IDENTIFICATION AND DETECTION OF BIOFILM PRODUCING STAPHYLOCOCCUS AUREUS AND ITS ANTIBIOGRAM ACTIVITIES

SAPANA SHARMA; UPASHANA BHANDARI; YOGESH OLI; GANESH BHANDARI; SUNITA BISTA; GANGA GC; BASUDHA SHRESTHA; NETRA LAL BHANDARI

doi:10.22159/ajpcr.2021.v14i4.40728

Authors

SAPANA SHARMA Department of Microbiology, Tri-Chandra Multiple Campus, Tribhuvan University, Kathmandu, Nepal.
UPASHANA BHANDARI Department of Chemistry, Tri-Chandra Multiple Campus, Tribhuvan University, Kathmandu, Nepal.
YOGESH OLI Department of Microbiology, Tri-Chandra Multiple Campus, Tribhuvan University, Kathmandu, Nepal.
GANESH BHANDARI Department of Chemistry, Tri-Chandra Multiple Campus, Tribhuvan University, Kathmandu, Nepal.
SUNITA BISTA Department of Chemistry, Tri-Chandra Multiple Campus, Tribhuvan University, Kathmandu, Nepal.
GANGA GC Department of Microbiology, Tri-Chandra Multiple Campus, Tribhuvan University, Kathmandu, Nepal.
BASUDHA SHRESTHA Department of Microbiology, Kathmandu Model Hospital, Kathmandu, Nepal.
NETRA LAL BHANDARI Department of Chemistry, Tri-Chandra Multiple Campus, Tribhuvan University, Kathmandu, Nepal.

DOI:

https://doi.org/10.22159/ajpcr.2021.v14i4.40728

Keywords:

Methicillin-resistant Staphylococcus aureus, Methicillin-sensitive Staphylococcus aureus, Biofilm, Kirby-Bauer disk diffusion, Tissue culture plate, Minimum inhibitory concentration

Abstract

Objectives: The main aim of this work is to determine the antibiogram profile of biofilm-producing Staphylococcus aureus from various clinical specimens of the patients.

Methods: Various bacterial cultures of non-repeated clinical specimens from a total of 3388 patients were determined using standard microbiological and biochemical methods.

Results: Out of 3388 only 604 (17.02%) displayed growth positive. A total of 65 (51.58%) S. aureus isolates were recovered, 25 (38.46%) were identified as methicillin-resistant S. aureus (MRSA) by Cefoxitin (30 μg) disk diffusion technique, of which majority were from pus/wound swab 22 (37.29%). The antibiogram of the isolates was analyzed by Kirby-Bauer disk diffusion technique analyzing Linezolid to be the most effective drug with susceptibility of 100% to both MRSA and methicillin-sensitive S. aureus, followed by vancomycin, tigecycline, and tetracycline. In vitro biofilm production by tissue culture plate (TCP) and Congo red agar method detected 52 (80%) and 25 (38.46%) as biofilm producers, respectively. TCP identified 2 (3.07%), 7 (10.76%), and 44 (67.69%) as strongly, moderately, and weakly adherent. About 30.7% of MRSA obtained were positive biofilm producers. The minimum inhibitory concentration value of Oxacillin for S. aureus by agar dilution method ranged from 0.025 μg/mL to 128 μg/mL.

Conclusion: This study shows that biofilm production was more in methicillin-resistant strains and displayed a high degree of resistance to almost all groups of antibiotics.

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References

Stapleton PD, Taylor PW. Methicillin resistance in Staphylococcus aureus: Mechanism and modulation. Sci Prog 2002;85:57-72.

Pozzi C, Waters EM, Rudkin JK, Lohan AM, Tong P, Loftus BJ, et al. Methicillin resistance alters the biofilm phenotype and attenuates virulences in S. aureus device-associated infections. PLoS Pathog 2012;8:e1002626.

Thammanvongsa V, Kim HK, Missiakleas D, Scheewmd O. Staphylococcal manipulation of host immune responses. Nature Rev Microbiol 2015;13:529-43.

Mathur T, Singhal S, Khan S, Upadhay 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;24:25-9.

Ziebhur W, Lobner I, Krimmer V, Hacker J. Methods to detect and analyze phenotypic variation in biofilm-forming staphylococci. Method Enzymol 2001;336:195-233.

Houston P, Rowe SE, Pozzi R, Water EM, Gara JP. Essential role for the major autolysin in the fibronectin-binding protein-mediated Staphylococcus aureus biofilm phenotype. Infect Immun 2011;79:1153-65.

Altieri KT, Sanita PV, Machado AL, Giampaolo ET, Pavarina AC, Jorge JH, et al. Eradication of mature methicillin-resistant Staphylococcus aureus (MRSA) biofilm from acrylic surfaces. Braz Dent J 2013;24:487-91.

Hadler JL, Petit S, Mandour M, Cartter ML. Trend in invasive infection with methicillin-resistant Staphylococcus aureus, Connecticut, USA, 2001-2010. Emerg Infect Dis 2012;18:917-24.

Gould IM. The clinical significance of methicillin-resistant Staphylococcus aureus. J Hosp Infect 2005;61:272-82.

Ansari S, Nepal HP, Gautam R, Rayamajhi N, Shrestha S, Upadhyay G, et al. Threat of drug-resistant Staphylococcus aureus to health in Nepal. Biomed Cent Infect Dis 2014;14:157.

Wolcott RD, Ehrlich GD. Biofilm and chronic infections. J Am Med Assoc 2008;299:2682-4.

Enright MC. The evolution of a resistant pathogen-the case of MRSA. Curr Opin Pharmacol 2003;3:474-9.

