Abstract
This study aimed to investigate the ability of twelve strains of Enterococcus spp. to form biofilms on stainless steel (AISI 316 L and AISI 304 L) and to characterize the physicochemical properties of the bacterial and surface supports. The study also examined the auto-aggregation and extracellular polymeric substances of strains in both planktonic and sessile forms. The results showed that all Enterococcus spp. strains tested were capable of forming biofilms, with varying degrees reaching 7.41 log CFU/cm2 on AISI 316 L for Enterococcus mundtii A1. On AISI 304 L, the highest biofilm recorded was 7.01 log CFU/cm2 for Enterococcus mundtii A13. It was observed that auto-aggregation was influenced by the strains’ hydrophobicity. These results shed light on the possible application of Enterococcus spp. biofilm as a strategy to control pathogenic biofilms in various industries and provide insight into the role of physicochemical properties in biofilm formation.
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Abbreviations
- LAB:
-
Lactic acid bacteria
- GRAS:
-
Generally recognized as safe
- MRS:
-
Man Rogosa and Sharpe
- CFU:
-
Colony forming units
- EPS:
-
Extracellular polymeric substances
- PCA:
-
Principal component analysis
References
Ait Ouali F, Al Kassaa I, Cudennec B, Abdallah M, Bendali F, Sadoun D, Chihib N-E, Drider D (2014) Identification of lactobacilli with inhibitory effect on biofilm formation by pathogenic bacteria on stainless steel surfaces. Int J Food Microbiol 191:116–124. https://doi.org/10.1016/j.ijfoodmicro.2014.09.011
Al-Hamarneh I, Pedrow P, Eskhan A, Abu-Lail N (2012) Hydrophilic property of 316L stainless steel after treatment by atmospheric pressure corona streamer plasma using surface-sensitive analyses. Appl Surf Sci 259:424–432. https://doi.org/10.1016/j.apsusc.2012.07.061
Amaral DMF, Silva LF, Casarotti SN, Nascimento LCS, Penna ALB (2017) Enterococcus faecium and Enterococcus durans isolated from cheese: survival in the presence of medications under simulated gastrointestinal conditions and adhesion properties. J Dairy Sci 100(2):933–949. https://doi.org/10.3168/jds.2016-11513
Angmo K, Kumari A, Savitri, Bhalla TC (2016) Probiotic characterization of lactic acid bacteria isolated from fermented foods and beverage of Ladakh. LWT 66:428–435. https://doi.org/10.1016/j.lwt.2015.10.057
Asria M, Ouafib R, Bahafida W, Elabeda S, Koraichia SI, Costad F, Tavaresd T, El Ghachtoulia N (2023) Chromium removal by newly developed microbial consortia supported on wood husk. Desalination Water Treat 289:80–91. https://doi.org/10.5004/dwt.2023.29342
Avram-Hananel L, Stock J, Parlesak A, Bode C, Schwartz B (2010) E Durans strain M4–5 isolated from human colonic Flora attenuates intestinal inflammation. Dis Colon Rectum 53(12):1676–1686. https://doi.org/10.1007/DCR.0b013e3181f4b148
Ayyash M, Abushelaibi A, Al-Mahadin S, Enan M, El-Tarabily K, Shah N (2018) In-vitro investigation into probiotic characterisation of Streptococcus and Enterococcus isolated from camel milk. LWT 87:478–487. https://doi.org/10.1016/j.lwt.2017.09.019
Azelmad K, Hamadi F, Mimouni R, Amzil K, Latrache H, Mabrouki M, El Boulani A (2017) Adhesion of Staphylococcus aureus and staphylococcus xylosus to materials commonly found in catering and domestic kitchens. Food Control 73:156–163. https://doi.org/10.1016/j.foodcont.2016.07.044
Bohinc K, Dražić G, Fink R, Oder M, Jevšnik M, Nipič D, Godič-Torkar K, Raspor P (2014) Available surface dictates microbial adhesion capacity. Int J Adhes Adhes 50:265–272. https://doi.org/10.1016/j.ijadhadh.2014.01.027
Botes M, Loos B, van Reenen CA, Dicks LMT (2008) Adhesion of the probiotic strains Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 to Caco-2 cells under conditions simulating the intestinal tract, and in the presence of antibiotics and anti-inflammatory medicaments. Arch Microbiol 190(5):573–584. https://doi.org/10.1007/s00203-008-0408-0
