Parentage assignment in hybrid abalones (Haliotis rufescens × Haliotis discus hannai) based on microsatellite DNA markers

Parentage analysis in aquaculture determines genealogical relationships between broodstock and progeny when the parents are unknown. Thus, parentage analysis is a useful tool to establish pedigree reports in molecular‐assisted selection programs. Here, we evaluated 10 heterologous microsatellite markers for parentage assignment in abalone hybrids produced from 43 abalone broodstocks of red abalone (Haliotis rufescens) and Japanese abalone (H. discus hannai). The allele frequencies, exclusion probabilities and broodstock contributions were calculated using CERVUS, PAPA and GERUD software. The polymorphic information content (PIC) values showed that most of the microsatellite loci were highly informative (>0.7) and more than 90% of parentage assignment was possible with a minimum of 5–6 microsatellite markers. Parentage assignment for hybrid and pure‐red progeny showed a better performance than pure‐Japanese progeny. This result could be due to the high level of allele loss in the parental genotypes. In addition, results indicated that only two sires contributed over 80% and 90% of red and hybrid progenies, respectively. This study gives a new molecular tool to support marker‐assisted selection in abalone hybrids produced in Chile.     

皱纹盘鲍眼部组织的显微及亚显微结构观察

采用组织学和电镜的方法,对皱纹盘鲍(Haliotis discus hannai Ino)的眼部组织进行了光镜和电镜观察,以期为进一步从分子生物学角度解析鲍对光照的生理响应机制提供组织学与细胞学基础。结果发现,眼部组织由外至内依次为视网膜色素上皮细胞层、外核层、光感受器内节、内核层、黑色素颗粒沉积层、视觉纤维层。组织表面布满乳头状突起,每一乳状突起的顶端均具有一簇或两簇纤毛环。疏松结缔组织和平滑肌纤维等是组织内的主要成分,结缔组织间分布的胶原纤维等对保持细胞的弹性和韧性具有重要的作用。研究结果显示了鲍的眼部组织在感受和辨识外界光环境因子中的重要作用,也为鲍养殖生产中的光环境因子优化和调控、深入探讨鲍对光照的生理应答机制提供了形态学依据。     

Effects of dietary substitution of fishmeal with the combined dry microalgae,Nannochloropsis oceanica (NO) biomass residue and casein on growth and body composition of juvenile abalone (Haliotis discus)

The effects of dietary substitution of fishmeal with the combined dry Nannochloropsis oceanica (NO) biomass residue and casein on growth performance and carcass composition of juvenile abalone was determined. One thousand two hundred and sixty juvenile abalone were randomly distributed into the 18 L plastic rectangular containers. Five experimental diets in triplicate were prepared. The 28% fishmeal was included into the control (Con) diet. The 25%, 50%, 75% and 100% fishmeal were substituted with the combination of graded levels of dry NO biomass residue and casein, referred to as the NO25, NO50, NO75 and NO100 diets respectively. Finally, the salted sea tangle (ST), Laminaria japonica was prepared to compare the effect of the experimental diets on performance of abalone. The essential amino acids, such as isoleucine, leucine, phenylalanine, threonine and valine tended to increase with dietary substitution of fishmeal with NO biomass residue in the experimental diets. Arginine tended to decrease with dietary substitution of fishmeal with NO biomass residue. Survival of abalone fed the experimental diets was higher than that of abalone fed the ST diet for 16 weeks. Weight gain and specific growth rate (SGR) of abalone fed the NO100 diet were higher than those of abalone fed the all other diets. Weight gain and SGR of abalone linearly increased with dietary substation of fishmeal with NO. Shell length of abalone tended to increase with dietary substitution of fishmeal with the combined dry NO biomass residue and casein. The chemical composition of the soft body of abalone was different among treatments except for moisture content. In conclusion, fishmeal in the diets for abalone could be completely replaced with the combined dry NO biomass residue and casein when the 28% fishmeal was included.     

