【作者】 梁淼；

【导师】 董双林；

【作者基本信息】 中国海洋大学 ， 水产养殖， 2010， 博士

【摘要】 本文详述了个体生长差异的研究进展及青刺参和红刺参的比较研究进展,并通过一系列的室内实验分析了导致刺参个体生长差异如此之大的原因。研究结果总结如下：1不同放养密度和初始规格对刺参个体生长差异的影响。实验设置5、10、20、30、40、50头/100L 6个密度处理(分别记为D5、D10、D20、D30、D40和D50)实验持续50d；之后,再将实验刺参分为大、中、小3个规格处理(分别记为H、M和L)和1个3个规格混合处理(记为HET)继续实验50d。实验结果表明,(1)在食物持续稳定供应的情况下,D5-D30范围内刺参随放养密度的增大其个体生长变异也增大,但当密度大于D30后其个体生长变异不再明显增加。刺参密度与生长的关系符合阿利氏群聚原则,D20是刺参最佳的放养密度,此密度下刺参平均生长率最大,其蛋白和脂肪积累也最多。(2)刺参具有明显的社会等级行为,优势者的大个体(H)抑制了劣势者较小个体(M和L)的生长,劣势者较小个体甚至会出现负生长。(3)出现较大个体生长变异刺参群体,在将大规格的刺参分离出去后,中等个体M会出现生长补偿现象,而较小的个体L会出现生长退尝现象。较小个体的刺参活动减少,摄食率明显减小,用于生长的能量比例减小,而用于呼吸和排泄的能量比例增大。(4)混合规格处理HET的刺参个体生长变异超过70%,其是密度、社会等级行为和遗传三类因素共同作用的结果,而且遗传因素大于社会等级行为因素的作用。2刺参个体生长差异的生理学研究。研究了小个体刺参在大个体刺参存在情况下生长、呼吸和排泄、Na+/K+-ATPase、丙酮酸激酶和己糖激酶活性的变化。实验首先将2000头4-6g的刺参在2200L的大水槽中饲养60d,这些饲养的刺参出现明显的个体差异,之后,再将实验刺参分为大、中、小3个规格处理(分别记为H、M和L)和1个3个规格混合处理(记为HET)。4种处理实验60d后刺参又出现个体分化,大规格组中出现大规格刺参(Hb)和小规格刺参(Hs),中规格刺参组中出现大规格刺参(Mb)和小规格刺参(Ms)、小规格刺参组中出现大规格刺参(Lb)和小规格刺参(Ls),混合规格组刺参分为大中小三种规格(HETb, HETm和HETs)。实验结果表明,在食物持续稳定供应及大规格刺参“胁迫”下,小规格刺参的特定生长率均出现负值,相应的耗氧率和排氨率显著低于大规格刺参(P<0.05)。同时,在大规格刺参“胁迫”下,小个体刺参在生理功能上呈现出显著的变化,如Na+/K+-ATPase活力显著升高(P<0.05),己糖激酶和丙酮酸激酶活力显著下降(P<0.05),小个体刺参糖酵解作用被抑制,能量代谢降低,生长受到抑制。然而,在整个实验过程中并没有观察到大个体刺参对小个体刺参明显的攻击行为,因此这种“胁迫”作用对小个体刺参的生理功能造成显著的影响。3单个体饲养条件下刺参的个体生长差异。刺参养殖群体通常具有异常大的个体生长变异,本研究通过个体隔离饲养方法,排除了社会等级行为和放养密度等因素的影响,探讨刺参个体生长差异如此大的原因。实验结果表明,经过100d的隔离饲养,刺参个体之间在生长上出现明显的个体差异,它们的个体特定生长率相差近3倍(0.40%-1.01%),其体重变异系数从12.04%扩大至40.51%。刺参初始体重与其后继生长、摄食率和食物转化效率具有密切的正相关性(P<0.05)。能量概算表明,初始体重较小的个体呼吸消耗的能量比例更高一些(个体间差异近4倍),用于生长的能量比例更少一些(个体间差异近2倍)。群体饲养条件下刺参的CV小于单独隔离饲养条件下的CV这一事实表明,群体饲养的刺参个体间的相互作用也影响着刺参的个体生长变异。本研究证实,刺参个体间存在显著的生物学差异,这是其群体生长变异很大的主要原因,这为刺参养殖品种的选育奠定了理论基础。在刺参养殖实践中,适当控制刺参的放养密度并经常性地进行规格分级对于提高养殖群体的平均生长速度和规格均一性将会是十分有效的。4社会等级因素对刺参生长差异的影响。实验初始刺参依大、小两种规格分成2类,即大规格刺参(H)体重为10-12g,小规格刺参(L)体重为3-4g。按大规格与小规格分配比例分为5个处理组,即大规格刺参：小规格刺参分为2:18、4:16、6:14、8:12、10:10头/槽,分别记为B、C、D、E和F组,对照组为20头小规格刺参,记为A组。结果表明,刺参表现为明显的线性等级制度,即H规格刺参处在第一等级(dominant)、L-big规格处在第二等级(Lb)和L-small规格处在第三等级(subordinate, Ls)。随着占主导地位的优势者大个体(H)刺参数量的增多,劣势者较小个体(Ls)的摄食、生长呈现下降的趋势,出现生长退尝现象。在没有大个体(H)刺参“胁迫”下,对照组(A)中Lb规格刺参的SGR最高,达到1.25%d-1,显著高于各处理组(P<0.05)。H和L规格刺参的体重变异系数实验过程中存在显著的差异,且体重变异系数与时间呈现显著的正相关。实验结束时,对照组A的变异系数最高,达到61.40%。刺参的体组成存在一定的变化规律,随着H规格刺参数量的增加,Lb和Ls规格刺参的粗脂肪和粗蛋白含量减少,而粗灰分含量增加。本研究的结果表明,在刺参养殖过程中,通过定期规格分级可减轻大个体刺参对小个体刺参的“胁迫”作用,有利于保证规格的均一性,进而提高刺参养殖的生产效率。5社会等级因素存在下刺参个体生长差异的生理学研究。通过测定社会等级行为下优势个体和劣势个体刺参的呼吸排泄、代谢酶和消化酶的变化,分析等级制度胁迫下不同等级刺参的生理响应模式。实验初始刺参依大、小两种规格分成2类,即大规格刺参(H)体重为10-12g,小规格刺参(L)体重为3-4g,按大规格与小规格的分配比例分为5个处理组,即大规格刺参：小规格刺参为2:18、4:16、6:14、8:12、10:10头/槽,分别记为B、C、D、E和F组,对照组为20头小规格刺参,记为A组。结果表明,在生理功能上刺参表现为明显的线性等级制度,即H规格刺参处在第一等级(dominant, H)、L-big规格处在第二等级(Lb)和L-small规格处在第三等级(subordinate, Ls)。刺参耗氧代谢的等级高低顺序为,H规格>Lb规格>Ls规格,随着等级胁迫的增强,Lb和Ls规格刺参的代谢率呈现下降的趋势。高等级刺参(H、Lb)的代谢生理和消化生理活跃,表现为较高的代谢酶活力(丙酮酸激酶、己糖激酶、乳酸脱氢酶、乳酸含量和ATP含量)和消化酶活力(蛋白酶、淀粉酶和脂肪酶活力),低等级刺参(Ls)在高等级刺参的“胁迫”下,其代谢生理和消化生理紊乱,表现为代谢酶和消化酶活力的显著降低(P<0.05)。等级制度下刺参生理机制的研究结果表明,通过规格分级可减轻由社会等级因素引起的生理功能胁迫,有益于减小个体间的生长差异。6单养和群养下红刺参和青刺参个体生长差异。比较研究了单养和群养下红刺参和青刺参的生长、摄食和能量分配,并对红、青刺参的个体生长变异进行了比较。青刺参和红刺参先分成2个饲养组,即单养组(A)和群养组(B),养殖60d后再进行4个处理,即一直单养组(Aa)、一直群养组(Bb)、单养后群养组(Ab)和群养后单养组(Ba),再养殖60d。实验结果表明,多数处理中红刺参的生长快于青刺参(P<0.05),其主要原因是红刺参的摄食率高,摄食能中用于呼吸的比例(R/C)较低；多数处理中红刺参与青刺参的摄食能用于排泄的比例(U/C)差异不显著(P>0.05),但前者摄食能中未吸收的能量比例(F/C)高于后者；红刺参的体重变异系数(CV)显著低于青刺参(P<0.05)。本实验中刺参个体单养方式(Aa)消除了动物个体间的“胁迫”作用,致使其个体的平均生长速度快于群养组(Bb)。个体单养处理的刺参仍表现出较大的个体变异,而且一直群养Bb与一直单养Aa的刺参的体重变异系数的差值小于它们一直单养的体重变异系数,表明造成刺参生长变异的遗传因素大于个体间“胁迫”因素。刺参个体体重变异大的处理其平均生长速度较慢。本研究表明,在刺参养殖生产中适时进行规格分级有助于提高生产量,对刺参进行选育具有良好的遗传学基础。7单养和群养下红刺参和青刺参个体生长差异的生理学研究。本文通过测定单养和群养下红刺参和青刺参体壁和肌肉束的丙酮酸激酶(PK)、己糖激酶(HK)、乳酸脱氢酶(LDH)、乳酸含量(LD)和苹果酸脱氢酶(MDH)活性,以分析红、青刺参能量代谢的生理响应机制。结果表明：不同处理的红刺参体壁和肌肉束中的PK、HK和MDH酶活性均高于青刺参,在Ab处理中表现为差异显著(P<0.05),但红、青刺参肌肉束中的LDH酶活力差异不显著(P<0.05),而且在红、青刺参的体壁中均未检测出LDH酶；分组后(Ab组和Ba组)的红刺参的LDH酶活力高于没分组(Aa组和Bb组),青刺参的酶活力变化与红刺参相反；在各处理组中,红、青刺参LD含量均在Ab组中最高,且红刺参体壁、肌肉束中的LD含量均高于青刺参,尤其在体壁中表现为差异显著(P<0.05)。本研究从生理机制上证明适时分组有利于刺参在生理上做出积极响应,进而满足生长和代谢的需求,通过降低刺参的个体差异来提高生产效率。更多还原

