棘胸蛙脑膜炎败血伊丽莎白菌的分离鉴定及其感染的病理损伤

doi:10.3969/j.issn.1004-1524.2018.03.03

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (3): 371-377.DOI: 10.3969/j.issn.1004-1524.2018.03.03

棘胸蛙脑膜炎败血伊丽莎白菌的分离鉴定及其感染的病理损伤

雷雪平^a, 耿毅^a,*, 余泽辉^a, 郑李平^a, 曹师琪^a, 黄小丽^b, 陈德芳^b, 欧阳萍^a, 刘恺睿^a

a.四川农业大学动物医学院,四川成都 611134;
b.四川农业大学水产系,四川成都 611134

收稿日期:2017-11-27 出版日期:2018-03-20 发布日期:2018-03-21
通讯作者: *耿毅,E-mail:gengyisicau@126.com
作者简介:雷雪平(1993—),女,四川绵阳人,硕士,研究方向为水生动物病原学与病理学。E-mail: leixpingsicau @163.com
基金资助:
四川省淡水鱼产业技术体系创新团队建设项目(2017SICAD002);四川省重点研发项目(182DYF)

Isolation and identification of Elizabethkingia meningoseptica from Chinese spiny frog (Quasipaa spinosa) and pathological lesions of its infection

LEI Xueping^a, GENG Yi^a,*, YU Zehui^a, ZHENG Liping^a, CAO Shiqi^a, HUANG Xiaoli^b, CHEN Defang^b, OUYANG Ping^a, LIU Kairui^a

a.College of Veterinary Medicine Sichuan Agricultural University, Chengdu 611134, China;
b.Department of Aquaculture, Sichuan Agricultural University, Chengdu 611134, China

Received:2017-11-27 Online:2018-03-20 Published:2018-03-21

摘要/Abstract

摘要： 2016年4月四川雅安一棘胸蛙(Quasipaa spinosa)养殖场暴发一种临床特征表现为白内障、体表溃疡和神经症状的传染病。为明确此次流行病病因,本研究进行了病原菌分离、人工感染、分离菌理化特性与16S rDNA基因序列分析。结果从患病蛙肝、脾、肾中分离到一株G^-短杆菌(CM160701);将其腹腔注射感染健康棘胸蛙表现出与自然发病蛙相似症状。该菌的LD₅₀为1.19×10⁶ cfu·只^-1;通过理化特性与16S rDNA基因序列分析鉴定分离菌为脑膜炎败血伊丽莎白菌(Elizabethkingia meningoseptica);其对氟苯尼考敏感,对恩诺沙星、多西环素、阿莫西林等多种药物耐受。病理组织学观察发现,其感染对全身多组织器官造成损伤,表现出明显的变性、坏死和炎症反应,尤以脑、肝、脾、心和肾损伤严重。该研究证实了脑膜炎败血伊丽莎白菌是本次疫情的病因,其感染引起棘胸蛙全身性损伤导致多组织器官功能障碍而死亡。

关键词: 棘胸蛙, 脑膜炎败血伊丽莎白菌, 生物学特性, 病理损伤

Abstract: In April 2016, a serious infectious disease characterized by cataract, cutaneous ulcer and neurological symptoms emerged in a Quasipaa spinosa farm in Ya'an, Sichuan Province. In order to explore the etiology, the pathogen isolation, artificial infection, physical and chemical characteristics of isolated bacteria and sequence analysis of 16S rDNA gene were analyzed. The results showed that a Gram-negative, short rod-shaped bacillus (CM160701) was isolated from the diseased frogs. Healthy Chinese spiny frogs injected the isolate intraperitoneally exhibited the similar clinical signs, gross and histopathological lesions to the naturally infected frogs, and the LD₅₀ value of the isolate was 1.19×10⁶ cfu per frog. The isolated strain was identified to be Elizabethkingia meningoseptica according to physicochemical properties and the 16S rDNA gene sequence analysis. The strain was sensitive to florfenicol, but resistant to enrofloxacin, amoxicillin, doxycycline and so on. Histopathological observation found that the E. meningoseptica infection in Quasipaa spinosa could cause pathological damages to multiple organs which showed degeneration, necrosis and inflammation significantly, especially in the brain, liver, spleen, heart and kidney. This study confirmed E. meningoseptica was the pathogen of the epidemic, and the E. meningoseptica infection could cause Quasipaa spinosa multi-system injuries and lead to death.

