蛙类歪头、破头与白眼综合征病原分析

doi:10.3969/j.issn.1004-1524.2022.08.06

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (8): 1617-1625.DOI: 10.3969/j.issn.1004-1524.2022.08.06

蛙类歪头、破头与白眼综合征病原分析

李旭东¹(), 刘永涛², 杨先乐³, 杨移斌²^,^*(), 艾晓辉¹^,²

1.河南省水产技术推广站,河南郑州 450008
2.中国水产科学研究院长江水产研究所,湖北武汉 430223
3.上海海洋大学水产与生命学院,上海201306

收稿日期:2021-09-30 出版日期:2022-08-25 发布日期:2022-08-26
通讯作者: 杨移斌
作者简介:*杨移斌,E-mail: yangyb1988@126.com
李旭东(1980—),男,河南郑州人,硕士,工程师,主要从事水产动物病害研究。E-mail: 957918235@qq.com
基金资助:
现代农业产业技术体系专项资金(CARS-49);湖北省技术创新专项重大项目(2019AB077)

Analysis on pathogens of frogs with crooked head, broken head or white eye

LI Xudong¹(), LIU Yongtao², YANG Xianle³, YANG Yibin²^,^*(), AI Xiaohui¹^,²

1. Henan Fisheries Technology Promotion Station, Zhengzhou 450008, China
2. Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China
3. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China

Received:2021-09-30 Online:2022-08-25 Published:2022-08-26
Contact: YANG Yibin

摘要/Abstract

摘要：

为探明湖北、福建等主养区一系列蛙疾病的发病原因,从具有典型歪头、破头与白眼等症状的蛙脑部和内脏器官分离到40株优势菌。经16S rRNA 基因序列比对,发现40株优势菌均与米尔伊丽莎白菌(Elizabethkingia miricola)同源。经生理生化鉴定、系统发育分析发现,这40株优势菌存在遗传差异,宿主和区域是影响亲缘关系的重要因素,并判定菌株典型株QW08为米尔伊丽莎白菌。回归感染实验证实QW08为蛙类歪头、破头与白眼病的病原菌。组织病理学观察显示,米尔伊丽莎白菌感染黑斑蛙可引起其全身多脏器与组织损伤,脑、肠道、肝、脾与肾等组织器官损伤极其严重,主要病理变化表现为变性、坏死和炎症反应,致多组织器官功能障碍而引起感染蛙死亡。药敏特性分析显示,分离株QW08仅对氟苯尼考高度敏感,对其余19种抗生素表现出不同程度的耐药。研究结果为蛙类该病防控提供了理论依据。

关键词: 蛙, 米尔伊丽莎白菌, 分离鉴定, 病理, 药敏特性

Abstract:

In order to investigate the causes of a series of frog diseases in the main breeding areas of Hubei and Fujian, more than 40 strains of dominant bacteria were isolated from the brain and internal organs of frogs with typical symptoms of crooked head, broken head or white eyes. Through 16S rRNA gene sequence alignment, it was found that 40 dominant strains were homologous with Elizabethkingia miricola. After physiological and biochemical identification and phylogenetic analysis, the genetic differences were found in 40 dominant strains, the cluster analysis of 40 strains showed that host and region were important factors affecting genetic relationship, and the typical strain QW08 was identified as E. miricola. By regression infection experiment, strain QW08 was confirmed to be the pathogen of crooked head, broken head and white eye disease in frogs. Histopathological observations showed that E. miricola infection could cause damage to multiple organs and tissues in the whole body, including brain, intestinal tract, liver, spleen and kidney. The main pathological manifestations were degeneration, necrosis and inflammation, leading to dysfunction of multiple organs and tissues and death of infected frogs. Drug sensitivity analysis showed that QW08 was only highly sensitive to florfenicol, and showed different degrees of resistance to the other 19 antibiotics. The results provided a theoretical basis for the prevention and control of the disease in frogs.

