Effect of acidic and alkali shocks on expression of efaA gene in Enterococcus faecalis, isolated from root canal infection

Mahsa Beomidehagh; Mohammad Ahangarzadeh Rezaee; Khudaverdi Ganbarov; Farnaz Jafari; Alka Hasani; Naser Alizadeh; Asghar Tanomand; Hossein Samadi Kafil

doi:10.14715/cmb/2018.64.13.1

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Vol. 64 No. 13: Issue 13

Issue Published : November 2, 2018

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Effect of acidic and alkali shocks on expression of efaA gene in Enterococcus faecalis, isolated from root canal infection

https://doi.org/10.14715/cmb/2018.64.13.1

Mahsa Beomidehagh

Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran

Mohammad Ahangarzadeh Rezaee

Immunology Research Center, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

Khudaverdi Ganbarov

Department of Microbiology, Baku State University, Baku, Republic of Azerbaijan

Farnaz Jafari

Department of Endodontics, Dental School, Tabriz University of Medical Sciences, Tabriz, Iran

Alka Hasani

Immunology Research Center, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

Naser Alizadeh

Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran

Asghar Tanomand

Department of Basic Sciences, Maragheh University of Medical Sciences, Maragheh, Iran

Hossein Samadi Kafil

Immunology Research Center, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

Corresponding Author(s) : Hossein Samadi Kafil

Kafilhs@tbzmed.ac.ir

Cellular and Molecular Biology, Vol. 64 No. 13: Issue 13
Article Published : October 30, 2018

Abstract

Enterococcus faecalis is an important opportunistic infectious agent involving the oral cavity and endodontics. The aim of this study was to evaluate the expression ratio of efaA gene in biofilm producer E. faecalis before and after receiving acidic and alkali shocks. One hundred E. faecalis isolates were gathered from 170 infectious root canals. After analysis of biofilm formation by the Microtiterplate method, the presence of efaA gene was examined by PCR and its expression was evaluated by Real-time PCR, one before applying any stressed to isolates and another by applying acidic and alkali shock. Chi-square method was used for statistical analysis. Eighty-two percent of samples had efaA gene. Evaluation of biofilm formation, 49% of the isolates were strong biofilm producer, 42% moderate and 10 % of them had no biofilm. 59% overexpression of efaA gene was observed in biofilm producer isolates, while there were no significant changes in samples with acidic stress and decreased expression after alkali shock. Findings of the present study, indicates importance of efaA gene in biofilm formation and pathogenesis of E. faecalis. Acid had no effect of expression of this gene but alkali reduced expression of this gene in a significant level. These results indicate the importance of efaA and acidic conditions in biofilm production by E. faecalis.

Keywords

Acid response Adhesin Biofilms Gene expression Endodontic infection.

Beomidehagh, M., Rezaee, M. A., Ganbarov, K., Jafari, F., Hasani, A., Alizadeh, N., Tanomand, A., & Kafil, H. S. (2018). Effect of acidic and alkali shocks on expression of efaA gene in Enterococcus faecalis, isolated from root canal infection. Cellular and Molecular Biology, 64(13), 1–5. https://doi.org/10.14715/cmb/2018.64.13.1

