Essential amino acids supplementation to practical diets affects growth,feed utilization and glucose metabolism‐related signalling molecules of blunt snout bream (Megalobrama amblycephala)

Six diets were formulated to investigate the success of fish meal (FM) replacement by plant proteins; diet 1 reflected a commercial feed (8% FM), diet 3 contained 4% FM, and diet 5 was devoid of FM. Whereas, diet 2, diet 4, and diet 6 reflected diet 1, diet 3, diet 5, respectively, and supplemented with essential amino acid (EAA). At the end of 8‐week trial, there was no significant difference in survival rate. Significantly higher final weight, weight gain rate, and specific growth rate were recorded in the group fed diet 2 compared with the other treatments (except diet 4) (p < 0.05). Feed conversion ratio of fish fed diet 2 was significantly lower than those fed diets 1, 3, and 5 (p < 0.05). The lowest feed intake and highest protein efficiency rate were found in fish fed diet 2 (p < 0.05). There was no significant difference in whole body compositions between treatments. Plasma aspartate transaminases, alanine aminotransferase, and glucose were significantly affected by dietary treatments (p < 0.05), while plasma protein and albumin contents were not influenced by the treatments. The relative expression of target of rapamycin (TOR) and phosphatidylinositol 3‐kinase, regulatory subunit 1 (alpha) (PIK3R1) in fish fed diet 3 (4% FM) were significantly down‐regulated compared with those fed diet 6 for TOR and diets 4 and 6 for PIK3R1 (p < 0.05). Insulin receptor substrate 1 (IRS‐1) and janus kinase 3 (JAK3) expressions were fluctuated, with the higher levels in fish fed diets 4 and 6. In conclusion, the findings of this study indicate that plant protein mixture supplemented with EAA could be used to substitute FM in practical diet for Megalobrama amblycephala.     

团头鲂tf和tfr1a基因启动子克隆及转录调控分析

为探索鱼类转铁蛋白基因tf和转铁蛋白受体基因tfr1a的转录调控机制,本实验以团头鲂为研究对象,在其全基因组数据库中获取tf和tfr1a基因序列,对2个基因候选启动子区转录因子结合位点及CpG岛进行预测,通过PCR方法克隆得到tf和tfr1a基因近端启动子区不同长度片段,连接至pGL3-Basic/pEGFP-1载体,瞬时转染入Hela细胞,并采用双荧光素酶报告基因检测系统进行检测。结果发现,团头鲂tf基因启动子区无CpG岛位点,而tfr1a基因启动子区有2个CpG岛位点。成功构建9个tf和10个tfr1a不同长度启动子片段的重组质粒,经双荧光素酶报告基因系统检测发现,tf启动子核心区域为-268^+56 bp,且-1 308^-1 102 bp片段可能存在正调控该基因表达的转录因子结合位点;tfr1a启动子核心区域为-224^+48 bp,且+48^+92 bp可能存在抑制该基因转录的负调控元件,而-1 229^-1 219 bp区域可能存在促进tfr1a基因表达的正调控转录因子结合位点。     

Growth and ontogenetic development of digestive functionality in black Amur bream (Megalobrama terminalis)

The function of digestive physiology during ontogenetic development is essential to ensure high survival and growth rates. In order to evaluate the digestive physiological capacity of the black Amur bream (Megalobrama terminalis), changes of morphology and digestive enzyme activity (trypsin, lipase, amylase, pepsin, leucine aminopeptidase and alkaline phosphatase) in larvae were examined from hatching to 40 days after hatching (DAH). Results indicated that fluctuation patterns differed between the total and specific activities of the digestive enzymes. The total activities of these six enzymes gradually increased throughout the fish growth. The specific activity of trypsin peaked at 5 DAH and then decreased dramatically, while it increased remarkably again from 8 to 10 DAH and remained stable level after 20 DAH. Pepsin activity was first examined in M. terminalis at 15 DAH and gradually elevated towards the end of the experiment. The specific activity of lipase displayed obvious peaks at 5 and 20 DAH. For the amylase, its specific activity reached plateau at 4 DAH, underwent sharp decrease, and remained stable after 20 DAH. In addition, we found that the specific activity of alkaline phosphatase raised significantly from hatching to 5 DAH, and tended to keep slight fluctuation after 15 DAH. From the above, we concluded that the specific activities of digestive enzymes in the larvae varied constantly from 3 to 20 DAH and turned relatively stable after 20 DAH. The present study provides effective information that is useful to improve the seedling cultivation and the technology for healthy breeding.     

