低渗诱导凡纳滨对虾肝胰腺消减文库的构建及其表达序列标签分析

初始体重(0.27±0.01)g的凡纳滨对虾(Litopenaeus vannamei)分别于盐度2和30中饲养56 d,采用抑制性消减杂交技术构建低渗胁迫诱导的凡纳滨对虾肝胰脏差异表达基因消减cDNA文库。消减杂交效率分析显示,正反向消减文库差异表达的基因各富集大约25和25~10倍。从正反向两个文库中随机挑选200个阳性克隆进行测序(正向文库80个,反向文库120个),共获得179条序列长度大于100 bp的ESTs序列,经CAP3在线拼接程序得到73条Unigenes(19条contigs和54条singlets)。生物学信息分析表明,这些Unigenes所代表的相应基因的功能包括免疫相关蛋白、代谢相关蛋白、核蛋白、细胞凋亡和转移蛋白。这些基因信息表明,凡纳滨对虾在低盐胁迫诱导所导致的生理变化非常复杂,构建的差异表达cDNA文库可较好地反映低盐胁迫对凡纳滨对虾影响的基因信息。     

Dietary guaiacol improves the growth of juvenile abalone, <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> Ino

We conducted a preliminarily study on the effects of dietary guaiacol on growth performance of abalone, Haliotis discus hannai Ino. Seven semi-purified diets were formulated with graded levels of guaiacol (0, 5, 10, 50, 100, 500, and 1 000 mg kg⁻¹). Abalone (initial weight: 0.29 ± 0.01 g; initial shell length: 8.55 ± 0.27 mm) were fed with these diets in a re-circulated water system for 152 days. Guaiacol significantly improved the specific growth rate (SGR) (P<0.05); excessive dietary guaiacol (1 000 mg kg⁻¹) led to significantly high mortality (P<0.05), and lipid content in the soft body increased significantly after dietary guaiacol (P<0.05). Activities of catalase and phenoloxidase (PO) in the viscera were significantly stimulated by dietary guaiacol (P<0.05). Broken-line analysis based on SGR indicated that the minimum dietary guaiacol for the optimal growth of juvenile abalone is 15.43 mg kg⁻¹.     

高蛋白棉粕替代鱼粉对凡纳滨对虾生长性能、非特异性免疫指标及抗病力的影响

【目的】研究高蛋白棉粕(High-protein cottonseed meal,HPCM)替代鱼粉对凡纳滨对虾幼虾生长性能、非特异性免疫酶活性及抗病力的影响。【方法】在基础饲料中分别添加质量分数0%、6.25%、9.375%、12.5%、18.75%的HPCM,分别替代饲料中质量分数0%、20%、30%、40%、50%和60%的鱼粉,配制6种等氮等能饲料,投喂初始体质量(0.38±0.01)g的健康凡纳滨对虾8周。【结果】HPCM替代鱼粉对凡纳滨对虾增重率、特定生长率、成活率、饲料系数及蛋白质效率影响均有统计学意义(P0.05),30%和40%替代组增重率和特定生长率最高,与对照组差异有统计学意义(P0.05),饲料系数最低,与对照组差异有统计学意义(P0.05),替代水平高于40%替代组时,蛋白质效率降低(P0.05);对凡纳滨对虾全虾体成分含量影响均有统计学意义(P0.05),随替代水平的增加,全虾中的水分、粗脂肪含量有升高趋势,粗蛋白含量呈先升后降趋势,粗灰分有降低趋势(P0.05);HPCM替代鱼粉对血清酚氧化酶(PO)、总超氧化物歧化酶(T-SOD)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)活性以及肝胰脏中丙二醛(MDA)含量影响有统计学意义(P0.05);攻毒试验表明,随着HPCM替代鱼粉比例及攻毒时间的增加,凡纳滨对虾存活率逐渐降低(P0.05)。【结论】以对虾增重率为判据,在鱼粉质量分数为25%的凡纳滨对虾基础饲料中HPCM替代鱼粉的适宜比例为37.44%。     

Effects of supplemental coated or crystalline methionine in low-fishmeal diet on the growth performance and body composition of juvenile cobia Rachycentron canadum （Linnaeus）

