The role of metallothionein and zinc in hepatic copper accumulation in cattle

The role of metallothionein (MT) and zinc (Zn) in hepatic copper (Cu) accumulation in calves from a region in North-West Spain has been investigated. In this region there is intensive pig farming, and animals with liver Cu concentrations above normal are not uncommon. Concentrations of hepatic MT were not related to Cu accumulation but were strongly dependent on the Zn status of the animal. When analysing the metal content bound to MT it was observed that Cu-MT values, in the same way as Zn-MT, were directly correlated with MT concentrations in the liver, indicating that although Cu is a poor inducer of MT synthesis, it can compete with Zn for MT binding sites. The ability of Cu to displace Zn from MT is highly dependent on the Cu:Zn ratio in the cell, Cu being the main metal in MT at the higher levels of Cu exposure in Galician cattle. In spite of this, the percentage of the total hepatic Cu bound to MT is very low in these animals, indicating that cattle have a very limited capacity to accumulate Cu-MT in the liver, and may therefore have a greater susceptibility to Cu toxicosis.     

Clinical observations of cattle and buffalos with experimentally induced chronic copper poisoning

The susceptibility of cattle and buffalos to chronic copper poisoning (CCP) was compared by using cattle (n = 10) and buffalo (n = 10) steers distributed into two copper supplemented (n = 6) and two control (n = 4) groups. Supplemented animals received 2 mg copper (Cu)/kg body weight daily for one week, with an additional 2 mg weekly until the end of the experiment (day 105). Three liver biopsies (day 0, 45, and 105) were obtained for mineral analyses; clinical examinations and blood samples were obtained every 15 days. Three supplemented cattle and two buffalos with typical manifestations of CCP died. There were no differences in the frequency of mortality between cattle and buffalos; hepatic copper concentration was higher in cattle than buffalos. These findings suggest that buffalos and cattle might be equally susceptible to CCP. However, buffalos accumulate less liver copper than cattle and have a lower threshold of hepatic Cu accumulation, which leads to clinical manifestation of CCP.     

Influence of copper supplementation on blood and liver copper in cattle fed tall fescue or quackgrass

Copper concentrations were measured in naturally Cu-deficient and Cu-supplemented yearling Hereford heifers fed tall fescue (Festuca arundinacea Schreb.) or quackgrass (Agropyron repens [L] Beauv.) forage. The data were used to establish a relationship between dietary Cu supplementation and accumulation of liver Cu, and to test the influence of diet and Cu supplementation on blood plasma Cu and ceruloplasmin oxidase activity. Dietary Cu supplementation was positively correlated with accumulation of liver Cu; the rate of increase in liver Cu levels diminished at high supplementation rates (liver Cu = 410.43 - 353.47 x e-.00295 x mg Cu supplement; R2 = .740), suggesting an efficient mechanism preventing fast, toxic Cu-overloading in cattle. The linear relationship (R2 = .7878) between blood plasma Cu and ceruloplasmin oxidase activity indicated that similar percentages of blood plasma Cu were present as ceruloplasmin in deficient, normal and Cu-supplemented cattle. However, the portion of plasma Cu occurring as ceruloplasmin changes significantly with type of feed. When compared with quackgrass, tall fescue not only produced rapid Cu depletion in unsupplemented cattle, but affected intermediary Cy metabolism and activity of Cu-enzyme ceruloplasmin differently at all levels of experimental Cu supplementation.     

Intracellular distribution of copper and zinc in the liver of copper-exposed cattle from northwest Spain

