Severe protein malnutrition in mice selectively enhances intestinal disruption and severity of cryptosporidiosis.
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Abstract
<p>(A) Experimental timeline. 3-week-old C57Bl/6 female mice were initiated on experimental malnutrition diets (RBD or PD) or control diet (CD) immediately upon receipt from supplier. Challenge with 10<sup>6</sup> or 10<sup>7</sup> <i>Cryptosporidium parvum</i> oocysts, heat-inactivated <i>C</i>. <i>parvum</i> oocysts (Δ<i>C</i>. <i>parvum</i>) or PBS occurred via oral gavage 5 days after initiating diet. Serial weights were collected daily post-challenge, and fecal parasite shedding was determined by RT-PCR. On day 3–4 post-challenge, tissue parasite burden and mucosal injury was assessed by measuring ileum villus:crypt ratios and alterations in epithelial tight-junction proteins. (B) Impact of diet on growth (**<i>P</i><0.01, ***<i>P</i><0.001 <i>C</i>. <i>parvum</i><sup>PD</sup> vs. <i>C</i>. <i>parvum</i> or <i>C</i>. <i>parvum</i><sup>RBD</sup>), fecal parasite shedding (**<i>P</i><0.01 PD or RBD vs. CD day 2, *<i>P</i><0.05 PD vs. RBD or CD day 3), tissue parasite burden (*<i>P</i><0.05 PD vs CD or RBD ileum, *<i>P</i><0.05 PD vs CD colon), and ileum villus:crypt ratios (**<i>P</i><0.01 PBS<sup>CD</sup> vs PBS<sup>RBD</sup>, ***<i>P</i><0.001 PBS<sup>CD</sup> vs PBS<sup>PD</sup>, ****<i>P</i><0.0001 <i>C</i>. <i>parvum</i><sup>CD</sup> or <i>C</i>. <i>parvum</i><sup>RBD</sup> vs <i>C</i>. <i>parvum</i><sup>PD</sup>, ^<i>P</i><0.05 PBS<sup>CD</sup> vs. <i>C</i>. <i>parvum</i><sup>CD</sup>, <sup>##</sup><i>P</i><0.01 PBS<sup>PD</sup> vs. <i>C</i>. <i>parvum</i><sup>PD</sup> (n = 3-7/group). (C) Growth through three days post-challenge with either <i>C</i>. <i>parvum</i> or Δ<i>C</i>. <i>parvum</i>. (N = 5-10/group, **<i>P</i><0.01, ***<i>P</i><0.001 (<i>C</i>. <i>parvum</i> vs. either PBS or Δ<i>C</i>. <i>Parvum</i>) and fecal parasite shedding (n = 5-10/group, *<i>P</i><0.05, **<i>P</i><0.01). (D) Immunofluorescence staining of epithelial cell tight-junction proteins (ZO-1, occludin, claudin-2) in ileum of CD and PD-fed infected mice and uninfected controls (n = 4/group). (E) Dose dependent persistent growth faltering (n = 10/group, *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001, ****<i>P</i><0.0001 <i>C</i>. <i>parvum</i> 10<sup>7</sup> vs <i>C</i>. <i>parvum</i> 10<sup>6</sup>; and <sup>####</sup><i>P</i><0.0001 <i>C</i>. <i>parvum</i> vs PBS) and (F) fecal parasite shedding (**<i>P</i><0.01, ****<i>P</i><0.0001) through 21 days post-challenge. Data is representative of 2 replicate experiments.</p