Key words: Quasipaa spinosa, Elizabethkingia meningoseptica, biological characteristics, pathological lesions

中图分类号:

S947.2

雷雪平, 耿毅, 余泽辉, 郑李平, 曹师琪, 黄小丽, 陈德芳, 欧阳萍, 刘恺睿. 棘胸蛙脑膜炎败血伊丽莎白菌的分离鉴定及其感染的病理损伤[J]. 浙江农业学报, 2018, 30(3): 371-377.

LEI Xueping, GENG Yi, YU Zehui, ZHENG Liping, CAO Shiqi, HUANG Xiaoli, CHEN Defang, OUYANG Ping, LIU Kairui. Isolation and identification of Elizabethkingia meningoseptica from Chinese spiny frog (Quasipaa spinosa) and pathological lesions of its infection[J]. , 2018, 30(3): 371-377.

参考文献

[1] KIM K K, KIM M K, LIM J H, et al.Transfer of Chryseobacterium meningosepticum and Chryseobacterium miricola to Elizabethkingia gen. nov. as Elizabethkingia meningoseptica comb. nov. and Elizabethkingia miricola comb. nov.[J]. International Journal of Systematic & Evolutionary Microbiology, 2005, 55(Pt 3):1287.
[2] STEINBERG J P, BURD E M. 238 - Other Gram-negative and Gram-variable Bacilli[J]. Mandell Douglas & Bennetts Principles & Practice of Infectious Diseases, 2015, 2:2667-2683.
[3] URSING J, BRUUN B.Genetic heterogeneity of Flavobacterium meningosepticum demonstrated by DNA-DNA hybridization[J]. Acta Pathologica Microbiologica Et Immunologica Scandinavica, 1987, 95(1):33-39.
[4] JACOBS A, CHENIA H Y.Biofilm formation and adherence characteristics of an Elizabethkingia meningoseptica isolate from Oreochromis mossambicus[J]. Annals of clinical microbiology and antimicrobials, 2011, 10:16.
[5] JEAN S S, LEE W S, CHEN F L, et al.Elizabethkingia meningoseptica: an important emerging pathogen causing healthcare-associated infections[J]. Journal of Hospital Infection, 2014, 86(4):244-249.
[6] VANCANNEYT M, SEGERS P, HAUBEN L, et al.Flavobacterium meningosepticum, a pathogen in birds[J]. Journal of Clinical Microbiology, 1994, 32(10):2398-2403.
[7] DANNA Y U, ZHENG R Q, QIN F, et al.Genetic diversity and population structure for the conservation of giant spiny frog (Quasipaa spinosa) using microsatellite loci and mitochondrial DNA[J]. Asian Herpetological Research, 2016, 7(2):75-86.
[8] 梅祎芸,叶容晖, 宋婷婷, 等. 浙江省棘胸蛙养殖现状及发展对策[J]. 浙江农业科学,2015, 1(7):1122-1125.
MEI Y Y, YE R H, SONG T T, et al.Current situation and Development Countermeasures of Rana spinosa breeding in Zhejiang[J]. Journal of Zhejiang Agricultural Sciences, 2015, 1(7):1122-1125. (in Chinese)
[9] 陶吉兴,刘安兴,孙孟军. 浙江重点两栖动物种群数量研究[J]. 浙江大学学报(农业与生命科学版), 2004, 30(5):536-540.
TAO J X, LIU A X, SUN M J.Research on population quantity of main amphibian in Zhejiang province[J]. Journal of Zhejiang University (Agriculture and Life Sciences), 2004, 30(5):536-540. (in Chinese with English abstract)
[10] 叶容晖. 棘胸蛙微卫星分子标记筛选及其种群遗传分析[D]. 金华:浙江师范大学, 2009.
YE R H.SSR isolation and population genetic of Paa spinosa [D]. Jinhua: Zhejiang Normal University, 2009. (in Chinese with English abstract)
