title: Haemolymph from Mytilus galloprovincialis: Response to copper and temperature challenges studied by (1)H-NMR metabonomics.
abstract: Numerous studies on molluscs have been carried out to clarify the physiological roles of haemolymph serum proteins and haemocytes. However, little is known about the presence and functional role of the serum metabolites. In this study, Nuclear Magnetic Resonance (NMR) was used to assess whether changes of the metabolic profile of Mytilus galloprovincialis haemolymph may reflect alterations of the physiological status of the organisms due to environmental stressors, namely copper and temperature. Mussel haemolymph was taken from the posterior adductor muscle after a 4-day exposure to ambient (16 °C) or high temperature (24 °C) and in the absence or presence (5 μg/L, 20 μg/L, or 40 μg/L) of sublethal copper (Cu(2+)). The total glutathione (GSH) concentration in the haemolymph of both control and treated mussels was minimal, indicating the absence of significant contaminations by muscle intracellular metabolites due to the sampling procedure. In the (1)H-NMR spectrum of haemolymph, 27 metabolites were identified unambiguously. The separate and combined effects of exposure to copper and temperature on the haemolymph metabolic profile were assessed by Principal Component Analysis (PCA) and Ranking-PCA multivariate analysis. Changes of the metabolomic profile due to copper exposure at 16 °C became detectable at a dose of 20 μg/L copper. Alanine, lysine, serine, glutamine, glycogen, glucose and protein aliphatics played a major role in the classification of the metabolic changes according to the level of copper exposition. High temperature (24 °C) and high copper levels caused a coherent increase of a common set of metabolites (mostly glucose, serine, and lysine), indicating that the metabolic impairment due to high temperature is enforced by the presence of copper. Overall, the results demonstrate that, as for human blood plasma, the analysis of haemolymph metabolites represents a promising tool for the diagnosis of pollutant-induced stress syndrome in marine mussels.
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "Amedeo Avogadro", Viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Digilio
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "Amedeo Avogadro", Viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Susanna
authors.initials: S
authors.lastname: Sforzini
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "Amedeo Avogadro", Viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Cassino
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "Amedeo Avogadro", Viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Robotti
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "Amedeo Avogadro", Viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Oliveri
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "Amedeo Avogadro", Viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Emilio
authors.initials: E
authors.lastname: Marengo
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "Amedeo Avogadro", Viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Musso
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "Amedeo Avogadro", Viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Osella
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "Amedeo Avogadro", Viale T. Michel 11, 15121 Alessandria, Italy. Electronic address: viarengo@uniupo.it.
authors.forename: Aldo
authors.initials: A
authors.lastname: Viarengo
mesh_terms: D000818:Animals; D017756:Aquaculture; D015415:Biomarkers / Q000378:metabolism; D003300:Copper / Q000008:administration & dosage / Q000506:poisoning*; D004305:Dose-Response Relationship, Drug; D005260:Female; D005947:Glucose / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D006458:Hemolymph / Q000187:drug effects* / Q000378:metabolism; D006358:Hot Temperature / Q000009:adverse effects; D007558:Italy; D008239:Lysine / Q000378:metabolism; D055432:Metabolomics / Q000379:methods; D049878:Mytilus / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism / Q000502:physiology; D019906:Nuclear Magnetic Resonance, Biomolecular; D025341:Principal Component Analysis; D012694:Serine / Q000378:metabolism; D013312:Stress, Physiological / Q000187:drug effects*; D014018:Tissue Distribution; D066007:Toxicokinetics; D015854:Up-Regulation / Q000187:drug effects; D014874:Water Pollutants, Chemical / Q000008:administration & dosage / Q000506:poisoning*
chemical_list: D015415:Biomarkers; D014874:Water Pollutants, Chemical; D012694:Serine; D003300:Copper; D005978:Glutathione; D005947:Glucose; D008239:Lysine
keywords: (1)H-NMR; Copper; Ecotoxicology; Haemolymph; Metabonomics; Mytilus galloprovincialis; Temperature