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Published in: Journal of Orthopaedic Surgery and Research 1/2019

Open Access 01-12-2019 | Osteomyelitis | Research article

Staphylococcus aureus biofilm organization modulated by YycFG two-component regulatory pathway

Authors: Shizhou Wu, Fuguo Huang, Hui Zhang, Lei Lei

Published in: Journal of Orthopaedic Surgery and Research | Issue 1/2019

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Abstract

Background

Staphylococcus aureus (S. aureus) infection accounts for more than 50% of the osteomyelitis cases. Currently, methicillin-resistant S. aureus (MRSA) strains present an urgent medical problem. The YycFG two-component regulatory system (TCS) can allow bacteria to rapidly adapt to physical, chemical, and biological stresses. To define the role of YycFG in modulation virulence of S. aureus in osteomyelitis, we isolated clinical MRSA strains and compared these with ATCC29213 methicillin-sensitive S. aureus (MSSA).

Methods

In the present study, 13 MRSA strains from chronic osteomyelitis tissues were isolated. The in-depth sequencing of 16S rRNA amplicons of the samples was conducted. Bacterial growth was monitored, and biofilm biomass was determined by crystal violet microtiter assay. Furthermore, quantitative RT-PCR analysis was adopted to identify the expression of yycF/G/H and icaA/D in MRSA and MSSA strains. Analysis of variance with one-way ANOVA was used for statistical analysis.

Results

The in-depth sequencing of 16S rRNA amplicons of the clinical samples indicated a polymicrobial infection, with the phylum Firmicutes made up 13% of the microbial population. The MRSA strains showed an accelerated growth rate compared to the MSSA strains. Of note, MRSA biofilms showed an accumulation of an intercellular polysaccharides matrix and enhanced biomass upon microscopic examination. Furthermore, MRSA strains had a higher expression of the yycF/G/H and icaA/D genes and adhesion force.

