Aqueous central cavity in aspartate transcarbamylase from Escherichia coli.

A three-dimensional x-ray diffraction study of aspartate transcarbamylase to 5.5-angstrom resolution, with the aid of four isomorphous heavy atom derivatives, indicates the presence of a central aqueous cavity approximating an oblate spheroid about 25 by 50 by 50 angstroms in dimension, within a molecule about 90 by 110 by 110 angstroms in largest dimensions.

In vitro metabolism of selenite in sheep blood: factors controlling the distribution of selenium and the labelling of plasma protein.

Some factors controlling the distribution of Na275SeO3 in sheep blood were studied in vitro. After centrifuging Na275SeO3-incubated blood most of the radioactivity was found in the plasma. The labelling of plasma protein by 75Se was dependent on the presence of erythrocytes. The degree of labelling of plasma protein increased with erythrocyte concentration. When phosphate-buffered saline-washed erythrocytes were suspended in phosphate-buffered saline and incubated with Na275SeO3 the majority of the 75Se was detected in the erythrocytes. On incubating these labelled erythrocytes with unlabelled plasma there was a transfer of radioactivity to the plasma. The calculated activation energy for the labelling of plasma was 107.52 kJ/mol. Albumin was shown not to be a principal acceptor of 75Se from the erythrocytes by ammonium sulphate precipitation of radioactive plasma. Addition of Na2SeO3 to the labelled blood resulted in the transfer of 75Se from plasma to the erythrocytes. Radioactive plasma incubated at 37 degrees C was thermolabile with respect to its 75Se content whereas in whole blood the degree of 75Se binding to plasma protein did not vary suggesting that a recycling of selenium was occurring in blood. From the results presented an in vitro model of selenium metabolism in blood is postulated.

An electrophoretic analysis of sheep plasma protein labeled with Na2 75SeO3 in vivo.

Following an intravenous injection of 75Se, sodium selenite plasma samples were analyzed by two-dimensional electrophoresis. 75Se was detected by indirect autoradiography. From 0.5 to 53 hr postinjection of 75Se, 21 75Se peptides were detected. Both the isoelectric points and molecular weights of these peptides are reported. The molecular weights of the peptides ranged from 20,000 to 70,000 daltons.

75Selenium-labeled sheep plasma: the time course of changes in 75selenium distribution.

Sheep fed rations containing 0.1 ppm selenium were labeled by intravenous injection of radioactive sodium selenite or selenocystine. Gel filtration of serially collected plasma samples indicated that, with time, there was a transition from mercaptan sensitive to high mol wt mercaptan and protein solubilizer resistant selenoproteins. Radiolabeled plasma samples collected from selenite and selenocystine labeled sheep were dialyzed against buffer containing 2-mercaptoethanol or protein solubilizer. No difference in the stability between selenite- and selenocystine-labeled plasma could be detected.

Investigations of the use of clay minerals and prussian blue in reducing the transfer of dietary radiocaesium to milk.

Two experiments were performed with lactating dairy cattle to assess the efficacy of clay minerals and Prussian Blue (AFCF form) in controlling the transfer of dietary radiocaesium to milk. In Experiment 1, bentonite was included in the diet at 0, 300, 600 and 900 g d-1 and the transfer of radiocaesium from silage to milk was determined. Bentonite inclusion significantly (P less than 0.001) depressed the transfer of radiocaesium to milk with no benefit in increasing the dietary inclusion above 600 g d-1 when a 73% reduction was observed. In Experiment 2, the effectiveness of bentonite (300 g d-1), clinoptilolite (300 g d-1) and Prussian Blue (3 g d-1) as dietary additives was compared. All treatments significantly (P less than 0.001) depressed the transfer of dietary radiocaesium to milk. Clinoptilolite was less effective than bentonite and both treatments were considerably less effective than Prussian Blue, the reductions being 35%, 62% and 85% respectively.

Studies on the uptake, partition and retention of ionic and fallout radiocaesium by suckling and weaned lambs.

In the first part of this study 9 one week old lambs were fed, ad libitum, a liquid milk replacer diet containing 950 Bq 137Cs kg-1, for a period of 21 days. Absorption of the 137Cs from the milk replacer was over 90% during this period. The lambs were weaned onto a silage diet and then allocated to 2 groups (A and B). Group A was fed silage contaminated with fallout radiocaesium and group B with the same silage additionally contaminated with ionic 134CsCl. After a 3 week period the diets offered were changed over between the groups. During the silage feeding period the uptake of added ionic 134Cs was approximately twice that of fallout 134Cs present in the silage. These studies highlight the difficulties in using studies with the ionic form of the isotope to predict the animals' response to fallout contaminated feed.

Studies of the transfer of dietary radiocaesium from silage to milk in dairy cows.

