自制环保酵素对水产病原菌的抑制效果及其发酵菌株的分离鉴定

Inhibitory activity of home-made garbage enzyme against aquatic pathogens and isolation and identification of its fermentation strains

为探讨自制环保酵素对水产病原微生物的抑制作用,该研究对其发酵菌株进行了分离。通过比浊法和平板划线法,确定自制环保酵素对水产病原微生物坎氏弧菌SP-1(Vibrio campbellii SP-1)、哈维氏弧菌SP-1(V.harveyi SP-1)、轮虫弧菌SP-1(V.rotiferianus SP-1)和嗜水气单胞菌245(Aeromonas hydrophila)的最低抑制浓度分别是体积分数是体积分数为11.5%、11%、6.2%和11%,最小杀菌浓度分别是体积分数为16%、16%、16%和12%。攻毒保护试验表明,自制环保酵素无菌上清液可使嗜水气单胞菌对斑马鱼(Danio rerio)的致死率降低(90.5±6.49)%。pH、温度、蛋白酶敏感性试验显示自制环保酵素的主要抗菌活性物质为酸性物质,对加热和蛋白酶处理稳定。进一步从环保酵素中分离主要发酵菌株,通过ITS序列分析鉴定为库德毕赤酵母(Pichia kudriavzevii)。研究结果可为自制环保酵素在水产养殖中对抗细菌性疾病的应用提供理论基础。

In order to evaluate the inhibitory activity of home-made garbage enzyme against aquatic pathogens and to isolate their fermentation strains,we determined the minimum inhibitory concentrations of the home-made garbage enzyme on the aquatic pathogenic microorganisms Vibrio candida SP-1,V.harveyi SP-1,V.rotiferianus SP-1 and Aeromonas hydrophila 245 as 11.5%,11%,6.2%and 11%in volume fraction by turbidimetric method.The minimum bactericidal concentrations were determined as 16%,16%,16%and 12%in volume fraction by flat line scribing method.The challenge test proves that the supernatant of home-made garbage enzyme decreased(90.5±6.49)%of the mortality in zebrafish induced by A.hydrophila.The sensitivity tests of pH,temperature and protease prove that the main antibacterial active substances of home-made garbage enzyme were acidic substances that were stable to heating and protease treatment.Furthermore,the primary fermentation strain was separated from the garbage enzyme and identified as Pichia kudriavzevii by ITS sequence analysis.The results provide a theoretical basis for applying home-made garbage enzyme in combating bacterial diseases in aquaculture.