Molecular Characterization of Prdx Family and Response of Antioxidant Enzymes in Berberine Hydrochloride-Treated Charybdis japonica Infected With Aeromonas hydrophila

Berberine hydrochloride is an isoquinoline alkaloid, which has antitumoral, antibacterial, and antiviral activities in vivo and in vitro. Charybdis japonica is one of the main economic species of crab in Southeast Asia. We studied the molecular mechanism of oxidative stress in berberine hydrochloride-treated C. japonica infected with Aeromonas hydrophila. C. japonica were infected with A. hydrophila after being submerged in different concentrations (0, 100, 200, and 300 mg/L) of berberine hydrochloride for 48 h. The full-length cDNA of Prx6 and the ORFs of Prx5 and PXL2A were cloned. Prx6 and PXL2A each have one conserved domain, Cys44, and Cys81. The Prx5 conserved domain contains three important Cys loci, Cys75, Cys100, and Cys76. Prx6 was different from Prx5 and PXL2A in the Peroxiredoxin family. The transcription levels of PXL2A infected with A. hydrophila were all higher than the control. The transcription levels of C. japonica were further increased by adding berberine hydrochloride and were increased the highest at a concentration of 300 mg/L. The activities of glutathione peroxidase, superoxide dismutase, and catalase in the hepatopancreas of berberine hydrochloride-treated C. japonica infected with A. hydrophila were significantly increased compared with those only infected with A. hydrophila and the control group. The glutathione transferase activity in the hepatopancreas was significantly increased in berberine hydrochloride-treated C. japonica. The results of this study provide a new understanding of the potential role of berberine hydrochloride on the oxidative stress mechanisms of C. japonica.

Effect of Berberine Hydrochloride Treatment on Transcriptomic Response in Gill Tissue of Charybdis japonica Infected With Aeromonas hydrophila

Berberine hydrochloride is the main active ingredient of Coptis chinensis, which has demonstrated significant antibacterial activity against bacteria. However, the molecular mechanism underlying its effects is unclear in crabs. In this study, we used transcriptome sequencing to investigate the molecules involved in the therapeutic and defensive responses of Charybdis japonica infected with 105 colony-forming units (CFU)/L of Aeromonas hydrophila following treatment with berberine hydrochloride. A total of 5,409 differentially expressed genes were observed after exposure of C. japonicus for 72 h to 100 mg/L berberine hydrochloride, 100 mg/L berberine hydrochloride plus injection of 105 CFU/L A. hydrophila, and a control group injected with equal amount of physiological saline solution. Enrichment analysis revealed that these genes were involved in metabolism, cellular processes, signal transduction, and immune function. The transcriptomic results indicated that exposure to berberine hydrochloride activated glutathione metabolism, oxidative phosphorylation, fatty acid degradation, retinol metabolism, pyruvate metabolism, and the hypoxia-inducible factor 1 signaling pathway. Transcriptomic analysis and relative mRNA level analysis also identified acid hydrolase genes and phagosomal genes as key factors regulating the adaptation of acid berberine-infused C. japonicus to infection with A. hydrophila, Based on the ability of berberine hydrochloride to induce acid hydrolase and phagosomal gene expression, which can in turn remove bacteria, immersion in berberine hydrochloride may remove A. hydrophila and thus improve the survival of C. japonica. The results of this study provided a new scientific basis for the potential role of berberine hydrochloride in the immune mechanisms of crabs.

Effect of Aeromonas hydrophila infected on metabolomic response and gut microbial 16S rRna of Charybdis japonica

The innate immune response of Charybdis japonica treated with Aeromonas hydrophila was explored using bioinformatics. Metabolomics data were integrated with a gut microbial 16S rRNA dataset, together with information on corresponding enzyme activity. The results of the study showed that after being infected with A. hydrophila, some beneficial genera of bacteria in the gut of C. japonica, such as Photobacterium, Rhodobacter, Polaribacter, Psychrilyobacter, Mesoflavibacter, Fusibacter and Phormidium, could directly inhibit Vibrio or produce extracellular polysaccharides with highly effective antibacterial properties. The intestinal probiotics of C. japonica such as Mesoflavibacter have a mutually reinforcing relationship with Phaeobacter, Colwellia, Bacillus, Psychrobacter and Cohaesibacter. Conditional pathogenic bacteria in the gut of healthy crabs may also have such a symbiotic relationship with intestinal probiotics, promoting their growth and reproduction. For example, Phormidium has a mutualistic relationship with Aeromonas and Azopira. Metabolites in the gut of C. japonica infected with A. hydrophila, including beta-alanine metabolism, nitrogen metabolism, inositol phosphate metabolism, galactose metabolism, histidine metabolism, ascorbate and arginine and proline metabolism were increased, with alanine metabolism being the most abundant. The activity of metabolite related enzymes such as lipid peroxidase, phenoloxidase, superoxide dismutase, nitric oxide synthase, glutathione transferase and mid-glutathione decreased and NO levels also decreased. The positive correlation with the probiotic flora suggests that metabolites increase with bacterial abundance and that microbial metabolites or co-metabolites can, in turn, achieve many pleiotropic effects to resist invasion by A. hydrophila. These results may contribute to further research in the resistance of C. japonica to invading pathogens.

Improving size selectivity of round pot for Charybdis japonica by configuring escape vents in the Yellow Sea, China

Sustainable development of the important economic species, Asian paddle crab (Charybdis japonica), has attracted attention in the coastal waters of the Yellow Sea, China. The commonly used round pots are almost nonselective, resulting in severe bycatch of juveniles. In this study, we explored a method to improve the size selectivity for C. japonica by mounting escape vents on the side panels of each pot. The selectivity of pots with escape vent sizes of 70 mm × 20 mm, 70 mm × 25 mm, 70 mm × 30 mm, and 70 mm × 35 mm was tested using a catch comparison method. The estimated minimum landing size (MLS) of carapace height (27 mm), according to the regulated MLS of carapace length (50 mm), was used as a reference point to explain the results. Significant increases in the size of crabs caught by pots were found with the enlargement of escape vent size (Kruskal–Wallis test, P < 0.01). The pots with 70 mm × 20 mm, 70 mm × 25 mm and 70 mm × 30 mm escape vents released nearly 50%, 75% and 95% of undersized individuals, respectively, and these three types of pots retained approximately 90% of legal-sized individuals compared with the control pots without escape vents. The pots with 70 mm × 35 mm escape vents released nearly all undersized individuals, but they also released most legal-sized individuals. Pots with an escape vent size of 70 mm × 30 mm were recommended for the sustainable development of C. japonica in the Yellow Sea of China. The results of this study reiterate the importance of carapace height for determining the size selectivity, which can serve as a reference to formulate management regulations in the coastal waters of the Yellow Sea, China.