【Abstract】 1 The present studies deal with the intra-specific effects of sea cucumber Apostichopus japonicus with unlimited food resources, especially the effects of stocking density on growth variation of the animal and energetic changes of small individuals under the stress of large individuals. The results showed that with the initial body weight of 5.12±0.09-6.11±0.26g of sea cucumber among the densities of 5,10,20,30,40 and 50 ind./100L, the density of 20 ind./100L was the optimum stocking density because of its highest specific growth rate, crude protein content and crude lipid content in tissue. Individual growth variation of A. japonicus increased with the increase of stocking densities, whereas no significant differences in the variation were found when the density was over 30 ind./100L (P>0.05). The low-weight individuals under the stress of heavy-weight individuals exhibited obvious changes in energetics, such as lower ingestion rate, lower energy deposited as growth but higher respiration and excretion. The coefficient of variation (CV) in growth of the animals was over 70% due to simultaneous action of aggression and maybe a factor of chemical mediator, and led to significant changes in energetics of small size individuals.2 The present studies were conducted to elucidate the effects of large individuals of sea cucumbers Apostichopus japonicus Selenka on the physiological performance of small individuals of sea cucumbers by measuring growth, respiration, excretion, Na+/ K+-ATPase, pyruvate kinase and hexokinase. After certain time of cultivation an obvious size differences were seen among the sea cucumber individuals in different treatments. The results showed that with sustainable food supply and under the "stress" of large-size A. japonicus, the SGRs of the small-size ones became negative in values, their oxygen consumption rates and ammonia excretion rates were significantly lower than those of the big-size A. japonicus (P<0.05); The small-sizes individuals under the "stress" of big-size individuals exhibited obvious changes in physiological performance, such as, the Na+/K+-ATPase activity enhanced significantly (P<0.05), HK activity and PK activity reduced significantly (P<0.05), which implies that the glycolysis pathway of small-sizes individuals might be inhibited to some extent by the "stress" of big-sizes individuals. The aggression behavior of A. japonicus was not seen obviously during the experiment. However, the small-size individuals under the "stress" of big-size individuals exhibit some changes in physiological performance.3 The exceptionally large individual growth variation has been previously recognized in several sea cucumber cohorts. However, there is a lack of information regarding the mechanism of such individual differences. In this study, the sea cucumber Apostichopus japonicus (Selenca) was reared individually in order to eliminate possible effects of social interaction, stocking density, etc. The results showed that there were substantial differences in growth among the sea cucumber individuals during the 100-day experiment. The special growth rate of the sea cucumber individuals differed by up to three folds (from 0.40% to 1.01%), and the coefficient of variation in body weight increased from 12.04% to 40.51%. The final body weight, food intake and food conversion efficiency for each sea cucumber