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| 饲料中添加丁酸梭菌对刺参生长、免疫和消化的影响* |
| Effects of dietary Clostridium butyricum on the growth performance, non-specific immunity, and digestive ability of juvenile sea cucumber, Apostichopus japonicus |
| 投稿时间：2025-03-09 修订日期：2025-05-14 |
| DOI： |
| 中文关键词: 刺参 丁酸梭菌 生长 免疫 消化 |
| 英文关键词: Apostichopus japonicus Clostridium butyricum growth immunity digestive ability |
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| 中文摘要: |
| 丁酸梭菌(Clostridium butyricum)已广泛作为益生菌添加到饲料中以改善水产动物生长性能、饲料效率和免疫能力,但在刺参中的应用还鲜有报道。本实验旨在研究基础饲料中添加丁酸梭菌对刺参(Apostichopus japonicus)生长、免疫和消化指标的影响。选取初始体重为(5.92±0.11)g的刺参750头,随机分成5组,每组3个重复,每个重复50头。对照组(G0组)刺参投喂基础饲料,实验组在基础饲料中分别添加1×106CFU.g ?1、1×107CFU.g ?1、1×108CFU.g?1和1×109CFU.g?1的丁酸梭菌,记作G1、G2、G3、G4组,饲养周期为90d。结果表明：1)G4组刺参特定生长率最高,为1.01%/d,G3组和G4组显著高于其他组(P＜0.05),各组之间存活率没有显著差异(P＞0.05)；2)各实验组和对照组之间,体腔液细胞总数没有显著差异(P＞0.05),G4组体腔液细胞吞噬活性显著高于其他组(P＜0.05),G3组体腔液细胞吞噬活性显著高于对照组、G1、G2组(P＜0.05),G4组体腔液细胞呼吸爆发活力显著高于对照组和其他实验组(P＜0.05)；3)G3和G4组酚氧化酶、酸性磷酸酶和碱性磷酸酶活性显著高于其他三组(P＜0.05),溶菌酶活性显著高于G1组和对照组(P＜0.05)；4)G3组和G4组肠道蛋白酶、淀粉酶和脂肪酶活性显著高于G0和G1组(P＜0.05)。综上所述,饲料中添加丁酸梭菌可促进刺参生长,提高刺参免疫和消化能力。以生长、免疫和消化指标为评价标准,同时考虑到生产成本,建议刺参饲料中丁酸梭菌的添加水平为1×108CFU.g ?1。 |
| 英文摘要: |
| The production of juvenile sea cucumber is a vigorous industry in China, with an annual output of 50~60 billion individuals. Antibiotics was used to control the diseases caused by high intensity of seedling in hatchery, which can result in antibiotic residues in adult sea cucumber. Prophylactic administration of probiotics has emerged as a promising alternative strategy to control sea cucumber diseases by strengthening their immunity. Clostridium butyricum, an obligate anaerobic bacterium, is widely used as a feed additive in aquaculture to promote growth, improve immunity, and enhance digestive capacity in aquatic animals. However, its use in sea cucumber has rarely been reported. This work was conducted to evaluate the potential of C. butyricum as a dietary probiotic to improve growth, immunity and digestive capacity in the sea cucumber Apostichopus japonicus. Healthy juvenile sea cucumbers with an average body weight of (5.92±0.11)g were randomly divided into five groups with three replicates in each group, and 50 sea cucumbers in each replicate. The animals were reared in aquaria and fed diets supplemented with C. butyricum at concentrations of 0 CFU/g (control group)(G0), 106 CFU/g (G1), 107 CFU/g (G2), 108 CFU/g (G3), and 109 CFU/g(G4) for 90 days, respectively. During the experiment, water temperature was maintained at 11~23 ℃, salinity at 30~32, dissolved oxygen at a minimum of 6 mg/L, and pH at 7.8~8.2. At the end of the feeding trial, all sea cucumbers in each aquarium were weighed and counted. Five sea cucumbers from each aquarium were sampled, starved for 24h, and their coelomic fluid and intestine were collected for immune and digestive indices analysis, respectively. Coelomocyte counts were determined using a hemocytometer and expressed as cells per ml. Phagocytosis activity was assessed using a modified neutral red method, while intracellular reactive oxygen species (ROS) production was measured using the nitroblue tetrazolium (NBT) method. Phenoloxidase (PO) activity was determined spectrophotometrically using L-3, 4-dihydroxyphenylalanine (L-DOPA) as substrate and trypsin as elicitor. Acid phosphatase (ACP) and alkaline phosphatase (AKP) activity were measured using commercial kits (Nanjing Jiancheng, China). Lysozyme activity was measured by a spectrophotometric method based on the lysis of Micrococcus lysodeikticus. Digestive enzyme (amylase, trypsin, and lipase) activities were determined using enzyme activity assay kits (Nanjing Jiancheng, China) following the manufacturer’s instructions. Results showed that dietary supplementation with C. butyricum at concentrations of 108 CFU/g or higher significantly improved the specific growth rate and weight gain rate of sea cucumbers (P < 0.05). However, C. butyricum had no significant effect on survival rate of the sea cucumber (P＞0.05). Phagocytosis and respiratory burst activity were significantly improved in coelomocytes of sea cucumbers fed with C. butyricum at 109 CFU/g diet (P < 0.05). Additionally, phenoloxidase activity, acid phosphatase activity, and alkaline phosphatase activity in coelomocytes were significantly higher in G3 and G4 groups compared to the other three groups (P < 0.05). Lysozyme activity in coelomocytes was also significantly higher in G3 and G4 groups compared to the G1 and control groups (P < 0.05). Furthermore, C. butyricum also significantly improved the digestive enzyme activity of A. japonicus. Sea cucumbers in the G3 and G4 groups showed significantly higher amylase, trypsin and lipase activities compared to those in the G0 and G1 groups. The above results showed that C. butyricum as a feed additive can promote growth, activate the immune system, and enhance digestive ability in sea cucumber. The present study confirmed the potential beneficial effects of C. butyricum as dietary probiotic in juvenile A. japonicus, and the optimal supplemental level of C. butyricum is 108 CFU/g. These findings provide valuable insights for developing sustainable and antibiotic-free strategies in the sea cucumber industry. |
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