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To probe the mechanisms of gill remodeling in blunt snout bream under hypoxic conditions, we selected gill tissue for primary cell culture to establish and characterize the first blunt snout bream gill cell line, named MAG. The gill cells were efficiently passaged in M199 medium supplemented with 8% antibiotics and 15% fetal bovine serum at 28 °C, exhibiting primarily an epithelial-fibroblast mixed type. Additionally, the MAG cells (17th generation) were subjected to four experimental conditions—normoxia, hypoxia 12 h, hypoxia 24 h, and reoxygenation 24 h (R24h)—to evaluate the effects of hypoxia and reoxygenation on MAG cells during gill remodeling. We found that the MAG cell morphology underwent shrinkage and mitochondrial potential gradually lost, even leading to gradual apoptosis with increasing hypoxia duration and increased reactive oxygen species (ROS) activity. Upon reoxygenation, MAG cells gradually regain cellular homeostasis, accompanied by a decrease in ROS activity. Analysis of superoxide dismutase (SOD), glutathione (GSH), lactate dehydrogenase (LDH), catalase (CAT), anti-superoxide anion, and other enzyme activities revealed enhanced antioxidant enzyme activity in MAG cells during hypoxia, aiding in adapting to hypoxic stress and preserving cell morphology. After reoxygenation, the cells gradually returned to normoxic levels. Our findings underscore the MAG cells can be used to study hypoxic cell apoptosis during gill remodeling. Therefore, the MAG cell line will serve as a vital in vitro model for exploring gill remodeling in blunt snout bream under hypoxia. [ABSTRACT FROM AUTHOR]

This study investigated the effect of oral bovine serum albumin (BSA) on plasma amino acid profile and growth performance in blunt snout bream fed high‐fat and high‐energy diets. A total of 360 fish (average weight: 45.84 ± 0.07 g) were randomly divided into three groups in four replicates and fed with the control, high‐fat diet (HFD), and high‐energy diet (HED), respectively. After 12 weeks of feeding, fish were orally administered with 10% BSA for 10 h, with the blood and liver sample obtained at 10 h. The results showed no remarkable difference (p > 0.05) in final weight, weight gain rate, feed conversion ratio, protein efficiency ratio, and daily growth rate among dietary treatments. However, fish fed HFD and HED obtained significantly higher energy and nitrogen retention content than the control. Plasma valine and histidine content were more prevalent in fish fed the control diet than in HFD and HED groups. However, BSA significantly enhanced this content with higher values observed in arginine and alanine content than in control diets at 10 h. Significantly higher liver and plasma Aspartate aminotransferase, Xanthine oxidase, and Glutamate dehydrogenase content were found in the HFD than in the control group. However, the administration of BSA at 10 h improved these parameters similar to the control. Meanwhile, protease, Lipase, aminopeptidase N, carboxypeptidase A, and Na+ K+‐ATPase were influenced significantly (p < 0.001) by sampling time, dietary treatments, and their interaction. Fish fed with the HFD and HED significantly downregulated (p < 0.05) the akt, bcat2, 4e‐bp1, s6k1, and tor expression compared to the control. However, BSA administration at 10 h improved these genes similar to the control. This study indicated that oral BSA administration could benefit plasma amino acid through the TOR signaling pathway in blunt snout bream fed high‐fat and high‐energy diets. [ABSTRACT FROM AUTHOR]

Abstract This study investigated the effect of oral bovine serum albumin (BSA) on plasma amino acid profile and growth performance in blunt snout bream fed high‐fat and high‐energy diets. A total of 360 fish (average weight: 45.84 ± 0.07 g) were randomly divided into three groups in four replicates and fed with the control, high‐fat diet (HFD), and high‐energy diet (HED), respectively. After 12 weeks of feeding, fish were orally administered with 10% BSA for 10 h, with the blood and liver sample obtained at 10 h. The results showed no remarkable difference (p > 0.05) in final weight, weight gain rate, feed conversion ratio, protein efficiency ratio, and daily growth rate among dietary treatments. However, fish fed HFD and HED obtained significantly higher energy and nitrogen retention content than the control. Plasma valine and histidine content were more prevalent in fish fed the control diet than in HFD and HED groups. However, BSA significantly enhanced this content with higher values observed in arginine and alanine content than in control diets at 10 h. Significantly higher liver and plasma Aspartate aminotransferase, Xanthine oxidase, and Glutamate dehydrogenase content were found in the HFD than in the control group. However, the administration of BSA at 10 h improved these parameters similar to the control. Meanwhile, protease, Lipase, aminopeptidase N, carboxypeptidase A, and Na+ K+‐ATPase were influenced significantly (p

The blunt snout bream, Megalobrama amblycephala, is a commercially significant freshwater fish species indigenous to China. In recent decades, fishery resources of blunt snout bream in the Yangtze River have been severely depleted owing to human activities, and limited information is available regarding their genetic characteristics. This study aimed to assess the genetic diversity and population structure of different native populations of blunt snout bream in the Yangtze River. We analyzed 222 samples from six natural populations in the middle reaches of the Yangtze River and its affiliated lakes were analyzed using ten microsatellite DNA markers and concatenated sequences of two mitochondrial DNA markers (Cytb and ND2). Microsatellite analysis showed that the average observed heterozygosity (Ho), expected heterozygosity (He), and polymorphic information content (PIC) for all populations were determined as 0.687, 0.739, and 0.748 respectively. Mitochondrial sequence analysis showed that the number of haplotypes (h), haplotype diversity (Hd) and nucleotide diversity (π) of the six populations were 29, 0.768, and 0.00641, respectively. The above findings reveal high genetic diversity within blunt snout bream populations, suggesting a robust potential for selective breeding. The phylogenetic relationship results showed that the six blunt snout bream populations clustered into two branches, indicating a measurable degree of differentiation among the populations in the middle reaches of the Yangtze River Basin and its affiliated lakes. The populations may represent distinct spawning grounds, and corresponding conservation units should be implemented to protect them. In summary, these findings provide vital genetic information for the conservation and commercialization of blunt snout bream in the Yangtze River Basin of China.

The blunt snout bream (Megalobrama amblycephala) is a typical hypoxia-sensitive fish, and hypoxia stress leads to reduced vitality and yield during aquaculture. To explore the specific adaptation mechanism under hypoxia, the blunt snout bream was treated with hypoxia (DO = 2.0 ± 0.1 mg/L) for 24 h, followed by 3 h of recovery. Our results depicted that the gill filament structure of blunt snout bream changed after hypoxia. During hypoxia for 24 h, the gill filament structure was altered, including a more than 80% expansion of the lamellar respiratory surface area and a proportionate apoptosis decrease in interlamellar cell mass (ILCM) volume. Thus, the water–blood diffusion distance was shortened to less than 46%. During hypoxia for 24 h, the activity of ROS in gill tissue increased significantly (p < 0.05), while the mitochondrial membrane potential decreased significantly (p < 0.05). During hypoxia, mRNA expression level of anti-apoptotic gene Bcl-2 in the gills of blunt snout bream decreased significantly (p < 0.05), while the expression of pro-apoptotic gene Bax mRNA increased significantly (p < 0.05). Thus, the ratio of Bax/Bcl-2 mRNA increased in the gills of blunt snout bream to promote the activity of Caspase-3. Together, our results indicated hypoxia-induced apoptosis in the gills of blunt snout bream through the mitochondrial pathway. In addition, a decreased expression of Phd1 and an increased expression of Hif-1α in gills under hypoxia stress indicates that blunt snout bream may cope with hypoxia-induced apoptosis by enhancing the HIF pathway. These results provide new insights into fish's adaptation strategies and mechanisms of hypoxia. [ABSTRACT FROM AUTHOR]

Ammonia–nitrogen is a common stress factor for aquatic organisms in their habitation environment, which is enriched in water due to high-density farming and environmental pollution. Ammonia nitrogen can enter fish body through gill, epidermis, digestive tract, and other tissues, causing fish ammonia poisoning. In the present study, juvenile blunt snout bream (average weight, 45 ± 5 g) were exposed to high concentrations of ammonia–nitrogen stress (25.0 ± 0.5 mg/L) for six different treatment times (0, 3, 6, 12, 24, 48, and 72 h); the tissue ultrastructure, mRNA levels of antioxidation system, and apoptosis patterns were studied. The antioxidant systems of malondialdehyde (MDA), catalase (CAT), acid phosphatase (ACP), and reduced glutathione (GSH) in various tissues were highly transcripted at 6 or 12 h (hpt) after treatment under high ammonia–nitrogen, which may play a role in preventing cells from being attacked by highly toxic reactive oxygen species (ROS). After 24 hpt, the antioxidant capacity threshold is breached, followed by the decline of antioxidant enzyme activity. Thus, with the prolonging of high ammonia–nitrogen processing time, ammonia–nitrogen stress caused irreversible damage of organs (gill, liver, and kidney). Furthermore, the expression of caspase-3 apoptotic pathway was highly induced in different tissues, implying the apoptotic system is activated, which causes extensive cell apoptosis in different tissues as shown using TUNEL analysis. In conclusion, we observed that, in response to acute ammonia–nitrogen stress, blunt snout bream enhances antioxidant capacity and cell apoptosis. [ABSTRACT FROM AUTHOR]

Based on the distant hybridization derived from Hypophthalmichthys nobilis (bighead carp, BIC, 2n ​= ​48) (♀) ​× ​Megalobrama amblycephala (blunt snout bream, BSB, 2n ​= ​48) (♂), we obtained a novel hybrid (BB) in this study. We comparatively investigated the morphological traits, chromosome numbers, DNA contents, microsatellite patterns, and DNA variations in 5S rDNA and Hox genes. The chromosome and flow cytometer results indicated that BB was a diploid hybrid with 48 chromosomes. We confirmed successful hybridization with microsatellite DNA markers and analyzed the Hox gene family in the parental and hybrid lines. The sequences of 5S rDNA in BB mapped to those in BIC and BSB, indicating that the genetic materials of BB were derived from BIC and BSB. The breeding experiment showed that BB had the characteristics of herbivore and fast growth. This diploid hybrid is of great significance for fish genetic breeding and applied research.

