The DNA de-methylating agent 5-azacytidine does not restore CYP1A induction in PCB resistant Newark Bay killifish (Fundulus heteroclitus)

Newark Bay (NB) killifish (Fundulus heteroclitus) have been chronically exposed to environmental contaminants that activate the aryl hydrocarbon receptor (AHR) and are tolerant to toxic effects and CYP1A induction provoked by AHR ligands. Resistance to CYP1A induction could be due to an epigenetic mechanism such as DNA methylation. We measured in-ovo CYP1A catalytic activity (ethoxyresorufin-O-deethylase, EROD) in NB and reference site killifish embryos aqueously exposed to various concentrations of the de-methylating agent 5-azacytidine, 5-AC (5, 50 and 500 μ(micro)M) with or without 0.2 μ(micro)g/l of the CYP1A inducer 3,3^′,4,4^′,5 pentachlorobiphenyl (IUPAC PCB126). Neither PCB126 alone, nor PCB126 plus 5-AC, induced EROD above levels in vehicle treated Newark Bay fish. In reference site fish, the same PCB126 dose provoked a 7.4-fold EROD induction relative to controls. We conclude that Newark Bay killifish are resistant to CYP1A induction by co-planar PCBs during early embryological development and our data suggests that DNA methylation does not play a critical role in resistance to CYP1A induction in this model.     

PCB uptake and accumulation by oysters (Crassostrea virginica) exposed via a contaminated algal diet

Reproductively active oysters were fed daily with 0.2 g algal paste containing 0, 0.1, and 1.0 μg polychlorinated biphenyls (PCBs) (1:1:1 mixture of Aroclor 1242, 1254 and 1260) for either 15 or 30 days, and accumulation of PCBs in different organ tissues and eggs assessed. The effects of PCB exposure on lipid content, lipid class and fatty acid composition were also evaluated. PCBs were accumulated by the oysters and transferred to the eggs. PCB accumulation in oysters was dose, time and tissue dependent. Mean PCB contents were 3150, 1970, and 250 ng/g dry wt., respectively, in the visceral mass, gills+mantle and muscle of oysters fed algal paste containing 1.0 μg PCBs for 30 days. The PCBs in the eggs from the same oysters reached 671 ng PCBs/g dry wt. Feeding oysters with PCB-sorbed algal paste for 30 days significantly increased phospholipid and free fatty acid contents in gills+mantle tissues compared to the same tissues in the undosed control.     

Transport and accumulation of organochlorines in the ovaries of Atlantic croaker (Micropogonias undulatus)

The role of vitellogenin in the transport of organochlorines was investigated in Atlantic croaker (Micropogonias undulatus) by exposing them through the diet to o,p′-DDT at a concentration of 10.8 μg/100 g fish/day for 14 days or Aroclor 1254 (PCB) at a concentration of 0.5 mg/100 g fish/day for 30 days during gonadal recrudescence. Tissue samples were taken from the fish at various times after initial exposure, and o,p′-DDT and PCBs were extracted in acetonitrile and analyzed by gas chromatography. Analysis of the ovarian tissue collected 2 and 3 weeks from the start of exposure revealed that the o,p′-DDT concentration increases as the gonadosomatic index (GSI) increases (r² = 0.63), with accumulation ranging from less than 1% to as much as 8% of the total dosage. Interestingly, o,p′-DDT did not accumulate in the testes during the same exposure period. Accumulation of PCBs was found to be 40 times higher in the ovaries than in the testes. Gel filtration of plasma from exposed females showed that o,p′-DDT elutes in the low density lipoprotein and vitellogenin fractions. Control plasma incubated with o,p′-DDT at 4 °C for 16 h followed by chromatography on Sepharose 6B gave similar results with an o,p′-DDT concentration of 0.6μg/mg protein in the vitellogenin fraction. Furthermore, both o,p′-DDT and PCBs were found to bind to purified croaker vitellogenin. These results suggest that lipoproteins, including vitellogenin, are involved in the transport and accumulation of organochlorines in the ovaries of exposed fish.     

