鲫鱼和中国对虾的无水保活及冰温保鲜技术研究
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摘要
水产品由于其低脂肪、高蛋白的特点,已成为合理膳食营养结构中不可替代的组分,随着人们生活水平的提高,鲜活水产品越来越受到人们的青睐。近年来,开发高效的水产品保活保鲜技术研究已经成为人们关注的焦点。
本文探索了丁香酚麻醉对于鲫鱼无水保活的可行性,并测定了鲫鱼无水保活前后的品质变化和生化特性。丁香酚麻醉的鲫鱼装袋充入90%的氧气放在8℃、相对湿度95%-100%的环境下可无水保活38h,随后转入清水中可完全复活。保活前后持水性没有明显的变化,休眠下的鲫鱼TVB-N值、TBA值明显高于鲜活的鲫鱼。休眠过程中血清皮质醇浓度、GOT、AKP、ACP活性明显升高,但是复活后会立即下降,回到正常水平。实验证明,丁香酚麻醉无水保活鲫鱼的肌肉品质并不会受到影响,麻醉时产生的胁迫在复活后会立即消失。
鱼虾类冷血动物都有一个固定的生态冰温,在生态冰温区内,采用适当的降温,可使鱼虾处于假死的休眠状态,随后进行保活运输。鲫鱼在低温休眠状态下,血清中皮质醇浓度、溶菌酶活力和GOT活性显著上升,苏醒后恢复到正常状态。血清AKP活性在休眠状态下下降,苏醒后立即上升,而低温不会影响GPT和ACP的活性。结果表明低温无水保活下鲫鱼受到了应激胁迫,并做出了相应的胁迫响应,当放入清水中复活后,这种胁迫响应就会解除,恢复到正常状态。低温可能造成鲫鱼肝细胞的损伤和免疫能力的下降,但是,在放入清水中复活后,这种伤害会立即消失。经过低温休眠、复活后的鲫鱼与鲜活的鲫鱼状态一致。
鲫鱼经低温无水保活后,血清过氧化氢含量先下降再上升,超氧阴离子产生速率没有明显的变化。休眠状态下SOD、CAT和POD活性均明显地上升,复活后CAT活性下降,而SOD和POD活性没有变化。GSH-Px活性一直没有明显的差异。低温休眠下鲫鱼血清中SOD、CAT和GSH-Px的基因表达量均显著上升,放入清水中苏醒后,SOD和GSH-Px的基因表达量显著性降低,而CAT的表达量并没有明显差异。实验证明,鲫鱼在低温诱导休眠的过程中,体内的抗氧化防御系统被激活,这可能是其应对抗冷休克的主要机制,需要大量抗氧化酶表达来调整体内的动态平衡。
低温无水保活对鲫鱼的代谢水平也具有一定的影响。低温休眠下鲫鱼血液葡萄糖和乳酸浓度均有所升高,苏醒后恢复到正常状态。从新鲜到休眠再到复活后,LDH活性先降低后升高；而PK活性的变化趋势正好相反；HK活性一直呈现上升的趋势；PFK活性并没有显著性变化。结果表明低温降低鲫鱼的呼吸频率,对机体的供氧不足,体内组织转换为无氧代谢。在低温无水条件下保活38h,将鲫鱼放入清水中苏醒后,鲫鱼开始正常呼吸,各个指标逐渐恢复到新鲜水平,体内重新进行有氧代谢。
利用蛋白质组学技术筛选中国对虾在无水保活过程出现的差异蛋白并探讨其作用机制。通过双向电泳技术,每张凝胶图谱上大约有380-430个蛋白点被分离,其中,14个差异蛋白点被切取进行酶解和MALDI-TOF-MS分析。11个蛋白点被成功地鉴定,包括血蓝蛋白、血淋巴凝固蛋白、肌球蛋白轻链和基质钙结合蛋白。血蓝蛋白下调,血淋巴凝固蛋白和肌球蛋白轻链上调的共同作用使得对虾在低温下进入休眠状态并在无水条件下保持存活,而且,基质钙结合蛋白的上调使得对虾能够在温水中苏醒。本研究为对虾的低温无水保活提供了理论基础。
5%NaCl+5%CaCl2+2.5%山梨醇复配的冰点调节剂处理鲜活的中国对虾能有效地降低冰点,并抑制细菌的生长繁殖。冰点降低的对虾放在-2.0±0.2℃贮藏其肌肉的微观结构与新鲜的相比并没有什么变化,而对照组对虾肌肉内生成大量冰晶。以细菌总数、TVB-N值、TMA-N值和感官评定为指标,对虾冰点从-1.3℃降低到-2.5℃后放在-2.0±0.2℃贮藏,货架期可延长2天,同时具有较高的持水性、盐溶性蛋白含量和Ca2+-ATP激酶活性。实验证明,控制冰温贮藏具有很好的应用前景,即可保持对虾原有的鲜度,又可延长其货架期。
Aquatic product has become an irreplaceable componts in dietary nutrition and structure because of low fat and high protein. As the standard of living rise, fresh aquatic product is becoming more and more popular. In recent years, the research on preservation technology of aquatic product is highly attentioned.