The intracellular distribution of copper (Cu) and zinc (Zn) in the livers of normal and moderately Cu-exposed cattle was investigated with the aim of improving understanding of the pathophysiology of Cu accumulation in cattle. At total liver Cu concentrations within the generally accepted normal range (25-100 mg/kg fresh weight) the large-granule fraction was the main subcellular compartment for Cu accumulation, followed by the cytosol and the nucleus, whereas in the microsomal fraction Cu accumulation was very low. With increasing Cu exposure, the capacity of the large-granule fraction to accumulate Cu decreased, proceeding towards a plateau (estimated at about 80 mg/kg at a projected total liver Cu concentrations of 450 mg/kg), accompanied by progressively greater Cu accumulation in the nucleus and cytosol. Total liver Cu concentration had little influence on subcellular Zn distribution, with hepatic Cu concentrations being only moderately correlated with Zn concentration in the large-granule fraction. There was a strong association between the proportion of total Cu bound to metallothionein (MT) and the proportion of total Zn in the large-granule fraction, suggesting that Zn displaced from MT is taken up by the lysosomes for excretion. This pattern of Cu accumulation, as in sheep, may be due to the limited capacity for metallothionein binding of Cu and excretion in bile.     

Effect of calcium supplements to a maize-soya diet on the bioavailability of minerals and trace elements and the accumulation of heavy metals in growing rats

Thirty-five (five groups with seven animals each) male albino rats (initial average weight = 44 g) were fed phytate-rich diets (analysed phytic acid concentration = 6.9 g/kg) based on maize and soy bean meal (5 g Ca, 3 g P, 1.2 g Mg, 23 mg Zn, 10 mg Pb, 5 mg Cd/kg diet). Experimental diets were supplemented with 0, 2, 4, 6 and 8 g calcium from CaCO3 per kg. The supplementation of increasing amounts of calcium resulted in a dose-dependent decrease in the apparent absorption of phosphorus. Furthermore, apparent zinc absorption and femur zinc concentration were moderately decreased due to the calcium supplementation. Kidney Cd concentration was significantly lower in rats that were fed the high calcium diets in comparison with the control animals. Femur lead concentration and hepatic delta-aminolevulinic acid dehydratase, which are known to be sensitive parameters of lead accumulation remained unchanged by the different dietary treatments. Magnesium absorption as well as liver and plasma zinc concentration and activity of plasma alkaline phosphatase were also unaffected. Although calcium supplementation may lead to a decrease in the accumulation of certain heavy metals such as cadmium, the carry-over of lead was not affected under the given experimental conditions. Furthermore, calcium-phytate-zinc interactions may adversely affect zinc bioavailability in growing rats.     

Evaluation of a commercially available molybdate formulation and zinc oxide boluses in preventing hepatic copper accumulation and thus enzootic icterus in sheep

The efficacy of a molybdate formulation and a zinc oxide bolus as prophylactic agents for enzootic icterus was evaluated in sheep. Before copper loading, liver biopsies were performed on 12 male, 6-month-old, Mutton Merino sheep to determine hepatic copper (Cu) and zinc (Zn) concentrations. The animals were restrictively randomised according to liver copper concentrations to 3 treatment groups (n = 4) to achieve similar mean liver copper concentrations per group. All sheep received 4 ml/kg of a 0.5 % aqueous solution of CuSO4 5H2O intraruminally 7 days per week for 10 weeks. On Day 0 the sheep in the Mo-group were injected subcutaneously with 42 mg molybdenum (Mo) contained in a commercial molybdate formulation. The animals in the Zn-group each received a zinc oxide bolus, containing 43 g zinc oxide, via a rumen cannula. Treatment was repeated on Day 42. Four animals served as untreated controls. Urinary copper excretion, plasma copper concentration, haematocrit and glutamate dehydrogenase (GLDH) activity were determined throughout the trial. The animals were sacrificed after 10 weeks and liver samples were submitted for histopathological examination. Liver and kidney copper and zinc concentrations were determined. Neither the molybdate treatment nor the zinc oxide boluses prevented hepatic copper accumulation. The urinary copper excretion, plasma copper concentration, haematocrit and GLDH activity were not significantly different (P > 0.05) from the controls.     

