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《山东大学学报(理学版)》 ›› 2020, Vol. 55 ›› Issue (9): 102-110.doi: 10.6040/j.issn.1671-9352.0.2019.834

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鳗弧菌和副溶血弧菌对短蛸感染的病理学研究

冯艳微2,李赞2,王卫军2,孙国华2,徐晓辉2,杨建敏2,*()   

  1. 1. 上海海洋大学水产与生命学院，上海 201306
    2. 鲁东大学农学院，山东 烟台 264025
    3. 上海海洋大学上海水产养殖工程技术研究中心，上海 201306
  • 收稿日期:2019-11-25 出版日期:2020-09-20 发布日期:2020-09-17
  • 通讯作者: 杨建敏 E-mail:up@126.com
  • 作者简介:王金龙(1994—),男,硕士研究生,研究方向为贝类遗传育种. E-mail:wjl07011811@163.com
  • 基金资助:
    国家贝类产业技术体系基金资助项目(CARS-49);山东省自然科学基金资助项目(ZR2019BC052)

Pathological study on infection of Vibrio anguillarum and Vibrio parahemolyticus to Octopus ocellatus

Yan-wei FENG2,Zan LI2,Wei-jun WANG2,Guo-hua SUN2,Guo-hua SUN2,Xiao-hui XU2,*()   

  1. 1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
    2. Agricultural College, Ludong University, Yantai 264025, Shandong, China
    3. Shanghai Aquaculture Engineering Technology Research Center, Shanghai Ocean University, Shanghai 201306, China
  • Received:2019-11-25 Online:2020-09-20 Published:2020-09-17
  • Contact: Xiao-hui XU E-mail:up@126.com

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摘要：

为了今后养殖病害的防治,了解病原微生物在养殖过程中对头足类动物产生的影响,选择在海水养殖中发病频率较高的弧菌,包括鳗弧菌(Vibrio anguillarum)和副溶血弧菌(Vibrio Parahemolyticus),研究它们对短蛸的毒力水平。通过将稀释菌液直接注射短蛸腕部的方式,观察感染后短蛸的致死率、半致死时间,以及各菌对短蛸肝脏、鳃、鳃心、白体等器官结构的影响。结果显示:鳗弧菌和副溶血弧菌均对短蛸有较强的致病性,分别在60、48 h时达到半致死,且发病的短蛸活性减退,虚弱无力,腹部肿大,解剖可见部分内脏肿胀明显,如肝脏;通过对病变组织和健康组织的石蜡切片和HE染色,以及超显微结构的观察,发现患病短蛸的肝脏、鳃、鳃心和白体均出现不同程度组织、细胞结构上的破坏。以上结果表明这两种弧菌对短蛸均具有明显的致病性,副溶血弧菌更强一些,合适条件下将会成为短蛸规模化养殖的重要威胁之一。

关键词: 短蛸, 鳗弧菌, 副溶血弧菌, 感染

Abstract:

In order to control aquaculture diseases and understand the influence of pathogen microorganisms on cephalopoda culture, we selected vibrio with high incidence in seawater culture, including V.anguillarum and V.parahemolyticus, to study their virulence level and pathogenic mechanism against shortarm octopus. Through direct injection of diluted bacteria solution into shortarm octopus carpus arm, the death rate and half lethal time of shortarm octopus after infection were observed, as well as the effects of various bacteria on the structures of shortarm octopus liver, gill, gill heart and white body. The results showed that V.anguillarum and V.parahemolyticus were both highly pathogenic to shortarm octopus, reaching a semi-fatality rate at 60 h and 48 h, respectively. In addition, the activity of octopus was decreased, weakness and abdominal swelling were observed; Through the observation of paraffin sections and HE staining and ultramicroscopic structure of diseased and healthy tissues, it was found that the liver, gill, gill heart and white body of the diseased octopus showed different degrees of tissue or cell structure damage. The above results indicated that both V.anguillarum and V.parahemolyticus were significantly pathogenic to shortarm octopus, and the pathogenicity of V.parahaemolyticus might be stronger, which would be one of the greatest threats of shortarm octopus large-scale farming if the conditions were suitable.

