基于循环水养殖系统的长鳍吻鮈亲鱼培育、催产和孵化技术初探

2012—2015年,采用室内循环水养殖系统开展了长鳍吻鮈(Rhinogobio ventralis)亲鱼培育、人工催产和孵化技术研究。结果显示:3年间长鳍吻鮈均能在循环水养殖系统中培育成熟,2014年5月—2015年4月间培育存活率和成熟率最高,分别为85.3%~100%和77.3%~100%;对培育成熟的亲鱼进行了4种不同外源激素(PG、HCG、LRH-A2、LRH-A2+DOM)催产实验,结果为LRH-A26μg/kg+DOM 5 mg/kg合剂催产率最高,达88.3%;14、16、18、20和22℃5个温度组的受精卵孵化实验结果显示,孵化时间(H)与温度(T)呈显著负相关关系(H=0.573T2-30.393T+418.178,R2=0.982,P0.01),18℃和20℃组的孵化率分别为54.27%和56.07%,显著高于其他各组,表明长鳍吻受精卵适宜孵化温度范围为18~20℃。     

Comparison of in vitro vitellogenin syntheses by three phenols in primary cultures of Chinese minnow, Rhynchocypris oxycephalus hepatocytes

The in vitro effects of bisphenol-A (BPA), nonylphenol (NP) and 4-tert-octylphenol (OP) on vitellogenin (VTG) synthesis were examined using primary cultures of Chinese minnow, Rhynchocypris oxycephalus hepatocytes. In vitro VTG syntheses were induced by exposure to 10^?5 M of BPA and 10^?4 M of both NP and OP, suggesting that BPA had the highest estrogenic potential among the three phenols.     

长江宜宾江段圆口铜鱼遗传多样性的微卫星分析

选用实验室克隆的23个圆口铜鱼(Coreius guichenoti Sauvageet Dabry)微卫星标记分析了长江宜宾江段的圆口铜鱼群体遗传多样性,统计分析了有效等位基因数、观测杂合度Ho、期望杂合度He、多态信息含量(PIC)等遗传学指标。结果表明:23个位点有14个微卫星位点呈单态,9个位点出现多态,在这9个位点中共检测到48个等位基因,其平均有效等位基因数为5.3,多态信息含量在0.440～0.839之间变动,平均为0.670,除YT17和YT22位点属于中度多态外,其余7个位点均属于高度多态。平均观测杂合度为0.753,平均期望杂合度为0.728,表明该群体的遗传多样性较为丰富。     

长江天鹅洲故道银鮈年龄、生长和死亡率的研究

根据2011年3月至2012年1月在长江天鹅洲故道采集的银鮈(Squalidus argentatus)样本873尾,用鳞片为年龄鉴定和生长退算材料,研究银鮈的年龄、生长特性及死亡率.结果表明,雌雄群体均由1～6龄组成,雌鱼87.35％为1～3龄,雄鱼87.01％为1～3龄.雌雄鱼体长(L)和体重(W)关系表现为异速生长,雌鱼w=0.009L3.2285,雄鱼w=0.009L3.2254.以退算体长拟合von Bertalanffy方程,得到生长参数为:雌鱼L∞=17.1797cm,k=0.1820,t0=-1.2673和雄鱼L∞=11.5069cm,k=0.4074,to=-0.5764.雌雄鱼的自然死亡率分别为M=0.6048/a和M=1.1459/a,远高于一般鱼群自然死亡率(M=0.2～0.3/a);开发率分别为E=0.5711和E =0.4183.长江天鹅洲故道银鮈雌鱼处于过度开发的状态,应加强管理.     

紫薄鳅的人工繁殖初步试验

<正>紫薄鳅Leptobotia taeniaps(Sauvage)属鲤形目鳅科沙鳅亚科薄鳅属。分布于长江中、上游及其附属水体,属底栖性鱼类,喜生活在流水环境中,食物为底栖无脊椎动物。     

Hormone Profiles of Captive Striped Bass Morone saxatilis During Spermiation, and Long-Term Enhancement of Milt Production

