虾肝肠胞虫感染病理特征、18S rRNA基因序列及系统进化树分析
PDF下载 (258)周 颖,祝 波,钱 冬,卢先东,李小冰,李 政,刘艳红,蔡惠凤,吴仲宁,袁思平,张崇虎,叶乐平,马荣荣*.虾肝肠胞虫感染病理特征、18S rRNA基因序列及系统进化树分析[J].宁波大学学报(理工版),2022,35(2):8-14.DOI:
ZHOU Ying,ZHU Bo,QIAN Dong,LU Xiandong,LI Xiaobing,LI Zheng,LIU Yanhong,CAI Huifeng,WU Zhongning,YUAN Siping,ZHANG Chonghu,YE Leping,MA Rongrong*.Pathological characteristics of Enterocytozoon hepatopenaei, 18S rRNA gene sequence and phylogenetic analysis[J].Journal of Ningbo University(Natural Science & Engineering Edition),2022,35(2):8-14.DOI:
| Title: | Pathological characteristics of Enterocytozoon hepatopenaei, 18S rRNA gene sequence and phylogenetic analysis |
| 作者: | 周 颖, 祝 波, 钱 冬, 卢先东, 李小冰, 李 政, 刘艳红, 蔡惠凤, 吴仲宁, 袁思平, 张崇虎, 叶乐平, 马荣荣* |
| Author(s): | ZHOU Ying, ZHU Bo, QIAN Dong, LU Xiandong, LI Xiaobing, LI Zheng, LIU Yanhong, CAI Huifeng, WU Zhongning, YUAN Siping, ZHANG Chonghu, YE Leping, MA Rongrong* |
| 关键词: | 虾肝肠胞虫; 凡纳对虾; 病理特征; 系统进化树 |
| Keywords: | 18S rRNA; Enterocytozoon hepatopenaei; Penaeus vannamei; pathological characteristics; phylogenetic tree |
| 分类号: | S945.1 |
| 文献标识码: | A |
| 摘要: | 为调查浙江省部分地区虾肝肠胞虫(Enterocytozoon hepatopenaei, EHP)暴发来源及与其他宿主来源的微孢子虫进化关系, 本研究利用形态学特征和分子鉴定, 尝试对引起浙江多地凡纳对虾生长缓慢综合征的病原进行探究. 同时基于18S rRNA基因序列分析该病原与其他地区及其他宿主来源微孢子虫的进化关系. 结果显示, 所采病虾均为EHP阳性, 采样地区EHP 18S rRNA基因具有高度同源性, 为98.5%~100%. 对国内外EHP基因序列进行同源性分析, 其同源性为86.2%~100%, 所采EHP与印度EHP聚为一个分支. EHP 18S rRNA基因序列与其他水生生物微孢子虫18S rRNA均处于不同分支, 表明18S rRNA可作为EHP的主要鉴别序列. 将本研究所采EHP与NCBI中同源性较高的比氏肠胞虫和金头鲷微孢子虫比较分析, 结果表明微孢子虫基因序列同源性与宿主进化发育程度有关, 宿主之间进化距离越近, 则微孢子虫基因序列同源性相聚越近. |
| Abstract: | Outbreaks of slow growth syndrome of Penaeus vannamei have occurred in some areas of Zhejiang Province. In the present study, pathological characteristics and molecular identification were determined to examine the pathogen causing slow growth syndrome of Penaeus vanamei in shrimp farms. The relationship was also investigated between this pathogen and the microsporidia from other regions and other host sources based on 18S rRNA gene sequence. The results show that all the shrimps with the disease were EHP positive, and the EHP 18S rRNA gene in the sampling area was high homology (98.5%-100%). The homology analysis of EHP gene sequences at home and abroad was 86.2%-100%. The collected EHP and Indian EHP were clustered into one branch. The sequence of EHP 18S rRNA gene was in different branches from that of other aquatic organisms, indicating that 18S rRNA could be used as the main identification sequence of EHP. The comparative analysis of EHP with Enterocytozoon bieneusi and Enterospora nucleophile showed that the gene sequence homology of microsporidium was related to the degree of host evolution and development. The closer the evolutionary distance between hosts, the closer the gene sequence homology of microsporidium. |
| 参考文献 /References: | [1] Stentiford G D, Feist S W, Stone D M, et al. Microsporidia: Diverse, dynamic, and emergent pathogens in aquatic systems[J]. Trends in Parasitology, 2013, 29(11):567-578. [2] Anderson I G, Shariff M, Nash G. A hepatopancreatic microsporidian in pond-reared tiger shrimp, Penaeus monodon, from Malaysia[J]. Journal of Invertebrate Pathology, 1989, 53(2):278-280. [3] Tourtip S, Wongtripop S, Stentiford G D, et al. Enterocytozoon hepatopenaei sp. nov. (Microsporida: Enterocytozoonidae), a parasite of the black tiger shrimp Penaeus monodon (Decapoda: Penaeidae): Fine structure and phylogenetic relationships[J]. Journal of Invertebrate Pathology, 2009, 102(1):21-29. [4] Kasthuri Raju K. Haematological parameters as predictive indicators of stress induced mortality in Pacific White Shrimp Penaeus vannamei (Boone, 1931) broodstock during transboundary shipment[J]. Indian Journal of Geo- Marine Sciences, 2017, 46(7):1440-1446. [5] 胡吉卉, 李正民, 段健诚, 等. 虾类肝肠胞虫流行病学研究进展[J]. 