奥利亚罗非鱼（♀）、鳜（♂）及其子代间遗传关系的研究

罗非鱼生长快、味道好,是人们喜爱的鱼类,现已是世界性养殖鱼类,但它不能忍受低温,因此影响其养殖业的进一步发展。鳜则是名贵性鱼类,但其终生摄食活的鱼虾,这也限制了其养殖的发展。奥利亚罗非鱼(Oreochromisaurea)属鲈形目、鲡鱼科,鳜(Sinipercachuatsi)属鲈形目、脂科,它们间的杂交为科间杂交。鱼类科间杂交成功率很低,种间杂交有时可获得具有双亲性状的杂种鱼,有时也可发生雌核发育[1]。本研究中奥利亚罗非鱼雌鱼性染色体为ZW,其卵子如能在鳜精子刺激下行雌核发育,则可在短时间获得奥利亚罗非鱼纯系。在本文中,利用RAPD标记,对…     

Perkinsus sp. infestation in carpet-shell clams, Ruditapes decussatus (L), along the Portuguese coast. Results from a 2-year survey

This study characterises Perkinsus atlanticus disease prevalence and intensity within the economically important beds of the clam Ruditapes decussatus along the Iberian Atlantic coast. Samples were collected from five different sites along the Portuguese coast (Ria Formosa, Ria do Alvor, V.N. Milfontes, Aveiro, Lagoa de Obidos) and in Galicia (Ria de Arosa) between winter 2000/2001 and 2002/2003. The infection level was evaluated by the Ray Fluid Thioglycollate medium (RFTM) method using the body burden assay. In addition, the measure of the condition index (CI) (percentage between the edible part and the total weight of the clams) was conducted to investigate a possible correlation between this parameter and the intensity of the infection. However, no clear relationship between infection intensity and condition index (CI) could be determined although a significant decline in CI was found for heavily infected clams. Results of a 2-year survey demonstrate the presence of Perkinsus sp. in all sampling sites albeit with different intensities. Sites where human interference was minimal showed the lowest levels of infection. There were no significant differences in Perkinsus sp. infection intensity between samples collected in winter and summer, in contrast with several studies describing higher intensities at the end of the summer. Nevertheless, major differences were observed from year to year and site to site, indicating that factors other than those responsible for seasonal climatic variations might affect the prevalence and the intensity of Perkinsus sp. infection.     

河蟹青虾混养池塘循环水净化效能的初步分析

在河蟹青虾混养池塘构建封闭式循环水养殖系统,并对其净化效果和养殖效益进行了初步研究。结果表明：该系统对水体TN、TP、NH4＋-N和CODMn的平均净化效率分别为27.33%、56.14%、43.91%和39.59%。经湿地净化后的水质能够达到GB3838-2002地表水Ⅲ类排放标准,平均纯收益达53769元/hm2。     

Comparison of susceptibility of various fish species to experimental infection with channel catfish virus

Channel catfish virus (CCV) disease is an acute haemorrhagic disease in juvenile channel catfish ( Ictalurus punctatus ). To date channel catfish is the only species affected by natural outbreaks of the CCV but juvenile large mouth bass ( Micropterus salmoides ) and silurus ( Silurus meriaionalis ) have suffered high mortalities in recent years in China. Histopathological phenomenon of sick fish is similar to CCV disease, and the identified virus was CCV. In this report, the pathogenicity of infectious CCV was examined by infection trials on the first known host species, the channel catfish and other teleosts. Our results indicated that there were higher detection rates of CCV from large mouth bass and silurus fish. Channel catfish virus did not induce mortality in other cypriniformes, but histopathological studies revealed that carp might be infected by both bathing and intraperitoneal infection. No deaths, clinical or histopathological signs, were found in the six other species exposed by immersion or injection. Experimental infection studies confirm that CCV infect not only channel catfish but also other species (large mouth bass, silutus and carp). The outbreaks of CCV disease only occurred when the cultured temperature was above 25 °C.     

Development and application of fluorescence in situ hybridization (FISH) method for simple and rapid identification of the toxic dinoflagellates Alexandrium tamarense and Alexandrium catenella in cultured and natural seawater

ABSTRACT:   The toxic dinoflagellates Alexandrium tamarense (Lebour) Balech and A. catenella (Whedon and Kofoid) Balech produce potent neurotoxins, such as saxitoxin and gonyautoxin and have been mainly responsible for paralytic shellfish poisoning (PSP) in Japan. To prevent a negative effect on the fishery industry, it is necessary to identify these toxic species precisely and rapidly before and during the bloom. In this paper, a rapid and simple protocol of a fluorescence in situ hybridization (FISH) method using ribosomal RNA (rRNA)-targeted probes has been established for identifying the cultured strains and natural cells of A. tamarense and A. catenella . Using the FISH method established in this study, it was possible to identify these toxic species species-specifically and rapidly, within 30 min. The procedure of detection constituted three steps: (i) fixation/dehydration; (ii) hybridization; and (iii) washing; this made the identification simple. Moreover, this method did not require either special techniques or equipment, and the cost for detection was low. The specificity, rapidity, and simplicity of the developed method suggest that it might be useful for routine monitoring of these toxic microalgae.     

