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1. The ability of organic and inorganic sulphur to influence the copper and molybdenum metabolism of sheep was compared in a series of three 2 X 2 factorial design experiments. In each experiment, four groups of five to seven hypocupraemic ewes were repleted with a basal diet supplemented with 6 mg Cu/kg and containing S and Mo at one of two concentrations, 1 or 4 g S and 0-5 or 4-5 mg Mo/kg respectively. Sodium sulphate (Expt I), methionine (Expt 2) or cysteine (Expt 3) were used as the S sources. Cu and Mo concentrations in plasma were estimated in each experiment and in Expt 3 the concentrations of Cu in liver and Mo in urine were also estimated. 2. The effects of the three S sources on Cu and Mo metabolism were similar. Repletion of the plasma Cu pool was unaffected by Mo alone, reduced by S alone and totally inhibited by Mo+S. Plasma Mo was greatly increased by Mo supplements, slightly decreased by S supplements and unaffected by Mo and S supplements given together. 3. In Expt 3 the treatments were found to affect urinary Mo and plasma Mo in a similar manner; S prevented dietary Mo from increasing Mo excretion. The only group to show a significant repletion of the liver Cu pool was that given Mo alone. 4. Supplementation of the diet with organic S significantly reduced the within-treatment variation in plasma Cu and Mo. liver Cu and urinary Mo. 5. It is suggested that variations in dietary S and Mo within the normal range for herbage affect the Cu and Mo metabolism of the grazing animal, and that total S rather than inorganic S is the more useful measurement in the context of the Cu-Mo-S interrelationship.

1. Protein-bound, trichloracetic acid- (TCA) insoluble 99Mo appeared in plasma a few hours after the introduction of 99Mo-labelled molybdate (30 mg Mo) into the rumen of sheep maintained on a basic diet supplemented with elemental sulphur (3 g S/d). 2. Most of the 99Mo could be displaced from its protein carrier in vitro and the labelled compounds displaced were identified by sephadex G-25 chromatography as di- and trithiomolybdate. Tetrathiomolybdate was not detected. 3. In control experiments protein-bound, TCA-insoluble 99Mo predominated in plasma after the direct administration of [99Mo]tetrathiomolybdate, either into the rumen or intravenously. The 99Mo could be displaced in vitro and [99Mo]tetrathiomolybdate identified, although [99Mo]trithiomolybdate was also present. The study provides direct evidence of thiomolybdate synthesis and absorption in ruminants in vivo.

1. At 16 h after the rapid injection of 99Mo-labelled compounds into the rumen of sheep maintained on dried grass (6.2 mg molybdenum/kg dry matter (DM), 4.3 g sulphur/kg DM), labelled thiomolybdates associated with the digesta solids or bound to plasma macromolecular species were displaced from their carriers in vitro and identified by Sephadex G25 chromatography. 2. After molybdate injection, the thiomolybdates displaced from rumen, duodenal and ileal solids were predominantly the trithio- and tetrathio- species. Dithiomolybdate was present to a relatively minor extent. Trace amounts of di- and trithiomolybdates were detected in the liquid phase of digesta from the duodenum. 3. Whether injected into the rumen as molybdate or tetrathiomolybdate, bound 99Mo appearing in plasma was present mainly as di- and trithio- species. The tetrathio- species appeared in trace amounts in plasma only after tetrathiomolybdate injection, despite its existence almost exclusively in this form in rumen digesta. 4. The present study provides direct evidence for thiomolybdate synthesis within the rumen and indicates that while the effects of thiomolybdates in inhibiting copper absorption are likely to be due to tri- and tetrathiomolybdates, post-absorptive effects on Cu metabolism are probably due to di- or trithiomolybdate.

Sheep were infused intravenously with 99Mo- and 35S-labelled tri- and tetrathiomolybdates (1-2 mg Mo). Most of the plasma radioactivity was trichloroacetic acid (TCA)-insoluble after infusion, but the stability of this fraction was reduced by pre-infusion or subsequent infusion with unlabelled thiomolybdates. Most of the 99Mo and 35S was shown to be associated with albumin. It was concluded that compounds bound to albumin were relatively stable, but displaced or unbound thiomolybdates were rapidly hydrolysed to molybdate and sulphate. There was no evidence of an irreversible interaction of either 35S or 99Mo with copper in plasma, despite the appearance of a TCA-insoluble Cu fraction. Increased dietary Cu did not increase the retention of 35S in plasma or affect the exchangeability of 36S-labelled thiomolybdates bound to albumin.

1. A 30-week study has been made in growing ewe lambs of the effects of dietary supplementation with molybdenum, or Mo plus inorganic sulphate on the distribution of copper in their blood plasma and kidneys. 2. The addition of 25 mg Mo and 5 g SO42-/kg diet increased Cu concentrations in plasma and kidney and decreased those in liver. Plasma caeruloplasmin activities (ferroxidase; EC 1.16.3.1) were unaffected. 3. Subcellular fractionation of the kidney cortex and gel filtration of the plasma and kidney cortex cytosol showed that the increased Cu content of these tissues was associated with abnormalities in the distribution of Cu. It appeared that both Cu and Mo were associated with the same proteins. 4. Dietary supplementation with Mo alone (25 mg/kg) had no effect on plasma or kidney Cu concentrations, suggesting that S metabolism is involved in the formation of the abnormal Cu-binding proteins in plasma and kidney. 5. The results are discussed in relation to the induction of Mo-induced Cu-deficiency states and the possible involvement of thiomolybdates.