Breed differences in sheep with respect to the accumulation of copper in the liver

Male lambs of the pure breeds Texel, Flemish milk sheep, and Finnish landrace, and of the crossbrech Texel x Friesian milk sheep, Texel x Finnish landrace, Suffolk x Friesian milk sheep, and Finnish landrace x He de France were fed hay and concentrates, the latter differing only in the content of copper. At high (± 35 ppm) and at moderately low (± 10 ppm) copper levels in the concentrate, breed differences in the accumulation of copper in the liver were studied‐Hepatic Cu‐levels increased significantly in the case of the high Cu‐fed animals, but great differences in the metabolism of excessive amounts of dietary Cu exist between the particular breeds and crossbreds. The Texel breed was most susceptible to an excess of dietary Cu and the Finnish landrace was the least susceptible breed, while the Flemish milk sheep and the crossbreds were intermediate. In blood serum, the enzyme activities of sorbitol dehvdrogenase, lactate dehydroge‐nase, γ‐glutamyl transferase, and glutamic oxaloacetic transaminase were increased to a greater or less extent in the high Cu‐fed animals, but the correlation with hepatic Cu‐content was poor. For the most susceptible breeds, even a level of 10 ppm Cu in a ration with nearly equal amounts of hay and concentrate is insufficiently low for the prevention of Cu‐accumulation in the liver.     

The assessment of blood copper status in cattle: a comparison of measurements of caeruloplasmin and elemental copper in serum and plasma

AIM: To evaluate the effect of test, either copper (Cu) concentration or caeruloplasmin (CP) activity, and sample type, either serum or plasma, on the diagnosis of blood Cu status in cattle. METHODS: Paired serum and heparinised plasma samples taken from 125 cattle in 13 herds were tested for Cu concentration and CP activity. The individual results for serum Cu concentration and serum and plasma CP activities were compared with the plasma Cu concentration results, as were their diagnostic values as determined by reference ranges, i.e. 'marginal', 'adequate', 'excess'. RESULTS: The overall mean serum Cu concentration was 2.92 micromol/L lower than the mean plasma Cu concentration; however, there was significant variability between individual samples, and the 95% limits of agreement ranged from 0.44 micromol/L more to 6.28 micromol/L less. The relationship between CP activity and plasma Cu concentration was less variable; the 95% prediction interval for plasma Cu concentration from CP activity was +/- 2.8 micromol/L, and was unaffected by whether CP activity was measured in plasma or serum. Using the threshold currently recommended for 'marginal' status of <8.0 micromol/L for serum Cu concentration identified a significantly different population of cattle than a threshold of <9.0 micromol/L for plasma samples. Altering the threshold to <7.0 micromol/L for serum Cu concentration produced better agreement. For CP activity, a threshold of 15 IU/L for both serum and plasma identified the same population as a threshold of <9 micromol/L for plasma Cu concentration. CONCLUSIONS: Serum Cu concentration is not a suitable substitute for plasma Cu concentration for the detection of 'marginal' blood Cu status in cattle as the individual variability in the apparent loss of Cu during clotting is too great. In this study, CP activity, in both serum and plasma, was found to be a suitable substitute for the detection of 'marginal' blood Cu status. CLINICAL RELEVANCE: The use of serum Cu concentration rather than plasma Cu concentration in the diagnosis of Cu responsive disease in cattle needs to be re-evaluated as does the way in which individual sample results are used in such tests.     

Copper,Zinc and Molybdenum in Livers of Norwegian Cattle At Slaughter

The concentrations of copper, zinc and molybdenum were measured in samples of cattle liver from 10 slaughter-houses in Norway. A total of 335 samples were analysed. A clear accumulation of copper with age was found, the average copper level in the younger animals (≦ 3 years, n = 194) being 30 µg Cu/g liver wet weight, and in the older ones (> 3 years, n = 141) 59 µg Gu/g. The range in the copper values found was considerable, though significant differences between some of the districts were recorded. Copper concentrations were classified as low (≦10 µg Gu/g) in 9.6 % of the samples. Zinc showed no accumulation with age, nor were there any differences in zinc levels found in animals from different districts, the average level being 32 µg Zn/g liver wet weight. The picture was the same for molybdenum, no differences between age groups or districts being found. The average level was 1.0 µg Mo/g liver. There was no significant correlation between levels of copper, zinc or molybdenum.The supply of copper and zinc to cattle in Norway seems close to sufficient, but copper- and zinc-fortified mineral supplementation of cattle feed is still to be recommended. There seems to be no need for molybdenum supplementation in cattle.     