Key words: frogs, Elizabethkingia miricola, isolation and identification, pathology, drug sensitivity

中图分类号:

S947.2

李旭东, 刘永涛, 杨先乐, 杨移斌, 艾晓辉. 蛙类歪头、破头与白眼综合征病原分析[J]. 浙江农业学报, 2022, 34(8): 1617-1625.

LI Xudong, LIU Yongtao, YANG Xianle, YANG Yibin, AI Xiaohui. Analysis on pathogens of frogs with crooked head, broken head or white eye[J]. Acta Agriculturae Zhejiangensis, 2022, 34(8): 1617-1625.

图/表 8

参考文献 25

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来源 Source	编号 Serial number	GenBank登录号 GenBank accession number
武汉黑斑蛙	QW08	MZ315056
Black spotted frog from Wuhan	QW09	MZ315057
	QW11	MZ315058
	QW12	MZ315059
	QW14	MZ315060
	QW15	MZ315061
荆州牛蛙	MW01	MZ315062
Bullfrog from Jingzhou	MW03	MZ315063
	MW04	MZ315064
	MW05	MZ315065
	MW06	MZ315066
	MW07	MZ315067
	MW10	MZ315068
	MW11	MZ315069
荆州黑斑蛙	QW02	MZ315070
Black spotted frog from Jingzhou	QW03	MZ315071
	QW04	MZ315072
	QW05	MZ315073
	QW06	MZ315074
	QW07	MZ315075
	QW10	MZ315076
	QW13	MZ315077
	QW16	MZ315078
	QW17	MZ315079
	QW18	MZ315080
漳州牛蛙	MW15	MZ315081
Bullfrog from Zhangzhou	MW16	MZ315082
	MW18	MZ315083
	MW19	MZ315084
	MW20	MZ315085
	MW12	MZ315086
	MW13	MZ315087
	MW22	MZ315088
	MW23	MZ315089
	MW24	MZ315090
	MW25	MZ315091
	MW26	MZ315092
	MW27	MZ315093
	MW28	MZ315094
	MW29	MZ315095

来源 Source	编号 Serial number	GenBank登录号 GenBank accession number
武汉黑斑蛙	QW08	MZ315056
Black spotted frog from Wuhan	QW09	MZ315057
	QW11	MZ315058
	QW12	MZ315059
	QW14	MZ315060
	QW15	MZ315061
荆州牛蛙	MW01	MZ315062
Bullfrog from Jingzhou	MW03	MZ315063
	MW04	MZ315064
	MW05	MZ315065
	MW06	MZ315066
	MW07	MZ315067
	MW10	MZ315068
	MW11	MZ315069
荆州黑斑蛙	QW02	MZ315070
Black spotted frog from Jingzhou	QW03	MZ315071
	QW04	MZ315072
	QW05	MZ315073
	QW06	MZ315074
	QW07	MZ315075
	QW10	MZ315076
	QW13	MZ315077
	QW16	MZ315078
	QW17	MZ315079
	QW18	MZ315080
漳州牛蛙	MW15	MZ315081
Bullfrog from Zhangzhou	MW16	MZ315082
	MW18	MZ315083
	MW19	MZ315084
	MW20	MZ315085
	MW12	MZ315086
	MW13	MZ315087
	MW22	MZ315088
	MW23	MZ315089
	MW24	MZ315090
	MW25	MZ315091
	MW26	MZ315092
	MW27	MZ315093
	MW28	MZ315094
	MW29	MZ315095

