Imbalanced lysine to arginine ratios reduced performance in juvenile cobia (Rachycentron canadum) fed high plant protein diets

Cobia (8.4 ± 0.1 g body weight) were fed to satiation with three test diets of high plant protein‐based ingredients and different lysine to arginine ratios, and one commercial diet (currently used for cobia rearing in Vietnam as a control for growth) for 6 weeks. The test diets contained 206 g marine ingredients kg^?1, including fishmeal, krill meal and fish protein concentrate (in order of high to low inclusion), while the rest of the dietary protein was a blend of soya and pea protein concentrate, wheat protein and sunflower meal. Crystalline lysine and arginine were added in the test diets to produce either a balanced lysine to arginine ratio (BL/A; 1.1) and a high or low lysine to arginine ratio (HL/A; 1.8 and LL/A; 0.8, respectively). There were no significant differences in final body weight, weight gain, feed conversion ratio or protein gain between cobia fed BL/A‐ and commercial control diet (CCT). Cobia fed BL/A diet performed better than fish fed either HL/A‐ or LL/A diet. This was partly due to a higher feed intake and protein and lipid gain in cobia fed BL/A diet as compared to HL/A‐ and LL/A diet.     

Effects of different arginine/lysine level on growth performance,body composition and digestive enzyme activity of Macrobrachium rosenbergii

Given the universal presence of antagonism between arginine and lysine, determining the optimal dietary arginine/lysine level is of great importance. In this study, eight diets were formulated to contain different arginine/lysine level, followed by a 9‐week feeding trial with shrimps of 2.10 ± 0.01 cm in length and 0.38 ± 0.02 g in weight to determine the effect of dietary arginine/lysine level on growth performance, body composition and enzyme activity. The results showed that the Diet 6 (arginine/lysine level of 29.4/21.0 g/kg) group had significantly higher weight gain rate, and specific growth rate than the control group and most of the other groups (p < .05) did. Similarly, the retention efficiency of arginine and lysine in the Diet 6 group was significantly higher than the control group and most of the other groups (p < .05). Moreover, most digestive enzymes had higher activity levels in the Diet 6 group, and evident antagonism between dietary arginine and lysine was detected. In summary, dietary arginine/lysine level of 29.4/21.0 g/kg produced the best growth performance, suggesting that a better growth performance in Macrobrachium rosenbergii culture can be obtained by adjusting the dietary arginine/lysine level.     

锯缘青蟹精氨酸激酶基因的克隆与表达

通过分子生物学方法,克隆得到锯缘青蟹（Scylla serrata）精氨酸激酶（Arginine Kinase,AK）开放阅读框基因序列.测序结果显示：该开放阅读框基因的序列长度为1 071 bp,编码356个氨基酸残基;该序列登录Genbank（GQ：851626）,序列比对结果显示,锯缘青蟹精氨酸激酶与凡纳滨对虾（Litopenaeus vannamei）精氨酸激酶的氨基酸序列同源性高达94%,与岸蟹（Carcinus maenas）、中华绒鳌蟹（Eriocheir sinensis）等甲壳类动物的精氨酸激酶也有较高的同源性,均高于90%;将克隆得到的锯缘青蟹精氨酸激酶基因与pGEX-4T-3载体连接,转化大肠杆菌BL21,经IPTG诱导得到分子质量为65 ku的融合表达蛋白（GST-AK）,经兔抗锯缘青蟹精氨酸激酶多克隆抗体的免疫印迹分析证实,GST-AK具有抗原性.     

氧化鱼油对黄颡鱼生长性能和抗氧化指标的影响及精氨酸的干预作用

本试验旨在研究饲料中添加氧化鱼油对黄颡鱼(Pelteobagrus fulvidraco)生长性能、体成分、血清生化指标和血清、肝脏抗氧化指标的影响以及添加精氨酸对其的干预作用。选取初始体重为(4.41±0.05)g的健康黄颡鱼幼鱼600尾,随机分为6组,每组4个重复,分别投喂含2.5%新鲜鱼油(FF组)、1.5%新鲜鱼油+1.0%氧化鱼油(FO1组)、0.5%新鲜鱼油+2.0%氧化鱼油(FO2组)、2.5%新鲜鱼油+0.48%L-精氨酸盐酸盐(FFA组)、1.5%新鲜鱼油+1.0%氧化鱼油+0.48%L-精氨酸盐酸盐(FOA1组)、0.5%新鲜鱼油+2.0%氧化鱼油+0.48%L-精氨酸盐酸盐(FOA2组)的6种饲料,投喂期为56 d。结果显示:在FF、FO1、FO2组中,随着氧化鱼油添加量的增加,黄颡鱼的增重率、特定生长率和蛋白质沉积率均逐渐降低,饲料系数和摄食率均逐渐升高,均在FO2组达到极值,与其他2组差异达到显著水平(P0.05);氧化鱼油饲料添加精氨酸后,在FFA、FOA1、FOA2组上述指标没有产生显著性差异(P0.05),其中FOA1、FOA2组的增重率分别比FO1、FO2组升高3.0%和9.9%。双因素方差分析结果显示,氧化鱼油对黄颡鱼增重率和摄食率的影响达到显著水平(P0.05),氧化鱼油与精氨酸对黄颡鱼特定生长率、蛋白质沉积率和饲料系数的影响存在交互作用(P0.05)。FO1组肝体比显著低于FF组(P0.05),FO2组肠体比与FFA、FOA2组相比显著降低(P0.05),氧化鱼油与精氨酸对黄颡鱼肝体比的影响存在交互作用(P0.05)。FOA1组全鱼粗脂肪含量与FO1组相比显著降低(P0.05)。在FF、FO1、FO2组中,随着氧化鱼油添加量的增加,黄颡鱼血清总抗氧化能力(T-AOC)逐渐降低,FO2组显著低于FF组(P0.05);氧化鱼油饲料添加精氨酸后,黄颡鱼血清T-AOC分别升高77.0%(FOA1组vs.FO1组)和137.4%(FOA2组vs.FO2组),其中后者达显著水平(P0.05)。双因素方差分析结果显示,氧化鱼油对血清丙二醛(MDA)含量的影响达到显著水平(P0.05),氧化鱼油和精氨酸对黄颡鱼血清T-AOC的影响存在交互作用(P0.05)。结果表明,在饲料中添加一定量的氧化鱼油会抑制黄颡鱼的生长并降低血清的抗氧化能力,但添加一定量的精氨酸可以缓解氧化鱼油对黄颡鱼生长的抑制作用,并增强其机体的抗氧化能力。     

