摘要
针对引起凡纳滨对虾高致死率的致急性肝胰腺坏死病副溶血弧菌(VpAHPND),生物拮抗是一种环保、有效、可持续的防治方式。本研究从山东东营对虾工厂化养殖场的尾水中取样,采用平板对峙法和打孔法筛选出目标菌株后,通过16SrDNA、gyrB-ropD基因序列比对的方式进行菌种鉴定。使用打孔法评估目标菌株拮抗VpAHPND能力,通过抑菌物质定位和共培养实验初步探究其抗VpAHPND机制。使用浸泡法测定该菌株对PL12虾苗的生物安全性,通过在水体添加拮抗菌养殖实验探究其对凡纳滨对虾存活率和弧菌数量的影响。结果表明,一株假交替单胞菌Pseudoalteromonas sp.strain 1125(以下简称Ps1125)具有良好的抑制VpAHPND能力,抑菌作用需要菌体活细胞接触,与上清液和死亡菌体无关。Ps1125对PL12的虾苗具有安全性。Ps1125加入水体养殖前期,具有降低养殖水体和对虾肝胰腺内的弧菌数量的效果。攻毒前期显著降低对虾肝胰腺弧菌数量,攻毒后期观察到对虾存活率显著提高。本实验证明一株海洋来源的假交替单胞菌Pseudoalteromonas sp.strain 1125通过活细胞接触抑制抗菌,通过养殖水体添加的方式降低了对虾肝胰腺内弧菌数量、水体弧菌数量,在凡纳滨对虾副溶血弧菌生物防控中具有良好的前景。
Biological antagonism is an effective,environmentally friendly,and sustainable method for the prevention and treatment of Vibrio parahaemolyticus(VpAHPND)infection,which causes acute hepatopancreatic necrosis disease in Litopenaeus vannamei.This study collected samples from seawater used for the industrial aquaculture of shrimp in Dongying City,Shandong,China.The target strains were screened using plate confrontation and drilling methods and were identified using 16SrDNA and gyrB-ropD gene sequence alignment.The target strain’s ability to antagonize VpAHPND was evaluated using the drilling method,and the mechanism of its resistance to VpAHPND was preliminarily explored through the localization of antibacterial compounds and co-culture experiments.A soaking method was used to determine the biological safety of this strain in post-larva12 shrimp.The effect of adding this antagonistic bacterium to water on the survival rate of L.vannamei and the number of Vibrio spp.was explored through aquaculture experiments.The results showed that Pseudoalteromonas spp.strain 1125(hereafter referred to as Ps1125)had a good ability to inhibit VpAHPND,with the antibacterial effect requiring contact with live cells,independent of the supernatant and dead cells.Ps1125 is safe for shrimp post-larva12,and adding it to the water during the early stage of aquaculture reduced the number of Vibrio spp.in the aquaculture water and shrimp hepatopancreas.The number of Vibrio spp.in the shrimp hepatopancreas was markedly reduced in the early stage of the challenge,whereas a significant increase in the shrimp survival rate was observed in the later stage.Results show that Ps1125 from marine sources inhibits antibacterial activity through live cell contact.When added to aquaculture water,the number of Vibrio spp.in the shrimp hepatopancreas and Vibrio spp.in the water can be suppressed.Thus,Ps1125 has excellent application prospects for the biological control of V.parahaemolyticus in L.vannamei.
作者
李嘉鑫
王宝杰
刘梅
蒋克勇
王雷
LI Jiaxin;WANG Baojie;LIU Mei;JIANG Keyong;WANG Lei(CAS Key Laboratory of Experimental Marine Biology,Institute of Oceanology,Chinese Academy of Sciences,Qingdao 266071,China;China Center for Ocean Mega-Science,Chinese Academy of Sciences,Qingdao 266071,China;University of Chinese Academy of Sciences,Beijing 100049,China;Marine Science Research Institute of Shandong Province,Qingdao 266104,China)
出处
《海洋科学》
北大核心
2025年第3期93-106,共14页
Marine Sciences
基金
山东省重点研发项目(2023TZXD047)
山东省自然科学基金(ZR2023MC096)
福建省中国科学院STS计划配套项目(2024T3028)。