We evaluated the effects of supplemental coated and crystalline methionine （Met） on the growth performance and feed utilization of juvenile cobia （Rachycentron canadum Linnaeus） in a 60-d feeding trial. Fish groups were fed one of six isonitrogenous and isolipidic diets： 1） fishmeal control; 2） un- supplemented experimental （low-fish-meal diet deficient in Met）; or 3） one of four Met diets supplemented with crystalline L-Met, cellulose-acetate-phthalate coated L-Met, acrylic-resin coated L-Met, or tripalmitin- polyvinyl alcohol coated L-Met. The test diets were fed to triplicate groups of cobia （initial body weight 5.40-4-0.07 g） twice a day. The weight gain and specific growth rate of the fish fed the RES diet were highest among the Met-supplemented groups and were 23.64% and 7.99%, respectively, higher than those of the fish fed with the un-supplemented experimental diet （P〈0.05）. The protein efficiency ratio of the fish fed the MET diet was significantly higher than that of the fish fed the un-supplemented experimental diet and the fish in the other methionine supplementation groups （P〈0.05）. Our results suggest that supplementation of crystalline Met in low-fish-meal diets promotes the growth performance of juvenile cobia.     

低盐胁迫下凡纳滨对虾鳃丝抑制性消减杂交cDNA文库构建及差异ESTs分析

本研究以凡纳滨对虾(Litopenaeus vannamei)为对象,分别以低盐度胁迫下(4 ppt)凡纳滨对虾鳃丝cDNA为检测子,正常海水中凡纳滨对虾鳃丝cDNA为驱动予,采用抑制性消减杂交(suppression subtractive hybridization,SSH)技术构建了急性低盐胁迫诱导下凡纳滨对虾鳃丝消减cDNA文库。文库的重组率均高于95%,插入片段集中在250~750 bp之间,随机测序后在线拼接得到15组片段重叠群和37个单一序列共52个非重复序列。同源检索发现30个非重复序列与GenBank中的已知基因序列有较高的相似性。按差异表达基因编码的蛋白具体功能可分为6小类,依次为:(1)离子通道及转运蛋白;(2)蛋白合成翻译及转录因子;(3)应激抗氧化因子;(4)能量代谢;(5)信号受体和转导;(6)细胞纤维及骨架蛋白,所占比例分别为23.3%、20.0%、20.0%、16.7%、10.0%和10.0%。经功能聚类分析发现文库中含有大量离子通道蛋白及功能转运酶、胞内信号传导分子、细胞骨架蛋白等渗透调节和应激适应性相关基因,表明急性低盐胁迫诱导凡纳滨鳃丝抑制性消减杂交cDNA文库构建成功。     

谷胱甘肽酵母水解物对干露胁迫下凡纳滨对虾肝胰腺糖代谢相关基因的影响

【目的】探究富含谷胱甘肽的酵母水解物对干露胁迫下的凡纳滨对虾（Litopenaeus vannamei）肝胰腺糖代谢相关基因的影响,为谷胱甘肽酵母水解物在水产养殖中的应用提供参考。【方法】在基础饲料（鱼粉质量分数17.7%）中分别按质量分数0%、0.3%、0.5%、0.7%、0.9%、1.1%的比例添加谷胱甘肽酵母水解物,配制等氮等脂饲料6组（分别记为G0、G0.3、G0.5、G0.7、G0.9、G1.1）,饲喂初始体质量（0.24±0.01）g凡纳滨对虾4周,测定对虾生长性能指标;对对虾进行干露胁迫30 min,测定对虾肝胰腺相关基因的相对表达。【结果】各处理组对虾增重率、特定生长率和饲料系数无显著差异（P> 0.05）。干露胁迫后,G0组凡纳滨对虾肝胰腺缺氧诱导因子（HIF-1α）、果糖磷酸激酶（PFK）和乳酸脱氢酶（LDH）基因的表达显著上调（P <0.05）,琥珀酸脱氢酶（SDH）、苹果酸脱氢酶（MDH）和细胞色素氧化酶（CCO）基因显著下调（P <0.05）,葡萄糖转运蛋白1(GLUT1)、己糖激酶（HK）和丙酮酸激酶（PK）基因的表达无显著变化（P>...