodocea serrulata, Halophila ovalis and Zostera capricorni. Effective quantum yield also declined over the first 24 h of exposure in H. ovalis at even lower diuron concentrations (0.1 and 1.0 μg l⁻¹). Effective quantum yield in H. ovalis and Z. capricorni was significantly depressed at all diuron concentrations (0.1–100 μg l⁻¹) after 5 days exposure, whereas effective quantum yield in C. serrulata was only significantly lower in plants exposed to highest diuron concentrations (10 and 100 μg l⁻¹). Effective quantum yield depression was present 5 days after plants exposed to 10 and 100 μg l⁻¹ diuron were returned to fresh seawater. These results indicate that exposure to herbicide concentrations present in nearshore Queensland sediments present a potential risk to seagrass functioning.     

Calibrations for benthic foraminiferal Mg/Ca paleothermometry and the carbonate ion hypothesis

Benthic foraminiferal magnesium/calcium ratios were determined on one hundred and forty core-top samples from the Atlantic Ocean, the Norwegian Sea, the Indian Ocean, the Arabian Sea and the Pacific Ocean, mostly at sites with bottom water temperatures below 5 °C. Mg/Ca ratios are consistently lower, by  0.2 mmol/mol, in samples cleaned using oxidative and reductive steps than using oxidative cleaning. Differences between Cibicidoides species have been identified: Mg/Ca of Cibicidoides robertsonianus > Cibicidoides kullenbergi > Cibicidoides wuellerstorfi. Comparison with bottom water temperatures support observations of lowered Mg/Ca of C. wuellerstorfi at temperature below  3 °C compared with values predicted by published calibrations and from other Cibicidoides species. Hydrographic data shows that carbonate ion saturation (Δ[CO₃²⁻]) decreases rapidly below this temperature. An empirical sensitivity of Δ[CO₃²⁻] on Mg/Ca has been established for C. wuellerstorfi of 0.0086 ± 0.0006 mmol/mol/μmol/kg. A novel application using modern temperatures and Last Glacial Maximum temperatures derived via pore fluid modelling supports a carbonate ion saturation state effect on Mg incorporation. This may significantly affect calculated δ¹⁸O_seawater obtained from foraminiferal δ¹⁸O and Mg/Ca temperature.     

Sublethal effects of cupric ion activity on the grazing behaviour of three calanoid copepods

This research (1) characterized the effects of sublethal cupric ion activities on the grazing behavior of two estuarine copepods (Acartia tonsa, Acartia hudsonica) and one nearshore, neritic copepod (Temora longicornis) and (2) compared the sensitivity of short-term sublethal behavioral assays with that of longer-term acute toxicity tests. A nitrilotriacetate-trace-metal-ion buffer system at 27‰. S was used to quantify and control the free cupric ion activity. Acute toxicity tests were used to determine the mortality of A. tonsa and T. longicornis over 72 h within the approximate cupric ion activity range of 10⁻¹³ to 10^−9.5 M. 24 h survival was not affected within the approximate cupric ion activity range of 10⁻¹³ to 10^−9.7 M, the range used for subsequent grazing activity experiments after 24 h exposure to Cu. Grazing activity was significantly diminished at cupric ion activities of ≈ 10⁻¹⁰ M for A. tonsa and T. longicornis, and at ≈ 10⁻¹¹ M for A. hudsonica. A hormetic pattern of response in feeding activity was observed with A. tonsa and T. longicornis. Grazing activity was found to be a sensitive measure of sublethal Cu stress compared with the acute toxicity tests. Grazing activity was affected at environmentally relevant cupric ion activities.     