Iron storage disease in captive Egyptian fruit bats (Rousettus aegyptiacus): relationship of blood iron parameters to hepatic iron concentrations and hepatic histopathology.

This study evaluated the relationship between blood iron parameters and hepatic iron concentrations, and correlation of histologic findings with hepatic iron concentrations in a captive population of Egyptian fruit bats (Rousettus aegyptiacus) and island flying foxes (Pteropus hypomelanus). Blood samples were collected for complete blood counts, plasma biochemical profiles, serum iron concentrations, total iron-binding capacity, whole-blood lead concentrations, and plasma ferritin assays. Liver samples obtained by laparotomy were divided, with one half processed for histologic examination and the other half frozen and submitted for tissue mineral analysis. The histologic sections were scored by two blinded observers for iron deposition, necrosis, and fibrosis. The Egyptian fruit bats had significantly higher liver iron (mean = 3,669 +/- 1,823 ppm) and lead (mean = 8.9 +/- 5.8 ppm) concentrations than the island flying foxes (mean [Fe] = 174 +/- 173 ppm, mean [Pb] = 1.9 +/- 0.5 ppm). Hepatic iron concentrations significantly correlated with tissue lead concentrations, histologic grading for iron and necrosis, serum iron, transferrin saturation, and plasma ferritin (P < 0.001). Blood lead concentrations negatively correlated with tissue lead concentrations (P < 0.001). When the product of transferrin saturation and serum iron was greater than 51, an individual animal had a high probability of having iron overload. When the product of these two variables was greater than 90, there was a high probability that the animal had hemochromatosis. On the basis of this study, it appears that evaluation of serum iron, transferrin saturation, and plasma ferritin are useful and noninvasive methods for diagnosis of hemochromatosis in Egyptian fruit bats.     

Growth response, blood characteristics and copper accumulation in organs of broilers fed on diets supplemented with organic and inorganic dietary copper sources

1. A 56-d experiment was conducted to study the comparative influence of organic and inorganic dietary copper (Cu) sources on growth, blood characteristics and copper accumulation in organs of broilers. 2. A total of 480 Arbor-Acre unsexed broilers were fed on diets containing copper sulphate (CuSO(4)) or copper proteinate (Cu Pro) at concentrations of 50, 100 or 150 mg/kg of Cu supplementation. The birds were given a broiler starter diet from 1-28 d and a broiler finisher diet from 29-56 d which contained 30·8 mg/kg and 41·1 mg/kg basal copper concentration respectively. Growth performance, blood characteristics and Cu accumulation in organs of the broilers were measured. 3. At 28 d, Cu Pro-fed birds had improved feed conversion ratio compared with CuSO(4). At 56 d, birds fed on Cu Pro diets had significantly greater body weight than CuSO(4)-fed birds. Birds fed on CuSO(4) supplemented diets had significantly better feed conversion efficiency. Feed consumptions for the two Cu sources were not significantly different. At no stage did the concentration of added Cu affect the productive traits measured. 4. Cu Pro supplementation increased haemoglobin concentration but reduced plasma triglyceride and plasma cholesterol. Plasma cholesterol decreased as Cu concentration increased. 5. There was a greater accumulation of Cu in the blood, heart, lung, liver and bone of broilers fed on Cu Pro than in those receiving CuSO(4). The liver Cu concentration increased as dietary Cu concentration increased. 6. Cu Pro was more effective in promoting growth and reducing blood cholesterol, and was more bio-available in the organs of broilers.     

