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Research Journal of Microbiology
Year: 2015  |  Volume: 10  |  Issue: 4  |  Page No.: 158 - 169

EDTA-Treated Cell Membrane and Multiple Subculturing Affect the Virulence of Enteropathogenic Bacteria

O.S. Folorunso, K.O. Amisu and B.F. Ogungbe    

Abstract: The ability of microbial cell membrane to bind organic dye is an age long protocol that preludes the identification and characterization of bacteria cells and its components. We employed formation of keratoconjunctivitis, pigmentations and Congo red binding assay to assess the virulence of eight enteric pathogenic bacteria. Staphylococcus aureus and Salmonella typhimurium, at 3.0×108 CFU mL-1, was sufficient to cause keratoconjunctivitis in the eyes of Albino rat (Sereny test), while observing traces in Streptococcus faecalis, Klebsiella pneumoniae, Pseudomonas aeruginosa and Bacillus subtilis. Pigmentation (PCr+) occurred in Staphylococcus aureus, Streptococcus faecalis and Klebsiella pneumoniae. Highest protein secretion was found in Escherichia coli. Hydrophobicity of the microbial cell membrane increased in the order of chemical pre-treatments (0.9% w/v physiological saline, 50 mM Tris HCl, pH 7.8, 50 mM Tris HCl, pH 7.8+50 mM EDTA). Thus, Tris-EDTA adjunct increased hydrophobicity of the membrane to bind more Congo red than other treatments. Virulence strains of Gram-positive bacteria may likely cause a more severe keratoconjunctivitis. Pigmentations favoured Gram-positive than negative probably because of the membrane differentiation. The effect of multiple subculturing on the Congo red binding ability of these bacteria remained inconclusive as less than half of the tested bacteria complied with our hypothesis though, all were expectedly affected, when pre-treated. The virulence of bacteria is pre-determined by the inherent plasmid factors but their expression is a function of the bacteria cells to bind organic dye. Therefore, physico-chemical treatments that promote hydrophobicity of the bacteria cell membrane are likely to reduce their virulence.

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