异育银鲫寄生洪湖碘泡虫的鱼卵传播途径

洪湖碘泡虫(Myxobolus honghuensis)是引起异育银鲫(Carassius auratus gibelio Bloch)“喉孢子病”的重要病原, 每年导致养殖苗种和成鱼大量死亡。本研究通过隐性感染异育银鲫母本人工受精、实验室条件下受精卵孵化和幼鱼培育, 采用单管半巢式 PCR、荧光定量 PCR 和寡核苷酸荧光原位杂交等检测手段进行亲本、卵和幼鱼等环节的检测分析, 探究异育银鲫寄生洪湖碘泡虫是否存在经卵传播途径。结果表明, 所采用的 34 尾异育银鲫母本(A1~A22, B1~B12)的洪湖碘泡虫隐性感染率达 50%~75%, 其中, 卵和伪鳃检出率高于卵巢组织样品; 特异性寡核苷酸探针荧光原位杂交在隐性感染母本的卵巢、伪鳃、肾、脾组织检测到洪湖碘泡虫前孢子生成阶段营养体; 实验室条件下阳性母本所产的卵经孵化和培育出的幼鱼 15 dph 和 30 dph 样品可以检出阳性(A1、A18、B8 和 B9); 荧光原位杂交显示 15 dph 幼鱼在伪鳃、鳃和肾脏组织检测出阳性信号。本研究进一步揭示了异育银鲫寄生洪湖碘泡虫存在经鱼卵传播途径; 研究结果可为相关疾病制定防控措施奠定重要的理论基础。     

添加酶制剂对异源银鲫苗种培育的影响试验

异源银鲫,又名异育银鲫,它是应用细胞工程和有性杂交相结合培育出的三倍体鲫鱼,即由东北鲫鱼与珠三角的鲤鱼杂交而成,它具有自身不育、食性广、生长快、抗病能力强、广温性、耐低氧、肉质鲜美等特点,在珠三角地区是一种值得推广的养殖新品种.     

甲苯咪唑在银鲫体内的药代动力学研究

采用高效液相色谱法检测甲苯咪唑(MBZ)及其代谢物氨基甲苯咪唑(MBZ-NH2)和羟基甲苯咪唑(MBZ-OH)在银鲫(Carassius auratus gibelio)血浆及组织中的浓度,数据经3P97药代动力学程序分析.结果表明:(25±1)℃的水温条件下,银鲫单剂量口灌MBZ20mg·kg-1,血药经时过程符合二室开放式模型.主要药代动力学参数为:吸收速率常数恐Ka 0.235h-1;消除半衰T(1/2)β 52.26h;药时曲线下面积AUC 180.07μg·h·mL-1;表观容积分布Vd/F 2.465L·kg-1;清除率CL(s)0.111 mL·h-1·kg-1;达峰时间T(peak) 6.20 h;质量浓度Cmax 4.14 μg·mL-1.与哺乳类相比,MBZ在银鲫体内吸收较慢,消除半衰期明显延长,药代动力学参数也有较大差异.建议在做到合理用药的同时加强药物检测力度.     

银鲫多核苷酸重复微卫星的序列同源性分析

应用磁珠富集法开发银鲫的三、四核苷酸重复的微卫星。生物素探针分别为Probe-(TGA)8,Probe-(AGAC)8,Probe-(GATT)8,结果得到4740个菌落。结合同位素放射法进行二次筛选,获得652个阳性克隆(阳性克隆率13.76%),利用PRIMER3PLUS在线软件设计引物163对,采用in silico方法,对微卫星序列进行BLASTN和BLASTX在线搜索。运用BLASTN搜索,首先将设计引物后的微卫星序列(侧翼序列长度大于50bp)与GEN-BANK中的斑马鱼的EST数据库进行比对,共匹配上44个同源序列。然后利用BLASTX搜索非冗余蛋白数据库,有31个微卫星位点成功的进行了功能注释,5个微卫星位点匹配上假设蛋白。本文利用模式生物斑马鱼的基因组资源对银鲫的的基因组进行比较研究,为以后银鲫种质资源的开发打下了良好的基础。     

Induced diploid gynogenesis and polyploidy in crucian carp, Carassius auratus gibelio (Bloch), x common carp, Cyprinus carpio L., hybrids.

