操作及低温胁迫对四指马鲅幼鱼组织结构和抗氧化系统的影响

发布时间：2018-06-02 04:28

本文选题：四指马鲅 + 操作胁迫　；参考：《上海海洋大学》2017年硕士论文

【摘要】：为探究操作、运输胁迫对四指马鲅(Eleutheronema tetradactylum)幼鱼组织结构及抗氧化系统的影响,本研究通过组织切片以及相关指标的测定技术,对试验结果进行了分析对比,表明不同的胁迫方式对四指马鲅幼鱼造成不同程度的损伤,同时也反应出四指马鲅幼鱼对应激胁迫的应答规律,其主要研究结果如下:1急性操作胁迫对四指马鲅幼鱼肝脏、鳃、肌肉组织结构和氧化应激的影响为探究急性操作胁迫对四指马鲅幼鱼组织结构以及氧化应激的影响。该研究在离水操作胁迫后不同时间取样,来观察肝脏显微结构的变化,以及检测相关抗氧化酶活性。结果显示:随着离水胁迫时间的延长,肝脏组织损伤程度出现一个先升后降的变化趋势,胁迫24 h肝脏基本结构与对照组相似;胁迫2 h后鳃小片、扁平上皮细胞以及线粒体丰富细胞都出现不同程度的损伤;肌肉从肌纤维变性、肌纤维束之间的间隙增宽,空泡化以至于肌肉组织整体失去固有形态,并且呈逐渐分解的趋势。超氧化物歧化酶(SOD)和过氧化氢酶(CAT)在肝脏中,有相同的趋势,最后其值显著(P0.05)低于对照组;肌肉中SOD和还原型谷胱甘肽(GSH)有相同的变化趋势,胁迫后的2 h显著下降(P0.05),但12 h则反升至最高值。肝脏中CAT和总抗氧化能力(T-AOC)处理后2 h其水平无显著变化(P0.05),之后升高直到12 h达到峰值。肝脏中丙二醛(MDA)含量在胁迫2~12h出现先升高后下降的变化趋势,12 h以后又有所上升;肌肉中MDA含量在胁迫后的2 h无显著差异(P0.05),直到6 h达到峰值。肝脏中GSH含量在胁迫2h没有显著变化(P0.05),最终其含量仍显著(P0.05)低于对照组;肌肉T-AOC在胁迫2h显著上升(P0.05),之后下降,试验结束时显著低于(P0.05)对照组。肌肉中SOD和CAT在处理24 h后活性显著低于处理前,其他酶活性变化不显著。Na+-K+-ATP酶活性在处理后2 h显著升高(P0.05)12 h达到峰值,24 h恢复到处理前水平。综合以上结果得出:离水操作胁迫确实对四指马鲅幼鱼肝脏组织结构以及抗氧化酶产生一定的影响,但随着胁迫时间的延长,肝脏组织结构以及抗氧化能力都有一定的恢复趋势,而鳃和肌肉则随时间持续损伤呈现加重趋势。2运输胁迫对四指马鲅幼鱼组织结构、抗氧化系统的影响以及抗应激剂混合物的作用为探究运输胁迫对四指马鲅幼鱼组织结构、抗氧化系统的影响以及抗应激剂的作用。该研究设置了对照组(未运输)和实验组,其中实验组包括:空白运输组和抗应激剂组即运输过程中添加抗应激剂混合物(维生素C:谷氨酰胺=1:1);实验组经过运输6h后分别取样。结果显示:添加抗应激剂组肝脏部分区域细胞排列疏松,肝板结构不清晰;空白运输组肝脏组织整体失去固有形态,肝板结构消失不见。添加抗应激剂组的鳃丝排列混乱,线粒体丰富细胞体积变大;空白运输组鳃小片整体肿胀,部分鳃小片因红细胞过多而涨破,线粒体丰富细胞体积变大且数量增多。添加抗应激剂组的脾脏白髓区域相对增加,黑色素-巨噬细胞中心数量以及大小都有所增加;空白运输组脾脏组织红髓区域占据大部分,白髓区域相对较少,巨噬细胞中心减小。肝脏和肌肉中SOD活性值均显著(P0.05)的表现为对照组混合组空白组,且肝脏中其活性值远高于肌肉中;肝脏中CAT活性值显著(P0.05)的表现为空白组对照组混合组,而肌肉中则为混合组对照组空白组;肝脏MDA含量显著(P0.05)的表现为空白组混合组对照组,肌肉中其值表明为对照组与混合组之间无显著(P0.05)差异,而两组均显著(P0.05)低于空白组,而肌肉中其含量值远高于肝脏中;肌肉中GSH含量显著(P0.05)的表现为空白组混合组对照组,肝脏中其值表明为对照组与混合组之间无显著(P0.05)差异,而两组均显著(P0.05)低于空白组;肝脏T-AOC值表现为显著(P0.05)的空白组混合组对照组,而肌肉中则混合组与空白组之间无显著(P0.05)差异,且显著(P0.05)低于对照组;对照组鳃钠钾ATP酶活性值略高于混合组但不显著(P0.05),混合组显著(P0.05)高于空白组。该研究结果为四指马鲅鱼苗的运输,提高运输成活率提供参考资料。3运输胁迫对四指马鲅幼鱼抗氧化系统的影响以及抗应激剂的作用为探究运输胁迫对四指马鲅幼鱼肝脏、肌肉抗氧化系统的影响以及抗应激剂的生理作用。该研究设置了对照组、空白组、维生素C组以及谷氨酰胺组,在运输试验进行的2h、6h以及9h分别采集肝脏以及肌肉样品进行相关指标的测定。测定项目包括:SOD,CAT,MDA,GSH,T-AOC。结果显示:运输胁迫使空白组肝脏和肌肉中SOD活性迅速升高,而抗应激剂的存在使其最终活性低于对照组;运输胁迫使空白组肝脏和肌肉中CAT活性显著(P0.05)升高,虽然之后有所下将,但最后其活性值均显著(P0.05)高于对照组,而维生素C组和谷氨酰胺组最终值与对照组无显著差异(P0.05)或略高于对照组;运输胁迫使空白组肝脏和肌肉MDA含量均显著(P0.05)升高,而抗应激剂明显缓解了这一结果;GSH含量在肝脏与肌肉中出现了不同的变化趋势,可能与二者所承担的生理功能不同有关;运输胁迫后,空白组肝脏与肌肉T-AOC值均显著(P0.05)高于对照组,而维生素C组和谷氨酰胺组最终则略高于或等于对照组。本研究分析了运输胁迫下四指马鲅幼鱼抗氧化系统变化规律以及抗应激的作用效果,为其人工养殖的推广提供参考依据。4急性低温对四指马鲅幼鱼组织结构、抗氧化系统以及鳃钠钾ATP酶活性的影响为探究急性低温胁迫对四指马鲅幼鱼组织结构、抗氧化系统以及鳃钠钾ATP酶活性的影响。该研究设置了常温(27±0.5℃)组、20℃组以及15℃组,三个温度梯度进行试验,以常温组作为对照,并在试验进行的2h、6h以及12h分别采集肝脏、肌肉、脾以及鳃样品进行观察和测定。结果显示:随着处理时间的延长,20℃组的肝脏细胞空泡结构呈增多,细胞核偏离,染色变浅趋势;15℃组肝脏细胞排列混乱,肝板结构不清晰,直到最后细胞核溶解,肝脏整体失去固有形态。随着低温处理时间的延长20℃组肌纤维出现轻微的弯曲现象到大部分肌纤维弯曲,肌纤维之间与内部均出现较大间隙;15℃组肌纤维间隙增大,部分断裂直到肌纤维之间与内部均严重开裂,部分肌纤维溶解并暴露出细胞核。随着低温处理时间的延长20℃组主要表现为线粒体丰富细胞数目增多,鳃小片末端轻微的膨大到鳃小片整体水肿且严重弯曲,血管以及血窦内出现大量的红细胞,部分鳃小片因红细胞过多而涨破;15℃组鳃小片表皮出现轻微的脱落,少部分鳃小片胀大到其吸水涨破,鳃小片基本形态不可见。SOD在肝脏和肌肉中,两个温度下均出现先升高后降低的变化趋势,而且最终都显著(P0.05)低于对照组;两个温度下CAT在肝脏中出现均出现先升高后降低的趋势,而在肌肉中两个温度下变化趋势不同,但最终其活性值均显著(P0.05)低于对照组;GSH在肝脏和肌肉中,两个温度下均出现下降趋势,不同的是20℃的肌肉中出现一直显著下降趋势,而在肝脏以及15℃的肌肉中则是先下降后保持在一定值得趋势;MDA在肝脏的两个温度下以及20℃条件下的肌肉中均出现先升高后降低的趋势,而在15℃的肌肉中先升高保持一段时间后又继续升高的趋势;T-AOC在肝脏的两个温度下以及15℃条件下的肌肉中均出现一直下降趋势,而在20℃下的肌肉中则是下升高后下降的趋势。鳃Na+-K+-ATP酶活性在两个温度下均出现先下降后上升的趋势,不同的是20℃条件下处于一直上升趋势,而15℃条件下则上升到一定值保持不变。该研究结果为四指马鲅鱼苗的越冬管理,提高越冬成活率提供参考资料。
