1株草龟维氏气单胞菌的分离鉴定及药敏谱分析

doi:10.11843/j.issn.0366-6964.2020.10.021

Abstract

Abstract: To determine the cause of death of Chinemys reevesii in a pet shop in Changli county, Hebei province, a dominant strain CLW-1 was isolated and purified from the diseased Chinemys reevesii. It was analyzed and identified through anatomical observation, bacterial isolation and culture, Gram staining and microscopic examination, biochemical identification and 16S rDNA gene sequencing, the drug resistance of the strain was detected through drug sensitivity test, and its pathogenicity was detected through zebrafish inoculation test and virulence gene experiment. The results showed that the bacteria formed moist, smooth surface, gray colony with β hemolysis on blood agar medium, and grew smooth and moist yellow colony on RS agar medium; Gram staining and microscopic examination showed negative coccidiosis; Homology analysis of 16S rDNA gene sequence showed that the isolated strain CLW-1 belongs to the same group with Aeromonas veronii, and the homology is above 99%; Inoculation test results of zebrafish showed that the strain had high pathogenicity, and its LD₅₀ was 1.26×10⁶ CFU; The virulence gene amplification results showed that the isolated strain CLW-1 had virulence genes of aerA, Exu, Lip and Ser; Drug sensitivity test results showed that the isolated strain CLW-1 was sensitive to piperacillin, cefazolin and enrofloxacin, etc; Susceptibility to furazolidone, oxacillin and erythromycin, etc; Resistance to midecamycin, clindamycin and vancomycin, etc. According to the test results, the isolated strain CLW-1 is confirmed to be Aeromonas veronii, which carries virulence genes and has high pathogenicity, it has great therapeutic effect by applying corresponding antibiotics, thus laying a foundation for the prevention and treatment of Aeromonas veronii.

Key words: Chinemys reevesii, Aeromonas veronii, isolation and identification, drug sensitivity test, virulence gene

CLC Number:

S852.61

WANG Miao, ZHANG Xuejia, DU Wannian, YU Xiujian, LIU Boxing, WU Tonglei, SHI Qiumei, ZHANG Zhiqiang. Isolation, Identification and Drug Sensitivity Test of a Strain of Aeromonas veronii from Chinemys reevesii[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(10): 2528-2535.