Abstract. Results of a 16 year (1977–1992) study of hybrids obtained from crossing two cyprinid fish, crucian carp, Carassius auratus gibelio (Bloch) (bisexual form), and common carp, Cyprinus carpio L., are presented. The ability to produce diploid eggs in F₁ females (resulting from chromosome endoreduplication in early oogenesis) was responsible for the high yield of spontaneous diploids in gynogenetic progenies of F₁ females and for triploidy in hybrids from back-crosses of F₁ females with males of the parent species. Different types of hybrids, including diploid and triploid gynogenetic, tetraploid (amphidiploid) and androgenetic progenies, were obtained. Diploid and some triploid hybrids (females only) were reproduced inter se by induced gynogenesis in several subsequent gynogenetic generations. Amphidiploid hybrids were obtained by crossing triploid F₈ females with diploid common carp males or by crossing diploid gynogenetic females with diploid inverted hybrid males producing diploid sperm. Results of cytogenetic investigations and fish culture properties in hybrids are presented. The scientific and practical significance of the data obtained are discussed.     

浅山区塘坝养殖方正银鲫试验研究

2002年4月10日～2002年10月15日在方正县选择浅山区塘坝进行方正银鲫养殖试验研究。结果表明：在低温浅山区塘坝进行方正银鲫为主的养殖是可行的，经：170d的养殖，方正银鲫由放养时的l00g到出池时250g，亩利润为1233．4元，同时，搭配大规格鲢鳙鱼，可取得高产高效的养殖效果。     

Evaluation of rendered animal protein ingredients for replacement of fish meal in practical diets for gibel carp,Carassius auratus gibelio (Bloch)

A 12‐week feeding trial was carried out in fibreglass tanks to examine partial replacement of fish meal (FM) with poultry by‐product meal (PBM), meat and bone meal (MBM) and blood meal (BM) in practical diets for gibel carp Carassius auratus gibelio (Bloch). Triplicate groups of fish (initial body weight 15.3 g fish^?1) were fed eight isonitrogenous (crude protein: 37.5%) and isolipidic diets (crude lipid: 7%). The control diet is the commercial diet of gibel carp, which used 18% of FM as the sole animal protein source. In the other seven diets (Diet 2–Diet 8), 17–83% of FM protein was substituted by a blend of PBM and BM or a blend of PBM, MBM and BM. The final body weight and thermal‐unit growth coefficient of fish fed the feeds in Diet 8 in which 83% of FM protein was replaced by the blend of 3% BM, 10% PBM and 5% MBM is significantly lower than Diet 1 (the control). The feed efficiency ratio in Diet 8 group is significantly lower than Diet 1, 2, 4 and 7 groups. The results of the present study indicated that a combination of PBM, BM and MBM can replace most of the FM protein and the FM level can be reduced to about 6% with satisfactory growth and feed utilization in practical diets for gibel carp.     

Thelohanellus testudineus n. sp. (Myxosporea: Bivalvulida) infecting the skin of allogynogenetic gibel carp Carassius auratus gibelio (Bloch) in China

A Thelohanellus species was encountered during a survey on Thelohanellus diversity of Carassius auratus gibelio (Bloch) in China. The infection is characterized by the presence of large cysts of 1.4–3.2 cm in diameter in the skin of host. Mature spores were ampullaceous in frontal view and testudinate in lateral view, measuring 19.7 ± 0.7 (18.6–20.8) μm long, 7.6 ± 0.4 (6.6–8.4) μm wide and 7.3 ± 0.5 (6.6–8.8) μm thick. The single polar capsule was elongated pyriform, with 11.1 ± 0.5 (10.0–11.9) μm long and 5.3 ± 0.3 (4.3–5.8) μm wide. Polar filaments coiled with 7–8 turns. Scanning electron microscopy revealed a smooth spore surface with flat side and convex side. The sutural line was straight or ‘S’ like, running near the middle of the valves. Histologically, the large cysts consisting of numerous small plasmodia developed in the dermis of the skin. The BLAST search indicated that the newly obtained ssrRNA gene sequences did not match any available sequences in GenBank and phylogenetic analysis placed it in the Thelohanellus clade. Based on morphology and molecular differences with reported Thelohanellus spp., this parasite was described as a new species of genus Thelohanellus.     

Partial and total replacement of fishmeal with poultry by-product meal in diets for gibel carp, Carassius auratus gibelio Bloch

Triplicate groups of gibel carp Carassius auratus gibelio Bloch (initial body weight: 4.89 g) were fed for 8 weeks at 24.8–30.8°C with nine isonitrogenous and isoenergetic diets. The control diet (F1) used white fishmeal (FM) as the sole protein source. In the other eight diets (F2–F9), 40.5–100% of FM protein was substituted by poultry by‐product meal (PBM) at 8.5% increments. The specific growth rate (SGR), feed efficiency ratio, protein efficiency ratio, protein retention efficiency and energy retention rate for fish fed PBM diets (F2–F9) were all higher, but not always significantly, than those for fish fed F1. All apparent digestibility coefficients for fish fed PBM diets were lower than those for fish fed F1. Fish fed F1 had a significantly higher hepatosomatic index value than fish fed PBM diets (P<0.05). No significant (P>0.05) effect of diet was found in whole‐body moisture and fat content. Whole‐body protein and energy content for fish fed PBM diets were slightly higher than that for fish fed F1. The optimal replacement level of FM by PBM was estimated by second‐order polynomial regression to be 66.5% in protein.     

