Author(s): Shaimaa Wahman, Mohamed Emara, Riham M. Shawky

Email(s): , ,

DOI: 10.52711/0974-360X.2023.00373   

Address: Shaimaa Wahman, Mohamed Emara, Riham M. Shawky
Department of Microbiology and Immunology, Faculty of Pharmacy, Helwan University, Ain Helwan, Helwan, 11795, Cairo, Egypt.
*Corresponding Author

Published In:   Volume - 16,      Issue - 5,     Year - 2023

Staphylococci have been implicated in chronic device-related infections due to their ability to form resistant biofilms on implanted medical devices. For a long time, two different mechanisms of biofilm formation were known in Staphylococcus spp., the ica-dependent biofilms in MSSA and CoNS and the ica-independent biofilms in MRSA. Recently, a new fibrin-based biofilm phenotype was identified when S. aureus isolates were allowed to construct biofilms in biologically-relevant conditions using plasma-coated surfaces and RPMI-1640 for biofilm development (RPMI-1640/Pl). In this study, 140 staphylococci clinical isolates (91 MRSA, 27 MSSA and 22 CoNS) were tested for biofilm formation, biofilm formers were selected and used to scrutinize the ability of RPMI-1640/Pl to support staphylococci biofilm formation. Results showed that, in RPMI-1640/Pl, the biofilm formation abilities of MRSA and MSSA isolates were non-significantly different compared to those formed in TSB and BHI, (Kruskal Wallis test, P = 0.3275 and 0.466 for MRSA and MSSA isolates, respectively). However, a significantly different biofilm formation ability was observed regarding the tested CoNS isolates (ANOVA test, P = 0.0006). Furthermore, biofilm formation in RPMI-1640/Pl under different incubation conditions was tested, and among the tested conditions, 48h of static incubation showed significantly elevated biofilm for both MRSA and MSSA. Finally, PCR was used to detect genes implicated in biofilm formation, and the genotypes were correlated to the biofilm formation ability in different tested conditions. In contrast to ordinary media, biofilm formation by staphylococci in RPMI-1640/Pl was positively correlated to coa, fnbA, fnbB and clfB.

Cite this article:
Shaimaa Wahman, Mohamed Emara, Riham M. Shawky. In-vitro assessment of staphylococci biofilms formed under biologically-relevant conditions and correlation to the biofilm genotype. Research Journal of Pharmacy and Technology 2023; 16(5):2273-9. doi: 10.52711/0974-360X.2023.00373

Shaimaa Wahman, Mohamed Emara, Riham M. Shawky. In-vitro assessment of staphylococci biofilms formed under biologically-relevant conditions and correlation to the biofilm genotype. Research Journal of Pharmacy and Technology 2023; 16(5):2273-9. doi: 10.52711/0974-360X.2023.00373   Available on:

