Protection conferred against viral nervous necrosis by simultaneous inoculation of aquabirnavirus and inactivated betanodavirus in the sevenband grouper, Epinephelus septemfasciatus (Thunberg)

An aquabirnavirus (ABV) and a formalin-inactivated betanodavirus [redspotted grouper nervous necrosis virus (RGNNV)] were investigated for their potential to prevent RGNNV-induced viral nervous necrosis (VNN) in the sevenband grouper, Epinephelus septemfasciatus (Thunberg). Three groups of fish were injected intramuscularly with ABV, intraperitoneally with inactivated RGNNV (iRGNNV) or with both ABV and iRGNNV. At 3, 7, 14, 21 and 28 days post-injection (p.i.), fish were challenged by intramuscular injection of RGNNV. Control fish, which received neither ABV nor iRGNNV, showed high mortalities in all RGNNV challenges. Fish that received only ABV exhibited relative percent survival (RPS) of >60 against RGNNV challenges at 3, 7, 14 and 21 days p.i., but not at 28 days p.i., while fish that received only iRGNNV showed significantly higher protection against RGNNV challenges only at 21 and 28 days p.i. In contrast, fish that received both ABV and iRGNNV showed 60 or higher RPS against all RGNNV challenges. Fish inoculated with iRGNNV with or without ABV exhibited similar high titres of neutralizing antibodies to RGNNV at 14, 21 and 28 days p.i. These results indicate that combined inoculation with iRGNNV and ABV conferred both rapid non-specific and delayed specific protection against VNN.     

赤点石斑鱼与鞍带石斑鱼杂交子一代肌肉营养成分分析与评价

为分析赤点石斑鱼与鞍带石斑鱼杂交子一代的营养组成,参照国家标准,测定了体质量(182.84±29.35) g杂交石斑鱼肌肉的常规营养成分、氨基酸和脂肪酸组成,并对肌肉营养价值进行了评定。试验结果显示,杂交石斑鱼肌肉水分、粗蛋白、粗脂肪和灰分的含量分别为(74.07±0.71)%、(21.52±0.78)%、(4.03±0.15)%和(1.29±0.07)%。肌肉鲜样中测定了17种氨基酸,总量为(19.88±0.15)%;必需氨基酸和鲜味氨基酸总量分别为(8.64±0.13)%和(7.64±0.16)%,必需氨基酸指数为85.19,必需氨基酸组成符合联合国粮农组织/世界卫生组织标准。肌肉鲜样中含有17种脂肪酸,饱和脂肪酸、单不饱和脂肪酸和多不饱和脂肪酸分别占肌肉脂肪酸总量的(27.63±1.15)%、(22.75±1.22)%和(32.59±1.90)%,其中二十二碳六烯酸和二十碳五烯酸占肌肉脂肪酸总量的(19.27±1.27)%。研究表明,赤点石斑鱼与鞍带石斑鱼杂交子一代具有较高营养价值,可作为新品种进行开发。     

福建南部养殖石斑鱼匹里虫(Pleistophora sp.)病观察

近年来，福建南部海区一些网箱养殖石斑鱼常见一种流行性孢子虫病，病鱼主要表现为游动失常、厌食、消瘦。外表无明显病理症状，部分病鱼腹部膨大变形，严重者可引起死亡。剖检可见腹腔、生殖腺、肝脏、结缔组织以及肌肉组织内散布或成串排列数个到数十个大小不一的黑色或银灰色孢囊。包囊内含大量孢子虫的子孢子，子孢子呈圆形或椭圆形，大小约4～5μm。根据形态学以及寄生特点确定该寄生虫是一种匹里虫。     