The effect of copper-amended fertiliser and copper oxide wire particles on the copper status of farmed red deer (Cervus elaphus) and their progeny

AIM: To determine changes in serum and liver copper concentrations in postnatal, weaner, yearling, and mature deer after grazing pasture topdressed with copper (Cu) at two rates of application of copper sulphate (CuSO4(.)5H2O), and following oral administration of copper oxide (CuO) wire particles to some of the deer. METHODS: In mid-March 2000 (Year 1), 1.1-ha paddocks (two/treatment) of ryegrass/white clover pasture received either 0 (Control), 6 (Low) or 12 (High) kg CuSO4(.)5H2O /ha applied with 250 kg potash superphosphate/ha. They were grazed by 4-month-old red deer hinds (n=11/treatment) from mid-April 2000 until early March 2001. In mid-March 2001 (Year 2), the pastures were topdressed again as for Year 1, and the original hinds, now yearlings which had grazed as a single group between studies, were returned to their respective treatments in mid-April 2001 and remained on the trial until mid-March 2002. They were mated during April/May. The pastures were also grazed by pregnant mature hinds (n=8/treatment) from mid-May 2001. As the Cu status (i.e. liver Cu concentration) of the yearling hinds on the pasture treated with 6 kg CuSO4(.)5H2O/ha was not significantly different from the untreated animals, in late July 2001 the yearling and mature deer on this treatment were treated orally with 10 g CuO wire particles. The mature hinds calved in November and the yearling hinds in December. Pasture samples were collected at about monthly intervals to determine concentrations of Cu and other minerals. In Year 1, liver biopsies and blood samples were collected at 4-6-weekly intervals for determination of Cu concentrations. In Year 2, samples were collected similarly at 6-12-weekly intervals. Liver biopsies and blood were also collected from progeny, along with milk from their dams. Liveweights were determined at 3-7-monthly intervals, as well as data on calving/mortality rates. RESULTS: Pasture Cu concentrations before the application of CuSO4(.)5H2O were 6-9 mg Cu/kg dry matter (DM) and remained at this level in the untreated Control paddocks throughout the study. In Year 1, 28 days after treatment, pasture Cu concentration was 25 and 35 mg Cu/kg DM for the Low and High treatments, respectively; while at the same time for the same treatments in Year 2 it was 20 and 60 mg/kg DM, respectively. A second 60 mg Cu/kg DM peak also occurred on Day 85 in Year 2 with the High treatment. The pasture Cu concentration returned to 6-9 mg/kg DM, and there were no differences between treatments at Days 80 and 150 in Years 1 and 2, respectively. In Years 1 and 2, the Low treatment had no significant effect on the Cu status of the weaner and yearling hinds, respectively, when compared with that of animals grazing the untreated Control pastures. Weaner (Year 1) and yearling (Year 2) deer on the High treatment had significantly higher mean serum and liver Cu concentrations in the late winter and spring period when compared with those on untreated Control pastures. CuO wire particles increased the mean serum Cu concentration at Days 60 and 180, and liver Cu concentration at Day 60, in yearling hinds. A similar effect was observed in mature hinds. Regardless of Cu treatment, the liver Cu concentration of the 1-4-week-old progeny was markedly greater (p<0.001) than that of their dams, and then decreased significantly until weaning in March. In progeny of treated yearling hinds, but not mature hinds, serum and liver Cu concentrations were significantly higher (p=0.013) than progeny of untreated dams. CONCLUSION: Topdressing pastures with CuSO4(.)5H2O at a rate of 12 kg/ha, but not 6 kg/ha, in mid-March was effective in increasing the Cu status of weanling hinds; while pastures topdressed with 12 kg CuSO4(.)5H2O /ha in mid-March and dosing hinds with 10 g CuO in late July were effective in increasing the Cu status of pregnant hinds, and in the case of the yearling hinds, significantly improved the Cu status of their progeny from birth to weaning.     

