In order to understand the effects of fermented Astragalus membranaceus (FAM) on the liver and intestinal health of tiger grouper (Epinephelus fuscoguttatus), this study was conducted. This study evaluates the effects of different levels of FAM on liver and intestinal tissue structure, serum biochemical parameters, intestinal digestive enzyme, and microbiota structure of tiger grouper. Fish were fed with diets (crude protein ≥ 48.0%, crude fat ≥ 10.0%) with five levels of FAM (L1:0.25%, L2: 0.5%, L3: 1%, L4: 2% and L5: 4%) in the experimental groups and a regular diet was used as the control (L0: 0%) for 8 weeks. Compared with AM, the protein content of FAM was significantly changed by 34.70%, indicating that a large amount of bacterial protein was produced after AM fermentation, and its nutritional value was improved. FAM had significant effects on the growth performance of tiger grouper (p < 0.05). The high-density lipoprotein cholesterol (HDL-C) was highest in L4 group, being significantly different from L0 group. The area and diameter of hepatocytes were lowest in L3 and L4, and the density of hepatocyte was highest in L4 group and relatively decreased in L5 group. The mucosal height and muscular thickness were highest in L3 group. The intestinal microbiota structure of tiger grouper was changed under the intervention of FAM. The lower abundance of potential pathogenic bacteria and higher abundance of probiotics colonization in the L4 group showed that the dose of FAM had the best effect on improving the health of intestinal microbiota. This study indicates that the addition of FAM in the feed contributes to liver health, improves intestinal morphology, and regulates the intestinal microbiota of tiger grouper. The addition ratio of 1%–2% is better for intestinal and liver health, and a high addition ratio will cause liver damage. Our work will provide a reference for the addition and management of FAM in the aquaculture industry.

In order to reveal the acute toxicity and physiological changes of the spotted babylon (Babylonia areolata) in response to environmental manipulation, the spotted babylon was exposed to three pH levels (7.0, 8.0 and 9.0) of seawater and four concentrations of nitrite nitrogen (0.02, 2.7, 13.5 and 27 mg/L). The activities of six immunoenzymes, superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT), acid phosphatase (ACP), alkaline phosphatase (AKP) and peroxidase (POD), were measured. The levels of pH and nitrite nitrogen concentrations significantly impacted immunoenzyme activity over time. After the acute stress of pH and nitrite nitrogen, the spotted babylon appeared to be unresponsive to external stimuli, exhibited decreased vigor, slowly climbed the wall, sank to the tank and could not stand upright. As time elapsed, with the extension of time, the spotted babylon showed a trend of increasing and then decreasing ACP, AKP, CAT and SOD activities in order to adapt to the mutated environment and improve its immunity. In contrast, POD and GSH-PX activities showed a decrease followed by an increase with time. This study explored the tolerance range of the spotted babylon to pH, nitrite nitrogen, and time, proving that external stimuli activate the body’s immune response. The body’s immune function has a specific range of adaptation to the environment over time. Once the body’s immune system was insufficient to adapt to this range, the immune system collapsed and the snail gradually died off. This study has discovered the suitable pH and nitrite nitrogen ranges for the culture of the spotted babylon, and provides useful information on the response of the snail’s immune system.

