两株潜在益生菌安全性评价及其对凡纳滨对虾免疫力和抗WSSV能力影响  

Safety Evaluation of two Probiotics Strains and Their Effects on the Immunity and the Ability of Anti-WSSV of Litopenaeus Vannamei

作　　者：王晓琳[1] 马甡[1] 单洪伟[1] 

机构地区：[1]中国海洋大学海水养殖教育部重点实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2016年第10期39-47,共9页Periodical of Ocean University of China

基　　金：山东省自然科学基金项目(ZR2014CP005);公益性行业(农业)科研专项项目(201103034)资助~~

摘　　要：本研究以2株潜在益生菌(芽孢杆菌(Bacillus.sp)BZ5株和溶藻弧菌(Vibrio alginolyticus)VZ5株)为研究对象。通过研究其溶血特性、常见抗生素的敏感性及感染菌体后凡纳滨对虾(Litopenaeus vannamei)的存活率,探讨了两菌株对凡纳滨对虾的安全性。注射和浸泡方式下,研究两菌株对凡纳滨虾的超氧化物歧化酶(SOD)、过氧化酶(POD)、抗菌酶活力变化的影响以及感染WSSV的凡纳滨对虾的存活率,探讨了两菌株对凡纳滨对虾免疫力和抗WSSV能力的影响。研究显示:芽孢杆菌BZ5株和溶藻弧菌VZ5株均未表现出溶血特征,且对多数常见抗生素敏感或中度敏感。在106cfu/mL浓度下,芽孢杆菌BZ5株和溶藻弧菌VZ5株未对凡纳滨对虾表现出明显的毒害作用。注射和浸泡方式下,芽孢杆菌组和溶藻弧菌组对虾的SOD、POD活力显著高于对照组(P<0.05),其中芽孢杆菌组对虾的SOD、POD活力显著高于溶藻弧菌组(P<0.05)。感染WSSV后,芽孢杆菌组和溶藻弧菌组对虾的半数致死时间比对照组分别延长了27.38%和21.43%(注射菌体)、24.73%和8.60%(浸泡菌体)。研究结果表明,芽孢杆菌BZ5株和溶藻弧菌VZ5株在提高对虾免疫力和抗WSSV能力方面具有较好的效果,在对虾养殖生产中有潜在的应用前景。Probiotics are widely used in aquaculture due to their excellent performance in promoting the growth and immunity of cultured aquatic animals,decreasing epidemic disease infection,and improving the culture environment.With the extension of their applications,safety problems aroused wide attentions.The purpose of this study was to evaluate the safety of Bacillus sp.BZ5 and V.alginolyticus VZ5 and reveale their effects on the immune-related enzyme activity and the ability of anti-WSSV of L.vannamei.This study included hemolysis,drug sensitivity,injection and immersion tests.In hemolysis test,strain BZ5 and VZ5were cultured in Luria-Bertan medium with a salinity of 30 for 24h,and then inoculated in blond agar for observation of hemolysis.In drug sensitivity test,a sensitive degree of strain BZ5 and VZ5to 20 kinds of common antibiotics were recorded.In injection test,strain BZ5 and VZ5（50μL,10^6 cfu/mL）were injected to shrimp muscle.In immersion test,shrimp were immersed in culture environment containing 10^6 cfu/mL of BZ5 and VZ5.The WSSV was injected to the shrimp muscle at the 4th of injection test and the 16 ^th of immersion test.Cumulative mortality and immune-related enzyme activity of shrimp were recorded during the injection test and immersion test.Strain BZ5 and VZ5did not cause a hemolysis phenomenon,and were sensitive or moderately sensitive to most of the common antibiotics.BZ5 and VZ5did not show toxic effect on L.vannamei at 10^6cfu/mL（in vivoinjection and immersion in bacterium solution）.In some studies,Vibrio alginolyticus was reported as a pathogen of shrimp.This may be related to the genetic diversity of V.alginolyticus strains resulting in greatly different virulence.The activity of SOD,POD of shrimp in Bacillus Sp.group and V.alginolyticus group were significantly higher than those of the control（P〈0.05）,and those in the Bacillus Sp.group were significantly higher than those in V.alginolyticus group（P〈0.05）.The strain BZ5 showed better performance because its metabolites could

关 键 词：芽孢杆菌 溶藻弧菌 凡纳滨对虾 安全性评价 免疫力 抗WSSV能力 

分 类 号：S968.22[农业科学—水产养殖]