Efficacy of 17 α‐methyl testosterone and letrozole on sex reversal of protogynous grouper,Epinephelus tauvina (Forskal, 1775) and spawning performance of sex‐reversed males

This study aimed to test the efficacy of 17 α‐methyl testosterone (17 α‐MT) alone and in combination with letrozole, an aromatase inhibitor, for the induction of sex reversal in protogynous greasy grouper, Epinephelus tauvina. Further, the long‐lasting effects of these treatments and spawning performance of sex‐reversed males were also investigated. Greasy grouper with oocytes in the perinucleolus stage were implanted with 5 mg 17 α‐MT kg^?1 body weight (T₁), 5 mg 17 α‐MT and 0.2 mg letrozole kg^?1 body weight (T₂) and 5 mg 17 α‐MT with 0.4 mg letrozole kg^?1 body weight (T₃) and no androgens/enzyme inhibitor implanted (C). The 17 α‐MT alone and in combination of letrozole‐induced sex reversal in greasy grouper, whereas untreated control fish (C) showed normal ovarian development. However, T₂ and T₃ group showed 100% sex reversal and completion of spermatogenesis up to functional male phase in 2 and 3 months, respectively, whereas T₁ group resulted in only 66.67% functional male with motile spermatozoa after 4 months. Sex‐reversed males successfully fertilized the eggs during induced spawning. There were significant differences on fertilization and hatching rates between T₂ group (79.00 ± 4.36%; 77.67 ± 2.87%, respectively) and T₁ group (57.67 ± 3.17%; 63.87 ± 2.91%, respectively). The result suggested that 17 α‐MT (5.0 mg kg^?1 BW) in combination with letrozole (0.2 mg kg^?1 BW) has the potential to produce 100% sex‐reversed male in short period in greasy grouper, which might greatly help in seed production of greasy grouper.     

石斑鱼类养殖技术体系的创建和石斑鱼养殖产业持续发展的思考

石斑鱼是驰名世界的名贵海产鱼类,是中国南方沿海广东、海南、福建、广西等海水养殖业的主要对象,经济价值巨大。中国石斑鱼类养殖兴起于20世纪80、90年代;进入21世纪,中山大学的研究团队围绕石斑鱼苗种繁育和养殖产业化的理论和技术进行了一系列研究,对其生殖调控、生长发育、营养需求、病害防治、种质分析、养殖技术和养殖模式等方面进行集成创新,建立苗种繁育和成鱼养殖各个环节的技术规范和管理措施,实现了石斑鱼苗种规模化繁育和自养自足,从而带动石斑鱼养殖产业发展到相当的规模。石斑鱼类养殖产业化持续健康发展的总体思路是：以建设资源高效利用、改善生态环境、产品优质安全的现代渔业基本内涵为目标,改革和提升传统养殖技术和养殖模式,转变产业发展方式,不断优化和提升产业链上的各项技术,包括：（1）优化苗种人工繁育技术,提高苗种成活率和苗种质量;（2）培育抗逆、抗病、优质、高产的优良品种;（3）配制高效、优质的人工配合饲料;（4）建立病害检测和防控新技术和研制主要疾病的特异性疫苗;（5）建立高效、低碳、节能的养殖技术,因地制宜地采用先进的养殖模式;（6）加强石斑鱼类的生物学基础研究以及生物科学和工程技术的交叉融合、协同发展,为石斑鱼养殖产业的技术创新和集约化与工程化奠定基础。     

海南“石斑鱼苗网”渔具属性辨析

对在2004年7~8月间南海区小型海洋渔具渔法调查中发现的“石斑鱼苗网”的渔具结构、属性、性能和管理进行了分析讨论,研究认为“石斑鱼苗网”属于敷网类,网团型,延绳式。对石斑鱼苗有良好的诱捕效果,从保护资源的角度出发,应对其实施规范管理。     

Effects of dietary thiamin on the physiological status of the grouper <Emphasis Type="Italic">Epinephelus coioides</Emphasis>

This study was conducted to evaluate the effects of dietary thiamin on the physiological status of the juvenile grouper, Epinephelus coioides. Graded levels of thiamin (0.08, 0.50, 2.12, 3.15, 4.63, 12.37 mg thiamin kg⁻¹ diet) were fed to grouper juveniles (mean weight: 16.97 ± 0.14 g) for 10 weeks. Although fish fed the thiamin-deficient (TD) diet showed no obvious symptoms of thiamin deficiency or increased mortality, those fed the lowest doses of thiamin (0.08 and 0.50 mg thiamin kg⁻¹ diet) had significantly decreased transketolase activity in the liver. In addition, the level of liver thiobarbituric acid reactive substances in fish fed the TD diet was 33–67% higher than that in fish with the thiamin-supplemented diet. There were no significant differences in superoxide dismutase activity between the different groups of fish.     

