Water characterization in three industrialized harbours (Vigo, Bilbao and Pasajes) in north coast of Spain

Total dissolved Cu, Zn, Pb and Cd concentrations, speciation, fluorescence of organic matter and toxicity for sea urchin development have been studied during three years in water samples of three of the most industrialized harbours in North Coast of Spain: Vigo, Bilbao and Pasajes. Total metal concentrations were below 0.4 nM for Cd and generally below 2 nM for Pb. Zn and Cu levels were higher (2.8-9 nM Cu and 15-234 nM Zn) and denoted an anthropogenic enrichment. Ligand concentrations were usually between 50 and 200 nM for Cu and between 6 and 80 for Pb and Zn. Organic matter and speciation analysis yielded complementary information to characterize the samples, but no correlation was present between the parameters calculated by both methods. Samples were tested for toxicity with sea urchin embryo bioassay and three samples were toxic but the toxicity could not be explained with the metal levels here measured.     

A toxicity identification evaluation of silty marine harbor sediments to characterize persistent and non-persistent constituents

Sediment toxicity in silty marine harbor sediments is frequently dominated by ammonia or sulfide, leaving the adverse effects of persistent toxic substances unnoticed. To investigate the latter, we subjected interstitial water from three contaminated silty sediments to toxicity identification evaluation (TIE) phase I manipulations and tested for toxicity with four bioassays: the amphipod Corophium volutator (survival as an endpoint), the sea urchin Psammechinus miliaris (fertilization, embryo development) and the bacterium Vibrio fischeri (bioluminescence inhibition).The graduated pH manipulations identified the prominent toxicity of ammonia in the amphipod and sea urchin embryo tests, and also sulfide toxicity in the bacterium test. In two of the three samples tested with the amphipods, sea urchin embryos and bacteria, a small but significant reduction in interstitial water toxicity was achieved by removing persistent compounds through C(18) solid phase extraction. EDTA chelation resulted in a slight detoxification of the interstitial water for the amphipods and sea urchin embryos, but this was not related to any measured trace metals. Despite the presence of toxic levels of ammonia and sulfide in the harbor sediments, we established the adverse biological effects of persistent constituents by means of the TIE manipulations and in vivo interstitial water bioassays.     

Comparative performances of eggs and embryos of sea urchin (Paracentrotus lividus) in toxicity bioassays used for assessment of marine sediment quality

The potential toxicity of sediments from various ports was assessed by means of two different liquid-phase toxicity bioassays (acute and chronic) with embryos and eggs of sea urchin Paracentrotus lividus. Performances of embryos and eggs of P. lividus in these bioassays were compared for their interchangeable applicability in integrated sediment quality assessment. The obtained endpoints (percentages of normally developed plutei and fertilized eggs) were linked to physical and chemical properties of sediments and demonstrated dependence on sediment contamination. The endpoints in the two bioassays were strongly correlated and generally exhibited similar tendency throughout the samples. Therein, embryos demonstrated higher sensitivity to elutriate exposure, compared to eggs. It was concluded that these tests could be used interchangeably for testing toxicity of marine sediments. Preferential use of any of the bioassays can be determined by the discriminatory capacity of the test or vulnerability consideration of the test subject to the surrounding conditions.     

Contamination status and accumulation profiles of organotins in sea otters (Enhydra lutris) found dead along the coasts of California, Washington, Alaska (USA), and Kamchatka (Russia)

Organotin compounds (OTs) including mono- to tri-butyltins, -phenyltins, and -octyltins were determined in the liver of adult sea otters (Enhydra lutris) found dead along the coasts of California, Washington, and Alaska in the USA and Kamchatka, Russia. Total concentrations of OTs in sea otters from California ranged from 34 to 4100ng/g on a wet weight basis. The order of concentrations of OTs in sea otters was total butyltins>total octyltins> or = total phenyltins. Elevated concentrations of butyltins (BTs) were found in some otters classified under 'infectious-disease' mortality category. Concentrations of BTs in few of these otters were close to or above the threshold levels for adverse health effects. Total butyltin concentrations decreased significantly in the livers of California sea otters since the 1990s. Based on the concentrations of organotins in sea otters collected from 1992 to 2002, the half-lives of tributyltin and total butyltins in sea otters were estimated to be approximately three years.     

