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养殖温度对生长激素转基因三倍体雌性大西洋鲑(Salmo salar)固有抗病毒免疫反应的影响。

Impact of rearing temperature on the innate antiviral immune response of growth hormone transgenic female triploid Atlantic salmon (Salmo salar).

机构信息

AquaBounty Canada, 718 Route 310, Fortune, PE, C0A 2B0, Canada; Memorial University, Fisheries and Marine Institute, 155 Ridge Road, St. John's, NL, A1C 5R3, Canada; Memorial University, Department of Ocean Sciences, 0 Marine Lab Road, St. John's, NL, A1C 5S7, Canada.

AquaBounty Canada, 718 Route 310, Fortune, PE, C0A 2B0, Canada; Hoplite Laboratory, Department of Pathology and Microbiology, Atlantic Veterinary College, University of Prince Edward Island, 550 University Ave., Charlottetown, PE, C1A 4P3, Canada.

出版信息

Fish Shellfish Immunol. 2020 Feb;97:656-668. doi: 10.1016/j.fsi.2019.12.081. Epub 2019 Dec 28.

Abstract

AquAdvantage Salmon (growth hormone transgenic female triploid Atlantic salmon) are a faster-growing alternative to conventional farmed diploid Atlantic salmon. To investigate optimal rearing conditions for their commercial production, a laboratory study was conducted in a freshwater recirculating aquaculture system (RAS) to examine the effect of rearing temperature (10.5 °C, 13.5 °C, 16.5 °C) on their antiviral immune and stress responses. When each temperature treatment group reached an average weight of 800 g, a subset of fish were intraperitoneally injected with either polyriboinosinic polyribocytidylic acid (pIC, a viral mimic) or an equal volume of sterile phosphate-buffered saline (PBS). Blood and head kidney samples were collected before injection and 6, 24 and 48 h post-injection (hpi). Transcript abundance of 7 antiviral biomarker genes (tlr3, lgp2, stat1b, isg15a, rsad2, mxb, ifng) was measured by real-time quantitative polymerase chain reaction (qPCR) on head kidney RNA samples. Plasma cortisol levels from blood samples collected pre-injection and from pIC and PBS groups at 24 hpi were quantified by ELISA. While rearing temperature and treatment did not significantly affect circulating cortisol, all genes tested were significantly upregulated by pIC at all three temperatures (except for tlr3, which was only upregulated in the 10.5 °C treatment). Target gene activation was generally observed at 24 hpi, with most transcript levels decreasing by 48 hpi in pIC-injected fish. Although a high amount of biological variability in response to pIC was evident across all treatments, rearing temperature significantly influenced transcript abundance and/or fold-changes comparing time- and temperature-matched pIC- and PBS-injected fish for several genes (tlr3, lgp2, stat1b, isg15a, rsad2 and ifng) at 24 hpi. As an example, significantly higher fold-changes of rsad2, isg15a and ifng were found in fish reared at 10.5 °C when compared to 16.5 °C. Multivariate analysis confirmed that rearing temperature modulated antiviral immune response. The present experiment provides novel insight into the relationship between rearing temperature and innate antiviral immune response in AquAdvantage Salmon.

摘要

AquAdvantage 三文鱼(生长激素转基因雌性三倍体大西洋三文鱼)是传统养殖的二倍体大西洋三文鱼的一种生长速度更快的替代品。为了研究其商业生产的最佳养殖条件,本研究在淡水循环水产养殖系统(RAS)中进行了一项实验室研究,以检验养殖温度(10.5°C、13.5°C、16.5°C)对其抗病毒免疫和应激反应的影响。当每个温度处理组的鱼体重达到 800g 左右时,将一部分鱼用多聚肌苷酸多聚胞苷酸(pIC,病毒模拟物)或等量无菌磷酸盐缓冲盐水(PBS)进行腹腔内注射。在注射前和注射后 6、24 和 48 小时(hpi)收集血液和头肾样本。通过实时定量聚合酶链反应(qPCR)对头肾 RNA 样本中 7 种抗病毒生物标志物基因(tlr3、lgp2、stat1b、isg15a、rsad2、mxb、ifng)的转录丰度进行测量。通过 ELISA 定量测定注射前和注射后 24 小时 pIC 和 PBS 组的血液样本中的血浆皮质醇水平。虽然养殖温度和处理对循环皮质醇没有显著影响,但所有测试基因在所有三个温度下均被 pIC 显著上调(10.5°C 处理组中的 tlr3 除外,该基因仅被上调)。在 24 hpi 时通常观察到靶基因的激活,在 pIC 注射鱼中,大多数基因的转录水平在 48 hpi 时下降。尽管所有处理中都明显存在对 pIC 反应的大量生物学变异性,但在 24 hpi 时,与时间和温度匹配的 pIC 和 PBS 注射鱼相比,养殖温度显著影响了几种基因(tlr3、lgp2、stat1b、isg15a、rsad2 和 ifng)的转录丰度和/或倍数变化。例如,在 10.5°C 养殖的鱼中,rsad2、isg15a 和 ifng 的倍数变化明显高于 16.5°C 养殖的鱼。多元分析证实,养殖温度调节了抗病毒免疫反应。本实验为 AquAdvantage 三文鱼养殖温度与先天抗病毒免疫反应之间的关系提供了新的见解。

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