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The experiment was conducted in a sealed, temperature-controlled room. The initial water temperature of the treatment group was 12°C, lowered by 1°C a day to 8°C using a chiller, and maintained at that temperature for 3 days. The water temperature was then increased by 1°C per day to 12°C using a heater. The control group were the fish that kept at 12°C at the beginning of the experiment; T1 represents the group that sampled on the first day when the water temperature reached 8 from 12°C; T2 represents the group that sampled when the fish had been kept at 8°C for 3 days; T3 represents the group sampled when the water temperature was increased to 12 from 8°C. The antioxidant abilities of the liver, muscle, and intestine of large yellow croaker at different temperatures and times were determined, and the expression of HSP70 and HSP90 in the liver as well as the changes in intestinal microbiota were investigated. The results showed that the activities of SOD, CAT, GSH-Px, and MDA content increased significantly when the water temperature decreased from 12 to 8°C, significantly higher than those in the control group. The gene expression levels of HSP70 and HSP90 in the liver of large yellow croaker significantly increased under low temperature stress and were significantly higher than those in the control group. The diversity and abundance of intestinal microbiota of large yellow croaker were altered when the water temperature was maintained at 8°C, although the change was not obvious. Proteobacteria, Firmicutes, and Bacteroidetes were the dominate phyla in both the 12 and 8°C groups, and the temperature variation did not change the functional pathways of the microbiota. In conclusion, the acute temperature variation affected the antioxidant ability and heat shock protein expression of large yellow croaker but did not affect the diversity or abundance of the intestinal microbiota. These results will provide a theoretical basis for studying the mechanisms of large yellow croaker overwintering and adaptation to low temperatures.

Highlights • Multi-frequency ultrasound assisted freezing accelerated the freezing rate. • Increase in the number of ultrasonic frequencies enhanced the cavitation effects. • TUF treated samples retained better quality attributes. • Light microscopy observation was adopted to study the microstructures of frozen fish., This research evaluated the effects of multi-frequency ultrasound assisted freezing (UAF) on the freezing rate, structural characteristics, and quality properties of cultured large yellow croaker. The freezing effects with triple ultrasound-assisted freezing (TUF) at 20, 28 and 40 kHz under 175 W was more obvious than that of single ultrasound-assisted freezing (SUF) at 20 kHz and dual ultrasound-assisted freezing (DUF) at 20 and 28 kHz. The results showed that UAF significantly increased the freezing rate and better preserved the quality of frozen large yellow croaker samples. Specifically, the quality parameters of the TUF-treated samples were closer to those of the fresh samples, with greater texture characteristics, a larger water holding capacity (lower thawing loss and cooking loss), lower K values and lower thiobarbituric acid reactive substances values. Light microscopy observation images revealed that the ice crystals formed by TUF were fine and evenly distributed, resulting in less damage to the frozen large yellow croaker samples. Therefore, multi-frequency UAF could improve the quality properties of the large yellow croaker samples.

High levels of soybean oil (SO) in fish diets enriched with linoleic acid (LA, 18:2n-6) could induce strong inflammation. However, the molecular mechanism underlying LA-induced inflammation in the liver of large yellow croaker (Larimichthys crocea) has not been elucidated. Based on previous research, autophagy has been considered a new pathway to relieve inflammation. Therefore, the present study was performed to investigate the role of autophagy in regulating LA-induced inflammation in the liver of large yellow croaker in vivo and in vitro. The results of the present study showed that activation of autophagy in liver or hepatocytes could significantly reduce the gene expression of proinflammatory factors, such as tumor necrosis factor α (TNFα) and interleukin 1β (IL1β). The results of the present study also showed that inhibition of autophagy could upregulate the gene expression of proinflammatory factors and downregulate the gene expression of anti-inflammatory factors in vivo and in vitro. Furthermore, autophagy could alleviate LA-induced inflammatory cytokine gene expression in vivo and in vitro, while inhibition of autophagy obtained the opposite results. In conclusion, our study shows that autophagy could regulate inflammation and alleviate LA-induced inflammation in the liver of large yellow croaker in vivo and in vitro for the first time, which may offer considerable benefits to the aquaculture industry and human health.

Free fatty acid receptor 4 (FFAR4) plays a key role in regulating the inflammatory response in mammals. The present study aimed to investigate the function of large yellow croaker FFAR4 on inflammation. In the present study,ffar4was widely expressed in 10 tissues of large yellow croaker including gill, head kidney and spleen. Further studies showed that treatment of head kidney macrophages with agonists (TUG891 or GSK137647A) or overexpression offfar4reduced the mRNA expression of pro-inflammatory genes induced by LPS, and increased the expression ofpparγ. Treatment of macrophages with antagonist AH7614 increased the mRNA expression of pro-inflammatory genes induced by LPS, and decreased the mRNA expression ofpparγ. In order to verify the immunomodulatory effect of PPARγ, PPARγ was overexpressed in macrophages which significantly reduced the mRNA expression of pro-inflammatory genesil6,il1β,il8,tnfαandcox2. Moreover, results of dual-luciferase assays showed that PPARγ downregulated the transcriptional activity ofil6andil1βpromoters. In conclusion, FFAR4 showed anti-inflammatory effects on LPS-induced inflammation in large yellow croaker.

