Changes in selected stress and immune‐related genes in Atlantic cod,Gadus morhua,following overcrowding

Using Atlantic cod, Gadus morhua, as a model for the stress response in gadoid fish, the changes in the expression of some stress and immune genes as well as the profiles of plasma cortisol were examined. Adult fish were kept at a density of ca. 100 kg m^?3 by lowering the water level in the rearing tank for 1 h and this short‐term crowding stress was repeated thrice over a 12‐h interval period. Blood samples were collected before exposure and at 2, 24 and 72 h post crowding. Plasma cortisol level significantly increased at 2 h post crowding but returned to pre‐crowding levels 24 h after exposure. The relative expression of the stress response genes, glucose transporter‐3 and a putative heat shock protein 70 significantly increased at 2 and 24 h post crowding respectively. Significant up‐regulation of the pro‐inflammatory cytokines, interleukin (IL)‐1β and IL‐8, as well as anti‐bacterial genes, g‐type lysozyme and bactericidal permeability‐increasing protein/lipopolysaccharide‐binding protein (BPI/LBP) was also observed at 2 h and the levels were maintained until 72 h post exposure, except for BPI/LBP which had maximum up‐regulation at 24 h. The present observations have implications with respect to fish welfare and assessment of the health status of the farmed fish.     

三角帆蚌外套膜表达的免疫相关基因筛选

利用三角帆蚌(Hyriopsis cumingii)外套膜cDNA文库EST序列,通过BLAST分析注释基因功能,发现35个与免疫防御功能相关的基因。根据其功能,这些免疫相关基因可划分为7类,即细胞免疫过程(2个)、蛋白酶和蛋白酶调节子(9个)、压力蛋白(7个)、抗菌肽(5个)、溶酶体酶(2个)、粘着蛋白(3个)及细胞凋亡和细胞周期调控相关基因(7个)。免疫相关基因在三角帆蚌外套膜中的广泛表达表明外套膜是重要的免疫组织。该研究为三角帆蚌外套膜分子免疫机理研究和抗病育种工作提供了基础性资料。     

Sodium alginate supplementation modulates gut microbiota,health parameters,growth performance and growth‐related gene expression in Malaysian Mahseer Tor tambroides

This study investigated the effects of sodium alginate supplementation on gut microbiota composition, health parameters, growth performances and growth‐related gene expression of Malaysian mahseer. Five test diets were formulated by supplementing 0%, 0.1%, 0.2%, 0.4% and 0.8% sodium alginate. Triplicate groups of juvenile Tor tambroides (2.19 ± 0.05 g) were stocked in 15 aquaria (20 individuals per aquarium) and fed at 3.0% body weight per day for 60 days. PCoA and UPGMA analysis showed that gut bacterial community were more convergence in higher sodium alginate‐supplemented diets. The percentage of Porphyromonadaceae, Bacteroides, Plesiomonas and Shewanella were substantially higher and Aeromonas, Entomoplasmatales and Prevotellaceae were drastically lower in higher sodium alginate (0.2%–0.8%) diets. Sodium alginate supplementation (≥0.2%) significantly improved the haematocrit value and respiratory burst activity of T. tambroides. Growth performances and feed utilization were significantly higher in 0.2%–0.4% sodium alginate‐supplemented diets. The increased growth rate of T. tambroides was governed by both hyperplastic and hypertrophic muscle growth. Real‐time PCR data demonstrated that most of the growth‐related genes were significantly upregulated in 0.2%–0.4% sodium alginate‐supplemented diet. Finally, it can be concluded that sodium alginate should be supplemented at 2 g/kg in practical fish feed formulation.     

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL‐8 and COX‐2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL‐1β, IL‐8 and COX‐2 and the antiviral factor like Mx genes performed with in vivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream and rock bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream were markedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)‐induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation‐related genes. The in vitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.     

虾源哈维氏弧菌的致病性与生物学特性比较分析

哈维氏弧菌(Vibrio harveyi)是导致养殖对虾暴发弧菌病的重要病原之一。从我国南方养殖凡纳滨对虾(Penaeus vannamei)分离、鉴定了10株哈维氏弧菌(Vh00947、Vh00949、Vh11011、Vh11014、Vh21217、Vh21218、Vh21220、Vh21229、Vh21231和Vh31487),分别肌注感染健康凡纳滨对虾后发现,菌株Vh21229致病性很弱,Vh00949其次,其它菌株毒力较强;对8种哈维氏弧菌常见毒力基因(Vh1、Vh2、Vh3、Vh4、tox S、hcp、zot和pap6)的检测显示,南方虾源哈维氏弧菌无论强弱毒株都很少携带zot、pap6、toxs和4种Vh基因(≤10%),表明这10个菌株的致病性与这7种常见毒力因子的相关度很低;hcp基因在所有菌株中均有检出,其中强毒株中检出率较高(50%),在较弱毒株(Vh00949)中也存在,表明hcp基因与这些菌株的致病性密切相关,但不是决定性因子。因此,这10株南方虾源哈维氏弧菌菌株的致病性差异应由这8种常见毒力因子以外的未知或未检测到的因子所决定。生化特征分析显示,所有菌株中只有弱毒株Vh21229不能利用D-甘露糖、蔗糖、D-海藻糖,而且只有其赖氨酸脱羧酶反应至阳性,而其它菌株均为阴性,推测导致哈维氏弧菌菌株生化特性改变的某种因子可能对菌株的致病性也产生了影响。药敏实验表明,10株哈维氏弧菌均对环丙沙星、氯霉素、恩诺沙星、美罗培南、头孢曲松、多西环素、头孢吡肟、诺氟沙星敏感,而对氨苄西林耐受性强,表明在虾类养殖过程中应当严格规范和控制抗菌药物尤其是青霉素类药物的使用。     