Interferons (IFNs) represent a group of proteins with notable antiviral capabilities. Recently, the discovery of type IV IFN, i.e. IFN-υ has created a new avenue for research on class II cytokines. In the present study, IFN-υ was identified in blunt snout bream to possess conserved sequence CXXXXX[W/L] and gene locus. In the bream, IFN-υ was expressed in examined tissues and was induced significantly following poly(I:C) stimulation. With the stimulation of recombinant IFN-υ in vivo, transcriptome was sequenced in head kidney, and 814 DEGs were identified with 165 possible IFN-stimulated genes (ISGs) characterized through comparison with type I IFN induced ISGs in zebrafish. Furthermore, IFN-υ was found to show high affinity with CRFB12, but no interaction was observed between IFN-υ and CRFB4, through surface plasmon resonance (SPR) and coimmunoprecipitation (Co-IP) analyses. These results should contribute to the understanding of IFN-υ and its receptor complex, and importantly its immune function in inducing ISGs. • IFN-υ was cloned in blunt snout bream with conserved features. • The type IV IFN was constitutively expressed and was induced by poly(I:C). • Transcriptome induced by recombinant IFN-υ contains about 170 ISGs. • IFN-υ was found to interact with CRFB12, but not CRFB4. [ABSTRACT FROM AUTHOR]

The blunt snout bream (Megalobrama amblycephala , BSB) is one of important freshwater fish of value in China, but it has poor tolerance to hypoxia environment. To improve this disadvantage, we cultivate an improved blunt snout bream (BTBB) from a hybrid lineage of Megalobrama amblycephala (♀) × Culter alburnus (♂). In this study, we found that the BTBB had higher tolerance hypoxia than BSB. Moreover, in order to take a deep insight into the molecular mechanisms of higher hypoxia tolerance in BTBB, we analyzed the liver transcriptomes of BTBB and BSB under normoxic and hypoxia conditions comparatively and found out several differentially expressed genes (DEGs). Among the GO functional categories, negative regulation of response to stimulus (GO: 0048585), oxidation-reduction process (GO: 0055114), and response to oxidative stress (GO: 0006979) in biological processes were observed. In addition, according to Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, the results indicated that many DEGs participated in FoxO signaling pathway, MAPK signaling pathway, VEGF signaling pathway, PPAR signaling pathway and p53 signaling pathway, and other processes. This study provides important clues about the potential molecular mechanisms explaining the higher hypoxia tolerance in BTBB. It also presents further evidence for the enhanced ability to resist hypoxia in BTBB, which is significant for fish genetic breeding. • The BTBB had higher tolerance hypoxia than BSB. • KEGG pathway analysis showed that many DEGs were enriched in the hypoxia response pathway. • The molecular mechanism of hypoxia tolerance of BTBB was analyzed. [ABSTRACT FROM AUTHOR]

To provide scientific support for improving the muscle quality of blunt snout bream, ultrahigh performance liquid chromatography, texture analyzer, and optical electron microscopy were applied to explore the effects of respite time (0, 24, 48, and 72 h) on the muscle quality of blunt snout bream before live transportation. The energy compounds (ATP and glycogen) of muscle significantly decreased with the respite time (p < 0.05). Lactic acid content declined and then increased, leading to a rise and then a fall in pH (p < 0.05). Water-holding capacity of fish muscle increased progressively when the respite time was increased to 48 h and then dramatically decreased when the respite time was further increased to 72 h (p < 0.05). Shear force gradually increased (p < 0.05), while the whiteness and lightness values remained stable (p > 0.05). Both the content of umami compounds (IMP) and bitter compounds (HxR, Hx), and the calculated K value decreased steadily with the respite time (p < 0.05). The extracellular gap of the fish muscles gradually decreased with the respite time. The most uniform and intact cellular structure occurred at 48 h. However, when the respite time was extended to 72 h, the extracellular gap and muscle fragmentation rate of the muscle increased considerably. The findings indicated that a 48 h respite time was suitable to improve the muscle quality of blunt snout bream after live transportation. [ABSTRACT FROM AUTHOR]

The surprising variation in the number of Hox clusters and genome structure in ray-finned fish lineages reflects the history of duplications and subsequent lineage-specific gene loss. However, there are few studies on whether Hox clusters in the early generations of hybrid lineages show more significant variation due to the continuous genomic oscillation caused by distant hybridization. We sequenced and analyzed Hox gene clusters from a crucian carp-like homodiploid fish (NCRC) lineage (a new hybrid lineage derived from common carp (Cyprinus carpio) (♀) ​× ​blunt snout bream (Megalobrama amblycephala) (♂)). In the NCRC lineage, we reconstructed seven Hox clusters consisting of 48 Hox genes, ten of which were pseudogenes. The number of putative Hox clusters generated in NCRC-F1 was increased greatly by distant hybridization to an average number almost twice that in the maternal parent. This increasing trend continued in the subsequent self-mating generations of NCRC-F1. In contrast, the number of Hox cluster fragments inherited from the original parents gradually decreased as the number of NCRC lineage generations increased. This pattern was also found in the inheritance of recombinant Hox clusters. In terms of base composition, some genetic rules for the inheritance of these Hox clusters between different generations of the NCRC lineage were identified. Furthermore, the newly derived mutated Hox clusters in the NCRC lineage showed phylogenetic relationships that were closer to either crucian carp or silver crucian carp, revealing a clear evolutionary path. This study deepens our understanding of the evolution of Hox genes in the ray-finned fish clade.

The present study was performed to determine the effects of Aeromonas hydrophila infection on intestinal -histopathology, innate immune response and changes in antioxidant capacity of blunt snout bream (Megalobrama amblycephala). A series of histopathological changes, innate immune enzyme activities, antioxidant enzyme activities, and the corresponding mRNA relative genes expressions in intestines were measured at 0, 1, 2, and 3 weeks post-treatment of Aeromonas hydrophila (1✕107 CFU mL−1) infection. The results showed that Aeromonas hydrophila induced changes in intestinal morphology, including the decreased muscularis thickness, the proliferated goblet cells, and the atrophied intestine villi height. Moreover, the innate immune enzymes activities in serum such as acid phosphatase, alkaline phosphatase, lysozyme activities and immunoglobulin M were significantly reduced after infection at 1week, 2week and 3week. The contents of complement 3 and complement 4 were significantly decreased after infection as well. In addition, the antioxidant enzymes activities, including superoxide dismutase, catalase and glutathione peroxidase in the experimental groups were significantly decreased compared with the control group, whereas the content of malondialdehyde was significantly increased after infection at 1week, 2week and 3week. Furthermore, the mRNA relative expressions of the inflammatory cytokines such as tumor necrosis factor-α, interleukins-1β, interferon-γ, and interleukins-6 were significantly increased after infection with Aeromonas hydrophila. The TJ-related gene expressions in the intestine of zonula occluden-1 , occludin , occludin-1 , occludin-2 were significantly reduced throughout the infection period. The mRNA relative expressions of signal transducers and activators of transcription 4 and janus kinase-3 in the intestine were significantly ascended compared with the non-infected group. Overall, the results elucidated that the intestine tissue injury and innate immune response reduction, as well as antioxidant capacity attenuation were occurred against Aeromonas hydrophila infection of the blunt snout bream. • Aeromonas hydrophila induced intestinal tissue damage of blunt snout bream. • Aeromonas hydrophila decreased innate immune response of blunt snout bream. • Aeromonas hydrophila declined antioxidant capacity of blunt snout bream. • Aeromonas hydrophila damaged the intestinal barrier immune function by regulating the JAK/STAT signaling pathway. [ABSTRACT FROM AUTHOR]

Blunt snout bream plays an important role in freshwater aquaculture in China, but the development of its culture industry has been restricted by increasing hypoxia problem. Through the breeding of wild blunt snout bream populations (F0), a hypoxia-tolerant new variety (F6) was obtained. In this study, the new variety was stressed under low oxygen concentration (2.0 mg·L−1) for 4 and 7 days, the morphological structure of the gill tissue showed a striking change, the interlamellar cell mass (ILCM) volume reduced significantly (P < 0.05), and the lamellar respiratory surface area enlarged significantly (P < 0.05), compared to normoxic controls. After 7 days of oxygen recovery, gill remodeling was completely reversed. Additionally, the TUNEL-positive apoptotic fluorescence signals increased in the gills exposed to hypoxia up to 4 and 7 days; the apoptosis rate also increased significantly (P < 0.05). Under 4 and 7 days of hypoxia stress, the expression of anti-apoptotic gene Bcl-2 in the gills downregulated significantly (P < 0.05), with the significantly (P < 0.05) upregulated expression of pro-apoptotic gene Bad. Furthermore, under hypoxia stress, the activity or content of oxidative stress–related enzymes (superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and glutathione (GSH)) in gill tissue increased to varying degrees compared to normoxic controls. These results offer a new perspective into the cellular and molecular mechanism of hypoxia-induced gill remodeling in blunt snout bream and a theoretical basis for its hypoxia adaptation mechanism. [ABSTRACT FROM AUTHOR]

To explore the gut microbial profile of blunt snout bream (Megalobrama amblycephala), fish were fed with a commercial diet for 16 weeks. Then high-throughput sequencing was applied to compare the microbiota between foregut and hindgut. The results showed that the microbial profiles between the foregut and hindgut were different based on the alpha-diversity and the cluster results. Dominant microbioorganisms of blunt snout bream at genus level were Cetobacterium spp., Lactococcus spp., CK-1C4-19, Rhodobacter spp. and Pseudomonas spp. Meanwhile it was found out that Lactococcus spp. preferred the foregut, while Cetobacterium spp. and Flavobacterium spp. preferred the hindgut. Dominant bacteria preferred different gut section may decided by its function and physiological characteristics. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Hydrobiology / Shuisheng Shengwu Xuebao is the property of Editorial Department of Journal of Hydrobiology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

To provide scientific support for improving the muscle quality of blunt snout bream, ultrahigh performance liquid chromatography, texture analyzer, and optical electron microscopy were applied to explore the effects of respite time (0, 24, 48, and 72 h) on the muscle quality of blunt snout bream before live transportation. The energy compounds (ATP and glycogen) of muscle significantly decreased with the respite time (p < 0.05). Lactic acid content declined and then increased, leading to a rise and then a fall in pH (p < 0.05). Water-holding capacity of fish muscle increased progressively when the respite time was increased to 48 h and then dramatically decreased when the respite time was further increased to 72 h (p < 0.05). Shear force gradually increased (p < 0.05), while the whiteness and lightness values remained stable (p > 0.05). Both the content of umami compounds (IMP) and bitter compounds (HxR, Hx), and the calculated K value decreased steadily with the respite time (p < 0.05). The extracellular gap of the fish muscles gradually decreased with the respite time. The most uniform and intact cellular structure occurred at 48 h. However, when the respite time was extended to 72 h, the extracellular gap and muscle fragmentation rate of the muscle increased considerably. The findings indicated that a 48 h respite time was suitable to improve the muscle quality of blunt snout bream after live transportation.

