Colin F. Mills

Biochemical and pathological changes in tissues of Friesian cattle during the experimental induction of copper deficiency.

1. Copper deficiency was induced in five Friesian cattle offered a semi-synthetic diet containing less than 1 mgCu/kg. Changes in blood and liver Cu contents and in the Cu-containing enzymes, ferroxidase I (caeruloplasmin; EC 1.16.3.1) and monoamine oxidase (EC 1.4.3.4) of plasma and cytochrome oxidase (EC 1.9.3.1) of liver and skeletal muscle were monitored during Cu depletion. 2. Rapid decreases in blood and liver Cu and plasma ferroxidase I activity were found at least 80 d before the first appearance of overt clinical signs of deficiency. Plasma monoamine oxidase and liver cytochrome oxidase activities decreased less rapidly and thus may provide useful indices of chronic Cu depletion.

Protective effect of zinc supplementation against copper toxicosis in sheep

1. A study has been made of the effects of dietary zinc supplementation on the development of copper toxicosis in three groups each of eight 12-week-old lambs. 2. None of the lambs receiving 420 mg Zn/kg diet developed Cu toxicosis in the 24-week experimental period, compared with three in the control group receiving 43 mg Zn/kg and possibly one in the group receiving 220 mg Zn/kg. 3. Liver Cu concentrations were reduced by up to 40% in the Zn-supplemented animals, with concomitant reductions, especially in the early stages of the experiment, in the extent of liver damage, as assessed by measurement of plasma aspartate aminotransferase (EC 2.6.1.1) and arginase (EC 3.5.3.1) activities. 4. Plasma and liver Zn concentrations were increased only slightly in the lambs receiving the Zn-supplemented diets, and the only indication of possible toxic effects of the Zn supplements was the development of a slight anaemia in those animals receiving 420 mg Zn/kg diet. 5. The results suggest that the incidence of Cu toxicosis in sheep may be controlled by increasing their dietary Zn intake.

Copper and molybdenum absorption by rats given ammonium tetrathiomolybdate

Previous studies have shown that the tetrathiomolybdate ion [MoS4(2-)] is a potent antagonist of Cu metabolism. Effects of orally administered MoS4(2-) on the absorption and tissue distribution of 64Cu in rats have now been investigated. Four or 12 mg Mo/kg diet, when given as MoS4(2-), strongly inhibited 64Cu absorption and modified the fate of absorbed Cu, decreasing hepatic and renal uptake but increasing plasma retention of 64Cu. These effects were not induced by equivalent dietary concentrations of Mo as MoO4(2-) or when S2- was given as CaS. Clinical and biochemical effects induced by orally administered MoS4(2-) were abolished by increasing dietary concentrations of Cu. Such treatment also inhibited the absorption and tissue retention of 99Mo derived from 99MoS4(2-). Intraperitoneal administration of Cu ameliorated clinical effects attributable to MoS4(2-) but neither inhibited 99Mo absorption nor the appearance of systemic defects in Cu metabolism. Since the absorption of MoS4(2-) (or its derivatives) from the gastrointestinal tract is inhibited by Cu, it is evident that the site of its action as an antagonist influencing either the absorption or the subsequent metabolic fate of Cu depends upon the ratio Cu/MoS4(2-) in the diet.

Effects of molybdate, sulfide, and tetrathiomolybdate on copper metabolism in rats

Species differences in the response to dietary MoO4(2)- as a metabolic antagonist of Cu are considered briefly. Suggestions that (i) the potency of MoO4(2)- as a Cu antagonist is enhanced by normally innocuous dietary concentrations of S20 and (ii) that MoS4(2)- may be a more effective antagonist than either MoO4(2)- or S2- were investigated in a series of studies with rats. Diets including MoS4(2)- but not of MoO4(2)- or S2- alone promoted a decline in hepatic Cu and ceruloplasmin activity and induced clinical signs of Cu deficiency. Evidence of concurrent anomalies in the partition of Cu between tissues and in the distribution of Cu between proteins of plasma and kidney cytosol suggested that such effects were partly attributable to the development of systemic defects in Cu metabolism. The relationship of such findings to the suggested involvement of MoS4(2)- or its derivatives in the etiology of Mo-induced Cu deficiency in ruminant animals is considered.

Copper metabolism in rats given di- or trithiomolybdates

As one of a series of studies on the influence of S substitution of MoO4(2-) on the action of Mo as an antagonist of Cu, the effects of MoO2S2(2-) and MoOS3(2-) on the metabolism of Cu by rats have been investigated. Administration of either oxythiomolybdate increased both plasma Cu concentration and the retention of Cu by the albumin fraction of plasma. In contrast to the effects of MoS4(2-), described previously, neither oxythiomolybdate inhibited 64Cu absorption from the digestive tract or induced biochemical or clinical signs of Cu deficiency. Evidence is discussed suggesting that the potency of the thio- and oxythiomolybdates as inhibitors of Cu utilization is directly proportional to the degree of thio substitution within the series series MoO4- chi S chi 2-.

