Organically modified silica (ORMOSILS) thin films produced by sol-gel method were imprinted with two bacterial strains as whole cells in order to develop an easy, fast and specific probe to detect and specifically identify these micro-organisms when present in water samples. An important feature of the imprinting process was the molecular finger-prints left by these microorganisms alongside morphology, into imprinted film cavities. The films also showed high selectivity toward the imprinted template and were able to discriminate between two very close bacterial species ( E. coli and S. typhimurium ). In addition, several central physical parameters of the experimental water solution were examined ( i.e. , pH, ionic strength and the organic load exemplified by NaCl and TOC concentration, respectively). The method sensitivity to different bacterial concentrations was studied by confocal microscopy (CLSM) and quartz crystal microbalance (QCM) tools. Results showed that increased bacterial concentrations favor rapid adsorption onto imprinted sol-gel films with high affinity, while low pH, increased organic load and high ionic concentrations ( i.e. , seawater) interfere with bacteria re-adsorption, reducing detection capability. Under average drinking water chemical composition the method proved to be highly efficient.
C. J. Woodall, “Waterborne Disease-What Are the Primary Killers?” Desalination, Vol. 248, No. 1-3, 2009, pp. 616-621. doi:10.1016/j.desal.2008.05.110
American Public Health Association, American Water Works Association and Water Pollution Control Federation, “Standard Methods for the Examination of Water and Wastewater,” 21st Edition, American Public Health Association, Washington, 2005.
J. V. C. Amstalden, N. R. Cantusio, da S. Neusely and T. J. Hirata, “Comparison of Methods for the Enumeration of Clostridium Perfringens Perfringens Spores in Water,” Water Science and Technology, Vol. 65, No. 2, 2012, pp. 227-232. doi:10.2166/wst.2012.758
W. J. Dore, K. Henshilwood and D. N. Lees, “Evaluation of F-Specific RNA Bacteriophage as a Candidate Human Enteric Virus Indicator for Bivalve Molluscan Shellfish,” Applied and Environmental Microbiology, Vol. 66, No. 4, 2000, pp. 1280-1285. doi:10.1128/AEM.66.4.1280-1285.2000
Y. Kott and E. F. Glona, “Correlating Coliform Bacteria With Escherichia Coli Bacteriophages in Shellfish,” Water & Sewage Works, Vol. 112, No. 11, 1965, pp. 424- 426.
J. M. Pisciotta, D. F. Rath, P. A. Stanek, D. M. Flanery and V. J. Harwood, “Marine Bacteria Cause False-Positive Results in the Colilert-18 Rapid Identification Test for Escherichia Coli in Florida Waters,” Applied and Environmental Microbiology, Vol. 68, No. 2, 2002, pp. 539-544. doi:10.1128/AEM.68.2.539-544.2002
T. Cohen, J. Starosvetsky, U. Cheruti and R. Armon, “Whole Cell Imprinting in Sol-Gel Thin Films for Bacterial Recognition in Liquids: Macromolecular Fingerprinting,” International Journal of Molecular Sciences, Vol. 11, No. 4, 2010, pp. 1236-1252. doi:10.3390/ijms11041236
K. Mosbach, “Molecular Imprinting,” Trends in Biochemical Sciences, Vol.19, No. 1, 1994, pp. 9-14. doi:10.1016/0968-0004(94)90166-X
K. Mosbach and O. Ramstrom, “The Emerging Technique of Molecular Imprinting and Its Future Impact on Biotechnology,” Nature Biotechnology, Vol. 14, No. 2, 1996, pp. 163-170. doi:10.1038/nbt0296-163
G. Carturan, R. Campostrini, S. Diré, V. Scardi and E. De Alteriis “Inorganic Gels for Immobilization of Biocatalysts: Inclusion of Invertase-Active Whole Cells of Yeast (Saccharomyces Cerevisiae) into Thin Layers of SiO2 Gel Deposited on Glass Sheets,” Journal of Molecular Catalysis, Vol. 57, No. 1, 1989, pp. L13-L16. doi:10.1016/0304-5102(89)80121-X
J. Y. Barreau, J. M. Da Costa, I. Desportes, J. Livage, L. Monjour and M. Gentilini, “Fixation and Immunological Reactivity of Parasitic Protozoa in Sol-Gel Matrices,” Comptes Rendus de l'Académie des Science-Series III-Sciences de la Vie, Vol. 317, No. 7, 1994, pp. 653-657.
i>
Sphaerotilus natans<
/i>
CLSM
J. Livage, C. Roux, J. M. Da Costa, I. Desportes and J.F. Quinson, “Immunoassays in Sol-Gel Matrices,” Journal of Sol-Gel Science and Technology, Vol. 7, No. 1-2, 1996, pp. 45-51. doi:10.1007/BF00401882
J. U. Chia, F. Tamanoi and B. J. I. Z. Dunn, “Patterned Hexagonal Arrays of Living Cells in Sol-Gel Silica Films,” Journal of the American Chemical Society, Vol. 122, No. 27, 2000, pp. 6488-6489. doi:10.1021/ja0011515
F. L. Dickert and G. Hayden, “Bioimprinting of Polymers and Sol-Gel Phases. Selective Detection of Yeasts with Imprinted Polymers.” Analytical Chemistry, Vol. 74, No. 6, 2002, pp. 1302-1306. doi:10.1021/ac010642k
F. L. Dickert, P. Lieberzeit and O. Hayden, “Sensor Strategies for Microorganism Detection—From Physical Principles to Imprinting Procedures,” Analytical and Bio- analytical Chemistry, Vol. 377, No. 3, 2003, pp. 540-549. doi:10.1007/s00216-003-2060-5
O. Hayden, C. Haderspock and F. L. Dickert, “Microorganism Detection with Selective Bioimprinted Polymers and Sol-Gels,” International Journal of Bio-Chromatography, Vol. 6, No. 4, 2001, pp. 303-311.