Recent trends in organochlorine residues in mussels (Mytilus edulis) from the Mersey Estuary.

Blue mussels (Mytilus edulis) taken from several sites in the Mersey Estuary, an urban-industrial water body in NW England, have been analyzed for residues of the persistent organochlorines, DDT, PCB congeners nos. 28, 52, 101, 118, 138, 153 and 180 (ICES7), and alpha-MHCH (alpha-methyl hexachlorocyclohexane). The concentration range for sigmaPCB (ICES7) was 13.9-34.9 microg kg(-1) in 1994 compared with 9.6-31.9 microg kg(-1) in 1998, whilst the equivalent concentrations of sigmaDDT were 8.9-32.4 microg kg(-1) and 4.5-16.8 microg kg(-1), respectively. The concentration of alpha-MHCH, an organochlorine largely restricted to the Mersey Estuary, was 1.2-11 microg kg(-1) in 1994 and 0.3-1.3 microg kg(-1) in 1998. At both sampling dates the inner estuary sites of Rock Ferry and Egremont were more contaminated than the outer estuary sites of New Brighton. Dove Point and Caldy Blacks. The lower concentrations of organochlorines in mussels in 1998 compared with 1994 are thought to be early evidence of significant improvements in water quality derived from major capital investment in the treatment of industrial effluents and sewage wastewater, linked to a combination of stricter legislative controls and the environmental benefits from the adoption of new, clean technologies in manufacturing.     

Distribution, condition, and growth of newly settled southern flounder (Paralichthys lethostigma) in the Galveston Bay Estuary, TX

Several flatfish species, including southern flounder (Paralichthys lethostigma) recruit to estuaries during early life. Therefore, evaluation of estuarine sites and habitats that serve as nurseries is critical to conservation and management. The present study used density data in conjunction with biochemical condition and growth measurements to evaluate settlement sites used by southern flounder in the Galveston Bay Estuary (GBE). In 2005, beam-trawl collections were made in three major sections of the GBE (East Bay, Galveston Bay, West Bay). Three sites were sampled in each bay. Within each sampling site, replicate collections were taken from three habitats: 1) marsh edge (< 1 m depth), 2) intermediate zone (10–20 m from marsh interface;  1 m depth), and 3) bay zone (typically > 100 m from marsh interface; depth > 1 m). Average size of southern flounder collected was 12–19 mm standard length, and peak densities occurred in January and February. Catch data indicated that densities of southern flounder were significantly greater in East Bay (2.75 per 100 m²) than in Galveston Bay (0.91 per 100 m²) or in West Bay (0.45 per 100 m²). Densities were statistically similar among habitats. Otolith-based estimates of age indicated that the majority of southern flounder collected were 35–45 days old and derived from early December to early January hatch-dates. Growth rates were similar among bays and among habitats, with the average growth rate being 0.40 mm day^− 1 (range: 0.21–0.76 mm day^− 1). RNA:DNA was above the established baseline value for nutritional stress, indicating that newly settled southern flounder in the GBE were in relatively high condition. Habitat-specific differences in RNA:DNA ratios were not observed; however, ratios were significantly lower in West Bay (average 8.0) than in East Bay (average 9.5) or in Galveston Bay (average 9.8), suggesting the condition of new recruits may vary spatially within the GBE. Findings from the current study suggest density and condition of newly settled southern flounder vary at the bay scale, suggesting that parts of GBE do not function equally as nurseries.     

