鸭源绿色气球菌的分离鉴定及16S rRNA基因序列测定与遗传进化分析

doi:10.16431/j.cnki.1671-7236.2022.02.004

Abstract

Abstract: 【Objective】 The experiment was aimed to study the pathogenic bacteria of acute death of ducklings in a duck farm in Henan province and its phylogenetic status.【Method】 The liver and spleen of dead ducks in the diseased duck farm were collected.The bacterial morphology, Gram-staining, biochemical characteristics, sequence analysis of 16S rRNA gene, drug sensitivity test and pathogenicity test were carried out.【Results】 The isolated bacteria grew on blood agar medium with smooth, convex, milky white and α-hemolytic Gram-positive cocci.Biochemical test results showed that the isolated bacteria were positive for maltose, sucrose, raffinose, nitrate reduction reaction, MP-VP test, urea, lysine decarboxylase, ornithine decarboxylase and aescin, and negative for xylose, lactose, glucose, sorbitol, hydrogen sulfide, citrate, peptone, mannitol, phenylalanine and methyl red.The results of 16S rRNA gene sequencing and BLAST comparison results showed that the 16S rRNA gene sequence of the isolated strain was 94.8%-99.9% similar to that of Aerococcus viridans published on NCBI, and the similarity with strains Mnlv1, Mnlv2 and W66 was the highest, up to 99.9%.The similarity with strain GXBl-1 was the lowest, which was 94.8%.Phylogenetic tree analysis showed that the isolated bacteria belonged to the same genus and group on the same branch as strains of Aerococcus viridans FL09, 15MS and Mnlv2 etc., and had the closest genetic relationship with FL09.The results of drug sensitivity showed that the isolated strain was sensitive to fosfomycin, ampicillin and amoxicillin, moderately sensitive to penicillin, neomycin and amikacin, it was resistant to rifampicin, neomycin, bacitracin, erythromycin and enrofloxacin.The results of pathogenicity test showed that the strain was lethal and increased with the increase of inoculation concentration.【Conclusion】 This study determined that the pathogen causing the acute death of ducklings in a duck farm in Henan province was Aerococcus viridans, which provided a reference basis for clinical diagnosis and treatment of the diseases caused by this bacteria.

Key words: duck; Aerococcus viridans; isolation and identification; 16S rRNA; genetic evolution

CLC Number:

S852.61⁺1

QIAO Hongxing, DING Yong, ZHANG Liheng, SONG Yuzhen, DONG Qing, BIAN Chuanzhou, ZHAO Junqiang. Isolation, Identification, 16S rRNA Gene Sequencing and Genetic Evolution Analysis of Aeromonas aeruginosa from Duck[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 435-442.

