BIOFILM AND ROLE OF icaA, icaD, AND ACCUMULATION-ASSOCIATED PROTEIN IN STAPHYLOCOCCUS EPIDERMIDIS ISOLATED IN BREAST SURGERY_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

TANGQi ¹ , YUANBing ² ,  HUANGYunchao ² , GUOFengli ³ , LEIYujie ²

1. Department of Breast Surgery, the Third Affiliated Hospital of Kunming Medical University Kunming Yunnan, 650118, P. R. China;
2. Department of Cardiovascular Thoracic Surgery, the Third Affiliated Hospital of Kunming Medical University Kunming Yunnan, 650118, P. R. China;
3. Department of Clinical Laboratory, the Third Affiliated Hospital of Kunming Medical University, Kunming Yunnan, 650118, P. R. China;

Corresponding author：

HUANGYunchao, Email: huangych2001@aliyun.com

Keywords：

Breast surgery; Staphylococcus epidermidis; Bacterial biofilm; icaA; icaD; Accumulation-associated protein

DOI：

10.7507/1002-1892.20140053

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Objective To investigate biofilm formation on the surface of silica gel by breast surgery clinical specimens of Staphylococcus epidermidis and to analyze the relationship between biofilm formation and icaA, icaD, and accumulation^-associated protein (aap) gene. Methods Between December 2011 and January 2013, 44 strains of Staphylococcus epidermidis were isolated from the clinical specimens of the female patients who had no symptom of infection. The icaA, icaD, and aap genes were detected by PCR and 4 genotypic groups were divided:icaA⁺icaD⁺/aap⁺ group (group A), icaA⁺icaD⁺/aap^- group (group B), icaA^-icaD^-/aap⁺ group (group C), and icaA^-icaD^-/aap^- group (group D). Biofilms mass was semi^-quantified by semi^-quantitative adherence assay after 8, 12, 24, 30, and 36 hours of incubation. The thickness of biofilms was measured by confocal laser scanning microscope (CLSM) at 12 and 24 hours after incubation. The ultrastructure of biofilms was observed by scanning electron microscope (SEM) at 24 hours after incubation. Results PCR test showed that 13 strains were icaA⁺icaD⁺/aap⁺(group A), 12 strains were icaA⁺icaD⁺/aap^-(group B), 16 strains were icaA^-icaD^-/aap⁺(group C), and 3 strains were icaA^-icaD^-/aap^-(group D). In 29 strains which had bacterial biofilm formation (65.9%), there were 13 strains in group A, 7 strains in group B, 9 strains in group C, and 0 in group D. The result of semi^-quantitative adherence assay showed no significant difference in the absorbance (A) values among 4 groups at 8 hours (P>0.05). The A values of groups A, B, and C were significantly higher than that of group D at 12^-36 hours, and group A was significantly higher than groups B and C (P<0.05), but there was no significant difference between groups B and C (P>0.05). The results of CLSM showed that the thickness of biofilm in groups A, B, and C was significantly larger than that in group D at 12 and 24 hours after incubation (P<0.05), and the thickness of biofilm in group A was significantly larger than that in groups B and C (P<0.05), but there was no significant difference between groups B and C (P>0.05). The result of SEM showed that the mature biofilm could be observed on the surface of silica gel in groups A, B, and C, and the ultrastructure of biofilms in group A were the most abundant and extensive among 3 groups. The ultrastructure of biofilm in group B was similar to that in group C. No obvious biofilms formed in group D. Conclusion icaA, icaD, and aap genes all play key roles in the process for biofilm formation of Staphylococcus epidermidis. Futhermore, aap gene enhance the ability of biofilm^-forming when aap and ica genes coexist, so the biofilm^-forming ability of icaA⁺icaD⁺/aap⁺ is strongest.

Citation： TANGQi, YUANBing, HUANGYunchao, GUOFengli, LEIYujie. BIOFILM AND ROLE OF icaA, icaD, AND ACCUMULATION-ASSOCIATED PROTEIN IN STAPHYLOCOCCUS EPIDERMIDIS ISOLATED IN BREAST SURGERY. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(2): 244-249. doi: 10.7507/1002-1892.20140053 Copy