Four experiments were carried out to examine the transfer of radiocaesium from silage to milk by dairy cows. Three experiments assessed the effects of stage of lactation, yield and the feeding of forage only on this transfer. It was found that maximum milk radiocaesium concentrations were achieved rapidly (within 5 days) after the start of feeding contaminated silage. The transfer coefficients (Fm) were low ranging from 0.002 to 0.003 d l-1. The fourth experiment examined the effects of alternately feeding silages, of relatively high and low radiocaesium contents, on a 2 d cycle on the milk radiocaesium content. It was observed that the milk radiocaesium content changed rapidly in response to the amount of radiocaesium ingested.

The effects of prussian blue provided by indwelling rumen boli on the tissue retention of dietary radiocaesium by sheep.

A study was carried out to examine the effects of a form of Prussian Blue, provided by indwelling rumen boli, on the tissue retention of dietary radiocaesium by sheep. A single bolus providing 20-24 mg PB d-1 achieved a 42% reduction in tissue radiocaesium content. Multiple boli were not successful due to their loss from the rumen. The preparation used did not affect the health and well being of the animals.

Use of erythrocyte copper:zinc superoxide dismutase activity and hair or fleece copper concentrations in the diagnosis of hypocuprosis in ruminants.

Changes in the activity of superoxide dismutase, a copper-containing enzyme in erythrocytes (ESOD), and changes in copper in hair or fleece were compared with the changes in plasma copper during copper depletion and repletion in steers, lambs and ewes. During depletion the decline in ESOD began later than the decline in plasma copper: the lag varied from 0 to 80 days in individual steers and it was least evident when young rapidly growing lambs were subjected to severe depletion. ESOD activity eventually declined at only one-third to one-seventh of the rate shown by plasma copper, the difference being particularly marked in severely depleted lambs. Repletion of ESOD began after repletion in plasma copper and continued more slowly particularly in growing animals. Hair and fleece copper also responded relatively slowly to depletion and repletion. Low values of ESOD and hair or fleece copper may have diagnostic significance, indicating a more prolonged or intense deficiency of copper and a higher probability of clinical and production responses to copper therapy than low plasma or liver copper values.

The levels of selenium and glutathione peroxidase activity in blood of sheep, cows and pigs.

The levels of selenium (Se) and glutathione peroxidase (GPX) in the blood of sheep, cows and pigs under farm conditions were examined. Sheep appear to form two distinct groups, namely high Se and GPX and low Se and GPX. The high group gave ranges of 133-249 ng/ml and 77-179 iu/g Hb for blood Se and GPX respectively, while the low group showed levels of 21-67 ng/ml and 2-20 iu/g Hb. Overall sheep blood showed a high correlation between Se and GPX (r = 0-92, P less than 0-001). Cow bloods formed one group, all having low Se and GPX levels except for a single outlier. Omitting this animal, the overall ranges were 9-72 ng/ml and 6-36 iu/g Hb for Se and GPX respectively. Blood Se and GPX activity were significantly correlated (r = 0-59, P less than 0-001). Pigs formed a single group also, with the difference that while their blood Se was high, the corresponding blood GPX activities were relatively low. Overall ranges were 93-193 ng/ml and 17-69 iu/g Hb for Se and GPX respectively. Correlation between blood Se level and GPX activity in this species was not significant (r = 0-27, P more than 0-1).

The effect of the duration and type of stress on some serum enzyme levels in pigs.

The effects of blood sampling from the ear vein, jugular vein and collection on exsanguination, on serum CK, LDH, GOT and PK activities were investigated. The activities of LDH and PK were higher in ear vein vena cava samples. The effect of pre-slaughter handling was also investigated. In general there was a significant progressive increase in serum enzyme activities from farm to slaughter, although the magnitude of the change varied for different enzymes.

Variation in the activities of glutathione peroxidase and superoxide dismutase and in the concentration of copper in the blood in various breed crosses of sheep.

Activities of the enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were examined in the blood of 364 ewes of a lowland grazing flock comprising Scottish Blackface ewes and crosses of that breed with Border Leicester, Cambridge, East Friesland, Oldenburg and Texel rams. Half were bled at 16 months old and again two years later, and half at 28 months old. Variation attributable to breed of sire was not statistically significant (P less than 0.1) for GSH-Px, but was significant for whole blood copper concentration (P less than 0.01) and for SOD activity (P less than 0.05) although the latter was complicated by a large breed by age interaction involving two of the breeds. The correlation of SOD activity (per g haemoglobin) and whole blood copper concentration was 0.5, and the intraclass correlation between the estimates of SOD activities two years apart was 0.7.

Experimental 3-nitro-4-hydroxyphenylarsonic acid toxicosis in pigs.

