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Zhang S.,Marine Fisheries Research Institute of Zhejiang Province | Zhang S.,Zhejiang Ocean University | Guo Y.,Marine Fisheries Research Institute of Zhejiang Province | Yan Z.,Marine Fisheries Research Institute of Zhejiang Province | And 2 more authors.
Analytical and Bioanalytical Chemistry | Year: 2015

Reliably detecting nitrofurazone (NFZ) residues in farmed crab and shrimp was previously hindered by lack of appropriately specific analytical methodology. Parent NFZ rapidly breaks down in meat, and the commonly used side-chain metabolite, semicarbazide (SEM), is non-specific as it occurs naturally in crustacean shell often leading to 'false positive' detections in meat. Using 5-nitro-2-furaldehyde (NF) as marker metabolite, following pre-column derivatization with 2,4-dinitrophenylhydrazine (DNPH), ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis in negative electrospray ionization mode enabled confirmation of NFZ residues in deliberately treated whole crab, crab meat and shrimp meat, with a limit of detection (LOD) and limit of quantification (LOQ) below 1 ng g-1. Meanwhile, the derivatives of DNPH-NF were synthesized for the first time, purified by preparative liquid chromatography and structure characterized with nuclear magnetic resonance spectroscopy (1H-NMR). The purity of derivative was checked by ultra-performance liquid chromatography-tunable ultraviolet (UPLC-TUV), and the contents were beyond 99.9 %. For comparison purposes, crustacean samples were analysed using both NF and SEM marker metabolites. NFZ treatment was revealed by both NF and SEM marker metabolites, but untreated crab also showed measurable levels of SEM which could potentially be misinterpreted as evidence of illegal NFZ use. [Figure not available: see fulltext.] © 2015 Springer-Verlag Berlin Heidelberg.


PubMed | Zhejiang Ocean University and Marine Fisheries Research Institute of Zhejiang Province
Type: Journal Article | Journal: Journal of experimental zoology. Part A, Ecological genetics and physiology | Year: 2016

Sex steroid hormones are widely detected in molluscs and play important roles in sex determination, gonadal tissue maturation, and gametogenesis. Nevertheless, the signaling pathways of sex steroids in cephalopod have not yet been clearly elucidated. In the present study, a full-length sequence encoding the estrogen receptor (ER) was isolated from common Chinese cuttlefish, Sepiella japonica. The sjER cDNA clone was found to contain 1,788 nucleotides including a 1,470 bp open reading frame encoding 489 amino acid (aa) residues. The deduced ER protein consisted of six nuclear receptor characteristic domains. Based on a phylogenetic analysis, the ER DNA-binding domain and ligand-binding domain are highly conserved compared to other mollusc ERs. Highest aa identities were found for sjER with common octopus (Octopus vulgaris) ER (89%) and pacific oyster (Crassostrea gigas) ER (61%). Tissue expression analysis confirmed that sjER was widely distributed among tissues and predominantly expressed in the brain, liver, gonad (testis and ovary), and other accessory sexual gland (nidamental gland). The ER expression was temporally upregulated in the brain, liver, and ovary during the early sexual maturation period in S. japonica, which is coincident with the fluctuation of ovary estradiol content. These suggest that sjER may be involved in regulating the reproductive cycle of S. japonica. A fusion protein transient transfections assay showed that sjER was mainly located in the nucleus, suggesting a possible orthodox working mechanism of S. japonica ER in the nucleus through a ligand-dependent activation of specific gene transcription.


PubMed | National Taiwan Ocean University, Chung Yuan Christian University and Marine Fisheries Research Institute of Zhejiang Province
Type: | Journal: Mitochondrial DNA | Year: 2015

In this study, the complete mitogenome sequence of Redbelly yellowtail fusileer, Caesio cuning (Teleostei: Caesionidae), has been sequenced by the next-generation sequencing method. The assembled mitogenome, consisting of 16 508bp, had the typical vertebrate mitochondrial gene arrangement, including 13 protein-coding genes, 22 transfer RNAs, 2 ribosomal RNAs genes and a non-coding control region of D-loop. D-loop contains 826bp length that is located between tRNA-Pro and tRNA-Phe. The overall base composition of C. cuning is 28.1% for A, 31.1% for C, 16.3% for G and 24.4% for T. The complete mitogenome may provide essential and important DNA molecular data for further phylogenetic and evolutionary analysis for Caesionidae.


