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红笛鲷免疫器官发育组织学观察

广东海洋大学学报 2024年第2期44卷 124-129页

作者：赖钰秋李珠蔡双虎简纪常广东海洋大学水产学院/广东省水产动物病害防控与健康养殖重点实验室广东湛江524088

【目的】研究红笛鲷(Lutjanus sanguineus)免疫器官发育,为其健康养殖和疾病预防提供理论基础。【方法】从红笛鲷卵出膜开始取样,至出膜62 d,通过石蜡连续切片,H-E染色,进行红笛鲷3个主要免疫器官胸腺、头肾、脾脏发育的组织学研究。【... 详细信息

【目的】研究红笛鲷(Lutjanus sanguineus)免疫器官发育,为其健康养殖和疾病预防提供理论基础。【方法】从红笛鲷卵出膜开始取样,至出膜62 d,通过石蜡连续切片,H-E染色,进行红笛鲷3个主要免疫器官胸腺、头肾、脾脏发育的组织学研究。【结果与结论】红笛鲷出膜1 d时出现前肾,3 d肾管之间可见造血干细胞;出膜18 d,胸腺和头肾间出现细胞桥,头肾淋巴化;出膜62 d,肾小球仍存在于头肾中,表明头肾既是淋巴器官也是泌尿器官。红笛鲷出膜8 d时出现脾脏原基和红细胞发育;出膜22 d时观察到淋巴细胞;出膜24 d时淋巴细胞显著变多。胸腺是最后一个出现的器官,红笛鲷出膜12 d时出现胸腺原基;15 d时发现胸腺淋巴化。

关键词：红笛鲷胸腺头肾脾脏组织学

水生动物病原菌Ⅵ型分泌系统(T6SS)及其溶血素共调节蛋白研究进展

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大连海洋大学学报 2024年第1期39卷 162-171页

作者：伍水龙黄瑜王蓓汤菊芬蔡佳简纪常广东海洋大学水产学院广东湛江524088 湛江中心人民医院临床医学研究所广东湛江524045 广东省水产经济动物病原生物学及流行病学重点实验室广东湛江524088

Ⅵ型分泌系统(typeⅥsecretion system,T6SS)是多种革兰氏阴性菌编码的蛋白分泌装置,在毒力因子释放、生物被膜形成、铁离子摄取、囊泡运输、环境压力适应性及细菌胞内存活等方面发挥着重要功能,在副溶血弧菌(Vibrio parahaemolyticus)... 详细信息

Ⅵ型分泌系统(typeⅥsecretion system,T6SS)是多种革兰氏阴性菌编码的蛋白分泌装置,在毒力因子释放、生物被膜形成、铁离子摄取、囊泡运输、环境压力适应性及细菌胞内存活等方面发挥着重要功能,在副溶血弧菌(Vibrio parahaemolyticus)、溶藻弧菌(Vibrio alginolyticus)、哈维氏弧菌(Vibrio harveyi)、嗜水气单胞菌(Aeromonas hydrophila)和迟钝爱德华氏菌(Edwardsiella tarda)等水生动物病原菌的致病过程中发挥着关键作用。然而,有关水生动物病原菌T6SS组分及其功能的报道还比较匮乏。本文对几种水生动物病原菌T6SS的生物学功能与调控机理,以及T6SS关键调控因子溶血素共调节蛋白Hcp的系统进化关系与功能等最新研究情况进行了综述,并在水生动物病原菌T6SS的生物学功能、T6SS活性调控与环境因素的关联性、T6SS与致病菌代谢之间的调控关系等方面提出未来研究建议,以期为进一步开展水生动物致病机制研究及水产养殖细菌病的防控提供新思路。

关键词： Ⅵ型分泌系统水生动物病原菌溶血素共调节蛋白系统进化树功能位点

Construction of Case Database for Postgraduate Course Aquatic Animal Pathogen Biology

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Asian Agricultural Research 2022年第2期14卷 66-68页