[Abstract]:In order to explore the effect of transportation stress on the tissue structure and antioxidant system of four fingerling mackerel (Eleutheronema tetradactylum) young fish, the results of this study were analyzed and compared by tissue section and related indexes, which showed that different stress modes caused different degrees of damage to the four fingerlings of Spanish mackerel. The response of four fingers to young mackerel young fish was also observed. The main results were as follows: 1 the effects of acute operation stress on the liver, gill, muscle structure and oxidative stress of four fingers mackerel were affected by the effects of acute operation stress on the tissue structure and oxidative stress of four finger mackerel young fish. The changes in the liver microstructure and the activity of related antioxidant enzymes were observed at different time after pressure. The results showed that the degree of liver tissue damage appeared to rise first and then descend with the prolongation of the time of water stress, and the basic structure of 24 h of the liver was similar to that in the control group, and the lamellae and flat epithelial cells after 2 h stress were stressed. And the mitochondria rich cells have different degrees of damage; muscle from muscle fiber denaturation, the gap between the muscle fiber bundles widened, vacuolization so that the whole muscle tissue lost the inherent morphology, and showed a gradual trend of decomposition. Superoxide dismutase (SOD) and hydrogen peroxide (CAT) in the liver, the same trend, and finally its value (P0.05) was lower than the control group; SOD and glutathione (GSH) in muscle had the same trend of change, and the 2 h after stress decreased significantly (P0.05), but 12 h rose to the highest value. There was no significant change in the level of CAT and total antioxidant capacity (T-AOC) in the liver (P0.05), and then reached the peak until 12 h. The liver malondialdehyde (MDA) After 12 h, the content of the content increased after 12 h, and the content of MDA in the muscle was not significantly different (P0.05) after stress (P0.05), until 6 h reached its peak. The content of GSH in the liver was not significantly changed (P0.05), and the content was still significant (P0.05) lower than that of the control group; muscle T-AOC in stress 2H was significant. Up (P0.05), then descended and significantly lower than (P0.05) control group at the end of the experiment. The activity of SOD and CAT in muscle was significantly lower than that of the treatment after 24 h. The activity of other enzymes was not significantly increased by 2 h (P0.05) 12 h after treatment, and 24 h returned to the pre treatment level. Stress did have a certain effect on the liver tissue structure and antioxidant enzymes of the four finger mackerel young fish, but with the prolongation of the stress time, the liver tissue structure and antioxidant capacity had a certain trend of recovery, while the gills and muscles were aggravated with the time sustained damage, and the structure of the four finger mackerel young fish was formed by.2 stress. The effect of antioxidation system and the effect of anti stress agent mixture to explore the structure of the four fingerling young mackerel, the effect of antioxidant system and the effect of anti stress agents. The study set up the control group (not transported) and the experimental group, and the experimental group included the empty white transport group and the anti stress agent group that was added during the transport process. The anti stress agent mixture (vitamin C: glutamine =1:1) and the experimental group were sampled after 6h transport. The results showed that the cells in the liver part of the group were loosely arranged and the structure of the liver plate was not clear; the liver tissue in the blank transport group was lost in the whole and the structure of the liver plate disappeared. In disorder, the volume of rich mitochondria in mitochondria became larger, the gill fragments in the blank transport group were swollen as a whole, and some of the gill fragments rose because of the excess of red cells. The