鳜→鲤的细胞核移植

本文报道用细胞核移植的方法,获得了亲缘关系较远的不同目鱼类之间的核一质杂种鱼胚胎.即以鳜鱼的囊胚细胞核作供体,以鲤鱼的成熟未受精去核卵细胞质作为受体进行核移植试验,使囊胚细胞核发生分裂,从而启动卵细胞质也发生分裂,导致重新进行个体发育.在获得的112个正常发育的核移植卵中,发生卵裂的107个,占95.5%发育至囊胚期的61个,占54.5%,发育至原肠期的6个,占5.4%,发育至神经胚期和胚孔封闭期的各1个,分别占0.9%.鳜→鲤核质杂种胚胎类似鲤鱼同期胚胎,其早期发育速度与鲤鱼相似而比鳜鱼快.各杂种胚胎发育期的胚盘形状及大小均类似鲤鱼,大于鳜鱼同期胚盘,且胚盘细胞较多,但比鲤鱼胚盘细胞小,这是鳜→鲤核质杂种胚胎发育的一个显著特点.作者认为核-质杂种胚胎的性状发育,受核受体物种的母性影响控制.     

Hematologic reference intervals for koi (Cyprinus carpio), including blood cell morphology, cytochemistry, and ultrastructure

BACKGROUND: Hematologic data are used routinely in the health care of humans and domestic mammals. Similar data for fish are largely fragmentary or have not been collected. OBJECTIVES: The primary purpose of this study was to determine hematologic reference intervals for koi, an ornamental strain of the common carp (Cyprinus carpio). Secondarily, the morphology, cytochemical reactions, and ultrastructure of koi blood cells were characterized. METHODS: A CBC was performed manually on heparin-anticoagulated blood specimens using Natt and Herrick's diluent and a Neubauer-ruled hemacytometer. Leukocyte differential counts were done on Wright-Leishman- and Diff-Quik-stained blood smears. Cytochemical reactions of koi leukocytes were determined using commercial kits. Transmission electron microscopy was performed to characterize the ultrastructural features of koi blood cells. RESULTS: Hematologic reference intervals were established for healthy koi for PCV (30-34%), hemoglobin concentration (6.3-7.6 g/dL), RBC count (1.7-1.9 X 10(6)/ microL), WBC count (19.8-28.1 X 10(3)/ microL), RBC indices, and differential leukocyte counts. Lymphocytes were the predominant leukocyte (accounting for up to 80% of all leukocytes), whereas eosinophils were rare. Basophils were positive with PAS staining. Naphthol AS-D chloroacetate esterase activity was observed only in eosinophils. alpha-Naphthyl butyrate esterase and beta-glucuronidase activities were positive in monocytes. Some lymphocytes were reactive for alpha-naphthyl butyrate esterase and acid phosphatase activity. Ultrastructurally, leukocytes, erythrocytes, and thrombocytes were identified on the basis of cytoplasmic organelles and granule appearance. CONCLUSION: Hematologic reference intervals and knowledge of the cytochemical reactions and ultrastructural characteristics of koi leukocytes will help standardize hematologic studies in this species.     

生长相关的微卫星标记在镜鲤繁殖群体中的验证

利用大样本镜鲤繁殖群体分析和验证8个生长相关微卫星标记与体质量和体长的相关性：结果显示：8个微卫星标记在镜鲤繁殖群体中均获得了稳定、清晰的DNA条带,片段大小在100-322bp之间,共检测到33个等位基因、75种基因型。对每个微卫星座位的基因型与体质量、体长性状进行相关性分析发现,HLJ343与体质量、体长显著相关;HLJ319、HLJ302及HLJE339与体长显著相关。本研究用大样本随机群体证实了此4个QTL位点与体质量或体长性状具有相关性,下一步可根据优势基因型指导家系配组,开展基于QTL结果的分子聚合育种研究。     