Facilitative glucose transporter 4 (GLUT4) plays a central role in mediating insulin function to increase glucose uptake in glucose metabolism homeostasis. In this study, the function and localization of GLUT4 in blunt snout bream (Megalobrama amblycephala) were first investigated, and then, the response measured as carbohydrate level, was analyzed. The results showed that the cDNA sequence of GLUT4 in blunt snout bream (MaGLUT4, GenBank accession no: MT447093) was 2868 bp in length, and the corresponding mRNA contained a 5′-UTR region of 513 bp and a 3′-UTR region of 837 bp. MaGLUT4 had an open reading frame of 1518 bp and was encoded by 505 amino acids. Its theoretical isoelectric point and molecular weight was 6.41 and 55.47 kDa, respectively. A comparison of these characteristics with BLASTP results from the NCBI database showed that MaGLUT4 had the highest homology with Cypriniformes fish, with MaGLUT4 and GLUT4 of other Cypriniformes clustered in the phylogenetic tree with other GLUT1–4 amino acid sequences. Compared with the results from the homo_sapiens and mus_musculus data sets, some mutations were observed in the GLUT4 amino acid sequence of these aquatic animals, including an FQQI mutation to FQQL, LL mutation to MM, and TELEY mutation to TELDY. MaGLUT4 was constitutively expressed in the muscle, intestine, and liver, with the highest mRNA level observed in muscle. Furthermore, the predicted tertiary structure and results of immunohistochemical staining showed that MaGLUT4 was a transmembrane protein primarily located in the plasma membrane, where it accounts for 60.9% of the total expressed, according to an analysis of subcellular localization. Blood glucose level peaked within 1 h, and the insulin level peaked at 6 h, while the mRNA and protein levels of GLUT4 showed an upward trend with an increase in feeding time and decreased sharply after 12 h. These results confirmed that MaGLUT4 was mainly distributed in muscles and crosses the cell membrane. The changes in the insulin, mRNA, and protein levels of MaGUT4 lagged far behind changes in blood glucose levels. This delay in insulin level changes and GLUT4 activation might be the important reasons for glucose intolerance of this fish species.

Cysteine cathepsins are described as lysosomal proteases with housekeeping and highly specialised functions in many organisms. In this study, cysteine cathepsins were identified from the transcriptome profile of blunt snout bream (Megalobrama amblycephala). A total of 41 cysteine cathepsin-like sequences were found and divided into groups of cathepsin (Cts): B, C, F, H, K, L, S, and Z. Twenty-nine of these sequences contained coding sequences, encoding 92-473 amino acids, which exhibited the highest (78%-95%) homology to the counterparts from zebrafish (Danio rerio). Multiple sequence alignment of the amino acids showed highly conserved domains among the cysteine cathepsins of M. amblycephala to its homologs. The putative proteins, including maCtsb1, maCtsc1, maCtsf2, maCtsh1, maCtsk5, maCtsl5, maCtss1, and maCtsz5 were characterised and structured using in silico methods. Additionally, codon usage of full length open reading frame sequences of these cathepsins was analysed. This study provided basic information for further studies on the specific functions of cysteine cathepsins of M. amblycephala in the future.

Abstract Background The blunt snout bream (Megalobrama amblycephala) is one of the most important commercial herbivorous fish in China, and dietary transition is an important event in blunt snout bream development. Gut microbiota has a vital role to host animal. However, little was known about the relationship among feeding habits transition, gut microbiota and digestive enzymes of gut content. Results In this study, 186,328 high-quality reads from nine 16S rRNA libraries were obtained using the Illumina MiSeq PE300 platform. The valid sequences were classified into 388 Operational Taxonomic Units, and a total of 223 genera, belonging to 20 phyla, were identified. The clustering result of gut bacterial communities is consistently related to the clustering result of intestinal content compositions. Proteobacteria and Firmicutes constitute the ‘core’ gut microbiota of blunt snout bream. Cetobacterium and Rhizobium were identified as microbiological markers of gut microbiota at zooplankton-based diet stages and diet transition stages, respectively. Moreover, thirteen potential cellulose-degrading bacteria were detected in our study. The canonical redundancy analysis (RDA) revealed that the feeding habits strongly influenced the gut microbiota and the digestive enzyme activities of gut content, while the result of PICRUSt test suggests that the metabolic capacity of gut microbiota was affected by feeding habit. Conclusions This study provided a comprehensive survey of the gut microbiota in blunt snout bream during its dietary transition period for the first time and clearly showed that the gut microbiota was strongly affected by feeding habit. This work allows us to better understand the relationship among gut microbiota, nutrition metabolism and feeding habits in vertebrate. Further, our study provides a reference for future studies investigating the metabolic adaption of herbivorous fish to shift to a vegetarian diet during their life history.

Copyright of Journal of Hydrobiology / Shuisheng Shengwu Xuebao is the property of Editorial Department of Journal of Hydrobiology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

An in-vitro study was carried out to examine the effects of yeast hydrolysate (YH) on antioxidant capacity and innate immunity of blunt snout bream (Megalobrama amblycephala) hepatocytes. Fish primary hepatocytes were seeded at a density of 3 × 105 cells mL−1 in 6-well tissue culture plates and treated with two different media including: 1) DMEM/F12 medium (control), and 2) YH medium [DMEM/F12 + 0.1 g L−1 YH]. After incubation for 24 h, the culture medium and primary hepatocytes were collected for subsequent analyses. The results showed no significant (P > 0.05) effect of YH on aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) activities and urea nitrogen (UN) concentration in the conditioned medium. However, significantly (P < 0.05) higher ALT and AST activities were found in YH treated hepatocytes compared to control. Moreover, YH supplementation led to significant enhancement of superoxide dismutase (SOD), catalase (CAT), alternative complement pathway (ACH50) and glutathione peroxidase (GPX) activities and reduction of malondialdehyde (MDA) concentration in the conditioned medium. Furthermore, YH application upregulated the expression of SOD , CAT and NOX2 genes and downregulated mRNA levels of Keap1 , Nrf2 and Bach1 in hepatocytes. Also, markedly higher lysozyme activity and albumin concentration were found in the conditioned medium of YH group compared to the control. Additionally, expression of immune-related genes such as antimicrobial peptides 1 (Leap 1) and Leap 2 were significantly upregulated by YH application. Down-regulated expression of NADPH oxidase-2 (NOX2), Kelch-like-ECH-associated protein 1 (Keap1), NF-E2-related factor 2 (Nrf2) and BTB and CNC homolog 1 (Bach1) were observed in YH treated hepatocytes. To conclude, YH supplementation improved antioxidant capacity and innate immunity of blunt snout bream hepatocytes. • YH supplementation improved hepatocytes metabolism. • YH boosted antioxidant activity of hepatocytes. • YH enhanced innate immunity of hepatocytes. [ABSTRACT FROM AUTHOR]