Differences in copper and selenium metabolism between Angus (Bos taurus) and Brahman (Bos indicus) cattle

A 2-yr study was conducted at the Range Cattle Research and Education Center, University of Florida – Institute of Food and Agricultural Sciences (IFAS) (Ona, FL), to evaluate differences in the metabolism of Cu and Se of Angus (Bos taurus) and Brahman (Bos indicus) cattle. Thirty-two pregnant beef cows (n = 8 Brahman and 8 Angus/yr) were enrolled in the study in the first trimester of gestation. This study consisted of three phases: 1) restriction (day 0 to 90), 2) supplementation (day 91 to 150), and 3) calving. During all three phases, cows were individually fed and housed in partially covered drylot pens. During the restriction and supplementation phases, cows were provided a 1.5 kg/d of a grain-based concentrate supplement, which was fortified with flowers of S (50 g of supplemental S/cow daily; restriction phase) or Cu and Se (100 and 3 mg/d of Cu and Se, respectively; supplementation phase). Blood and liver samples were collected from all cows at 30 d intervals and from both cows and calves within 24 h of calving. Colostrum and milk samples were collected at calving and 7 d after birth. All data were analyzed using the MIXED procedure of SAS, where cow and calf were the experimental unit. During the restriction phase, a breed × day effect (P = 0.03) was observed where Brahman had greater liver Cu concentration than Angus cows in all sampling days. For liver Se concentration, a tendency (P = 0.07) for a breed effect was observed where Angus cows tended to have greater liver Se concentration than Brahman. During the supplementation phase, breed (P < 0.001) and day (P < 0.01) effects were observed, where Brahman cows had greater liver Cu concentration than Angus. For liver Se concentration, a day effect (P < 0.001) was observed, where liver Se concentration increased (P < 0.001) from day 90 to 120 and remained unchanged (P = 0.86) until day 150. At calving, no effects of breed (P = 0.34) were observed for liver Cu concentration of cows; however, Brahman calves tended (P = 0.09) to have greater liver Cu concentration than Angus calves. For Se liver concentration at calving, Angus cows tended (P = 0.07) to have greater liver Se concentration than Brahman cows; however, no breed differences (P = 0.70) were observed for liver Se concentration of calves at birth. In summary, substantial differences in multiple indicators of Cu and Se status were observed between Angus and Brahman cattle, implying that Angus and Brahman cattle possibly have different mechanisms to maintain adequate Cu and Se status.     

Use of 2,3,2-tetramine as a hepatic copper chelating agent for treatment of copper hepatotoxicosis in Bedlington terriers

Five Bedlington Terriers with inherited copper (Cu) hepatotoxicosis and with hepatic Cu concentrations ranging from 3,000 to 11,000 micrograms/g of dry weight (normal, less than 350 micrograms/g of dry weight) were treated daily for up to 200 days with 2,3,2-tetramine tetrahydrochloride. During treatment, no change was made in the dietary Cu intake, which ranged from 12 to 16 micrograms/g of dry diet. Concentrations of hepatic and serum Cu, iron, and zinc were determined before and at the conclusion of the treatment period. In one dog, 24-hour urinary Cu concentration was measured before and during treatment. A liver biopsy specimen obtained after treatment had significantly (P less than 0.05) reduced hepatic Cu concentration (3,282 micrograms/g of dry weight; a 54.9% reduction), compared with the pretreatment value (7,281 micrograms/g of dry weight). After treatment, there was an overall general lessening of the extent of hepatic morphologic damage. Cytochemical examination for Cu in rhodanine-stained biopsy specimens revealed decreased numbers of Cu-laden hepatic lysosomes. The mean daily urinary Cu concentration increased as much as 25-fold during 2,3,2-tetramine treatment. Hepatic iron and zinc concentrations and serum Cu concentrations remained within normal ranges after treatment. Clinical or laboratory evidence of 2,3,2-tetramine toxicosis was not detected during treatment. These findings indicated that in affected Bedlington Terriers, 2,3,2-tetramine was a safe and rapid chelating agent of hepatic Cu.     

