Methods, Models, and Analysis of Bacterial Adhesion

  • Itzhak Ofek
  • Ronald J. Doyle


It is axiomatic to consider that most living and nonliving surfaces have a tendency to be colonized by microorganisms. The importance of microbial adhesion and colonization to surfaces was not appreciated until molecular techniques were applied to analyze modes and mechanisms of cell—substratum interactions. As more and more techniques became available, new knowledge was gained that made it possible to understand the modulation of the adhesion and subsequent colonization of many microorganisms. To date, no single experimental system has been developed that can be used to adequately characterize all aspects of microbe—substratum interactions. It is therefore essential that the reliabilities, advantages, and limitations of the existing techniques be understood. Most techniques employed in the study of adhesion yield restricted amounts of information, usually about defined events in a complicated series of interactions. This chapter considers methods for the study of adhesion. Consideration is given to model systems, methods for separating adherent from nonadherent cells, controlled and uncontrolled variables in experimental design, and approaches used in analyzing adhesion data. Finally, methods related to the identification and regulation of expression of adhesins and their receptors are reviewed.


Bacterial Adhesion Tissue Culture Cell Adherent Bacterium Positive Cooperativity Adhesion Experiment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Albertsson, P.A. 1958. Particle fractionation in liquid two-phase system. The composition of some phase systems and the behavior of some model particles in them. Application to the isolation of cell walls from microorganisms. Biochim. Biophys. Acta 27: 378–395.PubMedCrossRefGoogle Scholar
  2. Allison, D.G. and I.W. Sutherland. 1984. A staining technique for attached bacteria and its correlation to extracellular carbohydrate production. J. Microbiol. Meth. 2: 93–99.CrossRefGoogle Scholar
  3. Aronson, M., O. Medalia, L. Schon, D. Mirelman, N. Sharon, and I Ofek. 1979. Prevention of colonization of the urinary tract of mice with Escherichia coliby blocking of bacterial adherence with methyl alpha-D-mannopyranoside. J. Infect. Dis. 139: 329–332.PubMedCrossRefGoogle Scholar
  4. Athamna, A. and I Ofek. 1988. Enzyme-linked immunosorbent assay for quantitation of attachment and ingestion stages of bacterial phagocytosis. J. Clin. Microbiol. 26: 62–66.PubMedGoogle Scholar
  5. Baselsky, V.S. and C.D. Parker. 1978. Intestinal distribution of Vibrio cholerae in orally infected infant mice: kinetics of recovery of radiolabel and viable cells. Infect. Immun. 21: 518–525.Google Scholar
  6. Beachey, E.H. and I. Ofek. 1976. Epithelial cell binding of group A streptococci by lipoteichoic acid on fimbriae denuded of M proteins. J. Exp. Med. 143: 759–771.PubMedCrossRefGoogle Scholar
  7. Brennan, M.J., J.H. Hannah and E. Leininger. 1991. Adhesion of Bordetella pertussis to sulfatides and to the GalNacß4Gal sequence found in glycosphingolipids. J. Biol. Chem. 266: 18827–18831.PubMedGoogle Scholar
  8. Clark, W.B., L.L. Bammann, and R.J. Gibbons. 1978. Comparative estimates of bacterial affinities and adsorption sites on hydroxyapatite surfaces. Infect. Immun. 19: 846–853.PubMedGoogle Scholar
