Effect of Temperature Lowering Rate on Physiological Response of Oplegnathus punctatus in Live Fish Chill Preservation
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摘要:
目的 研究冷驯化降温速率对斑石鲷生理应激的影响，缓解无水保活运输中斑石鲷（Oplegnathus punctatus）的应激反应。 方法 按照1、3、5 ℃·h−1的降温速率将斑石鲷从17.5～18.5 ℃降温至临界温度10 ℃致其休眠，并在该温度下进行无水保活，分别在0 h、2 h、4 h和6 h采样检测斑石鲷相关生理指标的变化情况。 结果 3 ℃·h−1降温处理组无水保活6 h后斑石鲷存活率最高，为76.7%。同时，随着无水保活时间的推移，各个降温处理组鱼的皮质醇（Cortisol）、谷草转氨酶（AST）、谷丙转氨酶（ALT）、丙二醛（MDA）、血糖（GLU）和乳酸（LAC）含量较降温前均显著升高（P＜0.05），而肾上腺素（EPI）、肝糖原（Liver glycogen）、肌糖原（Muscle glycogen）含量较降温前均显著降低（P＜0.05）；特别是当无水保活时间为6 h，处理组斑石鲷血清各指标较降温前差异最为显著（P＜0.05），其中，1 ℃·h−1降温处理组鱼的血清EPI、皮质醇以及肝脏ALT、肝脏肝糖原含量均显著高于3、5 ℃·h−1降温处理组（P＜0.05），表明1 ℃·h−1降温速率使斑石鲷产生了更加强烈的应激反应；5 ℃·h−1降温处理组试验鱼血清AST、MDA、LAC含量较1、3 ℃·h−1降温处理组显著升高（P＜0.05）；而3 ℃·h−1降温处理组鱼血清EPI、AST、ALT、MDA、LAC和肝脏肝糖原含量则显著低于1、5 ℃·h−1降温处理组（P＜0.05），表明3 ℃·h−1降温速率使斑石鲷产生的应激程度最小，肝损伤程度最低。 结论 以3 ℃·h−1降温速率冷驯化后无水保活6 h，斑石鲷产生的应激反应最低，建议将其冷驯化过程中的降温速率设为3 ℃·h−1。 Abstract:Objective Physiological response of Oplegnathus punctatus to the rate of temperature lowering was studied to optimize the live fish chill preservation technology for water-conservation transportation. Method Live and healthy O. punctatus were chilled at a rate of 1, 3, or 5 ℃·h−1 from 17.5–18.5 ℃ to the critical dormancy temperature of 10 ℃. Kept the fish out of water, in intervals of 0, 2, 4, and 6 h, sampling for the determination of various physiological indicators was conducted for a data analysis. Result The highest survival rate of O. punctatus in 6 h after the treatments was 76.7% with a chilling rate of 3 ℃·h−1. Under the chilling and low-temp preservation, the contents of cortisol, AST, ALT, MDA, GLU, and LAC in the fish of all treatment groups increased significantly (P＜0.05), while those of EPI, hepatic glycogen, and muscle glycogen significantly declined (P＜0.05). The serum indices of the fish chilled at all 3 rates for 6 h at 10 ℃ changed significantly (P＜0.05). The contents of EPI, cortisol, liver ALT, and hepatic glycogen were significantly higher under the 1 ℃·h−1 than the 3 ℃·h−1 or 5 ℃·h−1 treatment (P＜0.05) indicating a stronger stress response of the fish by the temperature lowering rate at 1 ℃·h−1 than the other two; while AST, MDA, and LAC were significantly higher under the 5 ℃·h−1 than the 1 ℃·h−1 or 3 ℃·h−1 treatment (P＜0.05) and EPI, AST, ALT, MDA, LAC, and hepatic glycogen significantly lower under the 3 ℃·h−1 than the 1 ℃·h−1 or 5 ℃·h−1 treatment (P＜0.05) showing the 3 ℃·h−1 rate of chilling minimized the effect of cold stress and lessen the liver injury on O. punctatus in the preservation process. Conclusion It appeared that a temperature lowering rate, such as 3 ℃·h−1, could be a tolerable threshold for the fish to make adequate acclimation to recover from physiological ill-effects with a high survival rate out of water at 10 ℃ for 6 h. -
表 1 不同降温速率下斑石鲷保活不同时间的存活率
Table 1. Survival rates of O. punctatus chilled at various temperature lowering rates out-of-water for different durations
降温速率
Temperature lowering rate/（℃·h−1）存活率 Survival rate/% 0 h 2 h 4 h 6 h 1 100.0 100.0 83.3 60.0 3 100.0 100.0 93.3 76.7 5 100.0 90.0 76.7 46.7 
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表 2 不同降温速率对斑石鲷无水保活中血清肾上腺素和皮质醇质量浓度的影响
Table 2. Effects of chilling rate on serum epinephrine and cortisol contents in survived O. punctatus under treatments
指标
Index组别
Group0 h 2 h 4 h 6 h 肾上腺素 Epinephrine/（ng·mL−1） 1 ℃·h−1 55.96±2.85 c 139.45±4.55 a 283.68±15.8 a 122.82±4.58 a 3 ℃·h−1 102.58±8.34 b 60.74±3.29 c 57.45±10.01 b 50.99±9.69 b 5 ℃·h−1 137.23±4.01 a 94.83±2.28 b 80.23±4.53 b 59.96±5.23 b 皮质醇 Cortisol/（ng·mL−1） 1 ℃·h−1 65.67±3.66 b 84.84±4.60 b 90.17±5.41 a 95.44±4.62 a 3 ℃·h−1 60.12±4.44 c 82.92±8.08 b 87.1±3.02 a 82.23±5.39 ab 5 ℃·h−1 92.1±5.17 a 95.59±5.83 a 73.79±6.08 b 65.89±4.85 b 注：同列数据后不同小写字母表示处理间差异显著（P＜0.05），相同小写字母者表示无显著差异（P＞0.05），下同。
Note：Different lowercase letters in the same sampling time indicated significant difference in the experimental groups(P＜0.05), the same lowercase letters indicated no significant difference in this index between the experimental groups(P＞0.05). The same below.
