In aquaculture, the use of probiotics to improve growth, immunity, and stress resistance in crustaceans has gained increasing attention. This study examined the effects of incorporating different levels of Rhodotorula mucilaginosa (0.0, 0.1, 1.0, and 10.0 g/kg) into the diet on growth performance, antioxidant capacity (AOC), immune function, Toll/Imd, and JAK‐STAT signaling pathways in red claw crayfish ( Cherax quadricanatus ). The investigation was conducted through a 56‐day feeding trial. The main results are as follows: Compared with the control group (0.0 g/kg), different R. mucilaginosa levels significantly increased ( p < 0.05) the specific growth rate (SGR) and weight gain rate (WGR) of red claw crayfish, significantly increased ( p < 0.05) the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione S‐transferase (GST), total AOC (T‐AOC), and acid phosphatase (ACP) in hepatopancreas of red claw crayfish, and significantly upregulated ( p < 0.05) the relative expression levels of tumor necrosis factor receptor‐associated protein 6, akirin, immunodeficiency homolog, interferon regulatory factor 4, Toll‐like receptor (TLR) 6, TLR 2, Janus kinase, signal transducer activator of transcription, tumor necrosis factor α, interleukin‐1β, transforming growth factor‐β1 genes in the hepatopancreas of red claw crayfish. In conclusion, R. mucilaginosa significantly enhanced red claw crayfish’s growth, AOC, and immune function, and activated the Toll/Imd and JAK‐STAT signaling pathways. In this experimental context, the ideal addition level of R. mucilaginosa is 1.0 g/kg.

Mannan oligosaccharide (MOS), a prebiotic derived from yeast cell walls, has been shown to enhance growth performance and health status in various aquatic species. As an exogenous antigen adjuvant, MOS modulates T-cell-mediated immune responses, thereby improving immune function and suppressing excessive inflammatory reactions. This study aimed to evaluate the effects of dietary MOS supplementation on growth performance, serum biochemical parameters, muscle composition, digestive enzyme activity, antioxidant and immune status, and the mTOR signaling pathway in juvenile GIFT tilapia (Oreochromis niloticus). Juveniles (initial body weight: 16.17 ± 1.32 g) were randomly assigned to six treatment groups (three replicate tanks per group) and fed diets supplemented with MOS at 0, 0.2%, 0.4%, 0.6%, 0.8%, and 1% (equivalent to 0, 2, 4, 6, 8, and 10 g/kg of diet, respectively) for 60 days. Compared with the control group, fish fed MOS-supplemented diets exhibited significantly higher (p < 0.05) weight gain rates, specific growth rates, and protein efficiency ratios, along with a significantly lower (p < 0.05) feed conversion ratio. Serum albumin, high-density lipoprotein, and lysozyme levels were significantly increased (p < 0.05), whereas triglycerides, low-density lipoprotein, aspartate aminotransferase, and alanine aminotransferase levels were significantly decreased (p < 0.05). In the liver, head kidney, and spleen, the expression of pro-inflammatory genes (tumor necrosis factor α, interleukin 1β, interleukin 6, interleukin 8, and interferon γ) was significantly downregulated (p < 0.05), while the expression of antioxidant and protective genes (superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, nuclear factor erythroid 2-related factor 2, lysozyme, alkaline phosphatase, interleukin-10, transforming growth factor β, and heat shock protein 70) as well as mTOR signaling pathway-related genes (mammalian target of rapamycin, akt protein kinase B, phosphatidylinositol 3 kinase, and ribosomal protein S6 kinase polypeptide 1) was significantly upregulated (p < 0.05). Overall, MOS positively affects tilapia’s growth, health, and immunity, with 0.60% identified as the optimal dietary level based on growth performance.

The growth environment significantly influences the intestinal microbiota of aquatic organisms. We investigated the composition and functional differences in the intestinal microbiota of red claw crayfish (Cherax quadricarinatus) in rice fields (RB) and ponds (PB) by 16S rDNA high-throughput sequencing technology. The results indicate that the Shannon, Simpson, Sobs, Chao1, and ACE indices of PB are all higher than those of RB, demonstrating greater diversity and richness of intestinal microbiota. The dominant phyla in the intestinal microbiota of the Cherax quadricarinatus were Proteobacteria, Tenericutes, and Firmicutes. Tenericutes and Proteobacteria were significantly more abundant in the RB than in the PB, while Planctomycetes and Firmicutes were significantly more abundant in the PB than in the RB. The results of network correlation analysis indicate that Proteobacteria and Firmicutes exhibit strong connectivity with other microbial groups in the gut microbiota of Cherax quadricarinatus, showing significant centrality. They play an important role in the interactions within the gut microbiota community. The dominant bacterial genera in the Cherax quadricarinatus’s gut were Citrobacter, Candidatus_Bacilloplasma, and Clostridium_sensu_stricto_1. The abundance of the genus Clostridium was significantly higher in the PB than in the RB, whereas the abundance of Candidatus_Hepatoplasma and Vibrio was significantly lower in the PB than in the RB. The prediction function of KEGG enrichment showed that the abundance of Amino acid metabolism, Biosynthesis of Other Secondary Metabolites, Transport and Catabolism, Cancers, and Nervous System, Substance Dependence were significantly higher in the PB, while the infectious diseases pathway was enriched in the RB. In summary, our results revealed significant differences in the composition and diversity of intestinal microbiota in the Cherax quadricarinatus between rice paddy and pond farming environments. The intestinal microbiota of the Cherax quadricarinatus grown in pond environments exhibit higher diversity and stability, manifested by an increase in beneficial bacteria abundance and a decrease in opportunistic pathogens. These findings significantly improve understanding of the complex relationship among Cherax quadricarinatus, intestinal microbiota, and the environment.

