BibTex RIS Cite

Tek ve iki türlü biyofimlerde E.faecalis jelatinaz gen ekspresyon seviyelerinin belirlenmesi

Year 2015, Issue: 2, 162 - 173, 01.06.2015

Abstract

Biyofilmler insan hayatını tehdit eden önemli bir problemdir. Son yıllarda, persistan enfeksiyonlara yol açan biyofilmlerin çoğunun polimikrobiyal olduğu gösterilmiştir. Enterococcus faecalis idrar yolu, yara enfeksiyonları vb. gibi hastane enfeksiyonlarından sıklıkla izole edilen fırsatçı bir patojendir. E.faecalis’in bazı virulans faktörlerinin ekspresyon seviyelerinde, biyofilm oluşturabilen ve oluşturamayan izolatları arasında farklılıklar gözlenmiştir. Jelatinaz, jelatini, kollajeni ve kazeini hidrolize eden metallo-proteaz enzimi olup, E.faecalis’in virulans faktörlerinden birisidir. Mutant suşlar kullanılarak yapılan bazı çalışmalarda bu enzimin biyofilm oluşumundaki rolü bulunmuştur. İlk amacımız, 11 E.faecalis idrar izolatının biyofilm oluşturabilme prevalansını belirlemektir. İkinci amacımız, tekrar edilebilirliği yüksek iki farklı mikrobiyal türden oluşan biyofilm modelini in vitro ortamda geliştirmek ve C.albicans’ın ortamdaki varlığının E.faecalis’in gelE gen ekspresyonu üzerindeki etkisinin kantitatif RT-PCR ile belirleyebilmektir. Çalışmamızda, test edilen her iki yöntemde de tüm izolatların biyofilm oluşturabildiği saptanmıştır. Ayrıca, E.faecalis ATCC 29212 suşu tarafından oluşturulan monomikrobiyal biyofilm modeline oranla, C.albicans ATCC 90028 ve E.faecalis ATCC 29212 tarafından oluşturulan ikili biyofilm modelinde gelE ekspresyon artışı olduğu sonuçlarımızda gösterilmiştir. İki türden oluşan biyofilm modelinde gözlenen bu gelE ekspresyon seviyesindeki artışı desteklemek amacıyla klinik izolatlar kullanılarak yapılacak çalışmalara ihtiyaç duyulmaktadır.

