Or parasite loads in tissues. An analysis of variance (ANOVA) was conducted to account for the effects of relevant factors (inocula, day P.I.) and their interactions on daily oocyst excretion. Data analysis was performed with the statistical software Graphpad. Significance was defined as P,0.05.ResultsIn order to evaluate the infection susceptibility of mice Autophagy challenged with calibrated suspensions containing different intended doses, the amount of oocyst in feces was estimated periodically. All administered 1676428 doses containing different amounts of oocysts revealed to be infective for Dex-treated SCID mice but increasing the oocyst doses lead to an increase in the level of infectivity (P = 0.01). Two out of 7 mice (28.5 ) inoculated with an intended dose of 1 oocyst, 6/8 mice (75 ) receiving an intended dose of 10 oocysts, and all mice inoculated with intended doses of 102 and 105 viable oocysts developed chronic infection until euthanasia (45?00 days P.I.). None of the negative control mice discharged oocysts after oral inoculation with either PBS or 105 heat-inactivated oocysts (Table 1). The pattern of oocyst shedding of the different groups is shown in Fig. 1. At day 7 P.I. oocyst shedding was detected in mice from groups 2, 3 and 4 (challenged with 10, 100 and 105 oocysts) but not in animals from group 1 (challenged with one oocyst). Parasite shedding of animals from this latter group was detected for the first time after 15 days P.I. ANOVA analysis of the whole data set showed that the day P.I. and the inoculum size significantly influence the geometric means of oocyst sheding (P = 0.02 and 0.005, respectively): at 75 days P.I., mice inoculated with intended doses of 1, 10, 100 and 105 oocysts had a multiplication of 3.3, 3.25, 2.26 and 1.45 log respectively compared to initial inoculum. After 45 days post-infection, all groups of mice have a mean of oocyst shedding superior to 10,000 oocyst/g confirming the high proliferation rate of parasite growth at lower doses. After histological examination of tissues, gastrointestinal neoplastic lesions (Table 1) were observed in all Dex-treated SCID mice infected by C. parvum, whatever the inoculum (Figure 2). In the stomach, neoplastic lesions were Epigenetics localized in the antropyloric region and were detected as early as day 45 P.I. in all groups of infected mice. At day 45 P.I. these lesions were described as low grade intraepithelial neoplasia (LGIEN) or invasive adenocarcinoma for groups 1 and 2, and as high grade intraepithelial neoplasia (HGIEN) or invasive adenocarcinoma forgroups 3 and 4 (Table 1). At day 100 P.I., we observed the presence of adenocarcinoma of the submucosa invading the external muscularis layer in the antro-pylorique region of a mouse inoculated with a single oocyst (Figures 2A, 2B). In the ileo-caecal region, with intended doses of 1 and 10 oocysts a polypoid mucosa (adenoma) containing LGIEN and HGIEN lesions was observed respectively at 45 and 100 days P.I. (Figures 2C, 2D). With larger inoculum (100 oocysts) HGIEN was observed earlier (day 45 P.I.). Reticulin staining and cytokeratin immuno-labeling allowed the confirmation of the HGIEN: a fragmented basement membrane and neoplastic epithelial cells in the lamina propia were observed. These changes, which are typical of intramucosal adenocarcinoma, were observed in mice of group 3, after only 80 days P.I.. Neoplastic lesions seemed to be more severe in the stomach than in the ileo-caecal region. For quantitative analysis.Or parasite loads in tissues. An analysis of variance (ANOVA) was conducted to account for the effects of relevant factors (inocula, day P.I.) and their interactions on daily oocyst excretion. Data analysis was performed with the statistical software Graphpad. Significance was defined as P,0.05.ResultsIn order to evaluate the infection susceptibility of mice challenged with calibrated suspensions containing different intended doses, the amount of oocyst in feces was estimated periodically. All administered 1676428 doses containing different amounts of oocysts revealed to be infective for Dex-treated SCID mice but increasing the oocyst doses lead to an increase in the level of infectivity (P = 0.01). Two out of 7 mice (28.5 ) inoculated with an intended dose of 1 oocyst, 6/8 mice (75 ) receiving an intended dose of 10 oocysts, and all mice inoculated with intended doses of 102 and 105 viable oocysts developed chronic infection until euthanasia (45?00 days P.I.). None of the negative control mice discharged oocysts after oral inoculation with either PBS or 105 heat-inactivated oocysts (Table 1). The pattern of oocyst shedding of the different groups is shown in Fig. 1. At day 7 P.I. oocyst shedding was detected in mice from groups 2, 3 and 4 (challenged with 10, 100 and 105 oocysts) but not in animals from group 1 (challenged with one oocyst). Parasite shedding of animals from this latter group was detected for the first time after 15 days P.I. ANOVA analysis of the whole data set showed that the day P.I. and the inoculum size significantly influence the geometric means of oocyst sheding (P = 0.02 and 0.005, respectively): at 75 days P.I., mice inoculated with intended doses of 1, 10, 100 and 105 oocysts had a multiplication of 3.3, 3.25, 2.26 and 1.45 log respectively compared to initial inoculum. After 45 days post-infection, all groups of mice have a mean of oocyst shedding superior to 10,000 oocyst/g confirming the high proliferation rate of parasite growth at lower doses. After histological examination of tissues, gastrointestinal neoplastic lesions (Table 1) were observed in all Dex-treated SCID mice infected by C. parvum, whatever the inoculum (Figure 2). In the stomach, neoplastic lesions were localized in the antropyloric region and were detected as early as day 45 P.I. in all groups of infected mice. At day 45 P.I. these lesions were described as low grade intraepithelial neoplasia (LGIEN) or invasive adenocarcinoma for groups 1 and 2, and as high grade intraepithelial neoplasia (HGIEN) or invasive adenocarcinoma forgroups 3 and 4 (Table 1). At day 100 P.I., we observed the presence of adenocarcinoma of the submucosa invading the external muscularis layer in the antro-pylorique region of a mouse inoculated with a single oocyst (Figures 2A, 2B). In the ileo-caecal region, with intended doses of 1 and 10 oocysts a polypoid mucosa (adenoma) containing LGIEN and HGIEN lesions was observed respectively at 45 and 100 days P.I. (Figures 2C, 2D). With larger inoculum (100 oocysts) HGIEN was observed earlier (day 45 P.I.). Reticulin staining and cytokeratin immuno-labeling allowed the confirmation of the HGIEN: a fragmented basement membrane and neoplastic epithelial cells in the lamina propia were observed. These changes, which are typical of intramucosal adenocarcinoma, were observed in mice of group 3, after only 80 days P.I.. Neoplastic lesions seemed to be more severe in the stomach than in the ileo-caecal region. For quantitative analysis.