Facilitative glucose transporter 4 (GLUT4) plays a central role in mediating insulin function to increase glucose uptake in glucose metabolism homeostasis. In this study, the function and localization of GLUT4 in blunt snout bream (Megalobrama amblycephala) were first investigated, and then, the response measured as carbohydrate level, was analyzed. The results showed that the cDNA sequence of GLUT4 in blunt snout bream (MaGLUT4, GenBank accession no: MT447093) was 2868 bp in length, and the corresponding mRNA contained a 5′-UTR region of 513 bp and a 3′-UTR region of 837 bp. MaGLUT4 had an open reading frame of 1518 bp and was encoded by 505 amino acids. Its theoretical isoelectric point and molecular weight was 6.41 and 55.47 kDa, respectively. A comparison of these characteristics with BLASTP results from the NCBI database showed that MaGLUT4 had the highest homology with Cypriniformes fish, with MaGLUT4 and GLUT4 of other Cypriniformes clustered in the phylogenetic tree with other GLUT1–4 amino acid sequences. Compared with the results from the homo_sapiens and mus_musculus data sets, some mutations were observed in the GLUT4 amino acid sequence of these aquatic animals, including an FQQI mutation to FQQL, LL mutation to MM, and TELEY mutation to TELDY. MaGLUT4 was constitutively expressed in the muscle, intestine, and liver, with the highest mRNA level observed in muscle. Furthermore, the predicted tertiary structure and results of immunohistochemical staining showed that MaGLUT4 was a transmembrane protein primarily located in the plasma membrane, where it accounts for 60.9% of the total expressed, according to an analysis of subcellular localization. Blood glucose level peaked within 1 h, and the insulin level peaked at 6 h, while the mRNA and protein levels of GLUT4 showed an upward trend with an increase in feeding time and decreased sharply after 12 h. These results confirmed that MaGLUT4 was mainly distributed in muscles and crosses the cell membrane. The changes in the insulin, mRNA, and protein levels of MaGUT4 lagged far behind changes in blood glucose levels. This delay in insulin level changes and GLUT4 activation might be the important reasons for glucose intolerance of this fish species. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the effects of an extruded feed with or without anti-stress additives supplementation on the growth performance, digestive enzymes activities, stress response and the expressions of growth and feeding-related genes in blunt snout bream(Malobrama amblycephala).[Methods] Three kinds of feed with the same formula but different dosage forms were prepared by a feed manufacturing company following the practical production conditions, including a pelleted feed(PF),an extruded feed(EF) and the EF supplemented with an anti-stress additive(AF). A total of 15 000 fishes with an average body weight of 14 g were randomly distributed into three tanks(20 m×4 m×2 m). Then, fishes were randomly divided into three groups, including one offered the EF continuously, one fed the AF continuously and one fed the EF and PF during the first and last 90 days of the feeding trial(PE),respectively. Fishes were fed three times daily for 180 days. Samples were collected from intestines, blood and brain on the day 91 and day 181,respectively. Digestive enzyme activity, anti-stress ability and growth and feeding gene expression were measured.[Results] On the 91 day, the weight gain rate, body length and average daily gain of the AF group were all significantly higher than those of the other two groups(P<0.05),but the body composition showed no statistical difference among all the groups(P>0.05). The protease, lipase and amylase activities of the AF group were all significantly higher than those of the other two groups(P<0.05),but the plasma cortisol and lactic acid levels in the AF group were both significantly lower than those of the other two groups(P<0.05).On the 181 day, the weight gain rate and body length of the PE group were both significantly higher than those of the EF group(P<0.05),but the average daily gain of the PE group showed no statistical difference with that of the AF group(P>0.05). The condition factor and the viscera-body ratio of PE group were both significantly higher than those of the other two groups(P<0.05). The crude fat content of the PE group was significantly higher than those of the other two groups(P<0.05). The protease and lipase activities of the PE group were both significantly higher than those of the EF group(P<0.05),but showed no significant difference with those of the AF group(P>0.05). The plasma cortisol and lactic acid levels of EF group were both significantly higher than those of the other two groups(P<0.05),and showed no statistical difference between the other two groups. In addition, no significant difference was observed in the mRNA expression of leptin in liver among all the three groups(P>0.05),but the mRNA expressions of insulin-like growth factor in liver and growth hormone receptor and neuropeptide Y in pituitary in the EF group were all significantly lower than those in the other two groups(P<0.05). The mRNA expression of growth hormone in the PE group was significantly higher than that in the other groups(P<0.05),while the mRNA expression of cholecystokinin in the EF group was significantly higher than that in the other two groups(P<0.05).[Conclusions] Compared with the EF group, feeding the AF feed could improve the growth performance and anti-stress ability of blunt snout bream. Besides, compared with the EF group, the growth performance and growth hormone gene expression of blunt snout bream could also be enhanced by feeding the extruded feed and pelleted feedat equal time intervals. [ABSTRACT FROM AUTHOR]

The study investigated whether dietary berberine supplementation could improve intestinal barrier against inflammation induced by high-fat and high-carbohydrate diets in blunt snout bream. Fish (average initial weight 44.83 ± 0.06 g) were fed with six kinds of diets (control, high-fat diet (10% lipid) and high-carbohydrate (43% nitrogen-free extract) diet, control/berberine, high-fat/berberine or high-carbohydrate/berberine) for 8 weeks, respectively. Feeding mode of berberine (50 mg/kg diet) was adopted to two-week interval. After feeding trial, fish growth performance and intestinal barrier function were estimated. The result showed that no significant interactions between diet and berberine in growth performance, whole body composition or protein utilization were observed (P > 0.05). Specific growth rate (SGR) and feed conversion ratio (FCR) were significantly affected by berberine (P < 0.05). Protein efficiency ratio (PER), nitrogen retention (NRE), fish whole-body lipid contents increased greatly in high-fat or high-carbohydrate diets (P < 0.05). Significant interactions between diet and berberine were observed in fish intestinal barrier (physical, chemical, immunological and microbiological barriers) (P < 0.05). High-fat and high-carbohydrate diets could increase significantly intestinal permeability and inflammatory response, decrease intestinal mucins gene expression levels, and make the intestinal microbiota out of balance (P < 0.05). Berberine significantly inhibited inflammation response and modulated intestinal microflora profile (P < 0.05). Taken together, berberine could alleviate intestinal barrier damage injured by high-fat or high-carbohydrate diet and improve the growth performance of blunt snout bream. • Dietary berberine supplementation at two-week interval feeding mode could protect intestinal barrier damage induced by high-fat diet and high-carbohydrate diet. [ABSTRACT FROM AUTHOR]

Four diets were formulated with or without fishmeal (FM) and essential amino acid (EAA) supplementation fed to the blunt snout bream throughout 8 weeks. The results showed that total cholesterol level (TC) was decreased significantly against FM reduction (P < 0.05). Aspartate aminotransferase (AST), alanine transaminases (ALT), and alkaline phosphatase (ALP) activities were affected neither by FM reduction nor EAA supplementation (P > 0.05). Catalase (CAT) activity and glutathione (GSH) content were significantly augmented and diminished in the group fed diet 3 respectively (P < 0.05). Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content did not alter either by FM reduction or EAA supplementation (P > 0.05). Glutathione peroxidase (GPx) activity was elevated significantly either by reducing FM or supplementing EAA (P < 0.05). The relative expression of nuclear factor erythroid 2-related factor 2 (Nrf2), Kelch-like-ECH-associated protein 1 (Keap1), and GPx was significantly augmented (P < 0.05) in the EAA-supplemented groups. Neither FM reduction nor EAA supplementation altered the relative expression of SOD (P > 0.05), whereas the relative expression of copper-zinc superoxide dismutase (Cu/Zn-SOD) decreased against FM reduction (P < 0.05). The relative expressions of Interleukin 8 (IL-8), tumour necrosis factor-α (TNF-α), interleukin 10 (IL10), and nuclear factor-kappa B (NF-κB) were diminished significantly in FM-reduced groups (P < 0.05). The supplementation of EAA significantly increased the relative expression of signal transducer and activator of transcription 4 (STAT4) (P < 0.05). In conclusion, the results revealed that dietary plant proteins with EAA supplementation significantly affected the physiological status and immune responses in blunt snout bream. [ABSTRACT FROM AUTHOR]

In terms of taxonomic status, common carp (Cyprinus carpio, Cyprininae) and crucian carp (Carassius auratus, Cyprininae) are different species; however, in this study, a newborn homodiploid crucian carp-like fish (2n=100) (2nNCRC) lineage (F1−F3) was established from the interspecific hybridization of female common carp (2n=100)×male blunt snout bream (Megalobrama amblycephala, Cultrinae, 2n=48). The phenotypes and genotypes of 2nNCRC differed from those of its parents but were closely related to those of the existing diploid crucian carp. We further sequenced the whole mitochondrial (mt) genomes of the 2nNCRC lineage from F1 to F3. The paternal mtDNA fragments were stably embedded in the mt-genomes of F1−F3 generations of 2nNCRC to form chimeric DNA fragments. Along with this chimeric process, numerous base sites of F1−F3 generations of 2nNCRC underwent mutations. Most of these mutation sites were consistent with the existing diploid crucian carp. Moreover, the mtDNA organization and nucleotide composition of 2nNCRC were more similar to those of the existing diploid crucian carp than those of the parents. The inheritable chimeric DNA fragments and mutant loci in the mt-genomes of different generations of 2nNCRC provided important evidence of the mtDNA change process in the newborn lineage derived from hybridization of different species. Our findings demonstrated for the first time that the paternal mtDNA were transmitted into the mt-genomes of homodiploid lineage, which provided new insights into the existence of paternal mtDNA in the mtDNA inheritance. [ABSTRACT FROM AUTHOR]

Toll-like receptors (TLRs) are a category of pattern recognition receptors (PRRs), which recognize pathogen associated molecular patterns (PAMPs) and participate in the immune responses. We identified tlr5a , tlr5b , tlr9 and tlr21 from the genome of blunt snout bream (Megalobrama amblycephala). All four tlrs were constitutively expressed in all examined tissues. After an immune bacterial challenge with Aeromonas hydrophila , their expressionwas up-regulated in lymphoid organs and tissues. Recombinant eukaryotic plasmid pEGFP-N1 was transfected into the common carp (Cyprinus carpio) EPC (epithelioma papulosum cyprini) cells for the purpose of subcellular localization. pcDNA3.1(+) recombinant eukaryotic plasmid was used to investigate the effects of overexpression of tlrs on the expression of downstream interferon-associated immune factors. The four Tlrs were distributed in the cytoplasm of transfected cells and appeared as filamentous or reticular. The expression of irf3 , irf7 , isg15 , mx1 , pkr and viperin at 0, 6, 12, 18, 24, 36, 48 and 72 h post-transfection in transfected EPC cells was quantified by qPCR. Overexpression of tlrs upregulated the expression of viperin , isg15 , irf3 , irf7 , mx1 and pkr (in that order of magnitude). We also cloned the following promoters of irfs : Irf1-p , irf2-p , irf6-p , irf7-p , irf8-p and irf9-p. Results of the dual luciferase reporter assay suggested that tlr5a , tlr5b and tlr9 enhanced the activities of irf7-p , while tlr5b enhanced the activities of irf1-p and irf7-p. This suggests that they all play a role in the innate immunity. The experiments also indicated that TLRs activate irf3 or irf7 signaling to induce IFN secretion and subsequent upregulation of IFN-stimulated genes. These results indicate that tlrs and irfs play an important immune role in response to A. hydrophila infection in blunt snout bream, and pave the way for further studies of immune mechanisms mediated by TLRs in fish. • tlr5a, tlr5b, tlr9 and tlr21 cDNA were cloned from Megalobrama amblycephala. • tlrs from M. amblycephala are constitutively expressed in a wide range of tissues in vivo. • Aeromonas hydrophila infection alters the mRNA expression of tlrs. • Overexpression of tlr genes upregulated the expression of irf3 , irf7 , isg15 , mx1 , pkr and viperin. • Cloned six promoters of irfs in M. amblycephala and tlr5a , tlr5b and tlr9 enhanced the activities of irf7-p. [ABSTRACT FROM AUTHOR]