试验项目 Test items	QW08	米尔伊丽莎白菌 E. miricola
葡萄糖 Glucose	+	+
精氨酸双水解酶 Arginine dihydrolase	-	-
脲酶Urease	-	-
肌醇Inositol	-	-
柠檬酸盐Citrate	-	-
纤维二糖Cellobiose	-	-
水杨酸 Salicylic	-	-
H₂S产生 H₂S production	-	-
明胶Gelatin	+	+
氧化酶Oxidase	-	-
硝酸盐还原 Nitrate reduction	-	-
鸟氨酸脱羧酶 Ornithine decarboxyla	-	-
赖氨酸脱羧酶Lysin decarboxylase	-	-
乳糖Lactose	-	-
鼠李糖Rhamnose	-	-
蔗糖Saccharose	-	-
阿拉伯糖Arabinose	-	-
麦芽糖Maltose	-	-
棉子糖Raffinose	-	-
甘露糖 Mannose	+	+
吲哚 Indol	+	+
丙二酸盐 Malonate	-	-

试验项目 Test items	QW08	米尔伊丽莎白菌 E. miricola
葡萄糖 Glucose	+	+
精氨酸双水解酶 Arginine dihydrolase	-	-
脲酶Urease	-	-
肌醇Inositol	-	-
柠檬酸盐Citrate	-	-
纤维二糖Cellobiose	-	-
水杨酸 Salicylic	-	-
H₂S产生 H₂S production	-	-
明胶Gelatin	+	+
氧化酶Oxidase	-	-
硝酸盐还原 Nitrate reduction	-	-
鸟氨酸脱羧酶 Ornithine decarboxyla	-	-
赖氨酸脱羧酶Lysin decarboxylase	-	-
乳糖Lactose	-	-
鼠李糖Rhamnose	-	-
蔗糖Saccharose	-	-
阿拉伯糖Arabinose	-	-
麦芽糖Maltose	-	-
棉子糖Raffinose	-	-
甘露糖 Mannose	+	+
吲哚 Indol	+	+
丙二酸盐 Malonate	-	-

药物 Drug		抑菌圈直径判断标准 Judgment standard of inhibition zone diameter/mm			药物量 Dose/g	抑菌圈直径 Inhibition zone diameter/mm
药物 Drug		R	I	S	药物量 Dose/g	抑菌圈直径 Inhibition zone diameter/mm
β-内酰胺类β-Lactams	青霉素Penicillin	≤17	18~20	≥21	10	0^R
	阿莫西林Amoxicillin	≤13	14~17	≥18	20	0^R
头孢类Cephalosporin	头孢唑肟Ceftizoxime	≤14	15~19	≥20	30	18^I
	头孢拉定Cefradine	≤14	15~17	≥18	30	0^R
	头孢噻肟Cefotaxime	≤14	15~22	≥23	30	10^R
氨基糖苷类Aminoglycosides	庆大霉素Gentamicin	≤12	13~14	≥15	10	0^R
	链霉素Streptomycin	≤11	12~14	≥15	10	10^R
	奈替米星Netilmicin	≤12	13~14	≥15	30	0^R
	卡那霉素Kanamycin	≤13	14~17	≥18	30	0^R
	妥布霉素Tobramycin	≤12	13~14	≥15	10	0^R
	新霉素Neomycin	≤12	13~16	≥17	30	0^R
大环内酯类Macrolides	阿奇霉素Azithromycin	≤13	14~17	≥18	15	15^I
	红霉素Erythromycin	≤13	14~22	≥23	15	16^I
四环素类Tetracyclines	四环素Tetracycline	≤18	19~22	≥23	30	10^R
	强力霉素Doxycycline	≤12	13~15	≥16	30	12^R
喹诺酮类Quinolones	依诺沙星Enoxacin	≤14	15~17	≥18	10	12^R
	诺氟沙星Norfloxacin	≤12	13~16	≥17	10	8^R
酰胺醇类Amphenicols	氯霉素Chloramphenicol	≤12	13~17	≥18	300	15^I
	氟苯尼考Florfenicol	≤12	13~17	≥18	75	22^S
磺胺类Sulfonamides	磺胺异噁唑Sulfaisoxazole	≤12	13~16	≥17	300	0^R

蛙类歪头、破头与白眼综合征病原分析

Analysis on pathogens of frogs with crooked head, broken head or white eye

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