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References

References
Teymournejad O, Mohabati Mobarez A, Hosseini Doust R. Epidemiologic evaluation of Vancomycin Resistant genes in Enterococcus spp. isolated from clinical samples. Journal of Fasa University of Medical Sciences. 2011;1(2):1-6.
Kafil HS, Mobarez AM. Spread of Enterococcal Surface Protein in Antibiotic Resistant Entero-coccus faecium and Enterococcus faecalis isolates from Urinary Tract Infections. Open Microbiol J. 2015;9:14-7.
Akbari Aghdam M, Soroush Barhaghi MH, Aghazadeh M, Jafari F, Beomide Hagh M, Haghdoost M, et al. Virulence genes in biofilm producer Enterococcus faecalis isolates from root canal infections. Cell Mol Biol. 2017;63(5):55-9.
Kafil HS, Mobarez AM, Moghadam MF. Adhesion and virulence factor properties of Enterococci isolated from clinical samples in Iran. Indian Journal of Pathology and Microbiology. 2013;56(3):238.
Aas JA, Paster BJ, Stokes LN, Olsen I, Dewhirst FE. Defining the normal bacterial flora of the oral cavity. Journal of clinical microbiology. 2005;43(11):5721-32.
Dahlen G, Samuelsson W, Molander A, Reit C. Identification and antimicrobial susceptibility of enterococci isolated from the root canal. Oral microbiology and Immunology. 2000;15(5):309-12.
Portenier I, Waltimo TM, Haapasalo M. Enterococcus faecalis–the root canal survivor and ‘star'in post"treatment disease. Endodontic topics. 2003;6(1):135-59.
Kafil HS, Mobarez AM. Assessment of biofilm formation by enterococci isolates from urinary tract infections with different virulence profiles. Journal of King Saud University-Science. 2015;27(4):312-7.
Kafil HS, Asgharzadeh M. Vancomycin-resistant enteroccus faecium and enterococcus faecalis isolated from education hospital of iran. Maedica. 2014;9(4):323-7.
Kafil HS, Mobarez AM, Moghadam MF, Hashemi ZS, Yousefi M. Gentamicin induces efaA expression and biofilm formation in Enterococcus faecalis. Microb Pathog. 2016;92:30-5.
Fattahi S, Aghazadeh M, Nahaei MR, Asgharzadeh M, Kafil HS. Comparison of virulence factors fimA, papC, and hly among uropathogenic Escherichia coli isolates producing and nonproducing extended spectrum beta-lactamases. Annals of Tropical Medicine and Public Health. 2017;10(2):404.
Creti R, Koch S, Fabretti F, Baldassarri L, Huebner J. Enterococcal colonization of the gastro-intestinal tract: role of biofilm and environmental oligosaccharides. BMC microbiology. 2006;6(1):60.
Mohamed JA, Huang W, Nallapareddy SR, Teng F, Murray BE. Influence of origin of isolates, especially endocarditis isolates, and various genes on biofilm formation by Enterococcus faecalis. Infection and immunity. 2004;72(6):3658-63.
Rich RL, Kreikemeyer B, Owens RT, LaBrenz S, Narayana SV, Weinstock GM, et al. Ace is a collagen-binding MSCRAMM from Enterococcus faecalis. Journal of Biological Chemistry. 1999;274(38):26939-45.
Smyth C, MATTHEWS H, Halpenny M, Brandis H, Colman G. Biotyping, serotyping and phage typing of Streptococcus faecalis isolated from dental plaque in the human mouth. Journal of medical microbiology. 1987;23(1):45-54.
Sánchez CJ, Mende K, Beckius ML, Akers KS, Romano DR, Wenke JC, et al. Biofilm formation by clinical isolates and the implications in chronic infections. BMC infectious diseases. 2013;13(1):47.
Hatt JK, Rather PN. Role of bacterial biofilms in urinary tract infections. Current topics in microbiology and immunology. 2008;322:163-92.
Jabbari V, Mokarram RR, Khiabani MS, Askari F, Ahmadi E, mohammad Hassanzadeh A, et al. Molecular Identification of Lactobacillus acidophilus as a probiotic potential from traditional doogh samples and evaluation of their antimicrobial activity against some pathogenic bacteria. Biomedical Research. 2017;28(2):0970-938X.
Asgharzadeh M, Kafil HS, Ebrahimzadeh ME, A B. Mannose-Binding Lectin Gene and Promoter Polymorphism and Susceptibility to Renal Dysfunction in Systemic Lupus Erythematosus. Journal of Biological Sciences. 2007;7(5):801-5.
Bore E, Langsrud S, Langsrud í˜, Rode TM, Holck A. Acid-shock responses in Staphylococcus aureus investigated by global gene expression analysis. Microbiology. 2007;153(7):2289-303.
Asgharzadeh M, Shahbabian K, Kafil HS, Rafi A. Use of DNA Fingerprinting in Identifying the Source Case of Tuberculosis in East Azarbaijan Province of Iran. J Med Sci. 2007;7(3):418-21.
Hancock H, Sigurdsson A, Trope M, Moiseiwitsch J. Bacteria isolated after unsuccessful endodontic treatment in a North American population. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology. 2001;91(5):579-86.
Peciuliene V, Reynaud A, Balciuniene I, Haapasalo M. Isolation of yeasts and enteric bacteria in root"filled teeth with chronic apical periodontitis. International endodontic journal. 2001;34(6):429-34.
Flahaut S, Hartke A, Giard J-C, Auffray Y. Alkaline stress response in Enterococcus faecalis: adaptation, cross-protection, and changes in protein synthesis. Applied and environmental microbiology. 1997;63(2):812-4.
Shankar N, Lockatell CV, Baghdayan AS, Drachenberg C, Gilmore MS, Johnson DE. Role of Enterococcus faecalissurface protein ESP in the pathogenesis of ascending urinary tract infection. Infection and immunity. 2001;69(7):4366-72.
Preethee T, Kandaswamy D, Hannah R. Molecular identification of an Enterococcus faecalis endocarditis antigen efaA in root canals of therapy-resistant endodontic infections. Journal of Conservative Dentistry : JCD. 2012 Oct-Dec;15(4):319-22.
Zoletti GO, Pereira EM, Schuenck RP, Teixeira LM, Siqueira JF, dos Santos KRN. Characterization of virulence factors and clonal diversity of Enterococcus faecalis isolates from treated dental root canals. Research in Microbiology. 2011 2011/02/01/;162(2):151-8.