Fish meal is the main protein source in fish diet, but due to its unsustainability, it has to be replaced with more sustainable protein sources, such as soybean meal (SBM), in fish feeds. As SBM often produces negative effects on fish growth and health, we conducted an eight-week trial of complete fish meal replacement with SBM in the diet of juvenile (48.2 ± 0.55 g) blunt snout bream (Megalobrama amblycephala), a herbivorous cyprinid fish native to the Yangtze basin (China). SBM-fed fish exhibited: a worsened growth performance; improved survival rate (P > 0.05); elevated serum triglycerides and total cholesterol but reduced low-density lipoprotein in plasma (P < 0.05); mild lipid droplet accumulation and swelling of the liver; elevated goblet cell counts and mild signs of enteritis in the intestines. With negligible effects on growth and mild effects on health, M. amblycephala appears to be comparatively tolerant of SBM, but total fish meal replacement is not recommended. Future studies should aim to identify the SBM tolerance threshold in long-term growth trials. [ABSTRACT FROM AUTHOR]

Juvenile blunt snout bream Megalobrama amblycephala were reared under three photoperiods, 8 L: 16D, 12 L: 12D and 16 L: 8D (L: light; D: dark) for over 8 weeks. The results showed that growth performance was significantly enhanced by long photoperiod. Contrary to feed conversion ratio, final length, final weight, weight gain and feed intake increased significantly as illumination time increased from 8 h daily to 16 h daily. Low size heterogeneity and whole-body lipid content in fish exposed to long photoperiod were also observed. Both relative mRNA expression level of GH and IGF-I increased as illumination time increased from 8 h to 16 h daily. The lowest value of plasma cortisol was observed at the middle the photophase while the highest value was observed at the transition between the day and night span. Hepatic MDA content significantly increased as illumination time increased from 8 h daily to 16 h daily. The activities of hepatic catalase and glutathione peroxidase were lowest in fish exposed to 16 L: 8D photoperiod and significantly lower than that in fish exposed to 8 L: 16D. These results indicate that photoperiod manipulation may not only improve growth performance but also reduce size heterogeneity. However, prolonged photoperiod could cause chronic stress since plasma cortisol level was higher in the long photoperiod group, leading to an increasing oxidative stress. • Long photoperiod not only improved growth performance but also reduced size heterogeneity. • Long photoperiod caused the elevated mRNA expression of GH and IGFI-I. • Long photoperiod may cause chronic stress, leading to oxidative stress. [ABSTRACT FROM AUTHOR]

The CITED3 protein is a non-DNA-binding transcriptional co-regulator involved in the regulation of various transcriptional responses against hypoxia stress. Here, we characterized two paralogs Cited3 genes (Cited3a and Cited3b) from blunt snout bream (Megalobrama amblycephala), which is a hypoxia-sensitive species. Both genes have an open reading frame of 756 and 723 bp; encoded a protein of 251 amino acid and 240 amino acid, respectively; and they shared a sequence identity of 67%. In adult fish, both Cited3a and Cited3b mRNAs were highly expressed in kidney tissues. In contrast, they were detected in the skin, muscle, and gonad at extraordinarily low levels. During embryogenesis, both Cited3a and Cited3b mRNAs were maternally deposited in eggs and fluctuated from the zygote to the 44-hpf (hours post-fertilization) larvae. Whole-mount in situ hybridization demonstrated that both Cited3a and Cited3b mRNAs were transcribed in the brain, gut, and tailbud at 12 hpf, and at the brain and gut at 24 hpf, and at the brain at 36 hpf embryos. Hypoxic treatment led to upregulated expression of the Cited3 genes during embryogenesis. Under hypoxia, both Cited3a and Cited3b genes in the kidney and brain and Cited3a genes in the liver were significantly upregulated. These results suggest that hypoxia was associated with increases in mRNA levels for both Cited3a (kidney, brain, liver) and Cited3b (kidney and liver). [ABSTRACT FROM AUTHOR]

This study aimed to investigate the effects of Streptococcus faecalis on the growth performance, intestinal histology, antioxidant capability, innate immunity, and disease resistance of blunt snout bream Megalobrama amblycephala. Fish were fed five experimental diets containing 0 (SF0, control), 1 × 105 (SF1), 1 × 106 (SF2), 1 × 107 (SF3), and 1 × 108 cfu/g (SF4) of Streptococcus faecalis, respectively, for 10 weeks. After the feeding trial, fish were challenged by Aeromonas hydrophila with the cumulative mortality recorded during a period of 96 h. The results showed that WG and FI of fish both showed no significant difference (P > 0.05) among all the treatments. However, the FCR was significantly (P < 0.05) affected by Streptococcus faecalis levels with the lowest value observed in the SF2 group, whereas the opposite was true for intestinal microvillus length (P < 0.05). Dietary supplementation of 1 × 106 cfu/g Streptococcus faecalis significantly (P < 0.05) increased the hepatic activities of SOD, CAT, and GPx; plasma activities of LZM, MPO, ACP, and AKP; and the levels of C3, C4, and IgM of fish, compared with the control group. Similar results were also observed in the tissue expressions of Leap-I, Leap-II, muc2, and muc5b (P < 0.05), whereas the opposite was true for liver MDA contents and plasma NO levels (P < 0.05). At 96 h after challenge, the cumulative mortality of the control was significantly (P < 0.05) higher than that of the SF2 group, but it showed no statistical difference (P > 0.05) with that of the other treatments. These results indicated that dietary supplementation of 1 × 106 cfu/g Streptococcus faecalis could not only improve the feed utilization of blunt snout bream but also enhance its antioxidant capability, innate immunity, and disease resistance. [ABSTRACT FROM AUTHOR]

The present study evaluated the mechanisms governing insulin signaling, glucose metabolism, and lipogenesis in juvenile fish fed with different dietary leucine levels. Fish were fed six practical diets with graded leucine levels ranging from 0.90 to 2.94% of dry basis for 8 weeks. The trial results showed that, compared to the control group (0.90%), optimal dietary leucine level (1.72%) resulted in the up-regulation of mRNA expression related to insulin signaling pathway, including target of rapamycin (TOR), insulin receptor substrate 1 (IRS-1), phosphoinositide 3-kinase (PI3K), and protein kinase B (Akt). However, an excessive leucine level (2.94%) led to protein S6 kinase 1 (S6K1) overexpression and inhibited TOR, IRS-1, PI3K, and Akt mRNA expressions. The protein level of TOR, S6K1, IRS-1, PI3K, and Akt showed a similar result with mRNA level of these genes. Optimal dietary leucine level (1.72%) significantly improved plasma insulin content, while high level of leucine showed an inhibiting phenomenon. Optimal dietary leucine level (1.72%) could reduce plasma glucose by enhancing the ability of glycometabolism including improving glucose transporter 2 (GLUT2), glucokinase (GK) expressions and down-regulating phosphoenolpyruvate carboxykinase (PEPCK) expression. While an excessive leucine level (2.94%) resulted in high plasma glucose by inhibiting the ability of glycometabolism including lowering GLUT2 and GK expressions, and improving glucose-6-phosphatase (G6Pase) and PEPCK expressions. The relative expressions of pyruvate kinase (PK) and glycogen synthase (GS) were not significantly affected by dietary leucine levels. Dietary leucine level of 1.33% could improve plasma triglyceride content (TG) by enhancing lipogenesis including improving sterol-response element-binding protein 1 (SREBP1), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), and glucose-6-phosphate dehydrogenase (G6PDH) expressions compared to the control group (0.90%). Total cholesterol (TC) was not significantly affected by dietary leucine levels. The present results indicate that optimal leucine level could improve glycolysis and fatty acid synthesis through improving insulin sensitivity in juvenile blunt snout bream. However, excessive dietary leucine level resulted in high plasma glucose, which led to insulin resistance by inhibiting the gene expressions of insulin signaling pathway and activating gluconeogenesis-related gene expression. [ABSTRACT FROM AUTHOR]

Abstract Interferon regulatory factor (Irf) family represents one of the most important transcription factor families, with multiple biological roles. In this study, we characterized five Irf family members (irf4a, irf4b, irf6, irf8 and irf10) in Megalobrama amblycephala at the cDNA and (predicted) amino acid levels, analyzed them phylogenetically, and developed gene-specific primers for qPCR analysis. All five irfs were constitutively expressed in all examined tissues, but their transcription was significantly higher in lymphoid organs and tissues, such as kidney, spleen and intestine. Exceptions were irf8 , which was expressed at a high level in heart and brain tissues, and irf6 , expressed at low levels in most tissues. After a bacterial immune challenge with Aeromonas hydrophila, the expression of irfs in liver was up-regulated: mairf4a 8.12-fold , mairf4b 29.9-fold , mairf6 1.38-fold and mairf10 1.65-fold (mairf8 was an exception: 0.07-fold). In spleen, kidney, intestine and gills, transcript levels of studied irfs increased only at specific time-points. The results suggested that irfs are involved in the immune response to bacterial infection in M. amblycephala , which will help elucidate the biological functions of irfs in the immune system of teleost fish. Highlights • Irf4a , irf4b, irf6, irf8 and irf10 cDNA were cloned from Megalobrama amblycephala. • Mairfs genes and proteins share significant structural homology with the known homologs. • Mairfs are constitutively expressed in a wide range of tissues in vivo. • Aeromonas hydrophila infection alters the mRNA expression of mairfs. [ABSTRACT FROM AUTHOR]