不同温度下皱纹盘鲍"中国红"与各家系代谢和吸收效率的比较

为了综合评价皱纹盘鲍Haliotis discus hannai红壳色新品系"中国红"(RwRh) 的生理能量学特性,采用实验生态学的方法比较了12、15、18、21、24 ℃条件下,RwRh与皱纹盘鲍不同地理群体(J1Jm、C1Jm、C1Rh、J7Rh和RjRh)的代谢率和吸收效率.结果表明: J1Jm(皱纹盘鲍日本群体♀×日本群体♂)和C1Jm(皱纹盘鲍中国群体♀×日本群体♂)的耗氧率随水温的升高而上升,但超过18 ℃在21 ℃下耗氧率明显下降,RwRh与其半同胞家系(C1Rh、J7Rh、RjRh)的耗氧率拐点出现在21 ℃,说明"中国红"及其半同胞家系更适宜较高温度;各家系排氨率随温度升高的变化趋势与耗氧率基本相同.方差分析显示,在12～24 ℃内,家系对吸收效率的影响未达到显著水平,但在21～24 ℃下"中国红"有较高的吸收效率.     

九孔鲍精子发生的超微结构

利用透射电镜观察九孔鲍（Haliotis diversicolor aquatilis）精子的发生过程。结果表明,九孔鲍精子的发生历经精原细胞、初级精母细胞、次级精母细胞、精细胞和成熟精子5个阶段。其中精细胞的分化可分为5期,主要特点为：Ⅰ期,一些高电子致密颗粒参与顶体的形成;Ⅱ期,核形态由椭圆形到近圆形,再逐渐拉长为长圆柱状;Ⅲ期,核染色质以颗粒状形式凝集,最后向高电子密度均质状态转变;IV期,线粒体逐步融合、体积增大和嵴发达;Ⅴ期,中心体移动及鞭毛形成,胞质逐渐减少。成熟精子为原生型,由头部、中段和尾部组成。通过观察精子发生过程中细胞形态结构的变化,分析了其演变规律,同时与软体动物的其他种类作了比较研究。     

Effects of flow velocity on growth,metabolism and nitrogen and phosphorus budget of Haliotis discus hannai Ino in a recirculating aquaculture system

To study the effects of different flow velocities on growth, metabolism and nitrogen (N) and phosphorus (P) budget of Haliotis discus hannai Ino in a recirculating aquaculture system, abalones (shell length: 45.71 ± 3.09 mm, body weight: 8.62 ± 0.75 g) were cultured at three different flow velocities with four replicates each: high (400 L/hr), medium (300 L/hr) and low (200 L/hr). At the end of the 70 days experiment, the survival rate, food intake and P utilization rate of abalones in the 200 L/hr group were significantly lower than those in any other group (p < .05), whereas the total ammonia nitrogen and nitrite nitrogen concentrations in the water were significantly higher than those in any other group (p < .05). The body weight specific growth rate, food conversion efficiency, abalone‐harvested N and P outputs and N utilization rate were ranked as follows: 300 > 400 > 200 L/hr (p < .05). In the 300 L/hr group, the activities of hexokinase, pyruvate kinase and lactate dehydrogenase were significantly lower than those in the 400 L/hr group, whereas the activity of succinate dehydrogenase was significantly higher than that in the other two groups (p < .05). Aerobic metabolism was the main source of energy acquisition for abalones in the 300 L/hr group. In the 400 L/hr group, the expression levels of manganese superoxide dismutase and thioredoxin peroxidase significantly increased from Day 35 to Day 70, but they still were significantly lower than the values in the 200 L/hr group (p < .05). In the 200 L/hr group, the expression levels of catalase and heat shock protein 70 were significantly higher than those in any other group, and they significantly increased from Day 35 to Day 70 (p < .05). These results indicate that maintaining flow velocity at 300 L/hr will minimize oxidative damage due to deterioration of water quality, reduce the energy consumption required for organisms to resist the impact of water flow, improve the N and P utilization rates of the recirculating aquaculture system, and reduce the cost of culturing abalones.     