Municipal wastewater contamination in the southern california bight: Part I—metal and organic contaminants in sediments and organisms

Sediments and organisms were examined for concentrations of organic and metal contaminants from near the Los Angeles County (JWPCP) municipal outfall at Palos Verdes (PV) station 7-3, the Los Angeles City (Hyperion) municipal outfall at Santa Monica Bay (SMB) station 6-4 and reference station SMB 2–3 near Malibu Beach. Flows and mass emission rates of suspended solids, PCBs, Cd and Zn were similar at the two outfalls. Mass emission rate of copper was almost twice as high from Hyperion as from JWPCP, while mass emission rate of DDTs was an order or magnitude higher from JWPCP than from Hyperion.Surficial sediments at PV 7-3 were enriched in most contaminants relative to SMB 6-4 and relative to the mass emission rates of contaminants from the JGVPCP and Hyperion outfalls. Some of this enrichment could be accounted for by the greater accumulation of organic material, measured as total volatile solids, at PV 7-3 relative to SMB 6-4. Some might be accounted for by resurfacing of more contaminated historical deposits buried at PV 7-3. Some of the enrichment of DDTs relative to PCBs could be accounted for by the greater abundance of oxygenated metabolites of PCBs (PCBols) relative to DDTs (DDTols) in sediments.The degree of contamination of organisms by DDTs increased with proximity to PV 7-3 but contamination by PCBs was similar at PV 7-3 and SMB 6-4. DDT concentrations in fish livers ranged from 12 ± 4 ( ) mg/wet kg in longspine combfish from SMB 2–3 to 610 ± 105 (n = 5) mg/wet kg in Pacific sanddab from PV 7-3. DDT concentrations in fish gonads ranged from 0·003 ± 0·003 (n = 5) mg/wet kg in yellowchin sculpin from SMB 6-4 to 1.5 ± 6 (n = 3) mg/wet kg in Pacific sanddabs from PV 7-3. PCB concentrations in fish livers ranged from 1·2 ± 0·4 (n = 4) mg/wet kg in yellowchin sculpin from SMB 2–3 to 16 ± 3 (n = 6) in Pacific Sanddab from SMB 6-4. DDT and PCB concentrations in invertebrate hepatopancreas were only slightly lower than those in fish livers. DDTols and PCBoIs comprised an average of 91 % of the total of parent compounds and oxygenated metabolites in sediments and 66 % in livers and hepatopancreas. Trace metals were frequently decreased in livers and hepatopancreas from near outfalls even though they were highly elevated in sediments.Comparison of sediment and tissue chlorinated hydrocarbon data with that from Elliot and Commencement Bays, Puget Sound, indicated that none of the southern California coastal stations considered in this study were sufficiently lacking contamination to be considered as adequate control sites.     

Factors affecting the elimination of PCBs in the marine copepod Acartia tonsa

The effects of feeding, egg laying, and fecal pellet production on the elimination of polychlorinated biphenyls (PCBs) from the marine copepod Acartia tonsa were studied in a series of experiments. Copepods were exposed to ¹⁴C-labelled Aroclor 1254 and allowed to depurate in clean seawater. Copepods fed during depuration eliminated PCBs more rapidly than unfed copepods whether or not the original PCB exposure medium had contained food. Both eggs and fecal pellets contained PCBs during depuration, with the weight specific concentration of PCB in the eggs (up to 407 ppm, dry weight) exceeding four times that in the females that produced them. Female copepods eliminated PCBs twice as rapidly as males, indicating that egg production is an important route for PCB elimination.     

Enrichment of organochlorine contaminants in the sea surface microlayer: An organic carbon-driven process