1.    Umamageswari S, et al. Evaluation of antibacterial activity of zinc oxide nanoparticles against biofilm producing methicillin resistant Staphylococcus aureus (MRSA). Research Journal of Pharmacy and Technology. 2018; 11(5):1884-1888.10.5958/0974-360X.2018.00350.5
2.    Tong SY, et al. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clinical Microbiology Reviews. 2015; 28(3):603-661.
3.    Von Eiff C, et al. Pathogenesis of infections due to coagulase negative Staphylococci. The Lancet Infectious Diseases. 2002; 2(11):677-685.
4.    Trivedi U, et al. A post-planktonic era of in vitro infectious models: issues and changes addressed by a clinically relevant wound like media. Critical Reviews in Microbiology. 2017; 43(4):453-465.
5.    Idrees M, et al. Staphylococcus aureus biofilm: Morphology, genetics, pathogenesis and treatment strategies. International Journal of Environmental Research and Public Health. 2021; 18(14):7602.
6.    McCarthy H, et al. Methicillin resistance and the biofilm phenotype in Staphylococcus aureus. Frontiers in Cellular and Infection Microbiology. 2015; 5(1):1.
7.    Kranjec C, et al. Staphylococcal biofilms: Challenges and novel therapeutic perspectives. Antibiotics. 2021; 10(2):131.
8.    O'Neill E, et al. Association between methicillin susceptibility and biofilm regulation in Staphylococcus aureus isolates from device-related infections. Journal of Clinical Microbiology. 2007; 45(5):1379-1388.
9.    Zapotoczna M, et al. Untangling the diverse and redundant mechanisms of Staphylococcus aureus biofilm formation. PLoS Pathogens. 2016; 12(7):e1005671.
10.    Zapotoczna M, et al. An essential role for coagulase in Staphylococcus aureus biofilm development reveals new therapeutic possibilities for device-related infections. The Journal of Infectious Diseases. 2015; 212(12):1883-1893.
11.    Deepigaa M. Antibacterial resistance of bacteria in biofilms. Research Journal of Pharmacy and Technology. 2017; 10(11):4019-4023.10.5958/0974-360X.2017.00728.4
12.    Abbas HA, et al. Combating Pseudomonas aeruginosa biofilms by potential biofilm inhibitors. Asian J Res Pharm Sci. 2012; 2(2):66-72.
13.    Abbas HA, et al. Biofilms: The microbial castle of resistance. Research Journal of Pharmacy and Technology. 2013; 6(1):2.
14.    Seng R, et al. Biofilm formation of methicillin-resistant coagulase negative staphylococci (MR-CoNS) isolated from community and hospital Environments. PLoS One. 2017; 12(8):e0184172.
15.    Craft KM, et al. Methicillin-resistant Staphylococcus aureus (MRSA): antibiotic-resistance and the biofilm phenotype. Med Chem Comm. 2019; 10(8):1231-1241.10.1039/C9MD00044E
16.    Rammo RN. Bactericidal and anti-biofilm formation of aqueous plant extracts against pathogenic bacteria. Asian J. Pharm. Res. 2017; 7(1):25-29.10.5958/2231-5691.2017.00005.3
17.    Abdelghafar A, et al. Combating Staphylococcus aureus biofilm with Antibiofilm agents as an efficient strategy to control bacterial infection. Research Journal of Pharmacy and Technology. 2020; 13(11):5601-5606.10.5958/0974-360X.2020.00977.4
18.    Hamzah H, et al. Efficacy of quercetin against polymicrobial biofilm on catheters. Research Journal of Pharmacy and Technology. 2020; 13(11):5277-5282.10.5958/0974-360X.2020.00923.3
19.    Bhavani G. Detection of biofilm among clinical isolates of Acinetobacter baumannii by tissue culture plate method (TCP). Research Journal of Pharmacy and Technology. 2016; 9(10):1635-1637.10.5958/0974-360X.2016.00327.9
20.    Bostanghadiri N, et al. Antibiotic resistance, biofilm formation, and biofilm-associated genes among Stenotrophomonas maltophilia clinical isolates. BMC Research Notes. 2021; 14(1):1-6.
21.    Karimi K, et al. Investigation of antibiotic resistance and biofilm formation in clinical isolates of Klebsiella pneumoniae. International Journal of Microbiology. 2021; 2021(2021):5573388.
22.    Kishii K, et al. Differences in biofilm formation and transcription of biofilm-associated genes among Acinetobacter baumannii clinical strains belonging to the international clone II lineage. Journal of Infection and Chemotherapy. 2020; 26(7):693-698.
23.    Triveni AG, et al. Biofilm formation by clinically isolated Staphylococcus aureus from India. The Journal of Infection in Developing Countries. 2018; 12(12):1062-1066.
24.    Hamzah H, et al. The Inhibition and Degradation Activity of Demethoxycurcumin as Antibiofilm on C. albicans ATCC 10231. Research Journal of Pharmacy and Technology. 2020; 13(1):377-382.10.5958/0974-360X.2020.00075.X
25.    Shaaban M, et al. Antimicrobial and antibiofilm activities of probiotic lactobacilli on antibiotic-resistant proteus mirabilis. Microorganisms. 2020; 8(6):960.
