Biofilm Formation by Environmental Microbes Isolated from Hospitals in Karachi, Pakistan
Keywords:
Biofilm, Bacteria, Fungi, Hospital acquired infection.Abstract
The purpose of this study was to isolate and identify the microbes from hospital environmental samples and to evaluate the potential of biofilm formation by isolated microbes. For this, 125 surface swabs of different environmental samples were taken from PNS SHIFA hospital, Karachi, Pakistan. Bacteria and fungi were isolated and identified by culture plate method. Trypticase soy broth (TBS) media was used for biofilm development by microbes in plastic tubes. Developed biofilm in tubes was visualized with crystal violet staining method and then biofilm forming potential was estimated by measuring the optical density through spectrophotometer. Total 202 microbes including 126(62.38%) bacteria and 76(37.62%) fungi were isolated and identified. Among environmental samples, hospital ward curtains and medical trays were highly contaminated with bacteria and fungi (with 26% each of total assemblage respectively). Staphylococcus aureus was in highest abundance followed by Candida albicans with 28.7% and 15.8% of total assemblage of isolation respectively. Moreover; Staphylococcus aureus followed by Candida albicans also found to have highest potential to form biofilm with 30.25% and 23.52% of total assemblage of biofilm formation respectively which clearly indicates that Staphylococcus aureus and Candida albicans may recognized as major agents of hospital acquired infection and can relate with enhanced potential of biofilm formation. Highest abundance and biofilm formation potential of bacteria and fungi in combination can also underline a direct extensive and striking interaction between prokaryotic and eukaryotic cells in biofilm.
References
[1] J.W. Costerton., P.S. Stewart. and E.P. Greenberg. “Bacterial biofilms: a common cause of persistent Infections”. Science,Vol. 284, pp. 1318-1322, 1999.
[2] L. Hall-Stoodley. and P. Stoodley. “Biofilm formation and dispersal and the transmission of human pathogens”. Trends Microbiol, vol. 13, pp. 7-10, 2005.
[3] D. Lindsay. and A.von Holy. “Bacterial biofilms within the clinical setting: what healthcare professionals should know”. J Hosp Infec, vol. 64, pp. 313-325, 2006.
[4] A. Bridier, R. Briandet, V. Thomas. and F. Dubois-Brissonnet. “Resistance of bacterial biofilms to disinfectants: a review”. Biofouling, vol. 27, pp. 1017–1032, 2011.
[5] T.F. Mah. and G.A. O’Toole. “Mechanisms of biofilm resistance to antimicrobial agents”. Trends Microbiol, vol. 9, pp. 34-39, 2001.
[6] H. Lee, Y. Koh, J. Kim, J.C. Lee, Y. C.Lee, S.Y. Seol, D.T. Cho. and J. Kim. “Capacity of multidrug- resistant clinical isolates of Acinetobacter baumannii to form biofilm and adhere to epithelial cell surfaces”. Clin Microbiol Infect, vol. 14, pp. 49-54, 2008.
[7] N. Hoiby, T. Bjarnsholt, M. Givskov, S. Molin, O. Ciofu. “Antibiotic resistance of bacterial biofilms”. Int J Antimicrob Agents, vol. 35, pp. 322-332, 2010.
[8] K. Smith, and I.S. Hunter. “Efficacy of common hospital biocides with biofilms of multi-drug resistant clinical isolates”. J Med Microbiol, vol. 57, pp. 966-973, 2008.
[9] A. Kramer, I. Schwebke, G. Kampf. “How long do nosocomial pathogens persist on inanimate surfaces? A systematic review”. BMC Infect Dis, vol. 6, pp. 130, 2006.
[10] J.A. Otter, S. Yezli, and G.L. French. “The role played by contaminated surfaces in the transmission of nosocomial pathogens”. Infect Control Hosp Epidemiol, vol.32, pp. 687-699, 2011.
[11] S. Martí, J. Rodríguez-Baño, M. Catel-Ferreira, T. Jouenne, J. Vila, H. Seifert, E. Dé. “Biofilm formation at the solid-liquid and air-liquid interfaces by Acinetobacter species”. BMC Res. Notes, vol. 4, pp. 5, 2011.