Key words: Octopus ocellatus, Vibrio anguillarum, Vibrio Parahemolyticus, infection

中图分类号: 

  • Q17

表1

短蛸感染弧菌后各时间段的存活数变化"

组别时间/h
0 12 24 36 48 60 72 96 120
对照组(C)/只 16 16 16 15 15 15 15 15 15
鳗弧菌感染组(A)/只 16 16 15 14 11 8 6 2 0
副溶血弧菌感染组(B)/只 16 16 13 10 8 6 3 1 0

图1

短蛸肝脏和鳃组织的石蜡切片和HE染色(A、B分别代表肝脏和鳃组织)"

图2

短蛸鳃心和白体组织的石蜡切片和HE染色(C、D分别代表鳃心和白体组织, 0—2分别代表对照组、鳗弧菌感染组、副溶血弧菌感染组)"

图3

短蛸肝脏和鳃组织感染后的透射电镜分析(E、F分别代表肝脏和鳃组织, 0—2分别代表对照组、鳗弧菌感染组、副溶血弧菌感染组, N代表细胞核, CP代表细胞质,黑箭头指向鳃部分泌细胞,白箭头指向鳃上皮微绒毛结构)"

图4

短蛸鳃心和白体组织感染后的透射电镜分析(G、H分别代表鳃心和白体, 0—2分别代表对照组、鳗弧菌感染组、副溶血弧菌感染组, N代表细胞核, CP代表细质, DB代表致密体结构)"