Busscher HJ, Weerkamp AH, van der Mei HC, van Pelt AW, de Jong HP, Arends J (1984) Measurement of the surface free energy of bacterial cell surfaces and its relevance for adhesion. Appl Environ Microbiol 48(5):980–983. https://doi.org/10.1128/aem.48.5.980-983.1984
Casarin LS, Casarin FO, Brandelli A, Novello J, Ferreira SO, Tondo EC (2016) Influence of free energy on the attachment of Salmonella enteritidis and Listeria monocytogenes on stainless steels AISI 304 and AISI 316. LWT 69:131–138. https://doi.org/10.1016/j.lwt.2016.01.035
Castillo Pedraza MC, Novais TF, Faustoferri RC, Quivey RG, Terekhov A, Hamaker BR, Klein MI (2017) Extracellular DNA and lipoteichoic acids interact with exopolysaccharides in the extracellular matrix of Streptococcus mutans biofilms. Biofouling 33(9):722–740. https://doi.org/10.1080/08927014.2017.1361412
Chae MS, Schraft H, Truelstrup Hansen L, Mackereth R (2006) Effects of physicochemical surface characteristics of Listeria monocytogenes strains on attachment to glass. Food Microbiol 23(3):250–259. https://doi.org/10.1016/j.fm.2005.04.004
Costa OYA, Raaijmakers JM, Kuramae EE (2018) Microbial extracellular polymeric substances: ecological function and impact on soil aggregation. Front Microbiol. https://doi.org/10.3389/fmicb.2018.01636
Dertli E, Mayer MJ, Narbad A (2015) Impact of the exopolysaccharide layer on biofilms, adhesion and resistance to stress in Lactobacillus johnsonii FI9785. BMC Microbiol 15(1):8. https://doi.org/10.1186/s12866-015-0347-2
Du R, Ping W, Ge J (2022) Purification, characterization and mechanism of action of enterocin HDX-2, a novel class IIa bacteriocin produced by Enterococcus faecium HDX-2. LWT 153:112451. https://doi.org/10.1016/j.lwt.2021.112451
DuBois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28(3):350–356. https://doi.org/10.1021/ac60111a017
Ehrmann MA, Kurzak P, Bauer J, Vogel RF (2002) Characterization of lactobacilli towards their use as probiotic adjuncts in poultry. J Appl Microbiol 92(5):966–975. https://doi.org/10.1046/j.1365-2672.2002.01608.x
Elfazazi K, Zahir H, Tankiouine S, Mayoussi B, Zanane C, Lekchiri S, Ellouali M, Mliji EM, Latrache H (2021) Adhesion behavior of Escherichia coli strains on glass: role of cell surface qualitative and quantitative hydrophobicity in their attachment ability. Int J Microbiol 5580274. https://doi.org/10.1155/2021/5580274
Faten K, Hamida K, Soumya EA, Saad ISK, Hasna M, Hassan L, Moktar H (2016) Lactobacillus plantarum: effect of a protective biofilm on the surface of olives during storage. Braz J Microbiol 47. https://doi.org/10.1016/j.bjm.2015.11.028
Favaro L, Basaglia M, Casella S, Hue I, Dousset X, de Melo Franco BDG, Todorov SD (2014) Bacteriocinogenic potential and safety evaluation of non-starter Enterococcus faecium strains isolated from home made white brine cheese. Food Microbiol 38:228–239. https://doi.org/10.1016/j.fm.2013.09.008
Flint SH, Bremer PJ, Brooks JD (1997) Biofilms in dairy manufacturing plant-description, current concerns and methods of control. Biofouling 11(1):81–97. https://doi.org/10.1080/08927019709378321
Franz CMAP, Huch M, Abriouel H, Holzapfel W, Gálvez A (2011) Enterococci as probiotics and their implications in food safety. Int J Food Microbiol 151(2):125–140. https://doi.org/10.1016/j.ijfoodmicro.2011.08.014
Ghosh A, Borst L, Stephen SH, Suyemoto M, Moisan P, Zurek L, Jody GL (2013) Mortality in kittens is associated with a shift in ileum mucosa-associated enterococci from Enterococcus hirae to biofilm-forming Enterococcus faecalis and adherent Escherichia coli. J Clin Microbiol 51(11):3567–3578. https://doi.org/10.1128/JCM.00481-13
