七带石斑鱼仔鱼摄食习性的观察

在人工育苗条件下,采用轮虫与牡蛎受精卵两种开口饵料,3种不同的投喂方式：开口单喂轮虫(A)、开口单喂牡蛎受精卵(B)、开口将牡蛎受精卵和轮虫混合投喂(C),分析对比不同饵料对仔鱼开口效果。通过观察5日龄仔鱼日摄食与4~20日龄仔鱼消化道饱满度的变化,探讨仔鱼摄食习性及光照与摄食的关系。结果发现,七带石斑鱼孵化开口后,投喂3d牡蛎受精卵者比直接投喂轮虫对仔鱼生长有明显的效果;10日龄前,B组在全长、肛前距、口裂宽方面都大于A、C两组,B组全长的特定生长率为4.969%,分别是A、C两组的2.5与2.4倍,生长速度最快;5日龄仔鱼在09∶00~10∶00与13∶00~14∶00时间段,有两个明显的摄食高峰期,光照强度分别为720~1 018 lx与865~923 lx;4日龄仔鱼其饱食率较低,只有40%达到了3级摄食量,4~10日龄的仔鱼饱食率波动幅度较大,10日龄后开始逐渐上升,13日龄后基本可以达到100%。     

七带石斑鱼人工繁育技术研究进展

七带石斑鱼(Epinephelus septemfasciatus)具有经济价值高、生长速度快和低温耐受性强等特点,是东北亚温带海域理想的网箱养殖品种之一,已成为中、日、韩三国海水鱼类繁育研究的热点。由于存在雄鱼性成熟年龄高、人工驯养条件下难以实现自然产卵和初孵仔鱼弱小等难题,其稳定和高效的种苗规模化繁育技术尚未完全建立起来。本文综述了七带石斑鱼人工繁育技术研究现状,重点总结日本近20年在亲鱼繁殖、仔稚鱼培育和苗种中间培育等方面取得的研究进展,并就其发展前景进行分析,旨为中国七带石斑鱼苗种规模化繁育技术研发提供参考。     

Skeletal development and deformities in cultured larval and juvenile seven-band grouper, Epinephelus septemfasciatus (Thunberg)

The seven‐band grouper, Epinephelus septemfasciatus (Thunberg), is currently recognized as a potential new species for aquaculture in Japan. This study describes normal and abnormal skeletal development of the jaw and vertebrae in cultured larvae and early juveniles of E. septemfasciatus. The ontogenetic stages at which skeletal deformities of jaw and vertebra developed were also described for this species. Osteological observations were made using a clearing and staining method for larvae and soft X‐ray photographs for juveniles. A high incidence of skeletal deformities was observed in the jaws and vertebral column during the larval and juvenile stages. Most of the jaw deformities were explained by an abnormal maxilla curvature. Jaw deformities were visually evident from flexion stage after ossification of the deformed elements. Deformities in the vertebral column (mostly lordosis) were observed from the post‐larval stage and became more evident as growth proceeded. The lordosis generally occurred on the positions of the 8–11th vertebra. These types of deformities are compared with those of other species, and possible causative factors of the skeletal deformities are discussed.
     

Effect of temperature on survival,growth and malformation of cultured larvae and juveniles of the seven-band grouper Epinephelus septemfasciatus

The effects of rearing temperature (23–29 °C) during the larval and juvenile stage on survival, growth and skeletal malformations in the seven-band grouper Epinephelus septemfasciatus were investigated. The survival rate of juveniles 30–40 mm in total length emerging from eggs was higher at 25 and 26 °C (0.1–1.3 %) than at 23 °C or 27–29 °C (0.004–1.5 %). Growth (increase in total length) was accelerated at higher temperatures. The frequency of malformed individuals was lower at 25–27 °C (36.0–61.5 %) than at 23, 28 or 29 °C (65.3–76.9 %). Specific incidences of spinal curvature and centrum fusion or defects in juveniles were not related to rearing temperature. However, incidences of twisted or compressed vertebrae (6.5–64.0 %) were higher at higher temperatures, while the incidence of bifurcated neural spine was significantly higher at 23 °C (43.6–54.4 %) than at other temperatures (3.3–22.7 %). The incidence rate of spinal curvature (23.3 %) was significantly higher in juveniles with a deflated swim bladder, regardless of rearing temperature. The results of this study suggest that the optimum culture temperature for seven-band grouper is 25–26 °C, collectively considering the survival, growth and incidences of abnormalities. Our results also demonstrate the significance of identifying the conditions for swim bladder inflation to prevent spinal curvature in seven-band grouper.     