Taxonomic composition and seasonality of diatoms in three Dinaric karstic lakes in Croatia

The composition and temporal distribution of planktonic diatoms were studied in three small shallow karstic lakes – Modro Oko, Desne, and Kuti – throughout the seasons in 1994, 1995, 1997, and 1998. Physico-chemical parameters did not differ among the three lakes, except for NO₃ concentrations. Altogether, 137 diatom taxa within 41 genera were found in all three lakes combined. The highest number of taxa belonged to the genera Surirella (13), Navicula (12), andCymbella (9). Most taxa were tychoplanktonic, with true panktonic taxa contributing only 14% of the total. Despite their physical-chemical similarity, the lakes showed differences in the structure and dynamics of their diatom communities. Blooms occurred at different times in each of the lakes, and community similarity (Jaccard index) among all three was low (17.6–29.1%). The highest mean values of surface diatom abundance in Modro Oko (2199 cells L⁻¹), Desne (7900 cells L⁻¹), and Kuti (20690 cells L⁻¹) were not significantly different. Margalef's species richness index did not differ either among the lakes or seasonally. Only five diatom taxa in quantitative samples, and 22 taxa in net-samples, were common to all lakes. The most frequent diatoms were Cyclotella striata (56–93%), Fragilaria ulna (50–67%), and Asterionella formosa (27–83%). The most abundant taxa in Modro Oko, Desne, and Kuti were, respectively, Cymbella sp. 1 (402 cells L⁻¹), Cyclotella striata (3000 cells L⁻¹) and Asterionella formosa (20690 cells L⁻¹). Eighteen taxa were recorded for the first time in Croatia: Amphora commutata, Caloneis amphisbaena var. subsalina, C. silicula var. peisonis, Campylodiscus echeneis, Coscinodiscus lacustris, Denticula elegans, Gyrosigma acuminatum var. lacustre, G. distortum var. parkeri, G. fasciola, Navicula menisculus var. menisculus, N. pusilla, N. helensis, Stauroneis parvula var. prominula, Staurosira construens f. subsalina, Stephanodiscus sp. 1, Surirella gracilis, S. ovalis, and S. striatula. The genus Stephanodiscus was recorded for the first time in Croatia.     

Clam burrowing behaviour: Inhibition by copper-enriched sediment

Burrowing behaviour is adaptive and allows clams to escape predation; yet the effects of potentially toxic metals on such behaviour have not been adequately investigated. In natural marine sediment contaminated with copper the time for littleneck clams (Protothaca staminea) to achieve complete burial was recorded. Above a threshold of 5.8 μg g⁻¹ Cu added to dry sediment, the time for 50% of the clams to burrow (ET₅₀) increased logarithmically with increasing sediment copper concentration according to:

logET₅₀ = 0.15 (CU) - 1.37 (n = 4, r² = 0.98)

where ET₅₀=time in hours for 50% of clams to burrow and Cu=μg g⁻¹ Cu in dry sediment. Previously exposed clams had both a lower threshold to Cu and a longer reburrowing time (ET₅₀). Clams exposed to sediment mixed with Chelex-100®-sorbed copper showed no significant change in burrowing time. Bioassays based on claim burrowing behaviour can measure both bioeffectiveness of sediment-sorbed metals and a sublethal effect with ecological meaning.     

Biomonitoring of heavy metals and trace organics using the intertidal mussel Perna viridis in Hong Kong coastal waters

This paper presents the results of a 6-year (1998–2003) survey of trace toxics in the intertidal mussel Perna viridis conducted by the Hong Kong Environmental Protection Department. Concentrations of heavy metals and trace organics were measured in the soft bodies of P. viridis collected from five sites in Hong Kong waters, i.e. Wu Kai Sha (Tolo Harbour), Ma Wan (Northwest), Tsim Sha Tsui (Victoria Harbour), Tai Tam (Hong Kong South) and Lamma Island (Southern Waters) in order to establish the spatial patterns of contaminants in mussels. Among the metals analysed, Cd showed a significant concentration gradient in Hong Kong waters. The levels of Cd in P. viridis were significantly higher at Ma Wan as compared to the other sites studied. Ma Wan also had relatively higher concentrations of Pb. Mn concentrations were particularly prominent at Wu Kai Sha. Significantly higher concentrations of Hg and Cu were recorded at Tai Tam and Tsim Sha Tsui. Tai Tam and Wu Kai Sha had higher levels of V; whereas higher Ni concentrations were recorded at Lamma Island and Tai Tam. No clear spatial patterns for Al, As, Cr, Fe and Zn were observed.