Abstract This study was conducted to understand the effect of high-fat diet challenge on lipid transport and endoplasmic reticulum stress in blunt snout bream. Ninety fish (average weight: 41.84 ± 0.07 g) were randomly fed a control diet (6% fat) or a high-fat diet (11% fat) for 9 weeks. The growth performance and feed utilization efficiency were evaluated at the end of the trial. The liver samples of both groups were harvested for molecular analysis and histological evaluation. Compared to the Control group, the high-fat diet group showed no effects on either growth performance or energy intake in blunt snout bream. However, high-fat diet resulted in a massive accumulation of lipid and pathological structural alternations, and disrupted expression of lipid transport-related genes and endoplasmic reticulum stress in the liver of the fish. In vitro, after exposure of the isolated primary hepatocytes from blunt snout bream to oleic acid, the cells showed increased intracellular TG accumulation, decreased VLDL secretion, which was attributed to altered expression levels of lipid transport-related genes through the activated IRE1/XBP1 signaling. The oleic acid-induced detrimental effects were alleviated by co-incubating the cells with an IER1 inhibitor, 4μ8c. In conclusion, high-fat diet could lead to aberrant lipid secretion by activating the ER stress-associated IRE1/XBP1 pathway. Inhibiting the activity of IRE1 represents a promising target to rescue the side-effects of high-fat diet on the liver function of blunt snout bream. Highlights • High-fat diets could result in abnormal secretion and synthesis of VLDL protein. • High-fat diet could lead to aberrant lipid secretion by activating the ER stress-associated IRE1/XBP1 pathway. • Inhibiting the activity of IRE1 represents a promising target to rescue the side-effects of high-fat diet on fish. [ABSTRACT FROM AUTHOR]

Abstract RNase1 is an enzyme important in host defense in vertebrates where it degrades the RNA of bacteria and viruses. We evaluated the effect of RNase1 on the resistance to Aeromonas hydrophila infection in Megalobrama amblycephala. The fish were randomly divided into four groups: a blank group (none-treated M. amblycephala), a control group (injected PBS), a challenge group (A. hydrophila- injected) and a treatment group (pre-treated with RNase1 24 h before the A. hydrophila injection), and we collected five tissues of each group. Then we recorded changes in the levels of glutathione (GSH), oxidized glutathione (GSSG), hepatic catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA) and lysozyme; and the relative mRNA expression of catalase (CAT), selenium-dependent glutathione peroxidase (GPx), Cu/Superoxide dismutase (Cu/Zn-SOD), glutamate-cysteine ligase (GCLC), glutathione reductase (GR) and nuclear factor erythroid 2-related factor 2 (Nrf2) for four groups. The expression of six genes was highest in liver and blood of the blank group. It was significantly higher in the gut of the treatment group (compared to control and challenge groups) 12 h after the infection. The treatment group exhibited a significant increase in GSH, SOD and CAT activity, and a decrease in GSSG, MDA and lysozyme content (compared to the control and challenge groups) 6 and 12 h after infection. These results suggest that supplementation with RNase1 protein can enhance resistance against A. hydrophila infections in M. amblycephala. Highlights • CAT, GPx, Cu/Zn-SOD, GCLC, Nrf2 and GR genes in fish and mammals were analyzed. • These genes were highly expressed in liver and blood of healthy M.amblycephala. • Pre-treatment with RNase1 increase GSH, CAT and SOD activity after A.hydrophila infection. • Pre-treatment with RNase1 decrease GSSG, MDA and lysozyme content after A.hydrophila infection. • RNase1 protein play a protective role in M. amblycephala after A. hydrophila infection. [ABSTRACT FROM AUTHOR]

Six diets were formulated to investigate the success of fish meal (FM) replacement by plant proteins; diet 1 reflected a commercial feed (8% FM), diet 3 contained 4% FM, and diet 5 was devoid of FM. Whereas, diet 2, diet 4, and diet 6 reflected diet 1, diet 3, diet 5, respectively, and supplemented with essential amino acid (EAA). At the end of 8‐week trial, there was no significant difference in survival rate. Significantly higher final weight, weight gain rate, and specific growth rate were recorded in the group fed diet 2 compared with the other treatments (except diet 4) (p < 0.05). Feed conversion ratio of fish fed diet 2 was significantly lower than those fed diets 1, 3, and 5 (p < 0.05). The lowest feed intake and highest protein efficiency rate were found in fish fed diet 2 (p < 0.05). There was no significant difference in whole body compositions between treatments. Plasma aspartate transaminases, alanine aminotransferase, and glucose were significantly affected by dietary treatments (p < 0.05), while plasma protein and albumin contents were not influenced by the treatments. The relative expression of target of rapamycin (TOR) and phosphatidylinositol 3‐kinase, regulatory subunit 1 (alpha) (PIK3R1) in fish fed diet 3 (4% FM) were significantly down‐regulated compared with those fed diet 6 for TOR and diets 4 and 6 for PIK3R1 (p < 0.05). Insulin receptor substrate 1 (IRS‐1) and janus kinase 3 (JAK3) expressions were fluctuated, with the higher levels in fish fed diets 4 and 6. In conclusion, the findings of this study indicate that plant protein mixture supplemented with EAA could be used to substitute FM in practical diet for Megalobrama amblycephala. [ABSTRACT FROM AUTHOR]

PGC1α is a transcriptional coactivator that plays key roles in mitochondrial biogenesis, so exploring its molecular characterization contributes to the understanding of mitochondrial function in cultured fish. In the present study, a full-length cDNA coding PGC1α was cloned from the liver of blunt snout bream (Megalobrama amblycephala) which covered 3741 bp with an open reading frame of 2646 bp encoding 881 amino acids. Sequence alignment and phylogenetic analysis revealed high conservation with other fish species, as well as other higher vertebrates. Comparison of the derived amino acid sequences indicates that, as with other fish, there is a proline at position 176 (RIRP) compared to a Thr in the mammalian sequences (RIRT). To investigate PGC1α function, three in vitro tests were carried out using primary hepatocytes of blunt snout bream. The effect of AMPK activity on the expression of PGC1α was determined by the culture of the hepatocytes with an activator (Metformin) or inhibitor (Compound C) of AMPK. Neither AMPK activation nor inhibition altered PGC1α expression. Knockdown of PGC1α expression in hepatocytes using small interfering RNA (si-RNA) was used to determine the role of PGC1α in mitochondrial biogenesis. No significant differences in the expression of NRF1 and TFAM, and mtDNA copy number were found between control and si-RNA groups. Also, hepatocytes were cultured with oleic acid, and the findings showed the significant reduction of mtDNA copy number in oleic acid group compared to control. Moreover, oleic acid down-regulated the expression of NRF1 and TFAM genes, while PGC1α expression remained unchanged. Our findings support the proposal that PGC1α may not play a role in mitochondrial biogenesis in blunt snout bream hepatocytes.

This study investigated the effects of resveratrol on the growth performance, energy sensing, glycolipid metabolism and glucose and insulin load of blunt snout bream Megalobrama amblycephala fed high-carbohydrate diets. Fish (39.44 ± 0.06 g) were randomly fed three diets: a control diet (30% carbohydrate), a high-carbohydrate diet (HC, 41% carbohydrate), and the HC diet supplemented with 0.04% resveratrol (HCR) for 12 weeks. Fish fed the HC diet had significantly high values of nitrogen and energy retention efficiency, hepatosomatic index, intraperitoneal fat ratio, whole-body lipid content and intraperitoneal fat glycogen and lipid contents compared to the control group, but showed little difference with the HCR treatment. Liver and muscle lipid contents and plasma levels of glucose, glycated serum protein, advanced glycation end products and total cholesterol of fish fed the HC diet were significantly higher than those of the control group, whereas the opposite was found with resveratrol supplementation. Fish fed the HC diet obtained significantly low values of plasma insulin levels and hepatic adenosine monophosphate (AMP) contents and NAD+/NADH ratio compared to HCR treatment, but showed little difference with the control group. The opposite was found for hepatic adenosine triphosphate (ATP) contents and the ATP/AMP ratio. In addition, fish fed the HC diet showed significantly high transcriptions of glucose transporter 2 (GLUT2), glucose-6-phosphate dehydrogenase, glycogen synthase, fatty acid synthetase (FAS), acetyl-CoA carboxylase α (ACCα), peroxisome proliferator-activated receptor γ and PPARα compared to the control group, whereas the opposite was found for protein levels of AMP-activated protein kinase α (t-AMPKα), phosphorylated AMP-activated protein kinase α (p-AMPKα), sirtuin-1 (SIRT1), and p-AMPKα/t-AMPKα ratio as well as the transcriptions of AMPKα1, AMPKα2, SIRT1, PPARγ coactivator-1α (PGC-1α), phosphoenolpyruvate carboxykinase, fructose-1,6-bisphosphatase (FBPase), glucose-6-phosphatase, carnitine palmitoyl transferase I (CPT I) and acyl-CoA oxidase. Resveratrol supplementation significantly up-regulated the protein levels of t-AMPK, p-AMPK, and SIRT1, p-AMPK/t-AMPK ratio as well as the transcriptions of AMPKα1, AMPKα2, SIRT1, PGC-1α, GLUT2, FBPase, and CPT I compared to HC group, while the opposite was found for sterol regulatory element-binding protein-1, FAS and ACCα. Furthermore, resveratrol improved glucose and insulin tolerance of fish fed the HC diet after glucose and insulin load.