Copper deficiency and tissue glutathione concentration in the rat

Abstract Copper deficiency in rats increased renal vein and arterial (heart) plasma GSH concentration by approximately 50%. There was no change in plasma GSSG concentration. Renal vein plasma GSSG/GSH ratio was decreased in copper deficiency, which is consistent with previous reports showing a copper-dependant thiol oxidase activity in the renal basement membrane. No change occurred in arterial plasma GSSG/GSH ratio. Hepatic GSH concentrations were also elevated by 50% in copper deficiency, GSSG concentrations were unaffected, but GSSG/GSH ratio was depressed. Renal and cardiac tissue GSH and GSSG were unaffected by copper deficiency. The decreased SOD activity and GSH-Px activity observed in copper deficiency may contribute to increased hepatic and plasma GSH concentrations.

Glutathione peroxidase activity and erythrocyte stability in calves differing in selenium and vitamin E status

1. Glutatione peroxidase activity (EC 1.11.1.9) and erythrocyte stability were measured in Friesian bull calves which were given for 36 weeks semi-purified diets either adequate or low in selenium or vitamin E or both. 2. Dietary Se or vitamin E content had no effect on growth rate and haematlogical values. None of the calves exhibited clinical deficiency symptoms and serum aspartate amino transferase (EC 2.6.1.1) and creatine phosphokinase (EC 2.7.3.2) activities remained normal. Heart and skeletal muscles of all calves appeared macroscopically and microscopically normal ato autopsy. 3. Glutathione peroxidase activity in plasma, blood and other tissues, except the testis, was significantly lower in calves receiving low dietary Se but was independent of dietary vitamin E content. 4. Plasma vitamin E levels decreased rapidly and to very low levels in calves given low vitamin E diets irrespective of the Se content of the diet. 5. A low dietary vitamin E intake increased the susceptibility of erythrocytes to auto- and peroxidative haemolysis whereas a low Se intake in the presence of adequate vitamin E did not. However, erythrocytes from calves receiving low Se and low vitamin E were more susceptible to peroxidative haemolysis than erythrocytes from calves receiving low vitamin E and adequate Se. The effect of dietary vitamin E content on osmotic haemolysis induced by hypotonic saline was variable. 6. The results suggest that measurement of blood glutathione peroxidase activity and the susceptibility of erythrocytes to auto- or peroxidative haemolysis could be used for the differential diagnosis of subclinical Se and vitamin E deficiency in ruminants.

Nutritional Aspects of Molybdenum in Animals

Publisher Summary This chapter reviews the nutritional aspects of molybdenum in animals. The evidence of functional roles for molybdenum in xanthine oxidase, aldehyde oxidase, and sulphite oxidase suggests that Mo is an essential trace nutrient for animals. Nevertheless, there are few instances in which a dietary deficiency of this element has been directly implicated as a cause of poor growth and fewer still in which it has been recognized to be the cause of metabolic defects provoking the appearance of characteristic pathological lesions. The assumption that Mo deficiency was unlikely to be of great economic significance in the nutrition of farm animals or to influence human health has resulted in much less effort being devoted to the study of the metabolic and pathological consequences of Mo deficiency in animals than in plants or bacteria. Because of its significance in the nutrition of ruminant animals, much greater emphasis has been placed upon the investigation of the importance of Mo as an antagonist of copper and, thus, its role in the etiology of Cu deficiency. In the instance of Mo deficiency, the onus is upon the nutritionist to utilize newly developed techniques, first, to reinvestigate quantitative requirements for Mo and, second, to produce appropriate material for biochemical and pathological studies.

Stimulation of peroxidation in rat liver microsomes by (copper, zinc)-metallothioneins.

The abilities of pig liver (copper, zinc) metallothionein I and rat liver zinc metallothionein II to modify lipid peroxidation in incubations of liver microsomes have been compared with the activities of reduced glutathione, mannitol, quinacrine, EDTA, dimethyl-pyrroline-N-oxide and phenyl-butyl-nitrone. Lipid peroxidation was determined by assay of thiobarbituric acid reactive substance formation in incubations of microsomes with iron/ADP or a mixture of xanthine and xanthine oxidase. Zinc metallothionein II had no effect on the extent of peroxidation in either system but (copper, zinc) metallothionein I caused a stimulation of peroxidation initiated by xanthine and xanthine oxidase, all other compounds tested were inhibitory. Gel exclusion chromatography of incubations of (copper, zinc) metallothionein I with xanthine and xanthine oxidase revealed aggregation of the metalloprotein. This may have exposed copper in a form capable of initiating peroxidation.

Effects of tetrathiotungstate and dithiotungstate on copper metabolism in rats

The effects of dietary supplementation with ammonium tetrathiotungstate and dithiotungstate on copper metabolism in young rats have been investigated. The addition of WS4(2-) (4-32 mg W/kg diet) decreased growth rates and induced clinical and biochemical signs of Cu deficiency. These were overcome by increasing the dietary content of Cu. The intestinal absorption of 64Cu was impaired and the tissue distribution of absorbed Cu modified by the administration of WS4(2-). No clinical or biochemical defects indicative of Cu deficiency developed in rats receiving WO2S2(2-) in their diet at concentrations up to 32 mg W/kg. In contrast, this oxythioanion enhanced the content of Cu in plasma liver and kidney, especially when dietary Cu was increased. Most of the additional Cu retained by plasma and kidney was associated with albumin and metallothionein, respectively. The effects of these W sources are compared with those caused by their thio- and oxythiomolybdate analogs and are discussed in relation to the mechanisms whereby Mo induces Cy deficiency in ruminants.