厚颌鲂外周血细胞形态及细胞化学特征研究

采用血细胞常规染色和细胞化学染色对厚颌鲂(Megalobrama pellegrini)外周血细胞形态及细胞化学特征进行观察。结果显示:厚颌鲂外周血细胞可分为红细胞、血栓细胞、单核细胞、淋巴细胞和三种类型粒细胞。红细胞有幼稚、成熟和衰老三种形态,其中幼稚和衰老红细胞较少见;血栓细胞数量最多,其次是淋巴细胞。粒细胞中Ⅰ型和Ⅱ型较常见,Ⅲ型粒细胞少有。细胞化学染色显示幼红细胞、血栓细胞和淋巴细胞均呈ACP弱阳性;单核细胞呈ACP和ANAE阳性;Ⅰ型粒细胞是嗜中性粒细胞,POX、ACP和ANAE阳性;Ⅱ型粒细胞是PAS-GL,PAS强阳性和ACP阳性;Ⅲ型粒细胞是Ⅰ型粒细胞的晚期形态。厚颌鲂的PAS-GL可能是嗜酸性/嗜碱性粒细胞,而不是嗜碱性粒细胞的前体。     

团头鲂肠道肝胰脏消化酶的初步研究

本文采用动力学方法测定了团头鲂肝胰脏淀粉酶和肠道蛋白酶、淀粉酶的活性分布及pH值对各消化酶活性的影响,结果表明:食糜中的蛋白酶、淀粉酶活性沿消化道从前至后逐渐减弱,而肠黏膜中各消化酶活性则基本上是由前到后逐渐增加。在pH值6.0～9.0范围内,肠道蛋白酶活性在pH为8.6时为最大,在pH值6.2～8.6范围内,肝胰脏和肠道淀粉酶活性在pH值7.8时为最大。     

Induction of interspecific allo-tetraploids of Megalobrama amblycephala♀×Megalobrama terminalis♂ by heat shock

Interspecific allo-tetraploidy was induced by a combination of hybridization of Megalobrama amblycephala ♀× Megalobrama terminalis ♂ and thermal shock. A thermal shock at 40 °C for 2 min applied to eggs 33 or 26 min (τ₀: 1.40) after fertilization resulted in a significantly ( P <0.01) higher yield of allo-tetraploids than hybrid crosses without heat shock. The yield of allo-tetraploids in embryos from mid-gastrula to hatching stages was 15.7–24.4% in different thermal shock treatment groups. The allo-tetraploidy rate in 330-day-old juveniles was 9.3% and 6.7% in different year treatment groups. After 2 years of rearing, allo-tetraploidy males attained sexual maturity with milk-like milt at the age of 2+, while allo-tetraploidy females at the age of 2+, and even at 3+, did not show any signs of sexual maturation exhibiting obviously delayed maturity. Only a few allo-tetraploidy females (three individuals) showed maturation characters at the age of 4+ and these females were induced to spawn.     

厚颌鲂的年龄结构及生长特性

以鳞片作为鉴定年龄的材料,对龙溪河厚颌鲂(Megalobrama pellegrini)种群开展年龄结构与生长特性的研究。结果表明,厚颌鲂鳞片年轮结构呈疏密切割型,年轮特征显著,可用于年龄鉴定,3~4月是年轮形成高峰时间。种群由0~7龄共8个龄组组成,以低龄个体为主,1~3龄个体占83.14%,体长分布主要集中在130~250 mm间(77.71%)。体质量分布主要集中在50~250 g(64.33%)。群体总性比为♀∶♂=1∶1.03,符合1∶1理论比值。体长和鳞径呈直线关系,体长和体质量呈幂函数关系且幂指数接近3,属于匀速生长类型,因此用Von Bertalanffy方程描述其生长特性,相关生长参数分别为:渐近体长L∞=488.89 mm,渐近体质量W∞=2486.96 g,k=0.1648,t0=-0.5453龄。体质量生长拐点Ti=6.40龄,对应体长、体质量分别为Lt=333.14 mm,Wt=746.06 g。厚颌鲂的总体生长性能逊于同属其他鱼类。     

团头鲂细菌性败血症免疫研究

将温和气单胞菌(Aeromonas sobria)复壮，在28℃条件下加福尔马林灭活48h，制备成灭活苗。用该疫苗免疫团头鲂，取免疫后第10d，20d，30d的血清做凝集抗体试验，最高凝集抗体效价达1：32～128，对照组为1：4～8；免疫鱼的抗感染保护力达70％。     

三角鲂与团头鲂正反杂交F1的遗传性状

采用聚类分析、主在分分析和判别分析3种数理统计方法，对团头舫、三角鲂及其正反杂交F1的比例性状和框架参数进行分析，探讨了亲本形态性状在子代中的遗传传递情况。结果表明：（1）正反交F1形态都表现出较多的母性遗传特征，但三角鲂母本对杂交F1遗传特征的影响强于团头鲂母本。（2）躯干部特征、头尾轴特征及背棘等为区分团头鲂、三角鲂及其正反杂交F1的重要因子。     

广东鲂(♀)×团头鲂()杂交子一代及其双亲同工酶的比较

采用聚丙烯酰胺垂直平板电泳法对广东鲂、团头鲂及其杂交子一代肌肝眼 3种组织的 4种同工酶(EST、LDH、MDH、SOD)进行电泳 ,分析其酶谱组成和活性差异。结果显示父母本同种组织中大部分同工酶的表达酶谱较相似 ,尤其以眼组织的LDH、肝组织的MDH以及 3种组织的SOD同工酶的表型最为相似 ,表明两种鱼的亲缘关系较近 ,是本远缘杂交成功的遗传物质基础。同时父母本的某些同工酶谱又存在有稳定的种间差异 ,如肝组织的LDH。杂交子一代同工酶表型有些具有来自双亲的区带总和 ,有些则单独表达父本或母本的基因 ,更有些出现父母本没有的表型