[12] D.J. Stickler. “Susceptibility of antibiotic-resistant Gram positive bacteria to biocides: a perspective from the study of catheter biofilms”. Symp Ser Soc Appl Microbiol, vol. 31, pp. 163S–170S, 2002.
[13] J.L. Vincent. “Nosocomial infections in adult intensive-care units”. Lancet, vol. 361, pp. 2068–2077, 2003.
[14] J. Treter, A.J. Macedo. “Catheters. A suitable surface for biofilm formation”. In: Méndez-Vilas A. editor. Handbook: science against microbial pathogens: communicating current research and technological advances. Spain: Formatex Research Center; 2011, pp. 835-842.
[15] C.R Kokare, S. Chakraborty, A.N. Khopade, K.R. Mahadik. “Biofilm: importance and applications”. Indian J Biotechnol, vol. 8, pp. 159-68, 2009.
[16] N.F. Al-saad, M.S. Al-saeed and I. A. bnyab. “Biofilm formation by bacteria isolates from burn infected patients”. Med J Babylon vol. 9(3), pp. 517-525, 2012.
[17] C. Hedayati Potera. “Forging a link between biofilms and disease”. Science, vol. 283, pp. 1837–1838, 1999.
[18] J.A. Lichter, K.J. Van Vliet and M.F. Rubner. “Design of antibacterial surfaces and interfaces: polyelectrolyte multilayers as a multifunctional platform”. Macromolecules, vol. 42, pp. 8573-8586, 2009.
[19] K. Vickery, A. Deva, A. Jacombs, J. Allan, P. Valente and I.B. Gosbell. “Presence of biofilm containing viable multiresistant organisms despite terminal cleaning on clinical surfaces in an intensive care unit”. J Hosp Infect, vol. 80, pp. 52-55, 2012.
[20] S. Eftekhar F. Hosseini S.M. “Biofilm formation by bacteria isolated from intravenous catheters”. J med bacterial, vol. 3(3), pp. 26-31, 2014.
[21] J.S. Peng, W.C. Tsai and C.C. Chou. “Inactivation and removal of Bacillus cereus by sanitizer and detergent”. Int J Food Microbiol, vol. 77, pp. 11-18, 2002.
[22] F.A. Manian, S. Griesenauer and D. Senkel. “Isolation of Acinetobacter baumannii complex and methicillin-resistant Staphylococcus aureus from hospital rooms following terminal cleaning and disinfection: can we do better?”. Infect Control Hosp Epidemiol, vol. 32, pp. 667-672, 2011.
[23] S. Gander. “Bacterial biofilms: resistance to antimicrobial agents”. J Antimicrob Chemother, vol. 37, pp. 1047–1050, 1996.
[24] P. Gilbert, D.G. Allison and A.J. McBain. “Biofilms in vitro and in vivo: do singular mechanisms simply cross-resistance?” J Appl Microbiol, vol. 92, pp. 98S–110S, 2002.
[25] G.A. O’Toole. “A resistance switch”. Nature, vol. 416, pp. 695–696, 2002.
[26] H.F. Jenkinson, and L.J. Douglas. “Interactions between Candida species and bacteria in mixed infections”. In: Brogden KA, Guthmiller JM, editors. Handbook of: Polymicrobial Diseases. ASM Press, 2002, p. 2357–373.
[27] R.M. Donlan. “Biofilms and device associated infections”. Emerg Infect Dis, vol. 7(2), pp. 277-281, 2001.
[28] C.A. Fux, J.W. Costerton, P.S. Stewart and P. Stoodley. “Survival strategies of infectious biofilms”. Trends Microbiol, vol. 13, pp. 34-40, 2005.
[29] R.A. Calderone. “Introduction and historical perspectives”. In: Calderone RA editor. Handbook: Candida and Candidiasis, ASM Press, 2002, pp. 3–13.
[30] C.G. Adair. “Implications of endotracheal tube biofilm ventilator-associated pneumonia”. Intensive Care Med, vol. 25, pp. 1072–1076, 1999.
[31] S.P. Hawser, and L.J. Douglas. ‘Biofilm formation by Candida species on the surface of catheter materials in vitro’. Infect Immun, vol. 62, pp. 915–921, 1994.