Hepatic Lipidosis in Anorectic, Lactating Holstein Cattle: A Retrospective Study of Serum Biochemical Abnormalities

The association between hepatic lipidosis (HL) and disease in 59 anorectic, ketotic, lactating Holstein heifers and cows was investigated. Severe HL, as determined by histologic evaluation of liver tissue, was present in 46 animals; only half of these animals required intensive treatment for ketosis, and only half had serum biochemical evidence of liver disease, as determined by the presence of a test value of 2-fold or greater than the upper limit of the reference range for at least 2 of the 4 serum tests: gamma-glutamyl transferase, aspartate aminotransferase, and sorbitol dehydrogenase activities and bile acid concentrations. Most cattle with biochemical evidence of liver disease and severe HL had been lactating for 14 or more days. Cows that required intensive treatment inconsistently had serum biochemical evidence of liver disease.
Although cattle with severe HL had significantly higher serum bilirubin concentrations and aspartate aminotransferase and sorbitol dehydrogenase activities than cattle with less severe lipidosis, the specificity of abnormally high serum sorbitol dehydrogenase activity or bilirubin concentration for severe lipidosis was only 8%. Abnormally high serum aspartate aminotransferase activity was 83% sensitive and 62% specific for severe lipidosis. Serum glucose and total carbon dioxide concentrations were significantly lower in cattle with severe lipidosis than in those with mild or moderate lipidosis, and low serum glucose or total carbon dioxide concentrations were rare in cattle without severe lipidosis. From these data, we conclude that the use of a single biochemical or histopathologic criterion to define severity of disease or degree of liver compromise in anorectic, ketotic cows results in the misidentification of many animals.     

Glutathion peroxidase and glucose-6-phosphate dehydrogenase activities in bovine blood and liver

A total of 46 cattle, including 25 as control, 16 with glycogen degeneration and 5 with severe fatty degeneration were studied. Whole blood and liver tissue specimens were used to measure glutathione peroxidase (GSH-Px) and Glucose-6-Phosphate Dehydrogenase (G6PD) activities. The present study determined the value of these parameters in diagnosing glycogen and fatty degeneration in cattle from the point of the status of antioxidation and lipid peroxidation. The results showed a significant decrease in hepatic GSH-Px activity and a significant increase in hepatic G6PD activity in cases of fatty degeneration. On the other hand, there were no significant changes in erythrocytic and hepatic GSH-Px and G6PD activities in cases of glycogen degeneration. The results indicated lipoperoxidation process in the liver tissues increased in cases of fatty degeneration. Therefore, supplying animals suffering from fatty liver with sufficient quantities of nutrient antioxidants may be valuable when treatment is considered.     

Mineral concentrations of plasma and liver after injection with a trace mineral complex differ among Angus and Simmental cattle

To examine the effects of cattle breed on the clearance rate of an injectable mineral product, 10 Angus and 10 Simmental steers were blocked by breed and initial BW (332 ± 33 kg) and injected with either Multimin 90 (MM) or sterilized saline (CON) at a dose of 1 mL/45 kg BW. Multimin 90 contains 15 mg Cu/mL (as Cu disodium EDTA), 60 mg Zn/mL (as Zn disodium EDTA), 10 mg Mn/mL (as Mn disodium EDTA), and 5 mg Se/mL (as sodium selenite). Steers received a corn-silage-based diet, and inorganic sources of Cu, Zn, Mn, and Se were supplemented at NRC recommended amounts. Jugular blood was collected immediately before injection and at 8 and 10 h post-injection and on days 1, 8, and 15 post-injection. Liver biopsies were collected 3 d before injection and on days 1, 8, and 15 post-injection. Liver and plasma mineral concentration and glutathione peroxidase (GSH-Px) activity data were analyzed as repeated measures. Plasma concentrations of Zn, Mn, and Se were greater (P = 0.01) and Cu tended to be greater (P = 0.12) post-injection in MM steers compared with the CON steers. Regardless of treatment, Simmental cattle had lower plasma concentrations of Cu, Zn, and Se (P ≤ 0.05) when compared with Angus cattle. Erythrocyte GSH-Px activity was greater (P = 0.01) in MM steers compared with CON steers. Liver concentrations of Cu, Zn, and Se were greater (P = 0.05) in MM steers compared with CON steers post-injection. Liver Mn concentrations tended to be greater (P = 0.06) in MM steers compared with CON steers in the days post-injection. Interestingly, Simmental cattle exhibited greater (P = 0.01) liver Mn concentrations in the days after injection compared with Angus cattle (7.0 and 6.0 mg Mn/kg for Simmental and Angus cattle, respectively), regardless of treatment. It is unclear if this breed difference is biologically relevant; however, these data may suggest that differences in liver excretion of Mn exist between the two breeds. Overall, use of an injectable trace mineral increased liver concentrations of Cu and Se through the 15-d sampling period, suggesting that this injectable mineral is an adequate way to improve Cu and Se status of cattle through at least 15 d.     

