基于AMMI模型分析大菱鲆选育家系基因型与环境互作效应

为选育具有普适性的优良家系和适于某一特定环境的优良大菱鲆家系,实验以大菱鲆选育F2的10个优良家系为材料,利用随机区组设计,将其推广到5个不同的实验点进行养殖实验,采用主效可加互作可乘模型(AMMI)分析方法,分析大菱鲆选育家系基因型与环境互作效应。结果显示,家系和环境互作效应(G×E)的平方和占总平方和的10.82%,达到极显著水平(P0.01),其对产量差异的影响大于家系基因型间对产量差异的影响,为家系效应的1.44倍;综合双标图AMMI模型分析和稳定性参数分析,家系G5和G8的产量较高,分别为844.796和868.888 g,稳定性参数也最小,分别为1.154 975和2.668 016,属于高产、稳产、广适应性较好的家系,其基因型适于新品系(或新品种)选育;通过双标图分析,5个实验点可被分为3组,E3为一组,E5为一组,E1、E2和E4为一组。家系G6的产量为851.768 g,在环境E3条件下最高产,适于在E3条件下推广;家系G10的产量为911.664 g,在环境E5条件下最高产,适于在E5条件下推广;而G2的产量为784.764 g,在环境E1、E2和E4条件下最高产,适于在环境E1、E2和E4条件下推广。     

Characterization of a megalocytivirus from cultured rock bream,Oplegnathus fasciatus (Temminck & Schlege), in China

Megalocytivirus belongs to the Iridoviridae family and is known to affect finfish. Megalocytivirus epizootics have been reported to occur in several cultured fish species in China; however, no megalocytivirus associated with rock bream, Oplegnathus fasciatus (Temminck & Schlege), has been documented. In this study, we characterized for the first time a megalocytivirus, rock bream iridovirus (RBIV)‐C1, detected in cultured rock bream in a fish farm in China that had been inflicted with a high‐mortality disease outbreak. Sequence analysis of three conserved genes showed that RBIV‐C1 shares over 90% overall identities with a number of known megalocytiviruses. Electron microscopic examination revealed RBIV‐C1 as hexagonal particles similar to those reported for megalocytiviruses. In vivo infection study indicated that, following inoculation into rock bream, RBIV‐C1 induced 100% mortality and upregulated the expression of Mx, IL‐1β and IL‐8. The infected fish exhibited pathological signs similar to those observed in naturally diseased fish. Furthermore, studies in a turbot (Scophthalmus maximus L.) model indicated that RBIV‐C1 induced acute infection in turbot that led to 100% mortality. These results indicate that RBIV‐C1 is highly virulent to rock bream as well as turbot and that RBIV‐C1 is closely related to a number of previously reported megalocytivirus and likely a genetic variant of the latter.     

Effects of two diets on lipid composition and reproductive performance of brill (Scophthalmus rhombus) eggs

The effects of two diets with different lipid composition (squid Loligo gahi and one commercial pellet for breeders) on reproductive performance and egg lipid composition of brill (Scophthalmus rhombus) were compared. A total of 36 adult brill weighing 1482 ± 432 g were randomly distributed in the experimental tanks. A flow‐through system composed of four tanks with 4.2 m² of bottom area (5.0 m³ × 1.2 m water depth) was used. The experiment lasted for approximately 5 months (168 days). From the four mature females fed squid, three ovulated. Five females fed pellets matured and ovulated. Total ovulations were of 10 and 44, by the three females fed squid and the five ovulating female fed pellets respectively. Egg viability was higher, although not significantly, for females fed pellets. This indicates that pellets could be a better food for breeding females. In general, the egg total lipid content and the lipid classes composition did not seem to be affected by diet. In contrast, results indicate that not only diets but also individual females, regardless of the diet consumed, are determinant for egg fatty acid composition, clearly modulating their composition.     

