Artificial induction of mito-gynogenetic diploids in large yellow croaker (Pseudosciaena crocea) by hydrostatic pressure

The present study investigated conditions for inducing mito-gynogenetic(endomitosis) diploids by hydrostatic pressure in the large yellow croaker Pseudosciaena crocea.In haploid control groups,the development of eggs was activated with ultraviolet radiated semen.All fry presented typical haploid syndrome in the haploid control groups,and were verified as haploids using cytometry.After hydrostatic pressure treatment,morphologically normal fry reappeared at different frequencies according to the intensity and...     

Artificial induction of mito-gynogenetic diploids in large yellow croaker (Pseudosciaena crocea) by hydrostatic pressure

The present study investigated conditions for inducing mito-gynogenetic (endomitosis) diploids by hydrostatic pressure in the large yellow croaker Pseudosciaena crocea. In haploid control groups, the development of eggs was activated with ultraviolet radiated semen. All fry presented typical haploid syndrome in the haploid control groups, and were verified as haploids using cytometry. After hydrostatic pressure treatment, morphologically normal fry reappeared at different frequencies according to the intensity and time of pressure shock. Fry with normal appearance in the pressure treated groups were verified as gynogenetic double haploids (GDHs), containing only one allele from the female parent at all four diagnostic microsatellite loci. For a fixed duration of 3 min, the optimal intensity of blocking the first mitosis was determined to be 40 Mpa, which was similar to that of blocking the second meiosis. There was a “window” of starting time, from 36.1 min to 38.1 min post-insemination at 25.0±1.0°C, within which the production of GDHs was not significantly different. Maximum production of morphologically normal fries, 9.36%±2.97% of developed eggs, was found when the eggs were shocked with hydrostatic pressure at 40 Mpa for 3 min, starting from 38.1 min post insemination at 25.0±1.0°C.     

Effect of dietary olaquindox on the growth of large yellow croaker (Pseudosciaena crocea R.) and the distribution of its residues in fish tissues

Olaquindox(OLA), one of quinoxaline-N, N-dioxides, has been put under ban. However it was used as a medicinal feed additive early; it promotes the growth of livestock and prevents them from dysentery and bacterial enteritis. In this study, we evaluated the effect of dietary OLA on the growth of large yellow croaker(Pseudosciaena crocea R.) and the histological distribution of OLA and its metabolite 3-methyl-quinoxaline-2-carboxylic acid(MQCA) in fish tissues. Four diets containing 0(control), 42.5, 89.5 and 277.2 mg kg-1 OLA, respectively, were formulated and tested, 3 cages(1.0 m × 1.0 m × 1.5 m) each diet and 100 juveniles(9.75 ± 0.35 g) each cage. The fish were fed to satiation twice a day at 05:00 am and 17:00 pm for 8 weeks. The survival rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet containing 0 and 277.2 mg kg-1 OLA(P 0.05), while the weight gain rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet without OLA(control)(P0.05), but similar to that of fish fed the diet with 277.2 mg kg-1 OLA. Fish fed the diet with 277.2 mg kg-1 OLA had the highest content of OLA and MQCA in liver(3.44 and 0.39 mg kg-1, respectively), skin(0.46 and 0.09 mg kg-1, respectively) and muscle(0.24 and 0.06 mg kg-1, respectively). In average, fish fed the diet containing OLA had the highest content of OLA and MQCA in liver which was followed by skin and muscle(P 0.05), whereas OLA and MQCA were not detectable in control. Our findings demonstrated that OLA and MQCA accumulated in large yellow croaker when it was fed with the diet containing OLA, thus imposing a potential safety risk to human health.     

Establishment of a spleen cell line from large yellow croaker Pseudosciaena crocea and its primitive application in foreign gene transfection

