Xi Zhang

Diet form is increasingly recognized as a welfare-relevant factor in intensive aquaculture, yet the effects of feed cooking on fish behavioral and physiological welfare remain poorly characterized. Juvenile southern catfish (Silurus meridionalis; 6.18 ± 0.52 g) were reared for 6 weeks in an indoor recirculating aquaculture system and fed either raw grass carp (Ctenopharyngodon idella) muscle (fish fed raw muscle, FR) or cooked grass carp muscle (fish fed cooked muscle, FC; 15 min ramp to ~100 °C followed by 2–3 min at ~100 °C). Locomotor activity and anxiety-like behavior were assessed using the open-field test and an annular light–dark preference assay, respectively. Flow-through respirometry further revealed a significantly lower standard metabolic rate (SMR) in FC fish than in FR fish, decreasing from 10.30 to 6.83, which represents a 33.7% reduction. Endocrine and biochemical analyses showed that cooking significantly decreased serum total triiodothyronine (T3) by 23.8%, whereas routine serum biochemical indices remained unchanged. In brain tissue, dopamine (DA) was significantly reduced by 7.2% in the FC group, and RT-qPCR analysis of dopamine-related genes further showed a significant downregulation of the rate-limiting synthesis gene th. These results indicate that cooking primarily downshifts the activity-energy axis in southern catfish and is accompanied by coordinated thyroid and dopaminergic changes. To our knowledge, this is the first integrated study to evaluate the behavioral, metabolic, and neuroendocrine effects of cooked feed in S. meridionalis, providing a short-term phenotypic baseline for assessing welfare-relevant feeding scenarios in aquaculture.

The gel-forming ability of collagens is vital for their application in cell scaffolds, yet very few comparative studies on fish collagen sources are available. This study isolated and characterized type I collagens from carp skin (CSK), scales (CSC), and swim bladders (CSB) and sturgeon skin (SSK) and swim bladders (SSB). The carp collagens exhibited higher thermal stability (34.75–34.78 °C) and formed more transparent, stronger gels than the sturgeon collagens. Additionally, as demonstrated by scanning electron microscopy, the sturgeon collagens exhibited faster fibril formation, with visible fibrils after 3 h which grew thicker but did not form bundles. The carp collagens, in contrast, initially displayed fewer, thinner, and longer fibrils, with their formation accelerating over time and fibril bundles emerging after 24 h. All collagen solutions of 4% (w/v) exhibited shear-thinning flow behavior, with the carp-derived solutions showing higher viscosities (10³–10⁴ Pa·s) than those demonstrated by the sturgeon-derived solutions (10²–10³ Pa·s). The CSBs and SSBs demonstrated the highest storage (G′) and loss (G″) moduli, with the former exhibiting the lowest loss tangent (tan δ), indicative of a stronger gel structure. The gels at 24 h showed slightly poorer mechanical properties than those at 3 h. The CSC and SSB gels had the highest thermal stability. These findings highlight the distinctiveness of the characteristics of collagens and their gels, emphasizing their potential in biomaterial applications. The present study also provides a foundational framework for assessing cellular responses in a comparative context that may help in identifying the most suitable collagen types for biomedical applications.

To investigate the effects of photoperiods on the growth and muscle quality indicators of grass carp (Ctenopharyngodon idella), 225 fish (109.65 ± 3.62 g) were randomly assigned into five different photoperiod groups (0L:24D, 8L:16D, 12L:12D, 16L:8D, and 24L:0D). The experiment spanned a 75-day period, after which sampling and analysis were performed. Compared with the 0L:24D and 8L:16D groups, the 12L:12D and 16L:8D groups significantly promoted the growth of grass carp (p < 0.05). The texture parameters of the muscle in the 0L:24D and 16L:8D groups were significantly greater than those in the 12L:12D group (p < 0.05). The crude protein content was significantly higher in the 12L:12D and 16L:8D groups (p < 0.05). The amino acid content and muscle fiber characteristics, as well as the mRNA levels of myostatin (mstn), myogenic factor 5 (myf5), type I collagen α1 (col1α1), and α2 (col1α2), along with the hydroxyproline and collagen contents, were all significantly influenced by the photoperiod (p < 0.05). The lysine (Lys), aspartic acid (Asp), and alanine (Ala) contents in the muscle and muscle fiber density of grass carp reached the highest levels under the 16L:8D treatment (p < 0.05). Collectively, these results indicate that a 16L:8D photoperiod is optimal for enhancing both the growth and muscle quality indicators of grass carp. The findings of this study offer valuable scientific references for the precise regulation of grass carp quality when using a photoperiod, and they are anticipated to foster the further development and optimization of strategies for improving grass carp quality.

The southern catfish (Silurus meridionalis) is an economically important carnivorous freshwater fish in China. In this study, we compared the properties of skin collagen from southern catfish fed with raw food (RF) and cooked food (CF). The skin collagen yield in the RF group (8.66 ± 0.11%) was significantly higher than that of the CF group (8.00 ± 0.27%). SDS-PAGE, circular dichroism spectroscopy, and FTIR analyses revealed that the collagen extracted from southern catfish skin in both groups was type I collagen, with a unique triple helix structure and high purity. The thermal denaturation temperature of collagen in the RF group (35.20 ± 0.11 °C) was significantly higher than that of the CF group (34.51 ± 0.25 °C). The DPPH free radical scavenging rates were 68.30 ± 2.41% in the RF collagen and 61.78 ± 3.91% in the CF collagen, which was higher than that found in most fish collagen. Both the RF and CF groups had high ability to form fibrils in vitro. Under the same conditions, the CF group exhibited faster fibril formation and a thicker fibril diameter (p < 0.05). In addition, the RF group exhibited significantly higher expression of col1a1 compared to the CF group. These results indicated that feeding southern catfish raw food contributed to collagen production, and the collagen from these fish may have potential in biomaterial applications.

