氯虫苯甲酰胺和氟虫腈对黏虫细胞色素P450基因表达的影响

为筛选出黏虫Mythimna separata参与杀虫剂解毒代谢的主效细胞色素P450基因,采用叶片浸渍法测定了用于处理黏虫3龄幼虫的亚致死浓度,通过构建转录组测序文库并结合数字基因表达(digital gene expression,DGE)对不同处理的黏虫进行测序,并运用实时荧光定量PCR(realtime quantitative polymerase chain reaction,RT-qPCR)技术验证12个P450基因的表达情况。结果表明,用于处理黏虫的氯虫苯甲酰胺和氟虫腈亚致死浓度LC_(10)分别为0.15、13.66 mg/L;对照样品、氟虫腈处理样品和氯虫苯甲酰胺处理样品分别获得59 521 504、64 838 148和41 722 990个原始序列数据,分别获得57 441 216、62 368 912和40 285 164个过滤后的序列数据;过滤后的序列长度分别为8.62、9.36和6.04 G;碱基错误率均为0.02%;Phred数值大于20、30的碱基占总碱基的百分比均高于90.59%;鸟嘌呤+胞嘧啶(guanine cytosine,GC)含量分别为47.16%、48.94%和47.55%,表明转录组测序质量较高;黏虫受氯虫苯甲酰胺胁迫后,29个P450基因表达量上调,27个P450基因表达量下调;黏虫受氟虫腈胁迫后,23个P450基因表达量上调,26个P450基因表达量下调;12个P450基因表达量的RT-qPCR技术检测结果与DGE测序文库显示的结果基本一致。     

渔业管理中生物学参考点的理论及其应用

生物学参考点常被表示为与渔业管理相关联的捕捞死亡率和生物量,是单从生物学角度来衡量渔业资源及其开发状况的指标。通常可分为目标参考点、限制参考点和阈值参考点。目标参考点是为了持续获得某一目标渔获量所需的最小生物量和相应捕捞死亡率,包括目标生物量(Bmsy)、目标产卵亲体量(Smsy、SSB35%、SSB40%)、目标捕捞死亡率(Fmsy、Fmax、F0.1、Fmed、F40%、F40%)等参数。限制参考点用于保证捕捞死亡率不会高到危害鱼类种群的可持续利用和其生物量不会低到危害其生存,主要包括Fmsy、Fmax、F0.1、Fcrash、F20%、Bloss等参数。阈值参考点介于目标参考点和限制参考点之间,包括预防性捕捞死亡率Fpa、预防性生物量Bpa,主要对渔业资源的开发和管理进行预警,防止生物量小于BL。生物学参考点主要应用动态综合模型、产量模型和亲体量补充量关系模型来估算,估算过程中需要考虑到补充、生长、死亡等生命史过程中的不确定性。本文对生物学参考点的发展和应用进行了综述,并以金枪鱼渔业为例阐述它在渔业管理上的应用。近几十年来,我国近海渔业资源出现衰退,亟需利用生物学参考点的原理和方法对重要渔业资源种类进行...     

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。     

Infection routes of Aeromonas salmonicida in rainbow trout monitored in vivo by real‐time bioluminescence imaging

Recent development of imaging tools has facilitated studies of pathogen infections in vivo in real time. This trend can be exemplified by advances in bioluminescence imaging (BLI), an approach that helps to visualize dissemination of pathogens within the same animal over several time points. Here, we employ bacterial BLI for examining routes of entry and spread of Aeromonas salmonicida susbp. salmonicida in rainbow trout. A virulent Danish A. salmonicida strain was tagged with pAKgfplux1, a dual‐labelled plasmid vector containing the mutated gfpmut3a gene from Aequorea victoria and the luxCDABE genes from the bacterium Photorhabdus luminescens. The resulting A. salmonicida transformant exhibited growth properties and virulence identical to the wild‐type A. salmonicida, which made it suitable for an experimental infection, mimicking natural conditions. Fish were infected with pAKgfplux1 tagged A. salmonicida via immersion bath. Colonization and subsequent tissue dissemination was followed over a 24‐h period using the IVIS spectrum imaging workstation. Results suggest the pathogen's colonization sites are the dorsal and pectoral fin and the gills, followed by a progression through the internal organs and an ensuing exit via the anal opening. This study provides a tool for visualizing colonization of A. salmonicida and other bacterial pathogens in fish.     

凡纳滨对虾不同生长阶段肠道可培养细菌耐药性研究

在凡纳滨对虾(Litopenaeus vannamei)养殖过程中多使用微生态制剂来调节水质,为避免破坏池塘菌群结构,很少使用抗生素.为了解凡纳滨对虾肠道细菌耐药性与不同生长阶段的关系,本研究选取江苏地区4种主要养殖模式凡纳滨对虾成虾和虾苗作为研究对象,利用K-B纸片法和qRT-PCR技术,研究对虾样本肠道可培养细菌对...     

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.     

