【作者】 张建设；

【导师】 苏永全；

【作者基本信息】 厦门大学 ， 海洋生物学， 2007， 博士

【摘要】 曼氏无针乌贼（Sepiella maindroni）曾是舟山渔场四大海产之一。上世纪九十年代以来,由于过度捕捞,自然资源遭到极大破坏。进行人工繁育是对曼氏无针乌贼资源进行保护和恢复的有效方法。本研究主要包括人工养殖条件下的曼氏无针乌贼的繁殖生物学、环境因子耐受性、生长特性、营养成分分析与评价,以及曼氏无针乌贼血细胞的分类及免疫特性等研究,为曼氏无针乌贼的人工增养殖和育种提供理论依据,并为曼氏无针乌贼的规模化生产奠定基础。在人工养殖条件下,曼氏无针乌贼的繁殖习性发生了重大改变,养殖的曼氏无针乌贼由原来每年的单次到两次的繁育活动。曼氏无针乌贼的卵巢发育与温度有明显相关,较高的温度（21.5-23.5℃）可促进卵巢的发育。曼氏无针乌贼5月份成熟个体怀卵量在565-4430粒,一般在1015-2487粒,平均2021粒;10月份成熟个体怀卵量870-4354粒,一般在1705-2937粒,平均2399粒。怀卵量随胴长和体重的增大而升高;绝对生殖力和相对生殖力也与胴长和体重呈线性正相关关系。对曼氏无针乌贼产卵附着基进行筛选,选择草绿色聚乙烯有结节网片作为附着基材料,对附着基进行结构优化。对曼氏无针乌贼胚胎发育进行显微观察。曼氏无针乌贼受精卵发育生物学零度（C）为6.53℃,受精卵发育至刚出膜幼体的有效积温（K）值为397.13℃·d。曼氏无针乌贼胚胎发育的最适宜温度范围为18.0-24.0℃,最适宜盐度范围为20.00-30.00,最适宜pH范围为7.0-9.0。曼氏无针乌贼仔稚幼体存活和生长最适温度范围为18.0～26.0℃,盐度的适宜范围为20.00-30.00,pH的适宜范围为7.5-8.5。曼氏无针乌贼幼体对活饵料的选择性极强,轮虫可作为曼氏无针乌贼的开口饵料。在饵料充足的条件下,曼氏无针乌贼仔稚幼体体长和体重增长较快,体长的总平均日增长率为2.86%,最高增长率为3.98%,最低增长率为1.93%;体重的总平均日增长率为8.61%,最高增长率为10.92%,最低增长率为6.76%。体重（W）与体长（L）的关系为:W=0.3038L^2.909,R²=0.9698,为均匀增长类型。幼体至成体生长阶段,体长和体重的日增长率降低,且表现出雌雄差异。曼氏无针乌贼雌体体长（L）与日龄（D）的关系为:L_?=0.1936D-9.416,R²=0.9928;雄体体长（L）与日龄（D）的关系为:L_?=0.161D-6.266,R²=0.9775。曼氏无针乌贼雌体体重(W_?)与日龄（D）的关系为:W_?=4.2967D-289.12,R²=0.9926;雄体体重(W_?)与日龄（D）的关系为:W_?=3.9772D-264.44,R²=0.9933。体重与体长的关系为:雌体的体重W_?=3.1256L_?^1.6398,R²=0.9918:雄体的体重W_?=2.0268L_?^1.8115,R²=0.9846。对曼氏无针乌贼的生物学性状进行测定,结果表明,曼氏无针乌贼的胴长与胴宽、壳长与壳宽都不是简单的倍数关系,是随生长阶段和性别的不同而变化的。曼氏无针乌贼胴长和内壳长度之间存在线性关系,相关系数大于0.99,因此内壳长度可作为曼氏无针乌贼生长的精确测量指标。测定了养殖曼氏无针乌贼、野生曼氏无针乌贼和金乌贼肌肉生化成分。结果表明:养殖曼氏无针乌贼肌肉生化成分（鲜重百分比）水分含量为75.51±1.92%、蛋白质16.45±0.62%、脂肪1.16±0.068%、灰分3.51±0.30%、糖4.27±0.31%;比能值5.08kJ/g及E/P30.88kJ/g。肌肉中含有18种氨基酸,总量为15.41%（湿重百分比）,9种人体必需氨基酸总量是6.16%,必需氨基酸占氨基酸总量比例为39.97%:呈味氨基酸含量为45.42%;氨基酸评分为82分,蛋白营养价值高。缬氨酸,色氨酸为限制性氨基酸。养殖曼氏无针乌贼的高度不饱和脂肪酸二十五烯酸（EPA）和二十二碳酸（DHA）的含量最高,为5.0mg/g。样品富含钙、镁、锌、铁、磷等矿物质元素。曼氏无针乌贼肌肉是具有高蛋白、低脂类、低比能值、低E/P值等特点的优良食品。采用三种染色方法对曼氏无针乌贼血细胞进行染色,结果表明曼氏无针乌贼血细胞可分为两类:颗粒血细胞和透明血细胞。颗粒血细胞在体外有较强的吞噬能力,透明细胞的吞噬能力不明显。血细胞的密度随乌贼状态的不同而不同,饥饿乌贼的血细胞密度显著降低。正常乌贼血细胞抗菌活力U_a为0.352,饥饿组乌贼的血细胞抗菌活力U_a为0.287,患病组乌贼的血细胞抗菌活力U_a为0.376。正常乌贼血细胞溶菌活力U_L为0.225,饥饿组乌贼的血细胞溶菌活力U_L为0.178,患病组乌贼的血细胞溶菌活力U_L为0.316。活性氧的产生都随细菌浓度的增加而增加,在本实验设置的的最大细菌浓度10⁸ cells/ml时,三种状态乌贼血细胞活性氧的产生量都处于最高值。更多还原

