Absorption, tissue distribution, and elimination of residues after 2,4,6-trinitro[14C]toluene administration to sheep

The compound 2,4,6-trinitrotoluene (TNT) is a persistent contaminant of some industrial and military sites. Biological bioremediation techniques typically rely on the immobilization of TNT reduction products rather than on TNT mineralization. We hypothesized that sheep ruminal microbes would be suitable for TNT destruction after phytoremediation of TNT-contaminated soils by cool-season grasses. Therefore we investigated the fate of [14C]TNT in ruminating sheep to determine the utility of ruminant animals as a portion of the bioremediation process. Three wether sheep were dosed with 35.5 mg each of dietary unlabeled TNT for 21 consecutive days. On day 22 sheep (41.9 +/- 3.0 kg) were orally dosed with 35.5 mg of [14C]TNT (129 microCi; 99.1% radiochemical purity). Blood, urine, and feces were collected at regular intervals for 72 h. At slaughter, tissues were quantitatively collected. Tissues and blood were analyzed for total radioactive residues (TRR); excreta were analyzed for TRR, bound residues, and TNT metabolites. Plasma radioactivity peaked within 1 h of dosing and was essentially depleted within 18 h. Approximately 76% of the radiocarbon was excreted in feces, 17% in urine, with 5% being retained in the gastrointestinal tract and 1% retained in tissues. Parent TNT, dinitroamino metabolites, and diaminonitro metabolites were not detected in excreta. Ruminal and fecal radioactivity was essentially nonextractable using ethyl acetate, acetone, and methanol; covalent binding of fecal radioactive residues was evenly distributed among extractable organic molecules (i.e., soluble organic matter, soluble carbohydrate, protein, lipid, and nucleic acid fractions) and undigested fibers (cellulose, hemicellulose, and lignin). This study demonstrated that TNT reduction within the ruminant gastrointestinal tract leads to substantial immobilization of residues to organic matter, a fate similar to TNT in other strongly reducing environments.