Banat, Ibrahim M

Dooley, JSG

Lemoine, P

Naughton, Patrick

Puyol McKenna, Patricia

Ternan, Nigel G

English

Ulster University's Research Portal

                      Investigation of the anti-biofilm properties of purified biosurfactants in EnterococcalbiofilmsPuyol McKenna, P., Dooley, JSG., Ternan, N. G., Banat, I. M., Lemoine, P., & Naughton, P. (2023). Investigationof the anti-biofilm properties of purified biosurfactants in Enterococcal biofilms.Link to publication record in Ulster University Research PortalPublication Status:Published (in print/issue): 07/08/2023General rightsCopyright for the publications made accessible via Ulster University's Research Portal is retained by the author(s) and / or other copyrightowners and it is a condition of accessing these publications that users recognise and abide by the legal requirements associated with theserights.Take down policyThe Research Portal is Ulster University's institutional repository that provides access to Ulster's research outputs. Every effort has beenmade to ensure that content in the Research Portal does not infringe any person's rights, or applicable UK laws. If you discover content inthe Research Portal that you believe breaches copyright or violates any law, please contact pure-support@ulster.ac.uk.Download date: 14/09/2023•  Intro po•  Intro po•  Intro point 3 etc…Investigation of the anti-biofilm properties of purified biosurfactants in Enterococcal biofilmsPatricia Puyol McKenna1, James S.G. Dooley1, Nigel G. Ternan1, Ibrahim M. Banat1, Patrick Lemoine2, Patrick J. Naughton1.1Nutrition Innovation Centre for Food & Health (NICHE), Ulster University, Coleraine, BT52 1SA2School of Engineering, Nanotechnology and Integrated Bioengineering Centre (NIBEC), Ulster University, Belfast, United Kingdomulster.ac.uk/nicheFunding and acknowledgements. PPM was recipient of a PhD studentship awarded by Department for the Economy (DfE) NI. PPM acknowledges travel fundingfrom the Medical Research Foundation.@NICHE_ULSTER niche_ulster puyol_mckenna-p@ulster.com NICHEUlster /UlsterUniversityNICHEResultsadded to 24-well plates –incubate 24 hStain biofilms with 1 mL of 0.1 % crystal violetRemove crystal violet and wash twice with 1 mLof DI water – dry overnightDe-stain with 30 % acetic acid and measure at 570 nmStrain (E. faecalis)Mono Rhamnolipid(µg/mL)MIC MBCMW01105 250 500MF06036 250 500MW02048 250 500Aim/Objectives• To determine the inhibitory effects of 24-hour biosurfactant congener treatment (Mono Rhamnolipid) on 24, 48 and 72 hour E. faecalis biofilms.• To determine the biofilm disruption effects of the congeners on established 24, 48 and 72 hour E. faecalis biofilms.• To determine minimum inhibitory and biocidal concentrations of the congener by broth microdilution.ConclusionsMono-Rhamnolipid structureMinimum inhibitory and biocidal concentrations–determined by broth microdilution, growth measured by optical density at 600 nm.Biofilm inhibition Biofilm disruptionCultures added to 24-well plates + congener treatmentCultures added to 24-well  plates –incubate 24, 48, 72 hIncubation for 24, 48, 72 hCongener added to biofilms in replacement media-incubate 24 hMedia removed and plates washed twice with DI waterBiofilm staining with 1 mL of 0.1% crystal violet• Mono-RL shows significant (P<0.001) efficacy in biofilm inhibition and disruption. Results show a dose-dependent response, with higher concentrations of biosurfactants leading to greater inhibition/disruption of biofilm growth. • Preliminary results suggest that biosurfactants may offer a promising alternative or adjunctive treatment option against enterococcal biofilms. Fig 3. Biofilm biomass after co-incubation for 24, 48 and 72 h with Mono-RL. n = 9, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001.Fig 4. Biofilm biomass after 24, 48 and 72 h with a futher 24 h of Mono-RL treatment. n = 9, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001.Patricia_Mono_RL_PA_0_1mg_mL_neg_02Feb2023 #1-1607 RT: 0.01-40.09 AV: 1607 NL: 4.01E4T: - c ESI Full ms [ 110.00-2000.00]200 400 600 800 1000 1200 1400 1600 1800 2000m/z05101520253035404550556065707580859095100Relative Abundance503.0531.01006.8474.9 677.3649.0723.1212.0 451.2325.1 544.9 1021.4738.9 954.9903.6 1181.1205.0 1247.1 1533.11405.7 1633.6 1805.7 1864.4Fig 1. Rha-C10-C10. Molecular weight = 504.7 g/mol. Structure of RL consists of 2 β-hydroxyl fatty acids attached by a glycosidic linkage to 1 rhamnose sugar (PubChem, 2021). Mono-RL, Rha-C10-C10 (94%)Methods MIC –the minimum concentration at which there is a significant decrease in growth as measured by attenuance (D) at 600 nm.Introduction• Enterococcus spp. have become a leading causative of healthcare-associated infections (HAIs) ranging from bloodstream infections to UTIs1. Some 30% of healthcare-associated enterococcal infections are caused by vancomycin-resistant enterococci (VRE), requiring prolonged antibiotic therapy or becoming resistant to antibiotic monotherapy2.• Biofilms, structured biological communities that grow enveloped in an extracellular matrix of protective polysaccharides that subsequently prevents the action of antimicrobial agents3, can develop on an array of surfaces, from indwelling medical devices such as catheters to infected heart valves, facilitating chronic infections4.• Biosurfactants are unique amphiphilic molecules of microbial origin that are capable of interacting with the lipidic components of microorganisms5. Biosurfactant interactions  with different surfaces can affect their hydrophobic properties; their ability to alter microorganisms’ adhesion abilities and consequent biofilm formation6 could make biosurfactants suitable for targeted use in medical and pharmaceutical applications7.Table 1. MIC and MBC of Mono-RL for E. faecalis strains.1. Codelia-Anjum et al.,(2023). Enterococcal Urinary Tract Infections: A Review of the Pathogenicity, Epidemiology, and Treatment. Antibiotics.2. CDC (2019). Biggest Threats and Data. Centers for Disease Control and Prevention.3. Luo et al.,(2021). Mechanisms and Control Strategies of Antibiotic Resistance in Pathological Biofilms. Journal of Microbiology and Biotechnology4. Conwell et al.,(2022). Enterococcal biofilm – a nidus for antibiotic resistance transfer? Journal of Applied Microbiology. 5. Ceresa et al.,(2020). The effect of sophorolipids against microbial biofilms on medical-grade silicone. Journal of Biotechnology. 6. Kaczorek et al., (2018). The Impact of Biosurfactants on Microbial Cell Properties Leading to Hydrocarbon Bioavailability Increase.7. Ferreira et al., (2017). Novel cosmetic formulations containing a biosurfactant from Lactobacillus paracasei. Colloids and Surfaces B: BiointerfacesLC-MS/ESI mass spectrum in negative ion mode – purity 94%Purity of Mono-Rhamnolipid

Investigation of the anti-biofilm properties of purified biosurfactants in Enterococcal biofilms

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Investigation of the anti-biofilm properties of purified biosurfactants in Enterococcal biofilms

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