副猪嗜血杆菌qseB、qseC双基因缺失株的构建及生物学特性

doi:10.11843/j.issn.0366-6964.2022.02.019

Abstract

Abstract: The QseBC two-component system (TCS) is related to quorum sensing of bacteria. As a global regulator of virulence in Enterobacteriaceae and Pasteurellaceae, its role still unclear in Glaesserella (Haemophilus) parasuis. In this study, the double gene deletion mutant ΔqseBC of SC1401 was constructed by natural transformation method. The serum resistance ability, biofilm formation, antibiotic resistance and stress tolerance of ΔqseBC mutant strain and wild strain SC1401 were measured. Results were as follows:the biofilm-forming ability of ΔqseBC was significantly weaker than that of the wild strain SC1401. The survival rate of the ΔqseBC mutant in porcine serum was 55.12%, which was lower than that of the wild strain SC1401 (83.76%). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of ΔqseBC mutant strain to enrofloxacin, gentamicin, ceftiofur and penicillin were significantly decreased. The ΔqseBC mutant strain showed high sensitivity to high temperature (39 and 42℃), hydrogen peroxide (1, 2, 4, 8 and 16 mmol·L^-1) and NaCl (50, 100 and 150 mmol·L^-1). The results showed that the TCS QseBC plays an important role in the serum resistance, antibiotic resistance and stress tolerance of Glaesserella parasuis. The above findings revealed that QseBC might be closely related to the pathogenicity of Glaesserella parasuis, but further studies are needed to determine the specific mechanism.

Key words: QseBC, serum resistance, biofilm, antibiotic resistance, stress tolerance

CLC Number:

S852.61

HE Lüqin, YAN Xuefeng, WEN Xintian, CAO Sanjie, HUANG Xiaobo, WU Rui, ZHAO Qin, WEN Yiping. Construction and Biological Characteristic of Glaesserella parasuis Strain with qseB, qseC Double Gene Deletion[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 529-537.