[32] H. Nikawa. “The role of saliva and serum in Candida albicans biofilm formation on denture acrylic surfaces”. Microb Ecol Health Dis, vol. 9, pp. 35–48, 1996.
[33] G. Ramage. “Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms”. Antimicrob Agents Chemother,vol. 45, pp. 2475–2479, 2001.
[34] J. Biedlingmaier, R. Samaranayake, and P. Whelan. “Resistance to biofilm formation on otologic implant materials”. Otolaryngol Head Neck Surg, vol.118, pp. 444–451, 1998.
[35] G.D. Christensen, W.A. Simpson, and J.J. Younger. “Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices”. J Clin Microbiol, vol. 22 (6), pp. 996–1006, 1985.
[36] X. Li, Z. Yan, and J. Xu. “Quantitative variation of biofilms among strains in natural populations of Candida albicans”. Microbiology, vol. 149(2), pp. 353–362, 2003.
[37] S. Stepanovi´c, D. Vukovi´c, I. Daki´c, B. Savi´c, and M. ?Svabi´c-Vlahovi´c. “A modified microtiter-plate test for quantification of staphylococcal biofilm formation”. J Microbiol Methods, vol. 40(2), pp. 175–179, 2000.
[38] L. Cr´emet, S. Corvec, and E. Batard. “Comparison of three methods to study biofilm formation by clinical strains of Escherichia coli”. Diag Microbiol Infect Dis. Vol.75(3), pp. 252–255, 2013.
[39] J.G. Holt, N.R. Krieg, P.H.A. Sneath, and J.T. Staley. “Bergey’s Manual of Determinative Bacteriology”. 9th ed. Williams and Wilkins Company, Baltimore, 1994, 255-273.
[40] Philos. “Practical Mycology: Manual for identification of fungi”. A.W. BrØggers Boktrykker, A/S. 1961.
[41] J.A. Otter, J. Chewins, S. Yezli, and J.D. Edgeworth. In Press: “Surface-attached cells, biofilms and biocide susceptibility: implications for hospital cleaning and disinfection”. J. Hosp Infec, pp.1-12, 2014.
[42] Stickler. “Bacterial biofilms and the encrustations of urethral catheter”. Biofouling, vol. 9(4), pp. 293-305, 1996.
[43] R. Vansanthi. D. Karthikeyan. and M. Jeya. ‘Study of biofilm production and antimicrobial resistance pattern of the bacterial isolates from invasive devices”. Int J Res Health Sci, vol. 2(1), pp. 274-81, 2014.
[44] E. Curran, J.E. Coia, and H. Gilmour. “Multi-centre research surveillance project to reduce infections/phlebitis associated with peripheral vascular catheters”. J Hosp Infect, vol. 46 (3), pp. 194-202, 2000.
[2] L. Hall-Stoodley. and P. Stoodley. “Biofilm formation and dispersal and the transmission of human pathogens”. Trends Microbiol, vol. 13, pp. 7-10, 2005.
[3] D. Lindsay. and A.von Holy. “Bacterial biofilms within the clinical setting: what healthcare professionals should know”. J Hosp Infec, vol. 64, pp. 313-325, 2006.
[4] A. Bridier, R. Briandet, V. Thomas. and F. Dubois-Brissonnet. “Resistance of bacterial biofilms to disinfectants: a review”. Biofouling, vol. 27, pp. 1017–1032, 2011.
[5] T.F. Mah. and G.A. O’Toole. “Mechanisms of biofilm resistance to antimicrobial agents”. Trends Microbiol, vol. 9, pp. 34-39, 2001.
[6] H. Lee, Y. Koh, J. Kim, J.C. Lee, Y. C.Lee, S.Y. Seol, D.T. Cho. and J. Kim. “Capacity of multidrug- resistant clinical isolates of Acinetobacter baumannii to form biofilm and adhere to epithelial cell surfaces”. Clin Microbiol Infect, vol. 14, pp. 49-54, 2008.
[7] N. Hoiby, T. Bjarnsholt, M. Givskov, S. Molin, O. Ciofu. “Antibiotic resistance of bacterial biofilms”. Int J Antimicrob Agents, vol. 35, pp. 322-332, 2010.
