Use of suppressive subtractive hybridization and cDNA arrays to discover patterns of altered gene expression in the liver of dihydrotestosterone and 11-ketotestosterone exposed adult male largemouth bass (Micropterus salmoides)

In this study male largemouth bass (LMB) were exposed to the naturally occurring androgens, dihydrotestosterone (DHT) or 11-ketotestosterone (11-KT) in order to identify genes that are differentially regulated by these steroid hormones. Using subtractive hybridization on livers of fish treated with DHT against vehicle control, many novel LMB genes were cloned. These genes were added to our gene library and arrayed. Six genes were up-regulated and five were down-regulated by both androgens. But, each androgen also regulated specific genes. One gene that was identified as a potential androgen marker was spermidine-spermine-N(1)-acetyltransferase that was up-regulated by both androgens. Determining which genes are responsive to natural androgens will help to identify biochemical pathways that are impacted.     

Contaminant concentrations and toxicity of sea-surface microlayer near Los Angeles, California

Sea-surface microlayer samples were collected from six nearshore areas receiving different amounts of anthropogenic inputs. The samples were analyzed for selected trace metals, chlorinated hydrocarbons, and polycyclic aromatic hydrocarbons. The relative toxicities of the samples were determined with fish embryo bioassays.Contaminant concentrations generally increased from offshore to the inshore stations. Contaminant concentrations were several orders of magnitude higher in microlayer samples from the highly industrialized Los Angeles and Long Beach harbors compared to samples from a site 15km offshore. Microlayer samples from the inshore stations were significantly more toxic, and induced significantly more developmental abnormalities and chromosome aberrations, than samples from the offshore stations.     

Swath mapping of the seafloor and its application to deep-bottom fisheries in New Caledonia

A bottom longline fishery operated in the EEZ of New Caledonia from 1988 to 1991. Fishing focused on five seamounts with summits at depths ranging from 500 to 750 m. The target species was alfonsino, Beryx splendens. As the soundings available from marine charts were not detailed enough, the fishing masters had to make their own charts in order to set the gear in the right location. A series of 11 scientific cruises devoted to a research program on alfonsino was launched in late 1991. During the first scientific cruises, several days were spent mapping the seabed to improve the existing knowledge of the topography of three seamounts by coupling the EDO echosounder depth measurements to the GPS positions. As this procedure is slow, it is applicable to limited areas or, if a wider grid is used, to large zones to locate major structures such as ridges and seamounts. The emergence of the multibeam echosounder has greatly improved seabed mapping performance. This tool seems to fit the requirement of exploratory deep-bottom fishing particularly well, as it covers large areas while providing details of the bottom. The EM 12 was used to obtain bathymetry and imagery of the SE portion of the EEZ of New Caledonia, covering an approximate area of 70,000 km² in two weeks. For depths ranging from 500 to 4,500 m, the results were impressive. They confirmed the known major features and provided greater topographical detail, revealing fine unknown structures. They also provided information on the type of substratum, information which might influence the fishing strategy. Finally, they made it possible to obtain an accurate estimate of the exploitable area which in turn led to new stock estimates.     

Cloning of a chub metallothionein cDNA and development of competitive RT-PCR of chub metallothionein mRNA as a potential biomarker of heavy metal exposure

Metallothionein has been assayed in a range of aquatic animal tissues as an indicator of metal exposure. We sequenced chub (Leuciscus cephalus) metallothionein cDNA which showed over 90% homology to common carp, goldfish and stone loach and 77% homology to rainbow trout sequences for metallothionein. We then used the extended primer method to develop an accurate quantitative competitive RT-PCR assay for metallothionein mRNA. RT-PCR was used to measure metallothionein mRNA in feral chub from a range of field sites, with different levels of heavy metal pollution, in the West Midlands, UK. Measurements were complemented by analysis of liver and gill metallothionein protein by capillary electrophoresis. There was no significant difference in the metallothionein protein levels between fish of different rivers and there was no evidence of elevation of mRNA at the sites of highest metal exposure. The level of metal exposure (e.g. zinc, nickel and cadmium each ranging between 15 and 28 microg/l ) at the pH (7.5-8.5) of these rivers appears insufficient to elevate hepatic or gill metallothionein in chub. A lack of elevation of hepatic metallothionein mRNA in chub exposed to zinc, copper and manganese for 24 h and 10 days in the laboratory also suggests a non-responsiveness of this species.     

Respiratory Rate and Assessment of Secondary Production in the Holothuroid Holothuria tubulosa (Echinodermata) from Mediterranean Seagrass Beds

Abstract. Respiration in Holothuria tubulosa was investigated in individuals from the Posidonia oceanica meadow off Lacco Ameno (Ischia Island, Italy). Respiratory rates increase with increasing body weight and increasing sea water temperature. Oxygen consumption of an average individual (7g dw body wall) ranges from 0.409 (14 °C) to 1.300 (26 °C) mg O₂· h^-1. Data on population density, mean size of individuals, and annual sea water temperature variations allow an assessment of holothuroid production. Values of 45.65 and 13.75 kJ · m^-2· y^-1 were calculated for shallow (3 to 10 m) and deep (25 to 33 m) areas of the Posidonia meadow, respectively. Holothuroid production shows a bathymetric pattern similar to primary production of the Posidonia -epiphytes complex and the production of Posidonia litter.     