References 39

[1]	LI J, PENG H, XU L G, et al. Draft genome sequence of Haemophilus parasuis gx033, a serotype 4 strain isolated from the swine lower respiratory tract[J]. Genome Announc, 2013, 1(3):e00224-13.
[2]	OLIVEIRA S, PIJOAN C. Haemophilus parasuis:new trends on diagnosis, epidemiology and control[J]. Vet Microbiol, 2004, 99(1):1-12.
[3]	CAI X W, CHEN H C, BLACKALL P J, et al. Serological characterization of Haemophilus parasuis isolates from China[J]. Vet Microbiol, 2005, 111(3-4):231-236.
[4]	ZHANG L H, LI Y, DAI K, et al. Establishment of a successive markerless mutation system in Haemophilus parasuis through natural transformation[J]. PLoS One, 2015, 10(5):e0127393.
[5]	ESPÍNDOLA J P, BALBINOTT N, GRESSLER L T, et al. Molecular serotyping of clinical strains of Haemophilus (Glaesserella) parasuis brings new insights regarding Glässer's disease outbreaks in Brazil[J]. PeerJ, 2019, 7:e6817.
[6]	SHEN Y J, ZHOU N N, AN J H, et al. Haemophilus parasuis infection in 3D4/21 cells induces autophagy through the AMPK pathway[J]. Cell Microbiol, 2019, 21(8):e13031.
[7]	HUANG J C, WANG X R, CAO Q, et al. ClpP participates in stress tolerance and negatively regulates biofilm formation in Haemophilus parasuis[J]. Vet Microbiol, 2016, 182:141-149.
[8]	STOCK A M, ROBINSON V L, GOUDREAU P N. Two-component signal transduction[J]. Annu Rev Biochem, 2000, 69:183-215.
[9]	APPLEBY J L, PARKINSON J S, BOURRET R B. Signal transduction via the multi-step phosphorelay:not necessarily a road less traveled[J]. Cell, 1996, 86(6):845-848.
[10]	HE L Q, DAI K, WEN X T, et al. QseC mediates osmotic stress resistance and biofilm formation in Haemophilus parasuis[J]. Front Microbiol, 2018, 9:212.
[11]	KOSTAKIOTI M, HADJIFRANGISKOU M, PINKNER J S, et al. QseC-mediated dephosphorylation of QseB is required for expression of genes associated with virulence in uropathogenic Escherichia coli[J]. Mol Microbiol, 2009, 73(6):1020-1031.
[12]	LIU J L, HU L L, XU Z F, et al. Actinobacillus pleuropneumoniae two-component system QseB/QseC regulates the transcription of PilM, an important determinant of bacterial adherence and virulence[J]. Vet Microbiol, 2015, 177(1-2):184-192.
[13]	LUO Z, WANG M, DU H, et al. The putative sensor kinase QseC of Salmonella enterica serovar Typhi can promote invasion in the presence of glucose[J]. Food Res Int, 2012, 45(2):1004-1010.
[14]	VNAL C M, SINGH B, FLEURY C, et al. QseC controls biofilm formation of non-typeable Haemophilus influenzae in addition to an AI-2-dependent mechanism[J]. Int J Med Microbiol, 2012, 302(6):261-269.
[15]	XU J, FU S L, LIU M L, et al. The two-component system NisK/NisR contributes to the virulence of Streptococcus suis serotype 2[J]. Microbiol Res, 2014, 169(7-8):541-546.
[16]	WEIGEL W A, DEMUTH D R. QseBC, a two-component bacterial adrenergic receptor and global regulator of virulence in Enterobacteriaceae and Pasteurellaceae[J]. Mol Oral Microbiol, 2016, 31(5):379-397.
[17]	BEARSON B L, BEARSON S M D. The role of the QseC quorum-sensing sensor kinase in colonization and norepinephrine-enhanced motility of Salmonella enterica serovar Typhimurium[J]. Microb Pathog, 2008, 44(4):271-278.
[18]	SPERANDIO V, TORRES A G, KAPER J B. Quorum sensing Escherichia coli regulators B and C (QseBC):a novel two-component regulatory system involved in the regulation of flagella and motility by quorum sensing in E. coli[J]. Mol Microbiol, 2002, 43(3):809-821.
[19]	CLARKE M B, HUGHES D T, ZHU C R, et al. The QseC sensor kinase:a bacterial adrenergic receptor[J]. Proc Natl Acad Sci U S A, 2006, 103(27):10420-10425.
[20]	HUGHES D T, CLARKE M B, YAMAMOTO K, et al. The QseC adrenergic signaling cascade in Enterohemorrhagic E. coli (EHEC)[J]. PLoS Pathog, 2009, 5(8):e1000553.
