Advertisement

Antonie van Leeuwenhoek

, Volume 103, Issue 1, pp 251–259 | Cite as

Air–liquid interface biofilm formation by psychrotrophic pseudomonads recovered from spoilt meat

  • Mhari Robertson
  • Simona M. Hapca
  • Olena Moshynets
  • Andrew J. Spiers
Short Communication

Abstract

The ability to colonise the surface of liquids has obvious advantages for bacteria and biofilm formation at the meniscus and air–liquid (A–L) interface is common amongst environmental pseudomonads. Bacteria from this genus also colonise raw meat and in this work the ability of these to produce biofilms was assessed. Sixty isolates were recovered from vacuum-packed venison, phenotypically characterised and shown by hierarchical cluster analysis to represent a diverse collection of psychrotrophic spoilt venison-associated pseudomonads. Of these, 12 % were found to produce biofilms limited to the meniscus region of the microcosm walls, 31 % produced attached biofilms with significant extensions across the A–L interface and 45 % produced unattached ‘floating’ biofilms. A combined statistical analysis of growth, biofilm strength and attachment levels revealed that growth affected strength but not attachment, and that there was a significant relationship between attachment and strength. Some environmental pseudomonads are known to utilise cellulose as a biofilm matrix component and here 28 % of the SVP isolates were found to express cellulose by epifluorescent microscopy. This survey suggests that biofilm formation may be more common in psychrotrophic meat-associated isolates than amongst the wider pseudomonad community from which spoilage bacteria might be recruited. This may reflect a selective advantage of bacterial aggregations such as biofilms in environments subject to high levels of physical disturbance. Aggregations may be more resistant to competition and dehydration stress than individual bacteria, whilst fragments of these aggregations may prove more effective in the colonisation of new habitats.

Keywords

Biofilm Cellulose Pseudomonas Psychrotrophic Spoilt meat 

Notes

Acknowledgments

We thank Vasiliy Gorchev and Sergey Karakhim of the Laboratory of Optical Methods Investigation at the Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, for their help with the confocal laser scanning microscopy. AS is funded by the University of Abertay Dundee and is a member of the Scottish Alliance for Geoscience, Environment and Society (SAGES). Additional funding from the Royal Society of Edinburgh through the International Exchange Programme helped support the collaboration between AS and OM. The University of Abertay Dundee is a charity registered in Scotland, No: SC016040.

Conflict of interest

None.

Supplementary material

10482_2012_9796_MOESM1_ESM.pdf (43 kb)
Supplementary material 1 (PDF 452 kb)

