Histopathological Observation of Lymphocystis Disease and Lymphocystis Disease Virus (LCDV) Detection in Cultured Diseased Sebastes schlegeli

Lymphocystis nodules occurring in the cultured sting fish Sebastes schlegeli were observed under light and electron microscope. Lymphocystis disease virus (LCDV) in the tissues of diseased fish was detected with indirect immunofluorescence test (IFAT). Results showed that lymphocystis cells had overly irregular nuclei, basophilic intracytoplasmic inclusion bodies with virions budding from the surface, and hyaline capsules outside the cell membrane. Numerous virus particles about 200 nm in diameter scat- tered in the cytoplasm, electron-dense particles 70-80 nm in diameter filled in perinuclear cisterna, and membrane-enveloped parti- cles with electron-dense core of 70-80 nm appeared around cellular nucleus. IFAT using monoclonal antibody against LCDV from Paralichthys olivaceus revealed that specific green fluorescence was present in the cytoplasm of lymphocystis cells, epithelium of stomach, gill lamellae, and muscular fibers under epidermis of S. schlegeli, just as that in the cytoplasm of lymphocystis cells of P. olivaceus, suggesting the presence of LCDV in these tissues.

Histological Observation of Germ Cell Development and Discovery of Spermatophores in Ovoviviparous Black Rockfish(Sebastes schlegeli Hilgendorf) in Reproductive Season

Black rockfish(Sebastes schlegeli) is an important species for culture; however, its reproductive characteristics have not been fully documented. In this study, we investigated the morphology and developmental process of germ cells in this ovoviviparous rockfish in reproductive season(October 2011–November 2012) with histological methods. We found that the gonad of mature fish showed notable seasonal changes in developmental characteristics and morphological structure. The sperm cells matured during a period lasting from October to December, significantly earlier than the oocytes did. A large number of spermatozoa and other cells occurred in testis at different developmental stages. Vitellogenesis in oocytes began in October, and gestation appeared in April next year. Spermatophores were discovered for the first time in Sebastes, which assembled in testis, main sperm duct, oviduct and genital tract, as well as ovarian cavity in October and April. These organs may serve either as production or hiding places for spermatophores and spermatozoa which were stored and transported in form of spermatophores. Testicular degeneration started from the distal part of testis in April, with spermatophores assembled in degenerating testis and waiting for transportation. The copulation probably lasted for a long period, during which the spermatozoa were discharged in batches as spermatophores. These spermatophores were coated with sticky materials secreted from the interstitial areas of testis and the main sperm duct, then transported into ovary.

The application of stereo-video technology for the assessment on population change of black rockfish Sebastes schlegeli in a vessel reef area in Haizhou Bay,China

The assessment of population structure and abundance of fish assemblages associated with artificial reefs(ARs) is an important aspect of AR management.In the present study,we used a DiveOperated Stereo Video(stereo-DOV) technique to assess the population structure and abundance of Sebastes schlegeli associated with two metallic,and three wooden,vessel reefs in Haizhou Bay during 2012 and2013.The study used video systems to obtain length measurements and estimates of abundance.The size composition of S.schlegeli differed among reefs and individuals around vessel reefs were all adults,with total lengths(TL) of >20 cm.Juvenile fish were encountered by divers in a rocky area near the island away from the vessel reefs.The largest individual S.schlegeli(with the highest TL) among five reefs were found around a metallic vessel reef in both 2012 and 2013.TL of S.schlegeli from all reefs increased by an average of 3.2 cm(P<0.05) from 2012 to 2013,with an estimated mean weight increase of 250.4 g(P<0.05).The video survey also indicated a decrease in the biomass of schools near two metallic vessels between the years.Stereo-video technology was found to be suitable for rockfish surveys around the reefs.

A Microsatellite Genetic Linkage Map of Black Rockfish(Sebastes schlegeli)

Ovoviviparous black rockfish(Sebastes schlegeli) is an important marine fish species for aquaculture and fisheries in China. Genetic information of this species is scarce because of the lack of microsatellite markers. In this study, a large number of microsatellite markers of black rockfish were isolated by constructing microsatellite-enriched libraries. Female- and male-specific genetic linkage maps were constructed using 435 microsatellite markers genotyped in a full-sib family of the fish species. The female linkage map contained 140 microsatellite markers, in which 23 linkage groups had a total genetic length of 1334.1 c M and average inter-marker space of 13.3 c M. The male linkage map contained 156 microsatellite markers, in which 25 linkage groups had a total genetic length of 1359.6 c M and average inter-marker distance of 12.4 c M. The genome coverage of the female and male linkage maps was 68.6% and 69.3%, respectively. The female-to-male ratio of the recombination rate was approximately 1.07:1 in adjacent microsatellite markers. This paper presents the first genetic linkage map of microsatellites in black rockfish. The collection of polymorphic markers and sex-specific linkage maps of black rockfish could be useful for further investigations on parental assignment, population genetics, quantitative trait loci mapping, and marker-assisted selection in related breeding programs.