A large yellow croaker,Pseudosciaena crocea,spleen(LYCS)cell line was established and the feasibility of using it for foreign gene transfection was evaluaed in this study.Primary culture of LYCS cells was initiated from spleen tissue pieces,which were cultured at 25℃ in Dulbecco’s modiced Eagle medium/F12 medium(DMEM/F12,1:1)(pH7.2),supplemented with 20% fetal bovine serum,carboxymethyl chitosan,chondroitin sulfate,basic fibroblast growth factor(bFGF)and insulin-like growth factor-I(IGF-I).The cultured LYCS cells,in fibroblast shape,proliferated to 100% confluency 20 days later.Chromosome analyses indicated that the LYCS cells exhibited chromosomal aneuploidy with a modal chromosome number of 48 which displayed the normal diploid karyotype of P.crocea(6m+6sm+36t,NF=60).A LYCS cell line,with a population doubling time of 48.7 h at passage 60,has been established and subcultured to passage 70.Transgenic feasibility test demonstrated that positive green fluorescence protein(GFP)expression was observed in LYCS cells after pcDNA3.1-GFP plasmid transfection.In conclusion,a continuous foreign gene trans-fection feasible LYCS cell line has been established successfully.The cell line might serve as a valuable tool for studies of transgenic breeding and has potential applications for different kinds of cytotechnological studies.     

Comparative study on the organoleptic quality of wild and farmed large yellow croaker Larimichthys crocea

Journal of Oceanology and Limnology - To fill the blank in systematic study on the organoleptic quality of large yellow croaker Larimichthys crocea, we analyzed the organoleptic quality of the fish...     

Estimated reproductive success of brooders and heritability of growth traits for large yellow croaker (Larimichthys crocea) using microsatellites

We estimated the reproductive success of adult brood fish and the heritability of growth-related traits for large yellow croaker (Larimichthys crocea). We created two independent full-factorial cross groups (1 and 2) by crossing 4 males×4 females and 4 males×3 females, respectively. We measured the body weight (BW), body length (BL), and body height (BH) of 281 individuals from group 1 and 318 individuals from group 2 at 20 months post hatch (harvest age). We also collected a tissue sample from each individual. The parents and offspring were genotyped using six polymorphic microsatellites. Of the 599 offspring, 99.2% were assigned to a single pair of parents. In both groups, some parent pairs produced a large number of offsprings while other pairs did not produce any offspring. The genetic diversity and putative Ne were lower in the offsprings than in the parents in both groups. The heritability estimates at 20-month age were 0.13±0.10 for BW, 0.19±0.13 for BL, and 0.09±0.06 for BH. The genetic and phenotype correlation between BW, BL, and BH was close to 1. The results provide basic information for selective breeding and further genetic characterization of large yellow croaker.     

Replacement of Dietary Fish Oil with Vegetable Oils Improves the Growth and Flesh Quality of Large Yellow Croaker （Larmichthys crocea）

We investigated the effect of the replacement of dietary fish oil with vegetable oils on the growth and flesh quality of large yellow croaker(Larmichthys crocea). The basal diet(FO) was formulated to contain 66.5% fish meal and 6.4% menhaden fish oil; whereas the other 3 experimental diets were formulated by replacing the fish oil with 50% soybean oil(SO50), 100% soybean oil(SO100) and 100% palm oil(PO100), respectively. The 4 diets were randomly assigned to 4 floating sea cages(3.0 m × 3.0 m × 3.0 m), and each was stocked with 250 fish individuals with an initial average weight of 245.29 g ± 7.45 g. The fish were fed to apparent satiation twice a day at 5:00 and 17:00, respectively, for 12 weeks. Experimental analysis showed that the specific growth rate of fish fed SO50 or PO100 were significantly higher than that of fish fed FO or SO100(P0.05), and crude lipid contents of ventral muscle and viscera were significantly lower in fish fed FO than in those fed the other 3 diets(P0.05). No significant differences in condition factor, viscerosomatic index, hepatosomatic index, gutted yield and colorimetric values of fish among the dietary treatments were observed(P0.05). Compared to FO diet, SO50, SO100 and PO100 diets led to substantial decreases in the liquid loss and water loss from fresh fillets(1 d, 4℃)(P0.05). Similarly, thiobarbituric acid reactive substance(TBARS) values of fillets under different storage conditions(1 d, 4℃; 7 d, 4℃; 4 weeks,-20℃; 8 weeks,-20℃) decreased significantly after partial or complete replacement of fish oil with vegetable oils. These findings indicated that the growth performance and selected flesh quality properties(liquid holding capacity and TBARS value) of large yellow croaker were substantially improved by replacing dietary fish oil with vegetable oils.     