水产养殖, 2020, 41(11):1-4; 10. [6] Aranguren L F, Han J E, Tang K F J. Enterocytozoon hepatopenaei (EHP) is a risk factor for acute hepatopancreatic necrosis disease (AHPND) and septic hepatopancreatic necrosis (SHPN) in the Pacific white shrimp Penaeus vannamei[J]. Aquaculture, 2017, 471:37- 42. [7] Aranguren Caro L F, Mai H N, Pichardo O, et al. Evidences supporting Enterocytozoon hepatopenaei association with white feces syndrome in farmed Penaeus vannamei in Venezuela and Indonesia[J]. Diseases of Aquatic Organisms, 2020, 141:71-78. [8] Zhu B, Lu X D, Liu Y H, et al. Effects of Enterocytozoon hepatopenaei single-infection or co-infection with Vibrio parahaemolyticus on the hepatopancreas of Penaeus vannamei[J]. Aquaculture, 2022, 549:737726. [9] 农业农村部渔业渔政管理局, 全国水产技术推广总站, 中国水产学会. 中国渔业统计年鉴-2020[M]. 北京:中国农业出版社, 2020. [10] Goldner J. A modification of the Masson trichrome technique for routine laboratory purposes[J]. The American Journal of Pathology, 1938, 14(2):237-243. [11] 常晓晴. 虾肝肠胞虫病理学检测技术的建立[D]. 上海:上海海洋大学, 2018. [12] Wang L J, Lv Q, He Y T, et al. Integrated qPCR and staining methods for detection and quantification of Enterocytozoon hepatopenaei in shrimp Litopenaeus vannamei[J]. Microorganisms, 2020, 8(9):1366. [13] Ding Z F, Chen J Q, Lin J, et al. Development of in situ hybridization and real-time PCR assays for the detection of Hepatospora eriocheir, a microsporidian pathogen in the Chinese mitten crab Eriocheir sinensis[J]. Journal of Fish Diseases, 2017, 40(7):919-927. [14] Ou Y L, Jiang W, Roellig D M, et al. Characterizations of Enterocytozoon bieneusi at new genetic loci reveal a lack of strict host specificity among common genotypes and the existence of a canine-adapted Enterocytozoon species [J]. International Journal for Parasitology, 2021, 51(2/3): 215-223. [15] Palenzuela O, Redondo M J, Cali A, et al. A new intranuclear microsporidium, Enterospora nucleophila n. sp., causing an emaciative syndrome in a piscine host (Sparus aurata), prompts the redescription of the family Enterocytozoonidae[J]. International Journal for Parasitology, 2014, 44(3/4):189-203. [16] 常晓晴, 王元, 英娜, 等. 8种染色方法对组织切片中虾肝肠胞虫染色效果的比较[J]. 海洋渔业, 2019, 41(1): 91-99. [17] 王元. 脊尾白虾与三疣梭子蟹微孢子虫病的病原和病理[D]. 上海: 上海海洋大学, 2013. [18] Wang Y, Li X C, Fu G H, et al. Morphology and phylogeny of Ameson portunus n. sp. (Microsporidia) infecting the swimming crab Portunus trituberculatus from China[J]. European Journal of Protistology, 2017, 61(Pt A):122-136. [19] Small H J, Stentiford G D, Behringer D C, et al. Characterization of microsporidian Ameson herrnkindi sp. nov. infecting Caribbean spiny lobsters Panulirus argus [J]. Diseases of Aquatic Organisms, 2019, 136(3):209- 218. [20] Small H J, Meyer G R, Stentiford G D, et al. Ameson metacarcini sp. nov. (Microsporidia) infecting the muscles of dungeness crabs Metacarcinus magister from British Columbia, Canada[J]. Diseases of Aquatic Organisms, 2014, 110(3):213-225. [21] Liu X H, Wen M Q, Zhao Y L, et al. Morphological and molecular characterization of a new freshwater micro- sporidium, Jirovecia sinensis sp. n. (Microsporidia) infecting the coelomocytes of Branchiura sowerbyi (Oligochaeta: Naididae) in China[J]. Journal of Invertebrate Pathology, 2020, 173:107368. [22] Sokolova Y Y, Overstreet R M. A new microsporidium, Apotaspora heleios n. g., n. sp., from the Riverine grass shrimp Palaemonetes paludosus (Decapoda: Caridea: Palaemonidae)[J]. Journal of Invertebrate Pathology, 2018, 157:125-135. [23] Abdel-Baki A A S, Tamihi A F, Al-Qahtani H A, et al. Glugea jazanensis sp. nov. infecting Lutjanus bohar in the Red Sea: Ultrastructure and phylogeny[J]. Diseases of Aquatic Organisms, 2015, 116(3):185-190. [24] Azevedo C, Abdel-Baki A A S, Rocha S, et al. Ultrastructure and phylogeny of Glugea arabica n. sp. (Microsporidia), infecting the marine fish Epinephelus polyphekadion from the Red Sea[J]. European Journal of Protistology, 2016, 52:11-21. |
| 备注/Memo: | 收稿日期: 2021－09－13. 宁波大学学报（理工版）网址: http://journallg.nbu.edu.cn/ 基金项目: 宁波市公益性计划（202002N3062）; 三门县农业农村局项目（三招采-2019-GK013号）. 宁波大学学报（理工版）网址：http://journallg.nbu.edu.cn/第一作者: 周颖（1999－）, 女, 陕西铜川人, 在读硕士研究生, 主要研究方向: 水生动物医学. E-mail: zy18700697409@163.com *通信作者: 马荣荣（1990－）, 女, 山东菏泽人, 助理研究员, 主要研究方向: 水生动物医学. E-mail: marongrong@nbu.edu.cn |