Immune responses of orange-spotted grouper,Epinephelus coioides,against virus-like particles of betanodavirus produced in Escherichia coli

Betanodaviruses are the causative agents of viral nervous necrosis (VNN), a serious disease of cultured marine fish worldwide. Virus-like particles (VLPs) are one of the good novel vaccine candidates to control this disease. Until now, betanodavirus vaccine studies mainly focused on the humoral immune response and mortality after virus challenge. However, little is known about the activation of genes responsible for cellular and innate immunity by vaccines. In the present study, VLPs of orange-spotted grouper nervous necrosis virus (OGNNV) were produced in prokaryotes and their ability to enter Asian sea bass cells was the same as native virus, suggesting that they possess a similar structure to OGNNV. VLPs immunogenicity was then determined by intramuscularly vaccinating Epinephelus coioides at different concentrations (1.5 or 15 μg g^?1 fish body weight, FBW) and immunizing frequencies (administration once, twice and thrice). A single vaccination with the dosage of 1.5 μg g^?1 FBW is enough to provoke high titer antibodies (average 3 fold higher than that of negative control) with strong neutralizing antibody titer as early as 1 week post immunization. Furthermore, quantitative PCR analysis revealed that eleven genes associated with humoral, cellular and innate immunities were up-regulated in the liver, spleen and head kidney at 12 h post immunization, correlating with the early antibody response. In conclusion, we demonstrated that VLP vaccination induced humoral immune responses and activated genes associated with cellular and innate immunity against betanodavirus infection in orange-spotted grouper.     

用微卫星标记研究南海四种石斑鱼的遗传变异与种间关系(英文)

[目的]从DNA分子水平揭示我国南海的四种重要经济海水鱼类点带石斑鱼(Epinephelusmalabaricus)、斜带石斑鱼(E.coioides)、鲑点石斑鱼(E.fario)、蜂巢石斑鱼(E.merra)的遗传群体变异水平与种间关系。[方法]本研究利用从GenBank中搜索到的21对微卫星引物,对点带石斑鱼、斜带石斑鱼、鲑点石斑鱼及蜂巢石斑鱼的等位基因数A、有效等位基因数Ne、多态信息含量PIC、观察遗传杂合度Ho、期望遗传杂合度He等遗传参数进行检测,并采用UPGMA类聚分析法探讨这四种石斑鱼的种间关系。[结果]点带石斑鱼群体的A、Ne、PIC、Ho和He等遗传参数的平均值分别为4.38±1.60、3.69±0.86、0.69±0.08、0.67±0.08、0.72±0.06,斜带石斑鱼群体的为3.88±1.13、3.55±1.04、0.66±0.10、0.68±0.21、0.70±0.08,鲑点石斑鱼群体的为6.00±1.07、4.68±0.65、0.78±0.03、0.73±0.25、0.79±0.03,蜂巢石斑鱼群体的为5.50±1.07、4.58±0.80、0.76±0.05、0.75±0.18、0.78±0.04。[结论]综合比较上述数值可知,四个石斑鱼群体的遗传变异程度依次为鲑点石斑鱼群体>蜂巢石斑鱼群体>点带石斑鱼>斜带石斑鱼群体;与其他海水鱼类的遗传变异水平比较,四种石斑鱼的群体遗传变异水平都较高,说明我国这些石斑鱼种类的遗传种质资源尚好。另外,遗传距离与类聚分析表明,点带石斑鱼与斜带石斑鱼遗传距离最近,而斜带石斑鱼与蜂巢石斑鱼的亲缘关系最远。     

Evaluation of the Pathogenicity and Immune Response of Nervous Necrosis Virus Isolated in Vietnam