[11] JENSEN S, BERGH O, ENGER O, et al.Use of PCR-RFLP for genotyping 16S rRNA and characterizing bacteria cultured from halibut fry[J]. Canadian Journal of Microbiology, 2002, 48(5):379-386.
[12] 余泽辉,张佳,耿毅, 等. 齐口裂腹鱼无乳链球菌的分离鉴定及其感染的病理损伤[J]. 中国水产科学. 2014, 21(6):1244-1252.
YU Z H, ZHANG J, GENG Y, et al.Isolation, identification and pathogenicity of Streptococcus agalactiae from Schizothorax prenanti[J]. Journal of Fishery Sciences of China, 2014, 21(6):1244-1252. (in Chinese with English abstract)
[13] HOWARD A M, PAPICH M G, FELT S A, et al.The pharmacokinetics of enrofloxacin in adult African clawed frogs (Xenopus laevis)[J]. Journal of the American Association for Laboratory Animal Science, 2010, 49(6):800-804.
[14] GREEN S L, BOULEY D M, TOLWANI R J, et al.Identification and management of an outbreak of Flavobacterium meningosepticum infection in a colony of South African clawed frogs (Xenopus laevis)[J]. Journal of the American Veterinary Medical Association, 1999, 214(12):1792-1793.
[15] RANSANGAN J, ZAINURI N, LAL T M, et al.Identification of Elizabethkingia meningoseptica from American bullfrog (Rana catesbeiana) farmed in Sabah, Malaysia using PCR method and future management of outbreak[J]. Malaysian Journal of Microbiology, 2013, 9(1):769-776.
[16] MAUEL M J, MILLER D L, FRAZIER K S, et al.Bacterial pathogens isolated from cultured bullfrogs (Rana castesbeiana)[J]. Journal of Veterinary Diagnostic Investigation, 2002, 14(5):431-433.
[17] TAYLOR F R, SIMMONDS R C, LOEFFLER D G.Isolation of Flavobacterium meningosepticum in a colony of leopard frogs (Rana pipiens)[J]. Laboratory Animal Science, 1993, 43(1):105.
[18] XIE Z Y, ZHOU Y C, WANG S F, et al.First isolation and identification of Elizabethkingia meningoseptica from cultured tiger frog, Rana tigerina rugulosa[J]. Veterinary Microbiology, 2009, 138(1/2):140-144.
[19] DASZAK P, CUNNINGHAM A A, Hyatt A D.Infectious disease and amphibian population declines[J]. Diversity & Distributions, 2003, 9(2):141-150.
[20] ROLLINS-SMITH L A. Amphibian immunity-stress, disease, and climate change[J]. Developmental & Comparative Immunology, 2017, 66:111-119.
[21] BELLAIS S, LEOTARD S, POIREL L, et al.Molecular characterization of a carbapenem-hydrolyzing β-lactamase from Chryseobacterium(Flavobacterium) indologenes[J]. FEMS Microbiology Letters, 1999, 171(2):127-132.
[22] YUM J H, LEE E Y, HUR S H, et al.Genetic diversity of chromosomal metallo-beta-lactamase genes in clinical isolates of Elizabethkingia meningoseptica from Korea[J]. Journal of Microbiology, 2010, 48(3):358-364.
[23] 李明,宋婷婷,郑荣泉,等. 棘胸蛙歪头病病原分离·鉴定与药敏试验[J]. 安徽农业科学,2016,44(3):64-66.
LI M, SONG T T, ZHENG R Q, et al.Isolation, identification and drug sensitivity test of the pathogen of Quasipaa spinosa[J]. Journal of Anhui Agricultural Sciences, 2016, 44(3):64-66. (in Chinese with English abstract)

