树鼩源碳青霉烯耐药性铜绿假单胞菌的分离鉴定及生物学特性研究

doi:10.16431/j.cnki.1671-7236.2024.04.039

Abstract

Abstract: 【Objective】 This experiment was aimed to determine the cause of death of Tupaia belangeri during the breeding process,analyze the biological characteristics,pathogenicity and drug resistance of the pathogen,and develop prevention and control methods.【Method】 Pathological dissection was performed on the dead Tupaia belangeri and the pathological changes of each major organ were recorded,including lung,liver,spleen and intestine.Single colonies were isolated from the main diseased organs using the streak plate method,and the morphology of the colonies was observed.At the same time,Gram staining was used to observe under the microscope,followed by biochemical and drug resistance tests,and pathogenicity test.Further analysis was conducted on the isolated strains using 16S rRNA gene sequencing and bacteria genome sequencing.【Result】 A carbapenem-resistant Pseudomonas aeruginosa strain was isolated from the spleen and intestine of the deceased Tupaia belangeri chinensis.The isolate presented a gray-white colony producing green water-soluble pigments in LB agar medium,and was Gram-negative under microscope.Biochemical tests further confirmed that it was a non-fermenting bacterium.Sequence evolution analysis showed that it had a high similarity of 99% with the soil source Pseudomonas aeruginosa F4 in GenBank.The drug sensitivity test results showed that the isolated strain had certain resistance to carbapenem antibiotics such as imipenem and meropenem,as well as quinolone antibiotics such as levofloxacin.The pathogenicity test result showed that the median lethal dose (LD₅₀) of the isolated strain to adult Kunming mice was 3.382×10⁸ CFU.The organs and tissues of dead mice were isolated for pathological observation,and inflammatory damage and necrosis of the lungs,liver,spleen,intestine,etc.were observed.Bacterial isolation was conducted again on various organs,and based on 16S rRNA sequence analysis,Pseudomonas aeruginosa could be found in the spleen,intestine and blood of dead mice,with high pathogenicity.The whole genome sequencing results confirmed that the isolate strain had genes related to carbapenem resistance and shared similar virulence factor genes with the highly pathogenic reference strain of Pseudomonas aeruginosa.【Conclusion】 This study isolated one carbapenem-resistant Pseudomonas aeruginosa from the deceased Tupaia belangeri,providing a new theoretical basis for the study of microbial pathogenicity and domestication of Tupaia belangeri.

Key words: Tupaia belangeri; Pseudomonas aeruginosa; drug resistance

CLC Number:

S852.61

MU Zhengrong, CHENG Qingqing, ZHANG Dongjie, ZHAI Shanshan, LI Zhuxin, TAO Junyu, LENG Jing. Isolation,Identification and Biological Characteristics of Carbapenem-resistant Pseudomonas aeruginosa from Tupaia belangeri[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1717-1728.

