Awanis Azizan1,2, Andrea C Alfaro3,4, Tim Young1,5,2, Leonie Venter1,2. 1. Aquaculture Biotechnology Research Group, Department of Applied Ecology, School of Science, Auckland University of Technology, Private Bag 92006, Auckland, 1142, New Zealand. 2. Faculty of Health and Environmental Sciences, Institute for Applied Ecology New Zealand, School of Applied Sciences, Auckland University of Technology, Private Bag 92006, Auckland, 1142, New Zealand. 3. Aquaculture Biotechnology Research Group, Department of Applied Ecology, School of Science, Auckland University of Technology, Private Bag 92006, Auckland, 1142, New Zealand. andrea.alfaro@aut.ac.nz. 4. Faculty of Health and Environmental Sciences, Institute for Applied Ecology New Zealand, School of Applied Sciences, Auckland University of Technology, Private Bag 92006, Auckland, 1142, New Zealand. andrea.alfaro@aut.ac.nz. 5. Centre for Biomedical & Chemical Sciences, School of Science, Auckland University of Technology, Auckland, New Zealand.
Abstract
BACKGROUND: The New Zealand Green-lipped mussel industry is well-established providing vastly to aquaculture exports. To assess mussel health and reproduction status, visual examination of organs and/or collection of haemolymph is commonly applied. Anesthetics, such as magnesium chloride (MgCl2) can be utilized to prevent muscle contraction and keep shells open during sampling. The specific effects of muscle relaxing agents on baseline metabolism in invertebrates is unknown, but it is evident that molecular, cellular and physiological parameters are altered with these chemical applications. To this end, metabolomics approaches can help elucidate the effects of relaxing agents for better assessment of their use as a research tool. METHODS: Adult Green-lipped mussels were anaesthetized for 3 h in a MgCl2 bath, whereafter haemolymph samples were collected and analyzed via gas chromatography-mass spectrometry applying methyl chloroformate alkylation derivatization. RESULTS: Anesthetized mussels were characterized as non-responsive to manual manipulation, with open valves, and limited siphoning function. Metabolite profiling revealed significant increases in the abundances of most metabolites with an array of metabolic activities affected, resulting in an energy imbalance driven by anaerobic metabolism with altered amino acids acting as neurotransmitters and osmolytes. CONCLUSION: This research is the first to use a metabolomics approach to identify the metabolic consequences of this commonly used bivalve relaxing technique. Ultimately the use of MgCl2 anesthetization as a sampling strategy should be carefully evaluated and managed when performing metabolomics-related research.
BACKGROUND: The New Zealand Green-lipped mussel industry is well-established providing vastly to aquaculture exports. To assess mussel health and reproduction status, visual examination of organs and/or collection of haemolymph is commonly applied. Anesthetics, such as magnesium chloride (MgCl2) can be utilized to prevent muscle contraction and keep shells open during sampling. The specific effects of muscle relaxing agents on baseline metabolism in invertebrates is unknown, but it is evident that molecular, cellular and physiological parameters are altered with these chemical applications. To this end, metabolomics approaches can help elucidate the effects of relaxing agents for better assessment of their use as a research tool. METHODS: Adult Green-lipped mussels were anaesthetized for 3 h in a MgCl2 bath, whereafter haemolymph samples were collected and analyzed via gas chromatography-mass spectrometry applying methyl chloroformate alkylation derivatization. RESULTS: Anesthetized mussels were characterized as non-responsive to manual manipulation, with open valves, and limited siphoning function. Metabolite profiling revealed significant increases in the abundances of most metabolites with an array of metabolic activities affected, resulting in an energy imbalance driven by anaerobic metabolism with altered amino acids acting as neurotransmitters and osmolytes. CONCLUSION: This research is the first to use a metabolomics approach to identify the metabolic consequences of this commonly used bivalve relaxing technique. Ultimately the use of MgCl2 anesthetization as a sampling strategy should be carefully evaluated and managed when performing metabolomics-related research.
Authors: Sili Fan; Tobias Kind; Tomas Cajka; Stanley L Hazen; W H Wilson Tang; Rima Kaddurah-Daouk; Marguerite R Irvin; Donna K Arnett; Dinesh K Barupal; Oliver Fiehn Journal: Anal Chem Date: 2019-02-19 Impact factor: 6.986
Authors: Halina I Falfushynska; Lesya L Gnatyshyna; Olesya Y Osadchuk; Anna Farkas; Agnes Vehovszky; David O Carpenter; Janos Gyori; Oksana B Stoliar Journal: Comp Biochem Physiol C Toxicol Pharmacol Date: 2014-05-09 Impact factor: 3.228