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Deletion of EFH1 Alters Murine Intestinal Colonization

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posted on 2013-02-21, 11:25 authored by Sarah Jane White, Ari Rosenbach, Paul Lephart, Diem Nguyen, Alana Benjamin, Saul Tzipori, Malcolm Whiteway, Joan Mecsas, Carol A Kumamoto

WT, efh1 deletion mutant, or EFH1 reconstituted null mutant were orally inoculated by gavage into Swiss Webster mice. At various days post-inoculation, the amounts of C. albicans in fecal pellets and in organs of the intestinal tract were measured.

(A) CFUs per gram of cecum contents from mice sacrificed on the indicated days post-inoculation. Each symbol represents a sample from a different mouse. (Composite results from several experiments.) Red circles, WT C. albicans strain DAY185 (23 mice); blue triangles, efh1 deletion mutant strain CKY366 (24 mice); black diamonds, EFH1 reconstituted strain, CKY373 (ten mice); open symbols, no colonies detected; bars, geometric means. p-Value was determined using the t test with log transformed data. ** indicates p < 0.000005.

(B) CFUs per gram of fecal pellet. Mice were sampled repeatedly and each symbol represents a sample from a different mouse. Different numbers of mice are shown on different days because some mice were sacrificed earlier and some mice were sampled on different days. Red circles, WT C. albicans strain DAY185 (ten mice); blue triangles, efh1 deletion mutant strain CKY366 (12 mice); black diamonds, EFH1 reconstituted strain, CKY373 (seven mice); open symbols, no colonies detected; bars, geometric means. p-Value was determined using the t test with log transformed data. * indicates p < 0.0003.

(C) Correlation between CFUs measured in several organs and in fecal pellets. Light blue diamonds, fecal pellets (labeled F); pink triangles, cecum contents (labeled C); open circles, ileum contents (labeled I); squares, stomach contents (labeled S). CFU/gm of cecum contents, stomach contents, and fecal pellets correlated well, while the CFU/gm ileum was more variable.

(D) Mice were colonized with WT strain DAY185 (red circles) or efh1 null mutant CKY366 (blue triangles) or were not inoculated with C. albicans (not shown). Mice were sacrificed on day 21 post-inoculation, CFUs from cecum contents were determined by plating, and C. albicans DNA in cecum contents was quantified by qPCR as described in Materials and Methods. The correlation between CFUs in 70 mg of pelleted cecum contents (x-axis) and DNA (in arbitrary units) determined by qPCR (y-axis) is shown. Each symbol represents a different mouse.

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