Higher concentrations of PAHs in P. viridis were observed around urban centres impacted by sewage discharges (e.g. Tsim Sha Tsui); whereas higher PCB levels were found not only in Tsim Sha Tsui but also in less urbanised areas such as Lamma Island and Tai Tam, suggesting that these may be due to non-sewage related inputs. The study also shows that Northwest and Southern waters are subject to a higher degree of DDT pollution compared with other sites. Of the 17 dioxin compounds analysed, positive data were mostly recorded for two compounds which are of low toxicity (i.e. OCDD and 1,2,3,4,6,7,8-HpCDD) whereas the most toxic congeners (i.e. 2,3,7,8-TCDD and 1,2,3,7,8-PeCDD) were not detected in the 6 years of monitoring. In general, the levels of OCDD in P. viridis were found to be higher in Tai Tam and Lamma Island in Southern Waters of Hong Kong.

This study found that the levels of some highly toxic heavy metals (i.e. Cd, Hg and Pb) in the mussel P. viridis did not exceed the recommended limits for shellfish as food in Hong Kong (i.e. Cd: 2.0 ppm; Hg: 0.5 ppm; Pb: 6.0 ppm wet weight). The levels of As and Ni in P. viridis were also well below the action limits set by the US FDA (i.e. As: 86 ppm; Ni: 80 ppm wet weight). DDT and PCB contaminations in P. viridis were below the concentrations of concern. Compared with data obtained in the 1980s, the current levels of DDTs in P. viridis were 4–16 times lower; whereas Pb concentrations recorded in Tsim Sha Tsui have also been lowered significantly. This is mainly related to reduction in local and regional pollution sources in the past 20 years.     

Imposex in Morula Granulata as Bioindicator of Tributyltin (TBT) Contamination in the Dampier Archipelago, Western Australia

This study was undertaken to examine the use of imposex in the tropical neogastropod whelk Morula granulata as bioindicator of tributyltin (TBT) contamination. Samples were collected from sites throughout the Dampier Archipelago; both impact sites close to intensive shipping activity and remote control sites. TBT in this area originates mainly from commercial vessels using the Port of Dampier that are >25 m in length and exempt from legislation controlling the use of TBT in antifouling paints. A field survey was undertaken in July 1997; 100 M. granulata were taken at each of 18 sites with varying vessel activity, and examined for imposex. The level of TBT contamination in the Archipelago was low; ranging from <0.3 to 25 ng Sn l⁻¹ in water,<0.3 to 80 ng Sn g−¹ in oyster tissue (Saccostrea cuccullata – prey of M. granulata), and <0.3 to 33 ng Sn g⁻¹ in whelks. Percentage of imposex in M. granulata ranged from 0% to 57%, and was correlated with distance to vessel activity (r=0.646, p<0.05). The only significant relationship of imposex with butyltin concentrations was with TBT in oysters (r=0.826, p<0.05), and dibutyltin (DBT) in whelks (r=0.783, p<0.05). It is postulated that this is because of interspecies variation in ability to metabolize TBT to DBT and MBT (monobutyltin), as well as induction of imposex in earlier life stages, so that imposex in adults could be a reflection of prior contamination. M. granulata is a suitable bioindicator of TBT contamination in the Dampier Archipelago, which could be used throughout the Indo-Pacific given its abundance in the region.     

Effects of copper on survival, development and growth of Metapenaeus ensis larvae and postlarvae (decapoda: Penaeidae)

Effects of copper on survival, development and growth of the early developmental stages of the penaeid shrimp, Metapenaeus ensis, were studied in the laboratory. High variability in survival rates were observed among animals from different spawners. Larvae exposed to 0.06 and 0.10 mg Cu l⁻¹ over a 10-day period had lower survival rates and exhibited slower development than control animals. No significant treatment effects were found at 0.04 and 0.08 mg Cu l⁻¹. Effects of copper on survival and growth of postlarvae were studied over an 8-day period. Copper concentrations lower than 0.10 mg l⁻¹ did not affect survival rates. Slower growth rates were observed in postlarvae exposed to 0.08 and 0.10 mg Cu l⁻¹.     