Over 50 seawater samples from two different sites—Barcelona (Spain) and Banyuls-sur-Mer (France)—were analyzed in order to study the extent and postulate the processes driving the enrichment of hydrophobic organic pollutants in the sea surface microlayer (SML). A number of individual polychlorinated biphenyl (PCB) congeners (41) were measured to study their partitioning between the particulate (fraction > 0.7 μm) and the dissolved + colloidal phases (fraction < 0.7 μm), with the latter being differentiated into estimated dissolved and colloidal phases. In addition, several organochlorine pesticides were also measured, namely, HCB, α-HCH, γ-HCH, 4,4′-DDE, 4,4′-DDD and 4,4′-DDT. The presence of PCB congener profiles found in the SML suggests a dynamic coupling with the atmosphere in Banyuls sampling site, whereas offshore Barcelona the presence of highly chlorinated congeners was due to persistent sediment resuspension. The average PCB concentration in the SML dissolved + colloidal phase were higher in Banyuls (7.8 ng L^{− 1}) than in Barcelona (3.6 ng L^{− 1}) samples, but in the particulate phase concentrations were higher in Barcelona (3.2 ng L^{− 1}) to that of Banyuls (1.4 ng L^{− 1}). However, PCB concentrations in the SML generally also showed large variability. Enrichment factors of PCBs and other organochlorine compounds in the SML with respect to the underlying water column ranged from 0.2 to 7.4. This may be explained for both the dissolved + colloidal and particulate phases by the enrichment in the SML of organic carbon (OC) as discerned from particle–water and colloid–water partitioning.     

Variability of organic δC and C/N in the Mersey Estuary, U.K. and its implications for sea-level reconstruction studies

Microfossil analysis (e.g. diatoms, foraminifera and pollen) represents the cornerstone of Holocene relative sea-level (RSL) reconstruction because their distribution in the contemporary inter-tidal zone is principally controlled by ground elevation within the tidal frame. A combination of poor microfossil preservation and a limited range in the sediment record may severely restrict the accuracy of resulting RSL reconstructions. Organic δ¹³C and C/N analysis of inter-tidal sediments have shown some potential as coastal palaeoenvironmental proxies. Here we assess their viability for reconstructing RSL change by examining patterns of organic δ¹³C and C/N values in a modern estuarine environment. δ¹³C and C/N analysis of bulk organic inter-tidal sediments and vegetation, as well as suspended and bedload organic sediments of the Mersey Estuary, U.K., demonstrate that the two main sources of organic carbon to surface saltmarsh sediments (terrestrial vegetation and tidal-derived particulate organic matter) have distinctive δ¹³C and C/N signatures. The resulting relationship between ground elevation within the tidal frame and surface sediment δ¹³C and C/N is unaffected by decompositional changes. The potential of this technique for RSL reconstruction is demonstrated by the analysis of part of an early Holocene sediment core from the Mersey Estuary. Organic δ¹³C and C/N analysis is less time consuming than microfossil analysis and is likely to provide continuous records of RSL change.     

Sources and biogeochemical cycling of particulate selenium in the San Francisco Bay estuary

As part of a study of estuarine selenium cycling, we measured the concentration, chemical form (speciation), and distribution of particulate selenium under various river flow conditions in the North San Francisco Bay (from the Golden Gate to the Sacramento and San Joaquin Rivers). We also conducted laboratory studies on the accumulation of selenium by phytoplankton, the critical first step in the transformation of dissolved to particulate selenium. Total particulate selenium concentration in the North SF Bay was relatively constant between high and low flow periods, ranging spatially from 0.05 to 0.35 nmol l⁻¹ and comprising between 5 and 12% of the total water column selenium inventory. Mean concentrations were generally highest in the Carquinez Strait–Suisun Bay region (salinity 0–17) and lowest in Central Bay. However, selenium content of suspended particles varied with river flow, with higher content during low flow (9.76 ± 4.17 nmol g⁻¹; mean ± sd; n = 67) compared to high flow (7.10 ± 4.24 nmol g⁻¹; n = 39). Speciation analyses showed that most particulate selenium is organic selenide (45 ± 27%), with a smaller proportion (typically <30%) of adsorbed selenite + selenate and a varying proportion (35 ± 28%) of elemental selenium. Based on the amount of elemental selenium in the seston (total suspended material), we calculate that resuspension of estuarine sediments could contribute 29–100% of particulate selenium in the water column. While selenium content of SF Bay seston (>0.4 μm) is relatively unenriched compared to phytoplankton (13.6–155 nmol g⁻¹ dry weight) on a mass basis, when normalized to carbon or nitrogen, seston contains a similar selenium concentration to SF Bay sediments or phytoplankton cultures. SF Bay seston is thus comprised of selenium-rich phytoplankton and phyto-detritus, but also inorganic clay mineral particles that effectively “dilute” total particulate selenium. Selenium concentrations in algal cultures (11 species) exposed to 90 nmol l⁻¹ selenite show relatively large differences in selenium accumulation, with the diatoms, chlorophytes and cryptophytes generally having lower selenium cell content (3.8 ± 2.7 × 10⁻⁹ nmol selenium cell⁻¹) compared to the dinoflagellates (193 ± 73 × 10⁻⁹ nmol selenium cell⁻¹). Because phytoplankton are such a rich (but variable) source of selenium, their dynamics could have a profound effect on the particulate selenium inventory in the North SF Bay.     

