An untargeted LC-MS metabolomics approach to the metabolic profiles of bottom cultured scallops (Mizuhopecten yessoensis) subjected to mechanical shock in early post-harvest handling
Introduction
Scallops are the main marine shellfish species in economic aquaculture with a top economic value of 2638 $/t in 2017 worldwide (FAO, 2019). The cultivated species Mizuhopecten yessoensis is economically available as a seafood product in China. This species is distributed along the coast of the Dalian Sea in China and is consumed as a live product and is considered a delicacy (Fisheries Bureau of Ministry of Agriculture, China, 2018). During the early post-harvest process, live scallops suffer acute and complex stress induced by temperature fluctuations, improper pressure and salinity, mechanical shock, and aerial exposure (Liu et al., 2017; Yang et al., 2015). These stresses can impact the physiological metabolism of live scallops such as the consumption of glycogen (Takahashi and Mori, 1971) and ATP (Maguire et al., 1999) and the variation in free amino acid content in muscle (Wongso et al., 2007), which can lead to flavor changes and deterioration. Surveys from the main acquisition base of bottom cultured Yesso scallops in China show that mechanical shock is an unavoidable stress factor during harvest and post-harvest handling. In the early mornings, scallops are collected by boat dredges and are unloaded onto boat decks from heights of greater than one meter and are then transported to land-based factories after the empty and dead shells have been manually removed. During the post-harvest process, scallops are subjected to frequent physical stresses, including being stacked in the fishing gear, being dropped to the deck during unloading, and colliding during the sorting and loading processes. Fish welfare is an important emerging concern and is applicable to the harvesting and post-harvest handling processes. Reducing stress has been shown to increase the physical quality, nutritional value and subsequent shelf life of fishery products (Albalat et al., 2010). Stress and mortality for scallops in the supply chain from harvest to market have previously been reported (Woll and Bakke, 2017). However, the direct and indirect impacts of physical stresses on the quality of scallops have not attracted sufficient attention in China. Research on the vitality mechanisms and evaluations of live scallops in early post-harvest handling are inadequate. Conventional evaluation methods such as observations of the withdrawal of the mantle edge in the aquatic industry and measurements of ATP or glycogen in the laboratory are not adequate to fully reflect the physiological status of scallops. Liu et al (Liu et al., 2019) showed that various stresses with acute and complex during early post-harvest handling, which differ from the stresses of the growing environment and can cause physiological stress responses and influence the quality stability of live products in the subsequent purification and circulation. Thus, it is essential to interpret the mechanisms for freshness quality of live scallops under acute stress responses.
Metabolomics is a very effective technique to comprehensively analyze a wide array of metabolites in biological samples and was initially used in disease diagnosis and drug development (German et al., 2005; Nicholson et al., 1999). Metabolomics has also been applied to food science and nutritional research (Wishart, 2008), such as investigations of the organic compounds in milk (Hu et al., 2007), prediction of a biological basis for variations in the quality of chicken meat (Beauclercq et al., 2016), evaluation of quality traits in apple fruits (Cuthbertson et al., 2012), and nutrient metabolites and nonnutrient phytochemicals in edible plants (Ehrman et al., 2008). Several metabolic studies of bivalves have been conducted to indicate marine environmental pollution (Cappello et al., 2013; Hurleysanders et al., 2009; Yusof et al., 2004). Chen et al (Chen et al., 2015) reported a metabolomic investigation of Chlamys farreri exposed to air. However, there has been a lack of research that addresses the metabolic characteristics and quality of scallops under stress during the post-harvest period.
In this paper, the metabolic characteristics of live scallops subjected to mechanical shock were investigated by simulated physical stresses in the early post-harvest process. A metabolomics strategy based on ultra performance liquid chromatography–mass spectrometry (UPLC-MS) was conducted and was followed by a time-series assay of phosphoarginine and ATP-related compounds as indicators of energy consumption of scallops. This study can help to reveal the metabolic profiles of live scallops subjected to mechanical shock during early post-harvest handling and can aid in improving the disposal methods of post-harvest scallops.
Section snippets
Reagents
HPLC grade acetonitrile, methanol and aqueous ammonia were obtained from Merck (Darmstadt, Germany). Nucleotide standards and ammonium acetate were obtained from Sigma-Aldrich (St. Louis, MO, USA). Analytical grade perchloric acid, potassium hydroxide, triethylamine, and phosphoric acid were purchased from the China National Pharmaceutical Group Corporation (Shanghai, China). Ultrapure water (18.2 MΩ) was prepared using a Milli-Q water purification system (Millipore, Shanghai, China).
Scallop samples
The
Phosphoarginine levels
Phosphoarginine is widely used to evaluate the quality of seafood products (Wongso et al., 2007). It is thought to contribute to the energy of aerobic metabolism (Koormann and Grieshaber, 1980; Livingstone et al., 1981).There was no significant difference in phosphoarginine contents between control group and shock group at 0–12 h, However the contents in the shock group fluctuated greatly and exhibited an significant decrease at 36 h (P < 0.05) (shown in Fig. 1).
ATP-related compound levels
ATP-related compounds have been
Conclusion
Here, a metabolomic strategy based on ultra performance liquid chromatography–mass spectrometry (UPLC-MS) and multivariate data analysis was used to investigate the metabolic characteristics of live scallops under physical stress during early post-harvest handling. As a result, 18 biomarkers were identified, and their levels differed significantly between the control and shock groups. The results suggested clear metabolic disturbances in scallops subjected to abrupt mechanical shock, which are
Declaration of Competing Interest
The authors declare no conflict of interest.
Acknowledgements
This work received financial support from National Key R&D Program of China (NO. 2018YFD0901001) and National Natural Science Foundation of China (NO. 31671790).
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