低浓度Cd2+胁迫对青鱼组织SOD活性和MT诱导的影响

采用静水试验方法,研究了Cd2+对青鱼的急性毒性效应以及Cd2+安全浓度胁迫对青鱼鳃丝、肌肉和肝脏的超氧化物歧化酶(SOD)和金属硫蛋白(MT)的影响。结果表明:Cd2+对青鱼的96 h LC50为1.434 mg/L,Cd2+对青鱼的生态毒性为高毒。在0.143 mg/L(安全浓度)Cd2+胁迫下,各组织中SOD活性和MT含量在6 h、12 h内显著上升,呈现出一定时间-效应关系,以暴露6 h时最为显著(P0.05);暴露12 h后,MT的含量基本上趋于平衡,而SOD活性却是快速下降,表明了12 h后该浓度Cd2+污染胁迫对SOD活性的抑制作用。整个试验期间,对照组青鱼各组织中的SOD活性和MT含量大小顺序皆为肝脏鳃丝肌肉,同种组织内相对比较稳定;肌肉中的SOD活性要显著低于鳃丝和肝脏,而肝脏中的MT含量要显著高于肌肉和鳃丝。     

日本沼虾半胱氨酸蛋白酶抑制因子基因的克隆及其组织表达特征

半胱氨酸蛋白酶抑制因子(CST)是一种能够可逆结合并抑制半胱氨酸蛋白酶类的活性的蛋白,为了明确日本沼虾(Macrobrachium nipponense)CST基因(Mn CST)序列及其在日本沼虾卵巢中的作用,本研究利用RACE方法克隆获得日本沼虾Mn CST全长cDNA序列,利用qRT-PCR分析了其在不同组织和不同发育阶段的卵巢中的表达情况,并利用RNA干扰(RNAi)技术初步分析了Mn CST在日本沼虾卵巢发育中的作用。结果显示,Mn CST cDNA序列全长6199 bp,包含1183 bp 5′非编码区、2304 bp 3′非编码区和2709 bp开放阅读框,编码903个氨基酸残基,其氨基酸序列上有6个cystatin-like(半胱氨酸蛋白酶抑制因子样)结构域和42个磷酸化位点;Mn CST基因在各个组织中均有表达,肠中表达量最高;日本沼虾不同发育阶段的卵巢中均有Mn CST基因表达,Ⅳ期卵巢中表达量最高,Ⅱ期卵巢中表达量最低;Mn CST基因在日本沼虾卵巢中的表达变化与组织蛋白酶B(Cts B)和组织蛋白酶L(Cts L)基因的表达变化基本是一致的,但对卵黄蛋白原(Vg)基因的表达没有直接影响。Mn CST在日本沼虾卵巢中的作用可能主要是抑制Cts B和Cts L的活性,在日本沼虾卵巢发育过程中对Vg的水解起间接的调控作用。     

Replacement of Fishmeal with a Blend of Poultry Byproduct Meal and Soybean Meal in Diets for Largemouth Bass,Micropterus salmoides

A feeding trial was conducted to evaluate the potential of replacing fishmeal with poultry byproduct meal (PBM) and soybean meal in diets for largemouth bass, Micropterus salmoides. A reference diet (C) contained 400 g/kg fishmeal, and 40 or 60% of the fishmeal was replaced with a blend of pet‐food‐grade PBM and soybean meal (diets PP1 and PP2) or a blend of feed‐grade PBM and soybean meal (diets PF1 and PF2). No significant differences were found in weight gain, nitrogen retention efficiency (NRE), condition factor, and body composition among fish fed diets PP1, PP2, PF1, and PF2. Feed intake and feed conversion ratio (FCR) were higher in fish fed diet PF1 than in fish fed diet PP1. No significant differences were found in weight gain, NRE, condition factor, and body composition between fish fed diet C and diets PP1, PP2, PF1, and PF2. The feed intake and FCR were lower in fish fed diet C than in fish fed diets PP2, PF1, and PF2. This study reveals that dietary fishmeal level for largemouth bass could be reduced to 160 g/kg by inclusion of PBM and soybean meal in combination.     

Genetic diversity and population structure of a giant freshwater prawn (Macrobrachium rosenbergii) breeding nucleus in China

The giant freshwater prawn breeding programme in China has been carried out as a closed nucleus from 2006. The programme aimed to improve the harvest body weight and pond survival and has achieved remarkable progress. In this study, the genetic diversity and population structure of the giant freshwater prawn breeding nucleus was analysed. Pedigree completeness, the status of genetic diversity, inbreeding accumulation and effective population size of the breeding nucleus were analysed according to the pedigree record. From 2006 to 2014, a total of 107,941 individuals with 555 males and 967 females in nine generations were included in the analyses. Pedigree completeness index in six generations was at least 0.92. After performing eight selections, the genetic diversity decreased by 3%, 67% of which was caused by random genetic drift. After performing 2 years’ selection, the inbreeding level began to show a marked upward trend. The regression of the average rate of inbreeding and the average rate of co‐ancestry was 0.0041 and 0.0051 from 2006 to 2014. The effective population size calculated from the above two parameters was 122 and 98 respectively. The effective population size is still within the recommended level for a population to continue the genetic improvement programme, but below the level required (500) for retaining the evolutionary potential. To restrain the increasing level of inbreeding and maintain effective population size, measures are required to develop a breeding programme which integrates management of genetic variability and selection.     

