摘要
近年来,急性肝胰腺坏死病(Acute hepatopancreatic necrosis disease,AHPND)造成了凡纳滨对虾(Litopenaeus vannamei)养殖业巨大的经济损失,其病原为一类含有致病基因PirAB的弧菌(Vibrio),主要为副溶血弧菌(Vibrio parahaemolyticus,VP_(AHPND)).本研究以VP_(AHPND)为受试菌株,从江苏海域捕获的黄姑鱼(Nibea albiflora)体内分离筛选对VP_(AHPND)具有抑制作用的拮抗菌株.通过形态学、生理生化及分子生物学方法进行鉴定,研究了其拮抗效果、拮抗活性物质,并对生物安全性和被动保护效果进行评估.结果表明,分离筛选出1株具有良好拮抗效果的菌株JSHY97,经鉴定该菌株为铜绿假单胞菌(Pseudomonas aeruginosa).该菌株对3株具有不同分型的VP_(AHPND)均具有良好的拮抗效果,其抑菌圈直径达19.25 mm~21.80 mm,初步判断其活性物质来源于胞外产物.口服JSHY97的凡纳滨对虾的7 d保护率也达到80%,拮抗组的凡纳滨对虾体内的毒力基因PirA拷贝数显著低于对照组(P<0.05).安全性实验表明JSHY97对斑马鱼(Brachydanio reriovar)和凡纳滨对虾无毒力.由此表明,本研究得到的铜绿假单胞菌菌株JSHY97对VP_(AHPND)具有较好的抑制作用,为AHPND的防治提供了新的菌株资源.
Acute hepatopancreatic necrosis disease causes serious economic losses to the Litopenaeus Vannamei shrimp industry in recent years.Studies have shown that the pathogen responsible for this disease is a type of Vibrio containing PirAB virulence factors,specifically Vibrio parahaemolyticus(VP_(AHPND)).In this paper,the antagonistic strain of VP_(AHPND)was isolated from Nibea albiflora in the coastal waters of Jiangsu province,with identified by morphological characteristics,physiological and biochemical characteristics,and 16S rDNA sequence analysis.Its antagonistic effect and antagonistic active substances were preliminarily studied,and the assessment of bio-safety were preliminarily optimized.The results showed that the strain JSHY97 with powerful antagonistic effect was screened and identified as Pseudomonas aeruginosa.JSHY97 had an antagonistic effect against 3 strains of VP_(AHPND)with a zone of inhibition diameter of 19.25 mm-21.80 mm,and its active substances were preliminarily judged to be extracellular products.After 7 days of diet,protection rates of 80%were also achieved by feeding JSHY97 to L.vannamei.The copy number of the virulence gene PirA in the JSHY97-fed L.vannamei was significantly lower than that in the control group(P<0.05).Safety experiments showed that JSHY97 was non-toxic to Brachydanio reriovar and L.vannamei.Pseudomonas aeruginosa strain JSHY97 has a good inhibitory effect on VP_(AHPND),which provides a new strain resource for the prevention and treatment of AHPND.
作者
刘学实
蒋葛
沈辉
乔毅
成婕
曹晓慧
窦雅斌
温梦莹
万夕和
王李宝
黎慧
史文军
Liu Xueshi;Jiang Ge;Shen Hui;Qiao Yi;Chen Jie;Cao Xiaohui;Dou Yabin;Wen Mengying;Wan Xihe;Wang Libao;Li Hui;Shen Wenjun(College of Marine Science and Engineering,Nanjing Normal University,Nanjing 210023,China;Jiangsu Institute of Oceanology&Marine Fisheries,Nantong 226007,China;College of Fisheries and Life,Shanghai Ocean University,Shanghai 201306,China)
出处
《南京师大学报(自然科学版)》
CAS
北大核心
2023年第1期73-79,87,共8页
Journal of Nanjing Normal University(Natural Science Edition)
基金
国家农业长期性基础性重大疫病监测项目(NAES-AH-011)
江苏省农业专项(2022-SJ-037-02)
中国盐城渔业高质量发展研究课题(YCSCYJ2021005)
南通市科技计划项目(JC2021089、JCZ20045、JC2020119、JC2020116)。
关键词
VP_(AHPND)
凡纳滨对虾
拮抗菌
铜绿假单胞菌
Vibrio parahaemolyticus
Litopenaeus vannamei
antagonistic bacterium
Pseudomonas aeruginosa