Effect of partial replacement of dietary monocalcium phosphate with neutral phytase on growth performance and phosphorus digestibility in gibel carp,Carassius auratus gibelio (Bloch)

A feeding trial was conducted to study the effect of partial replacement of dietary monocalcium phosphate (MCP) with neutral phytase on growth performance and phosphorus digestibility in gibel carp, Carassius auratus gibelio (Bloch). Control diet was prepared with 2% MCP but without phytase (P0). Other three experimental diets were prepared by replacement of MCP by 25%, 50% and 75% respectively in comparison with control with supplementation of neutral phytase at 500 U kg^?1 diet in each and designated as P25, P50 and P75 respectively. Gibel carp (initial body weight of 30.22 ± 1.98 g) were reared in twelve 300‐L cylindrical fibreglass tanks provided with filtered flow‐through tap water at 26–28°C. After 8‐week experiment, gibel carp fed with P50 had no obvious differences from the control group on weight gain (WG), specific growth rate (SGR), feed conversion ratio (FCR), protein efficiency rate (PER) and survival rate. Phytase supplementation did not affect body compositions or muscle compositions. Crude protein and phosphorus (P) contents in the faeces of fish fed with the phytase‐supplemented diets were significantly lower than those of the control group. The apparent digestibility coefficients (ADCs) of crude protein and P in gibel carp were increased when fish fed with the diets in which MCP was replaced by neutral phytase. This study suggested that partial replacement of dietary MCP at 50% with neutral phytase was considered as a recommended dietary supplemental level and increased dietary P and protein availability.     

磁珠富集法筛选方正银鲫的三、四核苷酸重复微卫星DNA

通过生物素磁珠富集法开发方正银鲫(Carassius auratus gibelio (Bloch))三、四核苷酸重复的微卫星,并与传统的放射性同位素杂交法相结合,进行二次筛选微卫星分子标记.结果显示:对652个阳性克隆进行测序分析,获得微卫星序列800个,其中完美型542个(67.75%),非完美型140个(17.5...     

Dietary myo‐inositol requirement for juvenile gibel carp (Carassius auratus gibelio)

An 11‐week growth trial was conducted to determine dietary myo‐inositol (MI) requirement for juvenile gibel carp (Carassius auratus gibelio). Myo‐inositol was supplemented to the basal diet to formulate six purified diets containing 1, 56, 107, 146, 194 and 247 mg MI kg^?1 diet, respectively. Each diet was fed to triplicate groups of juvenile gibel carp (initial body weight 3.38 ± 0.27 g, mean ± SD) in a flow‐through system. The diets were randomly assigned to different fish tanks. Fish fed ≥ 107 mg MI kg^?1 diet had significantly higher weight gain (WG), feed efficiency (FE) and protein efficiency ratio than those fed 1 mg MI kg^?1 diet. Fish fed ≥ 56 mg MI kg^?1 diet had higher feeding rate and survival compared with fish fed 1 mg MI kg^?1 diet. Dietary supplemental inositol did not affect fish liver inositol concentration. Fish fed ≥ 56 mg MI kg^?1 diet had higher body dry matter, crude protein and gross energy and lower hepatosomatic index than fish fed 1 mg MI kg^?1 diet. Dietary inositol supplementation decreased fish body ash. Quadratic regression of weight gain indicated that the myo‐inositol requirement to maximum growth for juvenile gibel carp was 165.3 mg MI kg^?1 diet.     