饥饿对斜带石斑鱼肝脏和肌肉脂肪酸组成的影响

在室外水泥池(4m×1.5m×1m)进行石斑鱼(62.3±0.6g)的饥饿实验,每池放养35尾,设3个重复,实验时间分别为0、1、2、3和4周。随着饥饿时间的延长,体重减轻,肥满度和肝体比显著降低;肌肉水分显著增加而蛋白质含量显著下降(P<0.05),脂肪含量在第1周下降后基本保持稳定。肝脏饱和脂肪酸在饥饿前3周基本保持不变,第4周显著下降。单不饱和脂肪酸在饥饿第1周保持不变,然后呈下降趋势。多不饱和脂肪酸和高不饱和脂肪酸的总和随着饥饿时间的延长,含量显著增加。与肝脏相比,肌肉中脂肪酸变化程度较小。随着饥饿时间的延长,肌肉中140减少而205增加。饥饿使肌肉中DHA/EPA比率呈下降趋势,但肝脏DHA/EPA比率显著上升;肌肉和肝脏中DHA EPA的百分比都随饥饿时间的延长而显著上升。     

循环水养殖条件下鞍带石斑鱼生长特点研究

通过对鞍带石斑鱼(Epinephelus lanceolatus)为期5个月的养殖监测试验,研究循环水养殖条件下鞍带石斑鱼的生长特点。采用线性拟合、指数拟合、乘幂拟合的方法,分析了鞍带石斑鱼全长、体重生长与养殖时间,以及全长与体重的最佳拟合曲线。结果表明:鞍带石斑鱼全长生长与养殖时间以线性回归为佳(y=2.791x-15.716,R2=0.951 2);体重生长与养殖时间以指数回归最佳(y=2.432 3e0.32x,R2=0.996 5);全长与体重以乘幂回归为最佳(y=0.007 3x3.262 8,R2=0.945 3)。对鞍带石斑鱼全长与体重的幂函数关系分析,表明鞍带石斑鱼属正异速生长型(b=3.262 8),说明本研究条件下的循环水养殖模式适用于鞍带石斑鱼的养殖。     

鞍带石斑鱼仔稚（幼）鱼的发育和生长研究

对鞍带石斑鱼的仔稚幼鱼形态发育的各个阶段进行了观察与研究,详细描述从初孵仔鱼到幼鱼各个发育时期的形态特征和发育时间。根据卵黄囊的变化,长鳍棘的长出与收缩,鳞片和体色斑纹的出现,鞍带石斑鱼胚后发育可以划分为仔鱼期、稚鱼期、幼鱼期。仔鱼期又可分为卵黄囊期仔鱼和后期仔鱼。水温27～30℃,盐度27～31,pH值8.0～8.4的海水中培育,初孵仔鱼至孵化后2日龄为卵黄囊期仔鱼。2日龄仔鱼开口,3日龄至20日龄为后期仔鱼,22日龄至30日龄为稚鱼期,31日龄进入幼鱼期。鞍带石斑鱼胚后发育过程中最明显的变化是背鳍棘和腹鳍棘的生长和收缩,也是生产育苗当中比较关键的仔稚幼鱼变态过程。     

温度和盐度对鞍带石斑鱼受精卵发育及仔鱼成活率的影响

研究了在8个盐度梯度(18.3‰、19.7‰、21.2‰、23.5‰、25.6‰、27.1‰、29.5‰和31.7‰)下鞍带石斑鱼Epinephelus lanceol受精卵和仔鱼在水中的分布状态、出膜率和仔鱼存活率;在9个温度梯度(18℃、20℃、22℃、24℃、26℃、28℃、30℃、33℃和35℃)下,其受精卵孵化率、畸形率和仔鱼成活率。结果表明:(1)在水温为25~26℃,当盐度≤25.6‰时,受精卵表现为沉性;当盐度≥27.1‰时,受精卵表现为浮性。仔鱼的分布与受精卵有相似的特点,当盐度≤21.2‰时仔鱼分布在底层水体;当盐度≥27.1‰时,仔鱼集中分布在上层水体。(2)以孵化率和成活率为指标,鞍带石斑鱼孵化的适宜盐度范围为21.2‰~31.7‰,最适盐度范围为25.6‰~31.7‰。在盐度为31.7‰时仔鱼成活率最高达到75.9%。(3)鞍带石斑鱼胚胎发育的适温范围是26~30℃。28℃时受精卵孵化率、仔鱼成活率均最高,畸形率最低。     