volume of rich cells in the mitochondria increased and the number increased. The number and size of the white pulp of the spleen were increased relatively, the number and size of the center of melanin macrophage were increased; the blank transport group was increased. The red pulp region of the spleen occupied the majority, the white pulp area was relatively small and the macrophage center decreased. The SOD activity in the liver and muscle was significantly (P0.05) in the control group, and the active value of the liver was much higher than that in the muscle. The CAT activity in the liver (P0.05) showed a blank group of the control group and the muscle in the control group. The content of MDA content in the liver was significant (P0.05) in the blank group and the control group. The value of the muscle in the muscle showed no significant difference (P0.05) between the control group and the mixed group, and the two groups were significantly lower than the blank group (P0.05), while the content of the muscle was much higher than that in the liver; the content of GSH in the muscles was significantly higher (P0.05). There was no significant (P0.05) difference between the control group and the mixed group in the control group, but the two groups were significantly lower than the blank group, and the two groups were significantly lower than the blank group, and the liver T-AOC value was significantly (P0.05) in the blank group and the control group, but there was no significant difference (P0.05) between the mixed group and the blank group (P0), and was significant (P0). .05) was lower than that of the control group; the activity of gill and potassium ATP enzyme in the control group was slightly higher than that in the mixed group (P0.05), and the mixture group was significantly higher (P0.05) than the blank group. The results of the study were four fingers of the fish seedling in the mackerel and the transportation of the mackerel. The effects of.3 transport stress on the antioxidant system of the four finger mackerel young fish and the anti stress agents were found. To explore the effects of transport stress on the liver and muscle antioxidant system of four finger mackerel and the physiological role of anti stress agents. The study set up the control group, the blank group, the vitamin C group and the glutamine group. The determination of the liver and muscle samples were measured in the 2H, 6h and 9h samples carried out in the transport test. The results were as follows: SOD, CAT, MDA, GSH, and T-AOC. results showed that the activity of SOD in the liver and muscles of the blank group was rapidly increased by transport stress, and the existence of anti stress agent was lower than that of the control group; the CAT activity in the liver and muscles of the blank group increased significantly (P0.05) in the blank group, although later, but the activity values were all significant (P0.05). The final value of vitamin C group and glutamine group had no significant difference between the control group and the control group (P0.05) or slightly higher than the control group. Transport stress made the MDA content in the liver and muscle of the blank group increased significantly (P0.05), while the anti stress agent significantly relieved this result; the content of GSH contained different changes in the liver and muscle. The T-AOC value of the liver and muscle in the blank group was significantly higher than that of the control group after transportation stress (P0.05), while the vitamin C group and the glutamine group were slightly higher or equal to the control group. This study analyzed the changes of the antioxidant system and the anti stress effect of