The biochemical properties of collagens and gels from grass carp (Ctenopharyngodon idella) were studied to explore the feasibility of their application in biomaterials. The yields of skin collagen (SC) and swim bladder collagen (SBC) extracted from grass carp were 10.41 ± 0.67% and 6.11 ± 0.12% on a wet basis, respectively. Both collagens were characterized as type I collagen. Denaturation temperatures of SC and SBC were 37.41 ± 0.02 °C and 39.82 ± 0.06 °C, respectively. SC and SBC had high fibril formation ability in vitro, and higher values of salinity (NaCl, 0–280 mM) and pH (6–8) in formation solution were found to result in faster self-assembly of SC and SBC fibrils as well as thicker fibrils. Further tests of SC gels with regular morphology revealed that their texture properties and water content were affected by pH and NaCl concentration. The hardness, springiness, and cohesiveness of SC gels increased and the chewiness and water content decreased as pH increased from 7 to 8 and NaCl concentration increased from 140 to 280 mM. These properties suggest that collagens from grass carp may be useful in biomaterial applications in the future.

Ammonia is one of the most important environmental factors in aquatic ecosystems. However, there are limited studies on the effects of chronic or long-term ammonia stress and its potential molecular mechanism in fish. This study aimed to investigate the immune response and molecular mechanisms in the spleen and head-kidney of fish following chronic ammonia exposure. Megalobrama amblycephala (9.98 ± 0.48 g) were exposed to different concentrations of total ammonia nitrogen (0–30 mg/L) for 30 days. Ammonia exposure caused significant increases in cortisol levels and decreases in lysozyme and complement 3/4 concentrations in the serum, indicating inhibitory effects of ammonia stress on innate immune responses. Ammonia exposure also induced concentration-dependent increases in ammonia concentrations in tissue, pathological damage and indexes of spleen and head-kidney. Additionally, the contents of immunoglobulin M (IgM), interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) as well as mRNA levels of toll-like receptors (TLRs)/Myeloid differentiation factor 88 (MyD88)-independent signaling molecules in the spleen and head-kidney were significantly downregulated after ammonia exposure. Our findings suggested that chronic ammonia exposure caused the suppression of innate and adaptive immune responses through downregulating TLR/MyD88-independent signaling. Adverse influences of chronic ammonia stress were more severe in the spleen than in the head-kidney.

Heat shock protein 47 (Hsp47) is a collagen-specific molecular chaperone that is indispensable for molecular maturation of collagen. In this study, hsp47 and hsp47-like cDNAs were cloned and characterized in grass carp (Ctenopharyngodon idella). The cDNAs were 1212 and 1218 base pairs long, respectively, and included an open reading frame encoding 403 and 405 amino acids. The molecular phylogeny based on the deduced amino acid sequences indicated that the correct sequences of the hsp47 and hsp47-like cDNA were obtained and the deduced proteins clustered distinctly into teleost clades. Primary structure analysis and characterization of Hsp47 and Hsp47-like shared the basic structure and biofunctions of Hsp47 in vertebrates. The spatial pattern of gene expression revealed that hsp47 and hsp47-like were relatively ubiquitous in different tissues and highly expressed in heart and skin. The expression levels of hsp47 and hsp47-like and type I collagen mRNAs varied similarly in different tissues. Type I collagen content increased significantly with the increase of water velocity in the muscle of grass carp in response to aerobic exercise. Among the gene expression changes of hsp47, hsp47-like, col1a1 and col1a2 in muscle that occurred in response to aerobic exercise, the change of type I collagen was most strongly correlated with hsp47 expression. Additionally, col1a1 showed the highest correlation with hsp47-like and col1a2 showed the highest correlation with hsp47. These findings suggest that grass carp Hsp47 and Hsp47-like are closely related to type I collagen synthesis. This study firstly suggests fish aerobic exercise can improve type I collagen content and Hsp47 gene expression in muscle of grass carp.

Fish muscle quality is an important parameter in the aquaculture industry. In this study, we analyzed and compared the muscle quality of grass carp (Ctenopharyngodon idellus) cultured at salinities of 0‰, 3‰, and 6‰ (GC0, GC3, GC6). There was no significant difference in crude protein and crude fat content of muscle between GC0 and GC3. Crude fat was significantly lower in GC6 compared to the other groups. GC3 and GC6 had higher hydroxyproline content, which suggested that these groups had higher collagen content. GC3 and GC6 had higher contents of free amino acids and umami amino acids than GC0, but there was no significant difference in sweet or sour amino-acid content among groups. GC3 and GC6 had better texture properties, including hardness, gumminess, chewiness, resilience, and springiness, than GC0. GC3 had the highest water-holding capacity among the groups. As the salinity increased, the diameter of muscle fibers decreased and the sarcolemma showed a thickening trend. These results suggest that a slight increase in salinity (i.e., 3‰) can effectively improve the muscle quality of grass carp.