Radium mass-balance in Jamaica Bay, NY: Evidence for a substantial flux of submarine groundwater

A mass balance for the naturally-occurring radium isotopes (²²⁴Ra, ²²³Ra, ²²⁸Ra, and ²²⁶Ra) in Jamaica Bay, NY, was conducted by directly estimating the individual Ra contributions of wastewater discharge, diffusion from fine-grained subtidal sediments, water percolation through marshes, desorption from resuspended particles, and water exchange at the inlet. The mass balance revealed a major unknown source term accounting for 19–71% of the total Ra input, which could only be resolved by invoking a source from submarine groundwater. Shallow (< 2 m depth) groundwater from permeable sediments in Jamaica Bay was brackish and enriched in Ra relative to surface bay waters by over two orders of magnitude. To balance Ra fluxes, a submarine groundwater input of 0.8 × 10⁹–9.0 × 10⁹ L d^− 1 was required. This flux was similar for all four isotopes, with individual estimates varying by less than a factor of 2. Our calculated groundwater flux was 6- to 70-fold higher than the fresh groundwater discharge to the bay estimated by hydrological methods, but closely matched direct flow rates measured with seepage meters. This suggests that a substantial portion of the discharge consisted of recirculated seawater. The magnitude of submarine groundwater discharge varied seasonally, in the order: summer > autumn > spring. Chemical analyses suggest that the recirculated seawater component of submarine groundwater delivers as much dissolved nitrogen to the bay as the fresh groundwater flux.     

Distributions, sources and atmospheric fluxes of nitrous oxide in Jiaozhou Bay

The distributions, sources and atmospheric fluxes of nitrous oxide (N₂O) in the seawater of Jiaozhou Bay were investigated during four surveys in 2003 to evaluate this area as a source of N₂O to the atmosphere. N₂O concentrations in both the surface and bottom waters of Jiaozhou Bay showed obvious variability with both seasons and tidal cycles. Atmospheric fluxes of N₂O in Jiaozhou Bay showed seasonal and spatial variations, with the highest values occurring in summer and the lowest in winter. The annual emission of N₂O from the bay was estimated to be 1.09 × 10⁶–2.23 × 10⁶ mol yr⁻¹. N₂O in the water column of Jiaozhou Bay was found to come from several external sources including riverine water, sewage water and groundwater input, among which the riverine input was dominant while the groundwater input was rather limited. The spatial variation in distribution and atmospheric fluxes of N₂O in Jiaozhou Bay was influenced by the input of polluted river waters and sewage effluent along the eastern coast, which highlights the effects of human impacts on N₂O emission rates.     