Xiao J.-X.,Zhejiang University | Zhou F.,Zhejiang Fisheries Technical Extension Station | Yin N.,Zhejiang University | Zhou J.,Zhejiang University | And 4 more authors.
Aquaculture Research | Year: 2013

An 8-week feeding trial was conducted to evaluate compensatory growth of juvenile black sea bream, Acanthopagrus schlegelii (initial weight 9.56 ± 0.12 g) in fifteen 300 L indoors flow-through circular fibreglass tanks. Feeding regimes was designed as follows: the control group (fed continuously), and S1, S2, S3 and S4 groups experienced 1, 2, 3 and 4 days of feed deprivation and then refeeding for the remaining days per week respectively. Changes in body weight, specific growth rate (SGR), feed intake (FI) and feed efficiency ratio (FER) were examined biweekly during the trial. At the end of the cyclical feeding periods, survival was not significantly affected by feeding strategy (P > 0.05). Final body weight of fish in S1 group was significantly higher than the control group (P < 0.05) after an 8 weeks trial, and fish in the S2 group reached the same body weight of the control fish (P > 0.05), however, the growth data in the S3 and S4 groups could not catch-up with the control treatment (P < 0.05). In the first 4 weeks, SGR values of fish with feed deprivation more than 2 days per week were inferior to those of control group (P < 0.05); however, no significant differences of SGR were observed among the groups for the last 4 weeks (P > 0.05). FI increased significantly with the starvation days increasing during the whole feeding trial (P < 0.05). At the periods 2, 4 and 6 weeks, FER values increased significantly with increasing feed-deprivation days up to S2 group and then levelled off (P < 0.05); however, FER in S2 group was only higher than that in S0 and S1, but no significant differences were found among the other treatments at the end of week 8. Significant differences were found in apparent digestibility coefficients (ADCs) of dry matter and crude protein among the treatments, while ADCs of crude lipid were unaffected. Protein and lipid contents in whole body and dorsal muscle showed declining tendency with increasing food deprivation days, while moisture contents tended to increase (P < 0.05). Serum parameters were markedly affected by feeding regimes except for total protein concentration and thyroxine level. The present results indicated that starvation for 1 and 2 days per week of juvenile black sea bream could achieve over-compensation and complete compensation respectively. However, in case of longer term feed restriction regime, fish failed to obtain good growth performance. © 2013 John Wiley & Sons Ltd.


Chai X.J.,Marine Fisheries Research Institute of Zhejiang Province | Ji W.X.,Zhejiang University | Han H.,Zhejiang University | Dai Y.X.,Zhejiang University | Wang Y.,Zhejiang University
Aquaculture Nutrition | Year: 2013

A 50-day feeding trial was conducted to examine the effects of dietary protein and lipid levels on growth, feed utilization, body composition and swimming performance of giant croaker, Nibea japonica. Fish (initial body weight 44.6 g ind-1) were fed ten test diets which were formulated at 5 crude protein levels (360, 400, 440, 480 and 520 g kg-1) and 2 crude lipid levels (90 and 150 g kg-1). In addition, a raw fish diet (fillet of small yellow croaker) served as the reference. The weight gain (WG) increased, whereas the feed intake (FI) and feed conversion ratio (FCR) decreased, with increasing dietary protein level from 360 to 520 g kg-1. At the same dietary protein level, no significant difference was found in the WG between fish fed the diets containing 90 or 150 g kg-1 crude lipid. Fish fed the diet containing 480 g kg-1 crude protein and 90 g kg-1 crude lipid exhibited higher WG, nitrogen retention efficiency (NRE) and energy retention efficiency (ERE) but lower nitrogen wastes output (TNW). At the end of the feeding trial, the hepatosomatic index (HSI) and viscerosomatic index (VSI) decreased, whereas the body protein content increased, with increase in dietary protein level. The body lipid content was higher in fish fed at the 150 g kg-1 lipid level than in fish fed at the 90 g kg-1 lipid level. No significant difference was found in the maximum sustained swimming speed (MSS) between fish fed at different dietary protein and lipid levels. The WG, NRE, ERE and condition factor (CF) were higher, whereas the FI, FCR, HSI, VSI and TNW were lower, in fish fed the raw fish diet than in fish fed the diet containing 480 g kg-1 crude protein and 90 g kg-1 crude lipid. No significant difference was detected in the MSS between fish fed the raw fish diet and diet containing 480 g kg-1 crude protein and 90 g kg-1 crude lipid. The results of this study suggest that the suitable dietary crude protein and crude lipid levels are 480 g kg-1 and 90 g kg-1 for giant croaker reared in net pens. © 2013 John Wiley & Sons Ltd.