作者： Huanying PANG Miao XIE Shuanghu CAI Ziling WANG Shiping YANG Yucong HUANG jichang jian College of Fisheries Guangdong Ocean UniversityZhanjiang 524088China National Demonstration Center for Experimental Fisheries Science Education Zhanjiang 524088China

As an application instructional course for Aquatic Animal Medicine(AAM),Aquatic Animal Pathogen Biology needs to be guided by a large number of examples and cases,but the current case database construction faces many urgent problems.In view of this,this paper analyzes the characteristics of professional education of AAM postgraduate students.With the goal of cultivating applied personnel that meet the requirements of the times,and the guiding ideology of strengthening the reform of the education model in colleges and universities,and improving the quantity and quality of personnel training,it builds a case database for the course Aquatic Animal Pathogen Biology in accordance with modern postgraduate teaching needs.

关键词： Case database Aquatic Animal Pathogen Biology Aquatic Animal Medicine(AAM)discipline

Study on Case Library Construction of Inspection and Quarantine of Aquatic Animal

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Asian Agricultural Research 2022年第9期14卷 64-66页

作者： Huanying PANG Miao XIE Shuanghu CAI Yucong HUANG Qianyi CHEN jianzhou DOU jichang jian College of Fisheries Guangdong Ocean UniversityZhanjiang 524088China National Experimental Teaching Demonstration Center of Fishery Science and Technology Zhanjiang 524088China Guangzhou Huibiao Testing Technology Center Guangzhou 510663China

By making use of the technical advantages of Guangzhou Huibiao Testing Technology Center in the field of testing, the project analyzes the characteristics of undergraduate professional education in aquatic animal medicine, and intends to build a case library of Inspection and Quarantine of Aquatic Animal suitable for modern undergraduate teaching needs, with training applied talents to meet the requirements of the times as the goal, and strengthening the reform of college education mode and improving the quantity and quality of talent training as the guiding ideology.

关键词： Case library Inspection and Quarantine of Aquatic Animal Aquatic animal medicine

Reform and Exploration of Integrating"Service Learning"into the Teaching of Pathogenic Biology of Aquatic Animals to Help Rural Revitalization

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Asian Agricultural Research 2023年第1期15卷 58-61页

作者： Huanying PANG Miao XIE Shuanghu CAI Ziling WANG Shiping YANG Yucong HUANG jichang jian College of Fisheries Guangdong Ocean UniversityZhanjiang 524088China National Experimental Teaching Demonstration Center of Fishery Science and Technology Zhanjiang 524088China

This paper intends to combine the development trend of the subject,restructure and optimize the course content,and construct the service learning model of Pathogenic Biology of Aquatic Animals for postgraduates.At the same time,through service practice,it is needed to further consolidate curriculum knowledge and skills,stimulate the learning initiative and enthusiasm of postgraduates,expand professional knowledge,improve professional quality,and lay a solid foundation for serving the national rural revitalization strategy in the future.

关键词： Pathogenic Biology of Aquatic Animals Service learning Rural revitalization

Construction of Bilingual Teaching Mode of Aquatic Animal Bacteriology in the Context of the Belt and Road Initiative

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Asian Agricultural Research 2022年第2期14卷 62-65页

作者： Huanying PANG Miao XIE Shuanghu CAI Ziling WANG Yucong HUANG Yu HUANG jichang jian College of Fisheries Guangdong Ocean UniversityZhanjiang 524088China National Demonstration Center for Experimental Fisheries Science Education Zhanjiang 524088China

Aquatic Animal Bacteriology is a basic course of Aquatic Animal Medicine discipline.Bacteriology is not only a frontier subject in the field of life sciences,but also one of the most rapidly developing scientific fields today.The Belt and Road Initiative has brought opportunities and challenges to the upgrading and development of this discipline,so this discipline faces many problems that need to be solved.In view of this,this study combined the development trend of the discipline,reorganized and optimized the course content,and prepared sophisticated multimedia courseware,to stimulate students'learning initiative and enthusiasm,expand the field of professional knowledge,improve their English translation and writing skills,so as to lay a solid foundation for their future service to the Belt and Road Initiative.