[8] K. Smith, and I.S. Hunter. “Efficacy of common hospital biocides with biofilms of multi-drug resistant clinical isolates”. J Med Microbiol, vol. 57, pp. 966-973, 2008.
[9] A. Kramer, I. Schwebke, G. Kampf. “How long do nosocomial pathogens persist on inanimate surfaces? A systematic review”. BMC Infect Dis, vol. 6, pp. 130, 2006.
[10] J.A. Otter, S. Yezli, and G.L. French. “The role played by contaminated surfaces in the transmission of nosocomial pathogens”. Infect Control Hosp Epidemiol, vol.32, pp. 687-699, 2011.
[11] S. Martí, J. Rodríguez-Baño, M. Catel-Ferreira, T. Jouenne, J. Vila, H. Seifert, E. Dé. “Biofilm formation at the solid-liquid and air-liquid interfaces by Acinetobacter species”. BMC Res. Notes, vol. 4, pp. 5, 2011.
[12] D.J. Stickler. “Susceptibility of antibiotic-resistant Gram positive bacteria to biocides: a perspective from the study of catheter biofilms”. Symp Ser Soc Appl Microbiol, vol. 31, pp. 163S–170S, 2002.
[13] J.L. Vincent. “Nosocomial infections in adult intensive-care units”. Lancet, vol. 361, pp. 2068–2077, 2003.
[14] J. Treter, A.J. Macedo. “Catheters. A suitable surface for biofilm formation”. In: Méndez-Vilas A. editor. Handbook: science against microbial pathogens: communicating current research and technological advances. Spain: Formatex Research Center; 2011, pp. 835-842.
[15] C.R Kokare, S. Chakraborty, A.N. Khopade, K.R. Mahadik. “Biofilm: importance and applications”. Indian J Biotechnol, vol. 8, pp. 159-68, 2009.
[16] N.F. Al-saad, M.S. Al-saeed and I. A. bnyab. “Biofilm formation by bacteria isolates from burn infected patients”. Med J Babylon vol. 9(3), pp. 517-525, 2012.
[17] C. Hedayati Potera. “Forging a link between biofilms and disease”. Science, vol. 283, pp. 1837–1838, 1999.
[18] J.A. Lichter, K.J. Van Vliet and M.F. Rubner. “Design of antibacterial surfaces and interfaces: polyelectrolyte multilayers as a multifunctional platform”. Macromolecules, vol. 42, pp. 8573-8586, 2009.
[19] K. Vickery, A. Deva, A. Jacombs, J. Allan, P. Valente and I.B. Gosbell. “Presence of biofilm containing viable multiresistant organisms despite terminal cleaning on clinical surfaces in an intensive care unit”. J Hosp Infect, vol. 80, pp. 52-55, 2012.
[20] S. Eftekhar F. Hosseini S.M. “Biofilm formation by bacteria isolated from intravenous catheters”. J med bacterial, vol. 3(3), pp. 26-31, 2014.
[21] J.S. Peng, W.C. Tsai and C.C. Chou. “Inactivation and removal of Bacillus cereus by sanitizer and detergent”. Int J Food Microbiol, vol. 77, pp. 11-18, 2002.
[22] F.A. Manian, S. Griesenauer and D. Senkel. “Isolation of Acinetobacter baumannii complex and methicillin-resistant Staphylococcus aureus from hospital rooms following terminal cleaning and disinfection: can we do better?”. Infect Control Hosp Epidemiol, vol. 32, pp. 667-672, 2011.
[23] S. Gander. “Bacterial biofilms: resistance to antimicrobial agents”. J Antimicrob Chemother, vol. 37, pp. 1047–1050, 1996.
[24] P. Gilbert, D.G. Allison and A.J. McBain. “Biofilms in vitro and in vivo: do singular mechanisms simply cross-resistance?” J Appl Microbiol, vol. 92, pp. 98S–110S, 2002.
[25] G.A. O’Toole. “A resistance switch”. Nature, vol. 416, pp. 695–696, 2002.