  9. Costerton, J.W. 1980. Some techniques involved in study of adsorption of microorganisms to surfaces. In: Bitton, G. and K.C. Marshall (eds.), Adsorption of Microorganisms to Surfaces. John Wiley & Sons, New York, pp. 403–423.Google Scholar
  10. Cowan, M.M., K.G. Taylor, and R.J. Doyle. 1986. Kinetic analysis of Streptococcus sanguis adhesion to artificial pellicle. J. Dent. Res. 65: 1278–1283.PubMedCrossRefGoogle Scholar
  11. Cowan, M.M., K.G. Taylor, and R.J. Doyle. 1987a. Role of sialic acid in the kinetics of Streptococcus sanguis adhesion to artificial pellicle. Infect. Immun. 55: 1552–1557.Google Scholar
  12. Cowan, M.M., K.G. Taylor, and R.J. Doyle. 1987b. Energetics of the initial phase of adhesion of Streptococcus sanguisto hydroxylapatite. J. Bacteriol. 169: 2995–3000.Google Scholar
  13. de Man, P., B. Cedergren, S. Enerback, A.C. Larsson, H. Leffler, A.L. Lundell, B. Nilsson, and C. Svanborg-Eden. 1987. Receptor-specific agglutination tests for detection of bacteria that bind globoseries glycolipids. J. Clin. Microbiol. 25: 401–406.PubMedGoogle Scholar
  14. Doyle, R.J. 1991. Strategies in experimental microbial adhesion research. In: Mozes, N, P.S. Handley, H.J. Busscher, and P.G. Rouxhet (eds.), Microbial Cell Surface analysis-Structural and Physiocochemical Methods. VCH Publishers, New York, pp. 293–316.Google Scholar
  15. Doyle, R.J., J.D. Oakley, K.R. Murphy, D. McAlister, and K.G. Taylor. 1985. Graphical analyses of adherence data. In: Mergenhagen, S.E. and B. Rosan (eds.), Molecular Basis of Oral Microbial Adhesion. American Society for Microbiology, Washington, pp. 109–113.Google Scholar
  16. Drake, D., K.G. Taylor, A.S. Bleiweis, and R.J. Doyle. 1988. Specificity of the glucanbinding lectin of Streptococcus cricetus. Infect. Immun. 56: 1864–1872.Google Scholar
  17. Duguid, J.P. and R.R. Gillies. 1957. Fimbriae and adhesive properties of dysentery bacilli. J. Pathol. Bacteriol. 74: 397–411.CrossRefGoogle Scholar
  18. Duguid, J.P. and D.C. Old. 1980. Adhesive properties of Enterobacteriaceae. In: Beachey, E.H. (ed.), Bacterial Adherence (Receptors and Recognition, Vol. 6). London, Chapman and Hall, pp. 184–217.Google Scholar
  19. Duguid, J.P., M.R. Darekar, and D.W.F. Wheater. 1976. Fimbriae and infectivity in Salmonella typhimurium. J. Med. 11ficrobiol. 9: 459–473.Google Scholar
  20. Elbein, A.D., B.A. Sanford, M.A. Ramsey, and Y.T. Pan. 1981. Effect of inhibitors of glycoprotein biosynthesis and bacterial adhesion. In: Elliot, K., M. O’Connor, and J. Whelan (eds.), Adhesion and Microorganism Pathogenicity (Ciba Foundation Symposium 80). Pitman Medical, London, pp. 270–282.Google Scholar
  21. Ellen, R.P. and R.J. Gibbons. 1972. M protein-associated adherence of Streptococcus pyogenesto epithelial surfaces: prerequisite for virulence. Infect. Immun. 5: 826–830.PubMedGoogle Scholar
  22. Ellen, R.P. and R.J. Gibbons. 1973. Parameters affecting the adherence and tissue tropisms of Streptococcus pyogenes. Infect. Immun. 9: 85–91.Google Scholar
  23. Filler, S.G., L.G. Der, C.L. Mayer, P.D. Christenson, and J.E. Edwards, Jr. 1987. An enzyme-linked immunosorbent assay for quantifying adherence of Candidato human vascular endothelium. J. Infect. Dis. 156: 561–566.PubMedCrossRefGoogle Scholar
  24. Firon, N., D. Duksin, and N. Sharon. 1985. Mannose-specific adherence of Escherichia colito BHK cells that differ in their glycosylation patterns. FEMS Microbiol. Lett. 27: 161–165.CrossRefGoogle Scholar