The proteins CITED belong to a family of non-DNA-binding transcriptional co-regulators involved in the regulation of various transcriptional responses. Previous studies suggest that members of CITED family may function in response to hypoxia in mammals. however, the molecular and functional information on CITED genes in aquaculture fish is unclear. Here, we characterized and examined the transcriptional patterns of CITED1 and CITED2 genes in the hypoxia-sensitive blunt snout bream (Megalobrama amblycephala). Blunt snout bream CITED1 and CITED2 genes shared a relatively low sequence identity of 45%. CITED1 and CITED2 mRNAs were widely transcribed in adult tissues. During embryogenesis, CITED1 mRNA was significantly transcribed at 4, 24, 28, 40, and 44 hpf, whereas CITED2 mRNA levels fluctuated from the zygote to 44 hpf larval stage. Whole-mount in situ hybridization demonstrated that CITED1 and CITED2 mRNAs were detected in the brain at 12 hpf, brain and gut at 24 hpf, and brain at 36 hpf. In addition, low expression of CITED1 mRNA was detected in the tailbud at 24 hpf. The results of acute hypoxia experiment showed that CITED1 and CITED2 mRNAs were markedly upregulated in the kidney and downregulated in the liver, brain, gill, and heart under hypoxia. Embryos in hypoxic conditions at different developmental stages showed a significant increase in mRNA levels of CITED1 and CITED2. These results provide a new insight into the divergence of CITED1 and CITED2 genes and their transcriptional responses to hypoxia.

Abstract Currently, there is a trend to use high-fat diets in intensive aquaculture that is accompanied with incidence of fatty liver when dietary lipid level surpasses an upper limit. So, it is necessary to develop appropriate strategies to reduce the risk of fatty liver in commercial fish farming. Studies in mammals have revealed a correlation between fatty liver and AMP-activated protein kinase (AMPK) activity, which has been recognized as a key modulator of lipid metabolism. Considering the frequent occurrence of fatty liver in blunt snout bream farming, an in-vitro study was designed to evaluate the efficiency of metformin, as a stimulator of AMPK, in activation of AMPK and its subsequent effects on lipid metabolism in primary hepatocytes. Fish hepatocytes were seeded at a density of 1 × 106 ml−1 in 6-well tissue culture plates and treated with three different media including: 1) Leibovitz's L-15 medium [L15] as control, 2) high-fat medium [L15 + 400 μM oleic acid], and 3) metformin medium [L15 + 400 μM oleic acid+200 μM metformin]. After 48 h of culture, the cells and supernatant were collected for analysis. The results showed significant (P <.05) enhancement of cell triglyceride and total cholesterol concentrations in the high-fat group over control, and metformin addition significantly reduced the values. Also, the high-fat group exhibited significantly higher aspartate aminotransferase activity than both control and metformin groups. The lowest AMPK and phospho-AMPK protein expression was found in the high-fat group while metformin addition significantly up-regulated their expression levels. Mitochondrial and peroxisomal oxidation rates in the high-fat group were significantly lower than control while similar oxidation rates were observed for metformin treated and control groups. The high-fat group showed significantly lower CPT I activity than control, and metformin inclusion increased the activity. Expression of genes associated with lipid metabolism such as PPARα, CPT I, AOX, PGC-1α and TFAM was suppressed in the high-fat group, and metformin supplementation up-regulated their expression levels. The opposite trend was true for the expression of ACC2 gene. Also, the results showed down-regulation of FAS and SREBP-1C genes in the high-fat medium group, and metformin addition resulted in further reduction of their expression level. The lowest activities of mitochondrial complexes (I-III) were found in the high-fat group and metformin prevented high-fat-induced reduction of mitochondrial complexes activity. Notably increased concentrations of reactive oxygen species and malondialdehyde were found in the high-fat group, and metformin treatment reduced their concentrations. Moreover, metformin group exhibited higher glutathione peroxidase activity than the high-fat group. The findings in this study showed clearly that metformin activated AMPK in blunt snout bream hepatocytes, which contributed to enhanced lipid metabolism and attenuated lipid deposition in the cells incubated with high-fat medium. Highlights • Metformin was tested for AMP-activated protein kinase (AMPK) activation in fish. • Metformin reduced triglyceride and cholesterol concentrations in hepatocytes. • AMPK and phospho-AMPK protein expression was significantly enhanced by metformin. • Metformin increased fatty acid oxidation and lipid metabolism genes expression. [ABSTRACT FROM AUTHOR]

Abstract p38 mitogen-activated protein kinase (MAPK) is an important protein which plays a key role in regulating the innate immunity, so exploring its molecular characterization is helpful in understanding the resistance against microbial infections in cultured fish. Here, a full-length cDNA of p38 MAPK was cloned from liver of blunt snout bream (Megalobrama amblycephala) which covered 2419 bp with an open reading frame of 1086 bp encoding 361 amino acids. p38 MAPK contained the characteristic structures of Thr-Gly-Tyr (TGY) motif and substrate binding site Ala-Thr-Arg-Trp (ATRW), which are conserved in MAPK family. To investigate p38 MAPK functions, two in vivo experiments were carried out to examine its expression following ammonia exposure and bacterial challenge. Also, an in vitro experiment was conducted to assess the role of p38 MAPK in inflammation of primary hepatocytes induced by lipopolysaccharide (LPS). The results showed the ubiquitous expression of p38 MAPK in all the tested tissues with varying levels. p38 MAPK mRNA expression was significantly up-regulated by ammonia stress and Aeromonas hydrophila challenge, and altered in a time-dependent manner. Moreover, the results indicated that the inflammatory response induced by LPS in hepatocytes is p38 MAPK dependent as knockdown of p38 MAPK using siRNA technology depressed the expression of IL-1β and IL-6. The findings in this study showed that p38 MAPK has anti-stress property, and plays key role in protection against bacterial infection and inflammation in blunt snout bream. Highlights • p38 MAPK was characterized in blunt snout bream. • p38 MAPK expression was explored after ammonia stress and bacterial challenge. • p38 MAPK role was tested in inflammation of primary hepatocytes. • p38 MAPK was up-regulated by ammonia stress and A. hydrophila challenge. • Knockdown of p38 MAPK using siRNA lowered the inflammatory response. [ABSTRACT FROM AUTHOR]

Abstract This study aimed to investigate the effects of dietary non-protein energy adjustments on the mitochondrial biosynthesis and function of juvenile Megalobrama amblycephala. Fish (average weight: 37.98 ± 0.07 g) were fed eight diets containing two dietary carbohydrate levels (30% and 43%) and four lipid sources (fish oil, soybean oil, palm oil and the mixed oil) for 11 weeks. Liver mitochondrial respiratory chain complex V activity and ATP (adenosine triphosphate) content both increased significantly with increasing dietary carbohydrate levels, whereas the opposite was true for the AMP (adenosine 5′-monophosphate)/ATP ratio, hepatic transcripts of AMP-activated protein kinase α1 (AMPKα1), AMPKα2, peroxisome proliferators γ-activated receptor coativator-1α (PGC-1α), NADH dehydrogenase 1 and cytochrome c oxidase 1 (COX1) as well as the activities of Na+-K+-ATPase, succinate dehydrogenase (SDH), citrate synthase (CS) and mitochondrial respiratory chain complex I, III and IV. Additionally, hepatic ATP content, the transcripts of AMPKα, COX1 and ATP6 and the activities of Na+-K+-ATPase, SDH, CS and mitochondrial respiratory chain complex III were all significantly affected by lipid sources. Furthermore, an interaction between dietary carbohydrate levels and lipid sources was also observed in the activities of liver mitochondrial Na+-K+-ATPase and respiratory chain complex III as well as the transcripts of ATP6 and PGC-1α. Overall, these findings suggested that dietary carbohydrate levels and lipid sources remarkably affected the mitochondrial biosynthesis and function of M. amblycephala. A diet containing 30% carbohydrate and FO could boost its mitochondrial biosynthesis, while that of 30% carbohydrate and SO could enhance the mitochondrial function. [ABSTRACT FROM AUTHOR]

This study investigated the effects of resveratrol on the growth performance, energy sensing, glycolipid metabolism and glucose and insulin load of blunt snout bream Megalobrama amblycephala fed high-carbohydrate diets. Fish (39.44 ± 0.06 g) were randomly fed three diets: a control diet (30% carbohydrate), a high-carbohydrate diet (HC, 41% carbohydrate), and the HC diet supplemented with 0.04% resveratrol (HCR) for 12 weeks. Fish fed the HC diet had significantly high values of nitrogen and energy retention efficiency, hepatosomatic index, intraperitoneal fat ratio, whole-body lipid content and intraperitoneal fat glycogen and lipid contents compared to the control group, but showed little difference with the HCR treatment. Liver and muscle lipid contents and plasma levels of glucose, glycated serum protein, advanced glycation end products and total cholesterol of fish fed the HC diet were significantly higher than those of the control group, whereas the opposite was found with resveratrol supplementation. Fish fed the HC diet obtained significantly low values of plasma insulin levels and hepatic adenosine monophosphate (AMP) contents and NAD+/NADH ratio compared to HCR treatment, but showed little difference with the control group. The opposite was found for hepatic adenosine triphosphate (ATP) contents and the ATP/AMP ratio. In addition, fish fed the HC diet showed significantly high transcriptions of glucose transporter 2 (GLUT2), glucose-6-phosphate dehydrogenase, glycogen synthase, fatty acid synthetase (FAS), acetyl-CoA carboxylase α (ACCα), peroxisome proliferator-activated receptor γ and PPARα compared to the control group, whereas the opposite was found for protein levels of AMP-activated protein kinase α (t-AMPKα), phosphorylated AMP-activated protein kinase α (p-AMPKα), sirtuin-1 (SIRT1), and p-AMPKα/t-AMPKα ratio as well as the transcriptions of AMPKα1, AMPKα2, SIRT1, PPARγ coactivator-1α (PGC-1α), phosphoenolpyruvate carboxykinase, fructose-1,6-bisphosphatase (FBPase), glucose-6-phosphatase, carnitine palmitoyl transferase I (CPT I) and acyl-CoA oxidase. Resveratrol supplementation significantly up-regulated the protein levels of t-AMPK, p-AMPK, and SIRT1, p-AMPK/t-AMPK ratio as well as the transcriptions of AMPKα1, AMPKα2, SIRT1, PGC-1α, GLUT2, FBPase, and CPT I compared to HC group, while the opposite was found for sterol regulatory element-binding protein-1, FAS and ACCα. Furthermore, resveratrol improved glucose and insulin tolerance of fish fed the HC diet after glucose and insulin load. [ABSTRACT FROM AUTHOR]

