Department of Microbiology, Faculty of Agriculture, Cairo University, Giza 12613, Egypt.
AMB Express. 2012 Jan 26;2:9. doi: 10.1186/2191-0855-2-9.
In the present study, two of the most toxic bacterial strains of Bacillus sphaericus against mosquito were identified with the most recent genetic techniques. The PCR product profiles indicated the presence of genes encoding Bin A, Bin B and Mtx1 in all analyzed strains; they are consistent with protein profiles. The preliminary bioinformatics analysis of the binary toxin genes sequence revealed that the open reading frames had high similarities when matched with nucleotides sequence in the database of other B. sphaericus strains. The biological activity of B. sphaericus strains varied according to growing medium, and cultivation time. The highest yield of viable counts, spores and larvicidal protein were attained after 5 days. Poly (P) medium achieved the highest yield of growth, sporulation, protein and larvicidal activity for all tested strains compared to the other tested media. The larvicidal protein produced by local strains (B. sphaericus EMCC 1931 and EMCC 1932) in P medium was more lethal against the 3rd instar larvae of Culex pipiens than that of reference strains (B. sphaericus 1593 and B. sphaericus 2297). The obtained results revealed that P medium was the most effective medium and will be used in future work in order to optimize large scale production of biocide by the locally isolated Bacillus sphaericus strains.
在本研究中,使用最新的遗传技术鉴定了两种对蚊子最具毒性的球形芽孢杆菌菌株。PCR 产物图谱表明,所有分析的菌株中都存在编码 Bin A、Bin B 和 Mtx1 的基因;它们与蛋白质图谱一致。二元毒素基因序列的初步生物信息学分析表明,开放阅读框与数据库中其他 B. sphaericus 菌株的核苷酸序列具有高度相似性。菌株的生物活性根据生长培养基和培养时间而有所不同。在 5 天后,活菌数、孢子和杀幼虫蛋白的产量达到最高。与其他测试的培养基相比,聚(P)培养基对所有测试的菌株都实现了最高的生长、产孢、蛋白和杀幼虫活性产量。与参考菌株(B. sphaericus 1593 和 B. sphaericus 2297)相比,在 P 培养基中由本地菌株(B. sphaericus EMCC 1931 和 EMCC 1932)产生的杀幼虫蛋白对库蚊的 3 龄幼虫更具致死性。研究结果表明,P 培养基是最有效的培养基,将在未来的工作中用于优化由本地分离的球形芽孢杆菌菌株大规模生产生物杀灭剂。