Supplementary Files
Keywords
Congo red;
Multidrug resistance;
Virulence.
How to Cite
Abstract
Background: Biofilm is defined as an assembly of microorganisms which enclosed in a self-produced extracellular matrix principally of polysaccharide material and found in association with indwelling medical devices. This study was designed to determine the biofilm forming ability of isolates from devices associated infection.
Objectives: To compare and evaluate biofilm production with the virulence markers like multidrug resistance and slime production.
Methods: An analytical observational study was conducted in Manipal Teaching Hospital from 2020 June-2021 May after ethical clearance. A total of 106 clinical isolates were obtained from patients with indwelling medical devices. All bacteria were identified by conventional techniques. Antimicrobial sensitivity testing was performed on Mueller-Hinton agar plates with commercially available antibiotic discs using Kirby Bauer disc diffusion techniques and interpreted as per the guidelines of Clinical and Laboratory Standards Institute (CLSI). Biofilm and slime production were detected by two methods: Tissue culture plate method and Congo Red Agar method.
Results: Out of 106 total isolates, 79 (74.5%) isolates were detected in endotracheal tubes (ETTs). Besides, it was observed that 54 (68.3%) of the 79 ETT isolates were biofilm producers. Amongst the isolates, 90.4% (19/21) were Klebsiella species, 64.1% (25/39) Acinetobacter spp., 47.6% (10/21) Pseudomonas spp., and 54.5% (6/11) Staphylococcus aureus were biofilm producers.
Conclusion: Biofilm mediated persistence of infection in the nosocomial setting through indwelling devices. Significantly, higher number of the biofilm producers as well as slime producers were multidrug resistant (p-value <0.05).
References
Danhorn T, Fuqua C. Biofilm formation by plant-associated bacteria. Annu Rev Microbiol. 2007;61:401-22. [PubMed | Full Text | DOI]
Assefa M, Amare A. Biofilm-associated multi-drug resistance in hospital-acquired infections: A review. Infect Drug Resist. 2022;15:5061-8. [PubMed | Full Text | DOI]
Rajkumari S, Jha B, Adhikaree N. Bacteriological profile and antibiogram of endotracheal aspirates in patients admitted in neurosurgical intensive care unit at a tertiary care hospital. J Coll Med Sci Nepal. 2020;16(3):152-6. [Full Text | DOI]
Khatun MN, Shamsuzzaman SM, Fardows J, Siddique AB, Joly SN. Identification of bacterial isolates from endotracheal aspirate of patients in intensive care unit and their antimicrobial susceptibility pattern. J Enam Med Col. 2018;8(2):67-73. [Full Text]
Mansilla CA, Alarcon JM, Ahufinger IG, Ramirez MG. Microbiological diagnosis of catheter-related infections. Enferm Infecc Microbiol Clin. 2019;37(10):668-72. [PubMed | Full Text | DOI]
Collee JG, Marmion BP, Fraser AG, Simmons A, editors. Mackie and McCartney’s Practical Medical Microbiology. 14th ed. New York: Churchill Livingstone; 1996, 978. [Full Text]
Forbes BA, Sham DF, Weissfeld AS. Bailey and Scott’s Diagnostic Microbiology, 10th ed. New York: Mosby; 1998.167-87. [Full Text]
Bauer KBR, Kirby WMM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disc method. Am J Clin Pathol. 1966;45:493-6. [PubMed | Full Text | DOI]
Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG, et al. Multidrug-resistant, extensively drug-resistant, and pandrug-resistant bacteria: An international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect. 2012;18(3):268-81. [PubMed | Full Text | DOI]
Christensen GD, Simpson WA, Younger JA et al. Adherence of coagulase-negative staphylococci to plastic tissue cultures: A quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol. 1985;22:996-1006. [PubMed | Full Text | DOI]