Nonparametric Estimates of the Sea State Bias for the Jason-1 Radar Altimeter

The Jason-1 sea state bias (SSB) is analyzed in depth from the first year of GDR products. Compared to previous missions, this work benefits from two aspects of the empirical determination of the SSB from the altimetric data themselves. First, from a methodological point of view, a nonparametric technique (NP) has been developed and largely tested on TOPEX/Poseidon 1, GFO and Envisat data. The NP estimator has proven to be a useful tool in the SSB estimation, and it is now mature enough to be used for a refined analysis. On the other hand, the SSB can be extracted from three different data sets (crossovers, collinear data, and residuals) with different characteristics. It is then possible to cross calibrate various estimations of the SSB models and to determine the most accurate one. A systematic comparison is made between these different estimates for the Jason-1 altimeter. The collinear and crossover data sets yield very similar estimates despite their difference of spatial and temporal distributions. These SSB models assure consistency with the TOPEX mission when comparing Jason-1 and TOPEX residuals during the tandem phase. Thanks to the present work, the impact of the short wavelengths filtering on the SSB estimation is evidenced. More generally, our understanding of potential errors affecting the sea surface height and their impact onto the SSB estimation is also improved.     

Bioenhancement of cadmium transfer along a multi-level food chain

Previous studies have shown that metal partitioned to a subcellular compartment containing trophically available metal (TAM) is readily available to predators and may be enhanced by increased binding of metal to heat-stable proteins (HSP - e.g., metallothioneins). The aim of the current investigation was to determine the influence of TAM on the trophic transfer of Cd along an experimental, three-level food chain: Artemia franciscana (brine shrimp)-->Palaemonetes pugio (grass shrimp)-->Fundulus heteroclitus (mummichog). P. pugio were fed for 7 days on A. franciscana exposed to Cd in solution (including (109)Cd as radiotracer) and subjected to subcellular fractionation or fed to F. heteroclitus. An HSP-driven increase in the percentage of Cd associated with TAM (TAM-Cd%) in A. franciscana exposed to 1 muM Cd resulted in a bioenhancement (i.e., a greater than linear increase with respect to A. franciscana exposure) of Cd trophic transfer to P. pugio. Increased dietary Cd exposure did not affect TAM-Cd% in P. pugio nor trophic transfer to F. heteroclitus.     

New age constraints on emplacement of the Cévenol granitoids, South French Massif Central

During the development of the Variscan orogeny, large amounts of granitic melt were produced, giving rise to the intrusion of granitoids at different structural levels. Despite numerous studies, ages available from previous work on the Cévennes granites remain largely imprecise. In order to better constrain the age and emplacement mode of these granites, we have combined U–Pb dating on monazites and zircons and ⁴⁰Ar/³⁹Ar dating on biotites with petrological observations, major element chemical analysis and SEM zircon imaging on five samples from the Aigoual–St Guiral–Liron and Mont Lozère granitic massifs. The results revealed that granitic intrusions and cooling in Southern Cévennes occurred in a short time span at ∼306 Ma after the main episode of regional metamorphism. Petrological and chemical data suggest that they result from a mixing between mantle-derived basic magmas (lamprophyres) and lower crust acid magmas. At a regional scale the production of these melts occurred at the end of crustal thickening induced by nappe stacking, at the same time as the late anatectic events recorded further north in the Velay dome and the granulite facies metamorphism recorded in metasedimentary granulite enclaves brought up by Tertiary volcanoes of the Velay area (Bournac).     

Nutrient Mass Flux between Florida Bay and the Florida Keys National Marine Sanctuary

There is a net discharge of water and nutrients through Long Key Channel from Florida Bay to the Florida Keys National Marine Sanctuary (FKNMS). There has been speculation that this water and its constituents may be contributing to the loss of coral cover on the Florida Keys Reef tract over the past few decades, as well as speculation that changes in freshwater flow in the upstream Everglades ecosystem associated with the Comprehensive Everglades Restoration Plan may exacerbate this phenomenon. The results of this study indicate that although there is a net export of approximately 3,850 (±404) ton N year^?1 and 63 (±7) ton P year^?1, the concentrations of these nutrients flowing out of Florida Bay are the same as those flowing in. This implies that no significant nutrient enrichment is occurring in the waters of the FKNMS in the vicinity of Long Key Channel. Because of the effect of restricted southwestward water flow through Florida Bay by shallow banks and small islands, the volume of relatively high-nutrient water from central and eastern portions of the bay exiting through the channel is small compared to the average tidal exchange. Nutrient loading of relatively enriched bay waters is mediated by tidal exchange and mixing with more ambient concentrations of the western Florida Bay and Hawk Channel. System-wide budgets indicate that the contribution of Florida Bay waters to the inorganic nitrogen pool of the Keys coral reef is small relative to offshore inputs.