Effects of dietary live or heat‐inactivated autochthonous Bacillus pumilus SE5 on growth performance,immune responses and immune gene expression in grouper Epinephelus coioides

To evaluate the possible dietary application of live and heat‐inactivated probiotic Bacillus pumilus SE5 in grouper Epinephelus coioides, juveniles (14.6 ± 0.2 g) were fed either a basal control diet (without probiotic) or the basal diet supplemented with 1.0 × 10⁸ CFU g^?1 live (T1) and heat‐inactivated B. pumilus SE5 (T2). The heat‐inactivated probiotic significantly improved the final weight, weight gain (WG) and specific growth rate (SGR) at day 60 and significantly decreased the feed conversion ratio (FCR) at day 30 and 60, while the viable probiotic significantly decreased the FCR at day 60 (P < 0.05). Phagocytic activity, serum complement C3 and IgM levels as well as SOD activity elevated significantly in fish fed the heat‐inactivated probiotic for 60 days (P < 0.05). Furthermore, the heat‐inactivated probiotic remarkably up‐regulated expression of TLR2 and pro‐inflammatory cytokines (IL‐8 and IL‐1β) in head kidney (P < 0.05), but the viable probiotic failed to do so. These results indicated that heat‐inactivated B. pumilus SE5 can effectively improve the growth performance and immune responses of E. coioides.     

斜带石斑鱼病原菌(哈维氏弧菌)的分离与鉴定

从患病斜带石斑鱼 (Epinepheluscoioides)肝脏组织分离到EcGY0 2 0 4 0 1菌株 ,经人工感染、回归感染实验证实为致病菌。通过API系统和菌体常规形态特征、培养特性和生理生化反应指标测定以及 16SrRNA测序分析等综合鉴定 ,Ec GY0 2 0 4 0 1菌株为弧菌属哈维氏弧菌 (Vibrioharveyi) ,其半致死剂量LD50 为 2 .7× 10 6CFU/g鱼体重。药敏试验结果表明 ,EcGY0 2 0 4 0 1菌株对利福平、四环素、喹诺酮类及头孢曲松等抗生素较为敏感     

斜带石斑鱼肝细胞分离及原代培养方法的建立

本研究以斜带石斑鱼肝细胞为实验对象,在不同培养条件下进行原代培养,旨在探讨稳定可靠的斜带石斑鱼肝细胞分离及原代培养方法。采用组织块分离法和胰蛋白酶(含EDTA)消化法分离肝细胞,并通过密度梯度离心法分离纯化肝细胞,细胞悬液于DMEM/F-12、M199和L-15培养液中培养;细胞活力及数量采用血球计数板计数,并通过MTT法测定细胞增殖率;同时,测定不同时间培养上清液中乳酸脱氢酶(LDH)活性、白蛋白(ALB)和尿素氮(BUN)的含量,以分析肝细胞生长状态。结果表明,组织块方法不适于斜带石斑鱼肝细胞的培养,未见细胞从组织块中迁出,而胰蛋白酶消化法获得良好稳定的培养效果,细胞产量达到1.6×108个/g肝重,活细胞数达到95%;L-15培养基细胞生长明显优于DMEM/F-12和M199培养基;启动原代培养的48~72 h阶段肝细胞生长代谢旺盛,培养上清液中LDH活性显著降低,ALB和BUN含量显著升高。结果显示,0.25%的胰蛋白酶常温消化法适合斜带石斑鱼肝细胞的分离,斜带石斑鱼肝细胞原代培养的最适培养基为L-15培养基,肝细胞在启动原代培养的48~72 h生长代谢旺盛。