Purification of a protein from serum of cattle with hepatic lipidosis, and identification of the protein as haptoglobin.

A protein that has 2 subunits with molecular weight of 35,000 and 23,000 was detected in serum of cattle with hepatic lipidosis (fatty liver). The protein was purified from serum obtained from a cow with fatty liver, and was identified as haptoglobin, which is known to have hemoglobin-binding capacity and to be an acute-phase protein. To assess the relevance of haptoglobin in fatty liver, cattle were classified in 3 groups (healthy control, haptoglobin-positive, and haptoglobin-negative); liver triglyceride content and several serum biochemical variables were evaluated for the 3 groups. Compared with the control and haptoglobin-negative cattle, haptoglobin-positive cattle had significantly (P less than 0.01) higher liver triglyceride content, serum bilirubin concentration, and aspartate transaminase activity. Serum haptoglobin concentration was high in slaughter cattle (27 of 40 cattle tested), particularly in cows (20/28).     

Echo-guided studies on portal and hepatic blood in cattle

The objectives of the present study were (1) to establish and maintain catheters non-surgically in the portal and hepatic veins of cattle for long-term monitoring of postprandially absorbed metabolites, and (2) to assess the safety of the technique used. Aspiration and catheterization were not performed if there was risk of bleeding, as determined by hemostatic data and enlargement of portal and hepatic veins. In five Holstein cattle, before and 60-min after feeding, percutaneous ultrasound-guided aspiration of portal and hepatic blood, together with jugular blood, was performed utilizing 17G x 17.5 cm needles. Concentration of ammonia nitrogen tended to be higher after feeding, but that of urea nitrogen and glucose were not significantly changed. In five other Holstein cows liver veins were catheterized with 14 x 16.5 cm guide-needles attached with 50 cm polyethylene tubes and blood samples were collected from portal, hepatic and jugular veins at 30-min intervals for 4.5 hr. The concentration of plasma acetate and 3-hydroxybutyrate (3-HB) were significantly increased after feeding, while that of lactate was not changed. Catheters in the portal and hepatic veins enabled the collection of blood samples up to 15 days (mean, 7 days). After a 7-day observation period, five cows were euthanised and necropsied. Changes in the intercostal musculature, peritoneum and liver were negligible. During the observation period, clinical profiles, aspartate aminotransferase, alanine aminotransferase and total and differential WBC counts were within normal ranges.     

The correlation between serum selenium and blood selenium in cattle.

The selenium (Se) concentration of paired blood and serum samples from cattle was determined by 2 methods: 1) atomic absorption spectroscopy using hydride generation (HG-AAS), and 2) inductively coupled argon plasma emission spectroscopy using hydride generation (ICP). Samples from 327 cattle were analyzed by HG-AAS, and samples from 344 cattle were analyzed by ICP. The data were examined by linear regression analysis, and the technique of inverse prediction was utilized to determine prediction intervals for estimating blood Se concentration from known serum Se concentration. The correlation coefficients, by simple linear regression of serum Se on blood Se, were 0.79 (r2 = 0.62) and 0.88 (r2 = 0.77) for the HG-AAS data and the ICP data, respectively. For the HG-AAS data, the inverse prediction formula for estimating blood Se when serum Se is known, at the 95% prediction interval, was [formula; see text]. For the ICP data, the inverse prediction formula for estimating blood Se when serum Se is known, at the 95% prediction interval, was [formula; see text]. The prediction intervals were quite wide, and the accuracy of estimating blood Se from a known serum Se was not useful for diagnostic purposes. The use of serum Se concentration to assess nutritional status of cattle with respect to Se does not appear to be appropriate.