References

Teymournejad O, Mohabati Mobarez A, Hosseini Doust R. Epidemiologic evaluation of Vancomycin Resistant genes in Enterococcus spp. isolated from clinical samples. Journal of Fasa University of Medical Sciences. 2011;1(2):1-6.

Kafil HS, Mobarez AM. Spread of Enterococcal Surface Protein in Antibiotic Resistant Entero-coccus faecium and Enterococcus faecalis isolates from Urinary Tract Infections. Open Microbiol J. 2015;9:14-7.

Akbari Aghdam M, Soroush Barhaghi MH, Aghazadeh M, Jafari F, Beomide Hagh M, Haghdoost M, et al. Virulence genes in biofilm producer Enterococcus faecalis isolates from root canal infections. Cell Mol Biol. 2017;63(5):55-9.

Kafil HS, Mobarez AM, Moghadam MF. Adhesion and virulence factor properties of Enterococci isolated from clinical samples in Iran. Indian Journal of Pathology and Microbiology. 2013;56(3):238.

Aas JA, Paster BJ, Stokes LN, Olsen I, Dewhirst FE. Defining the normal bacterial flora of the oral cavity. Journal of clinical microbiology. 2005;43(11):5721-32.

Dahlen G, Samuelsson W, Molander A, Reit C. Identification and antimicrobial susceptibility of enterococci isolated from the root canal. Oral microbiology and Immunology. 2000;15(5):309-12.

Portenier I, Waltimo TM, Haapasalo M. Enterococcus faecalis–the root canal survivor and ‘star'in post"treatment disease. Endodontic topics. 2003;6(1):135-59.

Kafil HS, Mobarez AM. Assessment of biofilm formation by enterococci isolates from urinary tract infections with different virulence profiles. Journal of King Saud University-Science. 2015;27(4):312-7.

Kafil HS, Asgharzadeh M. Vancomycin-resistant enteroccus faecium and enterococcus faecalis isolated from education hospital of iran. Maedica. 2014;9(4):323-7.

Kafil HS, Mobarez AM, Moghadam MF, Hashemi ZS, Yousefi M. Gentamicin induces efaA expression and biofilm formation in Enterococcus faecalis. Microb Pathog. 2016;92:30-5.

Fattahi S, Aghazadeh M, Nahaei MR, Asgharzadeh M, Kafil HS. Comparison of virulence factors fimA, papC, and hly among uropathogenic Escherichia coli isolates producing and nonproducing extended spectrum beta-lactamases. Annals of Tropical Medicine and Public Health. 2017;10(2):404.