Abstract— Plasma profiles of reproductive and thyroid hormones were studied in captive striped bass Morone saxatilis during an 11-wk period encompassing the spawning season, and the effect of a sustained-release gonadotropin-releasing hormone agonist (GnRHa)-delivery system (GnRHa-implant) on milt production was evaluated. The highest percentage of spermiating fish was observed between mid-April and mid-May, and mean total expressible milt ranged from 3.5 to 6.0 mL/kg. Plasma gonadotropin II (GtH II) increased significantly, though inconsistently, during the spermiation period, whereas testosterone and 11-ketotestosterone levels declined continually. Plasma 17,20β-dihydroxy-4-pregnen-3-one and 17,20β,21-dihydroxy-4-pregnen-3-one remained low and unchanged during the peak of the spermiation period, while thyroid hormones were high and fluctuated without exhibiting a trend consistent with spermiation. The observed endocrine profiles suggest that captivity can diminish plasma GtH II and triiodothyronine levels in striped bass. Transfer of spermiating males from large holding tanks to small spawning tanks reduced total expressible milt after 14 d, but treatment with a GnRHa-implant restored milt volume, presumably due to the prolonged elevation of plasma GnRHa and GtH II induced by the GnRHa-implant. Also, treatment with the GnRHa-implant induced a two- to four-fold elevation of expressible milt for at least 20 d compared to control fish, while resulting in only a 5 to 15% decrease in sperm density. It appears that captivity and hatchery operations can diminish milt production in striped bass, and that GnRHa-delivery systems, via sustained elevation of plasma GtH II, can induce long-term enhancement in milt volume without affecting sperm density greatly.     

Gynogenesis and sex determination in large-scale loach Paramisgurnus dabryanus (Sauvage)

Gynogenesis was induced using heterologous sperms in large-scale loach, Paramisgurnus dabryanus (Sauvage), in which a ZW/ZZ sex determination was previously proposed. Three microsatellite loci were used to monitor exclusive maternal inheritance of gynogenetic progenies. The results showed that high percentages of meiogynogens were produced at 4 min post-fertilization and mitogynogens were produced at 18 min post-fertilization by heat shocks, while meiotic gynogenesis was induced by cold shocks within a wide period and high heterozygosity was even observed in gynogens produced at 24 min post-fertilization. The sex ratios of the F₁ progenies in three gynogenetic families were significantly deviated from 1:1 expectation with a female bias in two families and a male bias in one family (P < 0.05), and the other four gynogenetic families showed approximate 1:1 sex ratios. Moreover, the self-mating between gynogenetic F₁ progenies and mating between gynogenetic F₁ progenies and normal individuals produced all-female progenies or identical proportions of females and males. The data of sex ratios generally confirmed that the sex determination in large-scale loach was determined by the putative ZW/ZZ system, and the possible reasons causing the biased sex ratios are discussed.     

南四湖大鳞副泥鳅生物学特性及人工繁育技术试验

为建立大鳞副泥鳅(Paramisgurnus dabryanus)的科学投喂策略,在实验室内开展了大鳞副泥鳅的养殖模拟实验,通过72 h的观察,探讨了不同体质量和不同性别大鳞副泥鳅的昼夜摄食节律及日摄食量。结果表明,大鳞副泥鳅属于典型的夜间摄食类型,一天中有2个极明显的摄食高峰,集中在04:00—05:00和20:00—21:00。大鳞副泥鳅的昼夜摄食节律与个体大小和性别无显著相关性。小规格雌、雄群体均具有最大的日摄食量,且雌性群体的日摄食量随着个体大小的增加显著降低。15 ℃条件下,大鳞副泥鳅群体的日摄食量占群体总体质量的2.2%~4.8%,个体的日摄食量约占个体体质量的3.0%。建议大鳞副泥鳅的投饲频率为一天2次,以体质量的3%~5%进行饲料投喂,最佳投饵驯化时间集中在6:00—7:00和18:00—19:00。     

Hematological Parameters and Blood Cell Morphology of Male and Female Schizothorax (Racoma) davidi (Sauvage)

The aim of this study was to obtain a basic knowledge of the hematology of Schizothorax (Racoma) davidi (Sauvage). The results showed that red blood cell and white blood cell counts as well as mean cellular hemoglobin concentration were significantly higher in females than in males, while mean cell volumes were significantly lower in females than in males. No significant differences were seen in other hematological parameters in relation with gender. Under light microscope, erythrocytes (mature and immature), leukocytes (neutrophils, lymphocytes, and monocytes), and thrombocytes were distinguished. Mature erythrocytes were ellipsoidal cells with oval, central nucleus. Neutrophils contained banded or segmented nuclei. Small lymphocytes had a very thin rim of basophilic cytoplasm. Monocytes often possessed vacuoles in their cytoplasm. Thrombocytes existed in several forms with relatively large nucleus surrounded by a pink‐stained or hyaline cytoplasm. The percentage of the different leukocytes revealed predominance of neutrophils and small lymphocytes, followed by monocytes and large lymphocytes. The neutrophil percentages of each stage showed that metamyelocyte accounted for the most, followed by banded neutrophils. This investigation could be used for evaluation of the health status of cultured male and female S. davidi and will provide early detection of clinical pathology.     

