In Vitro Aging of Helicobacter pylori: Changes in Morphology, Intracellular Composition and Surface Properties

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Abstract
Background. During the conversion from the bacillary into the coccoid form, Helicobacter pylori organisms are known to change extensively. The aim of this study was to determine some of the changes that occur regarding morphology, intracellular composition and surface properties during the aging of bacteria in vitro. Materials and Methods. H. pylori from agar plate cultures of different ages was used in this study. The intracellular composition of the two morphological forms of the bacteria was tested by density centrifugation, DNA extraction and quantitative OD, mRNA and ATP measurements. Immunoblotting was used to observe changes in secreted/superficial protein patterns, and hydrophobicity measurements were used to observe changes in surface properties. Results. All bacillary H. pylori organisms changed morphology gradually over 10 days of culture. Rods had a higher density than cocci; bacteria stored in PBS had the highest density and bacteria stored in water had the lowest. The quantitative DNA, RNA and ATP content were reduced in the aging bacteria. Fewer immunogenic proteins were expressed, and an increased surface hydrophobicity was observed in the older cultures. Conclusion. This study highlights several aspects of H. pylori aging in vitro and shows some of the differences that exist between bacillary and coccoid forms. This information is important for understanding the transmission and survival of H. pylori outside the human host, as the degradative changes in the intracellular composition and the surface properties shown here point to dead bacteria, and not to a viable but nonculturable form.