the four finger mackerel juvenile fish under the stress of transport. Effect, provide reference for the promotion of artificial culture. The effect of.4 acute low temperature on the tissue structure of four finger mackerel young fish, antioxidant system and the activity of gill sodium potassium ATP enzyme is to explore the effect of acute low temperature stress on the tissue structure, antioxidant system and the activity of gill sodium potassium ATP enzyme in four finger horse mackerel. The study set up a normal temperature (27 + 0.5 degrees C). The group, group 20, 15 C and three temperature gradient were tested, and the normal temperature group was used as control. The liver, muscle, spleen and gill samples were observed and measured at 2h, 6h and 12h respectively. The results showed that the cell vacuoles of the liver increased with the time of treatment at 20, the nucleus deviated, and the staining became shallow. Trend: the liver cells in the 15 C group were arranged in confusion, the structure of the liver was not clear, until the final nucleus dissolves, the whole liver lost its inherent morphology. With the prolongation of the time of cryogenic treatment, the muscle fibers were flexed slightly to the most of the muscle fibers, and the muscle fibers had larger gaps between the fibers and the internal muscles, and the muscle fiber gaps at 15 C were increased. Large, partial fracture until the muscle fiber and the interior are seriously cracked, part of the muscle fibers dissolve and expose the nucleus. With the prolongation of the time of treatment at 20, the number of rich cells in the mitochondria is increased, the end of the gill fragments is slightly enlarged to the whole oedema of the branchial fragments, and the blood vessels and blood sinuses appear in large quantities. Red blood cells, partial gill fragments rose because of too much red blood cells, and a slight shedding of the epidermis of the gill lamellae in 15 C and a few gill lamellae swelled to its water absorption, and the basic morphology of the gill slices was not visible in the liver and muscle, and the trend of the first rise and then decreased in the two temperatures, and ultimately significantly lower than the control (P0.05) was significantly lower than that of the control. In the two temperatures, the presence of CAT in the liver showed a tendency to rise first and then decrease, but in the two temperatures of the muscle, the trend was different, but the active value was significantly lower than that of the control group; GSH in the liver and muscles decreased at two temperatures, and the difference was that the muscle in the muscles at 20 degrees declined significantly. Potential, while in the liver and at 15 degrees of muscle, it was first descended and maintained at a certain trend; MDA in the two temperatures of the liver and in the muscles at 20 degrees centigrade showed a tendency to rise first and then decrease, while at 15 degrees Celsius, the muscle increased first and continued to rise after a period of time; at the two temperature of the liver, T-AOC was at the two temperature of the liver. The muscle in the muscle at 15 C has been declining, but the muscle in the muscle at 20 C is decreasing and descending. The gill Na+-K+-ATP enzyme activity decreases first and then rises at two temperatures, and the difference is at 20 degrees centigrade, and the increase to a certain value remains unchanged under the condition of 15. The results provide references for overwintering management of four finger mackerel fry and improving survival rate.
【学位授予单位】：上海海洋大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S917.4

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