关键词： The Belt and Road Initiative Aquatic Animal Bacteriology Aquatic Animal Medicine discipline

GCRV 096 VP6 protein and its impacts on GCRV replication with different genotypes in CIK cells

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Aquaculture and Fisheries 2018年第5期3卷 184-190页

作者： Xiuying Yan Lingfang Xiong Jie Li Ya Wang Zaohe Wu jichang jian Yu Ding Guangdong Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals Guangdong Ocean UniversityZhanjiangGuangdong524088China

Grass carp reovirus(GCRV)is the causative agent of grass carp hemorrhagic disease,which has detrimental effects on the grass carp aquaculture industry.There are four known genotypes of GCRV and strains of different GCRV genotypes differ greatly.In this study,the diversity of the protein VP6 from different GCRV stains and the effect of genotype GCRV 096 on replication was investigated in CIK cells.Our results showed that the VP6 protein of GCRV 096(genotypeІ)exhibited limited homology to that of GCRV GD108(genotypeШ),with few residues conserved in predicted protein-protein interaction domains.GCRV 096 VP6 protein was expressed and purified and an antiserum against it was characterized.Addition of purified VP6 protein or antiserum to culture media of CIK cells inhibited the replication of GCRV 096 in these cells.In contrast,replication of GCRV GD108(genotypeШ)was not affected in CIK cells under the same condition.Overall,our results indicated that the protein VP6 and VP6 antiserum did not provide cross-protection against GCRV strains and this can be attributed to differences among GCRV genotypes.It will be important to consider multiple GCRV genotypes in the development of effective GCRV vaccines and other therapies against grass carp hemorrhagic disease.In addition,bioinformatics analysis also suggested that the protein VP6 may have a role in the process of GCRV infection.This study lays the foundation for the prevention of grass carp hemorrhagic disease and further detailed studies on the pathogenesis of GCRV.

关键词： Grass carp reovirus(GCRV) Protein VP6 Replication Functional analysis

草鱼miR-1260对鱼淋巴囊肿病毒中国株复制的调控作用

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广东海洋大学学报 2024年第3期44卷 25-33页

作者：马嘉霖战盈瑾周杰黄鉴涛闫秀英简纪常广东海洋大学水产学院/广东省水产动物病害防控与健康养殖重点实验室/水产经济动物病害控制广东普通高校重点实验室广东湛江524088

【目的】鉴定草鱼(Ctenopharyngodon idella)miR-1260(cid-miR-1260),分析cid-miR-1260在鱼淋巴囊肿病毒中国株(Lymphocystis disease virus China,LCDV-cn)感染草鱼卵巢细胞系(Grass carp ovary cells,GCO)过程中的时序表达特性,揭示cid-miR-1260对其靶基因znf574和LCDV-cn复制的调控作用。【方法】制备LCDV-cn感染的GCO细胞,通过茎环RT-PCR和测序鉴定cid-miR-1260;采用定量PCR检测cid-miR-1260在LCDV-cn感染GCO过程中的时序表达;应用生物信息学方法预测cid-miR-1260的靶基因,并用双荧光素酶报告基因系统验证靶基因;通过转染cid-miR-1260 mimic和cid-miR-1260 inhibitor使cid-miR-1260过表达和抑制表达,并用定量PCR检测cid-miR-1260、靶基因znf574、LCDV-cn mcp基因的表达;应用Reed-Muench法测定LCDV-cn在GCO细胞中的滴度。【结果】cid-miR-1260与其他已知miR-1260仅有一个碱基差异,且cid-miR-1260在LCDV-cn感染GCO过程中呈先上升后下降的表达变化,72 h时表达量最高(P<0.05);生物信息学分析表明,znf574为cid-miR-1260的靶基因;重组质粒pmirGLO-znf574-WT与cid-miR-1260 mimic共转染后,荧光素酶活性显著降低(P<0.05),证实znf574为cid-miR-1260的靶基因;cid-miR-1260过表达后,靶基因znf574的表达显著下降(P<0.05),LCDV-cn mcp的表达和LCDV-cn在GCO细胞中的滴度显著上升(P<0.05);抑制cid-miR-1260表达后,靶基因znf574的表达量显著上升(P<0.05),LCDV-cn mcp的表达量和LCDV-cn在GCO细胞中的滴度显著下降(P<0.05)。【结论】LCDV-cn感染GCO细胞后,cid-miR-1260的表达显著上调;在LCDV-cn感染中,cid-miR-1260对其靶基因znf574的表达起负调控作用,且cid-miR-1260表达与LCDV-cn的复制呈正相关,起促LCDV-cn感染作用,而锌指蛋白ZNF574具有抗LCDV-cn作用。本研究为淋巴囊肿病的防治提供新的依据和思路。

