本文選題：海綿共附生放線菌 + 抑菌活性�。� 參考：《大連海洋大學(xué)》2017年碩士論文

【摘要】：本文以研究海綿共附生放線菌次級代謝產(chǎn)物對海水養(yǎng)殖病原弧菌的抑菌活性為主要目的,從腎指海綿(Reniochalina sp.),山海綿(Mycale sp.)和居蟹皮海綿(Suberites domuncula)中分離培養(yǎng)出27株共附生放線菌,與實(shí)驗(yàn)室前期保存的海綿放線菌去重后共59株,16S r DNA序列比對初步鑒定均為鏈霉菌屬(Streptomyces)放線菌。以副溶血弧菌(Vibrio parahaemolyticus)、哈維弧菌(Vibrio harveyi)、鰻弧菌(Vibrio anguillarum)、燦爛弧菌(Vibro Splendidus)4株革蘭氏陰性菌和海豚鏈球菌(Streptococcus iniae)、馬胃葡萄球菌(Staphylococcus equorum)2株革蘭氏陽性菌為指示菌,對59株放線菌的抑菌活性進(jìn)行反復(fù)篩選,篩選出11株抑菌活性較強(qiáng)且具有廣譜性的放線菌,并確定海綿放線菌FH作為研究的目標(biāo)菌株。以副溶血弧菌、哈維弧菌為指示菌,對FH菌株的抑菌活性研究如下:對海綿放線菌FH的16S r DNA序列進(jìn)行測序分析,初步鑒定為球孢鏈霉菌(Streptomyces globisporus)。以抑菌活性為導(dǎo)向?qū)H菌株的發(fā)酵條件進(jìn)行了優(yōu)化,優(yōu)化結(jié)果為溫度25℃,p H值7.5,裝液量30 m L/250 m L,發(fā)酵時(shí)間120 h。FH菌株特征研究顯示,FH發(fā)酵液粗提物相當(dāng)于慶大霉素標(biāo)準(zhǔn)液的效價(jià)為387.27 mg/L,在4℃和-20℃低溫條件下保存2個(gè)月對各指示菌的抑菌活性穩(wěn)定。在滅菌海水中,添加FH菌株發(fā)酵產(chǎn)物的實(shí)驗(yàn)組,初始密度103個(gè)/m L的副溶血弧菌和哈維弧菌均得到了有效抑制甚至殺死;添加FH菌絲體的實(shí)驗(yàn)組中,初始密度103個(gè)/m L的副溶血弧菌和初始密度102個(gè)/m L的哈維弧菌得到了有效抑制。在海參育苗中,FH菌絲體在弧菌病害預(yù)防中則表現(xiàn)出較好的結(jié)果,作為生物防治具有可行性。而FH發(fā)酵產(chǎn)物和發(fā)酵液粗提物不適合直接添加到養(yǎng)殖海水中作為預(yù)防或治療的手段。上述海綿放線菌FH及其發(fā)酵產(chǎn)物的抑菌活性研究,說明了海綿放線菌在天然活性產(chǎn)物開發(fā)上具有重要潛力,在海水養(yǎng)殖病害防治領(lǐng)域具有廣闊的應(yīng)用前景。
[Abstract]:The aim of this paper is to study the antibacterial activity of secondary metabolites of actinomycetes sp.against pathogenic Vibrio vibrio in mariculture, from Reniochalina sp. of Reniochalina sp., Mycale sp. A total of 27 coepiphytic actinomycetes were isolated from Suberites domuncula and compared with 59 strains of actinomyces sponges stored in laboratory after being removed and identified as Streptomyces actinomycetes. Vibrio parahaemolyticus, Vibrio harveyii, Vibrio anguillarum, Vibrio Splendidus)4, Streptococcus iniaeae, Staphylococcus equorum)2, Gram-positive bacteria, The antimicrobial activity of 59 actinomycetes was screened repeatedly, 11 actinomycetes with strong antibacterial activity and broad spectrum were screened out, and the spongy actinomycetes FH was selected as the target strain. The antibacterial activity of Vibrio parahaemolyticus and Vibrio Harvey against FH strain was studied as follows: the 16s r DNA sequence of actinomyces sponges FH was sequenced and identified as Streptomyces globisporusus. The fermentation conditions of FH strain were optimized based on bacteriostatic activity. The optimized results were as follows: temperature 25 鈩，

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