大菱鲆家系生长和耐热性状对比分析

利用40个大菱鲆(Scophthalmus maximus)家系进行耐热性实验,测定相应生长和耐热性状指标,对耐热性状进行方差分析,并分析生长与耐热性状的相关情况。结果表明:在各家系生长前期,快速生长家系6个,生长较快家系26个,生长速度一般家系8个。在生长后期阶段,快速生长家系9个,生长较快家系28个,生长速度一般家系3个,仅有8号家系一直为快速生长家系。在耐热性状方面,各家系耐热性平均值在1 367.30～836.30℃.h之间,总体平均值为1 113.00℃.h。高耐高温家系为8号、33号和37号家系,其中33号家系耐热性数值最高,其耐热性平均值为1 367.30℃.h,不耐高温家系为3号、9号、16号、38号和40号家系,其中38号家系耐热性数值最低,其耐热性平均值为836.30℃.h。方差分析结果显示,不同家系间耐热性存在极显著差异(P<0.01),相关分析结果表明,各家系不同生长阶段与耐热性呈现不同的相关性。旨在探明生长性状与耐热性状的相关性,为大菱鲆的耐热家系选育奠定研究基础。     

大菱鲆选育家系雌、雄群体的生长发育差异

采用全长/体长、体高/体长、全长/体高和体质量为评价指标,分别对9~33月龄大菱鲆雌、雄群体在生长发育过程中体型的动态变化及生长性能差异进行了比较。大菱鲆雌、雄群体形态比较结果显示,雌、雄群体的全长/体长、体高/体长、全长/体高在不同发育阶段呈现特有的变化,但在雌、雄群体间,除33月龄的体高/体长和全长/体高差异达到显著水平外,3项指标在不同阶段没有差异或差异不显著,即可以认为,对于统计初始体质量相同或相近的大菱鲆雌、雄群体,全长/体长、体高/体长和全长/体高3个形态比例指标是随发育时间序列同步变化。基于Logistic模型对大菱鲆雌、雄群体生长性能的比较结果显示,雌性群体的拐点月龄、拐点体质量和最大月增重均大于雄性群体;对于雌性和雄性群体的瞬时增长率,除在9~13月龄雌性群体稍低于雄性群体外,13~33月龄的雌性群体均高于雄性群体,且随着生长发育和体质量差异的增大,雌、雄瞬时增长率的差异也呈增大趋势;雄性群体进入快速生长期的始速点和进入缓慢生长期的终速点均比雌性群体提前,雄性群体的快速生长期时间区间长度小于雌性群体,其对应的体质量区间长度也小于雌性群体,雌性群体在快速生长期的月增重显著高于雄性群体。研究表明,大菱鲆雌、雄群体在生长发育过程中存在着显著的生长差异,而形态差异不显著。     

大菱鲆促黄体激素受体基因的克隆及其生物信息学分析

通过兼并引物扩增及RACE技术,克隆大菱鲆促黄体激素受体(luteinizing hormone receptor,LHR)基因,分析其相应生物信息学特征和组织表达。结果显示:LHR基因序列全长3 184 bp,编码685个氨基酸,同大西洋庸鲽同源性最高。氨基酸序列分析发现,该基因编码蛋白为疏水性蛋白,相对分子量76.54 ku,理论等电点7.22,存在信号肽序列和跨膜区;亚细胞定位主要在内质网和细胞膜;预测该蛋白含有33个磷酸化位点、5个糖基化位点、6个富含亮氨酸重复序列、7个跨膜保守结构域,二级结构以无规则卷曲为主,三级结构其结构域呈α螺旋状构象;蛋白功能预测表明,LHR主要在辅助、运载、翻译和代谢调节过程中发挥关键作用。组织表达分析表明,LHR基因除在卵巢大量表达外,在其他非性腺组织也有表达,尤其是在肝脏表达较高。上述结果为深入探讨LHR基因在大菱鲆性腺发育过程中的生物学功能奠定了遗传信息基础,也为研究其他海水养殖鱼类生殖调控提供重要参考。     