Nuclear transfer in loach (Paramisgurnus dabryanus Sauvage) by cell-to-cell electrofusion

To study nuclear transfer in the loach (Paramisgurnus dabryanus Sauvage), blastula and gastrula cells were fused with UV-inactivated oocytes by cell-to-cell electrofusion. To facilitate nuclear transfer, blastula and gastrula cells were cultured or incubated at 4 °C in different solutions. TC-199 medium supplemented with 20% calf serum was the best culture solution, and effectively retained the totipotence of blastula or gastrula cells for up to 10 days. It was found that gastrula cells incubated at 4 °C had the same totipotence as blastula cells. The optimal UV dosage for inactivation of the oocyte chromatin was 180–240 mJ cm⁻². Electrofusion was carried out in a cone-shaped fusion chamber, which permitted the recipient oocyte and the donor blastula cell to contact one another. The electrofusion procedure resulted in a 10% success rate of normal-appearing fish. Genetic analysis indicated that the nuclear material originated from the donor cell (blastomere) and the oocyte pronucleus did not take part in development.     

达氏鲟精巢细胞消化分离和超低温冷冻保存

通过研究两种酶对精巢细胞的消化效果,探究抗冻剂、降温程序、糖类和卵磷脂对达氏鲟(Acipenser dabryanus)精巢细胞冻存效果的影响,获得消化冻存后高存活率的细胞。实验使用0.25%胰蛋白酶和2 mg/m L胶原酶H+500 U/m L中性酶II的组合酶对达氏鲟精巢细胞进行消化,获得不同消化时间内的活细胞数量。另外,还研究了冷冻稀释液中分别添加10%二甲基亚砜(DMSO)、乙二醇(EG)、甲醇(MET)作为抗冻剂,采用-1℃/min慢速降温以及直接投入液氮中的快速降温方法冻存,冷冻稀释液采用D-海藻糖或同浓度D-蔗糖,以及添加5%、8%、11%卵磷脂对冻存效果的影响。结果显示:两种消化酶在同一时间消化所得的活细胞数和活细胞率无显著差异,并且都在3 h获得最多活细胞。慢速降温的冻存效果极显著地好于快速降温(P=0.01)。不同抗冻剂的保存效果差异显著,复苏后细胞相对存活率EG(51.70%±5.24%)MET(45.09%±3.15%)DMSO(40.18%±3.90%)。不同糖对达氏鲟精巢细胞冻存效果无显著影响;不同浓度的卵磷脂冻存效果有极显著差异。含8%卵磷脂的冻存液对细胞的冻存效果最好,细胞存活率可达(93.55±2.56)%,培养10 d后细胞数目为0 d时的3.19倍。     

Effect of Lactobacillus plantarum and Jerusalem artichoke (Helianthus tuberosus) on growth performance,immunity and disease resistance of Pangasius catfish (Pangasius bocourti,Sauvage 1880)

This study evaluated the effects of Jerusalem artichoke (JA) and Lactobacillus plantarum singly or combined on growth, immunity and disease resistance of Pangasius bocourti. In the first experiment, different concentrations of JA (0, 5, 10, 20, 40, 80 and 160 g kg^?1) were administered to determine an optimal concentration on growth of P. bocourti. In the second experiment, the optimal concentration of JA (5 g kg^?1) was combined with 10⁸ cfu g^?1 L. plantarum. In the final experiment, five randomly selected fish from the second experiment were challenged with Aeromonas hydrophila. Treatments for second and third experiments were 0 g kg^?1 JA (Diet 1), 5 g kg^?1 JA (Diet 2), 10⁸ cfu g^?1 L. plantarum (Diet 3) and 5 g kg^?1 JA + 10⁸ cfu g^?1 L. plantarum (Diet 4). Fish fed 5 g kg^?1 JA or 10⁸ cfu g^?1 of L. plantarum significantly improved specific growth rate (SGR), feed conversion ratio (FCR), serum lysozyme activity and postchallenge survival rate (PCSR). Dietary in the combination of JA and L. plantarum showed significantly enhanced SGR, FCR, serum lysozyme, phagocytosis, respiratory burst activities and PCSR compared with control and individual applications. Dietary JA and L. plantarum significantly stimulated growth, immunity and disease resistance of P. bocourti.     

