Effects of nanometer selenium on the immune protection and antioxidant ability of Eriocheir sinensis under hypoxia stress
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摘要: 为探究纳米硒 (nano-Se) 对低氧胁迫下中华绒螯蟹 (Eriocheir sinensis) 免疫保护和抗氧化能力的影响，该研究在基础饲料中添加不同水平 (0、0.1、0.2、0.4、0.8和1.6 mg·kg−1) 的纳米硒饲喂中华绒螯蟹90 d。饲喂实验结束后，进行低氧胁迫实验并注射嗜水气单胞菌 (Aeromonas hydrophila)。实验结果表明：1) 低氧胁迫24 h和低氧胁迫下感染嗜水气单胞菌24 h的蟹死亡率分别可达62.45%和100%，低氧胁迫12 h使血淋巴中血蓝蛋白含量、血细胞数量、组织中超氧化物歧化酶 (SOD)、过氧化氢酶 (CAT)、谷胱甘肽过氧化物酶 (GSH-Px) 活性显著升高 (P<0.05)，胁迫至24 h有下降趋势；乳酸 (LD) 和丙二醛 (MDA) 含量在低氧胁迫12~24 h持续上升。2) 饲料中添加适量 (0.1~0.4 mg·kg−1) 纳米硒可显著缓解低氧胁迫下蟹死亡率和低氧胁迫下嗜水气单胞菌的致蟹死亡率 (P<0.05)，显著提高低氧胁迫下血蓝蛋白含量和血细胞数量以及抗氧化酶 (SOD、CAT、GSH-Px) 活性，降低LD和MDA含量 (P<0.05)；添加0.8~1.6 mg·kg−1水平纳米硒加剧了低氧胁迫损伤。结果表明饲料中添加适量纳米硒可改善低氧胁迫下中华绒螯蟹的免疫功能和抗氧化能力，且添加水平以0.2 mg·kg−1为宜。Abstract: In order to investigate the effects of nanometer selenium (nano-Se) on immune protection and antioxidant capacity of Eriocheir sinensis under hypoxia stress, we fed E. sinensis with different doses of nano-Se (0, 0.1, 0.2, 0.4, 0.8 and 1.6 mg·kg−1) in basic diets for 90 d. After the feeding, we conducted a hypoxia stress test and injected Aeromonas hydrophila under hypoxia stress. The results show that: 1) The mortality of E. sinensis under hypoxia stress and E. sinensis infected with A. hydrophila under hypoxia stress reached 62.45% and 100%, respectively. The levels of hemocyanin and the hemocyte count in crab hemolymph, and the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in crab tissues increased significantly under hypoxia stress for 12 h (P<0.05), and there was a downward trend under stress to 24 h. The contents of lactic acid (LD) and malondialdehyde (MDA) continued to rise from 12 to 24 h under hypoxia stress. 2) Appropriate addition amount (0.1−0.4 mg·kg−1) of nano-Se reduced the mortality of E. sinensis significantly under hypoxia stress and the lethality of A. hydrophila under hypoxia stress (P<0.05), increasing the levels of hemocyanin and the hemocyte count and the activities of antioxidant enzymes (SOD, CAT, GSH-Px) under hypoxia stress significantly, but decreasing the contents of LD and MDA (P<0.05). The addition of 0.8−1.6 mg·kg−1 nano-Se had aggravated hypoxia stress injury. These results indicate that appropriate addition of nano-Se to the diets can improve the decrease of immune response and antioxidant ability of E. sinensis under hypoxia stress, and the optimal dose of nano-Se in basal diets is 0.2 mg·kg−1.
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Key words:
- Nanometer selenium /
- Hypoxia stress /
- Eriocheir sinensis /
- Immunity /
- Antioxidant
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图 1 纳米硒对低氧胁迫下中华绒螯蟹血淋巴耐低氧指标的影响
不同英文字母表示相同低氧胁迫时间下显著差异 (P<0.05)；*. 与对照组差异显著 (P<0.05)；后图同此
Figure 1. Effects of nano-Se on hemolymph hypoxia tolerance indexes of E. sinensis under hypoxia stress
Different lowercase letters indicate significant difference at the same hypoxia stress time (P<0.05); *. Significant difference compared with the control group (P<0.05); the same case in the following figures.