The effect of PCBs on glycogen reserves in the eastern oyster Crassostrea virginica

Recent declines in Chesapeake Bay oyster populations have been attributed to disease, and reduced water quality from pollution. The stress associated with pollutant exposure may reduce energy available for growth and reproduction. Polychlorinated biphenyls (PCBs) are lipophilic contaminants that may potentially affect mobilization of lipid reserves, increasing reliance on glycogen stores, which could otherwise be utilized to supply energy for gametogenesis. Thus, PCBs may indirectly affect glycogen stores in oysters in a deleterious manner. To test for this effect, reproductively inactive oysters were exposed to PCBs by feeding individuals 0.7 g of algal paste containing 0, 0.35, or 3.5 μg PCBs daily for 8 weeks. Additionally, a group of oysters was exposed to PCBs (0, 0.35, and 3.5 μg) plus 0.3 g of non-toxic artificial sediment to examine interactive effects of sediment particles and PCBs. Adductor muscle, mantle, and gonadal tissues were analyzed for glycogen content. Results suggest that glycogen content is reduced in the adductor muscle with increasing PCB exposure, but there are no effects of PCBs in the mantle and gonadal tissues.     

Determination of adsorbed and absorbed polychlorinated biphenyls (PCBs) in seawater microorganisms

To distinguish between adsorbed and absorbed PCBs in seawater microorganisms, 2,2′,4,4′,5,5′-hexachlorobiphenyl (IUPAC #153; HCB) was added to a pure culture of Chrysochromulina apheles Moestrup et H.A. Thomsen. After the addition of the HCB, the cells were immediately harvested onto 2 μm polycarbonate filters and rinsed with a gradient of ethanol concentrations. Rinsing with 40% ethanol (v/v) was found to remove 80% of the HCB, which was loosely adsorbed to the cell surfaces, but did not extract the interior of the cells, as tested by chlorophyll a analysis. This method was used in a time course experiment which estimated PCBs adsorption and absorption to different groups of plankton organisms. Three different ¹⁴C-PCBs, 4-chlorobiphenyl (IUPAC #3; MCB), 2,2′,5,5′-tetrachlorobiphenyl (IUPAC #52; TCB), and 2,2′,4,4′,5,5′-hexachlorobiphenyl (IUPAC #153, HCB), were incubated in seawater from the northern Baltic Sea during a spring bloom. Samples were taken every third day and separated by filtration into three fractions; 0.2–2 μm (bacteria), 2–10 μm (flagellates), and >10 μm (microplankton; phytoplankton and protozoa). Two subsamples were retained from each size fraction. One of the subsamples was left untreated, to obtain adsorbed plus absorbed PCB, while the other subsample was rinsed with 40% of ethanol, to obtain the absorbed PCB. The sorption was found to vary depending on the hydrophobicity of the compounds, the structure of the cell membranes, and the lipid content and composition of the cells. The absorption increased for the TCB and the HCB in the largest size fraction over time, which coincided with an increase of the neutral and non-polar lipids.     

An analysis of the trophic network of a macrotidal estuary: the Seine Estuary (Eastern Channel, Normandy, France)

A steady state, mass balance, trophic network has been constructed to illustrate the flow of energy in the Seine Estuary by using Network Analysis and Ecopath methods. This ecosystem shows 15 compartments from primary producers to the top consumers (fish and birds). This study has been compared with other ecosystems of comparable nature located in North America (Narragansett, Chesapeake, Delaware Bays), Europe (Ems Estuary, Dublin Bay and Bay of Somme), and South Africa (Swartkops Estuary) in which analysis of trophic network has been applied with similar methods.The Seine Estuary shows values of some global parameters and indices either close to large North American bays and a South African estuary characterised by the complexity of their trophic network, or values near European bays and estuaries, or else remain typical of the Seine estuary. All of this reflects specific functioning of the Seine Estuary which can be explained by the analysis of the dominant food web. In the upstream sector an important pelagic food web was found correlated with highest primary production, especially planktonic, which is rapidly consumed by an abundant zooplankton and suprabenthos (Mysidacae and Decapoda Crustacea). This reveals the dominant consumer role of this sector. The external fluvial inputs (277.80 gC m⁻² y⁻¹) are transferred to the downstream sector which produces the major export (548.43 gC m⁻² y⁻¹), in parallel with the low consumption and efficiency of dominant benthos component from its bentho-pelagic food web. This reflects the dominant exporter role of this sector.In the Seine Estuary low values of detritivory index D/H (2.52), recycling index FCI (16.1%) and connectance (0.24) were found associated with high values of P/B ratio (38.2%), sum of exports (548.43 gC m⁻² y⁻¹) and the great difference between ratio of ascendency to capacity development A/C and internal ratio A_i/C_i. This shows the lack of a dominant resource as in Delaware Bay, that the state of development is different from a mature ecosystem, and the dependance on external connections similar to the Bay of Somme, another ecosystem of Eastern Channel, France.     