Substitution effect of sea tangle (ST) (Laminaria japonica) with tunic of sea squirt (SS) (Halocynthia roretzi) in diet on growth and carcass composition of juvenile abalone (Haliotis discus,Reeve 1846)

Substitution effect of sea tangle (ST) with tunic of sea squirt (SS) in diet on growth and carcass composition of juvenile abalone was determined. One thousand four hundred and seventy abalones were distributed into 21 containers. Six formulated diets in triplicate were prepared. A 200 g/kg ST was included into the ST0 diet. The 200, 400, 600, 800 and 1000 g/kg of ST were substituted with the same amount of tunic of SS, referred to as the ST200, ST400, ST600, ST800 and ST1000 diets, respectively. Finally, Undaria was prepared to compare effect of the formulated diets on performance of abalone. The experimental diets were fed to abalone for 16 weeks. Weight gain of abalone fed the ST400 diet was higher than that of abalone fed the ST0, ST600, ST800 and ST1000 diets and Undaria. Weight gain of abalone fed the formulated diets was higher than that of abalone fed the Undaria. The chemical composition of the carcass of abalone was affected by dietary substitution of ST with tunic of SS. In conclusion, ST could be completely substituted with tunic of SS without retardation in performance of abalone. Abalone fed the ST400 diet substituting 400 g/kg ST with tunic of SS achieved the best growth.     

七彩神仙鱼Vasa基因cDNA的克隆及表达分析

源自DEAD-box家族的Vasa基因是生殖细胞的分子标记之一.本研究克隆了七彩神仙鱼(Symphysodon haraldi)Vasa基因的全长序列,并进行了不同组织的表达分析.结果表明:七彩神仙鱼Vasa基因cDNA序列共2 370 bp,其中5'UTR占123 bp,3'UTR为279 bp,ORF编码655个氨基酸,长1 968 bp.氨基酸序列同源性分析表明七彩神仙鱼的Vasa基因与尼罗罗非鱼(Oreochromis niloticus)的同源性最高.半定量分析表明,Vasa基因在成熟七彩神仙鱼的性腺中特异表达,在其它组织中无表达信号.qRT-PCR结果显示,Vasa在七彩神仙鱼早期胚胎发育阶段及出膜后50日内均有表达.在受精卵至囊胚期阶段,Vasa基因表达持续增加,在囊胚期至孵化期阶段,表达持续降低.在仔鱼阶段,Vasa分别在25日龄和40日龄出现了最低和最高表达量.繁殖前后比较发现,繁殖前的精巢组织中Vasa表达量比繁殖后的表达量低,而卵巢恰好相反.本研究结果可为研究七彩神仙鱼的性分化、生殖细胞分子标记及其发育提供参考.     

不同卵色皱纹盘鲍(Haliotis discus hannai)幼鲍的生长与免疫比较

随机选取100只经过480 d养殖的3♀×10♂绿卵全同胞家系子一代(F1)的皱纹盘鲍(Haliotis discus hannai)幼鲍为实验材料,进行雌雄、卵色分类后,测量所有样本总重、壳长、壳宽等生长指标,并测定不同卵色幼鲍的超氧化物歧化酶(SOD)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、过氧化氢酶(CAT)和溶菌酶(LZM)的活性,进行单因素方差分析(One-way ANOVA).结果显示,在生长方面,不同卵色幼鲍的各生长指标无显著差异(P＞0.05),但绿卵幼鲍的各生长指标均高于其他3种卵色;在免疫酶活性方面,4种卵色幼鲍在SOD和CAT活性上无显著差异(P＞0.05),绿卵幼鲍的AKP、ACP和LZM 3种酶的活性与棕色和棕红色卵幼鲍存在显著差异(P＜0.05),而与棕绿色卵幼鲍无显著差异,其酶活性从高到低按卵色排列依次为绿色、棕绿色、棕色和棕红色.上述研究结果可为利用卵色选育皱纹盘鲍高产抗逆新品系提供依据.