图 2 纳米硒对低氧胁迫下中华绒螯蟹血淋巴血细胞数量的影响
Figure 2. Effects of nano-Se on hemolymph hemocyte count of E. sinensis under hypoxia stress
图 3 纳米硒对低氧胁迫下中华绒螯蟹不同组织中超氧化物歧化酶活性的影响
Figure 3. Effects of nano-Se on SOD activity in different tissues of E. sinensis under hypoxia stress
图 4 纳米硒对低氧胁迫下中华绒螯蟹不同组织中过氧化氢酶活性的影响
Figure 4. Effects of nano-Se on CAT activity in different tissues of E. sinensis under hypoxia stress
图 5 纳米硒对低氧胁迫下中华绒螯蟹不同组织中谷胱甘肽过氧化物酶活性的影响
Figure 5. Effects of nano-Se on GSH-Px activity in different tissues of E. sinensis under hypoxia stress
图 6 纳米硒对低氧胁迫下中华绒螯蟹不同组织中丙二醛含量的影响
Figure 6. Effects of nano-Se on MDA content in different tissues of E. sinensis under hypoxia stress
表 1 基础饲料配方组成
Table 1. Ingredients of composition of basal diets
原料 Ingredient 含量 Content/% 鱼粉 Fish meal 17 棉粕 Cotton seed meal 17 菜粕 Rape seed meal 16 次粉 Wheat flour 10.5 豆粕 Soybean meal 10 玉米 Corn 9 米糠 Rice bran 5 粘合剂 Adhesive 1 血粉 Blood meal 3 虾壳粉 Shrimp shell meal 3 豆油+菜油 (1:1) Soybean oil+ Rapeseed oil 2 磷酸二氢钙 Ca(H2PO4)2 1.5 沸石粉 Zeolite powder 2 河蟹饲料添加剂 Crab feed additive 1 蟹用多维a Crab vit premix 1 蟹用多矿b Crab min premix 1 注：a. 每100 g蟹用多维预混料中含：2.0 g维生素E、3.0 g维生素C、0.6 g维生素A、0.08 g维生素D3、0.07 g维生素B1、0.14 g维生素B2、0.28 g维生素B3、0.01 g维生素B5、0.08 g维生素B6、0.05 g维生素B7、0.02 g维生素B11、0.04 g维生素H、0.3 g烟酸、0.05 g叶酸、0.5 g氯化胆碱、0.25 g泛酸钙、0.05 g生物素、0.7 g肌醇；b. 每100 g蟹用多矿预混料中含：3.5 g磷酸二氢钠、6.0 g磷酸二氢钾、3.5 g碳酸钙、0.6 g氯化钾、3.2 g七水合硫酸镁、0.55 g六水合氯化铝、0.157 g七水合硫酸锌、0.019 g柠檬酸铁、0.043 g四水合硫酸锰、0.016 g碘化钾、0.014 g氯化铜、0.055 g六水合氯化钴、5.15 g乳酸钙。 Note: a. Per 100g of crab multidimensional premix contains: 2.0 g VE, 3.0 g VC, 0.6 g VA, 0.08 g VD3, 0.07 g VB1, 0.14 g VB2, 0.28 g VB3, 0.01 g VB5, 0.08 g VB6, 0.05 g VB7, 0.02 g VB11, 0.04 g VH, 0.3 g niacin, 0.05 g folic acid, 0.5 g choline chloride, 0.25 g calcium pantothenate, 0.05 g biotin, 0.7 g inositol; b. The crab min premix provided following for per 100 g: 3.5 g NaH2PO4, 6.0 g KH2PO4, 3.5 g CaCO3, 0.6 g KCl, 3.2 g MgSO4·7H2O, 0.55 g AlCl3·6H2O, 0.157 g ZnSO4·7H2O, 0.019 g FeC6H5O7, 0.043 g MnSO4·4H2O, 0.016 g KI, 0.014 g CuCl2, 0.055 g CoCl2·6H2O, 5.15 g C6H10CaO6. 
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表 2 纳米硒对低氧胁迫下中华绒螯蟹死亡率的影响 (`X±SE, N=10)
Table 2. Effects of nano-Se on mortality rate of E. sinensis under hypoxia stress
纳米硒添加水平
Level of nano-Se/(mg·kg−1)死亡率
Mortality rate/%免疫保护率
Immune protection rate/%0 h 12 h 24 h 12 h 24 h 0 0 12.66±3.33c 62.45±8.57c* — — 0.1 0 6.25±1.50b 44.39±4.88b 50.63 28.92 0.2 0 2.50±0.30a 25.62±3.33a 80.25 58.98 0.4 0 12.34±2.05c 31.55±3.29b 2.53 49.48 0.8 0 19.36±2.67d 62.66±6.32c −52.92 −0.34 1.6 0 20.25±5.55d 87.73±10.34d −59.95 −40.48 注：同列不同上标字母表示差异显著 (P<0.05)，*. 与对照组 (0 mg·kg−1纳米硒低氧胁迫0 h组) 差异显著 (P<0.05)；—. 无数据；后表同此。 Note: Different superscript letters within the same column indicate significant difference (P<0.05); *. Significant difference compared with the control group (0 mg·kg−1 nano-Se group under hypoxia stress for 0 h) (P<0.05); —. No data. The same case in the following tables.
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表 3 纳米硒对低氧胁迫下嗜水气单胞菌致死率的影响 (`X±SE, N=10)
Table 3. Effects of nano-Se on lethality of A. hydrophila under hypoxia stress
纳米硒添加水平
Level of nano-Se/(mg·kg−1)嗜水气单胞菌致死率
Lethality of A. hydrophila/%免疫保护率
Immune protection rate/%0 h 12 h 24 h 12 h 24 h 0 0 50.36±6.72c* 100±0c* — — 0.1 0 25.22±3.51b 87.47±5.95b 49.92 12.53 0.2 0 12.48±2.39a 56.82±7.31a 75.22 43.18 0.4 0 25.78±3.64b 81.43±2.5b 48.81 18.57 0.8 0 50.33±4.52c 100±0c 0.06 0 1.6 0 75.29±6.15d 100±0c −49.5 0
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