1887

Abstract

In natural habitats, bacterial species often coexist in biofilms. They interact in synergetic or antagonistic ways and their interactions can influence the biofilm development and properties. Still, very little is known about how the coexistence of multiple organisms impact the multispecies biofilm properties. In this study, we examined the behaviour of a dual-species biofilm at the air–liquid interface composed by two environmental bacteria: and a phenazine mutant of . Study of the planktonic and biofilm growths for each species revealed that grew faster than and no bactericidal effect from was detected, suggesting that the growth kinetics could be the main factor in the dual-species biofilm composition. To validate this hypothesis, the single- and dual-species biofilm were characterized by biomass quantification, microscopy and rheology. Bacterial counts and microscale architecture analysis showed that both bacterial populations coexist in the mature pellicle, with a dominance of . Real-time measurement of the dual-species biofilms' viscoelastic (i.e. mechanical) properties using interfacial rheology confirmed that was the main contributor of the biofilm properties. Evaluation of the dual-species pellicle viscoelasticity at longer time revealed that the biofilm, after reaching a first equilibrium, created a stronger and more cohesive network. Interfacial rheology proves to be a unique quantitative technique, which combined with microscale imaging, contributes to the understanding of the time-dependent properties within a polymicrobial community at various stages of biofilm development. This work demonstrates the importance of growth kinetics in the bacteria competition for the interface in a model dual-species biofilm.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000819
2019-07-01
2024-12-01
Loading full text...

Full text loading...

/deliver/fulltext/micro/165/7/747.html?itemId=/content/journal/micro/10.1099/mic.0.000819&mimeType=html&fmt=ahah