This study investigated the effects of dietary supplementation with varying levels (CK: 0.0 g/kg; RL: 0.1 g/kg; RM: 1.0 g/kg; RH: 10.0 g/kg) of Rhodotorula mucilaginosa on muscle composition, serum biochemical indicators, antioxidant capacity, lipid metabolism, and the mTOR signaling pathway in red claw crayfish (Cherax quadricarinatus). Results showed that, compared to CK, treatment groups had higher muscle crude protein, fat, leucine, histidine, arginine, and essential amino acids (p < 0.05), and lower saturated fatty acids (p < 0.05). Treatment groups also exhibited increased activities of alkaline phosphatase, acid phosphatase, superoxide dismutase, catalase, glutathione S-transferase, lysozyme, albumin, total protein, and antioxidant capacity (p < 0.05), with reduced activities of aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and triglycerides (p < 0.05). In the hepatopancreas, treatment groups showed significant downregulation of AMP-activated protein kinase α, β, and γ, and carnitine palmitoyltransferase 1 genes (p < 0.05). Conversely, genes involved in lipid anabolism (peroxisome proliferator-activated receptor γ, acetyl-CoA carboxylase, fatty acid synthase, sterol regulatory element-binding protein, protein kinase B, and mammalian target of rapamycin 1 and 2) were upregulated (p < 0.05). In conclusion, R. mucilaginosa supplementation affects muscle composition, lipid metabolism, and mTOR signaling. The optimal dose is 1.0 g/kg.

The objective of this study was to evaluate the effects of dietary supplementation with different levels of Rhodotorula mucilaginosa (0.0 g/kg, 0.1 g/kg, 1.0 g/kg, and 10.0 g/kg) on resistance to Aeromonas veronii infection in red claw crayfish (Cherax quadricarinatus) (initial body weight of 0.13 ± 0.06 g). The investigation combined a 56-day feeding trial with a subsequent 7-day infection challenge to assess cumulative mortality, immune and antioxidant enzyme activities, and the relative expression of immune-related genes. During the A. veronii infection test, the cumulative mortalities for the 0.1 g/kg, 1.0 g/kg, and 10.0 g/kg groups were 44.44%, 38.89%, and 38.89%, respectively, all significantly lower (p < 0.05) than that of the control group (58.33%). Compared with the control group, after infection with A. veronii, the activities of acid phosphatase, alkaline phosphatase, catalase, and superoxide dismutase in the hepatopancreas and alkaline phosphatase, lysozyme in the hemolymph of red claw crayfish in the 1.0 g/kg group significantly increased (p < 0.05). The activities of aspartate aminotransferase and alanine aminotransferase in the hemolymph of red claw crayfish in the 1.0 g/kg group significantly decreased (p < 0.05). The relative expression levels of serine protease inhibitor, crustacean hyperglycemic hormone, anti-lipopolysaccharide factor, and superoxide dismutase genes in the hepatopancreas of red claw crayfish in the 1.0 g/kg group were significantly upregulated (p < 0.05). In conclusion, R. mucilaginosa could significantly improve the antibacterial ability of red claw crayfish against A. veronii. In this experimental context, the ideal addition level of R. mucilaginosa is determined to be 1.0 g/kg.

Gonadotropin releasing hormone (GnRH) plays an important role in the regulation of vertebrate reproduction. Studies have shown that immunization against GnRHa can induce sexually sterile tilapia. To explore the mechanism behind this, in this study, RNA-seq and data-independent acquisition (DIA) techniques were used to study the transcriptome and proteome of the gonad of tilapia immunized with GnRHa. 644 differentially expressed genes (80 upregulated and 564 downregulated) and 1150 differentially expressed proteins (351 upregulated and 799 downregulated) were identified. There were 209 genes with consistent differential expression patterns in the transcriptomic and proteomic analyses, of which 9 were upregulated and 200 downregulated, indicating that the gonad gene expression was inhibited by GnRHa immunization. The downregulated genes were particularly involved in the functions of single-organism process, binding, cellular process, metabolic process and catalytic activity, and associated with the pathways including ECM-receptor interaction, focal adhesion, cardiac muscle contraction and oxidative phosphorylation. The expression of six differentially expressed genes involved in the GnRH signaling pathway was all downregulated. In addition, several important functional genes related to gonadal development after GnRHa immunization were screened. This study confirmed the expression of corresponding genes was affected by GnRHa on the gonad development in tilapia at the molecular level, and laid a foundation for elucidating the mechanism of GnRHa immunization.