  25. Goldhar, J., A. Zilberberg, and I. Ofek. 1986. Infant mouse model of adherence and colonization of intestinal tissues by enterotoxigenic strains of Escherichia coliisolated from humans. Infect. Immun. 52: 205–208.PubMedGoogle Scholar
  26. Hjerten, S., J. Rosengren, and S. Pahlman. 1974. Hydrophobic interaction chromatography. The synthesis and the use of some alkyl and aryl derivatives of agarose. J. Chromatogr. 101: 281–288.CrossRefGoogle Scholar
  27. Karlsson, J. 1986. Animal glycolipids as attachment sites for microbes. Chem. Phys. Lipids 42: 153–172.PubMedCrossRefGoogle Scholar
  28. Klotz, I.M. 1982. Numbers of receptor sites from Scatchard graphs: facts and fantasies. Science 217: 1247–1249.PubMedCrossRefGoogle Scholar
  29. Kluepfel, D.A. and S.G. Pueppke. 1985. Isotherm for adsorption of Agrobacterium tumefaciens to susceptible potato (Solanum tuberosum L.) tissues. Appl. Environ. Microbiol. 49: 1351–1355.PubMedGoogle Scholar
  30. Liljemark, W.F., C.G. Bloomquist, and L.J. Fenver. 1985. Characteristics of the adherence of oral Hemophilius species to an experimental salivary pellicle and to other oral bacteria, In: Mergenhagen, S. and B. Rosan (eds.), Molecular Basis of Oral Microbial Adhesion. American Society for Microbiology, Washington, pp. 94–102.Google Scholar
  31. Ludwicka, A., L.M. Switalski, A. Lundin, G. Pulverer, and T. Wadstrom. 1985. Bioluminescent assay for measurement of bacterial attachment to polyethylene. J. Microbiol. Methods 4: 169–177.CrossRefGoogle Scholar
  32. Mackowiak, P.A. and M. Marling-Cason. 1984. A comparative analysis of in vitro assays of bacterial adherence. J. Microbiol. Methods 2: 147–158.CrossRefGoogle Scholar
  33. McEachran, D.W. and R.T. Irvin. 1986. A new method for the irreversible attachment of cells or proteins to polystyrene tissue culture plates for use in the study of bacterial adhesion. J. Microbiol. Methods 5: 99–111.CrossRefGoogle Scholar
  34. Myerthall, D.L. and T.H. Thomas. 1983. Kinetics of adherence of Actinomyces viscosus to saliva-coated silica and hydroxyapatite beads. J. Gen. Microbiol. 129: 1387–1395.PubMedGoogle Scholar
  35. Nesbitt, W.E., R.J. Doyle, K.G. Taylor, R.H. Staat, and R.R. Arnold. 1982. Positive cooperativity in the binding of Streptococcus sanguis to hydroxylapatite. Infect. Immun. 35: 157–165.PubMedGoogle Scholar
  36. Ofek, I. and E.H. Beachey. 1978. Mannose binding and epithelial cell adherence of Escherichia coli. Infect. Immun. 22: 247–254.PubMedGoogle Scholar
  37. Ofek, I., D. Mirelman, and N. Sharon. 1977. Adherence of Escherichia colito human mucosal cells mediated by mannose receptors. Nature 265: 923–625.CrossRefGoogle Scholar
  38. Ofek, I., H.S. Courtney, D.M. Schifferli, and E.H. Beachey. 1986. Enzyme-linked immunosorbent assay for adherence of bacteria to animal cells. J. Clin. Microbiol. 24: 512–516.PubMedGoogle Scholar
  39. Pooniah, S., S.N. Abraham, M.E. Dokter, C.D. Wall, and R.D. Endres. 1989. Mitogenic stimulation of human lymphocytes by the mannose-specific adhesin on Escherichia coli type 1 fimbriae. J. Immunol. 142: 992–998.Google Scholar
  40. Reid, G. 1989. Local and diffuse bacterial adherence on uropithelial cells. Curr. Microbiol. 18: 93–97.CrossRefGoogle Scholar
  41. Rosan, B., R. Eifert, and E. Golub. 1985. Bacterial surfaces, salivary pellicles and plaque formation. In: Mergenhagen, S.E. and B. Rosan (eds.), Molecular Basis of Oral Microbial Adhesion. American Society for Microbiology, Washington, pp. 69–76.Google Scholar
