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No deaths occurred when intoxicated mice were treated with 80 µg of polyclonal goat anti-ricin antibodies by 6 h post-intoxication (Fig. 4B) but about 30% lethality were observed when antibodies were applied at 7 h post-intoxication.
Some lethality was observed when antibodies were added at 7 h post-intoxication, indicating that for some mice, enough ricin reached the bloodstream and was absorbed into tissues between 6 and 7 h post intoxication, before antibody administration.
Decreased protection was observed when antibodies were applied post-ricin, even as early as 2 min post-intoxication.
Our detailed measurement of ricin content in sera at early time points post-intoxication revealed a more complete picture of toxin toxicokinetics.
The samples of cardiac blood, urine, gastric and duodenal wall, liver, lung, kidney, heart and diaphragm, showed quantifiable levels of PQ even at 6 days post-intoxication.
100 µg of polyclonal goat anti-ricin antibodies diluted in PBS buffer were introduced iv at 30 min prior to toxin or at indicated time intervals post-intoxication.
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Intoxicated mice were protected from death when antibodies were introduced up to 6 h post- intoxication (Fig. 4B), suggesting that it took at least 6 h to traffic significant amount of ricin from the mouse stomach to the bloodstream.
At higher doses (up to1 mg/mouse), the presence of ricin was clearly found in feces deposited within the first 2 hours after intoxication, while at lower doses (400 µg/mouse or less), ricin was detected frequently in feces deposited after 2 hours post intoxication.
Botulinum neurotoxin serotype A and NAPs in the toxin complex were detectable inside intestinal cells beginning at 2 h post intoxication.
At 2, 3, 4, 6, 7, 8 and 24 h post intoxication, three mice per time point were euthanized and perfusion-fixed with normal saline followed by 4% PFA.
When mice were treated with BoNT/A complex, there was binding of BoNT/A to the outer layers of the intestinal epithelia as early as 2 h and a gradual increase in BoNT/A staining in the lamina propria and intestinal crypts over time, peaking at 6 h post intoxication.
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