菜薹蛋白精氨酸甲基转移酶基因BrcuPRMT5的克隆及表达分析

以‘油青49’和‘油青甜菜薹80’菜薹茎尖为材料,采用RT-PCR和RACE技术克隆获得组蛋白甲基转移酶基因Brcu PRMT5的全长cDNA和gDNA序列。BrcuPRMT5 cDNA序列全长为2 117 bp,其中完整开放阅读框为1 929 bp,编码642个氨基酸,相对分子量为71.55 kD,理论等电点(p I)为5.87;多序列比对结果表明,Brcu PRMT5编码的氨基酸序列含有高等植物PRMT5基因1个高度保守的结构域;系统发育分析结果显示与大白菜、油菜及甘蓝的亲缘关系最近;亚细胞定位软件分析得知,Brcu PRMT5蛋白无跨膜区域,可能定位于线粒体中;对应的gDNA全长为4 151 bp,含有23个外显子和22个内含子,最长的外显子长度为140 bp,内含子的长度范围为50~150 bp。利用半定量RT-PCR和实时荧光定量PCR技术分析基因的表达,BrcuPRMT5在菜薹不同组织中均有表达,其中在花中表达量最高,叶次之,根中最低;BrcuPRMT5从苗期至完全抽薹开花期的表达量呈现上升趋势。BrcuPRMT5在菜薹花发育过程中可能起一定的调控作用。     

粘虫EF-1a和AK基因的克隆及real time RT-PCR方法建立

为今后利用SYBR GreenⅠ荧光定量法研究粘虫EF-1a和AK基因的表达情况，设计合成用于PCR扩增的引物，运用RT-PCR方法克隆EF-1a和AK基因片段，并进行序列同源性分析；然后采用SYBR Green Ⅰ荧光定量法设计特异性引物进行RT-PCR扩增，通过熔解曲线和标准曲线分析等建立实时RT-PCR方法。结果表明，从粘虫中克隆获得EF-1a和AK基因片段长度分别为622 bp和1020 bp，分别编码207个氨基酸和339个氨基酸；序列同源性分析结果表明，粘虫EF-1a与鳞翅目昆虫家蚕、桦尺蠖和柑桔凤蝶的EF-1a氨基酸序列同源性为95%以上；AK与鳞翅目昆虫草地贪夜蛾、烟蚜夜蛾、棉铃虫、斜纹夜蛾和甜菜夜蛾的AK氨基酸序列同源性为98%以上。熔解曲线和标准曲线结果显示，设计的EF-1a和AK基因引物均可应用于实时RT-PCR扩增。     

颈静脉灌注精氨酸对泌乳中期奶牛血清生化和免疫指标的影响

本文旨在研究颈静脉灌注精氨酸对泌乳中期奶牛血清生化和免疫指标的影响。采用3×3复式拉丁方试验设计,选择6头体重、胎次、泌乳期、泌乳量和体况基本一致的荷斯坦奶牛为试验动物,随机分为3组(每组2头),分别为酪蛋白组(对照组)、精氨酸组(灌注37.66 g/d)、丙氨酸组(与精氨酸组等氮,灌注77.24 g/d);每个试验期为22 d(7 d灌注+15 d间隔),在每个试验灌注期的最后1 d晨饲前采血,测定血清生化和免疫指标。结果表明:精氨酸组的奶牛血清中总蛋白、免疫球蛋白G(Ig G)、肿瘤坏死因子α(TNF-α)浓度要显著高于其他2组(P0.05),白球比、白细胞介素1(IL-1)浓度显著高于丙氨酸组(P0.05),但是牛奶中体细胞数显著低于其他2组(P0.05),而血清中谷丙转氨酶、谷草转氨酶活性,免疫球蛋白A(Ig A)、免疫球蛋白E(Ig E)、白细胞介素6(IL-6)浓度在组间无显著差异(P0.05)。综上,灌注精氨酸能够通过提高血清总蛋白、Ig G及IL-1、TNF-α的浓度,从而一定程度上增强泌乳奶牛机体的免疫力和胎儿的被动免疫力。