长江草鱼群体MHC Class ⅡB的多态性和进化分析

克隆及测序草鱼(Ctenopharyngodon idella)长江3个群体的主要组织相容性复合体(MHC)Class II B基因编码β1和β2区的第2和第3个外显子及两个外显子之间的内含子,分析了草鱼MHC的进化模式和种群遗传结构。结果显示:实验共定义了34个等位基因,每条序列包括长为130~136 bp的第2个外显子,长为218 bp的第3个外显子以及长81~371 bp的内含子。序列分析揭示,第2个外显子有106个核苷酸变异位点(78%)和40个氨基酸变异位点(88%),而第3个外显子有100个核苷酸变异位点(45%)和41个氨基酸变异位点(56%),β1变异要大于β2区。用β1和β2区序列分别构建的邻接(NJ)系统树均显示5个具有高支持率的谱系,结合序列变异特点和内含子长度,推测草鱼至少存在5个MHC Class II座位。分别计算β1的肽结合位点(PBR)、非肽结合位点及β2的非同义替换率(dN)和同义替换率(dS),PBR的dN/dS为2.03(P<0.05),非肽结合位点和β2则小于1,表明草鱼MHC受到歧化选择作用。根据等位基因在群体中的分布频率作分子方差分析(AMOVA),得出FST为0.37%,提示长江草鱼MHC没有遗传分化。     

草鱼HSP70基因cDNA部分序列克隆及其组织表达差异

根据鲤热休克蛋白70(Heat shock protein,HSP70)序列(AY120894)设计并合成一对引物,以草鱼(Cteno-pharyngodon idella)肝胰脏组织总RNA为模板,RT-PCR扩增获得草鱼HSP70基因cDNA部分序列,并进行了组织表达差异性研究。结果显示:所获为序列为480 bp,获得GeneBank登陆号为FJ483832。序列测序结果显示,HSP70扩增序列与鲤、斑马鱼、鲋的同源性为:93%、91%、93%。另外,所获序列HSP70在草鱼脂肪、肌肉、肠、脑、粘液、性腺、鳔、肝胰脏、心脏、脾脏、鳃、鳍12个组织的表达存在差异,HSP70在草鱼这12个组织中均检测到表达,其中在鳍中表达最高,极显著高于其他组织(P<0.01);在鳔中表达次之,且与脑、心脏、性腺中表达差异不显著;在粘液中表达最低。     

栉孔扇贝BES-SSR的开发及遗传多样性分析

通过分析栉孔扇贝BAC末端序列,发现大量微卫星DNA;随机选择14个多态性BES-SSR标记,在我国栉孔扇贝大连群体(DL)和青岛群体(QD)中验证标记的可用性,同时对这两个群体的遗传结构及其分化进行研究。结果表明,从17447条BESs中得到微卫星3374个,以四核苷酸重复为主(26.6%),五核苷酸重复次之(17.7%),六核苷酸重复最少(12.0%)。BES-SSR引物的扩增效率为77.3%(99/128),在作图亲本中的多态比例为33.6%(43/128),14个基因座在两群体中的平均等位基因数Na分别为18.9286和26.2143,平均有效等位基因数Ne为11.7505和17.0891,平均观察杂合度Ho为0.5100和0.4204,平均期望杂合度He为0.9156和0.9450,多态信息含量PIC分别为0.8940和0.9302,群体遗传多样性水平较高。两群体间的无偏遗传相似性系数为0.4879,遗传距离为0.7177,平均基因分化指数FST为0.0243,基因流Nm为10.0179,显示群体间遗传分化程度较弱,遗传变异主要来自于群体内个体之间,经Hardy-Weinberg平衡检验,两群体普遍存在杂合子缺失现象。研究表明,所开发的BES-SSR是高度多态位点,用于群体遗传多样性分析效果很好,显示BES是微卫星标记开发和应用的重要资源。     

Dominant sources of dietary carbon and nitrogen for shrimp reared in extensive rice-shrimp ponds

Stable isotope analysis was used to determine the sources of dietary nitrogen (N) and carbon (C) for shrimp during the rearing phase in extensive rice‐shrimp ponds in My Xuyen and Gia Rai districts, Vietnam. Farm‐made feed was added as a food source in shrimp ponds in My Xuyen district, and based on stable isotope analyses, was generally a poor dietary source. The commercial formulated feed used in Gia Rai also appeared to contribute little directly to the nutritional needs of the shrimp. In contrast, the natural biota in all ponds appeared to contribute substantially. In particular, biota from beam trawls and benthic organic matter were the most likely sources of nutrition in My Xuyen ponds, while benthic organic matter was the main source in Gia Rai ponds. δ¹⁵N ratios in the natural biota in My Xuyen farms decreased over the growing season, suggesting increased N fixation in the case of the benthic organic matter reaching values as low as 1‰. This suggests N‐limitation in the ponds and that natural biota become increasingly dependent on N fixed by algae and/or other microorganisms. There is the potential to promote the growth of the plankton and hence, the other natural biota, by the judicious addition of fertilizer.     

栉孔扇贝、虾夷扇贝及其杂交子代的MSAP分析

运用甲基化敏感扩增多态性(methylation-sensitive amplification polymorphism,MSAP)技术对栉孔扇贝(♀)、虾夷扇贝(♂)及其杂交子一代、子二代基因组DNA胞嘧啶甲基化水平进行了研究,并分析了DNA甲基化与各性状的相关性及其与杂种优势的关系。结果表明,(1)DNA甲基化率与壳宽、总重等表型值呈正相关的关系,而与壳长、壳高、软体重和闭壳肌重4个性状表型值呈负相关的关系,其中闭壳肌重与甲基化率的相关性达到极显著水平(P<0.01);(2)虾夷扇贝、栉孔扇贝、F1代、F2代的总甲基化率分别为32.79%、24.13%、19.98%、20.18%,杂交种F1代的甲基化水平低于双亲,是两种扇贝杂交的结果;F1代的甲基化模式经过了重新调整,其变化相对其亲本主要有4种类型:甲基化水平相同、去甲基化、超甲基化、次甲基化,且去甲基化位点多于超甲基化位点。结果证实杂种优势的产生与杂交种F1代基因组DNA甲基化模式的改变和重新调整有关,丰富了杂种优势机理研究内容。