A selection of reference genes and early-warning mRNA biomarkers for environmental monitoring using Mytilus spp. as sentinel species

International audiencemRNA biomarkers are promising tools for environmental health assessment and reference genes are needed to perform relevant qPCR analyses in tissue samples of sentinel species. In the present study, potential reference genes and mRNA biomarkers were tested in the gills and digestive glands of native and caged mussels (Mytilus spp.) exposed to harbor pollution. Results highlighted the difficulty to find stable reference genes in wild, non-model species and suggested the use of normalization indices instead of single genes as they exhibit a higher stability. Several target genes were found differentially expressed between mussel groups, especially in gills where cyp32, pi-gst and CuZn-sod mRNA levels could be biomarker candidates. Multivariate analyses confirmed the ability of mRNA levels to highlight site-effects and suggested the use of several combined markers instead of individual ones. These findings support the use of qPCR technology and mRNA levels as early-warning biomarkers in marine monitoring programs

Development of an innovative and “green” stir bar sorptive extraction–thermal desorption–gas chromatography–tandem mass spectrometry method for quantification of polycyclic aromatic hydrocarbons in marine biota

International audienceThere is a growing awareness of the need to reduce the negative impact of chemical analyses on the environment and to develop new eco-friendly and sustainable analytical methods without compromising performance. In this study, we developed a "green" analytical method enabling the accurate and simultaneous routine analysis of 21 polycyclic aromatic hydrocarbons (PAHs) in reduced quantities (100 mg and 1 g wet weight (WW)) of marine biota samples (fish muscle, mussel and oyster tissues) using alkaline digestion combined with stir bar sorptive extraction-thermal desorption-gas chromatography-tandem mass spectrometry (SBSE-GC-MS/MS). The innovative method provides good selectivity and specificity for most compounds. In 1 gWW samples, limits of quantification (LOQs) ranged from 1 to 10 mu g/kgWW in fish muscle and from 0.5 to 10 mu g/kgWW in mussel tissue. The method enables most analytes to be quantified below the restrictive limits established by the European Commission (2 and 10 mu g/kgWW in fish muscle and bivalve mollusc, respectively). Higher LOQs were obtained in 100 mgWW samples ranging from 1 to 50 mu g/kgWW. Recovery and linearity were assessed for all analytes. The results were satisfactory for most compounds with recoveries ranging from 94% to 117% in 1 gWW mussel samples at spike concentration of 10 ng/gWW with standard deviation not exceeding 12%. However, results confirmed that the SBSE efficiency is affected by the complexity of biological matrices, especially for high molecular weight compounds in lipid-rich mussel tissue. Because of the matrix effects, matrix-matched calibrations were carried out. Validation was performed using the standard reference material 1974c with recovery ranging from 71% to 119% except for naphthalene, anthracene and benzo(e)pyrene that were therefore not validated. Overall, the developed method meets analytical validation criteria for most compounds. Thanks to the combination of alkaline digestion and SBSE, which greatly simplifies sample treatment and limits solvent use to ethanol, the developed method followed most green analytical chemistry principles

Seasonal monitoring of blue mussel ( Mytilus spp. ) populations in a harbor area: A focus on responses to environmental factors and chronic contamination

International audienceCoastal waters corresponding to macrotidal systems are among the most variable marine biotopes. Sessile animals as bivalve mollusks may however be found forming intertidal beds at high densities, as allowed by full adaptation to local conditions. A better knowledge of adaptive responses to environmental factors is required to foresee possible adverse effects of global change. At the sub-cellular level, transcriptional responses are among the earliest signals of environmental disturbances and they can reveal subtle and meaningful changes in organism exposed to stress. Three blue mussel (Mytilus spp.) populations inhabiting the Bay of Brest (France) in sites exposed to different levels of chronic pollution, from low to moderate, were surveyed upon a seasonal schedule, with special attention to the reproductive cycle. Major seawater parameters were monitored over a full-year in the framework of the S! RANO project, based on an automatic high frequency acquisition system installed aboard a ship of opportunity. The health status of mussels has been assessed by measuring a condition index and gametogenesis has been followed by histology. Selected biological responses to environmental stress were detected using a multimarker approach including expression of genes involved in chemical stress response and energetic metabolism, and cellular immune parameters. Environmental parameters showed deep seasonal variations which differed among sites. Most biological responses followed a seasonal pattern. Late winter and spring corresponded to an active reproduction period in the Bay of Brest. Earlier spawning was observed in harbor areas compared to the oceanic site and an altered physiological state was assumed in commercial harbor mussels during the reproductive period, suggesting that their health is compromised at this time of year. However, no signs of severe chemical stress were detected in both harbor mussel populations, which could reflect adaptive responses to adverse environmental conditions

High diversity of oilseed rape pollen clouds over an agro-ecosystem indicates long-distance dispersal

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Development of a high-power THz radiation source for plasma diagnostics

A high power radiation source in the THz range with long pulse and narrow line width is required for diagnosing fusion type plasmas by collective Thomson scattering. Gyrotrons currently meet the requirements concerning power, pulse length and line width when operating in the 50-150 GHz range. They may have the potential also to do this in the THz range. In this paper two methods based on harmonic operation are investigated. Both look promising, if high conversion efficiency can be attained. This, however, still has to be demonstrated. (C) 1999 Published by Elsevier Science B.V. All rights reserved