A non-contact method was proposed to monitor the freshness (based on TVB-N and TBA values) of large yellow croaker fillets (Larimichthys crocea) by using a visible and near-infrared hyperspectral imaging system (400–1000 nm). In this work, the quantitative calibration models were built by using feed-forward neural networks (FNN) and partial least squares regression (PLSR). In addition, it was established that using a regression coefficient on the data can be further compressed by selecting optimal wavelengths (35 for TVB-N and 18 for TBA). The results validated that FNN has higher prediction accuracies than PLSR for both cases using full and selected reflectance spectra. Moreover, our FNN based model has showcased excellent performance even with selected reflectance spectra with rp = 0.978, R2p = 0.981, and RMSEP = 2.292 for TVB-N, and rp = 0.957, R2p = 0.916, and RMSEP = 0.341 for TBA, respectively. This optimal FNN model was then utilized for pixel-wise visualization maps of TVB-N and TBA contents in fillets.

The research evaluated the effects of locust bean gum (LBG) and sodium alginate (SA) active coatings containing 0.15, 0.30 or 0.60% lemon verbena (Lippa citriodora Kunth.) essential oil (LVEO) on the bacterial diversity and myofibrillar proteins (MPs) of large yellow croaker during refrigerated storage at 4 °C for 18 days. Variability in the dominant bacterial community in different samples on the 0, 9th and 18th day was observed. Pseudomonas and Shewanella were the two major genera identified during refrigerated storage. At the beginning, the richness of Pseudomonas was about 37.31% and increased for control (CK) samples during refrigerated storage, however, the LVEO-treated samples increased sharply from day 0 to the 9th day and then decreased. LBG-SA coatings containing LVEO treatments significantly delayed MPs oxidation by retarding the formation of free carbonyl compounds and maintaining higher sulfhydryl content, higher Ca2+-ATPase activity, better organized secondary (higher contents of α-helix and β-sheet) and tertiary structures during refrigerated storage. The transmission electron microscope (TEM) images showed that the integrity of the sarcomere was damaged, the boundaries of the H-, A-, and I-bands, Z-disk, and M-line were fuzzy in the CK samples at the end of storage. However, the LVEO-treated samples were still regular in appearance with distinct dark A-bands, light I-bands, and Z-disk. In brief, LBG-SA active coatings containing LVEO treatments suggested a feasible method for protecting the MPs of large yellow croaker during refrigerated storage.

At present, the methods for monitoring the health status of fish mostly utilize blood biochemical indicators. As the blood collection process often causes damage to the fish body (sometimes causing death), the need to develop a noninvasive detection method is urgent. Previous studies have shown that stress indicators can be detected in fish skin mucus or the surrounding water. In this experiment, the level of cortisol in the blood, fish skin mucus, and surrounding water of large yellow croaker was determined at 0, 0.5, 1, 1.5, and 2 h after high-temperature stress induction. Moreover, the levels of malondialdehyde (MDA), immunoglobulin M (IgM), and alkaline phosphatase (AKP) in the serum and fish skin mucus were also measured. The results were as follows: with increasing stress duration, the cortisol levels in the serum, fish skin mucus, and surrounding water increased first and then decreased, all reaching a peak at 1.5 h. The Pearson correlation coefficients for the cortisol levels between the serum and skin mucus, serum and surrounding water, and skin mucus and surrounding water were 0.936, 0.955, and 0.915, respectively, all with a significance level of P < 0.01. The MDA and IgM levels in the serum and fish skin mucus both increased first and then decreased, while the AKP levels in the serum and skin mucus decreased first and then increased. For the MDA, IgM, and AKP levels, the Pearson correlation coefficients between the serum and skin mucus were 0.586, 0.762, and 0.792, respectively, all with a significance level of P < 0.01. According to the results mentioned above, we can use the surrounding water or skin mucus as media to monitor early stress responses in large yellow croaker under high-temperature stress.

High-density single-nucleotide polymorphism (SNP) genotyping array is an essential tool for genetic analyses of animals and plants. Large yellow croaker (Larimichthys crocea) is one of the most commercially important marine fish species in China. Although plenty of SNPs have been identified in large yellow croaker, no high-throughput genotyping array is available. In this study, a high-throughput SNP array named NingXin-I with 600K SNPs was developed and evaluated. A set of 82 large yellow croakers were collected from different locations of China and re-sequenced. A total of 9.34M SNPs were identified by mapping sequence reads to the large yellow croaker reference genome. About 1.98M candidate SNPs were selected for further analyses by using criteria such as SNP quality score and conversion performance in the final array. Finally, 579.5K SNPs evenly distributed across the large yellow croaker genome with an average spacing of 1.19 kb were proceeded to array production. The performance of NingXin-I array was evaluated in 96 large yellow croaker individuals from five populations, and 83.38% SNPs on the array were polymorphic sites. A further test of the NingXin-I array in five closely related species in Sciaenidae identified 26.68-56.23% polymorphic SNP rate across species. A phylogenetic tree inferred by using the genotype data generated by NingXin-I confirmed the phylogenetic distance of the species in Sciaenidae. The performance of NingXin-I in large yellow croaker and the other species in Sciaenidae suggested high accuracy and broad application. The NingXin-I array should be valuable for quantitative genetic studies, such as genome-wide association studies (GWASs), high-density linkage map construction, haplotype analysis, and genome-based selection.