建鲤催乳素基因全长cDNA的克隆及序列分析和组织表达

为了解催乳素(prolactin,PRL)的基因序列以及在建鲤渗透调节组织中的表达情况,采用同源克隆和末端快速扩增(rapid amplication of cDNA ends,RACE)的方法分离克隆了建鲤(Cyprinus carpiovar.Jian) PRL基因全长cDNA,得到1 028 bp的全长cDNA,包括633 bp的开放阅读框(ORF),51 bp 的5′末端非编码区(UTR)以及344 bp的3′末端非编码区(UTR)。对该基因序列和推测的氨基酸序列进行同源性比对和系统分析显示：建鲤与其它硬骨鱼类该基因的氨基酸序列相似度为55.02%~94.76%,与草鱼(Ctenopharyngodon idella)相似度最高(94.76%),与齐口裂腹鱼(Schizothorax prenanti)、斑马鱼(Danio rerio)相似度稍低,与红鳍东方鲀(Takifugu rubripes)的相似度较低(55.02%),表明不同物种间PRL的氨基酸序列具有较高的保守性。用实时定量PCR (RT PCR)检测该基因在建鲤脑垂体、脑、肠、鳃、性腺、肝、脾、肾中PRL的相对表达量,其中脑垂体最高,其次是肝、鳃、脾、脑、肠、肾,在性腺中也有较低的表达量,表明脑垂体是建鲤PRL基因的主要表达场所,在性腺、肝、脾的表达说明其在鱼体内除了渗透调控可能还存在多种生理功能。     

“全红”和“粉玉”瓯江彩鲤体表组织中差异色素的鉴定

为了解"全红"和"粉玉"瓯江彩鲤(Cyprinus carpio var.color)体表组织中的差异色素,分别采用分光光度法和高效液相色谱方法对两种体色瓯江彩鲤的皮肤、鳍条和眼睛中的差异色素进行鉴定。分光光度法测定结果表明:"全红"皮肤中类胡萝卜素含量显著高于"粉玉"皮肤(P0.05),而蝶啶色素不存在显著性差异(P0.05)。随后通过高效液相色谱法对这两种体色的皮肤、眼睛和鳍条中类胡萝卜素的种类和含量进行鉴定,发现"全红"的皮肤、眼睛和鳍条中含有大量的以酯化形式存在的虾青素和黄体素,以及部分未知的类胡萝卜素,而"粉玉"的这些组织中检测不到虾青素和黄体素等类胡萝卜素。"全红"鳍条中虾青素含量最高为(28.00±3.61)mg/kg,而皮肤中的黄体素含量最高为(62.34±4.93)mg/kg。研究表明:虾青素和黄体素等类胡萝卜素是瓯江彩鲤红色体色的关键色素,蝶啶色素对红色没有作用,瓯江彩鲤白色体色存在类胡萝卜素的缺失现象。     

网箱养鳝模式对Cu2+的吸附和鱼生产性能的影响

为探讨网箱养鳝模式下空心莲子草和麦饭石对水体中Cu_²⁺的吸附效应和共生体系中黄鳝、鲤鱼存活率,增重率及肌肉Cu_²⁺含量的影响,试验选取健康黄鳝120尾,随机分成4组,每组设置3个重复,分别放养于网箱内;鲤鱼240尾,随机分成4组,每组设置3个重复,分别放养于网箱外水泥池中。饲养56 d。检测空心莲子草、麦饭石、水体Cu_²⁺含量和黄鳝、鲤鱼的存活率,增重率及肌肉Cu_²⁺含量。结果表明,空心莲子草和麦饭石对水体中的Cu_²⁺均有较强的吸附作用,水体Cu_²⁺浓度明显降低;随水体Cu_²⁺浓度升高,黄鳝和鲤鱼的存活率、增重率明显降低,肌肉中Cu_²⁺含量显著升高。     

Comparative functional responses of native and high‐impacting invasive fishes: impact predictions for native prey populations

