In vitro alternatives to whole animal testing. Comparative cytotoxicity studies of divalent metals in established cell lines derived from tropical and temperate water fish species in a neutral red assay

A simple vital dye (neutral red) uptake method for investigation of the acute toxicity of seven divalent metal cations to cell lines derived from the turbot (TF) and bluegill (BF-2) was evaluated. The method was extremely reproducible between replicate experiments on a week to week basis as well as being reproducible between different laboratories. Whilst there were some species-dependent differences in the relative toxicities of the metals there were good correlations between toxicity and the chemical softness (Σp) of the metal ions and also between in vivo and in vitro toxicities. These studies demonstrate that this simple and relatively inexpensive technique carried out in 96-well microplates may be extremely useful for evaluation of structure-toxicity relationships between related groups of compounds.     

Accumulation and effects of organotin compounds in oysters and mussels: Correlation with serum biochemical and cytological factors and tissue burdens

Oysters and mussels exposed to a concentration of 0·7 ppb (μg/liter) tributyltin from painted panels in flowing seawater accumulated tin in the digestive glands to comparable levels. The mussels experienced approximately 50% mortality during the 60-day test period, but the oysters suffered virtually no deaths. There was no evidence from either bivalve of elevated numbers of hemocytes during the test period and no evidence for cellular disruption as detected by increased levels of serum lysosomal hydrolases. Serum protein of exposed mussels relative to controls increased with time of exposure to the toxicant, while oyster serum protein, normally 10 x higher than in mussels, did not. No evidence was found for elevated stress proteins (heat shock proteins) or metallothioneins in the serum hemocytes of either bivalve. Responses by these animals to fatal or near fatal doses of TBT were thus very different from responses to copper that we have reported elsewhere.^1,2     

Holocene sedimentation of a wave-dominated barrierisland shoreline: Cape Lookout, North Carolina

The sedimentary record of 130 km of microtidal (0.9 m tidal range) high wave energy (1.5 m average wave height) barrier island shoreline of the Cape Lookout cuspate foreland has been evaluated through examination of 3136 m of subsurface samples from closely spaced drill holes. Holocene sedimentation and coastal evolution has been a function of five major depositional processes: (1) eustatic sea-level rise and barrier-shoreline transgression; (2) lateral tidal inlet migration and reworking of barrier island deposits; (3) shoreface sedimentation and local barrier progradation; (4) storm washover deposition with infilling of shallow lagoons; and (5) flood-tidal delta sedimentation in back-barrier environments.

Twenty-five radiocarbon dates of subsurface peat and shell material from the Cape Lookout area are the basis for a late Holocene sea-level curve. From 9000 to 4000 B.P. eustatic sea level rose rapidly, resulting in landward migration of both barrier limbs of the cuspate foreland. A decline in the rate of sea-level rise since 4000 B.P. resulted in relative shoreline stabilization and deposition of contrasting coastal sedimentary sequences. The higher energy, storm-dominated northeast barrier limb (Core and Portsmouth Banks) has migrated landward producing a transgressive sequence of coarse-grained, horizontally bedded washover sands overlying burrowed to laminated back-barrier and lagoonal silty sands. Locally, ephemeral tidal inlets have reworked the transgressive barrier sequence depositing fining-upward spit platform and channel-fill sequences of cross-bedded, pebble gravel to fine sand and shell. Shoreface sedimentation along a portion of the lower energy, northwest barrier limb (Bogue Banks) has resulted in shoreline progradation and deposition of a coarsening-up sequence of burrowed to cross-bedded and laminated, fine-grained shoreface and foreshore sands. In contrast, the adjacent barrier island (Shackleford Banks) consists almost totally of inlet-fill sediments deposited by lateral tidal inlet migration. Holocene sediments in the shallow lagoons behind the barriers are 5–8 m thick fining-up sequences of interbedded burrowed, rooted and laminated flood-tidal delta, salt marsh, and washover sands, silts and clays.

While barrier island sequences are generally 10 m in thickness, inlet-fill sequences may be as much as 25 m thick and comprise an average of 35% of the Holocene sedimentary deposits. Tidal inlet-fill, back-barrier (including flood-tidal delta) and shoreface deposits are the most highly preservable facies in the wave-dominated barrier-shoreline setting. In the Cape Lookout cuspate foreland, these three facies account for over 80% of the sedimentary deposits preserved beneath the barriers. Foreshore, spit platform and overwash facies account for the remaining 20%.     

