Department of Ocean Sciences, Memorial University of Newfoundland, 0 Marine Lab Road, St. John's, NL, A1C 5S7, Canada.
National Center for Cool and Cold Water Aquaculture, ARS/USDA, 11861 Leetown Rd, Kearneysville, WV, 25430, USA.
Mar Biotechnol (NY). 2020 Aug;22(4):511-525. doi: 10.1007/s10126-020-09972-5. Epub 2020 Jun 4.
The simultaneous quantification of several transcripts via multiplex PCR can accelerate research in fish physiological responses to diet and enable the development of superior aquafeeds for farmed fish. We designed two multiplex PCR panels that included assays for 40 biomarker genes representing key aspects of fish physiology (growth, metabolism, oxidative stress, and inflammation) and 3 normalizer genes. We used both panels to assess the physiological effects of replacing fish meal and fish oil by terrestrial alternatives on Atlantic salmon smolts. In a 14-week trial, we tested three diets based on marine ingredients (MAR), animal by-products and vegetable oil (ABP), and plant protein and vegetable oil (VEG). Dietary treatments affected the expression of genes involved in hepatic glucose and lipid metabolism (e.g., srebp1, elovl2), cell redox status (e.g., txna, prdx1b), and inflammation (e.g., pgds, 5loxa). At the multivariate level, gene expression profiles were more divergent between fish fed the marine and terrestrial diets (MAR vs. ABP/VEG) than between the two terrestrial diets (ABP vs. VEG). Liver ARA was inversely related to glucose metabolism (gck)- and growth (igfbp-5b1, htra1b)-related biomarkers and hepatosomatic index. Liver DHA and EPA levels correlated negatively with elovl2, whereas ARA levels correlated positively with fadsd5. Lower hepatic EPA/ARA in ABP-fed fish correlated with the increased expression of biomarkers related to mitochondrial function (fabp3a), oxidative stress (txna, prdx1b), and inflammation (pgds, 5loxa). The analysis of hepatic biomarker gene expression via multiplex PCR revealed potential physiological impacts and nutrient-gene interactions in Atlantic salmon fed lower levels of marine-sourced nutrients.
通过多重聚合酶链反应(PCR)同时定量多个转录本可以加速鱼类对饮食的生理反应研究,并为养殖鱼类开发更优质的水产饲料。我们设计了两个多重 PCR 面板,其中包括 40 个生物标志物基因的检测,这些基因代表鱼类生理学的关键方面(生长、代谢、氧化应激和炎症)和 3 个内参基因。我们使用这两个面板来评估用陆地替代品替代鱼粉和鱼油对大西洋鲑幼鱼的生理影响。在一项为期 14 周的试验中,我们测试了三种基于海洋成分(MAR)、动物副产品和植物油(ABP)以及植物蛋白和植物油(VEG)的饮食。饮食处理影响了参与肝葡萄糖和脂质代谢(如 srebp1、elovl2)、细胞氧化还原状态(如 txna、prdx1b)和炎症(如 pgds、5loxa)的基因表达。在多变量水平上,与海洋和陆地饮食相比(MAR 与 ABP/VEG),不同饲料组之间的基因表达谱差异更大(ABP 与 VEG)。肝 ARA 与葡萄糖代谢(gck)和生长(igfbp-5b1、htra1b)相关生物标志物和肝体比呈负相关。肝 DHA 和 EPA 水平与 elovl2 呈负相关,而 ARA 水平与 fadsd5 呈正相关。ABP 喂养鱼类肝 EPA/ARA 水平较低与与线粒体功能(fabp3a)、氧化应激(txna、prdx1b)和炎症(pgds、5loxa)相关的生物标志物表达增加有关。通过多重 PCR 分析肝生物标志物基因表达揭示了大西洋鲑在低水平海洋源性营养素摄入下的潜在生理影响和营养素-基因相互作用。