| 摘要: |
| 本研究利用MRS培养基，从山东东营刺参(Apostichopus japonicus)养殖池塘环境中分离到56株乳酸菌。以刺参腐皮综合征2种重要致病菌灿烂弧菌(Vibrio splendidus)和假交替单胞菌(Pseudoalteromonas nigrifaciens)为指示菌，采用牛津杯法测定所分离的乳酸菌对2株致病菌的抑制效果，筛选出1株具有较强抑菌活性的乳酸菌CSND-6，并对该细菌的形态、胞内产物和胞外产物的抑菌活性、生理生化实验、16S rDNA序列分析、菌株生长特性及对刺参安全性进行了研究。结果显示，菌株CSND-6对灿烂弧菌和假交替单胞菌的生长具有较好的抑制作用。其中，胞内产物的抑菌圈分别为17、22 mm，胞外产物的抑菌圈分别为25、31 mm。该菌株对刺参的高浓度浸浴胁迫实验结果表明，所分离的菌株对刺参是安全的。通过生理生化和16S rDNA序列分析表明，CSND-6属于乳杆菌科(Lactobacillaceae)，与副干酪乳杆菌(Lactobacillus paracasei JCM1171)的相似性为99.83%。菌株CSND-6在30℃~44℃、pH 为6~8范围内生长较快，24 h后进入对数生长期，28~32 h时达到生长高峰，最高值达2.10×109 CFU/ml。本研究筛选的副干酪乳杆菌属于刺参池塘养殖环境中土著益生菌，对刺参的主要病原菌抑制效果较好，并表现出了较好的生长特性，可为养殖池塘刺参疾病的生态防控和乳酸菌开发提供参考。 |
| 关键词: 刺参 腐皮综合征 副干酪乳杆菌 池塘养殖 |
| DOI：10.19663/j.issn2095-9869.20201216003 |
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| 基金项目: |
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| Isolation and biological characterization of Lactobacillus paracasei isolated from a sea cucumber pond culture system |
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LI Bin1, WANG Yingeng1, LIAO Meijie1, ZHANG Yonggang1, RONG Xiaojun1, WANG Jinjin1, YU Yongxiang1, ZHANG Zheng1, NING Luguang2
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1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of Sustainable and Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071;2.Shandong Haiyue Aquatic Science and Technology Co. Ltd, Dongying 257500
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| Abstract: |
| A total of 56 strains of Lactobacillus were isolated using MRS medium from a sea cucumber (Apostichopus japonicus) culture pond in Dongying, Shandong Province. Their antibacterial activities were analyzed using the Oxford cup method, and Vibrio splendidus and Pseudoalteromonas nigrifaciens were used as indicator bacteria. A Lactobacillus strain, CSND-6, showed strong antibacterial activity. The morphology, antibacterial activities of intracellular and extracellular components, physiological and biochemical tests, 16S rDNA sequence analysis, growth characteristics, and effects of strain CSND-6 on sea cucumbers were studied. The results showed that strain CSND-6 had a significant inhibitory effect on the growth of V. splendens and P. nigrifaciens. The inhibition zones of the intracellular components were 17 mm and 21 mm, and that of extracellular components were 25 mm and 33 mm for V. splendens and P. nigrifaciens, respectively. The safety analysis showed that the strain was safe for sea cucumbers. The results of the physiological and biochemical tests and 16S rDNA sequence analysis showed that the strain CSND-6 belongs to family Lactobacillaceae; the similarity between CSND-6 and Lactobacillus paracasei JCM1171 was 99.83%. The strain CSND-6 grew rapidly between 30℃ to 44℃ and in a pH between 6 to 8. It entered the logarithmic growth phase after 24 h and reached the highest concentration (2.10×109 CFU/ml) after 28~32 h. Since strain CSND-6 was isolated from the environment of the sea cucumber culture ponds and exerted strong inhibitory effects on the main pathogenic bacteria of sea cucumbers, it holds potential as an agent to prevent and control infections and diseases in sea cucumber. This study provides the foundation for the development and utilization of probiotics in sea cucumber cultures. |
| Key words: Apostichopus japonicus Skin ulcer syndrome Lactobacillus paracasei Pond culture |