Improvement of the survival in the seven-band grouper Epinephelus septemfasciatus larvae by optimizing aeration and water inlet in the mass-scale rearing tank

The water flow in larval rearing tanks has been indicated to cause mass mortality of the seven-band grouper Epinephelus septemfasciatus larvae. Therefore, a new aerating method was tested in an actual scale intensive rearing tank (8.0 m in diameter, 1.87 m of water depth, 100 m³ of volume), in which an aerator was positioned at the center of the rearing tank surrounding cylindrical drain (1.2 m in diameter) to generate the flow field, and seven larval rearing trials were performed. The survival rate with the former aeration methods were compared, in which several aerators were located in the rearing tank. The survival rate at 10 days after hatching with the new aeration method (61.5±5.1%, n=7) was approximately three times higher than the former methods (21.2±13.7%, n=6). The flow environment of rearing tanks was also examined by quantifying the flow field, and the relationship between the flow field in the rearing tank, behavior of larvae and survival discussed. It was confirmed that the vertical circulating flow was observed in rearing tanks, and determined effectively the survival and the behavior of grouper larvae in patchiness.     

七带石斑鱼的形态和生物学性状

2010年在广东省饶平县柘林湾进行了七带石斑鱼（Epinephelus septemfasciatus）海水网箱人工养殖研究,观察和研究了人工养成的七带石斑鱼的形态特征和生物学性状。通过观察、测量、解剖,对114尾七带石斑鱼的外部形态、可量比性状和内部结构进行了研究。结果表明,七带石斑鱼体呈椭圆形,侧扁,体侧具7条褐色横带,第1条横带仅伸达主鳃盖骨上方边缘,第6条横带分成2条,体侧横带具有不规则的白色斑纹,各鳍边缘白色,背鳍鳍棘部褐色和白色横带相间,鳍条部具有由白色横带延伸而成的3个白斑。胃呈卜型,幽门盲囊发达;肠道长度为体长的0.7～1.4倍,与七带石斑鱼为肉食性鱼类的特征相一致。     

Effect of tank proportions on survival of seven-band grouper Epinephelus septemfasciatus (Thunberg) and devil stinger Inimicus japonicus (Cuvier) larvae

We examined the effects of rearing‐tank proportions on early survival, surface death and growth of the seven‐band grouper Epinephelus septemfasciatus (Thunberg) and the devil stinger Inimicus japonicus (Cuvier). Fertilized eggs were introduced into three differently shaped 100 L rearing tanks. The three tanks had different water surface areas, and included a shallow tank (S; 71 × 26 cm in diameter and depth, respectively), an intermediate tank (I; 57 × 39 cm) and a deep tank (D; 44 × 70 cm). Both species showed their highest survival rate and the lowest numbers of surface death in the D tank (P<0.05). There were no significant differences between fish reared in the three tank shapes in notochord length, total length, growth rate and dry weight. Rearing‐tank shape affected larval movement in the water column, with the duration of larval movement under the water surface being the shortest in the D tank. These results suggest that using a rearing tank of a suitable shape could significantly reduce the surface death of marine fish larvae.     

Molecular cloning of gonadotropin cDNA in sevenband grouper, Epinephelus septemfasciatus

The cDNA for glycoprotein hormone (GPH) α-subunit, β-subunits of the FSH (GtH I) and LH (GtH II) were isolated, cloned, and sequenced from sevenband grouper pituitary using RACE PCR. These results show that sevenband grouper has two different types of gonadotropins FSH (GtH I) and LH (GtH II), as other teleosts.     

七带石斑鱼染色体核型研究

采用PHA体内直接注射法制备了七带石斑鱼(Epinephelus septemfasciatus)头肾组织染色体标本并分析其核型。结果表明,七带石斑鱼二倍体染色体数为48,24对染色体均为端部着丝粒染色体,其染色体臂数(NF)为48,核型公式为:2n=48t,NF=48。未发现与性别相关的异型染色体,第24对染色体长度明显小于其他染色体。通过与其他22种石斑鱼染色体核型进行比较,发现七带石斑鱼具有石斑鱼属鱼类的原始核型特征,属于石斑鱼属鱼类的原始类群。     