Particle–water interactions of 2,2′,5,5′-tetrachlorobiphenyl under simulated estuarine conditions

A batch sorption technique for the determination of particle–water interactions of hydrophobic organic micropollutants under simulated estuarine conditions is described. Results are presented for the behaviour of 2,2′,5,5′-tetrachlorobiphenyl (2,2′,5,5′-TCB) in river and sea waters, both in the presence and absence of estuarine suspended particles. Adsorption onto particles in sea water was enhanced compared with adsorption in river water owing to salting out of the compound, and possibly of the particulate organic matter, in the presence of high concentrations of dissolved ions. The particle–water distribution coefficient, K_D, decreased from about 120×10³ to 10×10³ ml g⁻¹, and from about 150×10³ to 20×10³ ml g⁻¹, in river water and sea water, respectively, over a particle concentration range of 10–1000 mg l⁻¹. Incomplete recovery of compound from the reactor walls is partly responsible for a particle concentration effect, while artefacts relating to inadequate sediment and water phase separation were ruled out following further experiments. The particle concentration effect, which is replicated in many field studies of hydrophobic organic micropollutants, including 2,2′,5,5′-TCB, is incorporated into a simple partitioning model and is discussed in the context of the likely estuarine behaviour of such compounds.     

Solvent extraction of copper acetylacetonate in studies of copper(II) speciation in seawater

A liquid-liquid partition, ligand exchange procedure involving the formation of copper(II) complexes with acetylacetone is presented for the determination of stability constants and concentrations of copper chelators in seawater. Acetylacetone competes with natural ligands for copper, and the equilibrium concentration of the copper acetylacetonate complex is used in speciation calculations. The concentration of the complex is calculated by partitioning a fraction of it into an organic phase and determining the total Cu concentration in that phase by back extracting with acid, and analyzing by flameless atomic absorption spectroscopy. The concentration of Cu acetylacetonate in seawater in equilibrium with the organic phase is calculated from the partition coefficient. The simple, thermodynamically well characterized procedure offers several advantages over previous techniques. Studies using organic free seawater and model ligands show good agreement between experimental and calculated conditional stability constants. Studies from seawater in Biscayne Bay, Florida, indicate two ligand types are present; type 1, K₁ = 1.2 × 10¹², C_L1 = 5.1 × 10⁻⁹ M; type 2, K₂ = 2.8 × 10¹⁰, C_L2 = 1.1 × 10⁻⁷ M. Speciation is dominated by ligand type 1. Depth profiles of [Cu(II)]_free/[Cu(II)]_total measured with the procedure at ambient copper concentrations show an increase from < 5 × 10⁻⁵ at 50–60 m to > 1 × 10⁻³ at the surface at two stations off the Florida coast.     