were generally positively correlated with their initial body weight (P<0.05). Energy budget of the animals showed that the food energy spent on respiration was much greater (about four folds) but energy deposited to growth was much less for (initially) smaller than for larger A. japonicus. The present result implies that there are obvious genetic differences among the sea cucumber individuals, largely accounting for the individual growth variation of the cohort sea cucumber. These results will provide some basic data for promoting selective breeding and farming of the sea cucumber. 4 The present study deals with the effects of social hierarchy on individual growth variation of sea cucumber. All cohort sea cucumbers were size-graded and divided into two groups, i.e. the heavy-size (H) A. japonicus group (initial body weight 10-12g) and the light-size (L) A. japonicus group (initial body weight 3-4g). Six social hierarchy treatments of A. japonicus, i.e. the heavy-size (H) A. japonicus:the light-size (L) A. japonicus 0:20,2:18,4:16,6:14,8:12 and 10:10 ind/160L fiberglass tank (represented as A, B, C, D, E and F, respectively) were used in this trial. The results showed that:(1)there were obvious social structures among individuals A. japonicus, and the heavy-size A. japonicus was the first rank (dominant, H), the L-big size was the second rank (Lb) and the L-small size was the third rank (subordinate, Ls). (2)The Ls A. japonicus showed a negative growth under the double "stress" of the H and Lb A. japonicus. The maximum value of SGR occurred in the Lb A. japonicus of A treatment and reached 1.25%d-1, which was significantly higher than those in other treatments (P<0.05). (3)The CVs for the body weight of H and L A. japonicus were significant difference during the 80-day’s trial, and the CVs for the body weight was positively correlated to the experiment time (P<0.05). The maximum value of CVs for the body weight occurred in the A treatment and reached 61.4%. (4)Under the 80-day’s experiment, the biochemical composition of H, Lb and Ls A. japonicus exhibit obvious changes. With the number of H A. japonicus increasing, the crude fat contents and the crude protein contents of Lb and Ls A. japonicus decreased, and the crude ash content increased. So in sea cucumber aquaculture practice, it will be effective to alleviate the "stress" of the dominant individual, improve production and size uniformity by size grading periodically.5 The present studies were conducted to elucidate the effects of social hierarchy on the physiological performance of dominant and subordinate individuals of sea cucumbers Apostichopus japonicus Selenka by measuring respiration, excretion, metabolic enzymes and digestive enzymes. All cohort sea cucumbers were size-graded and divided into two groups, i.e. the heavy-size (H) A. japonicus group (initial body weight 10-12g) and the light-size (L) A. japonicus group (initial body weight 3-4g). Six social hierarchy treatments of A. japonicus, i.e. the heavy-size (H) A. japonicus:the light-size (L) A. japonicus 0:20,2:18,4:16,6:14,8:12 and 10:10 ind/160L fiberglass tank (represented as A, B, C, D, E and F, respectively) were used in this trial. The results showed that:(1)there were obvious social structures among individuals A. japonicus in the physiological function, and the heavy-size A. japonicus was the first rank (dominant, H), the L-big size was the second rank (Lb) and the L-small size was the third rank (subordinate, Ls). (2)The oxygen consumption rates and ammonia excretion rates were positively correlated to social hierarchy of A. japonicus (P<0.05), and