Effect of temperature,hypoxia and disinfection with ozonated seawater during somitogenesis on muscular development of the trunk in larval seven‐band grouper,Epinephelus septemfasciatus (Thunberg)

The cause of abnormal musculature on both sides of the same myotome in artificially cultured seven‐band grouper Epinephelus septemfasciatus (Thunberg) larvae remains unclear. We evaluated the effect of three factors: changes in temperature (between 22 and 24°C), disinfection with ozonated seawater (0.3 mg TROs L^?1, 1 min), and hypoxia (22 or 14% saturation) during the early (20 h post‐fertilization, hpf) and mid‐ (24 hpf) somite stages on trunk muscular development in larval seven‐band grouper. The thermal regimes and disinfection with ozonated seawater had no effect on muscle development (P > 0.05, G‐test). In contrast, exposure to hypoxia during both the early and mid‐somite stages increased the incidence of abnormal musculature in the trunk (P < 0.01, G‐test). These larvae had abnormal musculature on both sides of the trunk. In addition, the number of abnormal myotomes differed among these larvae. Exposure in the early and mid‐somite stages resulted in the highest frequency of abnormalities in the 11th and 13th myotome respectively. Our results suggest that hypoxia during somitogenesis can lead to abnormal muscle development in the trunk of artificially cultured seven‐band grouper larva.     

盐度对七带石斑鱼胚胎发育和卵黄囊仔鱼生长的影响

观察比较了8个盐度梯度[10、15、20、25、30(自然海水)、35、40和45]下七带石斑鱼Epinephelus septem fasciatus受精卵的孵化周期、孵化率和畸形率及卵黄囊仔鱼的畸形率,并在上述条件下对七带石斑鱼进行了耐饥饿试验,测定其生存活力指数(SAI).结果显示,受精卵孵化的最适盐度范围为30～35,盐度高于35,孵化率随着盐度的升高而降低,仔鱼畸形率随之升高;盐度低于25,孵化率随盐度的降低而降低,仔鱼畸形率随之升高.仔鱼的最适生存盐度为30～35,此盐度下的仔鱼存活系数为24.7±0.36和23.27土3.01.     

Embryonic and larval development of honeycomb grouper Epinephelus merra Bloch

Spawning and successful rearing of larvae of honeycomb grouper Epinephelus merra Bloch 1793 upto juvenile stage was accomplished at the finfish hatchery of Mandapam Regional Centre of Central Marine Fisheries Research Institute during 2004. The fertilized eggs were free, spherical and buoyant with size ranging from 710 to 730 μm. Complete early embryonic development took place within 24–27 h and hatching occurred. The hatchlings measured 1.5 mm. Mouth opening (115 μm) appeared at 72 h when the larvae were 2.2 mm in size. Pectoral fin developed on the fifth day. Complete metamorphosis took place and by the 60th day the larvae transformed into juveniles (45 mm) and attained skin colouration and honeycomb pattern.     

Dry shipper cryopreservation of seven‐band grouper (Epinephelus septemfasciatus Thunberg) spermatozoa

This study examined the usage of a dry shipper for cryopreservation of Epinephelus septemfasciatus (Thunberg) spermatozoa. Milt was diluted 1:49 with 5% dimethyl sulfoxide plus 95% foetal bovine serum for cryopreservation. Computer‐assisted sperm analysis was used to analyse sperm motility, while fertilization and hatching trials were conducted to gauge the applicability of the cryopreservation method for aquaculture. We showed that cooling rates of the dry shipper were stable for 14 days and could be manipulated by the use of different sized freezing straws and use of a simple polystyrene foam container (5 × 5 × 12 cm and 1 cm thickness on all sides with the upper layer exposed). Dry shipper cryopreserved spermatozoa had significantly lower post‐thaw per cent motility and velocity than fresh sperm, but linearity of movement was unchanged. Fertilization and hatching rates were not significantly different at all tested sperm to egg ratios (3000:1–243000:1). The results indicated that 0.33 mL of milt when cryopreserved was sufficient to fertilize up to 450 g of oocytes. Application of this technology will help improve seed production in aquaculture and further develop breeding and genetics studies.     