外源核苷酸对斑点叉尾(鱼回)生长及肠道组织发育的影响

以斑点叉尾(鱼回)幼鱼为试验对象,研究不同水平外源核苷酸对其生长和肠道发育的影响.在基础饲料中分别添加0.0%(对照组)、0.02%、0.05%和0.1%的外源核苷酸.结果表明,在斑点叉尾(鱼回)幼鱼日粮中添加外源核苷酸,促生长作用不显著(P>0.05),但可显著促进肠道组织的发育(P<0.05).     

Spermiation of paddlefish (Polyodon spathula,Acipenseriformes) stimulated with injection of LHRH analogue and carp pituitary powder

The potential of carp pituitary powder (CPP) at one dose, or the luteinizing hormone-releasing hormone (LH-RH) analogue, des–Gly¹⁰,(d–Ala⁶)–LH-RH–ethylamide, at three different doses to stimulate spermiation in paddlefish (Polyodon spathula) was tested. Single injections of the LH-RH analogue at 0.2, 0.1, or 0.05 mg·kg^–1 increased the number of spermatozoa per kilogram of body weight (kg^–1 b.w.) by 4.7, 3.4, and 3.4 times respectively compared to control, but the number of spermatozoa per kilogram of body weight decreased with CPP (4 mg·kg^–1) by 1.7 times compared to the control. The LH-RH analogue prolonged active spermiation, with numbers of spermatozoa ranging from 7.69 to 1.19 × 10⁹ kg^–1 b.w. up to 96 h after treatment. Analysis of variance showed significant influence of experimental groups on volume of sperm per male and per kilogram of body weight, and the total number of spermatozoa per kilogram of body weight, but insignificant influence on the total number of spermatozoa per male. The percentage of motile spermatozoa was not different between experimental groups for sperm collection at different times after injection. A very high positive correlation (r = 0.93) was obtained between sperm concentration and sperm transmittance measured with a spectrophotometer. This relationship was described with the following linear regression: sperm concentration (× 10⁹ mL^–1) = 1.3244 X^–0.9969, where X is the percentage of sperm transmittance.     

Comparison of starch utilization in fingerlings of two Asian catfishes from the Mekong River (Pangasius bocourti Sauvage, 1880, Pangasius hypophthalmus Sauvage, 1878)

Five diets were formulated to provide an isoproteic daily dietary intake of 15 g kg^?1 day^?1, while maintaining daily starch intake ranging from 0 to 40 g kg^?1 day^?1. The 4‐week experiments started with initial mean weights of 4.7 and 4.4 g for Pangasius bocourti and P. hypophthalmus, respectively. The results clearly show the protein sparing action of starch in both species. Best protein retention was obtained with starch intake of 20 and 10 g kg^?1 day^?1 for P. bocourti and P. hypophthalmus, respectively, which was equivalent to 40 and 20% starch in the feed. Pangasius bocourti and P. hypophthalmus fingerlings attained maximal growth with starch intake of 30 and 10 g kg^?1 day^?1, equivalent to 60 and 20% starch in the feed. It was noted that body lipid accumulation was much higher in P. bocourti than in P. hypophthalmus, and that excess dietary starch intake depressed diet digestibility and growth.     

The Effects of Exogenous Bovine Growth Hormone and Placental Lactogen on Juvenile Striped Bass Morone saxatilis Feed and Growth Efficiency

Striped bass Morone saxatilis under normal pond-reared conditions usually requires 14–18 mo to grow out to market weight. This includes a winter of comparatively slower growth which could possibly be overcome through hormonally induced acceleration of growth. Thus, the objective of this experiment was to test the hypothesis that bovine growth hormone and bovine placental lactogen increase growth in striped bass in a dose dependent manner. A secondary objective was to determine the effect of these hormones on body composition. Variables tested were weight gain, length gain, condition factor, feed conversion, and body composition. Fish injected biweekly with 10 μg/g body weight, bovine placental lactogen ( P < 0.09) or bovine growth hormone ( P < 0.11) increased body weight. Fish injected with 1 or 10 μg/g body weight bovine placental lactogen ( P < 0.10) or bovine growth hormone ( P < 0.10) increased body length during the 12-wk study period. Proximate analysis indicated that bovine growth hormone increased protein content (10 μg/g; P < 0.10) and decreased fat content (1 and 10 μg/g; P < 0.10). No significant differences occurred in feed conversions. We conclude that bovine growth hormone and bovine placental lactogen are effective in increasing body growth in striped bass.     