Zhou F.,Zhejiang University | Xiong W.,Zhejiang University | Xiao J.-X.,Zhejiang University | Shao Q.-J.,Zhejiang University | And 3 more authors.
Aquaculture Research | Year: 2010

An 8-week feeding trial was conducted to determine the dietary arginine requirement of juvenile black sea bream Sparus macrocephalus in 18 350 L indoors flow-through circular fibreglass tanks. Six isonitrogenous and isoenergetic diets were formulated to contain graded levels of l-arginine (1.85%, 2.23%, 2.51%, 2.86%, 3.20% and 3.46% dry diet) from dietary ingredients and crystalline arginine. Each diet was randomly assigned to triplicate groups of 25 juvenile fish (10.51±0.15 g) twice daily (08:00 and 16:00 hours) to apparent satiation. Results showed that the specific growth rate (SGR) increased with increasing dietary arginine levels up to 2.51% and remained nearly the same thereafter. Feed efficiency ratio, protein efficiency ratio (PER) and protein productive value all showed an increasing tendency and then levelled off. Apparent digestibility coefficients of dry matter, crude protein and gross energy significantly improved up to 2.86% arginine diet and decreased at different extents thereafter. Fish fed 1.85% arginine diet had significantly lower protein content in the whole body and dorsal muscle than those fed diets supplemented with or >2.86% of arginine. Lipid content decreased and lower value occurred at 3.46% of dietary arginine. The dietary essential amino acid composition in the whole body of the black sea bream was significantly influenced by dietary arginine. Arginine retention increased with an increasing dietary arginine level from 1.85% to 3.20%, then declined slightly at 3.46% arginine diet. Serum biochemical parameters were significantly affected by the dietary arginine level except for the cholesterol content. Broken-line regression based on SGR and second-order polynomial regression based on PER indicated that the optimum dietary arginine requirements for juvenile black sea bream were 2.79% and 3.09% diet, corresponding to 7.74% and 8.13% of the dietary protein respectively. © 2010 The Authors. Aquaculture Research © 2010 Blackwell Publishing Ltd.


Zhang J.,Zhejiang University | Zhou F.,Zhejiang University | Wang L.-L.,Zhejiang University | Shao Q.,Zhejiang University | And 2 more authors.
Journal of the World Aquaculture Society | Year: 2010

An 8-wk experiment was conducted to determine the optimal protein requirement of juvenile black sea bream, Sparus macrocephalus, (initial weight 13.13 ± 0.21 g, mean ± SD) in 18 300-L indoor flow-through circular fiberglass tanks provided with sand-filtered aerated seawater. Six isoenergetic diets were formulated to contain varying protein levels ranging from 31.95 to 48.53% at about 3% protein increments by substituting corn oil and α-starch for fish meal. Each diet was assigned to triplicate groups of 20 fish in a completely randomized design. Twenty uniform-sized fish were stocked in a 300-L indoor flow-through circular fiberglass tank. After the feeding trial, weight gain and specific growth rate (SGR) increased with increasing levels of dietary protein up to 41.8% (P < 0.05) and both showed a declining tendency thereafter. Survival could not be related to dietary treatments. Both condition factor and protein efficiency ratio declined while dietary protein levels increased (P < 0.05). The highest and the lowest values of hepatosomatic index were found in fish fed 31.9 and 41.8% protein diets (P < 0.05), respectively, and showed no significant difference among other treatments (P > 0.05). Feed efficiency ratio (FER) improved as dietary protein level increased, with the maximum FER in the 41.8% protein diet, although this was not significantly different from the 45.2% protein diets. Apparent digestibility coefficients (ADCs) of main ingredients of diets tended to improve as the protein level increased, although they decreased to a different extent at higher levels of dietary protein. The highest ADC values of organic matter and protein were both found for fish fed 41.8% protein diet, while lipid and energy were at 38.5 and 45.2% protein diet, respectively. The whole-body protein content was positively correlated with dietary protein levels, while lipid content was negatively correlated. The muscle crude protein and crude lipid contents were significantly affected (P < 0.05) by dietary protein level, while moisture and ash showed no significant differences (P > 0.05). Dietary protein levels had significant influences on plasma parameters (P < 0.05). The concentrations of total cholesterol, triacylglyceride, alanine transaminase, and aspartate transaminase significantly reduced with dietary protein level except plasma total protein contents which presented reverse trend. Analysis of dose (protein level)-response (SGR) with second-order polynomial regression suggested that the optimal dietary protein requirement was 41.4%. © Copyright by the World Aquaculture Society 2010.


