A comparative study of biofilm formation by Shiga toxigenic Escherichia coli using epifluorescence microscopy on stainless steel and a microtitre plate method

Rivas, Lucia, Dykes, Gary A. and Fegan, Narelle (2007) A comparative study of biofilm formation by Shiga toxigenic Escherichia coli using epifluorescence microscopy on stainless steel and a microtitre plate method. Journal of Microbiological Methods, 69 1: 44-51.


Author Rivas, Lucia
Dykes, Gary A.
Fegan, Narelle
Title A comparative study of biofilm formation by Shiga toxigenic Escherichia coli using epifluorescence microscopy on stainless steel and a microtitre plate method
Formatted title A comparative study of biofilm formation by Shiga toxigenic Escherichia coli using epifluorescence microscopy on stainless steel and a microtitre plate method
Journal name Journal of Microbiological Methods  (ERA 2012 Listed)    (ERA 2010 Rank B)   Check publisher's open access policy
Publication date 2007-04
Sub-type Article
DOI 10.1016/j.mimet.2006.11.014
Volume number 69
Issue number 1
ISSN 0167-7012; 1872-8359
Start page 44
End page 51
Total pages 8
Editor A. Fox
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Language eng
Formatted abstract Attachment of Shiga toxigenic Escherichia coli (STEC) to surfaces and the formation of biofilms may enhance persistence in a food processing environment and present a risk of contaminating products. Seven strains of STEC and three non-STEC strains were selected to compare two biofilm quantification methods; epifluorescence microscopy on stainless steel (SS) and a microtitre plate assay. The influence of prior growth in planktonic (nutrient broth) and sessile (nutrient agar) culture on biofilm production, as well as expression of surface structures and the possession of antigen 43 (encoded by agn43) on biofilm formation were also investigated. Biofilms were produced in diluted nutrient broth at 25 °C for 24 and 48 h. Curli expression was determined using congo red indicator agar, while the presence of agn43 was determined using polymerase chain reaction. No correlation was found between counts for epifluorescence microscopy on SS and the absorbance values obtained with the microtitre plate method for planktonic and sessile grown cultures. Different abilities of individual STEC strains to attach to SS and microtitre plates were found with some strains attaching better to each surface following growth in either planktonic or sessile culture. All O157 STEC strains had low biofilm counts on SS for planktonic and sessile grown cultures; however, one STEC O157:H− strain (EC516) had significantly greater (p < 0.05) biofilm production on microtitre plates compared to the other O157 STEC strains. EC516 and other STEC (O174:H21 and O91:H21) strains expressing curli fimbriae were found to produce significantly greater (p < 0.05) biofilms on microtitre plates compared to the non-curli expressing strains. No relationship was found between the production of type-I fimbriae, motility, agn43 and bacterial physicochemical properties (previously determined) and biofilm formation on SS or microtitre plates. Variations between the two biofilm determination methods may suggest that the biofilm production on microtitre plates may not be appropriate to represent other surfaces such as SS and that caution should be taken when selecting a method to quantify biofilm production on a surface.
Keyword Biochemical research methods
Microbiology
Biofilm formation
Epifluorescence microscopy
Escherichia coli
Microtitre plate
Stainless steel
Listeria-monocytogenes strains
Physicochemical properties
Abiotic surfaces
Curli production
Attachment
Adhesion
O157:H7
Food
Adherence
Fimbriae
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article
Collection: School of Agriculture and Food Sciences
 
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Created: Mon, 18 Feb 2008, 16:55:24 EST