The effects of Esso Corexit 9527 on the fertilizing capacity of spermatozoa

The water-soluble oil dispersant Esso Corexit 9527 has earlier been found to interfere, even in low concentrations, with fertilization and development. Further studies of the effect on sea urchin spermatozoa demonstrate that Corexit 9527 gives negative biological effects in concentrations down to 0.0003 ppm.     

Toxic Effects of Cadmium on Fertilizing Capability of Spermatozoa, Dynamics of the First Cleavage and Pluteus Formation in the Sea Urchin Anthocidaris crassispina (Agassiz)

Effects of cadmium on the fertilizing capability of spermatozoa, dynamics of the first cleavage and pluteus formation in the sea urchin, Anthocidaris crassispina, were investigated. Exposure to cadmium concentrations of 0.32, 0.56, 1.0, 1.8, 3.2, 5.6 and 10.0 mg l⁻¹ for 30 min caused a significant adverse effect on the fertilizing capability of spermatozoa (EC₅₀=1.7 mg l⁻¹). For the first cleavage kinetics assay, eggs were pre-exposed to 0, 0.18, 0.32, 0.56, 1.0, 1.8, 3.2 and 5.6 mg l⁻¹ of cadmium for 30 min. Fresh sperms of fixed concentrations were then added to the eggs and the percentage of divided zygotes estimated. Concentration-dependent toxic effects on the dynamics of the first cleavage in A. crassispina were observed between 0.18 and 5.6 mg l⁻¹ cadmium. Calculation of EC₅₀ values by probit analysis at different stages of the first cleavage revealed that EC₅₀ values increased gradually from 0.37 mg l⁻¹ at the beginning to 2.24 mg l⁻¹ at the end of the cleavage process. Different larval classes (normal, abnormal and retarded plutei, preplutei, dead embryos/larvae) of A. crassispina obtained from eggs pre-treated with cadmium for 30 min and then exposed to cadmium throughout embryogenesis were also examined. The percentage of abnormal plutei was significantly higher than that of the control at 0.56 mg l⁻¹ cadmium and higher. The 48 h EC₅₀ calculated for the number of normal plutei was 1.5 mg l⁻¹. A χ²-test revealed significant changes in the distribution of proportions of different larval classes only at cadmium concentrations 1.8 mg l⁻¹. At 5.6 mg l⁻¹ of cadmium, small malformed plutei, and oval embryos with rudimentary skeleton, poorly differentiated intestines and no mouth (pre-pluteus stage) were observed. The relative sensitivity of various end points to cadmium is also compared and discussed.     

Effect of sea cucumber (Australostichopus mollis) grazing on coastal sediments impacted by mussel farm deposition

Deposit-feeding holothurians are important processors of surface sediments in many coastal marine systems. The present study examined the effect of grazing by the sea cucumber Australostichopus mollis on sediment impacted by green-lipped mussel biodeposits (faeces and pseudofaeces) from coastal aquaculture activities. Grazing effects were investigated in a series of tank-based feeding experiments conducted over 1, 2, 4 and 8 week periods. Sediment quality indicators routinely applied to determine the impacts of coastal aquaculture were used to evaluate sediment health from grazed and ungrazed sediments. Sea cucumber grazing resulted in reductions in total organic carbon, chlorophyll a and phaeopigment, as well as chlorophyll a/phaeopigment ratio of impacted sediments. These results demonstrate that sea cucumber grazing significantly reduces the accumulation of both organic carbon and phytopigments associated with biodeposition from mussel farms. Sea cucumber grazing offers a means of constraining or reversing the pollutive impacts of coastal bivalve aquaculture.     

Effects of new antifouling compounds on the development of sea urchin

Tributyltin oxide (TBTO) has been used worldwide in marine antifouling paints as a biocide for some time. However, it produced toxic effects, especially in marine water/sediment ecosystems. Consequently, its use in antifouling paints has been prohibited in many countries. In this study, the toxicity of alternative and/or new antifouling biocides compared with TBTO is assessed by a biological method. The effects of these chemicals on marine species have not been well studied. This paper assesses, comparatively, the effects of eight biocides on sea urchin eggs and embryos. The chemicals assessed were TBTO, Irgarol 1051, M1 (the persistent degradation product of Irgarol), Diuron, zinc pyrithione, 'KH101', 'Sea-Nine 211', and copper pyrithione. For these chemicals, toxicity appears to be in the order zinc pyrithione > Sea-Nine 211 > KH101 > copper pyrithione > TBTO > Diuron approximately = Irgarol 1051 > M1. Here, we show that zinc pyrithione, Sea-Nine 211, KH101, and copper pyrithione are much more toxic to sea urchins than TBTO or the other chemicals.     