An 11-week feeding trial was carried out to investigate the effects of supplemented chromium picolinate (Cr-Pic) on the growth, whole-body composition, and relative mRNA expression related to lipogenesis and glucose metabolism in juvenile blunt snout bream. Seven isonitrogenous and isoenergetic diets with graded Cr supplementation levels were fed to triplicate groups. The final weight (FW), feed conversion ratio (FCR), and specific growth rate (SGR) were improved with increasing dietary Cr supplementation levels up to 0.4 mg/kg, and thereafter showed relatively constant. However, 12.0 mg/kg dietary Cr supplementation decreased growth and feed utilization. Based on SGR and FCR, the optimal dietary Cr supplementation level for the juvenile was estimated to be 0.28 mg/kg. Significantly higher plasma insulin levels were found in juvenile fed diets with 0.4 and 0.8 mg/kg Cr supplementation compared to those fed diet sans supplemented Cr. Plasma glucose levels decreased with increasing dietary Cr supplementation, and the lowest value was remarked in the group added 3.2 mg/kg of Cr. Adding 0.4-0.8 mg/kg Cr enhanced insulin receptor substrate 1 (IRS-1), phosphoinositide-3-kinase (PI3K), and pyruvate kinase (PK) and inhibited expression of phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G6Pase), and glycogen synthase (GS) mRNA levels. High dietary Cr (12.0 mg/kg) supplementation resulted in high G6Pase and PEPCK expression. The highest content of whole-body lipid was remarked in fish fed with 0.4 mg/kg dietary Cr, which related to the enhanced gene expression related to lipogenesis; thereafter, mRNA levels showed a diminishing trend. These findings indicate that optimum dietary Cr-Pic supplementation has a positive effect on growth and blood glucose homeostasis by modifying the mRNA levels related to glucose metabolism and lipogenesis in juvenile blunt snout bream. [ABSTRACT FROM AUTHOR]

The aims of the present study were to clone the full-length cDNA of adipose triglyceridelipase (ATGL) and to analyze its expression after lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-α). The cDNA obtained covered 1801 bp with an open reading frame of 1500 bp encoding 499 amino acids. Sequence alignment and phylogenetic analysis show the best identity with Cyprinus carpio (86%). The ATGL protein shared a highly conserved 169-amino acid patatin domain, containing a glycine-rich motif, an active serine hydrolase motif, and an aspartic active site. The highest ATGL expression was observed in the liver followed by muscle, whereas relatively low values were detected in the brain and adipose. TNF-α is regarded as an important factor in regulating fat metabolism. Here, LPS was used to induce TNF-α in vivo to verify whether TNF-α can affect ATGL expression. TNF-α expression in liver and muscle is increased and remains unchanged in adipose tissue and brain. The variation of ATGL activity is consistent with that of TNF-α gene expression. Next, we explored the mechanism by which LPS-induced TNF-α mediates the mRNA expression of ATGL in the liver and muscle. For liver, the mRNA levels of c-Jun N-terminal kinase (JNK), nuclear factor kappa B (NF-κB), Sirtuin 1 (SIRT1), and AMP-activated protein kinase (AMPK) were increased by LPS-induced TNF-α. Differencing from the situation in the liver, there was a near-significant decrease trend in the expression of SIRT1 in muscle. Those results indicated that the ATGL gene of blunt snout bream shared a high similarity with the other vertebrates. The expression level of ATGL in tissues with high-fat content was intended to be high. LPS can induce ATGL expression perhaps related to TNF-α. [ABSTRACT FROM AUTHOR]

Abstract This study aimed to investigate the effects of fishmeal (FM) replacement by rice protein concentrate (RPC) with a xylooligosaccharides supplement on the growth performance, antioxidant capability and immune response against Aeromonas hydrophila in blunt snout bream ( Megalobrama amblycephala ). Fish (46.85 ± 0.34 g) were randomly assigned to one of 6 diets, namely the control diet (containing FM), the RPC diet (FM replaced by RPC) and RPC diet supplemented with 0.5, 1.5, 2.3 and 3% XOS respectively, for 8 weeks. After the feeding trial, fish were challenged by Aeromonas hydrophila for 96 h with the blood and liver sample obtained at 48 and 96 h respectively. The results showed that the final weight, weight gain and protein efficiency ratio of fish fed RPC diet were significantly ( P < 0.05) lower than that of the control group, whereas the opposite was true for FCR. However, the supplement of 1.5% XOS remarkably ( P < 0.05) improved these parameters compared to the control diet. Plasma total iron binding capacity of fish fed the RPC diet showed little difference ( P > 0.05) with that of the control group, but it enhanced significantly ( P < 0.05) with the supplement of 1.5% XOS compared to the control group. After bacterial infection, plasma lysozyme (LYM), complement 3, complement 4, myeloperoxidase (MPO), acid phosphatase (ACP) and alkaline phosphatase activities, as well as immunoglobulin M, levels all increased significantly ( P < 0.05) with the maximum value is attained at 48 h, then they decreased significantly ( P < 0.05) with further increasing time at 96 h. Similar results were also observed in liver superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) activities as well as malondialdehyde (MDA) content. Regarding dietary treatment, these parameters of fish fed RPC diet showed little difference ( P > 0.05) with those fed the control diet but were significantly enhanced ( P < 0.05) when RPC diet was supplemented with 1.5–2.3% XOS, except for hepatic (MDA) content which showed an opposite trend compared to the control group. After 96 h of challenge, the relative percentage survival (RPS) of fish fed XOS was significantly higher ( P < 0.05) than that of fish offered the control and RPC diet. In addition, significant ( P < 0.05) interactions between dietary XOS and sampling time were also observed in plasma LYM, MPO, ACP, and hepatic SOD, CAT, GPX, MDA, as well as RPS. In conclusion, this study indicated that dietary supplementation of 1.5%XOS could significantly improve the growth performance, antioxidant capability, innate immunity and A. hydrophila resistance of blunt snout bream fed diets with FM replaced by RPC. Highlights • Optimum feeding of xylooligosaccharides (XOS) enhanced growth performance in blunt snout bream. • The best immunity and antioxidant status was observed in fish fed rice protein concentrate supplemented with 1.5–2.3% XOS. • Excess XOS tend to cause immune suppression as well as reduced A. hydrophila resistance. • Relative percentage survival rate of fish fed XOS was significantly higher than control diets. [ABSTRACT FROM AUTHOR]

Abstract: An 8‐wk feeding trial was conducted to evaluate the influence of different proportions of dietary plant ingredients on the growth of juvenile blunt snout bream, Megalobrama amblycephala. A 2 × 3 factorial arrangement was set for this trial with three ratios of cottonseed meal (CSM) and canola meal (CM) (2:1, 1:1, or 1:2) and three proportions of soybean meal (SBM) (2, 1, or 0). The results showed that the specific growth rate and protein efficiency ratio both significantly decreased with decreasing amounts of SBM in the diets of blunt snout bream (P < 0.05), while the feed efficiency ratio and feed intake increased significantly (P < 0.05). Meanwhile, the growth performance and feed utilization were shown to improve with increasing amounts of CSM, although the effects were not significant (P > 0.05). The interaction of these two factors had no effect on either growth or feed utilization (P > 0.05). Diet composition was not seen to affect body composition (P > 0.05). The ratio of CSM to CM significantly affected gut amylase activity, liver alanine aminotransferase content, and plasma cholesterol levels (P < 0.05). The interaction between the two main effects significantly influenced the plasma glucose level (P < 0.05). Therefore, the suitable proportion of plant feed for blunt snout bream should include high levels of SBM and CSM, with a SBM : CSM : CM ratio of 2:2:1. [ABSTRACT FROM AUTHOR]

Zinc (Zn) is widely known as an essential trace element for fish and new ways to supply it to them are needed. Palygorskite (Pal) is a natural silicate clay mineral and the palygorskite structure contains nano-channels, which are filled with water and exchangeable ions. Zn-bearing palygorskites (Zn-Pal) prepared using ion exchange have attracted attention due to the durable antibacterial properties that limit pathogens and as a potential new Zn source for livestock. The present study was conducted to evaluate the effects of Zn-Pal supplementation on the growth performance, nutrient retention, meat quality, Zn accumulation, and intestinal Zn transporter protein gene expression in blunt snout bream Megalobrama amblycephala. The fish were fed a basal diet without an exogenous Zn source and the basal diet was supplemented with 125 mg/kg Zn as Zn sulfate (ZnSO4) or 35, 80, or 125 mg/kg Zn as Zn-Pal. Each diet was tested using three replicates for 7 weeks. The results showed that dietary Zn-Pal supplementation quadratically (P<0.05) increased growth performance, nutrient retention, total and Cu/Zn superoxide dismutase activity, Zn content in scales, and intestinal Zn transporter protein gene expression. The muscular cooking loss in blunt snout bream decreased with the optimum Zn-Pal Zn level of 35 mg/kg. Compared to the fish treated with ZnSO4, the fish supplemented with 35 mg/kg as Zn-Pal exhibited similar growth performance and nutrient retention (P>0.05), increased mRNA expression of the metal-response element-binding transcription factor-1 in the intestine (P<0.05), and decreased cooking loss of muscle (P<0.05).The results suggested that 35 mg/kg Zn supplementation as Zn-Pal could improve the growth performance and body composition, increase nutrient retention and tissue Zn concentrations, enhance the muscle water-holding capacity, and enhance antioxidant status in blunt snout bream. The Zn-Pal was more efficient and could be used as an alternative Zn source to ZnSO4 in the diet of blunt snout bream. KeyWords-Blunt Snout Bream, Growth Performance, Meat Quality, Zinc-bearing Palygorskite. [ABSTRACT FROM AUTHOR]