1 董正之. 中国动物志:软体动物门头足纲[M]. 北京: 科学出版社, 1988: 174- 176.
DONG Zhengzhi . Zoology of China: Mollusca Cephalopoda[M]. Beijing: Science Press, 1988: 174- 176.
2 张伟伟, 雷晓凌. 短蛸不同组织的营养成分分析与评价[J]. 湛江海洋大学学报, 2006, 26 (4): 91- 93.
ZHANG Weiwei , LEI Xiaoling . Analysis and evaluation of nutritional components in different tissues of Octopus amulet[J]. Journal of Zhanjiang Ocean University, 2006, 26 (4): 91- 93.
3 马之明, 徐实怀, 贾晓平. 蛸类渔业概况及增养殖研究现状与展望[J]. 南方水产科学, 2008, 4 (5): 69- 73.
MA Zhiming , XU Shihuai , JIA Xiaoping . Research status and exploitation prospect in production basic bioligy aquaculture of Octopus[J]. South China Fisheries Science, 2008, 4 (5): 69- 73.
4 吕国敏, 吴进锋, 陈利雄. 我国头足类增养殖研究现状及开发前景[J]. 南方水产科学, 2007, 3 (3): 61- 66.
LYU Guoming , WU Jinfeng , CHEN Lixiong . Research achievements and exploitation prospect of Cephalopoda aquaculture in China[J]. South China Fisheries Science, 2007, 3 (3): 61- 66.
5 徐实怀, 马之明, 贾晓平. 人工养殖条件下真蛸的生物学特性及胚胎发育[J]. 南方水产科学, 2009, 5 (2): 63- 68.
XU Shihuai , MA Zhiming , JIA Xiaoping . Biological characteristics embryonic development of Octopus vulgaris under artificial farming condition[J]. South China Fisheries Science, 2009, 5 (2): 63- 68.
6 宋坚, 肖登兵, 宫海宁. 长蛸生物学及增养殖研究进展[J]. 河北渔业, 2013, (11): 55- 57.
SONG Jian , XIAO Dengbing , GONG Haining . Advances in biology and breeding of Octopus amulet[J]. Hebei Fisheries, 2013, (11): 55- 57.
7 金岳, 陈新军. 中国近海头足类基础生物学研究进展[J]. 海洋渔业, 2017, 39 (6): 696- 712.
JIN Yue , CHEN Xinjun . Advances in basic biology of Cephalopods in offshore China[J]. Marine Fisheries, 2017, 39 (6): 696- 712.
8 郑小东, 王如才, 王昭萍. 头足类遗传变异研究进展[J]. 水产学报, 2001, 25 (1): 84- 89.
ZHENG Xiaodong , WANG Rucai , WANG Shaoping . Advance of studies on Cephalopoda genetic variation[J]. Journal of Fisheries of China, 2001, 25 (1): 84- 89.
9 HANLON R T , FORSYTHE J W , COOPER K M , et al. Fatal penetrating skin ulcers in laboratory-reared Octopuses[J]. Journal of Invertebrate Pathology, 1984, 44 (1): 67- 83.
doi: 10.1016/0022-2011(84)90047-8
10 FORSYTHE J W . Octopus joubini (Mollusca: Cephalopoda): a detailed study of growth through the full life cycle in a closed seawater system[J]. Proceedings of the Zoological Society of London, 1984, 202 (3): 393- 417.
doi: 10.1111/j.1469-7998.1984.tb05091.x
11 EGIDIUS E . Vibriosis: pathogenicity and pathology. a review[J]. Aquaculture, 1987, 37 (1/2): 15- 28.
12 方伟, 杨杏芬, 柯昌文. 副溶血性弧菌分型研究进展[J]. 中华疾病控制杂志, 2008, 12 (5): 468- 472.
FANG Wei , YANG Xingfang , KE Changwen . Advances in the classification of Vibrio parahaemolyticus[J]. Chinese Journal of Disease Control, 2008, 12 (5): 468- 472.
13 王瑞, 于佳佳, 王鸿霞, 等. 副溶血弧菌攻毒过程中文蛤肝胰腺弧菌载量变化的分析[J]. 海洋科学, 2019, 43 (4): 29- 35.
WANG Rui , YU Jiajia , WANG Hongxia , et al. Vibrio load variation in the hepatopancreas of clam (Meretrix petechialis) under Vibrio parahaemolyticus challenge[J]. Marine Science, 2019, 43 (4): 29- 35.
14 林强, 李宁求, 付小哲, 等. 基于层次分析法的牡蛎养殖过程中副溶血弧菌风险评估模型建立和初步应用[J]. 海洋湖沼通报, 2017, (2): 116- 122.