Polybrominated diphenyl ethers (PBDEs) in sediments and mussel tissues from Hong Kong marine waters

Sediments and green-lipped mussels, Perna viridis, were used to investigate concentrations of polybrominated diphenyl ethers (PBDEs) in Hong Kong's marine environment. PBDEs have been used extensively over the past two decades as flame retardants in polymer additives for a variety of plastics, computers, furniture, building materials, and fabrics. Many measurements of PBDEs in various environmental matrices have been reported from Belgium, Holland, Japan, Europe and North America, but few measurements are available for the southeast Asian region and Hong Kong. PBDE congeners (n=15) were measured in 13 sediments and nine mussel samples, taken from Hong Kong marine waters. The Sigma15PBDEs in sediments ranged between 1.7 and 53.6 ng g(-1) dry wt, with the highest concentrations located around the most heavily populated areas of Victoria Harbour and Sai Kung, while the lowest concentrations of Sigma15PBDEs were found at more remote locations of Sha Tau Kok, Wong Chuk Bay, Castle Peak Bay, and Gold Coast. Sigma15PBDEs ranged from 27.0 to 83.7 ng g(-1) dry wt of mussel tissues. Although not identical, most of the congeners in sediments were found in mussel tissues, with BDE-47, BDE-99, BDE-153 and BDE-183 being the most prominent in both matrices. On the basis of a literature survey, the concentrations of PBDEs reported in Hong Kong sediments and mussel tissues are amongst the highest in the world.     

Molecular characterizations of oxytetracycline resistant bacteria and their resistance genes from mariculture waters of China

Oxytetracycline-resistant bacteria were isolated from a mariculture farm in China, and accounted for 32.23% and 5.63% of the total culturable microbes of the sea cucumber and the sea urchin rearing waters respectively. Marine vibrios, especially strains related to Vibrio splendidus or V. tasmaniensis, were the most abundant resistant isolates. For oxytetracycline resistance, tet(A), tet(B) and tet(D) genes were detected in both sea cucumber and sea urchin rearing ponds. The dominant resistance type for V. tasmaniensis-like strains was the combination of both tet(A) and tet(B) genes, while the major resistance type for V. splendidus-like strains was a single tet(D) gene. Most of the sea cucumber tet-positive isolates harbored a chloramphenicol-resistance gene, either cat IV or cat II, while only a few sea urchin tet-positive isolates harbored a cat gene, actually cat IV. The coexistence of tet and cat genes in the strains isolated from the mariculture farm studied was helpful in explaining some of the multi-resistance mechanisms.     

Assessment of the Comparative Toxicity of Sewage Effluent from 10 Sewage Treatment Plants in the Area of Sydney, Australia using an Amphipod and Two Sea Urchin Bioassays

In 1995, Sydney Water Corporation undertook an ecological and human health risk assessment for 10 sewage treatment plants (STPs) that discharge primary (6 STPs) and secondary (4 STPs) treated effluents into coastal waters in the Sydney and Illawarra regions, NSW, Australia. A program of toxicity testing of effluent from the 10 STPs was undertaken to determine the toxic effects of the effluents and as a weight-of-evidence study for the risk assessment.

Three types of bioassays were used to test multiple samples of effluent from each STP. Tests used were the sea urchin (Heliocidaris tuberculata) fertilization and larval development bioassays and the amphipod (Allorchestes compressa) survival bioassay.

Comparing between STPs, primary treated effluent was slightly more toxic than secondary treated effluent for all three tests. The highest toxicity was recorded for those STPs that use primary treatment and discharge to the shoreline. Comparing between tests, the sea urchin fertilization bioassay was slightly more sensitive than the sea urchin larval development bioassay, with the amphipod survival bioassay being the least sensitive.

This paper will describe the results of this toxicity testing program and discuss the application of the study results as weight-of-evidence for the risk assessment study.     