The aim of this study was to explore the artificial hibernation of crucian carp for waterless preservation and to characterize the quality and biochemical properties during and after the hibernation. Anesthetized crucian carp using eugenol were stored at8℃with90%oxygen and95-100%relative humidity for38h, and then transferred to fresh water to recover. Liquid loss (LL) and cooking loss (CL) had no significant changes. The total volatile basic nitrogen (TVB-N) content and2-thiobarbituric acid (TBA) value in hibernated fish were significant higher than fresh and recovered groups. Serum cortisol, glutamic-oxaloacetic transaminase (GOT), alkaline phosphatase (AKP) and acid phosphatase (ACP) activities significantly increased during hibernation, while glutamic-pyruvic transaminase (GPT) had no significant change. Both ACP and AKP activities decreased upon the fish recovered, but only the ACP activity returned to normal. However, there were increased serum glucose concentration, GOT and GPT activities in recovered fish. The results indicated that muscle quality would not be influenced, and most of stress responses would disappear after hibernated fish recovered.
Low temperature can induce the fish or shrimp enter the hibernation state, and then subject waterless preservation. Serum cortisol concentration, lysozyme activity and GOT activity significantly increased in hibernated crucian carp compared to fresh sample, and then return to normal level after arouseing. Serum AKP activity offered upgrade firstly then descending tendency latter during and after hibernation There was no significant difference for ACP and GPT activities under low temperature. The results suggested that stress response under low temperature would be terminated when crucian carp was transferred to freahwater. The cell damage would disapper and immune function would be restored after recovered.
Serum H2O2content significantly decreased in the initiation of hibernation, while there was no significant difference compared to the fresh samples after arousal from hibernation. The O2-production rate had no significant changes among fresh, hibernated and aroused crucian carp. The total superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities of hibernated crucian carp significantly increased. CAT activity significantly decreased to normal, while SOD and POD activities had no significant changes after arousal compared to hibernated samples. However, glutathione peroxidase (GSH-Px) activity had no significant differences during and after the hibernation. The SOD、CAT and GSH-Px gene relative expression significantly increased in hibernated crucian carp serum, and then SOD、GSH-Px gene relative expression significantly decreased after recovered, but there was no significant difference for CAT gene relative expression. On the basis of these findings, the antioxidant defenses capacity was activated in response to the hibernation, which would be part of the adaptive mechanism for survival against the cold shock.
Waterless preservation under low temperature has certain effect on metabolism in crucian carp. Serum glucose and lactic acid concentration increased in hibernated crucian carp, and then return to normal level after recover. LDH activity declined firstly in hibernated sample and then rised in aroused crucian carp, while the changes of PK activity was just the opposite. HK activity had been rising during and after hibernation, however, there was no significant change for PFK activity. The results indicated that low temperature reduced the respiratory rate of crucian carp, which lead to hypoxia and anaerobic metabolism in tissue. The crucian carp began to breathe normally and aerobic metabolism in aroused crucian carp after waterless preservation for38h.
Comparative proteomic approach was used to identify the proteins that were changed in abundance during the waterless preservation of Chinese white shrimp (Fenneropenaeus chinensis). Approximately380-430protein spots were observed in the two dimensional protein electrophoresis (2-DE) gels. Among them,14altered proteins from hemolymph were excised and subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis.11protein spots were successfully identified, including hemocyanin, hemolymph clottable protein, myosin light chain and sarcoplasmic calcium-binding protein. It could be speculated that the combined action of decreased hemocyanin, increased hemolymph clottable protein and myosin light chain forced Chinese white shrimp into a temporary state of hibernation under cold temperature and could keep the hibernated shrimps alive during the waterless preservation. Moreover, increased expression of sarcoplasmic calcium-binding protein ensured that hibernated shrimps could be aroused at warm temperature. The present study provides a possible application of waterless preservation of Chinese white shrimp.
Incubation treatment with compound (5%NaCl,5%CaCl2and2.5%sorbitol) could effectively depress the freezing point and retard bacterial growth in the shrimps. There was little difference in microstructure between fresh and freezing-point depressed shrimps after storage at-2.0±0.2℃, while a large amount of ice crystals were observed in the control shrimps. Significant high WHC, SSP, and Ca2+-ATPase activity were obtained in freezing-point depressed shrimps. An additional2days of shelf life at-2.0±0.2℃was achieved in shrimps when the freezing-point was depressed from-1.3℃to-2.5℃considering TVC, TVB-N, TMA-N and sensory scores. Results suggested that controlled freezing-point storage may be a promising method of maintaining the freshness of Chinese white shrimp and extending shrimp post-mortem shelf-life.