References

  • Choe, H.S., Son, S.W., Choi, H.A., Kim, H.J., Ahn, S.G., Bang, J.H., Lee, S.J., Lee, J.Y., Cho, Y.H., Lee, S.S.: Analysis of the distribution of bacteria within urinary catheter biofilms using four different molecular techniques, Am J Infect Control, 40, e249-254 (2012)
  • Kart, D., Tavernier, S., Van Acker, H., Nelis, H. J., Coenye, T.: Activity of disinfectants against multispecies biofilms formed by Staphylococcus aureus, Candida albicans and Pseudomonas aeruginosa, Biofouling, 30(3) , 377-383 (2014)
  • Yang, L., Liu, Y., Wu, H., Hİiby, N., Molin, S., Song, Z.J.: Current understanding of multi- species biofilms, IJOS, 3, 2, 74 (2011)
  • Wolcott, R., Costerton, J.W., Raoult, D., Cutler, S.J.: The polymicrobial nature of biofilm infection. Clin Microbiol Infect, 19(2), 107-112 (2013)
  • Weimer, K.E., Juneau, R.A., Murrah, K.A. et al.: Divergent mechanisms for passive pneumococcal resistance to beta-lactam antibiotics in the presence of Haemophilus influenzae, J Infect Dis, 203, 549–555 (2011)
  • Elias, S., Banin, E.: Multi-species biofilms: living with friendly neighbors, FEMS Microbiol Rev, Doi: 10.1111/j.1574-6976.2012.00325.x. [Epub ahead of print] (2012)
  • Fischbach, M.A., Sonnenburg, J.L.: Eating for two: how metabolism estab- lishes interspecies interactions in the gut, Cell Host Microbe, 10, 336–347 (2011)
  • Carlsson, J. : Bacterial metabolism in dental biofilms, Adv Dent Res, 11, 75–80 (1997)
  • Madsen, J.S., Burmolle, M., Hansen, L.H., Sorensen, S.J.: The interconnection between biofilm formation and horizontal gene transfer, FEMS Immunol Med Microbiol, 65, 183–195 (2012)
  • Arciola, C.R., Baldassarri, L., Campoccia, D., Creti, R., Prini, V., Huebner, J., Montanaro, L.: Strong biofilm production, antibiotic multi-resistance and high gelE expression in epidemic clones of Enterococcus faecalis from orthopaedic implant infections, Biomaterials, 29, 580- 586 (2008)
  • Ruoff, K.L., de la Maza, M.J. Murtagh J.D. Spargo, and M.J. Ferraro.: Species identifies of enterococci isolated from clinical specimens , J.Clin.Microbiol, 28, 435-437 (1990)
  • Cruz, M.R., Graham, C.E., Gagliano, B.C., Lorenz, M.C., Garsin, D.A.: Enterococcus faecalis inhibits hyphal morphogenesis and virulence of Candida albicans, Infect Immun, 81(1), 189-200, (2013)
  • Hidron, A.I., Edwards, J.R., Patel, J., Horan, T.C., Sievert, D.M., Pollock, D.A., Fridkin, S.K.: NHSN annual update: antimicrobial-resistant pathogens associated with healthcare- associated infections: annual summary of data reported to the National Healthcare Safety Network at the Centers for Disease Control and Prevention, 2006-2007. Infect. Control Hosp. Epidemiol, 29, 996 –1011 (2008)
  • Crump, J.A., Collignon, P.J.: Intravascular catheter-associated infections. Eur. J. Clin. Microbiol. Infect. Dis, 19, 1–8 (2000)
  • Wisplinghoff, H., Bischoff, T., Tallent, S.M., Seifert, H., Wenzel, R.P., Edmond, M.B. : Nosocomial bloodstream infections in US hospitals: analysis of 24,179 cases from a prospective nationwide surveillance study, Clin. Infect. Dis, 39, 309–317 (2004)
  • Peciuliene, V., Reynaud, A.H., Balciuniene, I., Haapasalo, M.: Isolation of yeasts and enteric bacteria in root-filled teeth with chronic apical periodontitis. Int. Endod. J, 34, 429–434 (2001)
  • Wade, J.J., Rolando, N., Hayllar, K., Philpott-Howard, J., Casewell. M.W., Williams, R.: Bacterial and fungal infections after liver transplantation: an analysis of 284 patients. Hepatology, 21, 1328 –1336 (1995)
  • Mason, K.L., Erb Downward, J.R., Falkowski, N.R., Young, V.B., Kao, J.Y., Huffnagle, G.B.: Interplay between the gastric bacterial microbiota and Candida albicans during postantibiotic recolonization and gastritis, Infect. Immun, 80, 150 –158 (2012)
  • Mohamed, J.A., Huang, W., Nallapareddy, S.R., Teng, F., Murray, B.E.: Influence of origin of isolates, especially endocarditis isolates, and various genes on biofilm formation by Enterococcus faecalis, Infect Immun, 72, 3658–3663, (2004)
  • Mohamed, J.A., Huang, W.: Biofilm formation by Enterococci, J Med Microbiol, 56, 1581- 1588, (2007)