Conclusions

These data suggested the roles of intercellular polysaccharide in S. aureus pathogenesis, indicating a possible association between YycFG pathways and MRSA strain virulence.
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Literature
1.
go back to reference Brady RA, Leid JG, Costerton JW, Shirtliff ME. Osteomyelitis: clinical overview and mechanisms of infection persistence. Clin Micro Newsletter. 2006;28(9):65–72.CrossRef Brady RA, Leid JG, Costerton JW, Shirtliff ME. Osteomyelitis: clinical overview and mechanisms of infection persistence. Clin Micro Newsletter. 2006;28(9):65–72.CrossRef
2.
go back to reference Beck-Broichsitter BE, Smeets R, Heiland M. Current concepts in pathogenesis of acute and chronic osteomyelitis. Curr Opin Infect Dis. 2015;28(3):240–5.CrossRef Beck-Broichsitter BE, Smeets R, Heiland M. Current concepts in pathogenesis of acute and chronic osteomyelitis. Curr Opin Infect Dis. 2015;28(3):240–5.CrossRef
4.
go back to reference Roberts ME, Stewart PS. Modeling antibiotic tolerance in biofilms by accounting for nutrient limitation. Antimicrob Agents Chemother. 2004;48(1):48–52.CrossRef Roberts ME, Stewart PS. Modeling antibiotic tolerance in biofilms by accounting for nutrient limitation. Antimicrob Agents Chemother. 2004;48(1):48–52.CrossRef
5.
go back to reference Mruk AL, Record KE. Antimicrobial options in the treatment of adult staphylococcal bone and joint infections in an era of drug shortages. Orthopedics. 2012;35(5):401–7.CrossRef Mruk AL, Record KE. Antimicrobial options in the treatment of adult staphylococcal bone and joint infections in an era of drug shortages. Orthopedics. 2012;35(5):401–7.CrossRef
6.
go back to reference Vardakas KZ, Kontopidis I, Gkegkes ID, Rafailidis PI, Falagas ME. Incidence, characteristics, and outcomes of patients with bone and joint infections due to community-associated methicillin-resistant Staphylococcus aureus: a systematic review. Eur J Clin Microbiol Infect Dis. 2013;32(6):711–21.CrossRef Vardakas KZ, Kontopidis I, Gkegkes ID, Rafailidis PI, Falagas ME. Incidence, characteristics, and outcomes of patients with bone and joint infections due to community-associated methicillin-resistant Staphylococcus aureus: a systematic review. Eur J Clin Microbiol Infect Dis. 2013;32(6):711–21.CrossRef
7.
go back to reference Ando E, Monden K, Mitsuhata R, Kariyama R, Kumon H. Biofilm formation among methicillin-resistant Staphylococcus aureus isolates from patients with urinary tract infection. Acta Med Okayama. 2004;58(4):207–14.PubMed Ando E, Monden K, Mitsuhata R, Kariyama R, Kumon H. Biofilm formation among methicillin-resistant Staphylococcus aureus isolates from patients with urinary tract infection. Acta Med Okayama. 2004;58(4):207–14.PubMed
8.
go back to reference Gómez-Mejia A, Gámez G, Hammerschmidt S. Streptococcus pneumoniae two-component regulatory systems: the interplay of the pneumococcus with its environment. Int J Med Microbiol. 2018;308(6):722–37.CrossRef Gómez-Mejia A, Gámez G, Hammerschmidt S. Streptococcus pneumoniae two-component regulatory systems: the interplay of the pneumococcus with its environment. Int J Med Microbiol. 2018;308(6):722–37.CrossRef
9.
go back to reference Dubrac S, Bisicchia P, Devine KM, Msadek T. A matter of life and death: cell wall homeostasis and the WalKR (YycGF) essential signal transduction pathway. Mol Microbiol. 2008;70(6):1307–22.CrossRef Dubrac S, Bisicchia P, Devine KM, Msadek T. A matter of life and death: cell wall homeostasis and the WalKR (YycGF) essential signal transduction pathway. Mol Microbiol. 2008;70(6):1307–22.CrossRef
10.
go back to reference Szurmant H, Mohan MA, Imus PM, Hoch JA. YycH and YycI interact to regulate the essential YycFG two-component system in Bacillus subtilis. J Bacteriol. 2007;189(8):3280–9.CrossRef Szurmant H, Mohan MA, Imus PM, Hoch JA. YycH and YycI interact to regulate the essential YycFG two-component system in Bacillus subtilis. J Bacteriol. 2007;189(8):3280–9.CrossRef
11.