Feasibility of Partial Replacement of Fishmeal with Proteins from Different Sources in Diets of Korean Rockfish(Sebastes schlegeli)

An 8-week feeding experiment was conducted in an indoor recirculation seawater system to investigate the effects of partial replacement of dietary fishmeal with proteins from five sources on the growth performance and feed utilization of Sebastes schlegeli. Six isonitrogenous and isoenergetic diets were formulated using fishmeal(FM, the control) as sole protein source, or proteins from five sources including poultry by-product meal(PBM), meat and bone meal(MBM), soybean meal(SBM), cottonseed meal(CSM) and canola meal(CNM). Fifteen percent of the crude protein provided by fish meal was replaced, respectively. The results showed that the differences in specific growth rate(SGR) and survival rate(SR) among fish fed PBM, MBM, SBM, CSM and whole FM diets were not significant. However, SGR and SR of fish fed CNM diet was significantly lower than that of other treatments. Feeding rate, feed conversion, nutrient retention showed similar patterns to that of growth. Fish fed CSM and CNM showed significantly lower apparent digestibility coefficient(ADC) of dry matter and gross energy than those fed others while fish fed CNM showed lower ADC of crude protein than those fed others(P<0.05). These results suggested that it was feasible to substitute 15% dietary protein provided by fishmeal with PBM, MBM, SBM and CSM, respectively, but not with CNM as the replacement with CNM reduced fish growth and feed utilization.

The influence of main ecological and environmental factors on the energy budget of Schlegel＇ s black rockfish Sebastes schlegeli

In the Maidao Ecological Laboratory of the Huanghai Sea Fisheries Research Institute in 1998 ~ 2000, the continuous flow-through method was adopted to determine the energy budget components and models of Schlegel＇ s black rockfish under different ecological and environmental factors, such as temperature, food species, body mass and feeding level, etc. The body mass of test fish ranged from 21.2 to 224.5 g. The specific growth rate tended to invert ＂U＂ change with temperature increase. The ecological conversion efficiency showed the same trend as temperature and feeding level rise. Only the specific growth rate tended to decelerate the increment with the rise of feeding level. From the above quantitative relationships, the maximum and optimal growth temper- atures could be obtained as 17.5 and 16.2 ℃, and the maintenance feeding rate and optimum feeding rate as 0.8% and 4. 1% of the body mass. The specific growth rate and ecological conversion coefficient showed a decelerating decrease trend with body mass increment. The small-sized fish food was more advantageous to the feeding level increase in comparison with the small-sized shrimp food, which led to the increase of growth, metabolism and excretion, but had no significant influence on the ecological conversion coefficient. All of temperature, feeding level, body mass and food species can obviously change the energy budget models. The assigning rate of growth energy tended to change along the invert ＂U＂ curve with the increase of temperature and feeding level, and decrease with the body mass increase, but the assigning rates of metabolism energy and excretion energy were quite reverse. These relationships could be quantitatively described as quadratic or power function curve. If the above relationships proved true to other marine fish, the establishment of energy budget model of the same fish under different conditions can be largely simplified.

Preliminary Study on the Water Drag of Schlegel's Black Rockfish Sebastes schlegeli

Drag forces acting on Schlegel’s black rockfish Sebastes schlegeli were studied. A new drag force transducer was designed and used to measure the water drag on Schlegel’s black rockfish in a vertical recirculating flume tank. Fourteen individuals were investigated, yielding two mean drag coefficients referred to the cross sectional area and volume 2/3 respectively at water velocities ranging from 0.3 to 1.0 m s -1 . The drag coefficients can be used for estimating the drag forces acting on Sebastes schlegeli in water.

许氏平鲉提早繁育技术

根据大连地区网箱养殖生产需要,开展许氏平鲉(Sebastes schlegeli)提早繁育生产试验研究。通过控温、控光、营养强化等措施,突破了许氏平鲉亲鱼提早成熟,并确定了合理的温度、光照、盐度、培养密度、饵料系列等人工育苗条件。较常规季节提前2个月培育出73万尾5 cm的商品鱼苗。

Detection of lymphocystis disease virus in Japanese flounder Paralichthys olivaceus and other marine teleosts from northern China

We isolated a strain of lymphocystis disease virus （LCDV） from Japanese flounder （Paralichthys olivaceus） cultured in northern China. Based on published sequences of major capsid protein （MCP） gene of LCDV-cn （GenBank： AF126405）, we designed two primer sets P1/P2 and P3/P4. We then used one-step or nested PCR and in-situ hybridization （ISFI） to detect LCDV and identify the target tissues or cells in infected Japanese flounder. The PCR products were positive in purified viral supematant, skin nodules, gut, gill, kidney, spleen, stomach, heart, and liver of Japanese flounder. We compared the DNA sequence with 14 MCP nucleotide sequences from GenBank, including Megalocytivirus （OFIV and RSIV）, lridovirus （CzlV and W/V）, Ranavirus （TFV and FV3）, and Lymphocystivirus （8 LCDV）. Based on the alignment, we confirmed the PCR product was from Lymphocystivirus （GenBank accession number DQ279090 （LCDV-HD））. Using ISH, we noted the presence of LCDV in the skin nodules, gut, gill, spleen, stomach, and heart of spontaneously infected Japanese flounders. We successfully amplified LCDV fragments from Schlegel＇s black rockfish （Sebastes schlegeli Higendorf）, redwing sea robin （Lepidotrigla microptera Gtinther） and turbot （Scophthalmus maximus） using the one-step and nested PCR, suggesting the target genes can be widely detected in fish using this method.