[26] H.F. Jenkinson, and L.J. Douglas. “Interactions between Candida species and bacteria in mixed infections”. In: Brogden KA, Guthmiller JM, editors. Handbook of: Polymicrobial Diseases. ASM Press, 2002, p. 2357–373.
[27] R.M. Donlan. “Biofilms and device associated infections”. Emerg Infect Dis, vol. 7(2), pp. 277-281, 2001.
[28] C.A. Fux, J.W. Costerton, P.S. Stewart and P. Stoodley. “Survival strategies of infectious biofilms”. Trends Microbiol, vol. 13, pp. 34-40, 2005.
[29] R.A. Calderone. “Introduction and historical perspectives”. In: Calderone RA editor. Handbook: Candida and Candidiasis, ASM Press, 2002, pp. 3–13.
[30] C.G. Adair. “Implications of endotracheal tube biofilm ventilator-associated pneumonia”. Intensive Care Med, vol. 25, pp. 1072–1076, 1999.
[31] S.P. Hawser, and L.J. Douglas. ‘Biofilm formation by Candida species on the surface of catheter materials in vitro’. Infect Immun, vol. 62, pp. 915–921, 1994.
[32] H. Nikawa. “The role of saliva and serum in Candida albicans biofilm formation on denture acrylic surfaces”. Microb Ecol Health Dis, vol. 9, pp. 35–48, 1996.
[33] G. Ramage. “Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms”. Antimicrob Agents Chemother,vol. 45, pp. 2475–2479, 2001.
[34] J. Biedlingmaier, R. Samaranayake, and P. Whelan. “Resistance to biofilm formation on otologic implant materials”. Otolaryngol Head Neck Surg, vol.118, pp. 444–451, 1998.
[35] G.D. Christensen, W.A. Simpson, and J.J. Younger. “Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices”. J Clin Microbiol, vol. 22 (6), pp. 996–1006, 1985.
[36] X. Li, Z. Yan, and J. Xu. “Quantitative variation of biofilms among strains in natural populations of Candida albicans”. Microbiology, vol. 149(2), pp. 353–362, 2003.
[37] S. Stepanovi´c, D. Vukovi´c, I. Daki´c, B. Savi´c, and M. ?Svabi´c-Vlahovi´c. “A modified microtiter-plate test for quantification of staphylococcal biofilm formation”. J Microbiol Methods, vol. 40(2), pp. 175–179, 2000.
[38] L. Cr´emet, S. Corvec, and E. Batard. “Comparison of three methods to study biofilm formation by clinical strains of Escherichia coli”. Diag Microbiol Infect Dis. Vol.75(3), pp. 252–255, 2013.
[39] J.G. Holt, N.R. Krieg, P.H.A. Sneath, and J.T. Staley. “Bergey’s Manual of Determinative Bacteriology”. 9th ed. Williams and Wilkins Company, Baltimore, 1994, 255-273.
[40] Philos. “Practical Mycology: Manual for identification of fungi”. A.W. BrØggers Boktrykker, A/S. 1961.
[41] J.A. Otter, J. Chewins, S. Yezli, and J.D. Edgeworth. In Press: “Surface-attached cells, biofilms and biocide susceptibility: implications for hospital cleaning and disinfection”. J. Hosp Infec, pp.1-12, 2014.
[42] Stickler. “Bacterial biofilms and the encrustations of urethral catheter”. Biofouling, vol. 9(4), pp. 293-305, 1996.
[43] R. Vansanthi. D. Karthikeyan. and M. Jeya. ‘Study of biofilm production and antimicrobial resistance pattern of the bacterial isolates from invasive devices”. Int J Res Health Sci, vol. 2(1), pp. 274-81, 2014.
[44] E. Curran, J.E. Coia, and H. Gilmour. “Multi-centre research surveillance project to reduce infections/phlebitis associated with peripheral vascular catheters”. J Hosp Infect, vol. 46 (3), pp. 194-202, 2000.
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2016-01-15
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Shaheen, A., & Baqai, R. (2016). Biofilm Formation by Environmental Microbes Isolated from Hospitals in Karachi, Pakistan. American Scientific Research Journal for Engineering, Technology, and Sciences, 15(1), 240–251. Retrieved from https://asrjetsjournal.org/index.php/American_Scientific_Journal/article/view/1238
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