Reduced lipopolysaccharide O antigen expression, increased acid susceptibility and multicellular behaviour in an Escherichia coli isolate after long-term in vitro exposure to formic acid

Organic acids are being used in animal husbandry due to their positive effects on overall health and growth rate in animal husbandry. However, little is known about the long-term effects of organic acids on the normal microbiota. In this work, an Escherichia coli strain isolated from a healthy pig was passed weekly for 20 months in medium with and without formic acid (FA) to investigate the effects of long-term exposure to organic acids. Passages were made using buffered medium at pH 5.25 and 6.1 with and without FA. Colonies showing reduced O antigen lipopolysaccharide production and increased susceptibility to FA were isolated after 12 months exposure to FA at pH 5.25 and after 20 months exposure to FA at pH 6.1. Adaptation to acid and bacterial growth was strongly affected by FA at pH 5.25. Also, after 2 months of serial passages, we observed colonies exhibiting multicellular behaviour, with production of fimbriae and an extracellular matrix. We conclude that long-term exposure to FA of an E. coli isolate from the normal intestinal microbiota is associated with various changes in the bacterial cell surface involving reduced O antigen expression and increased extracellular matrix production together with increased susceptibility to FA.