LIN Qiang , LI Ningqiu , FU Xiaozhe , et al. Establishment and application of risk assessment model of Vibrio parahaemolyticus in oyster culture course through analytic hierarchy processing[J]. Transactions of Oceanology and Limnology, 2017, (2): 116- 122.
15 王小刚. 虾蟹肝胰腺组织学研究进展[J]. 广西水产科技, 2015, (1): 9- 15.
WANG Xiaogang . Progress in histological study of shrimps and crabs hepatopancreatic[J]. Guangxi Aquatic Science and Technology, 2015, (1): 9- 15.
16 邢婧. 栉孔扇贝稚贝造血组织的研究[J]. 水产学报, 2005, 29 (2): 145- 149.
XING Jing . Localization of hematopoietic tissue of scallop, Chamys farreri[J]. Journal of Fisheries of China, 2005, 29 (2): 145- 149.
17 蔡文超, 区又君. 驼背鲈肾脏显微和超微结构观察[J]. 中国水产科学, 2008, (5): 773- 778.
CAI Wenchao , OU Youjun . Microscopic and ultrastructural observation of the kidney of perch[J]. Journal of Fishery Sciences of China, 2008, (5): 773- 778.
18 NYHOLM S V , STEWART J J , RUBY E G , et al. Recognition between Symbiotic Vibrio Fischeri and the Haemocytes of Euprymna Scolopes[J]. Environmental Microbiology, 2009, 11 (2): 483- 493.
doi: 10.1111/j.1462-2920.2008.01788.x
19 李凌.栉孔扇贝抗鳗弧菌感染性状候选基因的多态性研究[D].青岛:中国科学院研究生院(海洋研究所), 2009.
LI Ling. The polymorphisms of candidate genes associated with susceptibility of Zhikong Scallop to Listonella anguillarum.[D]. Qingdao: Graduate School of the Chinese Academy of Sciences (Institute of Oceanography), 2009.
20 杨捷频. 常规石蜡切片方法的改良[J]. 生物学杂志, 2006, 23 (1): 45- 46.
YANG Jiepin . Improvement of conventional paraffin section method[J]. Journal of Biology, 2006, 23 (1): 45- 46.
21 金银姬, 李长玲, 曹伏君, 等. 红笛头肾和脾脏显微结构观察[J]. 台湾海峡, 2006, 25 (3): 360- 367.
JIN Yinji , LI Changling , CAO Fujun , et al. Microstructure of the kidney and spleen of the red flute head[J]. Journal of Oceanography Taiwan Strait, 2006, 25 (3): 360- 367.
22 PEDERSEN K , KVHN I , SEPPNEN J , et al. Clonality of Vibrio anguillarum strains isolated from fish from the Scandinavian countries, Sweden, Finland and Denmark[J]. Journal of Applied Microbiology, 1999, 86 (2): 337- 347.
doi: 10.1046/j.1365-2672.1999.00658.x
23 WESTERDAHL A , OLSSON J C , CONWAY P L , et al. Characterisation of turbot (Scophthalmus maximus) associated bacteria with inhibitory effect against the fish pathogen Vibrio anguillarum[J]. Acta Microbiologica Et Immunologica Hungarica, 1994, 41 (4): 403- 409.
24 夏永娟, 黄威权, 姜国良. 鳗弧菌感染牙鲆的组织学和免疫组织化学观察[J]. 海洋科学, 2000, 24 (10): 41- 44.
XIA Yongjuan , HUANG Weiquan , JIANG Guoliang . Histological and immunohistochemical observations of Japanese flounder infected by Vibrio anguillus[J]. Marine Science, 2000, 24 (10): 41- 44.
25 谢嘉华, 翁祖英, 袁建军. 两种病原菌感染后海鳗肝脏的组织病理学研究[J]. 泉州师范学院学报, 2008, 26 (4): 71- 74.
XIE Jiahua , WENG Zuying , YUAN Jianjun . Study on histopathology of liver of Pike Eel Muraenesox Cinereus infected with two species of fish bacterial pathogens[J]. Journal of Quanzhou Normal University, 2008, 26 (4): 71- 74.
