Dead leaves of the Neptune grass, Posidonia oceanica (L.) Delile, in the Mediterranean coastal zone, are colonized by an abundant "detritivorous" invertebrate community that is heavily predated by fishes. This community was sampled in August 2011, November 2011, and March 2012 at two different sites in the Calvi Bay (Corsica). Ingested artificial fibers (AFs) of various sizes and colors were found in 27.6% of the digestive tracts of the nine dominant species regardless of their trophic level or taxon. No seasonal, spatial, size, or species-specific significant differences were revealed; suggesting that invertebrates ingest AFs at constant rates. Results showed that, in the gut contents of invertebrates, varying by trophic level, and across trophic levels, the overall ingestion of AFs was low (approximately 1 fiber per organism). Raman spectroscopy revealed that the ingested AFs were composed of viscose, an artificial, cellulose-based polymer. Most of these AFs also appeared to have been colored by industrial dyes. Two dyes were identified: Direct Blue 22 and Direct Red 28. The latter is known for being carcinogenic for vertebrates, potentially causing environmental problems for the P. oceanica litter community. Techniques such as Raman spectroscopy are necessary to investigate the particles composition, instead of relying on fragment size or color to identify the particles ingested by animals.

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Modern ion mobility instrumentation is typically operated above the low field limit, which may activate the ions and cause structural rearrangement or fragmentation during analysis. Here, we quantitatively assessed the internal heating experienced by ions during trapped ion mobility spectrometry (TIMS) experiments. To this end, the fragmentation yields of fragile benzylpyridinium “thermometer” ions were monitored during both the accumulation and analysis steps inside the TIMS tunnel. The corresponding fragmentation rate constants were translated into a vibrational effective temperature T eff,vib. Our results demonstrate significant fragmentation upstream and inside the TIMS tunnel that corresponds to T eff,vib ≈ 510 K during both the accumulation and analysis steps. Broadening our scope to cytochrome c and lysozyme, we showed that although compact “native” folds can be preserved, the collision cross section distributions are highly sensitive to the transmission voltages and the analysis time scale. Our results are discussed with regard to T eff,vib data previously acquired on traveling-wave (TWIMS) ion mobility in the context of native mass spectrometry and conformational landscape exploration.

Microplastic particles (MP) contaminate oceans and affect marine organisms in several ways. Ingestion combined with food intake is generally reported. However, data interpretation often is circumvented by the difficulty to separate MP from bulk samples. Visual examination often is used as one or the only step to sort these particles. However, color, size, and shape are insufficient and often unreliable criteria. We present an extraction method based on hypochlorite digestion and isolation of MP from the membrane by sonication. The protocol is especially well adapted to a subsequent analysis by Raman spectroscopy. The method avoids fluorescence problems, allowing better identification of anthropogenic particles (AP) from stomach contents of fish by Raman spectroscopy. It was developed with commercial samples of microplastics and cotton along with stomach contents from three different Clupeiformes fishes: Clupea harengus, Sardina pilchardus, and Engraulis encrasicolus. The optimized digestion and isolation protocol showed no visible impact on microplastics and cotton particles while the Raman spectroscopic spectrum allowed the precise identification of microplastics and textile fibers. Thirty-five particles were isolated from nine fish stomach contents. Raman analysis has confirmed 11 microplastics and 13 fibers mainly made of cellulose or lignin. Some particles were not completely identified but contained artificial dyes. The novel approach developed in this manuscript should help to assess the presence, quantity, and composition of AP in planktivorous fish stomachs.

SUMMARYIsocitrate lyase is a key enzyme of the glyoxylate cycle. This cycle plays an essential role in cell growth on acetate, and is important for gluconeogenesis as it bypasses the two oxidative steps of the tricarboxylic acid (TCA) cycle in which CO 2 is evolved. In this paper, a null icl mutant of the green microalga Chlamydomonas reinhardtii is described. Our data show that isocitrate lyase is required for growth in darkness on acetate (heterotrophic conditions), as well as for efficient growth in the light when acetate is supplied (mixotrophic conditions). Under these latter conditions, reduced acetate assimilation and concomitant reduced respiration occur, and biomass composition analysis reveals an increase in total fatty acid content, including neutral lipids and free fatty acids. Quantitative proteomic analysis by 14 N/ 15 N labelling was performed, and more than 1600 proteins were identified. These analyses reveal a strong decrease in the amounts of enzymes of the glyoxylate cycle and gluconeogenesis in parallel with a shift of the TCA cycle towards amino acid synthesis, accompanied by an increase in free amino acids. The decrease of the glyoxylate cycle and gluconeogenesis, as well as the decrease in enzymes involved in b-oxidation of fatty acids in the icl mutant are probably major factors that contribute to remodelling of lipids in the icl mutant. These modifications are probably responsible for the elevation of the response to oxidative stress, with significantly augmented levels and activities of superoxide dismutase and ascorbate peroxidase, and increased resistance to paraquat.

The National Institute of Standards and Technology (NIST), in collaboration with the National Institutes of Health (NIH), has developed a Standard Reference Material (SRM) to support technology development in metabolomics research. SRM 1950 Metabolites in Human Plasma is intended to have metabolite concentrations that are representative of those found in adult human plasma. The plasma used in the preparation of SRM 1950 was collected from both male and female donors, and donor ethnicity targets were selected based upon the ethnic makeup of the U.S. population. Metabolomics research is diverse in terms of both instrumentation and scientific goals. This SRM was designed to apply broadly to the field, not toward specific applications. Therefore, concentrations of approximately 100 analytes, including amino acids, fatty acids, trace elements, vitamins, hormones, selenoproteins, clinical markers, and perfluorinated compounds (PFCs), were determined. Value assignment measurements were performed by NIST and the Centers for Disease Control and Prevention (CDC). SRM 1950 is the first reference material developed specifically for metabolomics research.

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Anthropogenic particles (APs) are a very broad category of particles produced directly or indirectly by human activities. Their ingestion by biota is well studied in the marine environment. In contrast, studies on AP ingestion in wild freshwater organisms are scarce despite high contamination levels in some rivers and lakes. In this study, we aimed to evaluate the ingestion of APs and the possible occurrence of APs in the liver and muscle of a freshwater fish, Squalius cephalus, from the Parisian conurbation. After isolation, the particles were analyzed using Raman spectroscopy. In sixty stomachs, eighteen APs were found, half of which were plastics and the other half were dyed particles. Twenty-five percent of sampled individuals had ingested at least one AP. The mean length of the APs was 2.41 mm. No significant difference was found between the sites upstream and downstream of Paris. Additionally, 5% of sampled livers contained one or more APs, which were characterized as microplastics (MPs). No APs were found in the muscle tissue. The majority of APs isolated from stomach contents were fibers, which is similar to the findings of a previous river contamination study. This highlights that fish could be more exposed to fibers than previously thought and that more studies on the impacts of fiber ingestion are required. Despite their low occurrence, MPs are reported, for the first time, in the liver of a wild freshwater fish species. While the pathways and impacts are still unknown, MPs also occur in liver of marine mollusks and fish. Physiological in vitro studies are needed to better evaluate the impacts of such phenomena.