References 30

[1]	TIAN J X, KANG Y H, CHU G S, et al. Oral administration of Lactobacillus casei expressing flagellin a protein confers effective protection against aeromonas veronii in common carp, Cyprinus carpio[J]. Int J Mol Sci, 2020, 21(1):33.
[2]	VALÉRY M, BOURREL A S, GRIGNANO E, et al. Successful conservative treatment of necrotizing fasciitis associated with Aeromonas veronii in a patient with acute myeloid leukemia[J]. Med Mal Infect, 2020, doi:10. 1016/j. medmal. 2020. 01. 004.
[3]	李聪, 蔡岩, 周永灿, 等. 海南罗非鱼致病性维氏气单胞菌分离鉴定及药敏特性研究[J]. 水产科学, 2015, 34(10):640-646.LI C, CAI Y, ZHOU Y C, et al. Isolation, identification and antibiotic sensitivity of Aeromonas veronii from tilapia cultured in Hainan province[J]. Fisheries Science, 2015, 34(10):640-646. (in Chinese)
[4]	CHEN F, SUN J F, HAN Z R, et al. Isolation, identification and characteristics of aeromonas veronii from diseased crucian carp (Carassius auratus gibelio)[J]. Front Microbiol, 2019, 10:2742.
[5]	RAN C, QIN C B, XIE M X, et al. Aeromonas veronii and aerolysin are important for the pathogenesis of motile aeromonad septicemia in cyprinid fish[J]. Environ Microbiol, 2018, 20(9):3442-3456.
[6]	ZHANG D X, KANG Y H, ZHAN S, et al. Effect of Bacillus velezensis on Aeromonas veronii-induced intestinal mucosal barrier function damage and inflammation in crucian carp (Carassius auratus)[J]. Front Microbiol, 2019, 10:2663.
[7]	吴同垒, 单晓枫, 孟庆峰, 等. 维氏气单胞菌研究进展[J]. 中国兽药杂志, 2011, 45(7):41-44.WU T L, SHAN X F, MENG Q F, et al. Advances in Aeromonas veronii[J]. Chinese Journal of Veterinary Drug, 2011, 45(7):41-44. (in Chinese)
[8]	KUMAR N, SINGH N P. Effect of dietary selenium on immuno-biochemical plasticity and resistance against Aeromonas veronii biovar sobria in fish reared under multiple stressors[J]. Fish Shellfish Immunol, 2019, 84:38-47.
[9]	胡萌. 江苏地区气单胞菌分离鉴定及强毒株生物学特性分析[D]. 南京:南京农业大学, 2012.HU M. Isolation and identification of Aeromonas from Jiangsu and characterizations of virulent strains[D]. Nanjing:Nanjing Agricultural University, 2012. (in Chinese)
[10]	ELABD H, WANG H P, SHAHEEN A, et al. Astragalus membranaceus nanoparticles markedly improve immune and anti-oxidative responses; and protection against Aeromonas veronii in Nile tilapia Oreochromis niloticus[J]. Fish Shellfish Immunol, 2020, 97:248-256.
[11]	HOAI T D, TRANG T T, VAN TUYEN N, et al. Aeromonas veronii caused disease and mortality in channel catfish in Vietnam[J]. Aquaculture, 2019, 513:734425.
[12]	龚倩, 高淑琴, 单晓枫, 等. 框镜鲤致病性维氏气单胞菌的分离鉴定[J]. 中国预防兽医学报, 2010, 32(12):981-983.GONG Q, GAO S Q, SHAN X F, et al. Isolation and identification of pathogenic Aeromonas veronii from Cyprinus carpio[J]. Chinese Journal of Preventive Veterinary Medicine, 2010, 32(12):981-983. (in Chinese)
[13]	李槿年, 马彤彤, 房慧, 等. 草鱼源致病性维氏气单胞菌的鉴定及其黏附特性分析[J]. 安徽农业大学学报, 2019, 46(4):603-609.LI J N, MA T T, FANG H, et al. Identification of pathogenic Aeromonas veronii isolated from grass carps (Ctenopharyngodon idella) and analysis of its adhesion characteristics[J]. Journal of Anhui Agricultural University, 2019, 46(4):603-609. (in Chinese)
[14]	胡天野. 健康乌鳢体内两种气单胞菌的分离、鉴定及其毒力基因检测[D]. 长春:吉林农业大学, 2013.HU T Y. Isolation, identification and virulence genes detection of two kinds of Aeromonas isolated from healthy Channa argus[D]. Changchun:Jilin Agricultural University, 2013. (in Chinese)
[15]	EL LATIF A M A, ELABD H, AMIN A, et al. High mortalities caused by Aeromonas veronii:identification, pathogenicity, and histopathologicalstudies in Oreochromis niloticus[J]. Aquaculture International, 2019, 27(6):1725-1737.