Eight strains of nervous necrosis virus （NNV） isolated in Vietnam were used to detect the pathogenicity and immune response in sea bass （SB）. All strains induced cytopathic effect in SB cell line, complete destruction of monolayer of cells appeared after seven days post infection （dpi）. Virus titer was different for each strain, TCIDso ranged from 102.7 to 1069, and LDs0 from 1015 to 1075. Five NNV strains named QN 02, QN 05, QN 07, ND 11 and KH 05 had higher virulence than the other three, the first causing 100% mortality in experimental fish 3-5 dpi. NNV KH 05 had the highest antigenic similarity, and it was inactivated completely by 0.2% formalin, 0.002 mol/L binary ethylenimine （BEI） and 0.1% beta-propiolactone. The neutralization antibody titer obtained in fish of groups immunized by BEI 0.002 M and beta-propiolactone 0.1% inactivated virus was four to eight times higher than that of the group treated with the formalin inactivated virus. The antibody titer in fish immunized with beta-propiolactone inactivated virus was more persistent. The efficacy of vaccines developed from beta-propiolactone inactivated virus and aluminium hydroxide （AH） or aluminum phosphate （AP） was observed by intramuscularly immunizing Epinephelusfuscoguttatus size 1.5 cm. Neutralizing antibodies appeared in vaccinated fish on 10th day post-immunization （dpi） at a dilution of 1：16; 1：32 and highest levels were reached on 30-45 dpi, at dilutions of 1：256 and 1：512, after treatment with AH and AP vaccine, respectively. The relative percent of survival （RPS） of vaccine at 30 dpi was highest with challenge doses 0.2-1 × 10^6.8 TCIDs0, the RPS varied from 80%-83.3% in both groups of AH and AP immunization. This result provides the basis for developing a vaccine against NNA disease.     

虎龙杂交斑养殖群体遗传多样性与遗传结构的微卫星分析

遗传多样性下降被认为是水产生物经济性状降低的常见原因。虎龙杂交斑作为养殖规模最大的石斑鱼品种,近年来病害增多。为了探究是否是由于群体遗传多样性的改变而影响虎龙杂交斑的养殖性状,本研究选用27个EST-SSRs标记,对苗种主产区6个虎龙杂交斑群体进行了遗传多样性和群体遗传结构分析。结果显示,6个群体其平均等位基因数（N_a）为6.666 7～8.555 6,平均有效等位基因数（N_e）为3.511 6～3.959 0,平均观测杂合度（H_o）为0.764 6～0.794 3,平均期望杂合度（H_e）为0.709 1～0.750 4,平均多态性信息含量（PIC）为0.662 2～0.706 4。分子方差分析（AMOVA）结果显示,5.70%的遗传变异来自群体间,0.00%来自群体内个体间,94.3%来自所有个体间。主坐标分析（PCo A）也显示差异主要来源于个体。群体间遗传分化指数(F_st)、遗传距离及聚类分析结果显示,HB和XC群体先聚为一支,再与翁田（WT）群体聚为一支,感城（GC）和...     

石斑鱼虹彩病毒病发生风险评估模型的建立和验证

为评估石斑鱼养殖过程虹彩病毒病发生的风险,实验结合Delphi法和层次分析法(analytic hierarchy process, AHP)构建新加坡石斑鱼虹彩病毒(SGIV)发生的风险评估模型。风险因素评估指标体系包括1个目标层(石斑鱼虹彩病毒病发生风险),5个准则层(水质、石斑鱼健康状况、饲养管理、养殖模式和养殖环境)和20个指标层风险因素(水温、pH、溶解氧、氨氮、病毒感染、细菌感染等)。准则层风险因素权重值集合为W={0.129 8,0.367 2,0.173 3,0.032 7,0.297 1},指标层风险因素中病毒感染(0.248 5)、邻近区域发病情况(0.138 4)、水温(0.112 3)和养殖密度(0.105 0)等权重值较高,推测它们是影响石斑鱼虹彩病毒病暴发的高风险因素。进一步实验室模拟感染实验结果表明,养殖密度、病毒感染剂量及养殖温度均显著影响鱼体内新加坡石斑鱼虹彩病毒(SGIV)的复制及实验鱼死亡率。上述结论和评估模型中推定的高风险因素相吻合,说明该模型可用于石斑鱼养殖过程中虹彩病毒病发生的风险评估。