Giaouris E, Chapot-Chartier MP, Briandet R (2009) Surface physicochemical analysis of natural Lactococcus lactis strains reveals the existence of hydrophobic and low charged strains with altered adhesive properties. Int J Food Microbiol 131(1):2–9. https://doi.org/10.1016/j.ijfoodmicro.2008.09.006
Giraffa G, De Fernando GG (2022) Lactic acid bacteria: Enterococcus in milk and dairy products. In Reference Module in Food Science Elsevier BV. Amsterdam, The Netherlands, pp. 1–7
Gómez NC, Ramiro JMP, Quecan BXV, de Melo Franco BDG (2016) Use of potential probiotic lactic acid Bacteria (LAB) biofilms for the control of Listeria monocytogenes, salmonella typhimurium, and Escherichia coli O157:H7 biofilms formation. Front Microbiol 7. https://doi.org/10.3389/fmicb.2016.00863
Hamadi F, Latrache H, Mabrrouki M, Elghmari A, Outzourhit A, Ellouali M, Chtaini A (2005) Effect of pH on distribution and adhesion of Staphylococcus aureus to glass. J Adhes Sci Technol 19(1):73–85. https://doi.org/10.1163/1568561053066891
Hamadi F, Asserne F, Elabed S, Bensouda S, Mabrouki M, Latrache H (2014) Adhesion of Staphylococcus aureus on stainless steel treated with three types of milk. Food Control 38:104–108. https://doi.org/10.1016/j.foodcont.2013.10.006
Harimawan A, Ting Y-P (2016) Investigation of extracellular polymeric substances (EPS) properties of P. Aeruginosa and B. Subtilis and their role in bacterial adhesion. Colloids Surf B 146:459–467. https://doi.org/10.1016/j.colsurfb.2016.06.039
Jiang Z, Nero T, Mukherjee S, Olson R, Yan J (2021) Searching for the secret of stickiness: how biofilms adhere to surfaces. Front Microbiol 12. https://doi.org/10.3389/fmicb.2021.686793
Khalil AL, Oleiwi SR (2021) Effect of Lactobacillus fermentum filtrate on Pseudomonas aeruginosa adhesion at biotic and abiotic surfaces. Sys Rev Pharm 11(8):713–720
Kiani A, Nami Y, Hedayati S, Jaymand M, Samadian H, Haghshenas B (2021) Tarkhineh as a new microencapsulation matrix improves the quality and sensory characteristics of probiotic Lactococcus lactis KUMS-T18 enriched potato chips. Sci Rep 11(1):12599. https://doi.org/10.1038/s41598-021-92095-1
Kouider N, Hamadi F, Mallouki B, Bengourram J, Mabrouki M, Zekraoui M, Ellouali M, Latrache H (2010) Effect of stainless steel surface roughness on Staphylococcus aureus adhesion. Int j pure appl sci 4(1):1–7
Krausova G, Hyrslova I, Hynstova I (2019) In vitro evaluation of adhesion capacity, hydrophobicity, and auto-aggregation of newly isolated potential probiotic strains. Ferment 5(4):100. https://doi.org/10.3390/fermentation5040100
Li W, Ji J, Rui X, Yu J, Tang W, Chen X, Jiang M, Dong M (2014) Production of exopolysaccharides by Lactobacillus helveticus MB2-1 and its functional characteristics in vitro. LWT 59(2, Part 1):732–739. https://doi.org/10.1016/j.lwt.2014.06.063
Li Q, Liu X, Dong M, Zhou J, Wang Y (2015) Aggregation and adhesion abilities of 18 lactic acid bacteria strains isolated from traditional fermented food. Int J Agric Policy Res 3(2):84–92. https://doi.org/10.15739/IJAPR.030
Li W, Zhang J, Zhu Y, Li F (2021) Probiotic characterization of Enterococcus mundtii isolated from larval gut of the diamondback moth, Plutella xylostella. J Kans Entomol Soc 93(3):196–210. https://doi.org/10.2317/0022-8567-93.3.196
Liu H, Zhang L, Yi H, Han X, Gao W, Chi C, Song W, Li H, Liu C (2016) A novel enterocin T1 with anti-Pseudomonas activity produced by Enterococcus faecium T1 from Chinese Tibet cheese. World J Microbiol Biotechnol 32(2):21. https://doi.org/10.1007/s11274-015-1973-4
Ly MH, Vo NH, Le TM, Belin JM, Waché Y (2006) Diversity of the surface properties of Lactococci and consequences on adhesion to food components. Colloids and Surf B 52(2):149–153. https://doi.org/10.1016/j.colsurfb.2006.04.015