Thermal tolerance evaluation and related microsatellite marker screening and identification in the large yellow croaker Larimichthys crocea

Thermal tolerance to high temperature was evaluated in the large yellow croaker Larimichthys crocea. The survival thermal maximum for L. crocea was 33.0°C, the 50% critical thermal maximum(50% CTMax) was 35.5°C, and the critical thermal maximum(CTMax) was 36.0°C. Three microsatellite markers(LYC0148, LYC0200 and LYC0435), associated with thermal tolerance were screened and identified using a Bulked Segregation Analysis(BSA) method. These markers have six amplified fragments in which four are related to thermal tolerance. These fragments were cloned and sequenced, and the results showed the core motif were all "AC" repeats. For LYC0148 and LYC0200, the lengths of fragments are 181 bp and 197 bp, respectively. For LYC0435, which has two fragments, the fragment lengths are 112 bp and 100 bp. The results provide useful molecular markers for thermal-tolerance breeding of large yellow croaker in the near future.     

Dietary leucine requirement for juvenile large yellow croaker <Emphasis Type="Italic">Pseudosciaena crocea</Emphasis> (Richardson, 1846)

Dietary leucine requirement for juvenile large yellow croaker, Pseudosciaena crocea Richardson 1846 (initial body weight 6.0 g ± 0.1 g) was determined using dose-response method. Six isonitogenous (crude protein 43%) and isoenergetic (19 kJ g⁻¹) practical diets containing six levels of leucine (Diets 1–6) ranging from 1.23% to 4.80% (dry matter) were made at about 0.7% increment of leucine. Equal amino acid nitrogen was maintained by replacing leucine with glutamic acid. Triplicate groups of 60 individuals were fed to apparent satiation by hand twice daily (05:00 and 17:30). The water temperature was 26–32°C, salinity 26–30 and dissolved oxygen approximately 7 mg L⁻¹ during the experimental period. Final weight (FW) of large yellow croaker initially increased with increasing level of dietary leucine but then decreased at further higher level of leucine. The highest FW was obtained in fish fed diet with 3.30% Leucine (Diet 4). FW of fish fed the diet with 4.80% Leucine (Diet 6) was significantly lower than those fed Diet 4. However, no significant differences were observed between the other dietary treatments. Feed efficiency (FE) and whole body composition were independent of dietary leucine contents (P > 0.05). The results indicated that leucine was essential for growth of juvenile large yellow croaker. On the basis of FW, the optimum dietary leucine requirement for juvenile large yellow croaker was estimated to be 2.92% of dry matter (6.79% of dietary protein).     

Tyrosinase, a new innate humoral immune parameter in large yellow croaker （Pseudosciaena crocea R）

We evaluated the immune response to infection with a pathogen in large yellow croaker(Pseudosciaena crocea Richardson).The fish were given an intraperitoneal(i.p.) injection of Vibrio parahaemolyticus or sterile sea water(control).We collected blood sera from the fish 0.17,1,2,4,8,12,or 16 d after injection(dpi).We measured tyrosinase activity and the concentrations of lysozyme,NOS,and antibodies.Serum tyrosinase activity was significantly higher at 0.17 and 4 dpi than in the control group,and peaked at 8 d...     

Dietary Leucine Requirement for Juvenile Large Yellow Croaker Pseudosciaena crocea (Richardson,1846)

Dietary leucine requirement for juvenile large yellow croaker, Pseudosciaena crocea Richardson 1846 (initial body weight 6.0 g±0.1 g) was determined using dose-response method.Six isonitogenous (crude protein 43%) and isoenergetic (19 kJ g-1) practical diets containing six levels of leucine (Diets 1-6) ranging from 1.23% to 4.80% (dry matter) were made at about 0.7% increment of leucine.Equal amino acid nitrogen was maintained by replacing leucine with glutamic acid.Triplicate groups of 60 individuals were fed to apparent satiation by hand twice daily (05:00 and 17:30).The water temperature was 26-32℃, salinity 26-30 and dissolved oxygen approximately 7 mg L-1 during the experimental period.Final weight (FW) of large yellow croaker initially increased with increasing level of dietary leucine but then decreased at further higher level of leucine.The highest FW was obtained in fish fed diet with 3.30% Leucine (Diet 4).FW of fish fed the diet with 4.80% Leucine (Diet 6) was significantly lower than those fed Diet 4.However, no significant differences were observed between the other dietary treatments.Feed efficiency (FE) and whole body composition were independent of dietary leucine contents (P0.05).The results indicated that leucine was essential for growth of juvenile large yellow croaker.On the basis of FW, the optimum dietary leucine requirement for juvenile large yellow croaker was estimated to be 2.92% of dry matter (6.79% of dietary protein).     