1.	Jacombs A,Allan J,Hu H,et al.Prevention of biofilm-induced capsular contracture with antibiotic-impregnated mesh in a porcine model.Aesthet Surg J,2012,32(7):886-891.
2.	Handel N,Cordray T,Gutierrez J,et al.A long-term study of outcomes,complications,and patients satisfactions with breast implants.Plast Reconstr Surg,2006,117(3):757-772.
3.	Wixtrom RN,Stutman RL,Burke RM,et al.Risk of breast implant bacterial contamination from endogenous breast flora,prevention with nipple shields,and implications for biofilm formation.Aesthet Surg,2012,32(8):956-963.
4.	Nuryastuti T,Krom BP,Aman AT,et al.Ica-expression and gentamicin susceptibility of Staphylococcus epidermidis biofilm on orthopedic implant biomaterials.J Biomed Mater Res A,2011,96(2):365-371.
5.	叶联华,黄云超,李高峰,等.生物材料植入后发生感染与表皮葡萄球菌生物膜.生物医学工程与临床,2010,14(1):79-83.
6.	Parsek MR,Singh PK.Bacterial biofilms:an emerging link to disease pathogenesis.Annu Rev Microbiol,2003,57:677-701.
7.	叶联华,黄云超,杨达宽,等.葡萄糖对胸心外科聚氯乙烯材料细菌ica操纵子表型的影响.中华医院感染学杂志,2007,17(5):481-484.
8.	叶联华,黄云超,许赓,等.医源性表皮葡萄球菌胞间黏附素基因操纵子在聚氯乙烯材料表面细菌生物膜形成中的作用研究.中国修复重建外科杂志,2011,25(4):466-471.
9.	Agarwal A,Jain A.Glucose&sodium chloride induced biofilm production&ica operon in clinical isolates of Staphylococci.Indian Journal of Medical Research,2013,138(2):262-266.
10.	Kadry AA,Fouda SI,Shibl AM,et al.Impact of slime dispersants and anti-adhesives on in vitro biofilm formation of Staphylococcus epidermidis on intraocular lenses and on antibiotic activities.J Antimicrob Chemother,2009,63(3):480-484.
11.	Arciola CR,Campoccia D,Baldassarri L,et al.Detection of biofilm formation in Staphylococcus epidermidis from implant infections.Comparison of a PCR-method that recognizes the presence of ica genes with two classic phenotypic methods.J Biomed Mater Res A,2006,76(2):425-430.
12.	雷玉洁,黄云超,杨丽,等.肺癌患者中心静脉导管相关表皮葡萄球菌icaA、icaD表达及转化生长因子β₁与生物膜的关系.中国组织工程研究,2012,16(12):2158-2162.
13.	Banner MA,Cunniffe JG,Macintosh RL,et al.Localized tufts of fibrils on Staphylococcus epidermidis NCTC 11047 are comprised of the accumulation-associated protein.J Bacteriolo,2007,189(7):2793-2804.
14.	Rohde H,Burandt EC,Siemssen N,et al.Polysaccharide intercellular adhesion or protein factorsin biofilm accumulation of Staphylococcus epidermidis and Staphylococcus aureus isolated from prosthetichip and knee joint infections.Biomaterials,2007,28(9):1711-1720.
15.	Macintosh RL,Brittan JL,Bhattacharya R,et al.The terminal A domain of the fibrillar accumulation-associated protein (Aap) of Staphylococcus epidermidis mediates adhesion to human corneocytes.J Bacteriol,2009,191(22):7007-7016.
16.	Hu J,Xu T,Zhu T,et al.Monoclonal antibodies against accumulation-associated protein affect EPS biosynthesis and enhance bacterial accumulation of Staphylococcus epidermidis.PLoS One,2011,6(6):e20918.
17.	Golus J,Stankevic M,Sawicki R,et al.Quantitative analysis of biofilm formed on vascular prostheses by Staphylococcus epidermidis with different ica and aap genetic status.Int J Artif Organs,2013,36(2):105-112.
18.	Conrady DG,Brescia CC,Horii K,et al.A zinc-dependent adhesion module is responsible for intercellular adhesion in staphylococcal biofilms.Proc Natl Acad Sci U S A,2008,105(49):19456-19461.
19.	Bateman A,Holden MT,Yeats C.The G5 domain:a potential N-acetylglucosamine recognition domain in biofilm formation.Bioinformatics,2005,21(8):1301-1303.
20.	Stevens NT,Tharmabala M,Dillane T,et al.Biofilmand the role of theica operon and aap in Staphylococcus epidermidis isolates causing neurosurgical meningitis.Clin Microbiol Infect,2008,14(7):719-722.