Mathur T, Singhal S, Khan S, Upadhyay DJ, Fatma T, Rattan A. Detection of biofilm formation among the clinical isolates of staphylococci: An evaluation of three different screening methods. Indian J Med Microbiol. 2006;24(1):25-9. [PubMed | Full Text | DOI]
Stepanovic S, Vukovi D, Hola V, Di Bonaventura G, Djukic S, Cirkovi I, et al. Quantification of biofilm in microtiter plates: Overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS. 2007;115(8):891-9. [PubMed | Full Text | DOI]
Freeman J, Falkiner FR, Keane CT. New method for detecting slime production by coagulase negative staphylococci. J Clin Pathol. 1989;42:872-4. [PubMed | Full Text | DOI]
Reid G. Biofilms in infectious disease and on medical devices. Int J Antimicrob Agents. 1999;11(3-4):223-6. [PubMed | Full Text | DOI]
Shrestha LB, Baral R, Khanal B. Comparative study of antimicrobial resistance and biofilm formation among gram-positive uropathogens isolated from community-acquired urinary tract infections and catheter-associated urinary tract infections. Infect Drug Resist. 2019;12:957-63. [PubMed | Full Text | DOI]
Zmantar T, Chaieb K, Makni H, Miladi H, Abdulla FB, Mahdouni K, et al. Detection by PCR of adhesins gene and slime production in clinical staphylococcus aureus. J Basic Microbiol. 2008;48(4):308-14. [PubMed | Full Text | DOI]
Nayak N, Nag TC, Satpathy G, Ray SB. Ultrastructural analysis of slime positive and slime negative staphylococcus epidermidis isolates in infectious keratitis. Indian J Med Res. 2007;125(6):767-71. [PubMed | Full Text]
Sannathimmappa MB, Nambiar V, Aravindakshan R, Al Kasaby NM. Profile and antibiotic resistance pattern of bacteria isolated from endotracheal secretions of mechanically ventilated patients at a tertiary care hospital. J Educ Health Promot. 2021;10(1):195. [PubMed | Full Text | DOI]
Prasad S, Nayak N, Satpathy G, Nag HL, Venkatesh P, Ramakrishnan S, et al. Molecular and phenotypic characterisation of staphylococcus epidermidis in implant related infections. Indian J Med Res. 2012;136(3);483-90. [PubMed | Full Text]
Gil-Perotin S , Ramirez P, Marti V, Sahuquillo JM , Gonzalez E, Calleja I, et al. Implications of endotracheal tube biofilm in ventilator-associated pneumonia response: A state of concept. Crit Care. 2012;16(3):R93. [PubMed | Full Text | DOI]
Feldman C, Kassel M, Cantrell J, Kaka S, Morar R, Goolam Mahomed A, et al. The presence and sequence of endotracheal tube colonisation in patients undergoing mechanical ventilation. Eur Respir J. 1999;13(3):546-51. [PubMed | Full Text | DOI]
Sirvent JM, Torres A, Vidaur L, Armengol J, de Batlle J, Bonet A. Tracheal colonisation within 24 h of intubation in patients with head trauma: Risk factor for developing early-onset ventilator-associated pneumonia. Intensive Care Med. 2000;26(9):1369-72. [PubMed | Full Text | DOI]
Bonten MJM, Kollef MH, Hall JB. Risk factors for ventilator-associated pneumonia: From epidemiology to patient management. Clin Infect Dis. 2004;38(8):1141-9. [PubMed | Full Text | DOI]
Costerton JW, Stewart PS, Greenberg EP: Bacterial biofilms: A common cause of persistent infections. Science 1999;284:1318-22. [PubMed | Full Text | DOI]
Bagge N, Ciofu O, Skovgaard LT, Høiby N. Rapid development in vitro and in vivo of resistance to ceftazidime in biofilm-growing Pseudomonas aeruginosa due to chromosomal beta-lactamase. APMIS. 2000;108:589-600. [PubMed | Full Text | DOI]
Gordon CA, Hodges NA, Marriott C. Antibiotic interaction and diffusion through alginate and exopolysaccharide of cystic fibrosisderived Pseudomonas aeruginosa. J Antimicrob Chemother. 1988;22:667-74. [PubMed | Full Text | DOI]
Sharma D, Misba L, Khan AU. Antibiotics versus biofilm: An emerging battle ground in microbial communities. Antimicrob Resist Infect Control. 2019;8:76. [PubMed | Full Text | DOI]