本文探索了丁香酚麻醉对于鲫鱼无水保活的可行性,并测定了鲫鱼无水保活前后的品质变化和生化特性。丁香酚麻醉的鲫鱼装袋充入90%的氧气放在8℃、相对湿度95%-100%的环境下可无水保活38h,随后转入清水中可完全复活。保活前后持水性没有明显的变化,休眠下的鲫鱼TVB-N值、TBA值明显高于鲜活的鲫鱼。休眠过程中血清皮质醇浓度、GOT、AKP、ACP活性明显升高,但是复活后会立即下降,回到正常水平。实验证明,丁香酚麻醉无水保活鲫鱼的肌肉品质并不会受到影响,麻醉时产生的胁迫在复活后会立即消失。
鱼虾类冷血动物都有一个固定的生态冰温,在生态冰温区内,采用适当的降温,可使鱼虾处于假死的休眠状态,随后进行保活运输。鲫鱼在低温休眠状态下,血清中皮质醇浓度、溶菌酶活力和GOT活性显著上升,苏醒后恢复到正常状态。血清AKP活性在休眠状态下下降,苏醒后立即上升,而低温不会影响GPT和ACP的活性。结果表明低温无水保活下鲫鱼受到了应激胁迫,并做出了相应的胁迫响应,当放入清水中复活后,这种胁迫响应就会解除,恢复到正常状态。低温可能造成鲫鱼肝细胞的损伤和免疫能力的下降,但是,在放入清水中复活后,这种伤害会立即消失。经过低温休眠、复活后的鲫鱼与鲜活的鲫鱼状态一致。
鲫鱼经低温无水保活后,血清过氧化氢含量先下降再上升,超氧阴离子产生速率没有明显的变化。休眠状态下SOD、CAT和POD活性均明显地上升,复活后CAT活性下降,而SOD和POD活性没有变化。GSH-Px活性一直没有明显的差异。低温休眠下鲫鱼血清中SOD、CAT和GSH-Px的基因表达量均显著上升,放入清水中苏醒后,SOD和GSH-Px的基因表达量显著性降低,而CAT的表达量并没有明显差异。实验证明,鲫鱼在低温诱导休眠的过程中,体内的抗氧化防御系统被激活,这可能是其应对抗冷休克的主要机制,需要大量抗氧化酶表达来调整体内的动态平衡。
低温无水保活对鲫鱼的代谢水平也具有一定的影响。低温休眠下鲫鱼血液葡萄糖和乳酸浓度均有所升高,苏醒后恢复到正常状态。从新鲜到休眠再到复活后,LDH活性先降低后升高；而PK活性的变化趋势正好相反；HK活性一直呈现上升的趋势；PFK活性并没有显著性变化。结果表明低温降低鲫鱼的呼吸频率,对机体的供氧不足,体内组织转换为无氧代谢。在低温无水条件下保活38h,将鲫鱼放入清水中苏醒后,鲫鱼开始正常呼吸,各个指标逐渐恢复到新鲜水平,体内重新进行有氧代谢。
利用蛋白质组学技术筛选中国对虾在无水保活过程出现的差异蛋白并探讨其作用机制。通过双向电泳技术,每张凝胶图谱上大约有380-430个蛋白点被分离,其中,14个差异蛋白点被切取进行酶解和MALDI-TOF-MS分析。11个蛋白点被成功地鉴定,包括血蓝蛋白、血淋巴凝固蛋白、肌球蛋白轻链和基质钙结合蛋白。血蓝蛋白下调,血淋巴凝固蛋白和肌球蛋白轻链上调的共同作用使得对虾在低温下进入休眠状态并在无水条件下保持存活,而且,基质钙结合蛋白的上调使得对虾能够在温水中苏醒。本研究为对虾的低温无水保活提供了理论基础。
5%NaCl+5%CaCl2+2.5%山梨醇复配的冰点调节剂处理鲜活的中国对虾能有效地降低冰点,并抑制细菌的生长繁殖。冰点降低的对虾放在-2.0±0.2℃贮藏其肌肉的微观结构与新鲜的相比并没有什么变化,而对照组对虾肌肉内生成大量冰晶。以细菌总数、TVB-N值、TMA-N值和感官评定为指标,对虾冰点从-1.3℃降低到-2.5℃后放在-2.0±0.2℃贮藏,货架期可延长2天,同时具有较高的持水性、盐溶性蛋白含量和Ca2+-ATP激酶活性。实验证明,控制冰温贮藏具有很好的应用前景,即可保持对虾原有的鲜度,又可延长其货架期。
Aquatic product has become an irreplaceable componts in dietary nutrition and structure because of low fat and high protein. As the standard of living rise, fresh aquatic product is becoming more and more popular. In recent years, the research on preservation technology of aquatic product is highly attentioned.