【Abstract】 Sepiella maindroni once was one of the four famous marine products of Zhoushan fishing ground in 1970s.But since 1990’s,the nature resource of S. maindroni have been severely damaged because of overfishing.Artificial propagation is an effective way for protecting and resuming the resource of S.maindroni.The contents of this thesis include propagate biology,tolerance of larvae to environmental conditions,growth characteristics,analysis and evaluation of nutrition composition and haemocytes immune function of cultured S.maindroni.This research aimed at offering theory foundation to the aquaculture;laying a foundation to the large-scale production and ecological restoration of S.maindroni resource in Zhoushan fishing ground.The propagate biological characters of this animal has been changed in cultured condition,S.maindroni might spawn twice a year.There were distinct relationship between the growth of ovary and temperature,higher temperature（21.5-23.5℃） accelerate the growth of ovary.The individual fecundity ranged from 565 to 4430 grain（mean=2021 grain） for individuals matured in May,and from 870 to 4354 （mean=2399 grain） for individuals matured in October.It seemed that the individual fecundity increase with the accretion of length or weight,and the individual absolute fecundity and relative fecundity had a linear positive relationship with the length and weight.The grass green polyethylene nodosity net,polyethylene nodosity strands and bamboo chopsticks were tested as the substance for adhering material of spawning, and the former seemed the optimum one.Microscopic observation on the development of embryo of S.maindroni was carried out.The biological zero temperature of embryonic development was 6.53℃, and the effective accumulated one was 397.13℃·d.The optimum temperature,salinity and pH ranged respectively from 18.0 to 24.0℃,20.0 to 30 and 7.0 to 9.0 for the fetation of S.maindroni.The optimum temperature,salinity and pH for survival and growth of S.maindroni larvae were 18.0 to 26.0℃,20.00 to 30.00 and 7.5 to 9.5, respectively.S.maindroni larvae showed high degree of selectivity for alive bait.Rotifer could be used as the starter feed for this tested larvae.With enough feed,the length and weight of S.maindroni increase rapidly during larvae stage.In this period,the growth rate of length and total body weight was 2.86%per day and 8.61%per day, respectively.The relationship of length and body weight seemed exponential.These relationships could be expressed by a curvilinear correlation as follows: W=0.3038L^2.909,R²=0.9698.S.maindroni generally show even growth pattern in this period.During the period from larvae to mature,the growth rate decreased and showed difference in female and male.The relationship between length and day age for female individuals was L_♀=0.1936D-9.416,R2=0.9928,and L_♂=0.161D-6.266, R²=0.9775 for males,those between body weight and day age for female and male were W_♀=4.2967D-289.12,R²=0.9926 and W_♂=3.9772D-264.44,R²=0.9933 respectively.It was recorded that ratioes of body weight to length was W_♀= 3.1256L_♀1.6398,R²=0.9918,in female;and W_♂=2.0268L_♂^1.8115,R²=0.9846 in male.Several biological characters such as length,body width,cuttlebone length and width of S. maindroni were determined.The results showed that there were not simple positive linear relations between length and body width or shell length and width.The relationships were changed following the developing stage and sex.The relationship between length and shell length was linear,the correlation coefficient was higher than 0.99.Thus tshell length could be used for evaluating growth.Biochemical compositions of cultured S.maindroni,wild S.maindroni and S. esculenta were analyzed.The nutritive composition（%,wet） of cultured S.maindroni were that:muscle moisture 75.51±1.92%,protein 16.45±0.62%,fat 1.16±0.068%, ash 3.51±0.30%and sugar 4.27±0.31%.The content of energy was 5.08kJ/g,E/P was 30.88kJ/g.18 common amino acids were found in muscle and 9 of them were essential amino acids for human needs.The total content of amino acids was 15.41% （wet）;that of the essential amino acids was 6.16%;the percentage of essential amino acids in total amino acids（E/TA） accounted for 39.97%.The delicious amino acids, i.e.Asp,Glu in total amino acids were 45.42%.The protein nutrient value was high, with the amino acids score（AAS） 82,the first and second limited amino acids were valine and tryptophane.In cultured S.maindroni muscle,the contents of EPA and DHA were also high;the percentage of highly unsaturated fatty acids（HUFA） reached 5.0mg/g.Minerals such as Ca,Mg,Zn,Fe and P in the tested samples seemed plentiful.The result showed that cultured S.maindroni contained higher protein, lower fat,low energy and E/P.Three kinds of histological staining methods were used to study the morphology of haemocytes in cuttlefish S.maindroni.Two types of haemocytes were identified: hyalinocytes and granulocytes.The granulocytes had strong phagocytosis ability to Staphylococcus aureus and Escherichia coli.The haemocytes densities of starved S. maindroni were lower than that of control or sicken.Antibacterial activities of the control group,the starved and the sicken ones were 0.352,0.287 and 0.376 respectively,bacteriolysis of the above groups were 0.225,0.178 and 0.316.Oxygen productions increased with bacteria density,and reached highest when bacteria density was 10⁸cells/ml.更多还原