[16]	朱成科, 向桢, 叶华, 等. 黄颡鱼致病性维氏气单胞菌的分离鉴定[J]. 西南大学学报:自然科学版, 2013, 35(5):37-42.ZHU C K, XIANG Z, YE H, et al. Isolation and identification of pathogenic Aeromonas veronii from Yellow Catfish Pelteobagrus fulvidraco[J]. Journal of Southwest University:Natural Science Edition, 2013, 35(5):37-42. (in Chinese)
[17]	DE S. JAGODA S S S, HONEIN K, ARULKANTHAN A, et al. Genome sequencing and annotation of Aeromonas veronii strain Ae52, a multidrug- resistant isolate from septicaemic gold fish (Carassius auratus) in Sri Lanka[J]. Genom Data, 2017, 11:46-48.
[18]	陆梦莹, 胡秀彩, 吕爱军, 等. 鲶源维氏气单胞菌的分离鉴定及药敏特性[J]. 大连海洋大学学报, 2017, 32(5):563-567.LU M Y, HU X C, LÜ A J, et al. Isolation, identification and susceptibility of Aeromonas veronii from diseased northern sheatfish Silurus soldatovi[J]. Journal of Dalian Ocean University, 2017, 32(5):563-567. (in Chinese)
[19]	黎炯, 叶星, 卢迈新, 等. 罗非鱼维氏气单胞菌的分离鉴定和药敏试验[J]. 水生态学杂志, 2011, 32(3):132-136.LI J, YE X, LU M X, et al. Isolation and identification of Aeromonas veronii from infected tilapia fry and analysis of its drug sensitivity[J]. Journal of Hydroecology, 2011, 32(3):132-136. (in Chinese)
[20]	任燕, 高彩霞, 曾伟伟, 等. 两株草鱼源维氏气单胞菌菌株的主要表型特征及致病力比较[J]. 中国预防兽医学报, 2019, 41(2):145-150.REN Y, GAO C X, ZENG W W, et al. Pathogenicity and main phenotype characteristics of two Aeromonas veronii isolated from diseased grass carp[J]. Chinese Journal of Preventive Veterinary Medicine, 2019, 41(2):145-150. (in Chinese)
[21]	张飘, 胡安东, 杨霞, 等. 一株草鱼维氏气单胞菌的分离鉴定及毒力基因分析[J]. 中国畜牧兽医, 2020, 47(3):848-855.ZHANG P, HU A D, YANG X, et al. Isolation identification and virulence gene analysis of a grass carp Aeromonas veronii[J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(3):848-855. (in Chinese)
[22]	JU A Q, YANG S B, ZHANG H P, et al. Construction and immune efficacy of recombinant Lactobacillus casei strains expressing Malt from Aeromonas veronii[J]. Microb Pathog, 2020, 141:103918.
[23]	FOYSAL J, MOMTAZ F, ALI H, et al. Molecular characterization and interactome analysis of aerolysin (aer) gene from fish pathogen Aeromonas veronii:the pathogenicity inferred from sequence divergence and linked to histidine kinase (che A)[J]. J Fish Dis, 2019, 42(4):465-475.
[24]	朱利霞, 王洪彬, 赵希艳, 等. 水貂源肺炎克雷伯菌分离鉴定与药敏试验[J]. 动物医学进展, 2019, 40(9):122-127.ZHU L X, WANG H B, ZHAO X Y, et al. Isolation, identification and drug susceptibility test of Klebsiella pneumoniae in mink[J]. Progress in Veterinary Medicine, 2019, 40(9):122-127. (in Chinese)
[25]	SREEDHARAN K, PHILIP R, SINGH I S B. Characterization and virulence potential of phenotypically diverse Aeromonas veronii isolates recovered from moribund freshwater ornamental fishes of Kerala, India[J]. Antonie Van Leeuwenhoek, 2013, 103(1):53-67.
[26]	YANG B T, SONG H C, AN D J, et al. Functional analysis of preA in Aeromonas veronii TH0426 reveals a key role in the regulation of virulence and resistance to oxidative stress[J]. Int J Mol Sci, 2020, 21(1):98.
[27]	胡安东, 杨霞, 张飘, 等. 水生动物维氏气单胞菌病概述[J]. 贵州畜牧兽医, 2019, 43(2):39-42.HU A D, YANG X, ZHANG P, et al. Summary of aquatic animals with Aeromonas veronii[J]. Guizhou Journal of Animal Husbandry & Veterinary Medicine, 2019, 43(2):39-42. (in Chinese)
[28]	TAO L T, KANG Y H, ZHANG L, et al. Cadaverine reverse transporter (CadB protein) contributes to the virulence of Aeromonas veronii TH0426[J]. Int Microbiol, 2020, doi:10. 1007/s10123-020-00120-6.
[29]	康元环, 张冬星, 杨滨僮, 等. 维氏气单胞菌最新研究进展[J]. 中国人兽共患病学报, 2018, 34(5):452-459, 465.KANG Y H, ZHANG D X, YANG B T, et al. Latest research progress on Aeromonas veronii[J]. Chinese Journal of Zoonoses, 2018, 34(5):452-459, 465. (in Chinese)
[30]	CHEN C, YANG B T, RAZA S H A, et al. Role of Myeloperoxidase of northern snakehead (Channa argus) in Aeromonas veronii infection[J]. Microb Pathog, 2019, 135:103622.