References

  1. Flemming HC, Wingender J, Szewzyk U, Steinberg P, Rice SA et al. Biofilms: an emergent form of bacterial life. Nat Rev Microbiol 2016; 14:563–575 [View Article]
    [Google Scholar]
  2. Høiby N. Recent advances in the treatment of Pseudomonas aeruginosa infections in cystic fibrosis. BMC Med 2011; 9:32 [View Article]
    [Google Scholar]
  3. Høiby N, Bjarnsholt T, Givskov M, Molin S, Ciofu O. Antibiotic resistance of bacterial biofilms. Int J Antimicrob Agents 2010; 35:322–332 [View Article]
    [Google Scholar]
  4. Wu H, Moser C, Wang HZ, Høiby N, Song ZJ. Strategies for combating bacterial biofilm infections. Int J Oral Sci 2015; 7:1–7 [View Article]
    [Google Scholar]
  5. Donlan RM, Rodney MD. Biofilms and device-associated infections. Emerg Infect Dis 2001; 7:277–281 [View Article]
    [Google Scholar]
  6. Percival SL, Hill KE, Williams DW, Hooper SJ, Thomas DW et al. A review of the scientific evidence for biofilms in wounds. Wound Repair Regen 2012; 20:647–657 [View Article]
    [Google Scholar]
  7. Høiby N, Ciofu O, Bjarnsholt T. Pseudomonas aeruginosa biofilms in cystic fibrosis. Future Microbiol 2010; 5:1663–1674 [View Article]
    [Google Scholar]
  8. Lazarova V, Perera J, Bowen M, Sheilds P. Application of aerated biofilters for production of high quality water for industrial reuse in West Basin. Water Sci Technol 2000; 41:417–424 [View Article]
    [Google Scholar]
  9. Majumder PS, Gupta SK. Hybrid reactor for priority pollutant nitrobenzene removal. Water Res 2003; 37:4331–4336 [View Article]
    [Google Scholar]
  10. Ho KG, Pometto AI, Hinz PN, Demirci A. Nutrient leaching and end product accumulation in plastic composite supports for L-(+)-lactic Acid biofilm fermentation. Appl Environ Microbiol 1997; 63:2524–2532
    [Google Scholar]
  11. Demirci A, Pometto AL, Ho KL. Ethanol production by Saccharomyces cerevisiae in biofilm reactors. J Ind Microbiol Biotechnol 1997; 19:299–304 [View Article]
    [Google Scholar]
  12. Branda SS, González-Pastor JE, Ben-Yehuda S, Losick R, Kolter R. Fruiting body formation by Bacillus subtilis . Proc Natl Acad Sci U S A 2001; 98:11621–11626 [View Article]
    [Google Scholar]
  13. Vlamakis H, Chai Y, Beauregard P, Losick R, Kolter R. Sticking together: building a biofilm the Bacillus subtilis way. Nat Rev Microbiol 2013; 11:157–168 [View Article]
    [Google Scholar]
  14. Branda SS, Vik Å, Friedman L, Kolter R. Biofilms: the matrix revisited. Trends in Microbiology 2005; 13:20–26 [View Article]
    [Google Scholar]
  15. Marvasi M, Visscher PT, Casillas Martinez L. Exopolymeric substances (Eps) from Bacillus subtilis: polymers and genes encoding their synthesis. FEMS Microbiol Lett 2010; 313:1–9 [View Article]
    [Google Scholar]
  16. Cutting SM. Bacillus probiotics. Food Microbiol 2011; 28:214–220 [View Article]
    [Google Scholar]
  17. Pasvolsky R, Zakin V, Ostrova I, Shemesh M. Butyric acid released during milk lipolysis triggers biofilm formation of Bacillus species. Int J Food Microbiol 2014; 181:19–27 [View Article]
    [Google Scholar]
  18. Kalogridou-Vassiliadou D. Biochemical activities of Bacillus species isolated from flat sour evaporated milk. J Dairy Sci 1992; 75:2681–2686 [View Article]
    [Google Scholar]
  19. Randrianjatovo-Gbalou I, Rouquette P, Lefebvre D, Girbal-Neuhauser E, Marcato-Romain CE. In situ analysis of Bacillus licheniformis biofilms: amyloid-like polymers and eDNA are involved in the adherence and aggregation of the extracellular matrix. J Appl Microbiol 2017; 122:1262–1274 [View Article]
    [Google Scholar]
  20. Baum MM, Kainović A, O'Keeffe T, Pandita R, McDonald K et al. Characterization of structures in biofilms formed by a Pseudomonas fluorescens isolated from soil. BMC Microbiol 2009; 9:103 [View Article]
    [Google Scholar]
  21. Rossi C, Chaves-López C, Serio A, Goffredo E, Goga BTC et al. Influence of incubation conditions on biofilm formation by Pseudomonas Fluorescens isolated from dairy products and dairy manufacturing plants. Ital J Food Saf 2016; 5:5793 [View Article]
    [Google Scholar]
  22. Spiers AJ, Bohannon J, Gehrig SM, Rainey PB. Biofilm formation at the air-liquid interface by the Pseudomonas fluorescens SBW25 wrinkly spreader requires an acetylated form of cellulose. Mol Microbiol 2003; 50:15–27 [View Article]