26 张彬, 黄婷, 熊建华, 等. 文蛤主要弧菌性病害研究进展[J]. 广东农业科学, 2012, 39 (17): 128- 130.
ZHANG Bin , HUANG Ting , XIONG Jianhua , et al. Research progress of vibriosis in hand calm, Meretrix meretrix Linnaeus[J]. Guangdong Agricultural Science, 2012, 39 (17): 128- 130.
27 李天道, 于佳, 俞开康. 四种弧菌对中国对虾的致病性研究[J]. 海洋湖沼通报, 1998, (1): 57- 64.
LI Tiandao , YU Jia , YU Kaikang . Study on the pathogenicity of four Vibrio strains to Penaeus chinensis[J]. Transactions of Oceanology and Limnology, 1998, (1): 57- 64.
28 FISH ELSON L . Cytomorphological alterations of the thymus, spleen, head-kidney, and liver in cardinal fish (Apogonidae, Telestei)as bioindicators of stress[J]. J Morph, 2006, 267: 57- 69.
doi: 10.1002/jmor.10385
29 刘振勇, 谢友佺, 林小金, 等. 大黄鱼肝脏病变组织病理学观察[J]. 渔业科学进展, 2007, 28 (5): 7- 11.
LIU Zhenyong , XIE Youquan , LIN Xiaojin , et al. Histopathological observation of pathological changes in the liver of Pseudosciaena crocea[J]. Advances in Fishery Science, 2007, 28 (5): 7- 11.
30 鲁双庆, 刘和军, 刘红玉, 等. Cu2+或表面活性剂AE对黄鳝肝损伤的超微结构观察[J]. 应用与环境生物学报, 2003, 9 (2): 167- 170.
LU Shuangqing , LIU Hejun , LIU Yuhong , et al. Ultra-structural observation of Monopterus albus liver cells exposed to Cu2+ or surfantant AE pollution[J]. Chin J Appl Environ Biol, 2003, 9 (2): 167- 170.
31 冯涛, 魏凤琴, 欧阳高亮, 等. 苯并(a)芘对大弹涂鱼肝细胞超微结构的影响[J]. 应用生态学报, 2003, 14 (10): 1780- 1782.
FENG Tao , WEI Fengqin , OUYANG Liangliang , et al. Effect of benzo (a) pyrene on the ultrastructure of liver cells in bomb-coated fish[J]. Journal of Applied Ecology, 2003, 14 (10): 1780- 1782.
32 耿毅, 汪开毓, 叶仕根, 等. 鲤一种粘孢子虫病的病理学研究[J]. 水产科学, 2003, (1): 11- 13.
GENG Yi , WANG Kaiyu , YE Shigen , et al. A pathological study on myxosporidiosis of carp[J]. Fisheries Science, 2003, (1): 11- 13.
33 FISHELSON L , BECKER K . Development and aging of the liver and pancreas in the domestic carp, cyprinus carpio: from embryogenesis to 15-year-old Fish[J]. Environmental Biology of Fishes, 2001, 61 (1): 85- 97.
34 杜雄伟, 常藕琴, 王晓辉, 等. 嗜水气单胞菌对剑尾鱼的致病性及组织病理学研究[J]. 长江大学学报(自科版), 2005, 2 (2): 53- 56.
DU Xiongwei , CHANG Ouqin , WANG Xiaohui , et al. Study on the pathogenicity and histopathology of Aeromonas hydrophila to swordtail fish[J]. Journal of Yangtze University(Nat Sci Edi), 2005, 2 (2): 53- 56.
35 黄琪琰, 郑德崇, 金丽华, 等. 淡水鱼类细菌性败血症的组织亚显微病理及防治研究[J]. 上海海洋大学学报, 1995, 4 (1): 27- 34.
HUANG Qiyan , ZHENG Dezong , JIN Lihua , et al. Studies on the bacterial septicema of freshwater fishes: the super-microhisto pathology, prevention and cure[J]. Journal of Shanghai Fisheries University, 1995, 4 (1): 27- 34.
36 刘彦, 李健, 王群, 等. 鳗弧菌感染牙鲆药物代谢动力学模型的初步建立[J]. 渔业科学进展, 2007, 28 (2): 76- 83.
LIU Yan , LI Jian , WANG Qun , et al. Establishment of a pharmacokinetic model for Paralichthys olivaceus with Vibrio anguillarum[J]. Advances in Fishery Science, 2007, 28 (2): 76- 83.
37 HU M Y , PO-HSUAN S , YING-JEY G , et al. Perfused gills reveal fundamental principles of pH regulation and ammonia homeostasis in the Cephalopod Octopus vulgaris[J]. Frontiers in Physiology, 2017, (8)