Hidron Al, Kourbatova EV, Halvosa JS, Terrell BJ, McDougal LK, Tenover FC, et al. Risk factors for colonization with methicillin-resistant Staphylococcus aureus (MRSA) in patients admitted to Urban hospital: Emergence of community-associated MRSA nasal carriage. Clin Infect Dis 2005;41:159-66.

Collee JF, Fraser AG, Marmion BO, Simmons A. Staphylococcus: Cluster forming gram positive cocci. In: Mackie and McCartney Practical Medical Microbiology. 14th ed. United States: Churchill Livingstone; 2006.

Cheesebrough M. District Laboratory Practice in Tropical Countries Part 2. Cambridge, England: Cambridge University Press; 2006.

Clinical and Laboratory Standards Institute. CLSI Performance Standards for Antimicrobial Susceptibility Testing, Twenty-fourth Supplement, CLSI Document M100-S24. Wayne, PA: Clinical and Laboratory Standards Institute; 2014.

Moghadam SO, Pourmad MR, Aminharati F. Biofilm formation and antimicrobial resistance in methicillin-resistant Staphylococcus aureus isolated from burn patients, Iran. J Infect Dev Countries 2014;8:1511-7.

Kwasny SM, Opperman JT. Static biofilm cultures of gram-positive pathogens grown in microtiter format used for anti-biofilm drug discovery. Curr Protoc Pharmacol 2010;50:13A.8.

Kshetry AO, Pant ND, Khatri S, Shrestha KL, Upadhaya SK, Poudel A, et al. Minimum inhibitory concentration of vancomycin to methicillin-resistant Staphylococcus aureus isolated from different clinical samples at a tertiary care hospital in Nepal. Antimicrob Resist Infect Control 2016;5:27.

Iregbu KC, Uwaezuoke NS, Nwajiobi-Princewill IP, Eze SO, Medugu N, Shettima S, et al. A profile of wound infection in national hospital Abuja. Afr J Clin Exp Microbiol 2013;14:160-3.

Garcia-Granja PE, Lόpez J, Vilacosta I, Ortiz-Bautista C, Sevilla T, Olmos C, et al. Polymicrobial infective endocarditis: Clinical features and prognosis. Medicine 2015;94:1-6.

Pandey S, Raza S, Bhatta CP. Prevalence and antibiotic sensitivity pattern of methicillin-resistant Staphylococcus aureus in Kathmandu medical college-Teaching hospital. J Inst Med 2012;34:13-7.

Shahina Z, Chowdhury AH, Arifuzzaman MD. Prevalence of antimicrobial sensitivity and resistant pattern of a gram-positive cluster forming cocci in clinical samples. IOSR J Dent Med Sci 2014;13:53-7.

Bhandari G, Pokharel B, Oli Y, Katuwal A, Bhandari NL. Screening of methicillin-resistant Staphylococcus aureus (MRSA) from wounds in pediatric patients visiting tertiary care in hospital. Nepal J Biotechnol 2019;7:82-9.

Belbase A, Pant ND, Nepal K, Nepal B, Baidhya R, Lekhak B. Antibiotic resistance and biofilm production among the strains of Staphylococcus aureus isolated from pus/wound swab samples in a tertiary care hospital in Nepal. Ann Clin Microbiol Antimicrob 2017;16:2-5.

Verma AK, Kapoor AK, Bhargava A. Antimicrobial susceptibility pattern of bacterial isolates from surgical wound infection in tertiary care hospital in Allahabad, India. Int J Med Update 2012;7:27-34.

Adhikari R, Pant ND, Neupane M, Bhattarai R, Bhatta S, Chaudhary R, et al. Detection of methicillin-resistant Staphylococcus aureus and determination of the minimum inhibitory concentration of vancomycin for Staphylococcus aureus isolated from pus/wound samples of the patients attending a tertiary care hospital in Kathmandu Nepal. Can J Infect Dis Med Microbiol 2017:1-6.

Tiwari HK, Das AK, Sapkota D, Sivarjan K, Pahwa VK. Methicillin-resistant Staphylococcus aureus prevalence and antibiogram in tertiary care hospital in Western Nepal. J Infect Dev Ctries 2009;3:681-4.

Goyal A, Diwakar MK, Bhosshan S, Goyal S, Agrawal A. Prevalence and antimicrobial susceptibility pattern of methicillin-resistant Staphylococcus aureus (MRSA) isolates at a tertiary care hospital in Agra, North India-a systemic annual review. IOSR J Dent Med Sci 2013;11:80-4.

Ghellai L, Hassaine H, Klouche N, Khadir A, Aissaoui N, Nas F, et al. Detection of biofilm formation of a collection of fifty strains of Staphylococcus aureus isolated in Algeria at the university hospital of Telmcan. J Bacterio Rese 2014;6:1-6.

Mohamed A, Rajaa AM, Khalid Z, Fouad M, Naima R. Comparison of three methods for the detection of biofilm formation by clinical isolates of S. aureus isolated in Casablanca. Int J Sci Rese 2016;5:1156-9.

Hassan A, Usman J, Kaleen F, Omkar M, Khalid A, Iqbal M. Evaluation of different detection methods of biofilm formation in the clinical isolates. Braz J Infect Dis 2011;15:305-11.

Devaraj C, Sajjan GA. Comparison of three different methods for detection of biofilm in gram-positive cocci and gram-negative Bacilli isolated from clinical specimens. J Pharm Sci Rese 2015;7:952-5.

Grinholc M, Wegryn G, Kurlend J. Evaluation of biofilm production and prevalence of the icaD gene in methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains isolated from patients with nosocomial infections and carrier. FEMS Immun Med Microbiol 2007;50:375-9.