Spotted babylon were exposed to three different pH levels (7.0, 8.0 and 9.0) and four different concentrations of ammonia nitrogen (0.02, 1.02, 5.10 and 10.20 mg/L) in seawater to determine their acute toxicity and physiological responses to environmental fluctuation. The study evaluated four antioxidant enzymes: catalase (CAT), alkaline, superoxide dismutase (SOD), peroxidase (POD) and glutathione peroxidase (GSH-PX), and two immunoenzymes: acid phosphatase (ACP) and phosphatase (AKP). Over time, the immunoenzyme activity was significantly affected by pH and ammonia nitrogen concentration. After being exposed to pH and ammonia nitrogen, the spotted babylon showed signs of unresponsiveness to external stimuli, reduced vitality, slow movement, and an inability to maintain an upright position. Over time, the spotted babylon exhibited a trend of increasing and then decreasing GSH-PX, CAT, and SOD activities to adapt to the changing environment and enhance its immunity. On the contrary, the POD and ACP activities exhibited a decreasing trend initially, followed by an increasing trend over time and the AKP activity showed a gradual increase with time. The combined effect of pH and ammonia was found to be stronger than the effect of either factor alone. The interaction between pH and ammonia increased the activity of the spotted babylon antioxidant enzymes, induced oxidative stress, and reduced the ability of the spotted babylon's non-specific immune system to reverse it. Thus, the reverse-back of the spotted babylon was higher when pH and ammonia stress were dual than when pH or ammonia were single-factor stresses. The study results will establish a theoretical basis for analyzing the risk of multiple factors to the spotted babylon, and also enrich the basic information about the shellfish immune system.

In cryopreservation technology, the choice of cryoprotectant plays a crucial role in cell survival and function. Different types of cryoprotectants, each with unique protective mechanisms, mitigate cellular damage from ice crystal formation during freezing. This study investigated the effects of different types and concentrations of cryoprotectants on the cryopreservation efficacy of noble scallop Mimachlamys nobilis sperm. Six cryoprotectants were tested, including four permeable cryoprotectants (dimethyl sulfoxide (DMSO), ethylene glycerol (EG), propylene glycerol (PG), methanol (MET)) and two non-permeable cryoprotectants (trehalose (TRE), fetal bovine serum (FBS)). The results showed that permeable cryoprotectants, which penetrate the cell membrane, regulate the osmotic pressure inside and outside cells to reduce dehydration damage. Among them, 10% DMSO provided the best protection, significantly preserving sperm motility, velocity, and morphology. Non-permeable cryoprotectants, although unable to penetrate cells, stabilized the extracellular environment at higher concentrations (such as FBS). Additionally, MET and FBS exhibited enhanced protective effects with increasing concentration, indicating their potential in reducing sperm structural damage at higher concentrations. Morphological observations indicated that freezing caused varying degrees of structural damage to sperm, with flagellar integrity being crucial for motility. Overall, selecting an appropriate cryoprotectant and concentration is essential for the efficient cryopreservation of M. nobilis sperm, providing a valuable reference for conserving germplasm resources of marine species.

The Asian seabass (Lates calcarifer) has high economic value and is the primary aquaculture species in China. Licorice (Glycyrrhiza uralensis) as a feed additive has demonstrated significant immunological benefits in aquaculture. However, its effects on the growth of aquatic animals are largely unexplored. This study explored the influence of licorice on the level of growth-related genes in Asian seabass by conducting an experiment using artificial feed with 0%, 1%, 3%, and 5% licorice. The impact on growth performance and the expression of several genes, including growth hormone–releasing hormone (GHRH), growth hormone (GH), growth hormone receptor (GHR), insulin-like growth factor 1 (IGF1), IGF2, IGF2 receptor (IGF2R), myostatin 1 (MSTN1), and myostatin 2 (MSTN2), were studied over 56 d. According to the results, the 3% and 5% licorice-supplemented diets significantly improved survival rates and weight gain compared to the control group. Licorice affected the level of growth-associated genes in Asian seabass and significantly increased the levels of GHR and IGF1 in the liver. However, a 5% licorice diet downregulated the expression of IGF2. As the licorice content in the diet increased, the levels of IGF2R and MSTN1 in the muscle tissue first decreased and then increased, and licorice addition inhibited the MSTN2 expression. The inclusion of licorice in the feed led to a significant downregulation of the GH and GHRH expression (p < 0.05). In summary, adding a certain proportion of licorice to the diet can improve the survival rate of the Asian seabass. Moreover, a proper proportion of licorice can increase the expression of related growth genes of fish, effectively increasing their weight gain rate and specific growth rate.