AN IN-VITRO ANTIBIOFILM ACTIVITY OF CHLORELLA VULGARIS

: Antibiofilm, Quorum sensing, freshwater algae, Chlorella vulgaris, Phytochemicals.

  • SRIDEVI NS Department of Biochemistry, Biotechnology and Bioinformatics, Avinashilingam Institute for Home Science and Higher Education for Women, Coimbatore, Tamil Nadu, India.
  • DHANUSHA V Department of Biochemistry, Biotechnology and Bioinformatics, Avinashilingam Institute for Home Science and Higher Education for Women, Coimbatore, Tamil Nadu, India.
  • RAJESWARI M Department of Biochemistry, Biotechnology and Bioinformatics, Avinashilingam Institute for Home Science and Higher Education for Women, Coimbatore, Tamil Nadu, India.
  • SANTHI N Department of Biochemistry, Biotechnology and Bioinformatics, Avinashilingam Institute for Home Science and Higher Education for Women, Coimbatore, Tamil Nadu, India.

Abstract

Objective: Most of the microbial infection in the body is through biofilm formation. Quorum sensing (QS) is the key regulator in the biofilm formation in both Gram-negative and Gram-positive bacteria. Therefore, interfering with QS is the current strategy to prevent bacterial infection.


Methods: In this study, the effect of various extracts of freshwater microalgae – Chlorella vulgaris on the growth of clinical pathogens – Pseudomonas aeruginosa and Staphylococcus aureus which were studied using minimum inhibitory concentration (MIC), antibiofilm activity, and (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay using 96-well flat bottom microtiter plates. The phytochemical analysis of C. vulgaris was also carried using standard procedure.


Results: The petroleum ether, dichloromethane, chloroform, ethyl acetate, and methanolic and acetone extract of C. vulgaris showed the presence of carbohydrates, amino acids, proteins, steroids, flavonoids, alkaloids, saponins, and phenolic compounds. The MIC value of methanolic extract of C. vulgaris was found to be 1 mg/ml. The highest inhibition of 82.5% and 88.0% was shown by methanolic extract at a concentration of 1 mg/ml for P. aeruginosa and S. aureus, respectively. The antibiofilm activity by crystal violet and MTT assay confirmed the reduction of biofilm formation in both pathogenic organisms.


Conclusion: The present results suggested the possible use of C. vulgaris in attenuation of QS and biofilm formation to control pathogenic bacteria – P. aeruginosa and S. aureus.

Keywords: Antibiofilm, Quorum sensing, Freshwater algae, Chlorella vulgaris, Phytochemicals