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表 3 不同降温速率对斑石鲷无水保活中血清谷草转氨酶、谷丙转氨酶和丙二醛活力的影响
Table 3. Effects of chilling rate on serum AST, ALT, and MDA activities in survived O. punctatus under treatments
指标
Index组别
Group0 h 2 h 4 h 6 h 谷草转氨酶 AST/（U·L−1） 1 ℃·h−1 151.24±5.32 b 202.05±7.23 b 243.53±7.14 a 279.45±5.82 b 3 ℃·h−1 142.82±4.16 b 174.23±6.45 c 225.19±5.63 b 229.37±4.35 c 5 ℃·h−1 174.03±3.68 a 239.83±7.56 a 252.43±9.17 a 304.98±9.16 a 谷丙转氨酶 ALT/（U·L−1） 1 ℃·h−1 30.91±2.14 a 45.36±3.21 a 54.28±4.33 a 70.04±4.65 a 3 ℃·h−1 26.82±3.23 b 33.72±3.82 c 42.04±2.37 b 47.59±3.54 c 5 ℃·h−1 24.89±2.65 b 39.89±4.12 b 51.83±1.29 a 62.27±5.23 b 丙二醛 MDA/（U·L−1） 1 ℃·h−1 2.19±0.18 b 2.86±0.10 b 4.74±0.23 c 7.34±0.35 b 3 ℃·h−1 1.68±0.15 c 2.37±0.04 c 5.23±0.23 b 6.47±0.17 c 5 ℃·h−1 3.70±0.17 a 4.65±0.15 a 6.52±0.35 a 7.87±0.31 a
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表 4 不同降温速率对斑石鲷无水保活中血糖、肝糖原、肌糖原和乳酸含量的影响
Table 4. Effects of chilling rate on GLU, hepatic glycogen, muscle glycogen, and LAC contents in survived O. punctatus under treatments
指标
Index组别
Group0 h 2 h 4 h 6 h 血糖 GLU/（mmol·L−1） 1 ℃·h−1 8.16±0.18 c 13.15±0.28 c 23.67±0.43 b 21.88±0.40 b 3 ℃·h−1 11.36±0.44 b 21.04±0.15 a 26.85±0.32 a 29.86±0.21 a 5 ℃·h−1 12.77±0.06 a 19.86±0.31 b 26.35±0.53 a 16.05±0.19 c 肝糖原 Hepatic Glycogen/（mg·g−1） 1 ℃·h−1 9.83±0.45 a 8.19±1.65 a 5.25±1.14 a 6.88±0.40 a 3 ℃·h−1 8.65±1.30 b 7.78±0.39 a 4.53±0.32 b 2.69±0.43 c 5 ℃·h−1 6.82±0.28 c 5.21±0.12 b 3.75±0.11 c 4.64±0.12 b 肌糖原 Muscle glycogen/（mg·g−1） 1 ℃·h−1 1.31±0.16 a 1.18±0.11 a 0.93±0.10 a 0.62±0.13 a 3 ℃·h−1 1.18±0.07 b 1.12±0.15 b 0.85±0.04 c 0.65±0.08 a 5 ℃·h−1 1.09±0.05 c 1.07±0.01 c 0.84±0.04 c 0.58±0.03 b 乳酸 LAC/（mmol·L−1） 1 ℃·h−1 6.14±0.03 c 5.72±0.14 c 8.97±0.12 b 10.28±0.16 b 3 ℃·h−1 6.61±0.18 b 5.86±0.21 b 8.62±0.23 b 9.97±0.22 c 5 ℃·h−1 6.93±0.13 a 6.32±0.16 a 9.93±0.27 a 11.31±0.28 a
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