  • Di Rosa, R., Creti, R., Venditti, M., D’Amelio, R., Arciola, C.R., Montanaro, L., Baldassarri, L.: Relationship between biofilm formation, the enterococcal surface protein (Esp) and gelatinase in clinical isolates of Enterococcus faecalis and Enterococcus faecium. FEMS Microbiol Lett, 256, 145-150 (2006)
  • Hancock L.E., Perego M.: The Enterococcus faecalis fsr two-component system controls biofilm development through production production of gelatinase, J Bacteriol, 23, 7951- 7958 (2004)
  • Extremina, C.I., Costa, L., Aguiar, A.I., Peixe, L., Fonseca, A.P.: Optimization of processing conditions for the quantification of Enterococci biofilms using microtitre-plates, J Microbiol Meth, 84, 167-173 (2011)
  • Wang, L., Dong, M., Zheng, J., Song, Q., Yin, W., Li, J., Niu, W.: Relationship of biofilm formation and gelE gene expression in Enterococcus faecalis recovered from root canals in patients requiring endodontic retreatment, J Endod, 37, 631-636 (2011)
  • Livak, K.J., Schmittgen, T.D.: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method, Methods, 25, 402-408 (2001)
  • Adam, B., Baillie, G.S., Douglas, L.J.: Mixed species biofilms of Candida albicans and Staphylococcus epidermidis, J. Med. Microbiol, 51, 344– 349 (2002)
  • El-Azizi, M.A., Starks S.E., Khardori N.: Interactions of Candida albicans with other Candida spp. and bacteria in the biofilms, J. Appl. Microbiol, 96, 1067–1073 (2004)
  • Harriott, M.M., Noverr, M.C.: Candida albicans and Staphylococcus aureus form polymicrobial biofilms: effects on antimicrobial resistance. Antimicrob Agents Chemother, 53, 3914–3922 (2009)
  • Harriott, M.M., Noverr, M.C.: Ability of Candida albicans mutants to induce Staphylococcus aureus vancomycin resistance during polymicrobial biofilm formation, Antimicrob Agents Chemother, 54, 3746–3755 (2010)
  • Harriott, M.M., Noverr, M.C.: Importance of Candida-bacterial polymicrobial biofilms in disease, Trends Microbiol, 19, 557–563 (2011)
  • Hoffman, L.R., Deziel, E., D’Argenio, D.A., Lepine, F., Emerson, J., McNamara, S., Gibson, R.L., Ramsey, B.W., Miller, S.I.: Selection for Staphylococcus aureus small-colony variants due to growth in the presence of Pseudomonas aeruginosa, Proc Natl Acad Sci USA, 103, 19890–19895 (2006)
  • Mohamed, J.A., Huang, W., Nallapareddy, S.R., Teng, F., Murray, B.E: Influence of origin of isolates, especially endocarditis isolates, and various genes on biofilm formation by Enterococcus faecalis, Infect Immun, 72(6), 3658-3663 (2004)
  • Kristich, C.J., Li, Y.H., Cvitkovitch, D.G., Dunny, G.M. : Esp-independent biofilm formation by Enterococcus faecalis, J. Bacteriol, 186(1), 154-163 (2004)
  • Wang, L., Dong, M., Zheng, J., Song, Q., Yin, W., Li, J., & Niu, W.: Relationship of biofilm formation and gelE gene expression in Enterococcus faecalis recovered from root canals in patients requiring endodontic retreatment, J. Endod, 37(5), 631-636 (2011)
  • Liu, H., Wei, X., Ling, J., Wang, W., Huang, X.: Biofilm formation capability of Enterococcus faecalis cells in starvation phase and its susceptibility to sodium hypochlorite, J Endod, 36, 630–635 (2010)
  • Mohamed, J.A., Huang, W., Nallapareddy, S.R., Teng, F., Murray, B.E.: Influence of origin of isolates, especially endocarditis isolates, and various genes on biofilm formation by Enterococcus faecalis, Infect Immun, 72, 3658–3663 (2004)
  • Kristich, J.K., Li, Y.H., Cvitkovitch, D.G., Dunny, G.M.: Esp-independent biofilm formation by Enterococcus faecalis, J Bacteriol, 186, 154–163 (2004)
  • Seno, Y., Kariyama, R., Mitsuhata, R., Monden, K., Kumon, H.: Clinical implications of biofilm formation by Enterococcus faecalis in the urinary tract, Acta Med Okayama, 59, 79–87 (2005)
  • Rosa, D.R., Creti, R., Venditti, M., et al.: Relationship between biofilm formation, the enterococcal surface protein (Esp) and gelatinase in clinical isolates of Enterococcus faecalis and Enterococcus faecium, FEMS Microbiol Lett, 256, 145–150 (2006)
  • Duggan, J.M., Sedgley, C.M.: Biofilm formation of oral and endodontic Enterococcus faecalis, J Endod, 33, 815–818 (2007)