References

1. Costerton JW, Lewandowski Z, Caldwell DE, Korber DR, Lappin- Scott HM. Microbial biofilms. Annu Rev Microbiol 1995;49:711-45.
2. Kaufmann SH, Rouse BT, Sacks DL. The Immune Response to Infection. Washington, D.C: ASM Press; 2011.
3. Rossolini GM, Mantengoli E, Docquier JD, Musmanno RA, Coratza G. Epidemiology of infections caused by multiresistant gram-negatives: ESBLs, MBLs, panresistant strains. New Microbiol 2007;30:332-9.
4. Ghanbarzadeh Corehtash Z, Khorshidi A, Firoozeh F, Akbari H, Mahmoudi Aznaveh A. Biofilm formation and virulence factors among Pseudomonas aeruginosa isolated from burn patients. Jundishapur J Microbiol 2015;8:e22345.
5. Eisenstein BI. Treatment of staphylococcal infections with cyclic lipopeptides. Clin Microbiol Infect 2008;14 Suppl 2:10-6.
6. Ganesh PS, Rai VR. Attenuation of quorum-sensing-dependent virulence factors and biofilm formation by medicinal plants against antibiotic resistant Pseudomonas aeruginosa. J Tradit Complement Med 2018;8:170-7.
7. Kannappan A, Gowrishankar S, Srinivasan R, Pandian SK, Ravi AV. Antibiofilm activity of Vetiveria zizanioides root extract against methicillin-resistant Staphylococcus aureus. Microb Pathog 2017;110:313-24.
8. Sultana T, Mitra AK, Das S. A preliminary observation on an explicit antimicrobial action of mangrove plants on Pseudomonas aeruginosa. Asian J Pharm Clin Res 2019;12 Suppl 5:226-30.
9. Wu S, Liu G, Jin W, Xiu P, Sun C. Antibiofilm and anti-infection of a marine bacterial exopolysaccharide against Pseudomonas aeruginosa. Front Microbiol 2016;7:102.
10. Roy R, Tiwari M, Donelli G, Tiwari V. Strategies for combating bacterial biofilms: A focus on anti-biofilm agents and their mechanisms of action. Virulence 2018;9:522-54.
11. Jamal M, Ahmad W, Andleeb S, Jalil F, Imran M, Nawaz MA, et al. Bacterial biofilm and associated infections. J Chin Med Assoc 2018;81:7-11.
12. Krishnan T, Yin WF, Chan KG. Inhibition of quorum sensing-controlled virulence factor production in Pseudomonas aeruginosa PAO1 by ayurveda spice clove (Syzygium aromaticum) bud extract. Sensors (Basel) 2012;12:4016-30.
13. Latifi A, Foglino M, Tanaka K, Williams P, Lazdunski A. A hierarchical quorum-sensing cascade in Pseudomonas aeruginosa links the transcriptional activators lasR and rhIR (VsmR) to expression of the stationary-phase sigma factor rpoS. Mol Microbiol 1996;21:1137-46.
14. Asfour HZ. Anti-quorum sensing natural compounds. J Microsc Ultrastruct 2018;6:1-10.
15. Sánchez E, Morales CR, Castillo S, Leos-Rivas C, García-Becerra L, Martínez DM. Antibacterial and antibiofilm activity of methanolic plant extracts against nosocomial microorganisms. Evid Based Complement Alternat Med 2016;2016:8.
16. Koo H, Allan RN, Howlin RP, Stoodley P, Hall-Stoodley L. Targeting microbial biofilms: Current and prospective therapeutic strategies. Nat Rev Microbiol 2017;15:740-55.
17. Kim SK, Ravichandran DY, Khan SB, Kim YT. Prospective of the cosmeceuticals derived from marine organisms. Biotechnol Bioprocess Eng 2008;13 Suppl 5:511-23.
18. Wang HM, Pan JL, Chen CY, Chiu CC, Yang MH, Chang HW, et al. Identification of anti-lung cancer extract from Chlorella vulgaris C-C by antioxidant property using supercritical carbon dioxide extraction. Process Biochem 2010;45 Suppl 12:1865-72.
19. Harborne JB. Phytochemical Methods. 3rd ed. London: Chapman and Hall; 1998.
20. Wiegand I, Hilpert K, Hancock RE. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc 2008;3:163-75.
21. Stepanovic S, Vukovic D, Dakic I, Savic B, Svabic-Vlahovic M. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J Microbiol Methods 2000;40:175-9.
22. Schillaci D, Cusimano MG, Cunsolo V, Saletti R, Russo D, Vazzana M, et al. Immune mediators of sea-cucumber Holothuria tubulosa (Echinodermata) as source of novel antimicrobial and anti-staphylococcal biofilm agents. AMB Express 2013;3:35.
23. Nemati F, Dehpouri AA, Eslami B, Mahdavi V, Mirzanejad S. Cytotoxic properties of some medicinal plant extracts from mazandaran, iran. Iran Red Crescent Med J 2013;15:e8871.
24. Annamalai J, Nallamuthu T. Antioxidant potential phytochemicals from methanol extract of Chlorella vulgaris and Chlamydomonas reinhardtii. J Algal Biomass Util 2014;5 Suppl 4:60-7.
25. Naves P, del Prado G, Huelves L, Gracia M, Ruiz V, Blanco J, et al. Measurement of biofilm formation by clinical isolates of Escherichia coli is method-dependent. J Appl Microbiol 2008;105:585-90.
26. Mutungwa M, Alluri N, Majumdar M. Anti-quorum sensing activity of some commonly used traditional Indian spices. Int J Pharm Pharm Sci 2015;7 Suppl 11:80-3.
27. Pratiwi SU, Lagengijk EL, Hertiani T, Weert SD, Cornellius AM, Hondel JJ. Antimicrobial effects of Indonesian medicinal plants extract on planktonic and biofilm growth of Pseudomonas aeruginosa and Staphylococcus aureus. Int J Pharm Pharm Sci 2015;7 Suppl 4:183-91.
28. Gayatri KV, Soundhari C, Pavithra BP. Biofilm inhibitory effect of Chlorella extracts on Pseudomonas aeruginosa. Int J Pharm Sci Res 2019;10 Suppl 4:1966-71.
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SRIDEVI NS, DHANUSHA V, RAJESWARI M, and SANTHI N. “AN IN-VITRO ANTIBIOFILM ACTIVITY OF CHLORELLA VULGARIS”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 12, no. 8, July 2019, pp. 239-42, https://innovareacademics.in/journals/index.php/ajpcr/article/view/34144.
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