go back to reference Archer NK, Mazaitis MJ, Costerton JW, Leid JG, Powers ME, Shirtliff ME. Staphylococcus aureus biofilms: properties, regulation, and roles in human disease. Virulence. 2011;2(5):445–59.CrossRef Archer NK, Mazaitis MJ, Costerton JW, Leid JG, Powers ME, Shirtliff ME. Staphylococcus aureus biofilms: properties, regulation, and roles in human disease. Virulence. 2011;2(5):445–59.CrossRef
12.
go back to reference Cramton SE, Ulrich M, Götz F, Döring G. Anaerobic conditions induce expression of polysaccharide intercellular adhesin in Staphylococcus aureus and Staphylococcus epidermidis. Infect Immun. 2001;69(6):4079–85.CrossRef Cramton SE, Ulrich M, Götz F, Döring G. Anaerobic conditions induce expression of polysaccharide intercellular adhesin in Staphylococcus aureus and Staphylococcus epidermidis. Infect Immun. 2001;69(6):4079–85.CrossRef
13.
go back to reference Pam B, Honarmand JS, Zare KS. Phenotypic and genotypic characterization of biofilm formation among Staphylococcus aureus isolates from clinical specimens, an atomic force microscopic (AFM) study. Microb Pathog. 2017;110:533–9.CrossRef Pam B, Honarmand JS, Zare KS. Phenotypic and genotypic characterization of biofilm formation among Staphylococcus aureus isolates from clinical specimens, an atomic force microscopic (AFM) study. Microb Pathog. 2017;110:533–9.CrossRef
14.
go back to reference Spangehl MJ, Masri BA, O’Connell JX, Duncan CP. Prospective analysis of preoperative and intraoperative investigations for the diagnosis of infection at the sites of two hundred and two revision total hip arthroplasties. J Bone Joint Surg Am. 1999;81(5):672–83.CrossRef Spangehl MJ, Masri BA, O’Connell JX, Duncan CP. Prospective analysis of preoperative and intraoperative investigations for the diagnosis of infection at the sites of two hundred and two revision total hip arthroplasties. J Bone Joint Surg Am. 1999;81(5):672–83.CrossRef
15.
go back to reference Zimmerli W, Trampuz A, Ochsner PE. Prosthetic-joint infections. N Engl J Med. 2004;351(16):1645–54.CrossRef Zimmerli W, Trampuz A, Ochsner PE. Prosthetic-joint infections. N Engl J Med. 2004;351(16):1645–54.CrossRef
16.
go back to reference Di Bonaventura G, D'Antonio D, Catamo G, Ballone E, Piccolomini R. Comparison of Etest, agar dilution, broth microdilution and disk diffusion methods for testing in vitro activity of levofloxacin against Staphylococcus spp. isolated from neutropenic cancer patients. Int J Antimicrob Agents. 2002;19(2):147–54.CrossRef Di Bonaventura G, D'Antonio D, Catamo G, Ballone E, Piccolomini R. Comparison of Etest, agar dilution, broth microdilution and disk diffusion methods for testing in vitro activity of levofloxacin against Staphylococcus spp. isolated from neutropenic cancer patients. Int J Antimicrob Agents. 2002;19(2):147–54.CrossRef
17.
go back to reference Lee CK, Barbier BA, Bottos EM, McDonald IR, Cary SC. The Inter-Valley Soil Comparative Survey: the ecology of Dry Valley edaphic microbial communities. ISME J. 2012;6(5):1046–57.CrossRef Lee CK, Barbier BA, Bottos EM, McDonald IR, Cary SC. The Inter-Valley Soil Comparative Survey: the ecology of Dry Valley edaphic microbial communities. ISME J. 2012;6(5):1046–57.CrossRef
18.
go back to reference Goda A, Maruyama F, Michi Y, Nakagawa I, Harada K. Analysis of the factors affecting the formation of the microbiome associated with chronic osteomyelitis of the jaw. Clin Microbiol Infect. 2014;20(5):O309–17.CrossRef Goda A, Maruyama F, Michi Y, Nakagawa I, Harada K. Analysis of the factors affecting the formation of the microbiome associated with chronic osteomyelitis of the jaw. Clin Microbiol Infect. 2014;20(5):O309–17.CrossRef
19.
go back to reference Quast C, Pruesse E, Yilmaz P, et al. The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res. 2013;41(Database issue):D590–6.PubMed Quast C, Pruesse E, Yilmaz P, et al. The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res. 2013;41(Database issue):D590–6.PubMed
20.
go back to reference Edgar RC. Search and clustering orders of magnitude faster than BLAST. Bioinformatics. 2010;26(19):2460–1.CrossRef Edgar RC. Search and clustering orders of magnitude faster than BLAST. Bioinformatics. 2010;26(19):2460–1.CrossRef
21.