doi: 10.3389/fphys.2017.00162
38 段亚飞, 梁俊平, 李吉涛, 等. 鳗弧菌感染对脊尾白虾鳃组织抗氧化酶系统的影响[J]. 水产科学, 2015, 34 (11): 667- 672.
DUAN Yafei , LIANG Junping , LI Jitao , et al. Antioxidant enzyme system in gill of shrimp carinicauda with Vibrio anguillarum[J]. Fisheries Science, 2015, 34 (11): 667- 672.
39 王朝辉, 尹伊伟. 珠海桂山岛米氏裸甲藻赤潮对鱼鳃损伤的病理学组织观察[J]. 海洋学报, 2001, 4 (1): 133- 137.
WANG Zhaohui , YIN Yiwei . Histopathological changes in fish gills during Cymnodinium mikimotoi red tide in Guishan island area, the South China Sea[J]. Acta Oceanologica Sinica, 2001, 4 (1): 133- 137.
40 王朝晖, 尹伊伟. 甲氰菊酯对白鲢(Hypophthalmichthys molitrix)鳃组织病理学研究[J]. 暨南大学学报(自然科学版), 1996, 17 (1): 74- 79.
WANG Zhaohui , YIN yiwei . Armor cyanogen chrysanthemum ester of silver carp (Hypophthalmichthys molitrix) gill tissue pathology research[J]. Journal of Jinan University(Natural Science), 1996, 17 (1): 74- 79.
41 中原元和, 陈舜华. 章鱼鳃心对钴-60的特殊积累[J]. 原子能农业译丛, 1980, (4): 38- 39.
ZHONGYUAN Yuanhe , CHEN Shunhua . Special accumulation of cobalt-60 in the gill heart of Octopus[J]. Atomic Energy Agriculture, 1980, (4): 38- 39.
42 BEUERLEIN K , LÖHR S , WESTERMANN B , et al. Components of the cellular defense and detoxification system of the common cuttlefish Sepia officinalis (Mollusca, Cephalopoda)[J]. Tissue and Cell, 2002, 34 (6): 390- 396.
doi: 10.1016/S0040816602000708
43 FIEDLER A , SCHIPP R . Localization of catecholamine-containing nerve fibres in the branchial heart and cardiac ganglion of the common cuttlefish Sepia officinalis L. (cephalopoda)[J]. Tissue and Cell, 1991, 23 (6): 813- 819.
doi: 10.1016/0040-8166(91)90033-P
44 SALAZAR K A , JOFFE N R , NATHALIE D , et al. Transcriptome analysis of the white body of the squid Euprymna tasmanica with emphasis on immune and hematopoietic gene discovery[J]. PLoS One, 2015, 10 (3)
45 JOHNSON E F . The vasculature of the white bodies in the ommastrephid squid Illex illecebrosus (LeSueur, 1821): a proposed route for the dissemination of newly formed hemocytes[J]. Canadian Journal of Zoology, 1987, 65 (7): 1607- 1620.
doi: 10.1139/z87-250
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[2] 杨永伟1,2,贺鹏飞2,李毅君2,3. BL-代数的严格滤子[J]. 山东大学学报（理学版）, 2014, 49(03): 63 -67 .
[3] 李敏1,2,李歧强1. 不确定奇异时滞系统的观测器型滑模控制器[J]. 山东大学学报（理学版）, 2014, 49(03): 37 -42 .
[4] 刘婷婷,陈志勇,李晓琴*,杨文志. 随机变量序列的Berry-Esseen界[J]. 山东大学学报（理学版）, 2014, 49(03): 101 -106 .
[5] 韩亚飞,伊文慧,王文波,王延平,王华田*. 基于高通量测序技术的连作杨树人工林土壤细菌多样性研究[J]. 山东大学学报（理学版）, 2014, 49(05): 1 -6 .
[6] 马媛媛, 孟慧丽, 徐久成, 朱玛. 基于粒计算的正态粒集下的格贴近度[J]. 山东大学学报（理学版）, 2014, 49(08): 107 -110 .
[7] 邱桃荣,王璐,熊树洁,白小明. 一种基于粒计算的知识隐藏方法[J]. J4, 2010, 45(7): 60 -64 .
[8] 薛秋芳1,2,高兴宝1*,刘晓光1. H-矩阵基于外推GaussSeidel迭代法的几个等价条件[J]. J4, 2013, 48(4): 65 -71 .
[9] 王 琦,赵秀恒,李国君 . 超图在树环中的嵌入问题[J]. J4, 2007, 42(10): 114 -117 .
[10] 史爱玲1,马明2*,郑莹2. 齐次泊松响应的客户寿命值及性质[J]. 山东大学学报（理学版）, 2014, 49(03): 96 -100 .
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