The aim of this study was to explore the artificial hibernation of crucian carp for waterless preservation and to characterize the quality and biochemical properties during and after the hibernation. Anesthetized crucian carp using eugenol were stored at8℃with90%oxygen and95-100%relative humidity for38h, and then transferred to fresh water to recover. Liquid loss (LL) and cooking loss (CL) had no significant changes. The total volatile basic nitrogen (TVB-N) content and2-thiobarbituric acid (TBA) value in hibernated fish were significant higher than fresh and recovered groups. Serum cortisol, glutamic-oxaloacetic transaminase (GOT), alkaline phosphatase (AKP) and acid phosphatase (ACP) activities significantly increased during hibernation, while glutamic-pyruvic transaminase (GPT) had no significant change. Both ACP and AKP activities decreased upon the fish recovered, but only the ACP activity returned to normal. However, there were increased serum glucose concentration, GOT and GPT activities in recovered fish. The results indicated that muscle quality would not be influenced, and most of stress responses would disappear after hibernated fish recovered.
Low temperature can induce the fish or shrimp enter the hibernation state, and then subject waterless preservation. Serum cortisol concentration, lysozyme activity and GOT activity significantly increased in hibernated crucian carp compared to fresh sample, and then return to normal level after arouseing. Serum AKP activity offered upgrade firstly then descending tendency latter during and after hibernation There was no significant difference for ACP and GPT activities under low temperature. The results suggested that stress response under low temperature would be terminated when crucian carp was transferred to freahwater. The cell damage would disapper and immune function would be restored after recovered.
Serum H2O2content significantly decreased in the initiation of hibernation, while there was no significant difference compared to the fresh samples after arousal from hibernation. The O2-production rate had no significant changes among fresh, hibernated and aroused crucian carp. The total superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities of hibernated crucian carp significantly increased. CAT activity significantly decreased to normal, while SOD and POD activities had no significant changes after arousal compared to hibernated samples. However, glutathione peroxidase (GSH-Px) activity had no significant differences during and after the hibernation. The SOD、CAT and GSH-Px gene relative expression significantly increased in hibernated crucian carp serum, and then SOD、GSH-Px gene relative expression significantly decreased after recovered, but there was no significant difference for CAT gene relative expression. On the basis of these findings, the antioxidant defenses capacity was activated in response to the hibernation, which would be part of the adaptive mechanism for survival against the cold shock.
Waterless preservation under low temperature has certain effect on metabolism in crucian carp. Serum glucose and lactic acid concentration increased in hibernated crucian carp, and then return to normal level after recover. LDH activity declined firstly in hibernated sample and then rised in aroused crucian carp, while the changes of PK activity was just the opposite. HK activity had been rising during and after hibernation, however, there was no significant change for PFK activity. The results indicated that low temperature reduced the respiratory rate of crucian carp, which lead to hypoxia and anaerobic metabolism in tissue. The crucian carp began to breathe normally and aerobic metabolism in aroused crucian carp after waterless preservation for38h.
Comparative proteomic approach was used to identify the proteins that were changed in abundance during the waterless preservation of Chinese white shrimp (Fenneropenaeus chinensis). Approximately380-430protein spots were observed in the two dimensional protein electrophoresis (2-DE) gels. Among them,14altered proteins from hemolymph were excised and subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis.11protein spots were successfully identified, including hemocyanin, hemolymph clottable protein, myosin light chain and sarcoplasmic calcium-binding protein. It could be speculated that the combined action of decreased hemocyanin, increased hemolymph clottable protein and myosin light chain forced Chinese white shrimp into a temporary state of hibernation under cold temperature and could keep the hibernated shrimps alive during the waterless preservation. Moreover, increased expression of sarcoplasmic calcium-binding protein ensured that hibernated shrimps could be aroused at warm temperature. The present study provides a possible application of waterless preservation of Chinese white shrimp.
Incubation treatment with compound (5%NaCl,5%CaCl2and2.5%sorbitol) could effectively depress the freezing point and retard bacterial growth in the shrimps. There was little difference in microstructure between fresh and freezing-point depressed shrimps after storage at-2.0±0.2℃, while a large amount of ice crystals were observed in the control shrimps. Significant high WHC, SSP, and Ca2+-ATPase activity were obtained in freezing-point depressed shrimps. An additional2days of shelf life at-2.0±0.2℃was achieved in shrimps when the freezing-point was depressed from-1.3℃to-2.5℃considering TVC, TVB-N, TMA-N and sensory scores. Results suggested that controlled freezing-point storage may be a promising method of maintaining the freshness of Chinese white shrimp and extending shrimp post-mortem shelf-life.
引文
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