go back to reference Reizner W, Hunter JG, O'Malley NT, Southgate RD, Schwarz EM, Kates SL. A systematic review of animal models for Staphylococcus aureus osteomyelitis. Eur Cell Mater. 2014;27:196–212.CrossRef Reizner W, Hunter JG, O'Malley NT, Southgate RD, Schwarz EM, Kates SL. A systematic review of animal models for Staphylococcus aureus osteomyelitis. Eur Cell Mater. 2014;27:196–212.CrossRef
22.
go back to reference Isigidi BK, Devriese LA, Van Hoof J. A note on the isolation of Staphylococcus aureus from raw minced meat. J Appl Bacteriol. 1985;59(5):403–6.CrossRef Isigidi BK, Devriese LA, Van Hoof J. A note on the isolation of Staphylococcus aureus from raw minced meat. J Appl Bacteriol. 1985;59(5):403–6.CrossRef
23.
go back to reference Lin MH, Shu JC, Lin LP, et al. Elucidating the crucial role of poly N-acetylglucosamine from Staphylococcus aureus in cellular adhesion and pathogenesis. PLoS One. 2015;10(4):e0124216.CrossRef Lin MH, Shu JC, Lin LP, et al. Elucidating the crucial role of poly N-acetylglucosamine from Staphylococcus aureus in cellular adhesion and pathogenesis. PLoS One. 2015;10(4):e0124216.CrossRef
24.
go back to reference Bustin SA, Benes V, Garson JA, et al. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009;55(4):611–22.CrossRef Bustin SA, Benes V, Garson JA, et al. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009;55(4):611–22.CrossRef
25.
go back to reference Iijima M, Muguruma T, Brantley W, et al. Effect of coating on properties of esthetic orthodontic nickel-titanium wires. Angle Orthod. 2012;82(2):319–25.CrossRef Iijima M, Muguruma T, Brantley W, et al. Effect of coating on properties of esthetic orthodontic nickel-titanium wires. Angle Orthod. 2012;82(2):319–25.CrossRef
26.
go back to reference Ivanov IE, Kintz EN, Porter LA, et al. Relating the physical properties of Pseudomonas aeruginosa lipopolysaccharides to virulence by atomic force microscopy. J Bacteriol. 2011;193(5):1259–66.CrossRef Ivanov IE, Kintz EN, Porter LA, et al. Relating the physical properties of Pseudomonas aeruginosa lipopolysaccharides to virulence by atomic force microscopy. J Bacteriol. 2011;193(5):1259–66.CrossRef
27.
go back to reference Chen J, Li T, Zhou X, et al. Characterization of the clustered regularly interspaced short palindromic repeats sites in Streptococcus mutans isolated from early childhood caries patients. Arch Oral Biol. 2017;83:174–80.CrossRef Chen J, Li T, Zhou X, et al. Characterization of the clustered regularly interspaced short palindromic repeats sites in Streptococcus mutans isolated from early childhood caries patients. Arch Oral Biol. 2017;83:174–80.CrossRef
28.
go back to reference Sedghizadeh PP, Kumar SK, Gorur A, et al. Microbial biofilms in osteomyelitis of the jaw and osteonecrosis of the jaw secondary to bisphosphonate therapy. J Am Dent Assoc. 2009;140(10):1259–65.CrossRef Sedghizadeh PP, Kumar SK, Gorur A, et al. Microbial biofilms in osteomyelitis of the jaw and osteonecrosis of the jaw secondary to bisphosphonate therapy. J Am Dent Assoc. 2009;140(10):1259–65.CrossRef
29.
go back to reference Gaetti-Jardim E Jr, Landucci LF, de Oliveira KL, et al. Microbiota associated with infections of the jaws. Int J Dent. 2012;2012:369751.CrossRef Gaetti-Jardim E Jr, Landucci LF, de Oliveira KL, et al. Microbiota associated with infections of the jaws. Int J Dent. 2012;2012:369751.CrossRef
30.
go back to reference Cameron DR, Jiang JH, Kostoulias X, Foxwell DJ, Peleg AY. Vancomycin susceptibility in methicillin-resistant Staphylococcus aureus is mediated by YycHI activation of the WalRK essential two-component regulatory system. Sci Rep. 2016;6:30823.CrossRef Cameron DR, Jiang JH, Kostoulias X, Foxwell DJ, Peleg AY. Vancomycin susceptibility in methicillin-resistant Staphylococcus aureus is mediated by YycHI activation of the WalRK essential two-component regulatory system. Sci Rep. 2016;6:30823.CrossRef
31.
go back to reference Zhao Y, Verma V, Belcheva A, et al. Staphylococcus aureus methicillin-resistance factor fmtA is regulated by the global regulator SarA. PLoS One. 2012;7(8):e43998.CrossRef Zhao Y, Verma V, Belcheva A, et al. Staphylococcus aureus methicillin-resistance factor fmtA is regulated by the global regulator SarA. PLoS One. 2012;7(8):e43998.CrossRef
32.