S. G. Andrews and F. J. Millero, “Electrolyte Effects on Attachment of an Estuarine Bacterium,” Applied and En- vironmental Microbiology, Vol. 47, No. 3, 1984, pp. 495-499.
R. Armon, M. Arella and P. Payment, “A Highly Efficient Second-Step Concentration Technique for Bacteriophages and Enteric Viruses Using Ammonium Sulfate and Tween 80,” Canadian Journal of Microbiology, Vol. 34, No. 5, 1988, pp. 651-655.
D. Avnir, “Organic Chemistry within Ceramic Materials: Doped Sol-Gel Materials,” Accounts of Chemical Research, Vol. 28, No. 8, 1995, pp. 328-334. doi:10.1021/ar00056a002
R. Zusman, C. Rotman, M. Ottolenghi and D. Avnir, “Doped Sol-Gel Glasses as Chemical Sensors,” Journal of Non-Crystalline Solids, Vol. 122, No. 1, 1990, pp. 107-109. doi:10.1016/0022-3093(90)90232-B
G. Bitton and K. C. Marshall, “Adsorption of Microorganisms to Surfaces,” Wiley-Interscience, Weinheim, 1980.
C. E. Zobell, “The Effect of Solid Surfaces upon Bacterial Activity,” Journal of Bacteriology, Vol. 46, No. 1, 1943, pp. 39-56.
W. H. Stark, J. Stadler and E. McCoy, “Some Factors Affecting the Bacterial Population of Freshwater Lakes,” Journal of Bacteriology, Vol. 36, No. 1, 1938, pp. 653-654.
H. Heukelekian and A. Heller, “Relation between Food Concentration and Surface for Bacterial Growth,” Journal of Bacteriology, Vol. 40, No. 4, 1940, pp. 547-558.
M. Rosenberg, E. A. Bayer, J. Delarea and E. Rosenberg, “Role of Thin Fimbriae in Adherence and Growth of Acinetobacter Calcoaceticus RAG-1 on Hexadecane,” Applied and Environmental Microbiology, Vol. 44, No. 4, 1982, pp. 929-937.
R. L. Weiss, “Attachment of Bacteria to Sulphur in Extreme Environments,” Journal of General Microbiology, Vol. 77, No. 2, 1973, pp. 501-507.
M. Fletcher and G. D. Floodgate, “An Electron-Micro-scope Demonstration of an Acidic Polysaccharide Involved in the Adhesion of a Marine Bacterium to Solid Surfaces,” Journal of General Microbiology, Vol. 74, No. 2, 1973, pp. 325-334.
G. G. Geesey, W. T. Richardson, H. G. Yeomans, R. T. Irvin and J. W. Costerton, “Microscopic Examination of Natural Sessile Bacterial Populations from an Alpine Stream,” Canadian Journal of Microbiology, Vol. 23, No. 2, 1977, pp. 1733-1736.
W. A. Corpe, “Attachment of Marine Bacteria to Solid Surfaces,” In: R. S. Manley, Ed., Adhesion in Biological Systems, Academic Press, New York, 1970, pp. 73-87.
M. Fletcher, “The Attachment of Bacteria to Surfaces in Aqueous Environments,” In: D. C. Ellwood, J. Melling and P. Rutter, Eds., Adhesion of Microorganisms to Surfaces, Academic Press, London, 1979, pp. 88-108.
C. M. Brown, D. C. Ellwood and J. R. Hunter, “Growth of Bacteria at Surfaces: Influence of Nutrient Limitation,” FEMS Microbiology Letters, Vol. 1, No. 3, 1977, pp. 163-166.
I. T. Miettinen, T. Vartiainen and P. J. Martikainen, “Phosphorus and Bacterial Growth in Drinking Water,” Applied and Environmental Microbiology, Vol. 63, No. 8, 1997, pp. 3242-3245.
S. G. Andrews and F. J. Millero, “Electrolyte Effects on Attachment of an Estuarine Bacterium,” Applied and Environmental Microbiology, Vol. 47, No. 3, 1984, pp. 495-499.
T. Kaneko and R. R. Colwell, “Adsorption of Vibrio Parahaemolyticus onto Chitin and Copepods,” Applied microbiology, Vol. 29, No. 2, 1975, pp. 269-274.
S. L. Daniels, “The Adsorption of Microorganisms onto Solid Surfaces: A Review,” Developments in Industrial Microbiology Series, Vol. 13, 1972, pp. 211-253.
J. Wood, “Interaction of Micro-Organisms with Ion Exchange Resins,” In: R. C. W. Berkeley, J. M. Lynch, J. Melling, P. R. Rutter and B. Vincent, Eds., Microbial Adhesion to Surfaces, Ellis-Horwood, West Sussex, 1980, pp 163-186.
T. Takeuchi, T. Mukawa and H. Asanuma, “Molecular Imprinting from Fundamentals to Applications,” Wiley-VCH, Weinheim, 2003.