were shown in descending order as followed:H>Lb>Ls; Meanwhile, with the "stress" of the number of H-size A. japonicus increasing, the oxygen consumption rates and ammonia excretion rates of the Lb and Ls A. japonicus exhibited significantly decreased (P<0.05). (3)The H and Lb size A. japonicus were all in the physiological activity stage because of their highest activities of metabolic enzymes and digestive enzymes. However, the Ls size A. japonicus under the double "stress" of H and Lb size individuals exhibited a metabolic and digestive disturbance period, such as the lowest activities of metabolic enzymes and digestive enzymes. So in sea cucumber aquaculture practice, this study reconfirmed that it will be effective to alleviate the "stress" of the dominant individual in the physiological functions, and it will be necessary to improve production and size uniformity by size grading periodically.6 Specific growth rate, food intake rate and energy allocation of Red and Green Apostichopus japonicus were studied in both individual housed and group housed treatments in order to compare the growth performances of the two variants and to elucidate quantificationally the mechanism of their individual growth variation. The results showed that the growth rate of Red variant was significantly higher than that of Green variant in most treatments after 120-day’s trial (P<0.05). Red variant had higher food intake rate and less percentage of energy lost in respiration than Green variant, and the coefficient of variation for body weight of the former was significantly lower than that of the latter (P<0.05). The value differences of the coefficient of variation for body weight of Red and Green variants between the continuous individual-housed treatment and continuous group-housed treatment were 11.56 and 19.32, respectively. The value differences were less than the corresponding values in the continuous individual-housed treatment (25.90% and 41.91%, respectively). The results indicate that the inherent factor is more important than the intra-specific competition in the contribution to the individual variation of A. japonicus in growth.7 In this study, we determined the activities of pyruvate kinase (PK), hexokinase (HK), lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) and the content of lactic acid (LD) in the body walls and muscles of Red and Green variants of sea cucumber Apostichopus japonicus respectively, which were reared under individual and group housing in order to analyze the effects of different culture modes on the physiological performance of sea cucumbers. The results showed that under different treatments, the activities of PK, HK and MDH in the body wall and muscle of Red variants were higher than those of Green variants, especially in the Ab treatment (P<0.05). However, the LDH activity in the muscle of Red variants was slightly higher than that of Green variants, but the difference was not significant (P>0.05). The LDH activity of Red variants in the Ab and Ba treatment was higher than that of Red ones in the Aa and Bb treatment, but Green variants were contrary. The maximum value of the LD content of Red and Green variants occurred in the Ab treatment, and the LD content of the body walls and muscles of Red individuals was higher than that of Green, especially in the body walls (P<0.05). So in sea cucumber aquaculture practice, it reconfirmed that the re-grouped housing mode will be propitious to the physiological positive responses, and it will be effective to improve production and size uniformity by meeting the needs of higher growth and higher metabolize.更多还原