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为研究放线菌对海水养殖病原弧菌(哈维弧菌Vibrio harveyi和灿烂弧菌Vibrio splendidus)的抑制效果,从繁茂膜海绵Hymeniacidon perlevis中分离筛选出了一株具有抑菌活性的放线菌(FH),并采用16S r DNA基因序列进行分析,初步鉴定为球孢链霉菌Streptomyces globisporus;以抑菌活性为导向对放线菌FH菌株的发酵条件进行优化,并通过测量发酵沉淀菌丝体干质量,获得了FH菌株生长曲线,同时研究了发酵产物保存温度及时间对抑菌效果的影响,以及FH发酵产物在灭菌海水中对弧菌的抑制作用。结果表明:对FH菌株生长曲线测定结果显示,0~15 h为适应期,15~72 h为指数生长期,72~120 h为稳定期,120 h后为衰亡期;抑制弧菌效果试验显示,FH菌株发酵正交试验的最优条件为温度25℃、p H 7.5、装液量30m L(250 m L三角瓶);在正交试验最优条件下,FH菌株发酵培养72 h获得的粗提物抑制哈维弧菌和灿烂弧菌的效果最好;FH菌株发酵粗提物在4℃和-20℃下保存60 d和在室温保存7 d,其抑菌效果无显著性差异(P>0.05);在100 m L灭菌海水中,FH菌株发酵产物2 m L可有效抑制哈维弧菌和灿烂弧菌的繁殖。研究表明,海绵放线菌及其发酵产物在海水养殖领域具有潜在应用价值。
In order to study the inhibitory effect of Actinomyces on Vibrio harveyi and Vibrio splendidus, a strain of actinomycetes with antibacterial activity was isolated from hymeniacidon perlevis (FH). The 16S rDNA sequence was used for the analysis. The strain was initially identified as Streptomyces globisporus. The fermentation conditions of actinomycete strain FH were optimized by antibacterial activity. The growth curve of FH strain was obtained. The effect of fermentation temperature and time on the bacteriostasis and the inhibitory effect of FH fermentation on Vibrio parahaemolyticus in sterilized seawater were also studied. The results showed that the growth curve of FH strain showed that the adaptable period was 0-15 h, the exponential phase was 15-72 h, the stationary phase was 72-120 h and the decay stage was 120 h later. , The optimum conditions for FH strain fermentation orthogonal test temperature 25 ℃, p H 7.5, liquid volume 30m L (250 m L flask); orthogonal test under the optimal conditions, fermentation FH strains obtained 72 h The crude extract inhibited Vibrio harveyi and Vibrio sp. With the best results. The crude extracts of FH strain had no significant difference in antibacterial activity when stored at 4 ℃ and -20 ℃ for 60 days and at room temperature for 7 days P> 0.05). In the 100 m L sterilized seawater, the fermentation product of FH strain 2 m L could effectively inhibit the growth of V. harveyi and V. splendidum. Studies have shown that sponge actinomycetes and their fermentation products in the field of mariculture has potential applications.