Polyunsaturated fatty acids (PUFAs) not only serve as essential nutrients but also function as modulators of the immune response in marine fish. However, their immunomodulatory mechanism is poorly understood given that the underlying regulation of the innate immune response in fish has not been fully elucidated. Hence, study of the innate immunity of fish could help elucidate the mechanism by which PUFAs affect the fish immune response. Here, we used combined transcriptome analysis and in vitro experimentation to study the mechanism of LPS-induced inflammation. Transcriptome profiling indicated that LPS elicited strong pro-inflammatory responses featuring high expression levels of pathogen recognition receptors (PRRs) and cytokines along with the activation of NF-κB and MAPK signaling pathways. The transcription factor p65 alone could increase the transcription of IL1β by binding to the promoter of IL1β, and this promoting effect disappeared after mutation or deletion of its binding sites. We then examined the effects of PUFAs on the levels of gene expression and the abundance of proteins of critical kinases associated with LPS-induced inflammation. We found that LA exerts pro-inflammatory response while ALA, EPA, and DHA induced anti-inflammatory effects by modulating the expression of PRRs, phosphorylation of IKK and p38, and the nuclear translocation of p65. Overall, this study advances our understanding of the regulatory mechanisms by which PUFAs regulate LPS-induced inflammation in a non-model fish species.

The p53 activation is induced by stressors, such as DNA damage, oxidative stress, and activated oncogenes, and can promote cell cycle arrest, cellular senescence, and apoptosis. The large yellow croaker (Larimichthys crocea) is an important warm temperate marine fish in the Chinese aquiculture industry. However, few studies have investigated the role of p53 in the response of L. crocea to environmental stressors. Therefore, the aim of the present study was to assess the spatiotemporal mRNA expression levels of genes involved in the p53 signaling pathway of the large yellow croaker in response to cold stress. The results showed significant changes in the expression levels of p53, p21, MDM2, IGF-1, Gadd45, Fas, and Akt in various tissues of the large yellow croaker in response to cold stress for different times. As compared to the control group, p53 mRNA expression was upregulated in most of the examined tissues at 24 h with the exception of the gill. In the liver, the expression levels of p53 and Fas were significantly decreased at 12 h, while those of p21, MDM2, IGF-1, Gadd45 were dramatically increased. Akt expression was notably changed in response to cold in several tissues. These results suggested that p53 was potentially a key gene in the large yellow croaker response to cold and possibly other environmental stressors.

Given the biogeochemical functions of marine microbes, the structure and functional diversity of bacterioplankton communities could be regarded as bioindicators that reflect environmental quality. The habitat of the large yellow croaker (Pseudosciaena crocea) is suffering from widespread degradation, but the study of the croaker habitat is still insufficient. Here, we used 16S rRNA gene sequencing and functional prediction to investigate the surface bacterioplankton communities in an overwintering ground of P. crocea during the overwintering period (November 2018 to April 2019). Our results showed that the taxa and functions of the environmental bacterioplankton community exhibited obvious seasonality. Specifically, the bacterioplankton communities in autumn were characterized by animal parasites or symbionts, such as Acinetobacter baumannii and Moraxella, while organic substance–degrading bacteria, such as Rhodococcus, Stenotrophomonas, and Alcanivorax, dominated in spring. Moreover, phylogenetic-based mean nearest taxon distance (MNTD) analyses indicated that dispersal limitation was the most important process governing the spatiotemporal assembly of the bacterioplankton communities. The distance decay of similarity also suggested the impact of dispersal limitation on the generation of biogeographical patterns. Furthermore, variation partitioning analysis (VPA) and a partial Mantel test revealed that environmental filters, such as nutrients, temperature, and salinity, had relatively weak deterministic effects on the bacterioplankton community assembly. Overall, these findings provide a basis for understanding the theory of surface microbial community assembly in the overwintering habitat of the large yellow croaker.