分级对银鲫生长和个体大小差异的影响

在室内水槽进行12周的饲养实验以检验分级对银鲫生长和鱼群内个体大小差异的影响。用隔网将实验水槽分为两部分,每个水槽内放养10尾大鱼（体重8.0~9.0 g）和10尾小鱼（体重6.5~7.5 g）,分级时将个体大小相近的10尾鱼分布在隔网同一侧。实验中,3组鱼分别隔1周、2周或4周分级1次,1组鱼不进行分级作为对照。鱼摄食率和生长率随分级间隔时间缩短而降低。实验结束时,隔1周分级1次的鱼体重低于对照组,隔4周分级1次的鱼体重与对照组无显著差异。实验结束时被分级的鱼群中大鱼体重的变异系数高于小鱼,表明大鱼对分级操作更敏感。本实验结果表明,分级未能明显提高银鲫生长速度和降低鱼群内个体大小的差异程度。     

黄芩苷与甘草酸对恩诺沙星在异育银鲫体内代谢的影响

将黄芩苷(baicalin,BL)和甘草酸(glycyrrhizin,GZ)口灌异育银鲫(Carassius auratus gibelio),探讨其对恩诺沙星(enrofloxacin,ENR)在体内的代谢和肝微粒体细胞色素氧化酶CYP1A、CYP3A活性的影响。异育银鲫连续7 d分别口灌黄芩苷(100 mg/kg)和甘草酸(100 mg/kg),以口灌玉米油作为对照。末次给药24 h后每组随机取10尾腹腔注射恩诺沙星(10 mg/kg),采用单个动物连续采血,HPLC测定血浆恩诺沙星和其代谢产物环丙沙星(cipmfloxacin,CIP)浓度,分析药动学及其参数;同时每组选取6尾检测肝微粒体CYP1A和CYP3A活性。结果表明:(1)黄芩苷(BL)和甘草酸(GZ)对恩诺沙星的吸收有明显抑制作用,主要表现在恩诺沙星峰浓度(Cmax)降低,曲线下面积(AUC)减少;(2)口灌BL和GZ后,恩诺沙星及其代谢产物环丙沙星的消除半衰期(t1/2z)明显小于对照组(P<0.05),而总体清除率(CLz/F)则增大,说明黄芩苷和甘草酸促进了恩诺沙星和环丙沙星的消除;(3)口灌BL和GZ后,BL组和GZ组的Cmax-CIP/Cmax-ENR比值分别为1.48%、2.22%,对照为0.95%;BL组和GZ组的AUC0-t-CIP/AUC0-t-ENR比值分别为2.16%、1.76%,对照组为1.7%。综合分析代谢产物环丙沙星峰浓度、Cmax-CIP/Cmax-ENR和AUC0-t-CIP/AUC0-t-ENR比值可以得出,黄芩苷和甘草酸对恩诺沙星N-脱乙基具有诱导作用;(4)与对照组相比较,BL组和GZ组的7-乙氧基异吩唑酮-O-脱乙基酶(EROD,CYP1A标志酶)和红霉素-N-脱甲基酶(ERND,CYP3A标志酶)活性显著升高(P<0.05),说明黄芩苷和甘草酸对CYP1A和CYP3A都有诱导作用。结合以上结果,认为黄芩苷和甘草酸加速了恩诺沙星的消除和其代谢产物CIP的生成,很可能与诱导CYP1A和CYP3A活性有关。     

黑龙江银鲫新品系—松浦鲫的培育

以纯种方正鲫作母本、以方正鲫(♀)与鳞鲤(♂)交配获得的雌性后代经性反转的生理雄性个体作父本进行育种试验,在3个组配中,发现有一组F_1表现出新的形态特征。经过对F_1及其后代作定向培育和观测,证实F_1形态指标与方正鲫差异显著,且其后代遗传性状稳定,可作为鲫鱼属的一个新品系。     

银鲫crh基因的克隆、组织表达谱及其对摄食的影响

为研究crh (Corticotropin-releasing hormone)基因对鱼类摄食的调控作用,首次克隆了银鲫(Carassius auratus gibelio) crh基因c DNA全长序列。运用实时荧光定量PCR(RT-qPCR)技术检测crh基因m RNA在不同组织的表达及餐前、餐后和禁食对其表达的影响。结果显示,银鲫crh基因的c DNA序列全长为920 bp,其中,5’-UTR为53 bp,3’-UTR为378 bp,开放阅读框(Open reading frame,ORF)为489 bp。推导银鲫crh基因的ORF区编码蛋白由162个氨基酸组成,其中,含有11个氨基酸的保守区,24个氨基酸的信号肽,41个氨基酸的成熟肽。氨基酸序列的多重比较分析显示,银鲫crh基因与金鱼(Carassiusauratus)的同源性为99%,与鲤(Cyprinuscarpio)的同源性为96%。荧光定量PCR表达分析显示,银鲫crh基因在下丘脑中的表达量最高,其次是端脑和心脏。crh基因的表达量在餐前与餐后没有显著变化(P0.05),在禁食第5天、第7天出现极显著降低(P0.01),而在复投喂后第9天、第11天出现极显著升高(P0.01)。以上结果显示,crh基因在银鲫摄食调控方面可能扮演着重要角色。