Five times the recommended dose of 3-nitro-4-hydroxyphenylarsonic acid (187.5 mg kg-1) induces a pathognomonic clinical syndrome in pigs. The main clinical features are not continually present but are inducible only by exercise. From day 11 on the experimental diet a nervous syndrome was inducible. This manifested as muscle tremors and clonic convulsive episodes. Paraparesis developed by day 22 and paraplegia by day 33. Liver arsenic levels plateaued at 5.4 +/- 1.3 mg kg-1. The experiment confirms field observations that 3-nitro produces a characteristic toxicological syndrome, which is distinct from that of arsanilic acid. It also confirms that 3-nitro has a higher absolute toxicity than arsanilic acid in pigs as well as a lower margin of safety.

Contagious equine metritis--use of gas liquid chromatography in identifying the causal agent.

Cellular fatty acid compositions of contagious equine metritis isolates and three reference Haemophilus equigenitalis cultures were determined by gas chromatography. The chromatographic data were standardised and normalised fatty acid methyl ester (FAME) profiles were produced. The profiles were compared visually and similarity indices were determined using a computer peak matching method. There was little difference between the profiles of the three reference strains, each strain being characterised by three major fatty acids; C 18:1, C 16:0 and 30H-C 14:0. Variations in cultural conditions had no significant effect on the FAME profiles. The identification of laboratory isolates using the technique was in agreement with the presumptive identification based on the currently recommended tests and an improvement on the confirmatory serological identification. The FAME profiles provided confirmation of identity where it was not possible to use the presently recommended serological procedures. The authors recommend the gas chromatography technique for use in the diagnostic laboratory as an adjunct to the presently accepted identification methods.

The incorporation of Na2 75SeO3 into sheep erythrocytes.

Injection of Na2 75SeO3 into untreated, phlebotomized, and phenylhydrazine treated sheet has shown that the rate of 75SeO3-incorporation into erythrocytes is dependent on the degree of stimulation. Analysis of labeled erythrocytes by gel filtration and two-dimensional electrophoresis has indicated that the transient labeling of a hemoglobin-like peptide is the only protein labeled in addition to glutathione peroxidase.

Sequential changes in plasma methylmalonic acid and vitamin B12 in sheep eating cobalt-deficient grass.

Methylmalonic acid (MMA) concentrations are elevated in plasma as a result of vitamin B12 deficiency. This study reports the sequential changes in plasma MMA in lambs maintained on a cobalt-deficient pasture compared with supplemented controls. The results indicate that MMA is elevated in the early stages of deficiency, preceding the onset of loss of production and clinical signs of disease. It remains elevated as long as the lambs are unsupplemented with cobalt (Co). The most striking clinical sign was a loss of body condition as opposed to weight. The defect in the methylmalonyl CoA mutase is obviously an early defect in cobalt deficiency.

Use of sodium hydroxide treated selenium deficient barley to induce vitamin E and selenium deficiency in yearling cattle.

Selenium deficient barley grown in Northern Ireland was treated with sodium hydroxide to deplete it of vitamin E. Housed cattle fed a complete diet based on this treated barley developed nutritional degenerative myopathy, showing that spontaneous myopathy in yearling cattle can be the result of vitamin E and selenium deficiency alone. The diet used is as effective and cheaper than others presently in use for inducing degenerative myopathy.

Lack of effect of selenium supplementation on the incidence of weak calves in dairy herds.

A double blind controlled trial was carried out on four commercial dairy herds, to investigate the effect of one 50 mg injection of selenium, 10 days before parturition, on the incidence of weak calves. Although the treatment marginally increased the selenium status of treated calves it did not decrease the incidence of the weak calf syndrome.

Milk fever controls: comparison of 1-alpha and vitamin D3 in conjunction with induced parturition.

The efficacies of vitamin D3 and its 1-alpha hydroxyl derivative (1-alpha) in controlling clinical milk fever, hypocalcaemia and hypophosphataemia in parturient cows have been compared. A corticosteroid was used in some cases to optimise and control the interval between prophylactic treatment and parturition. Our observations suggest that the combination of 1-alpha and corticosteroid was particularly valuable and could be used in the development of a successful prophylactic regime. This conclusion is supported by both clinical and biochemical measurements.

Nitrogen dioxide (silo gas) poisoning in pigs.

Toxic silo gases are a potential danger to livestock housed in close proximity to silos. On the fourth day of ensiling, five fattening pigs were found dead in a pen adjoining a grass silo. Post mortem examination revealed extensive lung damage and methaemoglobinaemia. A dense reddish-brown gas was concentrated at floor level to a height of 1 m in the pen and had diffused into adjoining pens, where dry and suckling sows and litters were showing signs of respiratory distress and weakness. The gas was identified as a mixture of nitrogen dioxide and dinitrogen tetroxide. These gases may be produced in the early stages of silage making. In this case, they had accumulated in a slurry channel below the silo and leaked through an adjoining wall into the piggery. The production and toxicological effects of silo gases are discussed.