Meroufel AA (2020) Corrosion in reverse osmosis desalination processes: forms and mitigation practices. In: (eds) Corrosion and fouling control in desalination industry. Springer International Publishing, Cham, pp 131–151
Nami Y, Bakhshayesh RV, Jalaly HM, Lotfi H, Eslami S, Hejazi MA (2019) Probiotic Properties of Enterococcus Isolated From Artisanal Dairy Products Front Microbiol:10. https://doi.org/10.3389/fmicb.2019.00300
Nel HA, Bauer R, Wolfaardt GM, Dicks LMT (2002) Effect of Bacteriocins Pediocin PD-1, Plantaricin 423, and Nisin on biofilms of Oenococcus oeni on a stainless steel surface. Am J Enol Vitic 53(3):191. https://doi.org/10.5344/ajev.2002.53.3.191
Nikolic M, Jovcic B, Kojic M, Topisirovic L (2010) Surface properties of Lactobacillus and Leuconostoc isolates from homemade cheeses showing auto-aggregation ability. Eur Food Res Technol 231(6):925–931. https://doi.org/10.1007/s00217-010-1344-1
Nwoko EQA, Okeke IN (2021) Bacteria autoaggregation: how and why bacteria stick together. Biochem Soc Trans 49(3):1147–1157. https://doi.org/10.1042/BST20200718
Pérez Ibarreche M, Castellano P, Vignolo G (2014) Evaluation of anti-Listeria meat borne Lactobacillus for biofilm formation on selected abiotic surfaces. Meat Sci 96(1):295–303. https://doi.org/10.1016/j.meatsci.2013.07.010
Pérez Ibarreche M, Castellano P, Leclercq A, Vignolo G (2016) Control of Listeria monocytogenes biofilms on industrial surfaces by the bacteriocin-producing Lactobacillus sakei CRL1862. FEMS Microbiol Lett 363(12):118. https://doi.org/10.1093/femsle/fnw118
Pompilio A, Piccolomini R, Picciani C, D'Antonio D, Savini V, Di Bonaventura G (2008) Factors associated with adherence to and biofilm formation on polystyrene by Stenotrophomonas maltophilia: the role of cell surface hydrophobicity and motility. FEMS Microbiol Lett 287(1):41–47. https://doi.org/10.1111/j.1574-6968.2008.01292.x
Puniya M, Ravinder Kumar M, Panwar H, Kumar N, Ramneek AKP (2016) Screening of lactic acid bacteria of different origin for their probiotic potential. J Food Process Technol 7(1):545
Qiao X, Du R, Wang Y, Han Y, Zhou Z (2020) Purification, characterization and mode of action of enterocin, a novel bacteriocin produced by Enterococcus faecium TJUQ1. Int J Biol Macromol 144:151–159. https://doi.org/10.1016/j.ijbiomac.2019.12.090
Rodrigues L, van der Mei H, Teixeira JA, Oliveira R (2004) Biosurfactant from Lactococcus lactis 53 inhibits microbial adhesion on silicone rubber. Appl Microbiol Biotechnol 66(3):306–311. https://doi.org/10.1007/s00253-004-1674-7
Różańska A, Chmielarczyk A, Romaniszyn D, Sroka-Oleksiak A, Bulanda M, Walkowicz M, Osuch P, Knych T (2017) Antimicrobial properties of selected copper alloys on Staphylococcus aureus and Escherichia coli in different simulations of environmental conditions: with vs. without organic contamination. Int J Environ Res Public Health 14(7):813. https://doi.org/10.3390/ijerph14070813
Shuai C, Zhou Y, Yang Y, Gao C, Peng S, Wang G (2018) Ag-introduced antibacterial ability and corrosion resistance for bio-mg alloys. Biomed Res Int 6023460. https://doi.org/10.1155/2018/6023460
Sleytr UB (1978) Regular arrays of macromolecules on bacterial cell walls: structure, chemistry, assembly, and function. Int Rev Cytol 53:1–64. https://doi.org/10.1016/S0074-7696(08)62240-8
Sorroche FG, Spesia MB, Zorreguieta Á, Giordano W (2012) A positive correlation between bacterial autoaggregation and biofilm formation in native Sinorhizobium meliloti isolates from Argentina. Appl Environ Microbiol 78(12):4092–4101. https://doi.org/10.1128/AEM.07826-11
Speranza B, Sinigaglia M, Corbo MR (2009) Non starter lactic acid bacteria biofilms: a means to control the growth of Listeria monocytogenes in soft cheese. Food Control 20(11):1063–1067. https://doi.org/10.1016/j.foodcont.2009.01.006