渔业数据失真对两种非平衡剩余产量模型评估结果的影响比较

为了研究渔业数据失真对两种非平衡剩余产量模型评估结果的影响,以南大西洋长鳍金枪鱼渔业产量和单位捕捞努力量渔获量(CPUE)数据作为基础数据,加入5种不同程度[变异系数(CV)=1%、5%、10%、20%和30%]的随机误差,模拟了(1)无数据失真,(2)仅产量数据失真,(3)仅CPUE数据失真,(4)产量和CPUE数据均失真等4种情况。利用基于ASPIC的非平衡剩余产量模型(ASM)和基于贝叶斯状态空间建模方法的非平衡剩余产量模型(BSM)分别评估了最大可持续产量(MSY)、B_(MSY)、F_(MSY)、B_(2011)/B_(MSY)、F2011/F_(MSY)等5种生物学参考点和管理指标。结果显示,在无数据失真情况下,ASM和BSM评估的MSY分别为2.866×10~4 t和2.836×10~4 t,B_(2011)/B_(MSY)分别为1.366和1.324,F2011/F_(MSY)分别为0.627和0.667,均相差不大,表明该渔业目前状态良好,ASM得到了较大的B_(MSY)(31.48×10~4 t)和较小的F_(MSY)(0.091);数据失真对ASM评估的B_(MSY)和F_(MSY)分别产生了严重的过低估计和过高估计,且CPUE数据失真产生的影响要比产量数据失真大;随着随机误差的增大,BSM评估的生物学参考点和管理指标的绝对百分比偏差有增大趋势;与ASM相比,BSM能够更好地处理渔业数据中存在的随机误差,除了MSY以外,BSM评估的生物学参考点和管理指标绝对百分比偏差均要比ASM的评估结果低,尤其是B_(MSY)和F_(MSY)。因此,在使用存在较大随机误差的渔业数据进行资源评估时,BSM具有一定的优势。     

长江中上游圆口铜鱼的种群死亡特征及其物种保护

根据葛洲坝(1998～2007年)、重庆(2006～2007年)和合江江段(1998～2005年)的渔业资源调查资料,对圆口铜鱼的种群死亡特征进行了评估,并利用Beverton-Holt模型的单位补充量渔获量方程,分析和探讨了长江中上游圆口铜鱼资源的合理利用.结果表明:1)3个江段各年间的圆口铜鱼开发率和捕捞死亡系数均远远大于相应年份最大允许的开发率和捕捞标准的基准尺度F0.1,3个江段的圆口铜鱼资源均处于严重过度捕捞状况;2)葛洲坝江段圆口铜鱼的最适开捕年龄为4龄(体长330mm),重庆和合江江段圆口铜鱼的开捕年龄应至少为5龄(体长375 mm).为保护圆口铜鱼的资源,建议葛洲坝江段三层流刺网的网日(2a)应大于75 mm为宜,重庆和合江江段的网目(2a)应不小于90 mm且同时需要控制各种渔具的日均作业次数.     

滨麦叶片转录组分析

滨麦（Leymus mollis）是小麦的野生近缘种,具有良好的耐盐抗旱和抗病虫害的能力,是小麦遗传育种的良好资源。为了深入了解和挖掘滨麦的基因信息和优异资源,本研究利用高通量测序技术对生长于滨海沙地的野生滨麦的叶片转录组进行了深度测序并组装,对得到的所有Unigene进行COG、GO和KEGG分类和功能注释,对Na⁺转运相关蛋白基因和氧化胁迫响应基因进行了挖掘,并通过实时荧光定量反转录PCR（qRT-PCR）对随机选取的12个基因的表达模式进行了验证。结果显示,转录组测序共获得112 846条Unigene,其中的59 380条得到功能注释,占总数的52.62%。COG分类结果表明,15 786条Unigene归属于25个类别。GO分类结果表明,20 350条Unigene被注释到三个大类中,其中,属于“生物学过程”的Unigene数量最多,占总数的43.56%。KEGG分析结果表明,18 550条Unigene得到注释,共涉及到128条代谢途径。其中,含Unigene数量最多的类别是“代谢通路”,涉及到的Unigene占总数的27.86%。“植物病原互作”和“植物激素信号转导途径”涉及的Unigene数量也较多,分别为1 367条和861条。对Na⁺转运相关蛋白基因和氧化胁迫响应基因进行挖掘,发现了15条注释为Na⁺/H⁺反向转运蛋白的Unigene和175条响应氧化胁迫的Unigene。随机选取的12个基因的qRT-PCR结果与转录组测序结果基本一致。     

富含多糖甜玉米幼穗RNA提取及高温胁迫基因表达分析

以改良Trizol试剂法从富含多糖的甜玉米幼穗中提取高质量、完整的总RNA,利用实时荧光定量PCR对高温胁迫下粤甜13雌穗发育的8个差异表达基因进行分析。结果表明,8个基因分为上调表达和下调表达,实时荧光定量PCR和测序法基因表达谱检测的基因上调或下调表达的趋势一致,上调表达基因ZM2G058057和下调表达基因ZM2G059964、ZM2G044670相对表达量较高,可能在高温胁迫影响甜玉米幼雌穗发育过程中起更重要的作用。