5 cm are then formed and injected with 400 u03bcl of 50 mm tris buffer ph 7 4 or with tris buffer containing c difficile culture filtrate 20 ug of protein the inhibitory effect of secretory iga is assessed by the addition of secretory iga 200 ug to the toxins prior to injection into the ileal lumen the abdominal incision is closed and anesthesia maintained with sodium pentobarbital the animals are sacrificed after 4 hours and the ileal loops immediately harvested loop weight to length ratio is determined as a measure of enterotoxin effect mannitol excretion indicating intestinal permeability is measured by counting radioactivity in the luminal fluid ileal tissue samples are also fixed in formalin paraffin embedded and sections stained with hematoxylin and eosin the histologic severity of enteritis is graded taking into account the following features i neutrophil margination and tissue infiltration ii hemorrhagic congestion and edema of the mucosa iii epithelial cell damage a score of 0 to 3 denotes increasingly severe pathological changes treatment of a person ill with c difficile associated disease with secretory iga an adult individual ill with c difficile associated disease is treated with secretory iga containing antibody activity against c difficile toxin treatment is with 1 gram orally three times daily together with vancomycin in appropriate dosage treatment is continued until symptoms resolve and the stool becomes negative for c difficile toxin bacon a e 3rd fekety r lrnrunoglobulin g directed against toxins a and b of clostridium difficile in file general population and patients with antibiotic.