Evidence from heterologous expression of glutathione S-transferases A and A1 of the plaice (Pleuronectes platessa) that their endogenous role is in detoxification of lipid peroxidation products

cDNA clones for glutathione S-transferases A (GST-A) and A1 (GST-A1) from plaice (Pleuronectes platessa) were expressed as N-terminally 6XHis tagged proteins in Escherichia coli and purified to homogeneity from Ni-NTA silica. GST-A was an efficient catalyst for conjugation of unsaturated alkenals derived from peroxidation of polyunsaturated fatty acids with the highest activity observed with trans-non-2-enal (8 micromol min(-1) mg(-1)). GST-A1 was a very efficient Se-independent glutathione peroxidase with an activity towards cumene hydroperoxide of 25 micromol min(-1) mg(-1). Although the enzymes exhibited moderately high activities towards the model substrate 1-chloro-2,4-dinitrobenzene (CDNB) they exhibited little or no activity towards other common prototypical xenobiotic substrates. Together with data for ontogeny, tissue distribution and inducibility of these enzymes, we contend that a primary function of these enzymes is protection from the harmful effects of lipid peroxidation products generated naturally or exacerbated by xenobiotic exposure.     

Short-term and interannual variability of redox-sensitive chemical parameters in hypoxic/anoxic bottom waters of the Chesapeake Bay

A combination of CTD casts, discrete bottle sampling and in situ voltammetric microelectrode profiling was used to examine changing redox conditions in the water column at a single station south of the Bay Bridge in the upper Chesapeake Bay in late July/early August, 2002–2005. Short-term (2–4 h) fluctuations in the oxic/suboxic/anoxic interface were documented using in situ voltammetric solid-state electrodes. Profiles of dissolved oxygen and sulfide revealed tidally-driven vertical fluctuations of several meters in the depth and thickness of the suboxic zone. Bottom water concentrations of sulfide, Mn²⁺ and Fe²⁺ also varied over the tidal cycle by approximately an order of magnitude. These data indicate that redox species concentrations at this site varied more due to physical processes than biogeochemical processes. Based on analysis of ADCP data, tidal currents at this station were strongly polarized, with the principal axis of tidal currents aligned with the mainstem channel. Together with the chemical data, the ADCP analysis suggests tidal flushing of anoxic bottom waters with suboxic water from north of the site. The present study is thus unique because while most previous studies have focused on processes across relatively stable redox interfaces, our data clearly demonstrate the influence of rapidly changing physical mixing processes on water column redox chemistry.Also noted during the study were interannual differences in maximum bottom water concentrations of sulfide, Mn²⁺ and Fe²⁺. In 2003, for example, heavy spring rains resulted in severe hypoxia/anoxia in June and early July. While reported storm-induced mixing in late July/early August 2003 partially alleviated the low-oxygen conditions, bottom water concentrations of sulfide, Mn²⁺ and Fe²⁺ were still much higher than in the previous year. The latter implies that the response time of the microbial community inhabiting the suboxic/anoxic bottom waters to changing redox conditions is slow compared to the time scale of episodic mixing events. Bottom water concentrations of the redox-sensitive chemical species should thus be useful as a tracer to infer prior hypoxic/anoxic conditions not apparent from ambient oxygen levels at the time of sampling.     

Sedimentology of a subtidal,tide-dominated sand body in the Yellow Sea,southwest Korea

Odanam Satoe, a subtidal, tide-dominated sand body in the Yellow Sea, Korea, is linear in plan and asymmetrical in cross-section. It consists of fine- to medium-grained, well-sorted subangular sand. Bedforms consist of high-amplitude (1–2 m) sandwaves on the lower flanks of the gentler-sloping bar surface, and medium-amplitude (0.5-1 m) sandwaves on the sand body trough adjoining the steeper face, the bar crest and shallower parts of the gently sloping bar surface. Bedforms are absent on the relatively steeper bar surface, which is characterized by 2° slopes. Bedform orientation on the gentler slope is oblique by 30° to the bar crest, parallel to the sand-body crest on the crest itself, and opposite to the steeper sand-body face in the trough below the steeper slope of the bar.Bottom current velocity data show that tidal currents are semi-rotary with a flood time—velocity asymmetry over the gentler slope, and ebb time—velocity asymmetry over the steeper slope during most of the tidal cycle. Tidal-current flow parallels bar elongation over the steeper slope, whereas over the gentler slope, tidal-current flow is directed at 30° to the bar crest and changes to normal to the crest one hour prior to low tide. Bedform orientation mapped with side-scan sonar shows agreement with these flow directions.Sand dispersal around the sand body is controlled by time—velocity asymmetry and partial rotary flow directions of tidal currents. This circulation causes not only a trapezoidal mode of grain dispersal, but also westerly migration of the sand body documented from comparative bathymetric surveys in 1964 and 1980.     

Characterization and molecular analysis of hepatic microsomal UDP-glucuronosyltransferases in the marine fish, Pleuronectes platessa

Here we report the results of the first molecular enzymological study of fish UDP-glucuronosyltransferase (UDPGT). UDPGT activities of plaice liver microsomes were greatest for planar phenols and there was low but measurable conjugation of bilirubin, testosterone and androsterone. A highly purified preparation was isolated which possessed activities towards 1-naphthol, bilirubin and testosterone, containing several molecular weight species of M_r 52–56 kDa. On immunoblot analysis these proteins cross-reacted with a polyspecific anti-rat UDPGT antibody, suggesting that a number of plaice UDPGT isoforms with epitopes in common with the corresponding rat enzymes had been purified.