Determination of Gelatinase gelE Expression Levels of Enterococcus faecalis in Mono and Dual Species Biofi lms

Year 2015, Issue: 2, 162 - 173, 01.06.2015

Abstract

Biofilms are substantial problem threatening human health. Recently, it was shown that most of biofilms leading to persistent infections were polymicrobial. E. faecalis is an opportunistic pathogen frequently isolated from hospital infections such as urinary tract, wound infections etc. Differences on expression levels of some virulence factors of E.faecalis were determined between biofilm producing and nonproducing isolates. Gelatinase, a metallo protease enzyme hydrolyzing gelatin, collagen and caseine, is one of the virulence factors of E.faecalis. The role of gelatinase with biofilm formation was found in some studies using mutant strains. Our first aim is to determine the prevalence of biofilm forming ability of 11 E.faecalis urine isolates. Our second aim is to set up an in vitro dual biofilm model in a repeatable style and to find out the impact of the presence of C.albicans on the gelE gene expression of E.faecalis by using RT-qPCR. In the present study, all isolates were found as biofilm producer by both method. Moreover, our results showed increased gelE expression in dual species biofilm formed by C. albicans ATCC 90028 and E. faecalis ATCC 29212 when we compared with E. faecalis monomicrobial biofilm form. Further studies with clinical isolates will be needed to support this increased gelE expression level in the dual biofilm.