go back to reference Fukuchi K, Kasahara Y, Asai K, et al. The essential two-component regulatory system encoded by yycF and yycG modulates expression of the ftsAZ operon in Bacillus subtilis. Microbiology. 2000;146(Pt 7):1573–83.CrossRef Fukuchi K, Kasahara Y, Asai K, et al. The essential two-component regulatory system encoded by yycF and yycG modulates expression of the ftsAZ operon in Bacillus subtilis. Microbiology. 2000;146(Pt 7):1573–83.CrossRef
33.
go back to reference Martin PK, Li T, Sun D, Biek DP, Schmid MB. Role in cell permeability of an essential two-component system in Staphylococcus aureus. J Bacteriol. 1999;181(12):3666–73.PubMedPubMedCentral Martin PK, Li T, Sun D, Biek DP, Schmid MB. Role in cell permeability of an essential two-component system in Staphylococcus aureus. J Bacteriol. 1999;181(12):3666–73.PubMedPubMedCentral
34.
go back to reference Mascher T, Helmann JD, Unden G. Stimulus perception in bacterial signal-transducing histidine kinases. Microbiol Mol Biol Rev. 2006;70(4):910–38.CrossRef Mascher T, Helmann JD, Unden G. Stimulus perception in bacterial signal-transducing histidine kinases. Microbiol Mol Biol Rev. 2006;70(4):910–38.CrossRef
35.
go back to reference Stein T, Borchert S, Kiesau P, et al. Dual control of subtilin biosynthesis and immunity in Bacillus subtilis. Mol Microbiol. 2002;44(2):403–16.CrossRef Stein T, Borchert S, Kiesau P, et al. Dual control of subtilin biosynthesis and immunity in Bacillus subtilis. Mol Microbiol. 2002;44(2):403–16.CrossRef
36.
go back to reference Dubrac S, Boneca IG, Poupel O, Msadek T. New insights into the WalK/WalR (YycG/YycF) essential signal transduction pathway reveal a major role in controlling cell wall metabolism and biofilm formation in Staphylococcus aureus. J Bacteriol. 2007;189(22):8257–69.CrossRef Dubrac S, Boneca IG, Poupel O, Msadek T. New insights into the WalK/WalR (YycG/YycF) essential signal transduction pathway reveal a major role in controlling cell wall metabolism and biofilm formation in Staphylococcus aureus. J Bacteriol. 2007;189(22):8257–69.CrossRef
37.
go back to reference Howden BP, McEvoy CR, Allen DL, et al. Evolution of multidrug resistance during Staphylococcus aureus infection involves mutation of the essential two component regulator WalKR. PLoS Pathog. 2011;7(11):e1002359.CrossRef Howden BP, McEvoy CR, Allen DL, et al. Evolution of multidrug resistance during Staphylococcus aureus infection involves mutation of the essential two component regulator WalKR. PLoS Pathog. 2011;7(11):e1002359.CrossRef
38.
go back to reference Gardete S, Kim C, Hartmann BM, et al. Genetic pathway in acquisition and loss of vancomycin resistance in a methicillin resistant Staphylococcus aureus (MRSA) strain of clonal type USA300. PLoS Pathog. 2012;8(2):e1002505.CrossRef Gardete S, Kim C, Hartmann BM, et al. Genetic pathway in acquisition and loss of vancomycin resistance in a methicillin resistant Staphylococcus aureus (MRSA) strain of clonal type USA300. PLoS Pathog. 2012;8(2):e1002505.CrossRef
39.
go back to reference Clarke SR, Foster SJ. Surface adhesins of Staphylococcus aureus. Adv Microb Physiol. 2006;51:187–224.CrossRef Clarke SR, Foster SJ. Surface adhesins of Staphylococcus aureus. Adv Microb Physiol. 2006;51:187–224.CrossRef
40.
go back to reference Bazari PAM, Honarmand Jahromy S, Zare Karizi S. Phenotypic and genotypic characterization of biofilm formation among Staphylococcus aureus isolates from clinical specimens, an atomic force microscopic (AFM) study. Microb Pathog. 2017;110:533–9. Bazari PAM, Honarmand Jahromy S, Zare Karizi S. Phenotypic and genotypic characterization of biofilm formation among Staphylococcus aureus isolates from clinical specimens, an atomic force microscopic (AFM) study. Microb Pathog. 2017;110:533–9.
Metadata
Title
Staphylococcus aureus biofilm organization modulated by YycFG two-component regulatory pathway
Authors
Shizhou Wu
Fuguo Huang
Hui Zhang
Lei Lei
Publication date
01-12-2019
Publisher
BioMed Central
Keyword
Osteomyelitis
Published in
Journal of Orthopaedic Surgery and Research / Issue 1/2019
Electronic ISSN: 1749-799X
DOI
https://doi.org/10.1186/s13018-018-1055-z

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