Interleukin-2 (IL-2) signals influence various lymphocyte subsets during differentiation, immune responses and homeostasis. IL-2 acts on different cells by binding to its receptors (IL-2R), which consists of three subunits, IL-2Rα (CD25), IL-2Rβ (CD122), and the common gamma chain or γc (CD132). In the present study, three IL-2 receptor subunits, designated as LcCD25-like (LcCD25L), LcIL-2Rβ and Lcγc, were characterized in large yellow croaker (Larimichthys crocea). The LcCD25L, like other teleost CD25L or IL-2/IL-15Rα, contains only one sushi domain at N-terminus. The synteny of CD25L from different teleost are conserved. The deduced protein of LcIL-2Rβ and Lcγc exhibits a typical class I cytokine receptors architecture, including a cytokine-binding homology domain (CHD) consisting of two fibronectin type-III (FNIII) domains (D1 and D2) and a conserved WSXWS motif in D2 domain. These three IL-2 receptor subunits were constitutively expressed in all tissues and primary immune-related cells examined. The LcCD25L was highly expressed in blood, while LcIL-2Rβ and Lcγc were highly expressed in spleen and gill. For immune-related cells, LcCD25L and LcIL-2Rβ were highly expressed in PKLs, while the Lcγc exhibited the highest expression in PKMs. These three IL-2 receptor subunits could be dramatically induced by T cell mitogen PHA in PKLs, which mainly composed of T and B lymphocytes. The results presented indicated that large yellow croaker IL-2R might exercise function on lymphocytes, especially on activated T cells.

Receptor-interacting protein 3 (RIP3) has been demonstrated to be a key regulator not only in cell death pathways including apoptosis and necroptosis but also in inflammation and host immune responses. In this study, a RIP3 ortholog named Lc-RIP3 is identified in large yellow croaker (Larimichthys crocea). The open reading frame (ORF) of Lc-RIP3 is 1524 bp long and encodes a protein of 507 amino acids (aa). The deduced Lc-RIP3 protein has an N-terminal kinase domain and a C-terminal RHIM domain, and the genome organization of Lc-RIP3 is conserved in teleosts with 12 exons and 11 introns but is different from that in mammals, which comprises 10 exons and 9 introns. Confocal microscopy revealed that Lc-RIP3 is a cytosolic protein. The expression analysis at the mRNA level indicated that Lc-RIP3 is ubiquitously distributed in various tissues/organs, and could be up-regulated under poly I:C, LPS, PGN, and Pseudomonas plecoglossicida stimulation in vivo. Notably, Lc-RIP3 could induce NF-κB but not IRF3 activation. In addition, Lc-RIP3 co-expression with Lc-TRIF, Lc-MAVS, or Lc-IRF3 significantly abolishes the activation of NF-κB but enhances the induction of IRF3 activity. Moreover, NF-κB activity could be up-regulated when Lc-RIP3 is co-expressed with Lc-RIP1 or Lc-IRF7. These results collectively indicate that Lc-RIP3 acts as an important regulator in host innate immune signaling in teleosts.

Large yellow croaker is an important marine aquaculture species in China. The aim was to determine an appropriate protocol of artificial fertilization for family construction in the breeding programme based on two trials. In trial 1, luteinizing hormone-releasing hormone A3 (LHRHA3) was injected once, with a dosage of 2 μg/kg for females and 1 μg/kg for males. The latency time was in the range of 30–34 h. The maturation stage was checked by extracting a few eggs with a Pasteur pipette. The fertilization rate and hatching rate were 27 and 52%, respectively. The percentage of females with spawning difficulties was 30%. In trial 2, the females were injected LHRHA3 twice: with a first dose of 0.8 μg/kg and a second dose of 2 μg/kg, at an interval of 10 h, whereas the males were still injected once. The latency time was in the range of 29.5–35 h, determined by only observing courtship behaviour of males. The females with spawning difficulties decreased to 10%, and the fertilization rate and hatching rate also improved to 41 and 62%, respectively.

Elongation of very long chain fatty acids protein 6 (Elovl6) is a key enzyme in fatty acid synthesis, which participates in converting palmitate (C16:0) to stearate (C18:0). Although studies of Elovl6 have been carried out in mammals, the nutritional regulation of elovl6 in fish remains poorly understood. In the present study, the cloning and nutritional regulation of elovl6 were determined in large yellow croaker. Sequence and phylogenetic analysis revealed that the full-length cDNA of elovl6 was 1360 bp, including an open reading frame of 810 bp encoding a putative protein of 269 amino acid that possesses the characteristic features of Elovl proteins. The transcript level of elovl6 was significantly increased in the liver of croaker fed the diets with soybean oil (enriched with 18: 2n-6, LA) or linseed oil (enriched with 18: 3n-3, ALA) than that in croaker fed the diet with fish oil (enriched with 20: 5n-3 and 22: 6n-3). Correspondingly, the elovl6 expression in croaker&rsquo, s hepatocytes treated with ALA or LA was remarkably increased compared to the controls. Furthermore, the transcription factors including hepatocyte nuclear factor 1&alpha, (HNF1&alpha, ), CCAAT-enhancer-binding protein &beta, (CEBP&beta, ), retinoid X receptor &alpha, (RXR&alpha, ), and cAMP response element-binding protein 1 (CREB1) greatly enhanced promoter activity of elovl6 in large yellow croaker, and the expression of transcription factors is consistent with the changes of elovl6 expression in response to fatty acids in vivo and in vitro. In conclusion, this study revealed that elovl6 expression in large yellow croaker could be upregulated by dietary ALA or LA via the increased transcriptional expression of transcription factors including hnf1&alpha, cebp&beta, rxr&alpha, and creb1.