    [Google Scholar]
  23. Wu C, Lim JY, Fuller GG, Cegelski L. Quantitative analysis of amyloid-integrated biofilms formed by uropathogenic Escherichia coli at the air-liquid interface. Biophys J 2012; 103:464–471 [View Article]
    [Google Scholar]
  24. Rühs PA, Böni L, Fuller GG, Inglis RF, Fischer P. In-situ quantification of the interfacial rheological response of bacterial biofilms to environmental stimuli. PLoS One 2013; 8:e78524 [View Article]
    [Google Scholar]
  25. Hollenbeck EC, Fong JCN, Lim JY, Yildiz FH, Fuller GG et al. Molecular determinants of mechanical properties of V. cholerae biofilms at the air-liquid interface. Biophys J 2014; 107:2245–2252 [View Article]
    [Google Scholar]
  26. Lee KWK, Periasamy S, Mukherjee M, Xie C, Kjelleberg S et al. Biofilm development and enhanced stress resistance of a model, mixed-species community biofilm. ISME J 2014; 8:894 [View Article]
    [Google Scholar]
  27. Burmølle M, Webb JS, Rao D, Hansen LH, Sørensen SJ et al. Enhanced biofilm formation and increased resistance to antimicrobial agents and bacterial invasion are caused by synergistic interactions in multispecies biofilms. Appl Environ Microbiol 2006; 72:3916–3923 [View Article]
    [Google Scholar]
  28. Filoche SK, Anderson SA, Sissons CH. Biofilm growth of Lactobacillus species is promoted by Actinomyces species and Streptococcus mutans. Oral Microbiol Immunol 2004; 19:322–326 [View Article]
    [Google Scholar]
  29. Khare A, Tavazoie S. Multifactorial competition and resistance in a two-species bacterial system. PLoS Genet 2015; 11:e1005715 [View Article]
    [Google Scholar]
  30. Stewart EJ, Payne DE, Ma TM, VanEpps JS, Boles BR et al. Effect of antimicrobial and physical treatments on growth of multispecies Staphylococcal Biofilms. Appl Environ Microbiol 2017; 83:e03483–16 [View Article]
    [Google Scholar]
  31. Ternström A, Lindberg AM, Molin G. Classification of the spoilage flora of raw and pasteurized bovine milk, with special reference to Pseudomonas and Bacillus . J Appl Bacteriol 1993; 75:25–34 [View Article]
    [Google Scholar]
  32. Ribeiro Júnior JC, de Oliveira AM, Silva FdeG, Tamanini R, de Oliveira ALM et al. The main spoilage-related psychrotrophic bacteria in refrigerated raw milk. J Dairy Sci 2018; 101:75–83 [View Article]
    [Google Scholar]
  33. McPhee JDG. Pseudomonas spp. In Roginski H, Fuquay WJ, Fox FP. (editors) Encyclopedia of Dairy Sciences Academic Press; 2002 pp 2340–2350
    [Google Scholar]
  34. Domingue Gauthier V. Inhibition Du Pathogène Des Salmonidés Saprolegnia Parasitica Par Des Bactéries Aquatiques https://papyrus.bib.umontreal.ca 2013
    [Google Scholar]
  35. Ma W, Peng D, Walker SL, Cao B, Gao CH et al. Bacillus subtilis biofilm development in the presence of soil clay minerals and iron oxides. NPJ Biofilms Microbiomes 2017; 3:4 [View Article]
    [Google Scholar]
  36. Branda SS, Chu F, Kearns DB, Losick R, Kolter R. A major protein component of the Bacillus subtilis biofilm matrix. Mol Microbiol 2006; 59:1229–1238 [View Article]
    [Google Scholar]
  37. Erni P, Fischer P, Windhab EJ, Kusnezov V, Stettin H et al. Stress- and strain-controlled measurements of interfacial shear viscosity and viscoelasticity at liquid/liquid and gas/liquid interfaces. Rev Sci Instrum 2003; 74:4916–4924 [View Article]
    [Google Scholar]
  38. Kim GT, Webster G, Wimpenny JWT, Kim BH, Kim HJ et al. Compartmentalization and activity in a microbial fuel cell. J Appl Microbiol 2006; 101:698–710
    [Google Scholar]
  39. Nguyen-Mau SM, Oh SY, Kern VJ, Missiakas DM, Schneewind O. Secretion genes as determinants of Bacillus anthracis chain length. J Bacteriol 2012; 194:3841–3850 [View Article]
    [Google Scholar]
  40. Hölscher T, Bartels B, Lin YC, Gallegos-Monterrosa R, Price-Whelan A et al. Motility, chemotaxis and aerotaxis contribute to competitiveness during bacterial pellicle biofilm development. J Mol Biol 2015; 427:3695–3708 [View Article]
    [Google Scholar]
/content/journal/micro/10.1099/mic.0.000819
Loading
/content/journal/micro/10.1099/mic.0.000819
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error