Srivastava S, Bhargava A (2016) Biofilms and human health. Biotechnol Lett 38(1):1–22. https://doi.org/10.1007/s10529-015-1960-8
Tantratian S, Srimangkornkaew N, Prakitchaiwattana C, Sanguandeekul R (2022) Effect of different stainless steel surfaces on the formation and control of Vibrio parahaemolyticus biofilm. LWT 166:113788. https://doi.org/10.1016/j.lwt.2022.113788
Tran TTT, Kannoorpatti K, Padovan A, Thennadil S (2021) A study of bacteria adhesion and microbial corrosion on different stainless steels in environment containing Desulfovibrio vulgaris. R Soc Open Sci 8(1):201577. https://doi.org/10.1098/rsos.201577
Trunk T, Khalil HS, Leo JC (2018) Bacterial autoaggregation. AIMS Microbiol 4(1):140–164. https://doi.org/10.3934/microbiol.2018.1.140
Tuo Y, Yu H, Ai L, Wu Z, Guo B, Chen W (2013) Aggregation and adhesion properties of 22 Lactobacillus strains. J Dairy Sci 96(7):4252–4257. https://doi.org/10.3168/jds.2013-6547
Van Oss CJ, Chaudhury MK, Good RJ (1988) Interfacial Lifshitz-van der Waals and polar interactions in macroscopic systems. Chem Rev 88(6):927–941. https://doi.org/10.1021/cr00088a006
van Zyl WF, Deane SM, Dicks LMT (2016) Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 excludes Listeria monocytogenes from the GIT, as shown by bioluminescent studies in mice. Benef Microbes 7(2):227–235. https://doi.org/10.3920/BM2015.0082
Verran J, Rowe DL, Cole D, Boyd RD (2000) The use of the atomic force microscope to visualise and measure wear of food contact surfaces. Int Biodeterior Biodegrad 46(2):99–105. https://doi.org/10.1016/S0964-8305(00)00070-6
Xu H, Jeong HS, Lee HY, Ahn J (2009) Assessment of cell surface properties and adhesion potential of selected probiotic strains. Lett Appl Microbiol 49(4):434–442. https://doi.org/10.1111/j.1472-765X.2009.02684.x
Yin W, Wang Y, Liu L, He J (2019) Biofilms: the microbial “protective clothing” in extreme environments. Int J Mol Sci 20(14):3423. https://doi.org/10.3390/ijms20143423
Zanzan M, Achemchem F, Hamadi F, Latrache H, Elmoslih A, Mimouni R (2023) Anti-adherence activity of monomicrobial and polymicrobial food-derived Enterococcus spp. biofilms against pathogenic bacteria. Curr Microbiol 80:216. https://doi.org/10.1007/s00284-023-03326-9
Zhao Q, Liu Y, Wang C, Wang S, Peng N, Jeynes C (2007) Bacterial adhesion on ion-implanted stainless steel surfaces. Appl Surf Sci 253(21):8674–8681. https://doi.org/10.1016/j.apsusc.2007.04.044
Zhao T, Podtburg TC, Zhao P, Chen D, Baker DA, Cords B, Doyle MP (2013) Reduction by competitive bacteria of Listeria monocytogenes in biofilms and Listeria bacteria in floor drains in a ready-to-eat poultry processing plant. J Food Prot 76(4):601–607. https://doi.org/10.4315/0362-028X.JFP-12-323
Zommiti M, Cambronel M, Maillot O, Barreau M, Sebei K, Feuilloley M, Ferchichi M, Connil N (2018) Evaluation of probiotic properties and safety of Enterococcus faecium isolated from artisanal Tunisian meat “Dried Ossban”. Front Microbiol 9. https://doi.org/10.3389/fmicb.2018.01685
Acknowledgments
The authors would like to express their gratitude to Mr. Mouhamed BOUROUACH from the Faculty of Science in Agadir for his valuable assistance with principal component analysis. We would also like to thank El Mostafa Hamadi for his meticulous proofreading and editing of this manuscript.
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Zanzan, M., Ezzaky, Y., Achemchem, F. et al. Bacterial biofilm formation on stainless steel: exploring the capacity of Enterococcus spp. isolated from dairy products on AISI 316 L and AISI 304 L surfaces. Biologia (2024). https://doi.org/10.1007/s11756-024-01630-8
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DOI: https://doi.org/10.1007/s11756-024-01630-8