  42. Rosenberg, M. 1981. Bacterial adherence to polystyrene: a replica method of screening for bacterial hydrophobicity. Appl. Environ. Microbiol. 42: 375–377.PubMedGoogle Scholar
  43. Rosenberg, M., D. Gutnick, and E. Rosenberg. 1980. Adherence of bacteria to hydrocarbons: a simple method for measuring cell surface hydrophobicity. FEMS Microbiol. Leu. 9: 29–33.CrossRefGoogle Scholar
  44. Rosenberg, M.A., E.A. Perry, D.L. Gutnick, E. Rosenberg, and I. Ofek. 1981. Adherence of Acinetobacter calcoacetius RAG-1 to human epithelial cells and to hexadecane. Infect. Immun. 33: 29–33.PubMedGoogle Scholar
  45. Rosenstein, I.J., D. Grady, J.M.T. Hamilton-Miller, and W. Brumfitt. 1985. Relationship between adhesion of Escherichia colito uro-epithelial cells and the pathogenesis of urinary infection: problems in methodology and analysis. J. Med. Microbiol. 20: 335–344.PubMedCrossRefGoogle Scholar
  46. Scaletsky, C.A., M. Silva, and R. Trabulsi, 1984. Distinctive patterns of adherence of enteropathogenic Escherichia colito HeLa cells. Infect. Immun. 45: 534–536.PubMedGoogle Scholar
  47. Schadow, K.H., W.A. Simpson, and S.D. Christensen. 1988. Characteristics of adherence to plastic tissue culture plates of coagulase negative staphylococci exposed to subinhibitory concentrations of antibiotics. J. Infect. Dis. 157: 71–77.PubMedCrossRefGoogle Scholar
  48. Smyth, C.J., P. Jonsson, E. Olsson, O. Soderlind, R. Rosengren, S. Hjerten, and T. Wadstrom. 1978. Differences in hydrophobic surface characteristics of porcine enteropathogenic Escherichia coliK88 antigen as revealed by hydrophobic interaction chromatography. Infect. Immun. 22: 462–472.PubMedGoogle Scholar
  49. Stanislawski, L., W.A. Simpson, D.L. Hasty, N. Sharon, E.H. Beachey, and I. Ofek. 1985. Role of fibronectin in attachment of Streptococcus pyogenes and Escherichia coli to human cell lines and isolated oral epithelial cells. Infect. Immun. 48: 257–259.PubMedGoogle Scholar
  50. Svanborg-Eden, C. R. Freter, L. Hagberg, R. Hull, S. Hull, H. Leffler, and G. Schoolnik. 1982. Inhibition of experimental ascending urinary tract infection by an epithelial cell-surface receptor analogue. Nature 298: 560–562.CrossRefGoogle Scholar
  51. Valentin-Weigand, P., G.S. Chhatwal, and H. Blobel. 1987. A simple method for quantitative determination of bacterial adherence to human and animal epithelial cells. Microbiol. Immunol. 31: 1017–1023.Google Scholar
  52. Woolfson, A.D., S.P. Gorman, D.F. McCafferty, and D.S. Jones. 1987. On the statistical evaluation of adherence assays. J. Appl. Bacteriol. 63: 147–151.PubMedCrossRefGoogle Scholar
  53. Wyatt, J.E., S.M. Poston, and W.C. Noble. 1990. Adherence of Staphylococcus aureus to cell monolayers. J. Appl. Bacteriol. 69: 834–844.PubMedCrossRefGoogle Scholar
  54. Zilberberg, A., J. Goldhar, and I. Ofek. 1983. Adherence of enterotoxigenic Escherichia coli (ETEC) strains to mouse intestine segments analyzed by Langmuir adherence isotherms. FEMS Microbiol. Lett. 16: 225–228.CrossRefGoogle Scholar
  55. Zilberberg, A., I. Ofek, and J. Goldhar. 1984. Affinity of adherence in vitro and colonization of mice intestine by enterotoxigenic Escherichia coli (ETEC). FEMS Microbiol. Lett. 23: 103–106.CrossRefGoogle Scholar

Copyright information

© Chapman & Hall, Inc. 1994

Authors and Affiliations

  • Itzhak Ofek
    • 1
  • Ronald J. Doyle
    • 2
  1. 1.Tel-AvivIsrael
  2. 2.LouisvilleUSA

Personalised recommendations