References

  • Choe, H.S., Son, S.W., Choi, H.A., Kim, H.J., Ahn, S.G., Bang, J.H., Lee, S.J., Lee, J.Y., Cho, Y.H., Lee, S.S.: Analysis of the distribution of bacteria within urinary catheter biofilms using four different molecular techniques, Am J Infect Control, 40, e249-254 (2012)
  • Kart, D., Tavernier, S., Van Acker, H., Nelis, H. J., Coenye, T.: Activity of disinfectants against multispecies biofilms formed by Staphylococcus aureus, Candida albicans and Pseudomonas aeruginosa, Biofouling, 30(3) , 377-383 (2014)
  • Yang, L., Liu, Y., Wu, H., Hİiby, N., Molin, S., Song, Z.J.: Current understanding of multi- species biofilms, IJOS, 3, 2, 74 (2011)
  • Wolcott, R., Costerton, J.W., Raoult, D., Cutler, S.J.: The polymicrobial nature of biofilm infection. Clin Microbiol Infect, 19(2), 107-112 (2013)
  • Weimer, K.E., Juneau, R.A., Murrah, K.A. et al.: Divergent mechanisms for passive pneumococcal resistance to beta-lactam antibiotics in the presence of Haemophilus influenzae, J Infect Dis, 203, 549–555 (2011)
  • Elias, S., Banin, E.: Multi-species biofilms: living with friendly neighbors, FEMS Microbiol Rev, Doi: 10.1111/j.1574-6976.2012.00325.x. [Epub ahead of print] (2012)
  • Fischbach, M.A., Sonnenburg, J.L.: Eating for two: how metabolism estab- lishes interspecies interactions in the gut, Cell Host Microbe, 10, 336–347 (2011)
  • Carlsson, J. : Bacterial metabolism in dental biofilms, Adv Dent Res, 11, 75–80 (1997)
  • Madsen, J.S., Burmolle, M., Hansen, L.H., Sorensen, S.J.: The interconnection between biofilm formation and horizontal gene transfer, FEMS Immunol Med Microbiol, 65, 183–195 (2012)
  • Arciola, C.R., Baldassarri, L., Campoccia, D., Creti, R., Prini, V., Huebner, J., Montanaro, L.: Strong biofilm production, antibiotic multi-resistance and high gelE expression in epidemic clones of Enterococcus faecalis from orthopaedic implant infections, Biomaterials, 29, 580- 586 (2008)
  • Ruoff, K.L., de la Maza, M.J. Murtagh J.D. Spargo, and M.J. Ferraro.: Species identifies of enterococci isolated from clinical specimens , J.Clin.Microbiol, 28, 435-437 (1990)
  • Cruz, M.R., Graham, C.E., Gagliano, B.C., Lorenz, M.C., Garsin, D.A.: Enterococcus faecalis inhibits hyphal morphogenesis and virulence of Candida albicans, Infect Immun, 81(1), 189-200, (2013)
  • Hidron, A.I., Edwards, J.R., Patel, J., Horan, T.C., Sievert, D.M., Pollock, D.A., Fridkin, S.K.: NHSN annual update: antimicrobial-resistant pathogens associated with healthcare- associated infections: annual summary of data reported to the National Healthcare Safety Network at the Centers for Disease Control and Prevention, 2006-2007. Infect. Control Hosp. Epidemiol, 29, 996 –1011 (2008)
  • Crump, J.A., Collignon, P.J.: Intravascular catheter-associated infections. Eur. J. Clin. Microbiol. Infect. Dis, 19, 1–8 (2000)
  • Wisplinghoff, H., Bischoff, T., Tallent, S.M., Seifert, H., Wenzel, R.P., Edmond, M.B. : Nosocomial bloodstream infections in US hospitals: analysis of 24,179 cases from a prospective nationwide surveillance study, Clin. Infect. Dis, 39, 309–317 (2004)
  • Peciuliene, V., Reynaud, A.H., Balciuniene, I., Haapasalo, M.: Isolation of yeasts and enteric bacteria in root-filled teeth with chronic apical periodontitis. Int. Endod. J, 34, 429–434 (2001)
  • Wade, J.J., Rolando, N., Hayllar, K., Philpott-Howard, J., Casewell. M.W., Williams, R.: Bacterial and fungal infections after liver transplantation: an analysis of 284 patients. Hepatology, 21, 1328 –1336 (1995)
  • Mason, K.L., Erb Downward, J.R., Falkowski, N.R., Young, V.B., Kao, J.Y., Huffnagle, G.B.: Interplay between the gastric bacterial microbiota and Candida albicans during postantibiotic recolonization and gastritis, Infect. Immun, 80, 150 –158 (2012)
  • Mohamed, J.A., Huang, W., Nallapareddy, S.R., Teng, F., Murray, B.E.: Influence of origin of isolates, especially endocarditis isolates, and various genes on biofilm formation by Enterococcus faecalis, Infect Immun, 72, 3658–3663, (2004)
  • Mohamed, J.A., Huang, W.: Biofilm formation by Enterococci, J Med Microbiol, 56, 1581- 1588, (2007)
  • Di Rosa, R., Creti, R., Venditti, M., D’Amelio, R., Arciola, C.R., Montanaro, L., Baldassarri, L.: Relationship between biofilm formation, the enterococcal surface protein (Esp) and gelatinase in clinical isolates of Enterococcus faecalis and Enterococcus faecium. FEMS Microbiol Lett, 256, 145-150 (2006)