Total organic carbon distribution and budget through the Strait of Gibraltar in April 1998

In order to investigate total organic carbon (TOC) exchange through the Strait of Gibraltar, samples were taken along two sections from the western (Gulf of Cádiz) and eastern (Western Alboran Sea) entrances of the Strait and at the middle of the Strait in April 1998. TOC was measured by using a high-temperature catalytic oxidation method. The results referenced here are based on a three-layer model of water mass exchange through the Strait, which includes the Atlantic inflow, Mediterranean outflow and an interface layer in between. All layers were characterised by a decrease of TOC concentrations from the Gulf of Cádiz to the Western Alboran Sea: from 60–79 to 59–66 μM C in the Atlantic inflow and from 40–60 to 38–52 μM C in the Mediterranean waters, respectively. TOC concentrations in the modified North Atlantic Central Water varied from 43 to 55 μM C. Intermediate TOC values were measured in the interface layer (43–60 μM C). TOC concentrations increased from the middle of the Strait towards continents indicating a contribution of organic carbon of photosynthetic origin along Spain and Morocco coasts or TOC accumulation due to upwelling in the northeastern part of the Strait. Our results indicate that the short-term variability caused by the tide greatly impacts the TOC distribution, particularly in the Gulf of Cádiz. The TOC input from the Atlantic Ocean to the Mediterranean Sea through the Strait of Gibraltar varies from 0.9×10⁴ to 1.0×10⁴ mol C s⁻¹ (or 0.28×10¹² to 0.35×10¹² mol C year⁻¹, respectively). This estimate suggests that the TOC inflow and outflow through the Strait of Gibraltar are two and three orders of magnitude higher than reported via the Turkish Straits and Mediterranean River inputs.     

Polychlorinated biphenyls in fish-eating sea birds—molecular features and metabolic interpretations

Polychlorinated biphenyls (PCBs) are an important group of environmental pollutants, there being a total of 209 theoretical congeners. Residue analysis of the adipose tissue of five species of fish-eating sea birds from British and Irish coastal waters revealed the presence of up to 60 different congeners. By GC-MS, GC-MSD and high resolution capillary GC-ECD using authentic standards, it was possible to identify and quantify 40 different congeners. Despite the large number of PCB congeners identified only 10 accounted for > 80% of the total PCBs. A PCB congener was identified accounting for 5% of the total PCBs (2,3′,4,4′,5′,6-hexachlorobiphenyl) [168] not usually reported in biological samples. Comparison of the molecular structure for the persistent PCB congeners revealed the lack of meta-para unsubstituted adjacent carbon atoms. It has been shown that meta-para unsubstituted adjacent carbon atoms facilitate the metabolism of PCBs and it is hypothesised that the formation of hydroxy derivatives may depend upon such a requirement.     

Estuarine refugia and fish responses to a large anoxic,hydrogen sulphide, “black tide” event in the adjacent marine environment

The fish fauna of the Berg River Estuary was sampled from the mouth to 40 km upstream using a small-meshed seine-net before (summer 1993), during (summer 1994) and after (summer 1996) a low-oxygen, hydrogen sulphide “black tide” event that caused a mass mortality of fish in St Helena Bay. These data were compared to determine how the species composition, abundance and distribution of the fish fauna of the Berg River Estuary differed before, during and after the event as well as to ascertain which species, if any, found refuge in the estuary. The overall catch-per-unit-effort of 1637 fish.haul⁻¹ during the event was almost double the 932 fish.haul⁻¹ and 643 fish.haul⁻¹ in the years before and after respectively. All the fish recorded alive in the estuary during the event were species known to have some degree of estuarine association. No representatives of the purely marine species found dead on the adjacent shoreline were recorded live in the estuary during the event. Of the 10 estuarine-associated species sampled, 5 extended their range and/or modal peaks of abundance further upstream during the event. One species, Liza richardsonii, was abundant enough to examine its size distribution in different breaches of the estuary. Large/adult fish were concentrated further upstream than small/juvenile fish, which appeared to be unable to escape tidal currents and were concentrated at the edge of the low-oxygen front. Collectively this circumstantial evidence indicates that (1) fish used the Berg Estuary as a refuge from low-oxygen conditions in the marine environment during the “black tide” event, and (2) the ability to secure refuge in the estuary was restricted to species described as “estuarine-associated” or “estuarine-dependent”.