皂角苷对大菱鲆非特异免疫的增强作用

为评价皂角苷增强鱼类非特异免疫效果,分别用浓度为0、15和35 mg/L的皂角苷浸泡大菱鲆,分析了浸泡后6、12、24、48和72 h时大菱鲆白细胞的吞噬活力、血清补体旁路途径溶血活性、血清与黏液中溶菌酶活性、抑菌活性与碱性磷酸酶(AKP)活性及血清总蛋白浓度。结果显示,皂角苷低浓度组(15 mg/L)与高浓度组(35 mg/L)大菱鲆白细胞吞噬活力分别在浸泡后12与24 h出现显著的增高(P0.05)。实验组大菱鲆血清中补体旁路途径溶血活性除低浓度组在12与24 h较对照组低外,其他检测时段均高于对照组(P0.05)。血清溶菌酶活性最高值出现在低浓度组浸泡后6 h时,黏液中未能检测出。不同时间点各组大菱鲆黏液抑菌活力变化幅度较大,两实验组黏液抑菌活力在浸泡后6与12 h时较对照组提高,两个实验组间无差异(P0.05)。血清中AKP活力高峰值出现在浸泡后6与12 h,该时段AKP的活力显著高于对照组,且实验组间存在差异(P0.05),而黏液中AKP高峰值出现在12 h,该时段AKP的活力显著高于对照组(P0.05),实验组间无差异。浸泡皂角苷后6 h高浓度组血清中蛋白浓度达到最高值,除12 h时各组差异不显著外(P0.05),24~72 h高浓度组中血清蛋白含量均高于低浓度及对照组(P0.05)。研究表明,皂角苷具有提高鱼类非特异免疫水平的效果。     

Pathology of Edwardsiella tarda infection in turbot, Scophthalmus maximus (L.)

Macroscopic and histopathological changes in cultured turbot, Scophthalmus maximus (L.), in Spain caused by infection with Edwardsiella tarda are described. Eye tumefaction, inflammation, haemorrhages, ascites and the presence of a purulent fluid were the main macroscopic lesions observed. Histopathological lesions were found in the kidney, spleen and liver. In the kidney and spleen these were characterized by a severe apostematous inflammatory reaction, with a large number of abscesses. The liver was affected to a lesser degree and only some phagocytes loaded with bacteria were observed. Ultrastructural observations indicated that macrophages were the main cell type implicated in the inflammatory response. Most of the bacteria observed within the phagocyte cytoplasm showed no degenerative changes and some were dividing. Degenerative changes observed in macrophages indicate their failure in preventing the infection.     

基于深度图像的大菱鲆体重估测模型研究

为了对大菱鲆体重进行无损估测,提出了一种基于深度图像的大菱鲆体重估测模型。该方法首先对大菱鲆深度图像进行图像预处理,提取出深度信息与生长数据进行映射建模,拟合出目标特征,并结合网格搜索优化支持向量回归(GS-SVR)算法,实现大菱鲆体重估测。估测结果与实际测量结果进行对比,两者的决定系数(R2)达到0.990 1,均方根误差(RMSE)为0.029 7。该模型具有简单灵活、估测精度高和易于实现等特点,同时具有很好的应用前景。     

n-3/n-6 HUFA对大菱鲆幼鱼生长性能、全鱼脂肪酸组成和血清生化指标的影响

为探讨饲料中不同n-3/n-6高不饱和脂肪酸(HUFA)对大菱鲆幼鱼生长性能、全鱼脂肪酸组成和血液生化指标的影响,配制了6种不同n-3/n-6 HUFA(D1:29.54,D2:23.04,D3:18.97,D4:9.06,D5:6.86,D6:3.87)的实验饲料。以大菱鲆幼鱼(12.18±0.01)g为研究对象,在循环水养殖系统中开展了为期8周的养殖实验。实验共分6组,每组3个重复,每个重复35尾鱼。结果显示:饲料中n-3/n-6 HUFA对大菱鲆幼鱼的成活率(SR)无显著影响;增重率(WGR)、特定生长率(SGR)和蛋白质效率(PER)呈先上升后下降趋势且D6组的显著低于其他各组,D2组蛋白质效率显著高于其他各组。全鱼粗蛋白和灰分均呈先上升后下降趋势;D6组肌肉粗蛋白和灰分显著低于其他各组。全鱼ARA含量随着n-3/n-6 HUFA的下降呈上升趋势;全鱼中EPA、DHA、n-3/n-6多不饱和脂肪酸(PUFA)和n-3/n-6 HUFA均随着饲料中n-3/n-6 HUFA的下降呈下降趋势。血清中酸性磷酸酶(ACP)和超氧化物歧化酶(SOD)随着n-3/n-6 HUFA的变化呈上升趋势;溶菌酶(LZM)和总抗氧化能力(T-AOC)呈先上升后下降的趋势,溶菌酶在D2组达到最大值,总抗氧化能力在D3组达到最大值。综上所述,饲料中n-3/n-6 HUFA在适宜范围(18.97~23.04)显著提高了实验鱼的生长性能和非特异性免疫力,改变了鱼体组织的常规成分和脂肪酸组成。