Toxicities of ten metals to Crassostrea gigas and Mytilus edulis embryos and Cancer magister larvae

Pacific oyster (Crassostrea gigas Thunberg) embryos, bay mussel (Mytilus edulis Linnaeus) embryos, and Dungeness crab (Cancer magister Dana) zoea I stage larvae were exposed to arsenic, cadmium, chromium, copper, lead, mercury, nickel, selenium, silver, and zinc salts. The two bivalve species were exposed through the completion of embryogenesis (48 h), and the effects of the metals upon abnormal development were monitored. The crab larval stages were exposed to the metal solutions for 96 h. Concentrations (μg l_.^?1 as metal) which caused 50% abnormal development in the two species of bivalve larvae and which were fatal to 50% of the crab larvae ranged from 5.3 (oyster) to 49 (crab) for copper; 5.8 (mussel) to 8.2 (crab) for mercury; 14 (mussel) to 55 (crab) for silver; 119 (oyster) to 456 (crab) for zinc; 232 (crab) to 3000 (mussel) for arsenic; 247 (crab) to 1200 (mussel) for cadmium; 349 (oyster) to 4360 (crab) for nickel; 476 (mussel) to 758 (oyster) for lead; 3440 (crab) to 4538 (oyster) for hexavalent chromium; and selenium greater than 10 000 for the three species. The acute toxicity testing of C. gigas embryo together with two California native species confirmed its usefulness for predicting toxicity to the latter species.     

Evaluation of the mussel Perna perna as a biomonitor of polycyclic aromatic hydrocarbon (PAH) exposure and effects

The mussel Perna perna was used to monitor and assess areas suspected of oil contamination by transplanting animals from unimpacted to impacted sites and vice versa. The deployment lasted for three months during which total PAH were measured in seawater samples collected every 15 days. PAH concentrations were determined in mussel tissue monthly. The lysosomal neutral red assay and the condition index were used as biomarkers of the adverse biological effects. The PAH concentration in mussels at unimpacted sites, prior to deployment was ca. 100 microg kg(-1) but increased to 300 microg kg(-1) after three months at the contaminated site. Mussels transplanted to the uncontaminated site showed a decrease from 380 to 80 microg kg(-1). Significant changes in lysosomal stability occurred within one month of the beginning of the transposition; in contrast condition indexes were not correlated with the observed changes in PAH concentration.     

PCB, PCDD/F and PBDE levels and profiles in crustaceans from the coastal waters of Brittany and Normandy (France)

Polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) were analysed in the muscle of various edible marine crustaceans (spider crab, edible crab, velvet swimming crab and Norway lobster) from the Brittany and Normandy coasts (France). The highest concentrations were measured in species collected from Antifer (Seine Bay). PCB and PBDE patterns in crustacean muscles were similar and independent of the geographical area with the predominance of the high chlorinated PCBs (CB153, 138, 118 and 180), and of a few PBDE congeners (BDE47, BDE99, BDE100 and BDE28). Oppositely, dioxin contamination differed with site. The major component in crustaceans from the Seine Bay was 2378-TCDF, whereas specimens from cleaner areas had higher relative concentrations of OCDD. Finally, the comparison of the spider crab contaminant profiles to those measured in mussel and sea bass highlighted two different trends: decapod crustaceans possess relatively strong capacity to metabolise PCBs and PBDEs; however these species might be used as bioindicators for dioxin pollution monitoring in the marine coastal environment.     

Effects of seawater acidification on early development of the intertidal sea urchin Paracentrotus lividus (Lamarck 1816)

The effect of pH ranging from 8.0 to 6.8 (total scale - pH_T) on fertilization, cleavage and larval development until pluteus stage was assessed in an intertidal temperate sea urchin. Gametes were obtained from adults collected in two contrasting tide pools, one showing a significant nocturnal pH decrease (lowest pH_T = 7.4) and another where pH was more stable (lowest pH_T = 7.8). The highest pH_T at which significant effects on fertilization and cleavage were recorded was 7.6. On the contrary, larval development was only affected below pH_T 7.4, a value equal or lower than that reported for several subtidal species. This suggests that sea urchins inhabiting stressful intertidal environments produce offspring that may better resist future ocean acidification. Moreover, at pH_T 7.4, the fertilization rate of gametes whose progenitors came from the tide pool with higher pH decrease was significantly higher, indicating a possible acclimatization or adaptation of gametes to pH stress.     