Comparative functional responses (FRs) can predict impacts of invasive species, including piscivorous fishes, via quantifying their depletion of native food resources as a function of prey density. The utility of FRs for predicting impacts on prey populations by invasive fishes of different trophic guilds was tested here by comparing the FRs of the invaders Cyprinus carpio and Carassius auratus, with three native, trophically analogous fishes, Barbus barbus, Squalius cephalus and Tinca tinca. Chironomid larvae and Gammarus pulex were used as prey items. Predictions, developed from studies on the foraging of C. carpio and C. auratus in the literature, were that the invaders would have significantly higher consumption rates for chironomids than the native fishes, but not for G. pulex. Mean consumption rates for chironomids were significantly lower for both invaders than B. barbus and S. cephalus, but were similar to T. tinca. Barbus barbus had a significantly lower consumption rate of G. pulex than both invaders, but there were no significant differences between S. cephalus, T. tinca and the invaders. All FRs were type II, with FR curves for the invaders preying upon chironomids never being significantly higher than the native fishes, contrary to predictions. For G. pulex, some significant differences were apparent between the invaders and native fishes, but again were contrary to predictions. These results indicated that when predation impacts of invasive fishes could also be a function of their population density and body sizes, these parameters should be incorporated into FR models to improve impact predictions.     

饥饿和重摄食对鲤鱼脂蛋白脂肪酶活性的影响

采用对硝基苯酚法测定正常摄食、饥饿(14 d)和重摄食(3 d)状态下鲤鱼不同组织和器官中脂肪酶及脂蛋白脂肪酶(lipoproteinlipase,LPL)的活性。结果显示:与正常摄食相比,饥饿鲤鱼肌肉、脂肪和心脏中脂肪酶比活力显著下降(P0.05),但脂肪和心脏中LPL比活力显著升高(P0.05);除脂肪中LPL外,重摄食3d的鲤鱼各组织和器官中脂肪酶和LPL比活力比正常摄食和饥饿鲤鱼均显著升高(P0.05),有明显的补偿作用。     

鲤鱼肠道中4种细菌的分离与生化鉴定

为研究健康鲤鱼肠道菌群组成情况,为其饲料添加剂和水质改良剂的开发利用提供基础资料,试验对鲤鱼肠道中的细菌进行分离培养,通过形态特征观察、生化试验进行鉴定.结果:在鲤鱼肠道中分离出4种细菌,分别为:金黄色葡萄球菌(Staphyloccocus aureus)、沙门氏菌(salmonella)、粪肠球菌(Enterococcus faecalis)、枯草芽孢杆菌(Bacillus subtilis).     

从江县鲤爆发性死亡病例实验室诊断

2018年10月贵州省从江县某水产养殖专业合作社鲤爆发死亡,本研究对死亡病例进行了流行病学调查、病理学观察、寄生虫学检查、细菌分离鉴定和病毒PCR检测等诊断。结果显示:病鲤病变以鳃丝溃烂、体表出血、肛门红肿和眼球凹陷为主要特征,组织的主要病变是肾小球萎缩出血、肾小囊空腔、鳃丝肿胀粘连及上皮细胞脱落等;细菌分离培养可得到圆形凸起、表面光滑的灰白色菌落,经染色镜检、生理生化鉴定和16S rRNA基因测序分析,鉴定该分离菌为嗜水气单胞菌;PCR检测显示鲤浮肿病毒(Carp edema virus,CEV)为阳性,未检出鲤春病毒血症病毒(Spring viremia of carp virus,SVCV)和鲤疱疹病毒(Koi herpesvirus,KHV);显微镜观察亦未见到寄生虫虫体;分别取分离菌和鲤浮肿病毒阳性病例组织悬液进行人工感染试验,均可引起健康鲤发生死亡。上述结果表明,从江县鲤的死亡是嗜水气单胞菌和鲤浮肿病毒混合感染所致。