Identification of very long-chain (>C24) fatty acid methyl esters using gas chromatography coupled to quadrupole/time-of-flight mass spectrometry with atmospheric pressure chemical ionization source.

Gas chromatography coupled to a quadrupole time-of-flight mass analyzer (QTOF) with an atmospheric pressure chemical ionization (APCI) source has been tested to study the ionization and mass spectrometric behavior of long-chain and very long-chain polyunsaturated fatty acids (LC-PUFAs, C18-24; VLC-PUFAs, >C24). The protonated molecule ([M+H]+), measured at accurate mass, became the base peak of the spectrum for all the studied compounds and was promoted by the addition of water into the source. This fact overcame the existing difficulties for the identification of VLC-PUFAs when using an electron ionization source (EI). The extensive fragmentation of PUFAs in this source is the main drawback due to the fact that since reference standards are not commercially available, final identification relies on retention time estimation. The application of GC-APCI-QTOF to the screening of lipid extracts from the eyes of different fish species added confidence to the identification of several VLC-PUFAs. Further investigation of ion ratios allowed to predict the position of key double bonds enabling the classification of VLC-PUFAs as ω3 or ω6 compounds. VLC-PUFAs spectra found in biological samples were compared to those obtained from corresponding peaks found in heterologous expression experiments of fish's Elovl4.

Effects of ibuprofen and carbamazepine on the ion transport system and fatty acid metabolism of temperature conditioned juveniles of Solea senegalensis.

The increasing presence of pharmaceuticals in aquatic environments in the last decades, derived from human and veterinary use, has become an important environmental problem. Previous studies have shown that ibuprofen (IB) and carbamazepine (CBZ) modify physiological and biochemical processes in Senegalese sole (Solea senegalensis) in a temperature-dependent manner. In other vertebrates, there is evidence that both of these pharmaceuticals interfere with the 'arachidonic acid (AA) cascade', which is responsible for the biosynthesis of numerous enzymes that are involved in the osmoregulatory process. The present work aims to study the temperature-dependent effects of these two pharmaceuticals on several biochemical and molecular parameters in Senegalese sole. Regarding osmoregulation, Na+, K+ -ATPase enzyme activity was determined in the gills, kidney and intestine, and the expressions of both Na+, K+ -ATPase 1α-subunit isoforms (ATP1A1a and ATP1A1b) were quantified in gills. Gill prostaglandin-endoperoxide synthase-2 (PTGS2) gene expression and fatty acid composition were selected to determine the interference of both pharmaceuticals with the AA cascade. Senegalese sole juveniles, acclimatised at 15°C or 20°C, were exposed through intraperitoneal injection to IB (10mg/kg) and CBZ (1mg/kg) for 48h. Non-injected fish (Control) and those injected with the carrier (sunflower oil; S.O.), acclimated at each of the two temperatures, were used for comparison. The results show that IB directly affected the osmoregulatory mechanisms that alter gill and intestine Na+, K+ -ATPase activities. In addition, the copy number of ATP1A1a was higher at 20°C than at 15°C, which could be a direct response to the temperature variation. The gene expression of PTGS2 was affected by neither drug administration nor acclimation temperature. Nevertheless, detailed analysis of AA and eicosapentaenoic acid (EPA) percentages revealed a CBZ-derived effect in the fatty acid composition of the gills.

Functional rare males in diploid parthenogenetic Artemia.

Functional males that are produced occasionally in some asexual taxa - called 'rare males' - raise considerable evolutionary interest, as they might be involved in the origin of new parthenogenetic lineages. Diploid parthenogenetic Artemia produce rare males, which may retain the ability to mate with females of related sexual lineages. Here, we (i) describe the frequency of male progeny in populations of diploid parthenogenetic Artemia, (ii) characterize rare males morphologically, (iii) assess their reproductive role, using cross-mating experiments with sexual females of related species from Central Asia and characterize the F1 hybrid offspring viability and (iv) confirm genetically both the identity and functionality of rare males using DNA barcoding and microsatellite loci. Our result suggests that these males may have an evolutionary role through genetic exchange with related sexual species and that diploid parthenogenetic Artemia is a good model system to investigate the evolutionary transitions between sexual species and parthenogenetic strains.