Responses of the green-lipped mussel Perna viridis (L.) to suspended solids

Laboratory experiments were conducted to investigate the lethal and sublethal effects of suspended solids on the survival and physiological, behavourial and morphological changes of the green-lipped mussel Perna viridis collected from Tolo Harbour, Hong Kong. Results showed that P. viridis survived in all test conditions of suspended solids from 0 to 1200 mg/l over a period of 96 h. Physiological responses of the green-lipped mussel under 14-d exposure of suspended solids from 0 to 600 mg/l, followed by 14-d recovery in natural seawater, revealed no significant changes (p>0.05) in oxygen consumption and dry gonosomatic index for treatments in different concentrations of suspended solids and exposure time. Changes in clearance rate were only found to be significant (p<0.001) with exposure time. Responses in behavourial and morphological changes of the green-lipped mussel were also studied under similar experimental treatments and exposure time. Byssus production was significantly (p<0.001) related to exposure time. Gill damage, however, was significantly greater in treatments (p<0.001). Present findings suggested that P. viridis could tolerate a high level of suspended solids in the laboratory. There were dose-dependent effects of suspended solids on morphology of gill filaments. Implications of survival and responses of the green-lipped mussel to suspended solids in the marine environment are discussed.     

Comparative petroleum hydrocarbons levels and biochemical responses in mussels from hydrothermal vents (Bathymodiolus azoricus) and coastal environments (Mytilus galloprovincialis)

Aliphatic hydrocarbons and PAHs in the whole soft tissues of Bathymodiolus azoricus from three Mid-Atlantic Ridge hydrothermal vents (Menez-Gwen, Lucky Strike and Rainbow), and Mytilus galloprovincialis from three contaminated coastal sites in South Portugal were analysed, and its effects on the digestive gland microsomes mixed-function oxygenase system (MFO) were assessed. Aliphatic hydrocarbons levels were present in the same magnitude in both coastal and hydrothermal environments, while the UCM (unresolved complex mixture) for coastal mussels were higher than in vent mussels. In general, significantly higher PAHs concentrations were found in coastal mussels, compared to B. azoricus where low molecular weight PAHs (2-3 rings) represented the majority of PAHs contrarily to what was observed in M. galloprovincialis. The MFO components were present in both mussel species, and were detected in vent mussels for the first time. However this system seems to have different roles in species from these contrasting environments. In coastal mussels MFO responded to hydrocarbon contamination while response in hydrothermal organisms appeared to be related mainly to endogenous factors.     

The role of organochlorines in cancer-associated mortality in California sea lions (Zalophus californianus)

Wild California sea lions (Zalophus californianus) have an unusually high prevalence of neoplasms (18% of stranded dead adults) and high levels of contaminants. The contribution of organochlorine (OC) tissue burdens to the probability of sea lions dying from carcinoma was explored using a logistic regression model. Levels of PCBs and DDTs were determined in blubber of sea lions diagnosed with metastatic carcinoma and animals that had died from non-carcinoma-related incidents (e.g., gunshot, domoic acid poisoning). Animals with carcinoma had higher mean concentrations (based on wet weight) of PCBs and DDTs (more than 85% and 30% higher, respectively) in blubber than did sea lions without carcinoma; the highest concentrations of OCs in the sea lions affected with carcinoma were measured in the males. Blubber thickness was significantly different between the two groups of sea lions, but after controlling for this difference, there was still a significant effect of PCBs, but not DDTs, on the probability of sea lions dying with carcinoma. Age, sex, mass and length did not affect the probability of dying from carcinoma.     

Comparison of metal accumulation between 'Artificial Mussel' and natural mussels (Mytilus galloprovincialis) in marine environments

The passive sampler called 'Artificial Mussel' (AM) developed by Wu et al. (2007) can provide a time-integrated estimate of metal concentrations in the marine environment, and offers a potential device to assess and compare metal concentration in different marine environments worldwide. The aim of this study was to compare metal accumulation on AM and natural mussel Mytilus galloprovincialis at three sites with different metal loads along the Portuguese coast for four months. M. galloprovincialis were placed in cages alongside AMs at each site. Samples were collected monthly and Cd, Cr, Cu, Pb, and Zn concentrations in whole soft tissues and AMs compared. For both Cu and Cd, the results were similar between AMs and natural mussels. Higher concentrations of Zn were observed in natural mussels, whereas the inverse was shown for Pb (about 10-fold higher). Our results showed that AMs are promising tools for assessing metal concentrations in marine environments.