Isolation, Identification and Drug Sensitivity Test of a Strain of Aeromonas veronii from Chinemys reevesii

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 30

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	CHEN Yuetong, LIU Xiaohan, WANG Zhiyang, ZHAO Yuxin, ZHOU Tiezhong, HU Zengjin, ZHU Yue, WANG Shaohui, TIAN Mingxing, DING Siyu, QI Jingjing, YU Shengqing. Isolation, Identification, Pathogenicity and Drug Susceptibility of Mycoplasma gallisepticum from Dead Chicken Embryos in Large-scale Chicken Farms in Guangdong Province [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 290-299.
[2]	LIU Xinhuan, YUN Jialei, MAO Li, LI Jizong, HAO Fei, HE Miaofeng, YANG Leilei, ZHANG Wenwen, CHENG Zilong, SUN Min, LIU Maojun, WANG Shaohui, BAI Juan, LI Wenliang. Isolation, Identification, Virulence Genes and Drug Resistance Analysis of Escherichia coli Isolated from Diarrheal Goat and Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3445-3454.
[3]	WANG Jiali, ZHOU Ning, CHEN Xi, YUE Hua, TANG Cheng. Isolation, Identification and Pathogenicity of Canine Adenovirus Type 2 Strain with Natural Deletion in E3 Gene [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2982-2990.
[4]	ZHAO Feifei, LI Jie, HAN Ning, XIE Shiting, ZENG Zhenling. Antibacterial Drug Resistance Analysis of Klebsiella pneumoniae Isolated from Slaughterhouse [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 3044-3053.
[5]	YUAN Sheng, LI Anqi, Lü Wenke, YANG Lulu, ZHOU Feng, HUANG Liangzong, BAI Aiquan, WEN Feng, HUANG Shujian, GUO Jinyue. Analysis of Variation in Major Virulence-related Genes of a Strain of Pseudorabies Virus and Its Pathogenicity to Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2195-2199.
[6]	LIU Jiankui, XU Ye, LIU Chen, YU Hui, YANG Yuan, HE Le, LI Jiarui, DAN Huijuan, DAI Ailing, YANG Xiaoyan, WEI Chunhua. Genomic Characteristics of Porcine Reproductive and Respiratory Syndrome Virus in Fujian Province from 2017 to 2021 based on Whole Genome [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1579-1589.
[7]	JIANG Zenghai, TENG Lin, HE Anwen, LIU Yanyan, YUE Min, HE Qigai. Genomic Analysis of Salmonella Typhimurium Isolates and Salmonella Serotype 4, [5], 12: i:- Isolates from Pig-borne Food Chain [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1199-1209.
[8]	WANG Jianing, ZHANG Ziqiang, KONG Dejing, FENG Caicai, ZHANG Feike, LIU Yumei. Isolation and Identification of Klebsiella pneumoniae in Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5198-5206.
[9]	ZHANG Ao, TAN Bin, LIU Kexin, LIU Jiali, ZHANG Shuqin. Genomic Characterization Analysis of a H1N1 Subtype Swine Influenza Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4866-4871.
[10]	YANG Menglin, ZHENG Shiqi, PENG Kai, WANG Wei, HUANG Yanhua, PENG Jie. Isolation and Identification of Pigeon-derived Salmonella Typhimurium and Pathogenic Analysis [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4880-4888.
[11]	LI Chang, ZHOU Ping, WEN Ping, YANG Keli, LIU Wei, GUO Rui, LIU Zewen, YUAN Fangyan, GAO Ting, SONG Haofei, TIAN Yongxiang, ZHOU Danna. Investigation of the Genome-wide Sequence and Immunogenicity of Capsid Protein in Mice for a Porcine Circovirus 2d Genotype Strain [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4289-4299.
[12]	ZHANG Kaichuan, WANG Jinyu, LI Shoujun, JIA Kun. Isolation, Identification and Biological Characteristics of Klebsiella pneumoniae from Sheep in Guangdong Province [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 328-337.
[13]	WANG Meng, ZHANG Senhao, ZHANG Liu, LI Jiaxuan, WANG Xiaona, CUI Wen, JIANG Yanping, ZHOU Han, WANG Li, QIAO Xinyuan, LI Yijing, TANG Lijie. Isolation,Identification and Genetic Evolution Analysis of a Subgroup E Avian Leukemia Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3107-3120.
[14]	XU Lifeng, JIA Chenyu, CHEN Fuzai, FANG Mengyuan, LIU Xiaodan, CHEN Jilong, LI Xunliang. Isolation, Identification and Whole Genome Analysis of a Mycoplasma synoviae Strain from Chicken [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2663-2676.
[15]	WANG Xi, LI Ke, LI Tingcui, YAN Hongya, ZHAO Rong, CHANG Zhishun, LIAO Ming, SUN Minhua, XIN Aiguo. MLST Typing and Drug Resistance Analysis of 75 Salmonella Strains Isolated from Laying Hens [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(5): 1626-1631.