大黄鱼库道虫病的初步研究

在广东湛江海水网箱养殖的大黄鱼 (Pseudosciaenacrocea)中发现了库道虫病 (Kudoasis) ,检出率高达 31%。其症状表现为 :白色孢囊分布于胸鳍、腹鳍、臀鳍和尾鳍等的鳍膜下 ,剖检时发现腹壁和背部肌肉中布满许多白色呈球形或椭圆形的小孢囊 ,在肠系膜、腹膜以及近口腔的粘膜上也有大量的白色小孢囊分布。镜检发现孢囊中含有典型的库道虫 ,其孢子宽为 ( 8 90± 0 55) μm ,长为 ( 8 35± 0 4 6 ) μm ,极囊宽为 ( 2 39± 0 36 ) μm ,极囊长为 ( 4 4 8± 0 39) μm。孢子在淡水中 2 0min后全部死亡 ,在盐度为 6的水体中 ,12h和 2 4h后存活率分别约为 87.4 %和 70 % ;在盐度为 16、2 6和 36的水体中 ,孢子存活率均为 10 0 % ;在盐度为 4 6的水体中 ,12h和 2 4h后存活率分别约为92 %和 84 %。孢子对低温有较强的耐受力 ,当水温达到 4 4℃时 ,8h、12h和 2 4h后 ,孢子的存活率分别约为 91%、80 .5%和 6 6 %。     

Immune response of Pseudosciaena crocea to the injection of Vibrio alginolyticus

For the investigation of anti-infection immune response of Pseudosciaena crocea, 160 healthy fish samples were categorized into infected and control groups. Each individual fish in the infected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension of Vibrio alginolyticus in density of 2×107 CFU/ml, while each individual in the control group was injected i.p. with 0.2 ml sterile saline solution (0.85%). It was observed that the artificial injection of V. alginolyticus significantly inc...     

Lipid oxidation and fatty acid composition in salt-dried yellow croaker (<Emphasis Type="Italic">Pseudosciaena polyactis</Emphasis>) during processing

Lipid oxidation in salt-dried yellow croaker (Pseudosciaena polyactis) was evaluated during processing with commonly used analytical indices, such as the peroxide value (POV), the thiobarbituric acid reactive substances (TBARS) value, and oxidative-relative lipoxygenase (LOX) activity. Additionally, fatty acids were analyzed using gas chromatography-mass spectrometry. Both POV and TBARS increased significantly (P < 0.05) at the rinsing stage. POV reached its peak value of 3.63 meq O₂ per kg sample at the drying stage, whereas TBARS constantly increased from 0.05 to 0.20 mg MDA per kg sample. Processing of salt-dried yellow croaker had an extremely significant (P < 0.01) effect on LOX activity. Twenty-six fatty acids were identified. Combined eicosapentaenoic acid (EPA; C20:5n3) and docosahexaenoic acid (DHA; C22:6n3) content varied between (19.20 ± 0.37) mg g^?1 and (23.45 ± 1.05) mg g^?1. The polyunsaturated fatty acid/saturated fatty acid (PUFA/SFA) ratio in yellow croaker was 0.73–1.10, and the n-6/n-3 PUFA ratio was approximately 0.13–0.20. The contents of most fatty acids varied significantly (P < 0.05) during the different processing stages, and these differences were caused by lipid oxidation. C18:0, C16:1n7, C19:0, and C22:6n3 showed clear changes in principle component one of a principle components analysis. These fatty acids are potential markers for evaluating lipid oxidation in fish muscle because there was a significant correlation between these markers and TBARS and LOX activity (P < 0.05) with Pearson’s coefficients > 0.931.     

A Fast and Indirect Fluorescent Antibody Assay for the Vibrio in Large Yellow Croaker Pseudosciaena crocea (Richardson)

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Growth and mortality of small yellow croaker (Larimichthys polyactis) inhabiting Haizhou bay of China