Zhou F.,Zhejiang University | Shao J.,Zhejiang University | Xu R.,Zhejiang University | Ma J.,Zhejiang University | Xu Z.,Marine Fisheries Research Institute of Zhejiang Province
Aquaculture Nutrition | Year: 2010

An 8-week feeding experiment was conducted to determine the quantitative l-lysine requirement of juvenile black sea bream Sparus macrocephalus (initial mean weight: 9.13 ± 0.09 g, SD) in eighteen 300-L indoors flow-through circular fibreglass tanks provided with sand-filtered aerated seawater. The experimental diets contained six levels of l-lysine ranging from 20.8 to 40.5 g kg-1 dry diet at about 4 g kg-1 increments. All the experiment diets were formulated to be isoenergetic and isonitrogenous. Each diet was assigned to triplicate groups of 20 fish in a completely randomized design. Weight gain and specific growth rate (SGR) increased with increasing levels of dietary lysine up to 32.5 g kg-1 (P < 0.05) and both showed a declining tendency thereafter. Feed efficiency ratio and protein efficiency ratio was poorer for fish fed the lower lysine level diets (P < 0.05) and showed no significant differences among other treatments (P > 0.05). All groups showed high survival (above 90%) and no significant differences were observed. The whole body crude protein and crude lipid contents were significantly affected (P < 0.05) by dietary lysine level, while moisture and ash showed no significant differences. The composition of muscle and liver also presented similar change tendency. Total essential amino acid and lysine contents in muscle both obtained the highest value when fish fed 32.5 g kg-1 lysine diet (P < 0.05). Serum protein, cholesterol and free lysine concentration were affected by different dietary treatments (P < 0.05), triacylglyceride and glucose contents were more variable and could not be related to dietary lysine levels. Dietary lysine level significantly affected condition factor and intraperitoneal fat ratio of juvenile black sea bream (P < 0.05) except for hepatosomatic index. There were no significant differences in white blood cell count and red blood cell count (P > 0.05), however, haemoglobin level was significantly influenced by different diets (P < 0.05). Analysis of dose (lysine level)-response (SGR) with second order polynomial regression suggested the dietary lysine requirement of juvenile black sea bream to be 33.2 g kg-1 dry diet or 86.4 g lysine kg-1 protein. © 2009 Blackwell Publishing Ltd.


PubMed | Marine Fisheries Research Institute of Zhejiang Province
Type: | Journal: Analytical and bioanalytical chemistry | Year: 2017

An ultraperformance liquid chromatography-quadrupole time-of-flight high-resolution mass spectrometry method was developed and validated for the determination of nitrofurazone metabolites. Precolumn derivatization with 2,4-dinitrophenylhydrazine and p-dimethylaminobenzaldehyde as an internal standard was used successfully to determine the biomarker 5-nitro-2-furaldehyde. In negative electrospray ionization mode, the precise molecular weights of the derivatives were 320.0372 for the biomarker and 328.1060 for the internal standard (relative error 1.08ppm). The matrix effect was evaluated and the analytical characteristics of the method and derivatization reaction conditions were validated. For comparison purposes, spiked samples were tested by both internal and external standard methods. The results show high precision can be obtained with p-dimethylaminobenzaldehyde as an internal standard for the identification and quantification of nitrofurazone metabolites in complex biological samples. Graphical Abstract A simplified preparation strategy for biological samples.


PubMed | Marine Fisheries Research Institute of Zhejiang Province
Type: Evaluation Studies | Journal: Analytical and bioanalytical chemistry | Year: 2015

Reliably detecting nitrofurazone (NFZ) residues in farmed crab and shrimp was previously hindered by lack of appropriately specific analytical methodology. Parent NFZ rapidly breaks down in meat, and the commonly used side-chain metabolite, semicarbazide (SEM), is non-specific as it occurs naturally in crustacean shell often leading to false positive detections in meat. Using 5-nitro-2-furaldehyde (NF) as marker metabolite, following pre-column derivatization with 2,4-dinitrophenylhydrazine (DNPH), ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis in negative electrospray ionization mode enabled confirmation of NFZ residues in deliberately treated whole crab, crab meat and shrimp meat, with a limit of detection (LOD) and limit of quantification (LOQ) below 1 ng g(-1). Meanwhile, the derivatives of DNPH-NF were synthesized for the first time, purified by preparative liquid chromatography and structure characterized with nuclear magnetic resonance spectroscopy ((1)H-NMR). The purity of derivative was checked by ultra-performance liquid chromatography-tunable ultraviolet (UPLC-TUV), and the contents were beyond 99.9%. For comparison purposes, crustacean samples were analysed using both NF and SEM marker metabolites. NFZ treatment was revealed by both NF and SEM marker metabolites, but untreated crab also showed measurable levels of SEM which could potentially be misinterpreted as evidence of illegal NFZ use.

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