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

doi:10.3969/j.issn.1004-1524.2022.08.06

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (8): 1617-1625.DOI: 10.3969/j.issn.1004-1524.2022.08.06

• Animal Science • Previous Articles Next Articles

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

Abstract

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

CLC Number:

S947.2

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.

Figures/Tables 8

References 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

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

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