1. Jacombs A,Allan J,Hu H,et al.Prevention of biofilm-induced capsular contracture with antibiotic-impregnated mesh in a porcine model.Aesthet Surg J,2012,32(7):886-891.
2. Handel N,Cordray T,Gutierrez J,et al.A long-term study of outcomes,complications,and patients satisfactions with breast implants.Plast Reconstr Surg,2006,117(3):757-772.
3. Wixtrom RN,Stutman RL,Burke RM,et al.Risk of breast implant bacterial contamination from endogenous breast flora,prevention with nipple shields,and implications for biofilm formation.Aesthet Surg,2012,32(8):956-963.
4. Nuryastuti T,Krom BP,Aman AT,et al.Ica-expression and gentamicin susceptibility of Staphylococcus epidermidis biofilm on orthopedic implant biomaterials.J Biomed Mater Res A,2011,96(2):365-371.
5. 叶联华,黄云超,李高峰,等.生物材料植入后发生感染与表皮葡萄球菌生物膜.生物医学工程与临床,2010,14(1):79-83.
6. Parsek MR,Singh PK.Bacterial biofilms:an emerging link to disease pathogenesis.Annu Rev Microbiol,2003,57:677-701.
7. 叶联华,黄云超,杨达宽,等.葡萄糖对胸心外科聚氯乙烯材料细菌ica操纵子表型的影响.中华医院感染学杂志,2007,17(5):481-484.
8. 叶联华,黄云超,许赓,等.医源性表皮葡萄球菌胞间黏附素基因操纵子在聚氯乙烯材料表面细菌生物膜形成中的作用研究.中国修复重建外科杂志,2011,25(4):466-471.
9. Agarwal A,Jain A.Glucose&sodium chloride induced biofilm production&ica operon in clinical isolates of Staphylococci.Indian Journal of Medical Research,2013,138(2):262-266.
10. Kadry AA,Fouda SI,Shibl AM,et al.Impact of slime dispersants and anti-adhesives on in vitro biofilm formation of Staphylococcus epidermidis on intraocular lenses and on antibiotic activities.J Antimicrob Chemother,2009,63(3):480-484.
11. Arciola CR,Campoccia D,Baldassarri L,et al.Detection of biofilm formation in Staphylococcus epidermidis from implant infections.Comparison of a PCR-method that recognizes the presence of ica genes with two classic phenotypic methods.J Biomed Mater Res A,2006,76(2):425-430.
12. 雷玉洁,黄云超,杨丽,等.肺癌患者中心静脉导管相关表皮葡萄球菌icaA、icaD表达及转化生长因子β₁与生物膜的关系.中国组织工程研究,2012,16(12):2158-2162.
13. Banner MA,Cunniffe JG,Macintosh RL,et al.Localized tufts of fibrils on Staphylococcus epidermidis NCTC 11047 are comprised of the accumulation-associated protein.J Bacteriolo,2007,189(7):2793-2804.
14. Rohde H,Burandt EC,Siemssen N,et al.Polysaccharide intercellular adhesion or protein factorsin biofilm accumulation of Staphylococcus epidermidis and Staphylococcus aureus isolated from prosthetichip and knee joint infections.Biomaterials,2007,28(9):1711-1720.
15. Macintosh RL,Brittan JL,Bhattacharya R,et al.The terminal A domain of the fibrillar accumulation-associated protein (Aap) of Staphylococcus epidermidis mediates adhesion to human corneocytes.J Bacteriol,2009,191(22):7007-7016.
16. Hu J,Xu T,Zhu T,et al.Monoclonal antibodies against accumulation-associated protein affect EPS biosynthesis and enhance bacterial accumulation of Staphylococcus epidermidis.PLoS One,2011,6(6):e20918.
17. Golus J,Stankevic M,Sawicki R,et al.Quantitative analysis of biofilm formed on vascular prostheses by Staphylococcus epidermidis with different ica and aap genetic status.Int J Artif Organs,2013,36(2):105-112.
18. Conrady DG,Brescia CC,Horii K,et al.A zinc-dependent adhesion module is responsible for intercellular adhesion in staphylococcal biofilms.Proc Natl Acad Sci U S A,2008,105(49):19456-19461.
19. Bateman A,Holden MT,Yeats C.The G5 domain:a potential N-acetylglucosamine recognition domain in biofilm formation.Bioinformatics,2005,21(8):1301-1303.
20. Stevens NT,Tharmabala M,Dillane T,et al.Biofilmand the role of theica operon and aap in Staphylococcus epidermidis isolates causing neurosurgical meningitis.Clin Microbiol Infect,2008,14(7):719-722.

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BIOFILM AND ROLE OF icaA, icaD, AND ACCUMULATION-ASSOCIATED PROTEIN IN STAPHYLOCOCCUS EPIDERMIDIS ISOLATED IN BREAST SURGERY

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