Length frequency data of small yellow croaker(Larimichthys polyactis) were acquired from the survey vessel in May,July,September and December,2011 in Haizhou Bay of China.In this study,921 fish individuals were analyzed for the estimation of growth and mortality parameters.Between length and weight,the power coefficient b was 2.7321,2.9703,3.0418 and 2.7252 for the 4 surveying months,respectively.The estimated von Bertalanffy growth function parameters were 230mm(L ∞) and 0.26yr-1(K) as were calculated with ELEFAN method equipped in FiSAT computer package.With length-converted catch curve analysis,the total mortality rate(Z) and its 95% confidence interval were 2.16(1.69-2.64) yr-1,0.59(0.15-1.04) yr-1,1.16(0.80-1.52) yr-1 and 0.96(0.70-1.23) yr-1 for the 4 surveying months,respectively,with the pooled data the value was 1.15(0.81-1.48) yr-1.The natural mortality rate(M) was 0.516 yr-1 as was calculated with Pauly’s equation(the annual average sea water temperature was 11℃).Therefore,fish mortality rate was 0.634 yr-1.The yield-per-recruit analysis indicated that when t c was 1,F max was 0.7 and F 0.1 was 0.55.Currently,the age at first capture is about 1 year and F current was 0.634.Therefore,F current was larger than F 0.1 and less than F max.This indicates that current fish mortality is at a dangerously high level.With Gulland method,the biological reference point for fishery(F opt) was estimated as 0.516 yr-1,lower than current fish mortality.Accordingly,reducing catch in the region was strongly recommended.     

Effect of twine thickness on size-selectivity of driftnet for the yellow croaker Larimichthys polyactis in southwestern Sea of Korea

In order to determine the effect of twine thickness on the size-selectivity of the driftnet used for the yellow croaker, size-selectivity tests were conducted with three different twine thicknesses(monofi lament diameters of 0.279 mm(number's method; No. 3), 0.321 mm(No. 4), and 0.360 mm(No. 5)) of driftnets for the yellow croaker in the seas around Chooja-do, Jeju Islands. The selectivity curve was estimated by using Kitahara's method. In order to determine the physical properties of the twine used in the experimental fi shing nets, we measured the breaking load, elongation, and stiffness under both dry and wet conditions. In terms of physical properties, the thinnest twine(No. 3) had the strongest breaking strength per unit cross-sectional area, along with good elongation and excellent fl exibility. The thickest twine(No. 5) had the lowest fl exibility. In terms of selectivity, the net of No. 3 twine showed the broadest selection range and, thus, a relatively low selectivity compared with the other nets, while the less fl exible net of No. 5 twine showed the narrowest selectivity range and high selectivity. In addition, it was found that a thicker twine resulted in a smaller haul of small fi sh. Therefore, it can be inferred that the thickness of the twine affects the size of the catch and selectivity, and thus the size composition of the catch as well.     

Trophic ecology of small yellow croaker (Larimichthys polyactis Bleeker): stable carbon and nitrogen isotope evidence

The trophic ecology of the small yellow croaker (Larimichthys polyactis) was studied using stable isotope analyses. Samples were collected from July to September 2009 and 34 individuals from eight sites were examined for stable carbon and nitrogen isotopes. Stable carbon isotope ratios (δ13C) ranged from -20.67 to -15.43, while stable nitrogen isotope ratios (δ15N) ranged 9.18-12.23. The relationship between δ13C and δ15N suggested high resource partitioning in the sampling area. Significant differences in stable isotope values among the eight sampling sites may be linked to environmental diversities involving various physical processes (such as ocean current, wind and tide) and different carbon sources. Furthermore, the stable isotope ratios may also explain the ontogenetic variability in diet and feeding, because δ13C and δ15N varied significantly with increasing body size. The findings are consistent with other studies on diet analyses in small yellow croaker. It was also demonstrated that stable isotope analysis could be used to estimate the trophic characters of small yellow croaker in feeding patterns and migrating habits.     

A Fast and Indriect Fluorescent Antibody Assay for the Vibro in Large Yeelow Croaker Pseudosciaena Crocea (Richardson)

A fast and indirect fluorescnet antibody assay for the Vibrio alginolyticus and V.Parahaemolyticus infecting the large yellow croaker has been developed.The specific antisera for the two strains of vibrio were prepared with New Zealand rabbit and the antiserum and cross-reactive efficacy was tested by coagulation in tube.It showed that the goat anti-rabbit IgG had been labeled by fuorescence isothiocyanate(FITC).The results showed that positive reactions were 100% for the large yellow croaker Pseudosciaena crocea with typical symptom of vibrio infection,while the positive reaction to the pathogen in healthy yellow croakers reached 40%,but seemed negative for aquaculter water.The results demonstrated that this fast and indirect fluorescent antibody assay can be used not only to test the vibrio pathogen in diseased yellow croaker but also in infected animals with no symptom.