棘胸蛙脑膜炎败血伊丽莎白菌的分离鉴定及其感染的病理损伤

Isolation and identification of Elizabethkingia meningoseptica from Chinese spiny frog (Quasipaa spinosa) and pathological lesions of its infection

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	李戌清, 严建立, 阮松林. 三叶青炭疽病病原菌的鉴定与生物学特性[J]. 浙江农业学报, 2020, 32(11): 2009-2019.
[2]	李戌清, 张敬泽, 张雅, 吴根良. 浙江省绍兴市茄子黄萎病菌菌株致病型鉴定及其生物学特性研究[J]. 浙江农业学报, 2019, 31(5): 784-789.
[3]	康慧敏, 何嘉乐, 刘晨恺, 周瑶佳, 耿毅, 欧阳萍. 四川地区养殖锦鲤浮肿病毒检测与系统进化分析[J]. 浙江农业学报, 2019, 31(4): 539-544.
[4]	白明焕, 耿毅, 邓龙君, 甘维熊, 周剑, 黄小丽, 陈德芳, 欧阳萍, 赵若璇, 谌冰洁. 长薄鳅恶臭假单胞菌的分离鉴定及其感染的病理损伤[J]. 浙江农业学报, 2019, 31(2): 207-215.
[5]	周会明, 张焱珍, 柴红梅, 杨荣情, 谭莹, 张萍萍, 白玉英, 赵一莲, 金玉洁. 一株临沧野生黄鸡?的鉴定与生长条件研究[J]. 浙江农业学报, 2019, 31(10): 1655-1662.
[6]	马晓平, 杨秋霞, 俞演, 李德生, 王承东, 凌珊珊, 古玉. 大熊猫源枝孢样枝孢霉野生株(Z20)与突变株(Zt)部分生物学特性及药敏试验比较[J]. 浙江农业学报, 2018, 30(8): 1328-1335.
[7]	郑李平, 耿毅, 雷雪平, 余泽辉, 黄小丽, 陈德芳, 欧阳萍, 曹师琪, 韩锐. 鲟海豚链球菌的分离鉴定及其感染后的病理损伤[J]. 浙江农业学报, 2018, 30(2): 203-210.
[8]	刘子明, 金晶, 胡则辉, 戴庆敏, 李巧玲, 施倩, 吕耀平. 棘胸蛙白内障病原鉴定及病理组织观察[J]. 浙江农业学报, 2018, 30(11): 1811-1818.
[9]	马晓平, 杨天意, 俞演, 张志和, 王承东, 古玉. 大熊猫阴道源乳酸杆菌生物学特性研究[J]. 浙江农业学报, 2017, 29(7): 1093-1102.
[10]	谢放，张育，朱玉兰. 一株冬虫夏草相关菌株的生物学特性[J]. 浙江农业学报, 2016, 28(2): 306-.
[11]	李静静，丁松爽，李艳平，云菲，阎海涛，王志萌，刘国顺*. 生物炭与氮肥配施对烤烟干物质积累及土壤生物学特性的影响[J]. 浙江农业学报, 2016, 28(1): 96-.
[12]	周小军1,*，何晓婵1，郑寨生1，张尚法1，孙裕建1，楼兵干2. 莲藕腐败病病原菌生物学特性研究[J]. 浙江农业学报, 2015, 27(2): 211-.
[13]	王文杰，关伟军，马月辉，李向臣. 鸭胚脐带间充质干细胞的分离培养及其生物学特性[J]. 浙江农业学报, 2015, 27(11): 1903-.
[14]	李戌清1，田忠玲2，郑积荣1，郑经武2. 番茄灰叶斑病病原菌生物学特性及杀菌剂筛选[J]. 浙江农业学报, 2015, 27(11): 1953-.
[15]	王道泽;洪文英;吴燕君;柯汉云. 铁皮石斛害虫独角仙的生物学特性及防治技术研究[J]. , 2014, 26(3): 0-722729.