References

  1. Alhede M, Kragh KN, Qvortrup K, Allesen-Holm M, Van Gennip M, Christensen LD, Jensen PO, Nielsen AK, Parsek M, Wozniak D, Molin S, Tolker-Nielsen T, Høiby N, Givskov M, Bjarnsholt T (2011) Phenotypes of non-attached Pseudomonas aeruginosa aggregates resemble surface attached biofilm. PLoS ONE 6:e27943PubMedCrossRefGoogle Scholar
  2. Bantinaki E, Kassen R, Knight C, Robinson Z, Spiers AJ, Rainey PB (2007) Adaptive divergence in experimental populations of Pseudomonas fluorescens. III. Mutational origins of wrinkly spreader diversity. Genetics 176:441–453PubMedCrossRefGoogle Scholar
  3. Borch E, Kant-Muemansb M-L, Blixt Y (1996) Bacterial spoilage of meat products and cured meat. Int J Food Microbiol 33:103–120PubMedCrossRefGoogle Scholar
  4. Branda SS, Vik A, Friedman Kolter R (2005) Biofilms: the matrix revisited. Trends Microbiol 13:20–26PubMedCrossRefGoogle Scholar
  5. Coenye T, Nelis HJ (2010) In vitro and in vivo model systems to study microbial biofilm formation. J Microbiol Methods 83:89–105PubMedCrossRefGoogle Scholar
  6. Djordjevic D, Wiedmann M, McLandsborough LA (2002) Microtitre plate assay for assessment of Listeria monocytogenes biofilm formation. Appl Environ Microbiol 68:2950–2958PubMedCrossRefGoogle Scholar
  7. Donlan RM, Costerton JW (2002) Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 15:167–193PubMedCrossRefGoogle Scholar
  8. Fox J (2008) Applied regression analysis and generalized linear models, 2nd edn. SAGE, LondonGoogle Scholar
  9. Gehrig SM (2005) Adaptation of Pseudomonas fluorescens SBW25 to the air-liquid interface: a study in evolutionary genetics. University of Oxford, OxfordGoogle Scholar
  10. Gill CO (2007) Microbiological conditions of meats from large game animals and birds. Meat Sci 77:149–160PubMedCrossRefGoogle Scholar
  11. King EO, Ward MK, Raney DC (1954) Two simple media for the demonstration of pyocyanin and fluorescin. J Lab Clin Med 44:301–307PubMedGoogle Scholar
  12. Koza A, Hallett PD, Moon CD, Spiers AJ (2009) Characterisation of a novel air–liquid interface biofilm of Pseudomonas fluorescens SBW25. Microbiology 155:1397–1406PubMedCrossRefGoogle Scholar
  13. Koza A, Moshynets O, Otten W, Spiers AJ (2011) Environmental modification and niche construction: developing O2 gradients drive the evolution of the Wrinkly Spreader. Int Soc Microb Ecol J 5:665–673Google Scholar
  14. Labadie J (1999) Consequences of packaging on bacterial growth. Meat is an ecological niche. Meat Sci 52:299–305PubMedCrossRefGoogle Scholar
  15. Lebert I, Begot C, Lebert A (1998) Growth of Pseudomonas fluorescens and Pseudomonas fragi in a meat medium as affected by pH (5.8–7.0), water activity (0.97–1.00) and temperature (7–25 °C). Int J Food Microbiol 39:53–60PubMedCrossRefGoogle Scholar
  16. McLandsborough L, Rodriguez A, Pérez-Conesa D, Weiss J (2006) Biofilms: at the interface between biophysics and microbiology. Food Biophys 1:94–114CrossRefGoogle Scholar
  17. Molin G, Ternström A (1982) Numerical taxonomy of psychrotrophic pseudomonads. J Gen Microbiol 128:1249–1264PubMedGoogle Scholar
  18. Molin G, Ternström A (1986) Phenotypically based taxonomy of psychrotrophic Pseudomonas isolated from spoiled meat, water, and soil. Int J Syst Bacteriol 36:257–274CrossRefGoogle Scholar
  19. Morita RY (1975) Psychrophilic bacteria. Bacterial Rev 39:144–167Google Scholar
  20. Moshynets OV, Koza A, Dello Sterpaio P, Kordium VA, Spiers AJ (2011) Up-dating the Cholodny method using PET films to sample microbial communities in soil. Biopol Cell 27:199–205Google Scholar
  21. Nychas G-JE, Skandamis PN, Tassou CC, Koutsoumanis KP (2008) Meat spoilage during distribution. Meat Sci 78:77–89PubMedCrossRefGoogle Scholar
  22. Peix A, Ramírez-Bahena M-H, Velázquez E (2009) Historical evolution and current status of the taxonomy of genus Pseudomonas. Infect Genet Evol 9:1132–1147PubMedCrossRefGoogle Scholar
  23. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. CSH Laboratory Press, Cold Spring HarborGoogle Scholar
  24. Solomon EB, Miemira BA, Sapers GM, Annous BA (2005) Biofilm formation, cellulose production, and curli biosynthesis by Salmonella originating from produce, animal, and clinical sources. J Food Prot 68:906–112Google Scholar
  25. Spiers AJ, Kahn SG, Travisano M, Bohannon J, Rainey PB (2002) Adaptive divergence in experimental populations of Pseudomonas fluorescens. I. Genetic and phenotypic bases of Wrinkly Spreader fitness. Genet 161:33–46Google Scholar
  26. Spiers AJ, Bohannon J, Gehrig S, Rainey PB (2003) Biofilm formation at the air–liquid interface by the Pseudomonas fluorescens SBW25 wrinkly spreader requires an acetylated form of cellulose. Mol Microbiol 50:15–27PubMedCrossRefGoogle Scholar
  27. Sutherland IW (2001) Biofilm exopolysaccharides: a strong and sticky framework. Microbiology 147:3–9PubMedGoogle Scholar
  28. Ude S, Arnold DL, Moon CD, Timms-Wilson T, Spiers AJ (2006) Biofilm formation and cellulose expression among diverse environmental Pseudomonas isolates. Environ Microbiol 8:1997–2011PubMedCrossRefGoogle Scholar
  29. Van Houdt R, Michiels CW (2010) Biofilm formation and the food industry, a focus on the bacterial outer surface. J Appl Microbiol 109:1117–1131PubMedCrossRefGoogle Scholar
  30. Verran J (2002) Biofouling in food processing. Biofilm or biotransfer potential? Trans IChemE 80:292–298Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Mhari Robertson
    • 1
  • Simona M. Hapca
    • 1
  • Olena Moshynets
    • 2
  • Andrew J. Spiers
    • 1
  1. 1.The SIMBIOS Centre & School of Contemporary SciencesUniversity of Abertay DundeeDundeeUK
  2. 2.Institute of Molecular Biology and Genetics of the National Academy of Sciences of UkraineKievUkraine

Personalised recommendations