26.    Qian W, et al. Antimicrobial mechanism of luteolin against Staphylococcus aureus and Listeria monocytogenes and its antibiofilm properties. Microbial Pathogenesis. 2020; 142(1):104056.
27.    Song YJ, et al. Anti-biofilm activity of grapefruit seed extract against Staphylococcus aureus and Escherichia coli. J. Microbiol. Biotechnol. 2019; 29(8):1177-1183.
28.    Wahman S, et al. Inhibition of quorum sensing‐mediated biofilm formation in Pseudomonas aeruginosa by a locally isolated Bacillus cereus. Journal of Basic Microbiology. 2015; 55(12):1406-1416.
29.    Bai J-R, et al. Antibiofilm activity of shikimic acid against Staphylococcus aureus. Food Control. 2019; 95(1):327-333.
30.    Mary R, Banu N. Inhibition of antibiofilm mediated virulence factors in Pseudomonas aeruginosa by Andrographis paniculata. Research Journal of Pharmacy and Technology. 2017; 10(1):141-144.10.5958/0974-360X.2017.00031.2
31.    Vaishali M, et al. Inhibitory Effect of Pomegranate Oil on Biofilm Formation–An In vitro Study. Research Journal of Pharmacy and Technology. 2018; 11(2):521-522.10.5958/0974-360X.2018.00096.3
32.    Kavaliauskas P, et al. Synthesis, ADMET properties, and in vitro antimicrobial and antibiofilm activity of 5-nitro-2-thiophenecarbaldehyde N-((E)-(5-nitrothienyl) methylidene) hydrazone (KTU-286) against Staphylococcus aureus with defined resistance mechanisms. Antibiotics. 2020; 9(9):612.
33.    Herrera K, et al. Antibacterial and antibiofilm activities of synthetic analogs of 3-alkylpyridine marine alkaloids. Medicinal Chemistry Research. 2020; 29(6):1084-1089.
34.    Baseri N, et al. Phenotypic and genotypic changes of Staphylococcus aureus in the presence of the inappropriate concentration of chlorhexidine gluconate. BMC Microbiology. 2022; 22(1):1-9.
35.    Forbes BA, et al. Overview of Bacterial Identification Methods and Strategies, In Bailey & Scott’s Diagnostic Microbiology, Edited by  Wilson L. 2007; 12th ed: pp. 216-248.
36.    Maes N, et al. Evaluation of a triplex PCR assay to discriminate Staphylococcus aureus from coagulase-negative staphylococci and determine methicillin resistance from blood cultures. Journal of Clinical Microbiology. 2002; 40(4):1514-1517.
37.    Himabindu M, et al. Molecular analysis of coagulase gene polymorphism in clinical isolates of methicillin resistant Staphylococcus aureus by restriction fragment length polymorphism based genotyping. Am J Infect Dis. 2009; 5(2):163-9.
38.    Goh S-H, et al. Molecular typing of Staphylococcus aureus on the basis of coagulase gene polymorphisms. Journal of Clinical Microbiology. 1992; 30(7):1642-1645.
39.    Javid F, et al. Molecular typing of Staphylococcus aureus based on coagulase gene. Veterinary World. 2018; 11(4):423.10.14202/vetworld.2018.423-430
40.    Atshan SS, et al. Prevalence of adhesion and regulation of biofilm-related genes in different clones of Staphylococcus aureus. Journal of Biomedicine and Biotechnology. 2012; 2012(2012):976972.
41.    Vancraeynest D, et al. Genotypic and phenotypic screening of high and low virulence Staphylococcus aureus isolates from rabbits for biofilm formation and MSCRAMMs. Veterinary Microbiology. 2004; 103(3-4):241-247.
42.    Rajesh P, Rai VR. Quorum quenching activity in cell-free lysate of endophytic bacteria isolated from Pterocarpus santalinus Linn., and its effect on quorum sensing regulated biofilm in Pseudomonas aeruginosa PAO1. Microbiological Research. 2014; 169(7-8):561-569.
43.    Achek R, et al. Phenotypic and molecular detection of biofilm formation in Staphylococcus aureus isolated from different sources in Algeria. Pathogens. 2020; 9(2):153.
44.    Omidi M, et al. Ability of biofilm production and molecular analysis of spa and ica genes among clinical isolates of methicillin-resistant Staphylococcus aureus. BMC Research Notes. 2020; 13(1):1-7.
45.    Uribe-García A, et al. Frequency and expression of genes involved in adhesion and biofilm formation in Staphylococcus aureus strains isolated from periodontal lesions. Journal of Microbiology, Immunology and Infection. 2021; 54(2):267-275.
46.    Petrova OE, Sauer K. Escaping the biofilm in more than one way: desorption, detachment or dispersion. Current Opinion in Microbiology. 2016; 30(1):67-78.

Recomonded Articles:

Research Journal of Pharmacy and Technology (RJPT) is an international, peer-reviewed, multidisciplinary journal.... Read more >>>

RNI: CHHENG00387/33/1/2008-TC                     
DOI: 10.5958/0974-360X 

56th percentile
Powered by  Scopus

SCImago Journal & Country Rank

Recent Articles


Not Available