Sansha Bay, facing the East China Sea, is characterised by high fish production (Pseudosciaena crocea, 91,000 t yr–1) and the polyculture of seaweeds (Saccharina japonica and Gracilaria lemaneiformis) and abalone (Haliotis discus hannai). To investigate the environmental effects of fish farming on sediments, surface (0–1 cm) sediments were collected from 41 stations grouped into four areas: fish, seaweed, and abalone farms and reference area. The physical and chemical parameters of these sediments were analysed. Nitrogen and phosphorus loadings from trash fish as feed were estimated to be 12,000 and 2,400 t yr–1, respectively, accounting for 90% and 86% of the total anthropogenic loading in Sansha Bay. However, no significant differences were observed in stable C and N isotope ratios, carbon-to-nitrogen ratio, total organic carbon, total nitrogen, total phosphorus, acid volatile sulfide, water content, and silt–clay fraction among the four areas (Kruskal–Wallis test, P > 0.05). Principal component and cluster analyses supported this conclusion, indicating that fish farming has no significant effect on the sediment. The amount of N removed by the cultured seaweed was estimated to be 288 t yr–1, accounting for only 2% of the N loaded from fish farming. The results suggested that the dispersion of fish farm-derived wastes were primarily facilitated by the topographic and hydrodynamic characteristics of Sansha Bay, such as its large depth (∼90 m), strong currents (1.4–1.9 m s–1), large tidal amplitude (5.4 m), and rapid exchange (

Large yellow croaker (LYC), Pseudosciaena crocea is an economically important fish species of mariculture in China. The variation of yearly production of LYC has been increasingly related to the outbreaks of fish diseases. Moreover, Vibrio infections have been identified in this fish frequently. To understand the pattern of Vibrio infections in LYC, we conducted a culture-independent survey of Vibrios in farmed LYC populations using a multiplex PCR method targeting Vibrio alginolyticus, Vibrio harveyi and Vibrio parahaemolyticus. The results showed that three fish pathogenic Vibrios had been detected in LYC populations at each sampling with a prevalence ranging from 6.7% to 73.3% but no single species dominated the Vibrio infection. The findings indicate that three Vibrio species still have impact on health status of farmed LYC and LYC aquaculture requires more efficacious prophylactic strategies. Keywords: Vibrio, Large yellow croaker, Multiplex PCR, Epidemiology

The present study aimed to investigate the macroscopic and histological structure of the gastrointestinal tract (GIT) of Larimichthyscrocea (Richardson, 1846). It consists of esophagus, stomach regions, pyloric caeca, intestinal regions, and rectum. Sixteen tubular light yellowish pyloric caeca of similar sizes were observed in all individuals. The digestive wall consists of mucosa, submucosa, muscularis, and adventitia. No major differences appeared in the structure of the tunica, epithelial cell types, connective tissues and musculature glands of L.crocea GIT. The mucosal epithelia in the oesophagus has longitudinal branched folds with frontward and hindmost zones. The gastric tunica mucosa has a characteristic folded structure and can be divided into three regions. The intestinal tunica mucosa is characterized by villi structures and numerous mucus-secreting cells. Mucus-secreting goblet cells were strongly positive to AB at pH 2.5 in the oesophagus (excluding gastro-oesophageal junction) and intestine mucosal regions, which indicates an abundance of carboxylate mucins. The surface epithelia of the gastric mucosa is PAS-positive and AB-negative. SEM examination revealed that cells in the epithelium of the esophagus have an unbroken apical layer and goblet cells. The intestinal coefficient (IC) of L.crocea was 0.80 ± 0.21, consistent with a carnivorous or omnivorous habit. Our study adds the knowledge of the digestive system of L.crocea and might be useful in the management of L.crocea stocks.

Heat shock proteins 70 (Hsp70) are required for key cellular processes and responses to environmental changes, however, there are an unknown number of hsp70 gene family members in the large yellow croaker (Larimichthys crocea). In the present study, 17 hsp70 genes were identified through the genome of the large yellow croaker. These genes are divided into seven evolutionarily distinct groups according to a phylogenetic tree. The orthologs of these hsp70 genes were found in humans and zebrafish. The expression patterns of the hsp70 gene family in the large yellow croaker under cold and heat stress were studied by examining transcriptome data. Six out of 17 genes were significantly unregulated or downregulated after cold or heat stress. There were two genes significantly upregulated and two genes downregulated in the liver after cold treatment, while after heat treatment, five genes were significantly upregulated, and no genes were significantly downregulated. Three expression patterns were detected: strictly heat-inducible hsp70, constitutively expressed and moderately heat-inducible hsp70, and constitutively expressed and less stress-dependent hsp70 genes. All the findings will contribute to a better understanding of the biological function of hsp70s in defending against thermal challenges.