关键词：鱼淋巴囊肿病毒中国株草鱼卵巢细胞系 cid-miR-1260 锌指蛋白基因znf574 病毒复制

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多纹钱蝶鱼溃疡病病原的分离鉴定及药敏试验

大连海洋大学学报 2024年第3期39卷 392-401页

作者：阙莉文杨林狄涂祖豪蔡双虎黄郁葱简纪常广东海洋大学水产学院广东湛江524088 广东省水产动物病害防控与健康养殖重点实验室水产经济动物病害控制广东省普通高等学校重点实验室广东湛江524088

为研究2023年广西壮族自治区钦州湾网箱养殖多纹钱蝶鱼(Selenotoca multifasciata)发生溃疡病的原因,从患病多纹钱蝶鱼的肝、脾和肾组织中分离纯化出优势菌株,并对其进行形态学观察、生理生化特性鉴定、16S rRNA与dnaJ基因序列测序分析... 详细信息

为研究2023年广西壮族自治区钦州湾网箱养殖多纹钱蝶鱼(Selenotoca multifasciata)发生溃疡病的原因,从患病多纹钱蝶鱼的肝、脾和肾组织中分离纯化出优势菌株,并对其进行形态学观察、生理生化特性鉴定、16S rRNA与dnaJ基因序列测序分析、毒力基因检测、人工感染试验及药敏试验。结果表明:分离获得的优势菌在TCBS平板上形成黄色、圆形的光滑菌落,在TSA平板上形成乳白色微凸圆润菌落;生理生化特征分析显示,菌株为β溶血的革兰氏阴性菌,可发酵葡萄糖、蔗糖、甘露糖和阿拉伯糖,V-P试验呈阳性;基于16S rRNA和dnaJ基因序列构建的系统发育树显示,3株优势菌均与鳗弧菌(Vibrio anguillarum)聚为一支,综合鉴定表明菌株QZ991、QZ992和QZ993均为鳗弧菌;毒力基因检测显示,3株优势菌均携带virA、vah1、virC、empA、angM、flaA和ompU 7种毒力基因;人工感染试验显示,QZ991、QZ992和QZ993对多纹钱蝶鱼的半致死浓度(LD_(50))分别为5.10×10^(5)、4.33×10^(5)、2.45×10^(6)CFU/mL;药敏试验显示,3株优势菌均对多粘菌素B、庆大霉素、四环素类、喹诺酮类和多数头孢烯类等多种抗菌药物敏感;组织病理切片观察发现,患病多纹钱蝶鱼多脏器发生了不同程度的损伤,主要表现为充血、出血、坏死和炎性细胞浸润等。研究表明,鳗弧菌为广西钦州湾地区多纹钱蝶鱼溃疡病的病原,对多纹钱蝶鱼具有较强的致病性,并具有一定的耐药性,建议交替使用水产用多西环素和恩诺沙星进行治疗。

关键词：多纹钱蝶鱼鳗弧菌分离鉴定毒力基因药敏试验