瑶山亚种树鼩源嗜水性气单胞菌的分离鉴定及生物学特性研究

doi:10.16431/j.cnki.1671-7236.2020.05.031

Abstract

Abstract: In order to confirm the cause of death of Tupaia belangeri yaoshanensis in the artificial domestication and breeding experiment center,the pathogenicity and drug resistance of the isolated strain were investigated.Pathogenic bacteria was isolated and obtained by plate drawing,and further morphological observation,culture characteristics analysis,biochemical test and 16S rRNA sequence analysis were carried out on the isolated bacteria.A pathogenic Aeromonas hydrophila was purified and obtained.The lethal dose 50%(LD₅₀) of the isolated strain was determined by pathogenicity test on Kunming mice.At the same time,the sensitivity of Aeromonas hydrophila isolates to common drugs was analyzed using the paper sensitive method,and the drug resistance genes of the isolates were detected by PCR.The results showed that a strain of Aeromonas hydrophila was isolated from the diseased tree shrews and the microscopic examination showed Gram-negative bacteria in a short rod shape.The 16S rRNA sequence of this strain showed that a strain of Aeromonas hydrophila was isolated from the diseased tree shrew,which was 100% homologous to the Aeromonas aeruginosa collected by GenBank SWCY-3.27 isolated from freshwater lake,and had the closest genetic evolution.The LD₅₀ of the isolates to adult Kunming mice was 1×10⁷ CFU,which was more virulent than the model strain ATCC 7966 and similar to the prevalent strains NJ-35 and J-1,etc.in China.The pathogenicity test identified the isolates as highly virulent.The biochemical identification of esculin,arabinose and oxidase was positive,and various biochemical identifications were consistent with the biochemical characteristics of Aeromonas hydrophila.The drug sensitivity test showed that the strain had high sensitivity to 9 drugs such as minocycline,streptomycin and tobramycin,completely resistant to 10 drugs such as tetracycline,erythromycin and penicillin.PCR amplification was used to detect 12 drug resistance genes of 6 major categories in the isolated bacteria,and it was found that there were two antibiotic resistance genes of aminoglycoside (Aph-(3)-lia) and β-lactams (mecA) in the isolated bacteria,which were basically consistent with the drug sensitivity test results.This study indicated that the pathogen causing the death of tree shrew was Aeromonas hydrophila,which could be treated with high-sensitivity drugs,and provided reference for the prevention and treatment of tree shrew diseases caused by Aeromonas hydrophila infection.

Key words: Tupaia belangeri; Aeromonas hydrophila; 16S rRNA; pathogenicity; drug resistance gene

CLC Number:

S852.61⁺7

TANG Haibo, YUN Chenxia, LIANG Liang, YUAN Xiaofang, HUANG Zhengtuan, CAO Yinyin, LENG Jing. Isolation and Identification and Biological Characterization of Aeromonas hydrophila from Tupaia belangeri yaoshanensis[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(5): 1560-1570.

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Isolation and Identification and Biological Characterization of Aeromonas hydrophila from Tupaia belangeri yaoshanensis

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