References

[1] CHUNG J,EISHA S,PARK S,et al.How three self-secreted biofilm exopolysaccharides of Pseudomonas aeruginosa,Psl,Pel,and alginate,can each be exploited for antibiotic adjuvant effects in cystic fibrosis lung infection[J].International Journal of Molecular Sciences,2023,24(10):8709.
[2] TSAO L H,HSIN C Y,LIU H Y,et al.Risk factors for healthcare-associated infection caused by carbapenem-resistant Pseudomonas aeruginosa[J].Journal of Microbiology,Immunology and Infection,2018,51(3):359-366.
[3] TENOVER F C,NICOLAU D P,GILL C M.Carbapenemase-producing Pseudomonas aeruginosa-An emerging challenge[J].Emerging Microbes & Infections,2022,11(1):811-814.
[4] HU Y,QING Y,CHEN J,et al.Prevalence,risk factors,and molecular epidemiology of intestinal carbapenem-resistant Pseudomonas aeruginosa[J].Microbiology Spectrum,2021,9(3):e01344-21.
[5] TACCONELLI E,CARRARA E,SAVOLDI A,et al.Discovery,research,and development of new antibiotics:The WHO priority list of antibiotic-resistant bacteria and tuberculosis[J].The Lancet Infectious Diseases,2018,18(3):318-327.
[6] FAN Y,HUANG Z Y,CAO C C,et al.Genome of the Chinese tree shrew[J].Nature Communications,2013,4(1):1426.
[7] ZHANG J,XIAO H,BI Y,et al.Characteristics of the tree shrew humoral immune system[J].Molecular Immunology,2020,127:175-185.
[8] ZHANG L,WU X,LIAO S,et al.Tree shrew (Tupaia belangeri chinensis),a novel non-obese animal model of non-alcoholic fatty liver disease[J].Biology Open,2016,5(10):1545-1552.
[9] TSUKIYAMA-KOHARA K,KOHARA M.Tupaia belangeri as an experimental animal model for viral infection[J].Experimental Animals,2014,63(4):367-374.
[10] GU W,TONG P,LIU C,et al.The characteristics of gut microbiota and commensal Enterobacteriaceae isolates in tree shrew (Tupaia belangeri)[J].BMC Microbiology, 2019,19(1):203.
[11] 唐海波,运晨霞,梁亮,等.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[J].中国畜牧兽医,2020,47(5):1560-1570. TANG H B,YUN C X,LIANG L,et al.Isolation and identification and biological characterization of Aeromonas hydrophila from Tupaia belangeri yaoshanensis[J].China Animal Husbandry & Veterinary Medicine,2020,47(5):1560-1570.(in Chinese)
[12] SAITOU N,NEI M.The Neighbor-Joining method:A new method for reconstructing phylogenetic trees[J].Molecular Biology and Evolution,1987,4(4):406-425.
[13] ZHANG Y,CAO W,TONERI M,et al.Toxicology and efficacy of tumor-targeting Salmonella Typhimurium A1-R compared to VNP 20009 in a syngeneic mouse tumor model in immunocompetent mice[J].Oncotarget,2017,8(33):54616-54628.
[14] YIN D,GUO Y,LI M,et al.Performance of VITEK 2,E-test,Kirby-Bauer disk diffusion,and modified Kirby-Bauer disk diffusion compared to reference broth microdilution for testing tigecycline susceptibility of carbapenem-resistant K.pneumoniae and A.baumannii in a multicenter study in China[J].European Journal of Clinical Microbiology & Infectious Diseases,2021,40(6):1149-1154.
[15] EDGAR R C.UPARSE:Highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.
[16] ALCOCK B P,HUYNH W,CHALIL R,et al.CARD 2023:Expanded curation,support for machine learning,and resistome prediction at the comprehensive antibiotic resistance database[J].Nucleic Acids Research,2023,51(D1):D690-D699.
[17] LIU B,ZHENG D,ZHOU S,et al.VFDB 2022:A general classification scheme for bacterial virulence factors[J].Nucleic Acids Research,2022,50(D1):D912-D917.
[18] JURADO-MARTÍN I,SAINZ-MEJÍAS M,MCCLEAN S. Pseudomonas aeruginosa:An audacious pathogen with an adaptable arsenal of virulence factors[J].International Journal of Molecular Sciences,2021,22(6):3128.
[19] LO Y L,CHEN C L,SHEN L,et al.Characterization of the role of global regulator FLIA in the pathophysiology of Pseudomonas aeruginosa infection[J]. Research in Microbiology,2018,169(3):135-144.
[20] NANDI T,ONG C,SINGH A P,et al.A genomic survey of positive selection in Burkholderia pseudomallei provides insights into the evolution of accidental virulence[J].PLoS Pathogens,2010,6(4):e1000845.
[21] QIAN Z,HUI P,HAN L,et al.Serotypes and virulence genes of Pseudomonas aeruginosa isolated from mink and its pathogenicity in mink[J].Microbial Pathogenesis,2020,139:103904.
[22] ABD EL-GHANY W A.Pseudomonas aeruginosa infection of avian origin:Zoonosis and one health implications[J]. Veterinary World,2021,14(8):2155-2159.
[23] LI G,LAI R,DUAN G,et al.Isolation and identification of symbiotic bacteria from the skin,mouth,and rectum of wild and captive tree shrews[J].Zoological Research,2014,35(6):492-499.
[24] BACHTA K E R,ALLEN J P,CHEUNG B H,et al.Systemic infection facilitates transmission of Pseudomonas aeruginosa in mice[J].Nature Communications,2020,11(1):543.
[25] LORUSSO A B,CARRARA J A,BARROSO C D N,et al.Role of efflux pumps on antimicrobial resistance in Pseudomonas aeruginosa[J]. International Journal of Molecular Sciences,2022,23(24):15779.
[26] 刘兴雨,毕春霞,韩璐,等.铜绿假单胞菌CRISPR/Cas系统的基因结构及其与耐药基因的关系[J].中国病原生物学杂志,2022,17(12):1399-1403. LIU X Y,BI C X,HAN L,et al.Analysis of the gene structure of Pseudomonas aeruginosa CRISPR/Cas system and its relationship with antibiotic resistance related genes[J].Journal of Pathogen Biology,2022,17(12):1399-1403.(in Chinese)
[27] LI S A,LEE W H,ZHANG Y.Two bacterial infection models in tree shrew for evaluating the efficacy of antimicrobial agents[J].Zoological Research,2012,33(1):1-6.

Isolation,Identification and Biological Characteristics of Carbapenem-resistant Pseudomonas aeruginosa from Tupaia belangeri

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