Creti R, Koch S, Fabretti F, Baldassarri L, Huebner J. Enterococcal colonization of the gastro-intestinal tract: role of biofilm and environmental oligosaccharides. BMC microbiology. 2006;6(1):60.

Mohamed JA, Huang W, Nallapareddy SR, Teng F, Murray BE. Influence of origin of isolates, especially endocarditis isolates, and various genes on biofilm formation by Enterococcus faecalis. Infection and immunity. 2004;72(6):3658-63.

Rich RL, Kreikemeyer B, Owens RT, LaBrenz S, Narayana SV, Weinstock GM, et al. Ace is a collagen-binding MSCRAMM from Enterococcus faecalis. Journal of Biological Chemistry. 1999;274(38):26939-45.

Smyth C, MATTHEWS H, Halpenny M, Brandis H, Colman G. Biotyping, serotyping and phage typing of Streptococcus faecalis isolated from dental plaque in the human mouth. Journal of medical microbiology. 1987;23(1):45-54.

Sánchez CJ, Mende K, Beckius ML, Akers KS, Romano DR, Wenke JC, et al. Biofilm formation by clinical isolates and the implications in chronic infections. BMC infectious diseases. 2013;13(1):47.

Hatt JK, Rather PN. Role of bacterial biofilms in urinary tract infections. Current topics in microbiology and immunology. 2008;322:163-92.

Jabbari V, Mokarram RR, Khiabani MS, Askari F, Ahmadi E, mohammad Hassanzadeh A, et al. Molecular Identification of Lactobacillus acidophilus as a probiotic potential from traditional doogh samples and evaluation of their antimicrobial activity against some pathogenic bacteria. Biomedical Research. 2017;28(2):0970-938X.

Asgharzadeh M, Kafil HS, Ebrahimzadeh ME, A B. Mannose-Binding Lectin Gene and Promoter Polymorphism and Susceptibility to Renal Dysfunction in Systemic Lupus Erythematosus. Journal of Biological Sciences. 2007;7(5):801-5.

Bore E, Langsrud S, Langsrud í˜, Rode TM, Holck A. Acid-shock responses in Staphylococcus aureus investigated by global gene expression analysis. Microbiology. 2007;153(7):2289-303.

Asgharzadeh M, Shahbabian K, Kafil HS, Rafi A. Use of DNA Fingerprinting in Identifying the Source Case of Tuberculosis in East Azarbaijan Province of Iran. J Med Sci. 2007;7(3):418-21.

Hancock H, Sigurdsson A, Trope M, Moiseiwitsch J. Bacteria isolated after unsuccessful endodontic treatment in a North American population. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology. 2001;91(5):579-86.

Peciuliene V, Reynaud A, Balciuniene I, Haapasalo M. Isolation of yeasts and enteric bacteria in root"filled teeth with chronic apical periodontitis. International endodontic journal. 2001;34(6):429-34.

Flahaut S, Hartke A, Giard J-C, Auffray Y. Alkaline stress response in Enterococcus faecalis: adaptation, cross-protection, and changes in protein synthesis. Applied and environmental microbiology. 1997;63(2):812-4.

Shankar N, Lockatell CV, Baghdayan AS, Drachenberg C, Gilmore MS, Johnson DE. Role of Enterococcus faecalissurface protein ESP in the pathogenesis of ascending urinary tract infection. Infection and immunity. 2001;69(7):4366-72.

Preethee T, Kandaswamy D, Hannah R. Molecular identification of an Enterococcus faecalis endocarditis antigen efaA in root canals of therapy-resistant endodontic infections. Journal of Conservative Dentistry : JCD. 2012 Oct-Dec;15(4):319-22.

Zoletti GO, Pereira EM, Schuenck RP, Teixeira LM, Siqueira JF, dos Santos KRN. Characterization of virulence factors and clonal diversity of Enterococcus faecalis isolates from treated dental root canals. Research in Microbiology. 2011 2011/02/01/;162(2):151-8.

Author biographies is not available.