溶壁酶体外抗同丝水霉活性研究
In-Vitro Efficacy of Lywallzyme for Treating Saprolegnia monoica Pringsh in Fish
投稿时间:2016-09-28  修订日期:2018-01-22
中文关键词:溶壁酶  同丝水霉  最低抑菌浓度  抑菌效果
英文关键词:lywallzyme  Saprolegnia monoica  the minimum inhibitory concentration (MIC)  antifungal efficacy
基金项目:广东省科技计划项目(2013B020307003)
作者单位E-mail
华亚南 广东省微生物研究所广州 510070 广东碧德生物科技有限公司广州 510070 458782260@qq.com 
吴清洋 广东省微生物研究所广州 510070广东碧德生物科技有限公司广州 510070省部共建华南应用微生物国家重点实验室广州 510070广东省微生物应用新技术公共实验室广州 510070广东省菌种保藏与应用重点实验室广州 510070  
张玲敏 广东碧德生物科技有限公司广州 510070  
谭文俊 广东碧德生物科技有限公司广州 510070  
张丽 2. 广东省微生物研究所广州 510070广东碧德生物科技有限公司广州 510070 gdbide@163.com 
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中文摘要:
      为探究溶壁酶对水产动物水霉病的预防和治疗效果,试验以同丝水霉(Saprolegnia monoica )为研究对象,溶壁酶设0、5、10、20、40、80 mg/mL 共计6个浓度的处理组,取浓度为103个/mL的水霉孢子菌悬液0.1 mL,涂布于不同溶壁酶浓度的马铃薯葡萄糖琼脂培养基(PDA)上,25 °C培养5 d,以最低抑菌浓度为判定指标,研究溶壁酶溶解同丝水霉孢子的抑菌活性;在此基础上,以溶壁酶最低抑菌浓度30℃分别水浴2 h和4 h处理同丝水霉菌丝块后于PDA培养基上,25°C培养5 d,通过用体外抑菌试验、菌丝显微结构观察等方法,研究溶壁酶最低抑菌浓度对同丝水霉菌丝的抑菌活性,为进一步开发水产动物抗真菌(替代孔雀石绿)药物提供科学依据。结果显示,溶壁酶对同丝水霉孢子的最低抑菌浓度为20 mg/mL,通过间隔添加溶壁酶该浓度,72 h能够有效抑制同丝水霉菌丝的生长;溶壁酶30℃水浴处理同丝水霉菌丝块的时间(2 h和4h)与溶壁酶溶解菌丝效果成正相关
英文摘要:
      Saprolegnia monoica Pringsh is an oomycete, often referred to as a water mold, which causes saprolegniasis in freshwater fish. S. monoica is a eukaryotic microorganism that tolerates a broad temperature range and it is widely distributed in China. Lywallzyme is a lytic enzyme that can dissolve the cell walls of oomycetes, inhibiting growth and causing dieback. In this study, we investigated the activity of lywallzyme toward S. monoica by conducting in vitro stasis experiments and observing the mycelia under a microscope. The objectives were to determine the efficacy of lywallzyme for preventing and treating saprolegniasis in aquatic animals and to explore the potential of replacing malachite green with a safe, efficient and environmentally friendly treatment. Potato dextrose agar (PDA) was prepared at each of six lywallzyme concentrations (0, 5, 10, 20, 40, 80 mg/mL) and PDA plates were prepared in triplicate for each treatment. A suspension of S. monoica (0.1mL, 103 CFU/mL) was coated on the PDA plates and the plates were cultured for 5 days at 25 oC. The activity of lywallzyme in reducing S. monoica spores was observed and the minimum inhibitory concentration (MIC) of lywallzyme was obtained. Samples of S. monoica mycelia were then placed in a water bath (30 oC) for 2 or 4 hours at the MIC of lywallzyme. The treated mycelium were then transferred to PDA plates, cultured for 5 days at 25 oC, and the structure of S. monoica mycelia were observed under a microscope. Results show that the minimum effective concentration of lywallzyme for treatment of Saprolegnia is 20 mg/mL. Growth of S. monoica is effectively inhibited for 72 h by intermittent addition of 20 mg/mL lywallzyme [intermittent? Need to list the time interval]. Observations showed that lywallzyme ruptures S. monoica mycelium and inhibits S. monoica growth. The water bath at 30 oC promoted the activity of lywallzyme and strengthened the activity of lywallzyme and the effectiveness of lywallzyme increased with the time of water bath treatment.
华亚南,吴清洋,张玲敏,谭文俊,张丽.2018.溶壁酶体外抗同丝水霉活性研究[J].水生态学杂志,39(2):88-94.
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