中国鲎室内人工繁育和鲎卵、幼体常见病原鉴定及其防治  

Artificial breeding of Tachypleus tridentatus indoors and the identification and control of common pathogens for fertilized egg and larvae

作　　者：钟赟民 翁朝红[1] 王志婷 李佳怡 谢仰杰[1] ZHONG Yunmin;WENG Zhaohong;WANG Zhiting;LI Jiayi;XIE Yangjie(Ministry of Agriculture and Rural Affairs,Key Laboratory of Healthy Mariculture for the East China Sea,Fisheries College of Jimei University,Xiamen 361021,China)

机构地区：[1]集美大学水产学院,农业农村部东海海水健康养殖重点实验室,福建厦门361021

出　　处：《中国水产科学》2023年第6期799-811,共13页Journal of Fishery Sciences of China

基　　金：深圳质兰公益基金项目(2021040121B,2022070771B);厦门市海洋与渔业研究所合作项目(S23013)。

摘　　要：为了鉴定中国鲎(Tachypleus tridentatus)人工繁育过程中感染受精卵和低龄幼体的细菌或真菌等病原,同时探索防治措施,本研究开展了中国鲎在自然受精条件下的育苗实验。采用16S r DNA和ITS测序技术分别对感染病原的中国鲎受精卵表面细菌和真菌进行鉴定和微生物群落组成分析,同时选择溴氯海因溶液做为消毒剂开展不同浓度溴氯海因溶液对中国鲎1龄幼体的急性毒性研究,并根据安全浓度探索和确定适用于鲎卵和幼体的有效消毒浓度。结果表明,4对亲鲎共产卵16493粒,平均受精率66.25%,平均孵化率52.25%,病原感染率为47.75%,感染病菌的鲎卵死亡率100%,最终得到5704只1龄幼体。感染病原的中国鲎受精卵表面优势细菌为希瓦氏菌属(Shewanella)、海王星杆菌属(Neptuniibacter)、脱硫杆菌属(Desulfobacter)、寺崎菌属(Terasakiella)和哈拉科杆菌属(Halarcobacter),相对丰度分别为23.67%、7.21%、6.64%、5.41%和5.16%;优势真菌为肉座菌科(Hypocreaceae)的一未知种和曲霉菌属(Aspergillus),相对丰度为81.81%和10.48%。溴氯海因对1龄幼鲎的96 h半致死浓度为945.06 mg/L,安全浓度为94.51 mg/L,在安全浓度下溴氯海因对鲎卵孵化和1龄幼鲎培育有效消毒浓度为30~90 mg/L,此浓度范围内能够显著提高鲎卵孵化率(P<0.05),并降低鲎卵和1龄幼鲎的死亡率(P<0.05)。研究结果显示溴氯海因可有效降低受精卵和幼体被细菌、真菌等微生物感染的风险,可用于中国鲎人工繁育过程中的日常消毒。Chinese horseshoe crab(Tachypleus tridentatus)belongs to the Arthropoda,Chelicerata,and Merostomata taxonomic groups and is,specifically,a member of the horseshoe crab family(Limuroidea).Tachypleus amebocyte lysate(TAL)reagent,prepared from the hemolytic solution of T.tridentatus,can quickly coagulate into a colloid with bacteria;therefore,TAL is a conventional reagent for the detection of endotoxins.Due to habitat destruction and overfishing,T.tridentatus has become an endangered marine animal,with its resources on the verge of depletion.Artificial breeding and release are important ways to increase its population in the wild.Artificial breeding of T.tridentatus has been conducted as early as the 1980 s.However,artificial breeding strategies involve killing horseshoe crabs to obtain eggs,with few reports on larval cultivation;therefore,this breeding process still lacks detailed technical standards and specifications.In recent years,artificial breeding has been adopted to promote natural spawning of broodstock horseshoe crabs under simulated wild conditions to reduce the damage to these rare broodstock crabs;however,their corresponding hatchability and survival rates are relatively low.With limited artificial space indoors,high-density culture leads to an extremely small living space for fertilized eggs and larvae;therefore,these eggs and larvae become susceptible to infection by various bacteria,fungi,viruses,parasites,and other pathogens.Therefore,to improve the hatchability rate of fertilized eggs and the survival rate of larvae for large-scale artificial breeding,it is necessary to address these issues.In this study,nutrient fortification of the broodstock horseshoe crabs was initially conducted,followed by natural fertilization to obtain fertilized eggs for hatching.Then,the eggs infected with bacteria were collected to study the characteristics of different colored eggs.Next,16 s rDNA and ITS sequencing technology were used to identify bacteria and fungi on the surface of T.tridentatus fertilized eggs,and t

关 键 词：中国鲎 人工繁育 病原鉴定 溴氯海因 受精卵 幼体 

分 类 号：S941[农业科学—水产养殖]