[21]	JI Y, LI W L, ZHANG Y, et al. QseB mediates biofilm formation and invasion in Salmonella enterica serovar Typhi[J]. Microb Pathog, 2017, 104:6-11.
[22]	何绿琴. 副猪嗜血杆菌双组份系统QseB/QseC调控功能研究[D]. 成都:四川农业大学, 2018.HE L Q. Study on the regulatory function of two-component system QseB/QseC of Haemophilus parasuis[D]. Chengdu:Sichuan Agricultural University, 2018. (in Chinese)
[23]	ZOU Y, FENG S X, XU C G, et al. The role of galU and galE of Haemophilus parasuis SC096 in serum resistance and biofilm formation[J]. Vet Microbiol, 2013, 162(1):278-284.
[24]	BLANCHETTE-CAIN K, HINOJOSA C A, AKULA SURESH BABU R, et al. Streptococcus pneumoniae biofilm formation is strain dependent, multifactorial, and associated with reduced invasiveness and immunoreactivity during colonization[J]. mBio, 2013, 15;4(5):e00745-13.doi:10.1128/mBio.00745-13.
[25]	MORIYAMA S, HOTOMI M, SHIMADA J, et al. Formation of biofilm by Haemophilus influenzae isolated from pediatric intractable otitis media[J]. Auris Nasus Larynx, 2009, 36(5):525-531.
[26]	杨开杰, 冯赛祥, 周琪, 等. 副猪嗜血杆菌qseC基因缺失突变对生物被膜形成的影响[J]. 畜牧兽医学报, 2017, 48(8):1499-1504.YANG K J, FENG S X, ZHOU Q, et al. Effect of qseC gene mutation of Haemophilus parasuis on biofilm formation[J]. Acta Veterinaria et Zootechnica Sinica, 2017, 48(8):1499-1504. (in Chinese)
[27]	LI Y, CAO S J, ZHANG L H, et al. A TolC-like protein of Actinobacillus pleuropneumoniae is involved in antibiotic resistance and biofilm formation[J]. Front Microbiol, 2016, 7:1618.
[28]	ZHANG L, MAH T F. Involvement of a novel efflux system in biofilm-specific resistance to antibiotics[J]. J Bacteriol, 2008, 190(13):4447-4452.
[29]	HE L Q, WEN X T, YAN X F, et al. Effect of cheY deletion on growth and colonization in a Haemophilus parasuis serovar 13 clinical strain EP3[J]. Gene, 2016, 577(1):96-100.
[30]	DONLAN R M, COSTERTON J W. Biofilms:survival mechanisms of clinically relevant microorganisms[J]. Clin Microbiol Rev, 2002, 15(2):167-193.
[31]	STEWART P S, COSTERTON J W. Antibiotic resistance of bacteria in biofilms[J]. Lancet, 2001, 358(9276):135-138.
[32]	赵良友, 高雪丽, 刘超男, 等. 副猪嗜血杆菌vacJ基因缺失菌株构建及其生物学特性分析[J]. 华北农学报, 2017, 32(5):19-24.ZHAO L Y, GAO X L, LIU C N, et al. Construction and biological characterization of vacJ gene deletion strain of Haemophilus parasuis[J]. Acta Agriculturae Boreali-Sinica, 2017, 32(5):19-24. (in Chinese)
[33]	CAO Q, FENG F F, WANG H, et al. Haemophilus parasuis CpxRA two-component system confers bacterial tolerance to environmental stresses and macrolide resistance[J]. Microbiol Res, 2018, 206:177-185.
[34]	CERDÀ-CUÉLLAR M, ARAGON V. Serum-resistance in Haemophilus parasuis is associated with systemic disease in swine[J]. Vet J, 2008, 175(3):384-389.
[35]	DAI K, HE L Q, CHANG Y F, et al. Basic characterization of natural transformation in a highly transformable Haemophilus parasuis strain SC1401[J]. Front Cell Infect Microbiol, 2018, 8:32.
[36]	谨瑾, 张禄滑, 文心田, 等. 副猪嗜血杆菌potD基因缺失株的构建及其部分生物学特性[J]. 畜牧兽医学报, 2016, 47(11):2274-2279.JIN J, ZHANG L H, WEN X T, et al. Construction and characterization of a Haemophilus parasuis potD mutant strain[J]. Acta Veterinaria et Zootechnica Sinica, 2016, 47(11):2274-2279. (in Chinese)
[37]	DING L Q, WEN X T, HE L Q, et al. The arcA gene contributes to the serum resistance and virulence of Haemophilus parasuis serovar 13 clinical strain EP3[J]. Vet Microbiol, 2016, 196:67-71.
[38]	DAI K, YANG Z, CHANG Y F, et al. Construction of targeted and integrative promoter-reporter plasmids pDK-K and pDK-G to measure gene expression activity in Haemophilus parasuis[J]. Microb Pathog, 2019, 134:103565.
[39]	FENG S X, XU L N, XU C G, et al. Role of acrAB in antibiotic resistance of Haemophilus parasuis serovar 4[J]. Vet J, 2014, 202(1):191-194.

Construction and Biological Characteristic of Glaesserella parasuis Strain with qseB, qseC Double Gene Deletion

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