  • Hancock L.E., Perego M.: The Enterococcus faecalis fsr two-component system controls biofilm development through production production of gelatinase, J Bacteriol, 23, 7951- 7958 (2004)
  • Extremina, C.I., Costa, L., Aguiar, A.I., Peixe, L., Fonseca, A.P.: Optimization of processing conditions for the quantification of Enterococci biofilms using microtitre-plates, J Microbiol Meth, 84, 167-173 (2011)
  • Wang, L., Dong, M., Zheng, J., Song, Q., Yin, W., Li, J., Niu, W.: Relationship of biofilm formation and gelE gene expression in Enterococcus faecalis recovered from root canals in patients requiring endodontic retreatment, J Endod, 37, 631-636 (2011)
  • Livak, K.J., Schmittgen, T.D.: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method, Methods, 25, 402-408 (2001)
  • Adam, B., Baillie, G.S., Douglas, L.J.: Mixed species biofilms of Candida albicans and Staphylococcus epidermidis, J. Med. Microbiol, 51, 344– 349 (2002)
  • El-Azizi, M.A., Starks S.E., Khardori N.: Interactions of Candida albicans with other Candida spp. and bacteria in the biofilms, J. Appl. Microbiol, 96, 1067–1073 (2004)
  • Harriott, M.M., Noverr, M.C.: Candida albicans and Staphylococcus aureus form polymicrobial biofilms: effects on antimicrobial resistance. Antimicrob Agents Chemother, 53, 3914–3922 (2009)
  • Harriott, M.M., Noverr, M.C.: Ability of Candida albicans mutants to induce Staphylococcus aureus vancomycin resistance during polymicrobial biofilm formation, Antimicrob Agents Chemother, 54, 3746–3755 (2010)
  • Harriott, M.M., Noverr, M.C.: Importance of Candida-bacterial polymicrobial biofilms in disease, Trends Microbiol, 19, 557–563 (2011)
  • Hoffman, L.R., Deziel, E., D’Argenio, D.A., Lepine, F., Emerson, J., McNamara, S., Gibson, R.L., Ramsey, B.W., Miller, S.I.: Selection for Staphylococcus aureus small-colony variants due to growth in the presence of Pseudomonas aeruginosa, Proc Natl Acad Sci USA, 103, 19890–19895 (2006)
  • Mohamed, J.A., Huang, W., Nallapareddy, S.R., Teng, F., Murray, B.E: Influence of origin of isolates, especially endocarditis isolates, and various genes on biofilm formation by Enterococcus faecalis, Infect Immun, 72(6), 3658-3663 (2004)
  • Kristich, C.J., Li, Y.H., Cvitkovitch, D.G., Dunny, G.M. : Esp-independent biofilm formation by Enterococcus faecalis, J. Bacteriol, 186(1), 154-163 (2004)
  • Wang, L., Dong, M., Zheng, J., Song, Q., Yin, W., Li, J., & Niu, W.: Relationship of biofilm formation and gelE gene expression in Enterococcus faecalis recovered from root canals in patients requiring endodontic retreatment, J. Endod, 37(5), 631-636 (2011)
  • Liu, H., Wei, X., Ling, J., Wang, W., Huang, X.: Biofilm formation capability of Enterococcus faecalis cells in starvation phase and its susceptibility to sodium hypochlorite, J Endod, 36, 630–635 (2010)
  • Mohamed, J.A., Huang, W., Nallapareddy, S.R., Teng, F., Murray, B.E.: Influence of origin of isolates, especially endocarditis isolates, and various genes on biofilm formation by Enterococcus faecalis, Infect Immun, 72, 3658–3663 (2004)
  • Kristich, J.K., Li, Y.H., Cvitkovitch, D.G., Dunny, G.M.: Esp-independent biofilm formation by Enterococcus faecalis, J Bacteriol, 186, 154–163 (2004)
  • Seno, Y., Kariyama, R., Mitsuhata, R., Monden, K., Kumon, H.: Clinical implications of biofilm formation by Enterococcus faecalis in the urinary tract, Acta Med Okayama, 59, 79–87 (2005)
  • Rosa, D.R., Creti, R., Venditti, M., et al.: Relationship between biofilm formation, the enterococcal surface protein (Esp) and gelatinase in clinical isolates of Enterococcus faecalis and Enterococcus faecium, FEMS Microbiol Lett, 256, 145–150 (2006)
  • Duggan, J.M., Sedgley, C.M.: Biofilm formation of oral and endodontic Enterococcus faecalis, J Endod, 33, 815–818 (2007)
There are 40 citations in total.

Details

Primary Language English
Journal Section Research Article
Authors

Didem Kart This is me

Publication Date June 1, 2015
Published in Issue Year 2015 Issue: 2

Cite

Vancouver Kart D. Determination of Gelatinase gelE Expression Levels of Enterococcus faecalis in Mono and Dual Species Biofi lms. HUJPHARM. 2015(2):162-73.