Protective immunity of sevenband grouper, Epinephelus septemfasciatus Thunberg, against experimental viral nervous necrosis

This paper describes the protective immune responses of sevenband grouper, Epinephelus septemfasciatus Thunberg, immunized with live piscine nodavirus, the causative agent of viral nervous necrosis (VNN), or the Escherichia coli – expressed recombinant coat protein. Nodavirus-neutralizing antibodies were detected at titres ranging from 1:158 to 1:1257 in serum of sevenband grouper which survived intramuscular injection with the virus, by a cell culture assay system. The virus-neutralizing ability of immune serum was also confirmed by injecting virus previously treated with serum into fish. This indicates establishment of acquired immunity in survivors and thus explains why survivors from natural infection are resistant to recurrence of the disease. Young sevenband grouper were immunized twice by intramuscular injections with the recombinant coat protein. Immunized fish produced neutralizing antibodies at high titres for at least 110 days and showed significantly lower mortalities in virus challenge tests. These results suggest the potential for vaccination against VNN in sevenband grouper, which is susceptible to piscine nodavirus at all life-stages.     

Histopathological studies on viral nervous necrosis of sevenband grouper, Epinephelus septemfasciatus Thunberg, at the grow-out stage

Abstract Viral nervous necrosis caused by sevenband grouper nervous necrosis virus (SGNNV) has occurred in grow-out stages (0-3 years old) of sevenband grouper, Epinephelus septemfasciatus, since the 1980s. In the present study, based on histopathological features of the central nervous system (CNS) in naturally diseased fish, pernasal infection experiments using grow-out fish were performed and pernasal infection was established as a putative invasion route of SGNNV. The definite SGNNV-targeted cells were determined by histopathological studies including indirect fluorescent antibody test and electron microscopy. Nerve cells in the olfactory lobe were most extensively necrotized with vacuolation followed by infiltration of microglia and macrophages. Purkinje cells and Golgi cells were extensively infected in the cerebellum. Megalocells and small nerve cell nuclei were also infected in the preoptic area, thalamus, medulla oblongata and spinal cord. Only a few small nerve cells were infected in the olfactory bulb and optic tectum. The retina of some diseased fish displayed vacuolated bipolar cells of the inner nuclear layer and in the ganglion cell layer. These SGNNV-infected nerve cells displayed viroplasmic inclusions containing virions, vacuoles and myelin-like structures. Based on observed histopathological changes, the lesion of the CNS was characterized by encephalitis but not encephalopathy.     

Experimental evaluation of inorganic fertilization in larval giant grouper (Epinephelus lanceolatus Bloch) production

A major constraint in successful larviculture of groupers has been the small gape of the larvae and hence the requirement for small prey at first feeding. In this study, we examined how maintaining a phosphate concentration of 100 μg P L^?1 and an inorganic nitrogen (N) level of 700 μg N L^?1 via weekly fertilization with inorganic fertilizers affected phytoplankton, zooplankton and giant grouper larval survival in relation to a control group that was provided with rotifers immediately after larvae hatched. Unicellular algae, zooplankton within the size ranges of 10–50 μm and 50–100 μm and survival of giant grouper larvae were all significantly higher in the fertilized treatment compared with the control. Stomach analysis revealed that ciliates and flagellates were actively consumed by larval fish in the fertilized group, whereas few rotifers were consumed in the control. We conclude that the inorganic fertilization method provides high densities of suitable‐sized prey for larval groupers at the onset of exogenous feeding before they are able to consume larger, commercially available rotifers and copepods.     

Ontogeny of digestive and immune system organs of larval and juvenile kelp grouper Epinephelus bruneus reared in the laboratory

ABSTRACT:   A histological examination was made of the ontogenetic development of the digestive and immune systems of the larval and juvenile kelp grouper Epinephelus bruneus reared in the laboratory. The liver, gall bladder, pancreas and the demarcating region between the intestines and rectum were formed within 3 days post-hatch (dph). During the preflexion phase (within 16 dph), revolution of the intestine concluded, and pharyngeal teeth and the mucous cells of the esophagus were differentiated. In the transitional period to the juvenile stage (25 dph), the blind sac of the stomach, gastric glands and pyloric caeca began to form. From the viewpoint of the differentiation phase of the adult-type digestive system, the kelp grouper is similar to Heterosomata, hitherto reported. The primordial thymus, kidney and spleen were present at 12, 1 and 6 dph, and the small lymphocytes in these lymphoid organs appeared at 21, 30 and 33 dph, respectively. The developmental sequence of the lymphoid organs and the appearance ages of the lymphoid organs and small lymphocytes in the lymphoid organs in the kelp grouper are similar to those of other marine fish previously reported, except for the Pacific bluefin tuna Thunnus orientalis .     

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.