Early life stages of marine organisms are the most sensitive stages to environment stressors including pollutants. In order to understand the toxicological effects induced by MeHg exposure on juveniles of large yellow croaker (Pseudosciaena crocea), a toxicity test was performed wherein fish were exposed to sub-lethal concentrations of MeHg under laboratory conditions (18 ± 1°C; 26 ± 1 in salinity). After 30 days of 0–4.0 μg L-1 MeHg exposure, SOD activity was significantly decreased in the 0.25, 1.0, and 4.0 μg L-1 treatments; while CAT activity was significantly increased in the 4.0 μg L-1 treatments; GSH level, GPx activity were significantly elevated in the 4.0 μg L-1 treatments, respectively. Meanwhile, malondialdehyde content was also significantly increased in the 1.0 and 4.0 μg L-1 treatments with respect to the control. Acetylcholinesterase activity was significantly decreased by 18.3, 25.2, and 21.7% in the 0.25, 1.0, and 4.0 μg L-1 treatments, respectively. The expression of TCTP, GST3, Hsp70, Hsp27 mRNA were all up-regulated in juveniles with a dose-dependent manner exposed to MeHg. These results suggest that large yellow croaker juveniles have the potential to regulate the levels of antioxidant enzymes and initiate immune response in order to protect fish to some extent from oxidative stress induced by MeHg.

The piscidin family, which includes potent antimicrobial peptides with broad-spectrum activity, plays an important role in the innate immune system of fish. In this study, we cloned piscidin-5-like type 3 (Lcpis5lt3) in large yellow croaker (Larimichthys crocea). Multiple alignments with other known piscidins revealed amino acid conservation throughout the fish, especially at the signal peptide (22 amino acids). The phylogenetic tree confirmed that Lcpis5lt3 and large yellow croaker piscidin-5-like proteins were grouped together to form a branch. Quantitative real-time PCR revealed that Lcpis5lt3 was expressed in a wide range of tissues, including the brain, muscle, gill, head kidney, intestine, kidney, liver, and spleen. The highest mRNA expression level of Lcpis5lt3 was found in the spleen. After Vibrio alginolyticus infection, mRNA expression was rapidly upregulated in the liver, head kidney, gill, kidney, and intestine at 4, 8, 12, and 24 h post infection (hpi), whereas there were no significant changes in the spleen. The antimicrobial spectrum showed that the synthetic mature peptide of Lcpis5lt3 exhibited different activity in vitro against various bacteria, such as Aeromonas hydrophila, V. anguillarum, V. alginolyticus, V. parahaemolyticus, Staphylococcus aureus, and Listeria monocytogenes. In addition, survival rates from the in vivo assay indicated that the synthetic peptide of Lcpis5lt3 increased the survival rate of large yellow croaker after V. alginolyticus challenge, resulting in a decline in bacterial burden and mRNA expression levels of interleukin-1β, interleukin-10, and tumor necrosis factor-α. These data suggest that Lcpis5lt3 plays an important role in innate immunity in large yellow croaker and might represent a potential therapeutic agent against pathogen invasion.

Whole-genome single-nucleotide polymorphism (SNP) markers are valuable genetic resources for the association and conservation studies. Genome-wide SNP development in many teleost species are still challenging because of the genome complexity and the cost of re-sequencing. Genotyping-By-Sequencing (GBS) provided an efficient reduced representative method to squeeze cost for SNP detection; however, most of recent GBS applications were reported on plant organisms. In this work, we used anEcoRI-NlaIII based GBS protocol to teleost large yellow croaker, an important commercial fish in China and East-Asia, and reported the first whole-genome SNP development for the species. 69,845 high quality SNP markers that evenly distributed along genome were detected in at least 80% of 500 individuals. Nearly 95% randomly selected genotypes were successfully validated by Sequenom MassARRAY assay. The association studies with the muscle eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) content discovered 39 significant SNP markers, contributing as high up to ∼63% genetic variance that explained by all markers. Functional genes that involved in fat digestion and absorption pathway were identified, such asAPOB,CRATandOSBPL10. Notably,PPT2Gene, previously identified in the association study of the plasma n-3 and n-6 polyunsaturated fatty acid level in human, was re-discovered in large yellow croaker. Our study verified thatEcoRI-NlaIII based GBS could produce quality SNP markers in a cost-efficient manner in teleost genome. The developed SNP markers and the EPA and DHA associated SNP loci provided invaluable resources for the population structure, conservation genetics and genomic selection of large yellow croaker and other fish organisms.

Large yellow croaker (LYC) cage farming is a rapidly developing industry in the coastal areas of the East China Sea. However, little is known about the environmental nutrient loadings resulting from the current aquaculture practices for this species. In this study, a nitrogenous waste model was developed for LYC based on thermal growth and bioenergetic theories. The growth model produced a good fit with the measured data of the growth trajectory of the fish. The total, dissolved and particulate nitrogen outputs were estimated to be 133, 51 and 82 kg N tonne(-1) of fish production, respectively, with daily dissolved and particulate nitrogen outputs varying from 69 to 104 and 106 to 181 mg N fish(-1), respectively, during the 2012 operational cycle. Greater than 80 % of the nitrogen input from feed was predicted to be lost to the environment, resulting in low nitrogen retention (20 %) in the fish tissues. Ammonia contributed the greatest proportion (85 %) of the dissolved nitrogen generated from cage farming. This nitrogen loading assessment model is the first to address nitrogenous output from LYC farming and could be a valuable tool to examine the effects of management and feeding practices on waste from cage farming. The application of this model could help improve the scientific understanding of offshore fish farming systems. Furthermore, the model predicts that a 63 % reduction in nitrogenous waste production could be achieved by switching from the use of trash fish for feed to the use of pelleted feed.

