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應用NTG-UV複合誘變法開發弧菌拮抗菌Bacillus pumilus D5之突變菌株

  • 出版日期:107-12-31
  • 標題title(英):
    Using UV/NTG Mutagenesis to Develop Bacillus pumilus D5 Mutants with High Anti-Vibrio Abilities
  • 作者:朱惠真‧黃美瑩‧許晉榮‧張錦宜‧吳豐成‧鄧晶瑩
  • 作者auther(英):Huei-Jen Ju, Mei-Ying Huang, Jinn-Rong Hseu, Chin-I Chang, Feng-Cheng Wu and Ching-Ying Deng
  • 卷別:26
  • 期別:2
  • 頁碼:21-31

不論是野生魚種或是養殖魚種,弧菌性疾病一直以來都是世界性的重大威脅,也是高密度養殖發展的重要限制因子。本研究之弧菌拮抗菌是分離自白蝦 (Litopenaeus vannamei) 的短小芽孢桿菌D5 (Bacillus pumilus D5) 菌株,具有抑制腸炎弧菌、溶藻弧菌及哈維氏弧菌的能力。為了提高B. pumilus D5菌株的抑菌效果,本試驗利用紫外線突變法、亞硝基弧 (Nitrosoguanidine,NTG) 突變法以及紫外線與亞硝基弧共同誘變法突變B. pumilus D5菌株,目標是得到抗弧菌能力優於B. pumilus D5之突變菌株。突變的結果顯示:在三次篩選過程中最後得到四種突變細菌B. pumilus NG24、B. pumilus NG25、B. pumilus UV35及B. pumilus UV37,其對腸炎弧菌及哈維氏弧菌的抑菌圈直徑均和B. pumilus D5有顯著性差異 (p < 0.05)。在共培試驗中,在腸炎弧菌和B. pumilus D5或其突變菌株濃度為102 CFU ml-1與107 CFU ml-1的比例培養48 hr後,發現各拮抗菌均有抑制弧菌生長的能力 (p < 0.05),其中又以NG25的效果最佳。總括來說,B. pumilus D5和其突變菌之接種濃度需高於弧菌105倍才可以抑制腸炎弧菌的生長,期許能應用本突變菌株控制養殖池水弧菌生長量進而提高本拮抗菌之發展潛力。

摘要abstract(英)


Vibriosis, a collective Vibrio infection, is a widespread epizootic disease that affects most wild and farmed fish species worldwide, and is currently the major limiting factor in the development of intensive mariculture industry. In our previous study, we found that a Bacillus pumilus strain, D5, isolated from white shrimp (Litopenaeus vannamei) exhibited antibacterial ability against Vibrio alginolyticus, V. harveyi and V. parahaemolyticus. In order to get mutants with higher antibacterial abilities, the B. pumilus D5 strain was irradiated with ultraviolet (uv) radiation and treated with nitrosoguanidine (NTG). After three screenings, four mutant strains named B. pumilus mutant NG24, NG25, UV35 and UV37 were selected. The experimental results showed that the average bacteria inhibition zones against V. harveyi and V. parahaemolyticus of these four mutant strains provide significantly higher inhibition than B. pumilus D5 (p<0.05). In a co-culture test, the B. pumilus D5 strain and its mutants suppressed the growth of V. parahaemolyticus in vitro only if the initial inoculum of the given strain was up to 100000 times higher than the inoculum of V. parahaemolyticus. Furthermore, B. pumilus NG25 showed higher antagonistic abilities against pathogenic V. harveyi and V. parahaemolyticus than B. pumilus D5 (p<0.05). These study results reveal that the B. pumilus D5 mutants have potential applications for controlling pathogenic Vibrio spp. in aquaculture practices.