利用肠道菌群的分布和DNA序列对鲤科鱼类系统演化关系的研究

用检测到的肠道微生物菌群的含量对鲤科鱼类的草鱼（Ctenopharyngodon idellus)和青鱼（Mylopharyngodon piceus)、鲢（Hypophthalmichthys molitrix)和鳙（Aristichthys nobilis)、鲤（Cyprinus carpiohaematopterus)和鲫（Carassius auratus)，团头鲂（Megalobrama amblycephala)和三角鲂（Megalopbrama terminalis）8个种进行系统演化分析。所获得的系统演化树中，每个亚科内的物种邻接为1个接点。在亚科间，雅罗鱼亚科与鳇亚科相邻领接1个共同接点。该接点与鲢亚科和鲤亚科连结为1个共同祖选接点。用线粒体基因中的cytb和ND4基因全序列分别构建5个亚科13和11个种的NJ树，这2个树在5个亚科之间的邻接拓扑结构上亦不相同。3种树可推出一个共同的结论是在所分析的亚科中鲤亚科为最原始的类群，而鲫是最原始的类型。     

Genetic parameters for growth‐related traits and survival with age in the Kuruma shrimp Marsupenaeus japonicus

Genetic parameters were estimated for growth‐related traits and survival of the Kuruma shrimp Marsupenaeus japonicus based on 66 families, including 30 paternal half‐sib families, which were obtained using artificial insemination of two females by each male. The variance components for growth‐related trait and survival were estimated using a single‐trait animal model and a sir‐dam model, respectively, and genetic parameters were estimated using the restricted maximum likelihood method. The coefficients of variation for growth‐related traits such as body length (BL), abdominal length (AL), and body weight (BW) were 9.36%–22.44%, 10.61%–21.92%, and 26.47%–58.33%, respectively, at different ages (45, 75, 105, 135, and 165 days). The corresponding heritability estimates for each growth trait were 0.1545–0.1951, 0.1672–0.1905, and 0.1596–0.1934, respectively, all of which were found to be at moderate levels and increased with age. The heritability of survival on day 165 was low (0.003). The genetic correlations among growth traits were positive and high (0.7316–0.9896) at the different ages, suggesting that selection to improve any single growth trait will cause positive responses in other growth traits examined in M. japonicus. The genetic correlations between growth traits and survival were also positive but low (0.005–0.087), which indicated that only selecting for growth traits may not cause a positive correlated response in the survival of the core population. According to the above results, we suggested that growth and survival traits should be taken as improving targets of breeding in M. japonicus. These results provided reference data for selective breeding and multitrait selection of M. japonicus.     

Stability comparison of three candidate internal reference genes in Chinese giant salamander (Andrias davidianus)

The Chinese giant salamander (Andrias davidianus) as food and medicinal product has been an important aquaculture object in China. Study of gene function in the Chinese giant salamander requires accurate normalization though the use of appropriate reference genes. In this study, the expression levels of three candidate reference genes including β‐actin, GAPDH and cytb of different tissues, different developmental stages and different challenges in Chinese giant salamander were evaluated by qPCR. The stabilities of these three reference genes were analysed by geNorm, NormFinder and BestKeeper software. The results showed that the expression of GAPDH was more stable than that of β‐actin and cytb in four tissues and at two developmental stages of Chinese giant salamander. Compared with GAPDH and cytb, β‐actin was the most stable in spleen of Chinese giant salamander treated with LPS or GSIV. Therefore, the result showed that GAPDH was the suitable reference gene in different tissues and at different developmental stages of Chinese giant salamander. The β‐actin could be used as a reference gene in spleen of Chinese giant salamander challenged with LPS and GSIV. This study provides convincing information for the GAPDH and β‐actin as suitable reference gene in Chinese giant salamander of different tissues, different developmental stages and different challenges respectively.     

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。     

水生生物近缘种和产地的分子生物学判别

随着分子生物学技术的发展,渔业生物近缘种及产地的判别由形态学深入到了蛋白质和DNA水平。本文综述了目前已经应用于水生生物的相关分子生物学技术(包括色谱、免疫学判别以及蛋白的电泳、DNA的序列分析、PCR-RFLP、RAPD、AFLP、SNP、SSRI、SSR、Real-Time PCRs、pecies-specific primer PCR和PCRlab-on-a-chip等)以及它们各自的优缺点;在把握了这些研究进展的基础上,展望了相关技术的发展趋势。