The importance of a gene’s impact on traits is well appreciated. Gene expression will affect the growth, immunity, reproduction and environmental resistance of some fish, and then affect the economic performance of fish-related business. Studying the connection between gene and character can help elucidate the growth of fishes. Thus far, a collected database containing large yellow croaker (Larimichthys crocea) genes does not exist. The gene having to do with the growth efficiency of fish will have a huge impact on research. For example, the protein encoded by the IFIH1 gene is associated with the function of viral infection in the immune system, which affects the survival rate of large yellow croakers. Thus, we collected data through the published literature and combined them with a biological genetic database related to the large yellow croaker. Based on the data, we can predict new gene–trait associations which have not yet been discovered. This work will contribute to research on the growth of large yellow croakers.

The therapeutic effect of polysaccharide of large yellow croaker swim bladder (PLYCSB) on lupus nephritis has been studied in vivo. A high concentration (50 mg/kg dose) of PLYCSB reduced the levels of serum inflammatory cytokine levels of IL-6, IL-12, TNF-α and IFN-γ compared to a low concentration (25 mg/kg dose) and control mice. SCr, BUN, TC and TG serum levels of PLYCSB treated mice were lower than those of control mice, and TP and ALB serum levels were higher than control mice. Control mice tested ds-DNA positive at the 6th week, and 50 mg/kg treated mice tested at the 10th week after the experiment began. The output of urine protein of 50 mg/kg PLYCSB treated mice was most closely comparable to the normal mice. The glomerular number of 50 mg/kg PLYCSB treated mice was more than the 25 mg/kg dose and control groups, and the 50 mg/kg dose group showed the lowest glomerular sclerosis index in lupus nephritis mice. By RT-PCR and western blot assay, PLYCSB significantly induced inflammation in kidney tissues of mice by downregulating NF-κB-p65, TGF-β1, Fas, FasL and upregulating IκB-α. These results suggest that PLYCSB showed a potential curative effect on lupus nephritis as a drug or functional food.

International audience; A comprehensive survey of channel catfish Toll-like receptors (TLRs) was undertaken following a genomic PCR approach based on degenerate primers. Twenty different TLRs were identified in channel catfish. Channel catfish TLR sequences were characterized by phylogenetic analysis based on their conserved Toll/interleukin-1 receptor domain and by in-depth analysis of leucine-rich repeat (LRR) motifs of the ligand binding extracellular domain (ECD). The catfish have representatives of all the TLR types defined in vertebrates with the exception of TLR6, TLR10, TLR11, TLR12, TLR13, TLR15, TLR23, and TLR24. Additionally, two new types were discovered: TLR25 and TLR26. TLR25 is also present in cyprinids, cichlids, plecoglossids, and adrianichthyids, suggesting its presence early in fish evolution. To date, TLR26 was found only in channel catfish. Like TLR18-23, TLR25 and TLR26 were not found in any other vertebrate classes and appear to be fish specific. Data mining using the catfish TLR sequences revealed that in addition to ictalurids and cyprinids, TLR4 is also present in salmonids. TLR19 and TLR20 were both found in ictalurids, cyprinids, and salmonids, demonstrating a wider range than previously known. The LRR structure within ECDs appeared generally well conserved. TLR7 demonstrated a very high identity to human TLR7 strongly suggesting that ligand specificity maybe conserved. Finally, expression profiling confirmed that most TLRs are widely expressed in a diversity of tissues and revealed marked differences of expression level.

Little research has been done in optimizing the nitrogenous fraction of the fish diets in order to minimize welfare problems. The purpose of this review is to give an overview on how amino acid (AA) metabolism may be affected when fish are under stress and the possible effects on fish welfare when sub-optimal dietary nitrogen formulations are used to feed fish. In addition, it intends to evaluate the current possibilities, and future prospects, of using improved dietary nitrogen formulations to help fish coping with predictable stressful periods. Both metabolomic and genomic evidence show that stressful husbandry conditions affect AA metabolism in fish and may bring an increase in the requirement of indispensable AA. Supplementation in arginine and leucine, but also eventually in lysine, methionine, threonine and glutamine, may have an important role in enhancing the innate immune system. Tryptophan, as precursor for serotonin, modulates aggressive behaviour and feed intake in fish. Bioactive peptides may bring important advances in immunocompetence, disease control and other aspects of welfare of cultured fish. Fishmeal replacement may reduce immune competence, and the full nutritional potential of plant-protein ingredients is attained only after the removal or inactivation of some antinutritional factors. This review shows that AA metabolism is affected when fish are under stress, and this together with sub-optimal dietary nitrogen formulations may affect fish welfare. Furthermore, improved dietary nitrogen formulations may help fish coping with predictable stressful events.