Effects of Estradiol and Testosterone on the Expression of Growth‐related Genes in Female and Male Nile Tilapia,Oreochromis niloticus

Nile tilapia exhibits strong sexual growth dimorphism. The potential role of sex steroid hormones in sexual growth dimorphism is not fully understood. We investigated the effects of estradiol (E₂) and testosterone (T) on growth rate, plasma sex hormones, and expression of growth hormone (GH)‐insulin‐like growth factor (IGF) axis genes and muscle regulatory factor (MRF) genes in female and male Nile tilapia. The results revealed that serum concentrations of E₂ and T were significantly higher after correlative injection (P < 0.05). Compared to male fish, female fish had lower growth rates. E₂ increased growth performance in females with no significant effects on males, whereas T significantly increased growth performance in males, with no significant effects on females. In females, E₂ significantly increased expression of ghr1, ghr2, igf1, and igf2, while T decreased igf2 and increased ghr1 and ghr2 expression. In males, T increased expression of igf1, igf2, ghr1, and ghr2, and E₂ decreased expression of igf1, ghr1, and ghr2. Additionally, E₂ and T enhanced the expression of MRF genes (myod1, myod2, myog, and myf5) in female and male fish, respectively. The results suggest that sex steroid hormones play a role in sexual growth dimorphism by regulating the expression of GH‐IGF axis and MRF genes.     

山东近海口虾蛄单位补充量渔获量评估

为完善口虾蛄的基础生物学资料,并为口虾蛄资源的管理提供科学指导和理论依据,本研究根据2016至2017年山东近海渔业资源底拖网调查获得的口虾蛄体长、体质量数据,估算了口虾蛄的生长、死亡参数,构建了基于体长结构的单位补充量渔获量(YPR)模型,研究口虾蛄的资源动态和管理策略。采集调查口虾蛄样品共5 028尾,体长—体质量关系的表达式为W=0.014 5L2.88,为负异速生长;使用ELEFAN方法估算出口虾蛄的渐进体长L∞为19.87 cm,生长速率K为0.62 a-1。口虾蛄的生长表现出明显的季节性变化规律,生长参数的季节振幅C为0.76,10月份生长最快,4月份生长最慢。通过体长转换的渔获曲线估算出口虾蛄的总死亡系数Z为3.24 a-1,根据不同方法估算自然死亡系数M的范围为0.75～1.27 a-1,捕捞死亡系数F的估算范围为1.96～2.49 a-1,开发率的均值为0.67。YPR模型结果显示,随着F增大,YPR值呈现先上升后下降的趋势,生物学参考点F0.1和Fmax的值分别为0.92a-1和1.88a-1。口虾蛄资源处于过度开发的状态,应降低捕捞压力,同时调整开捕体长,以维持口...     

刺参响应灿烂弧菌侵染差异microRNAs鉴定及靶基因分析

microRNA参与基因的转录后调控,在真核生物的生长发育、细胞分化和免疫防御等过程中发挥重要作用。刺参(Apostichopus japonicus)病害问题已成为产业发展的主要限制因素之一,而其病害发生的分子机制尚待进一步完善。本研究以刺参重大疾病“腐皮综合征”的重要致病原灿烂弧菌(Vibrios splendidus)为侵染菌株,通过人工侵染实验制备患病刺参样本,采用miRNA-seq技术对侵染组(PT16S)和对照组(PT10H)各3头刺参的体壁组织进行miRNA测序,通过相关生物信息学软件对miRNAs进行鉴定和分析,筛选差异表达miRNAs (DEmiRNAs)并预测其靶基因,构建关键调控途径的miRNA-mRNA调控网络。结果显示,PT10H组平均得到5 902 588条有效序列,194个已知miRNA和19个新的miRNA;PT16S组平均得到5 053 529条有效序列,182个已知miRNA和42个新的miRNA。对2组鉴定到的miRNA进行差异表达分析,共筛选到2个上调和11个下调的具有显著差异的DEmiRNAs (P≤0.05),上调的DEmiRNAs靶基因预测结合到3010个靶基因,注释到585个GO terms及24条信号通路(P≤0.05),下调的DEmiRNAs靶基因预测到19 072个靶基因,注释到514个GO terms以及22条信号通路(P≤0.05)。对筛选到的DEmiRNAs进行实时荧光定量PCR (qRT-PCR)验证,显示miRNA-seq与qRT-PCR的一致率达到70%。根据KEGG分析结果构建泛素介导的蛋白水解途径和Notch信号通路的miRNA-mRNA调控网络,结果显示,13个DEmiRNAs分别靶向结合134个与泛素介导的蛋白水解相关的mRNAs和109个与Notch信号通路相关的mRNAs,Aja-miR-184、Aja-miR-2478和Aja-miR-9277p等DEmiRNAs可能参与对Notch信号通路和对泛素介导的蛋白水解的调控。相关研究结果将为刺参疾病发生调控网络建立和机制解析提供依据。