Levels of chlorinated hydrocarbons and trace metals in bivalves and nearshore sediments from the Dominican Republic

Bivalves (Crassostrea rizhophorae, Isognomon alatus, Codakia orbicularis Linné, and Tellina fausta) and sediment samples collected from 12 locations along the Dominican Republic coastline were used to assess the degree of nearshore contamination by selected chlorinated hydrocarbons and heavy metals. Concentrations of total chlordanes, DDTs and PCBs ranged from 0.51 to 7.47 ng/g, < MDL to 30.9 ng/g and 11.3 to 82.3 ng/g, and from < MDL to 1.73 ng/g, 0.21 to 12.5 ng/g and 0.46 to 41.9 ng/g in bivalves and surface sediments, respectively. The concentration ranges of predominantly anthropogenic Cu, Pb and Zn were 3.08–866 μg/g, 0.08–1.46 μg/g and 22.9–4380 μg/g in bivalve soft parts and 1.01–111 mg/g, 0.420–134 μg/g and 1.96–277 μg/g in surface sediments, respectively. Mercury concentrations ranged from 0.290 to 7.02 μg/g and from 0.096 to 0.565 μg/g in both matrices. The relatively high Hg concentrations found in bivalves from two stations in Bahia Samana, near to a known source of past Hg contamination in the sediments, require a more in-depth survey of the area. In general the concentrations of selected chlorinated hydrocarbons and trace metals, compared with those reported for the same matrices in the Caribbean and west Atlantic coastal areas, show at present the existence of only very localized contamination problems in the Dominican Republic.     

Deuterium tracing for the estimation of transpiration from trees Part 3. Measurements of transpiration from Eucalyptus plantation, India

Measurements of transpiration from individual trees of Eucalyptus from plantations at four different sites in Karnataka, Southern India, are presented. These show large (as much as tenfold) differences in the transpiration between premonsoon and postmonsoon periods, a reflection of the effects of soil-moisture stress in the premonsoon periods. For trees with diameters at breast height (DBH) less than 10 cm the transpiration rate of individual trees is proportional to the square of the DBH. For trees which are not experiencing soil-water stress the daily transpiration rate of individual trees, q, is well represented by the relation: q = (6.6 ± 0.3)g (m³ day⁻¹ where g (m²) is the tree basal area. On a unit ground area basis the transpiration rate, expressed as a depth per day is given by the relation: E_t = (0.66 ± 0.03)G (mm day⁻¹ where g(m²ha⁻¹) is the total basal area per hectare. For all the sites studied, although there is evidence for the ‘mining’ of soil water as roots penetrate deeper depths in the soil each year, there is no evidence for direct abstraction from the water table.     

五大连池火山区地震体波Q值特征

基于五大连池火山区域地方震资料,选取信噪比高、定位准确且震中位于火山区域的39个地震,根据地震波传播的三段几何衰减模型,使用地震记录位移谱,去除仪器响应、背景噪声,反演求取各频点f的S波Q值,拟合获得火山区体波Q值与f的关系为Q（f）=221.7f^0.452,计算得到老黑山、火烧山附近区域Q值与f的关系为Q（f）=152f^0.600,与国内外其他区域体波Q值进行对比,并结合火山区地质构造活动,分析五大连池火山区体波Q值特征。     

Time should be considered in developmental ecotoxicity test

Developmental toxicity tests are often used for the hazard assessment of chemicals and environmental media. One of the most widely used is the oyster embryo larval test (OEL), in which the development of oyster larvae is arrested at a single fixed time (e.g. 24 or 48 h) of toxic exposure, and the proportion of normal larvae measured. However, a major problem with this conventional approach is the lack of information on temporal trends in development. In this study, Pacific oyster Crassostrea gigas embryos were exposed to nominal concentrations of copper (CuSO₄) of <0.001 (control), 0.60, 1.25, 2.5, 5.0, 10.0 and 20.0 μg l⁻¹ (at 20 °C, salinity 35‰ and pH 8.1). Three replicates from each group were arrested and examined every 8 h during 24–72 h of exposure, and the number of viable larvae developed to D-shape was determined. The results revealed that the number of viable D-shape larvae in the control increased rapidly and reached an optimum at 32 h, before declining gradually due to starvation. Analysis of covariance (ANCOVA) showed that larval developmental rates during 0–32 h were significantly inhibited by Cu at all concentrations. This paper demonstrates that arrest and measurement at different time periods are important and should be incorporated into the OEL test. This would maximise the sensitivity of the test in detecting developmental effects in spiked or environmental samples.