The ontogenetic development of the digestive tract and accessory organs in large yellow croaker (Pseudoscianea crocea) was described using light microscopy from hatching up to the juvenile stage (40 days post hatch, dph). At 3 dph (4.1 � 0.1 mm total length, LT), coinciding with the buccopharynx opening, larvae started to feed exogenously, and the digestive tract consisted of a well- developed buccopharynx, a partially-differentiated oesophagus, and an intestine divided in three regions (prevalvular and postvalvular intestine, and rectum). Yolk reserves were not completely depleted at the onset of exogenous feeding, and a period of mixed nutrition was observed up to 6 dph (4.3 � 0.1 mm LT), when yolk was definitively exhausted. Important morphological changes occurred at the end of the larval period, coinciding with metamorphosis. At 17 dph (6.8 � 0.6 mm LT), pyloric caeca differentiated at the junction of the pyloric stomach and the prevalvular intestine. Gastric glands were firstly observed at 21 dph (9.2 � 1.2 mm LT), coinciding with the morphological development of the stomach in three different regions (cardiac, fundic and pyloric) according to the histological characteristics of their mucosa. At this age, large longitudinal folds appeared in the median and posterior oesophageal mucosa. These morphological and histological features suggested the achievement of a digestive system characteristic of large yellow croaker juveniles and adults.

Background Large yellow croaker (Larimichthys crocea) is an important commercial fish in China and East-Asia. The annual product of the species from the aqua-farming industry is about 90 thousand tons. In spite of its economic importance, genetic studies of economic traits and genomic selections of the species are hindered by the lack of genomic resources. Specifically, a whole-genome physical map of large yellow croaker is still missing. The traditional BAC-based fingerprint method is extremely time- and labour-consuming. Here we report the first genome map construction using the high-throughput whole-genome mapping technique by nanochannel arrays in BioNano Genomics Irys system. Results For an optimal marker density of ~10 per 100 kb, the nicking endonuclease Nt.BspQ1 was chosen for the genome map generation. 645,305 DNA molecules with a total length of ~112 Gb were labelled and detected, covering more than 160X of the large yellow croaker genome. Employing IrysView package and signature patterns in raw DNA molecules, a whole-genome map of large yellow croaker was assembled into 686 maps with a total length of 727 Mb, which was consistent with the estimated genome size. The N50 length of the whole-genome map, including 126 maps, was up to 1.7 Mb. The excellent hybrid alignment with large yellow croaker draft genome validated the consensus genome map assembly and highlighted a promising application of whole-genome mapping on draft genome sequence super-scaffolding. The genome map data of large yellow croaker are accessible on lycgenomics.jmu.edu.cn/pm. Conclusion Using the state-of-the-art whole-genome mapping technique in Irys system, the first whole-genome map for large yellow croaker has been constructed and thus highly facilitates the ongoing genomic and evolutionary studies for the species. To our knowledge, this is the first public report on genome map construction by the whole-genome mapping for aquatic-organisms. Our study demonstrates a promising application of the whole-genome mapping on genome maps construction for other non-model organisms in a fast and reliable manner. Electronic supplementary material The online version of this article (doi:10.1186/s12864-015-1871-z) contains supplementary material, which is available to authorized users.

Background The advances of sequencing technology accelerate the development of theory of molecular quantitative genetics such as QTL mapping, genome-wide association study and genomic selection. This paper was designed to study genomic selection in large yellow croaker breeding. The aims of this study were: (i) to estimate heritability values of traits in large yellow croaker; (ii) to assess feasibility of genomic selection in the traits of growth rate and meat quality; (iii) to compare predictive accuracies affected by different algorithms and training sizes, and to find what training sizes could reach ideal accuracies; (iv) to compare results of GWAS with genomic prediction, and to assess feasibility of pre-selection of significant SNPs in genomic selection. 500 individuals were tested in the trait of body weight and body length, while 176 were tested in the percentage of n-3 highly unsaturated fatty acids (n-3HUFA) in muscle. GBLUP and emBayesB were used to perform genomic prediction. Results Genotyping-By-Sequencing method was used to construct the libraries for the NGS sequencing and find ~30,000 SNPs. Heritability estimates were 0.604, 0.586 and 0.438 for trait of body weight, body length and n-3HUFA, respectively. The predictive abilities estimated by GBLUP showed higher than that by emBayesB in traits of body weight and body length. However, the result was just the opposite in n-3HUFA. According to fit the curve of predictive accuracy, we estimated that at least 1000 individuals in training set could reach an accuracy of 0.8 in body weight and body length. GBLUP, emBayesB and GWAS could not always find significant SNPs associated with phenotypes consistently. Significant SNPs were selected by emBayesB could obtain the largest proportions to explain total additive genetic variances. Conclusions This research showed that genomic selection was feasible in large yellow croaker breeding. We suggest doing a test before deciding to use which algorithm in specific trait in genomic prediction. We estimated required training sizes to reach ideal predictive accuracies and assessed feasibility of pre-selection of SNPs successfully. Because of high mortality rate of fish and high cost in genomic sequencing, genomic selection may be more suitable for applying on some traits which cannot be measured on candidates directly.