Fertilization efficiency of cryopreserved sperm from striped catfish, Pangasius hypophthalmus (Sauvage)

The fertilization efficiency of cryopreserved sperm was compared with fresh sperm from striped catfish, Pangasius hypophthalmus . Of the two sets of experiments carried out, the first compared four sperm doses using fresh sperm and fresh eggs. The second experiment compared six concentrations of cryopreserved sperm ranging from 6.94 × 10⁷ to 6.94 × 10¹⁰ to fertilize 100 eggs per batch. Fertilization, hatch and survival rates were compared between cryopreserved and fresh sperm. The highest fertilization rate (53.75±1.62%) was achieved with a sperm dose of 6.94 × 10⁸. Increasing the sperm dose to 3.47 × 10⁹ did not increase the fertilization rate, indicating that the optimum sperm:egg ratio lies between 6.94 × 10⁶ and 3.47 × 10⁷ sperm per egg. Both highest (6.94 × 10¹⁰) and the lowest (6.94 × 10⁷) sperm doses resulted in lower fertilization rates (2.04% and 16.90% respectively). No significant differences were found among four fresh sperm doses compared. Mean hatch and survival rates resulting from fresh and cryopreserved sperm were similar. The experiment shows that while only 1.89 × 10⁶ fresh spermatozoa was required to fertilize a fresh egg, 6.94 × 10⁶ (or 3.67 times more) cryopreserved sperm was required to achieve the same level of fertilization. This provides important information for making decision to cryopreserve sperm for commercial and/or conservation purposes.     

外源核苷酸对异育银鲫幼鱼非特异性免疫的影响

以异育银鲫幼鱼为试验对象,研究不同水平外源核苷酸对其非特异性免疫的影响.试验分为4组,在基础饲料中分别添加0.0%(空白组)、0.02%、0.05%和0.10%的外源核苷酸,每组设3个的重复,每个重复40尾鱼.试验鱼体初始体重为1.03 g左右,每天饱食投喂2次,试验时间70 d.试验结束分别测定了添加0.0%、0.02%、0.05%和0.10%外源核苷酸的各组非特异性免疫指标.头肾指数分别为0.30%±0.01%、0.41%±0.01%、0.51%±0.02%和0.47%±0.01%;脾脏指数分别为0.23%±0.01%、0.31%±0.02%、0.37%±0.01%和0.37%±0.03%;白细胞数目分别为(4.74±0.71)×1011个/mL、(6.32±0.18)×1011个/mL、(6.56±0.12)×1011个/mL和(6.74±0.32)×1011个/mL;吞噬百分比分别为76.67%±2.03%、85.67%±1.33%、83.67%±3.48%和79.00%±2.00%;吞噬指数分别为(2.58±0.34)、(2.99±0.11)、(3.93±0.11)和(3.21±0.17);溶菌酶活力分别为(0.33±0.01)、(0.42±0.01)、(0.39±0.01)和(0.38±0.01).结果表明,在异育银鲫幼鱼日粮中添加0.05%外源核苷酸能够显著提高其机体的非特异性免疫功能(P＜0.05).     

Establishing bacterial infectivity models in striped Catfish Pangasianodon hypophthalmus (Sauvage) with Edwardsiella ictaluri

A bacterial infectivity challenge model of Edwardsiella ictaluri in striped catfish was developed. All experiments were conducted using a bacterial isolate of E. ictaluri that had been recovered during a natural outbreak of bacillary necrosis of Pangasianodon (BNP) in farmed striped catfish Pangasianodon hypophthalmus in Vietnam. Time of immersion in 10⁷ CFU.ml⁻¹ had a significant effect on mortality. The immersion bacterial dose of 10⁷ CFU/ml for 30 s resulted in a cumulative percentage mortality of 63%. Three to four days post-bacterial challenge, fish showed gross clinical signs of natural BNP and E. ictaluri was recovered and identified from these fish. Moreover, a cohabitation challenge was evaluated as an alternative challenge method, although the mortalities among the infected fish were lower at around 15%–40%. This study confirmed the horizontal transmission of E. ictaluri in striped catfish and elucidated that cohabitation challenge could be used in reproducing the disease under controlled conditions.