牛蛙源无乳链球菌全菌疫苗免疫效果研究

将分离鉴定的无乳链球菌用0.4%的甲醛灭活,加弗氏完全佐剂制成灭活全菌苗,对牛蛙进行注射免疫。通过测定受免牛蛙血清中凝集抗体效价,总蛋白、免疫球蛋白的测定及免疫保护率的测定,探讨疫苗的免疫应答状况。研究结果表明,无乳链球菌全菌疫苗能够提高牛蛙的血清抗体水平及总蛋白、免疫球蛋白含量。此外,无乳链球菌全菌苗对牛蛙抗人工感染也具有较好的免疫保护作用。因此,将无乳链球菌全菌苗用于牛蛙链球菌病的防治具有广阔的发展前景。     

一株亚硝酸盐降解菌的分离鉴定

为寻找对亚硝酸盐具有高效降解能力、稳定和安全的优良菌株,采用亚硝酸盐氧化细菌(Nitrite-oxidizing bacteria, NOB)富集、分离技术从湖州某水域淤泥中分离得到一株纯培养菌株N3。提取细菌的总DNA,利用细菌16S rDNA特异性引物进行多聚酶链反应(PCR)扩增;扩增产物经琼脂糖凝胶电泳和Sanger末端终止法测序分析,用NCBI-Blast软件将测序结果在Genbank等数据库中进行同源性检索。结果表明,N3的DNA扩增产物片断大小为1441 bp,测序结果经检索证实N3与标准菌株Cellulosimicrobium cellulans (AY665978.1) 16S rDNA保守性片段有100%的同源性,可以判定所分离得到的N3菌株为纤维单胞菌属的纤维化纤维单胞菌(Cellulosimicrobium cellulans)。     

正圆形珠核规格与育珠蚌体长的匹配试验

报导了在三角帆蚌内脏团植入正圆形珠核与蚌体长之间的匹配实验结果。比较了不同规格的蚌体对有核珍珠留核率、成珠率、尾巴珠率、光洁度、光泽度、正圆度及珍珠等级的影响。实验结果表明,对于8.00～8.25 mm的珠核而言,植入12.0～12.2 cm的蚌体的内脏团效果比较理想。     

嗜水气单胞菌的研究进展

简要回顾和总结了气单胞菌的分布、分类地位、分离鉴定和生长条件等方面的研究情况,并概述嗜水气单胞菌的外膜蛋白（OMP）、胞外分泌物（ECP）等致病因子、诊断技术及疫苗开发方面新的研究成果。     

哈维氏弧菌的主要致病因子及其特性分析

从大黄鱼致病菌哈维氏弧菌(Vibrio harveyi)GYC1108-1株提取外膜蛋白(OMP)、脂多糖(LPS)和胞外产物(ECP),并进行毒性实验,以研究哈维氏弧菌的主要致病因子及其特征.结果表明:OMP、LPS致病性较弱,而ECP可导致鱼死亡,初步得出哈维氏弧菌GYC1108-1株的主要致病因子为胞外产物,其对鱼的半数致死量(LD50)为3.5 μg·g-1;该蛋白酶与底物偶氮酪蛋白(azocasein)作用的最适pH为8.0,最适温度28℃,对热敏感,分子质量为55 ku;该酶活性被碘乙酸抑制,也被SDS和HgCl2完全抑制,PMSF和ZnCl2能部分抑制该蛋白酶活性,而CuCl2、CaCl2、MgCl2对该酶活性影响不大,2-巯基乙醇、DTT、L-半胱氨酸、EDTA和EGTA对该酶活性有促进作用,表明其为半胱氨酸蛋白酶;研究发现ECP具有酪蛋白酶、明胶蛋白酶、淀粉酶、卵磷脂酶和脂肪酶活性,无脲酶活性;免疫印迹结果发现GYC1108-1的ECP能与大黄鱼抗GYC1108-1血清发生免疫发应,分子质量为55 ku的半胱氨酸蛋白酶是具有免疫原性的物质.     

五倍子、乌梅等45种中草药提取物对锯缘青蟹致病弧菌的抑菌作用研究

取五倍子、乌梅等45种中草药,分别采用水提取法和醇提取法制备提取液,除菌后滴加至中6mm空白药敏滤纸片上,制成试验用药敏纸片。测试了提取物对拟态弧菌SM060818、梅氏弧菌SM060813、霍利斯弧菌SM060913等3种分离自锯缘青蟹体外抑菌的效果。结果表明：所试验的45种中药提取物中,有28种中药提取物对试验弧菌表现有抑菌作用;其中五倍子、乌梅、秦皮、艾叶中药提取物抑菌作用较好,对3种菌株均有较好抑菌效果;天冬、白头翁、赤芍、金银花、黄精、黄柏、青蒿等11种中草药提取物仅对2种菌株有较好抑制作用,千里光、穿心莲等17种中药提取物对弧菌无抑制作用;大部分中药抑菌活性成份均为水溶性提取物,青蒿存在明显的醇溶性抑菌物质。测定了五倍子等9种中药提取物对致病弧菌的最低抑菌浓度（MIC）,五倍子、乌梅和天冬3种药物对拟态弧菌和梅氏弧菌的MIC分别为0．172mg／mL、0．461mg／mL、1．042mg／mL,对霍利斯弧菌的MIC分别为0．086mg／mL、0．922mg／mL和1．042mg／mL,有较好的抑菌作用。