References

[1] WILLIAMS R E, HIRCH A, COWAN S T. Aerococcus, a new bacterial genus[J]. Journal of General Microbiology, 1953, 8(3): 475-480.
[2] 李金钟, 刘利平. 气球菌分类及鉴定进展[J]. 国外医学. 临床生物化学与检验学分册, 2005, 1: 35-37.
LI J Z, LIU L P. Progress in classification and identification of Aerococcus[J]. Foreign Medicine. Clinical Biochemistry and Laboratory Science Volume, 2005, 1: 35-37. (in Chinese)
[3] 袭恒豫. 绿色气球菌噬菌体AVP与其裂解酶的生物学特性及对小鼠乳腺炎模型的实验性治疗[D]. 长春: 吉林大学, 2020.
XI H Y. The biological characteristics of Aerococcus viridans phage AVP and its endolysin and their experimental treatment of mouse mastitis model[D]. Changchun: Jilin University, 2020. (in Chinese)
[4] YADAV K, SHARMA M, AGARWAL S, et al. Aortic pseudoaneurysm & endocarditis caused by Aerococcus viridans: A case report and literature review[J]. Cardiovascular Revascularization Medicine, 2018, 19(2): 201-203.
[5] 刘洁, 刘梦龙, 曲久燕, 等. 猪源绿色气球菌的分离与鉴定[J]. 中国猪业, 2019, 14(6): 26-30.
LIU J, LIU M L, QU J Y, et al. Isolation and identification of Aerococcus viridans from swine[J]. China Swine Industry, 2019, 14(6): 26-30. (in Chinese)
[6] AUGUSTINE T, THIRUNAVUKKARASU B, BHAT B V. Aerococcus viridans endocarditis. Case report[J]. Indian Pediatrics, 1994, 31(5): 599-601.
[7] POPESC G A, BENEA E, MITACHE E, et al. An unusual bacterium, Aerococcus viridans, and four cases of infective endocarditis[J]. Journal of Heart Valve Disease, 2005, 14(3): 317-390.
[8] CATTOIR V, KOBAL A, LEGRAND P. Aerococcus urinae and Aerococcus sanguinicola, two frequently misidentified uropathogens[J]. Scandinavian Journal of Infectious Diseases, 2010, 42(10): 775-780.
[9] 张乐宜, 李艳, 蔡汝健, 等. 猪源绿色气球菌的分离鉴定及16S rRNA基因序列测定与遗传进化分析[J]. 中国农学通报, 2014, 30(29): 15-21.
ZHANG L Y, LI Y, CAI R J, et al. Isolation and identification of Aerococcus viridans from swine and phylogenetic analysis of 16S rRNA gene sequence[J]. Chinese Agricultural Science Bulletin, 2014, 30(29): 15-21. (in Chinese)
[10] 李楠, 刘晔, 张守峰, 等. 一株山羊源浅绿色气球菌的分离[J]. 黑龙江畜牧兽医, 2015, 18: 109-111.
LI N, LIU Y, ZHANG S F, et al. Isolation of a strain of goat-derived Aerococcus viridans[J]. Heilongjiang Animal Science and Veterinary Medicine, 2015, 18: 109-111. (in Chinese)
[11] LIU G, LIU Y, ALI T, et al. Molecular and phenotypic characterization of Aerococcus viridans associated with subclinical bovine mastitis[J]. The Public Library of Science, 2015, 10(4): e0125001.
[12] 朱明华, 朱瑞良, 马荣德. 鸡奇异变形杆菌的分离鉴定和16S rRNA基因序列测定与系统进化分析[J]. 中国兽医学报, 2011, 31(6): 804-808.
ZHU M H, ZHU R L, MA R D, et al. Isolation, identification of chicken Proteus mirabilis and phylogenetic analysis of 16S rRNA gene sequence[J]. Chinese Journal of Veterinary Science, 2011, 31(6): 804-808. (in Chinese)
[13] 阎朝华, 文明, 周碧君, 等. 竹鼠源绿色气球菌的分离鉴定及耐药性分析[J]. 中国畜牧兽医, 2020, 47(8): 2643-2651.
YAN C H, WEN M, ZHOU B J, et al. Isolation, identification and drug resistance analysis of Aerococcus viridans from Rhizomys sinensis[J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(8): 2643-2651. (in Chinese)
[14] 可小丽, 卢迈新, 黎炯, 等. 罗非鱼绿色气球菌的鉴定及致病性研究[J]. 水生生物学报, 2011, 35(5): 796-802.
KE X L, LU M X, LI J, et al. Identification and pathogenicity of tilapia Aerococcus viridans bacteria[J]. Acta Hydrobiologica Sinica, 2011, 35(5): 796-802. (in Chinese)
[15] GREENWOOD S J, KEITH I R, DESPRES B M, et al. Genetic characterization of the lobster pathogen Aerococcus viridans var. homari by 16S rRNA gene sequence and RAPD[J]. Diseases of Aquatic Organisms, 2005, 63(2-3): 237-246.
[16] STEWART J E, CORNICK J W, ZWICKER B M, et al. Studies on the virulence of Aerococcus viridans (var. ) homari, the causative agent of gaffkemia, a fatal disease of homarid lobsters[J]. Diseases of Aquatic Organisms, 2004, 60(2): 149-155.
[17] GUTIÉRREZ-FERNÁNDEZ J, GÁMIZ-GÁMIZ A, NAVARRO-MARÍ. J M, et al. Genitourinary tract infection in children due to Aerococcus other than Aerococcus viridans. Literature review and 3 case reports[J]. Enfermedades Infecciosasy Microbiologia Clinica, 2021, 39(3): 156-158.
[18] UH Y, SON J S, JANG I H, et al. Penicillin-resistant Aerococcus viridans bacteremia associated with granulocytopenia[J]. Journal Korean Medical Science, 2002, 17(1): 113-115.
[19] 付琦媛, 严世杰, 宋涛, 等. 猪源绿色气球菌的分离鉴定及耐药性分析[J]. 中国兽医学报, 2020, 40(5): 943-946.
FU Q Y, YAN S J, SONG T, et al. Isolation, identification and antibiotic resistance of Aerococcus viridans from swine[J]. Chinese Journal of Veterinary Science, 2020, 40(5): 943-946. (in Chinese)
[20] 张乃嘉, 李建波, 陈宏达, 等. 猪源绿色气球菌的分离与鉴定[J]. 黑龙江畜牧兽医, 2018, 9: 130-134.
ZHANG N J, LI J B, CHEN H D, et al. Isolation and identification of Aerococcus viridans from swine[J]. Heilongjiang Animal Science and Veterinary Medicine, 2018, 9: 130-134. (in Chinese)
[21] 董文龙, 王巍, 勾长龙, 等. 6株猪源绿色气球菌的分离鉴定及耐药性分析[J]. 中国兽医学报, 2017, 37(12): 2256-2259.
DONG W L, WANG W, GOU C L, et al. Isolation, identification and drug resistance analysis of 6 Aerococcus viridans from swine[J]. Chinese Journal of Veterinary Science, 2017, 37(12): 2256-2259. (in Chinese)
[22] CHEN L Y, YU W C, HUANG S H, et al. Successful treatment of Aerococcus viridans endocarditis in a patient allergic to penicillin[J]. Journal of Microbiology Immunology and Infection, 2012, 45(2): 158-160.
[23] 阚威, 马艳萍, 张总超, 等. 2株猪源性绿色气球菌的分离鉴定及药敏特性研究[J]. 畜牧兽医杂志, 2020, 39(4): 6-11.
KAN W, MA Y P, ZHANG Z C, et al. Isolation, identification and drug susceptibility test of two strains of Aerococcus viridans from swine[J]. Journal of Animal Science and Veterinary Medicine, 2020, 39(4): 6-11. (in Chinese)

Isolation, Identification, 16S rRNA Gene Sequencing and Genetic Evolution Analysis of Aeromonas aeruginosa from Duck

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