Use of vitamin B12 in joint lavage for determination of dilution factors of canine synovial fluid

OBJECTIVE: To test a modified saline (0.9% NaCl) solution joint washing (lavage) technique that includes the use of vitamin B12 as an internal marker for the evaluation of synovial fluid dilution in lavage samples from canine joints. SAMPLE POPULATION: 9 plasma samples obtained from blood samples of 9 healthy dogs and 9 synovial fluid samples aspirated from stifle joints of 9 cadaveric dogs. PROCEDURE: Photometric absorbances of 25% vitamin B12 solution, canine synovial fluid, and canine plasma were measured in a spectrophotometer to establish an optimal wavelength for analysis. Canine synovial fluid and plasma samples were mixed with the 25% vitamin B12 solution to obtain 1%, 3%, 5%, 10%, 20%, and 50% solutions of synovial fluid or plasma. Diluted synovial fluid and plasma samples were used to simulate joint lavage samples and to examine the possible interference of these substances (synovial fluid or plasma) with the absorbance of the 25% vitamin B12 solution in photometric analysis. RESULTS: The optimal wavelength was found to be at 550 nm. Canine synovial fluid and plasma samples did not interfere with the absorbance measurements of the 25% vitamin B12 solution up to a 50% dilution of plasma or synovial fluid. CONCLUSIONS AND CLINICAL RELEVANCE: The modified saline solution joint lavage method with the use of a 25% vitamin B12 solution as an internal standard provides an accurate and reliable technique for the evaluation of synovial fluid dilution in lavage samples from canine joints.     

Absence of a weight gain response to Vitamin B12 supplementation in weaned dairy heifers grazing pastures of marginal cobalt content

AIM: To obtain information on serum and liver vitamin B12 and urinary methylmalonic acid concentrations as diagnostic tests to predict a weight gain response to supplementation with vitamin B12 in young dairy cattle when grazing pasture of low cobalt content. Methodology. Forty dairy cattle (12 Friesian, 14 Friesian x Jersey and 14 Jersey) were allocated to two equal sized groups, treated and untreated, based on liveweight. At monthly intervals for 14 months, all animals were weighed, their serum and urine sampled, their liver biopsied and the pasture sampled from the paddocks they were grazing and going to graze. Serum and liver were assayed for vitamin B12 concentrations. For the first 5 months of the trial, urine was assayed for methylmalonic acid concentrations. Both washed and unwashed pasture samples were assayed for cobalt concentrations. RESULTS: No weight gain response occurred vitamin B12 supplementation in young growing cattle grazing pasture with a cobalt concentration of 0.04-0.06 mg/kg DM. For 5 months of the trial, liver vitamin B12 concentrations from untreated calves were in the range 75-220 nmol/kg and serum vitamin B12 concentrations were as low as 72 pmol/1. There was no associated growth response to supplementation. CONCLUSION: Further trials involving young cattle grazing pastures with cobalt concentrations less than 0.04 mg/kg DM are required to reliably determine liver and serum vitamin B12 concentrations at which growth responses to vitamin B12 or cobalt supplementation are likely under New Zealand pastoral grazing conditions.     

Measurement of vitamin B12 in the livers and sera of sheep and cattle and an investigation of factors influencing serum vitamin B12 levels in sheep

Modifications of a radioassay method for the analysis of vitamin B12 using chicken serum as the binder are described. This obviates the need to use individual serum blanks to correct for non-specific binding in vitamin B12 assays of the sera and livers of sheep and cattle. Samples with high vitamin B12 levels can be diluted prior to assay without loss of linearity. Recoveries of added cyanocobalamin or hydroxocobalamin were better than 95% and results correlated significantly with those obtained using a microbiological assay (Poteriochromonas malhamensis). Sera and liver samples stored for four weeks at temperatures ranging from -20 degrees to 22 degrees showed no change in vitamin B12 levels. Withholding food from sheep for 44 hours led to a marked increase in serum vitamin B12. This effect was also evident in sheep eating a limited amount of cut grass. In sheep at pasture there was no evidence of a diurnal variation in serum vitamin B12 levels. Serum vitamin B12 levels in sheep at pasture were shown to be an unreliable indicator of liver vitamin B12.     

Evaluation of a turbidimetric immunoassay for measurement of plasma IgG concentration in foals

OBJECTIVE: To validate a turbidimetric immunoassay (TIA) for measurement of plasma IgG concentrations in foals. ANIMALS: 36 foals. PROCEDURES: Blood samples were collected from foals before suckling and at 12 and 24 to 36 hours after birth. Plasma IgG concentrations were determined via a commercial single radial immunodiffusion (RID) assay. By use of goat anti-equine IgG antiserum and a spectrophotometer, a TIA was developed to measure plasma and serum IgG concentrations; the percentage light transmission was calibrated against RID assay-determined IgG concentrations. Assay repeatability and effects of serial dilution, sample type, and ambient temperature on assay results were evaluated. RESULTS: Serial dilution of plasma samples from foals 12 and 24 to 36 hours of age with presuckle plasma yielded percentage light transmission results that were highly inversely correlated (r = -0.95) with IgG concentrations determined via RID assay. Measurements of IgG in plasma and serum samples via TIA did not differ. When samples were assayed multiple times, the coefficient of variation was < 5.0%. Ambient temperature did not affect TIA results. At IgG concentrations of 400 and 800 mg/dL, TIA sensitivity was > 90%; specificity was 99.1% and 70.5%, respectively; and positive and negative predictive values were 98.1% and 71.5%, respectively, and 96.4% and 91.1%, respectively. CONCLUSION AND CLINICAL RELEVANCE: Plasma IgG concentrations in foals determined via the TIA and RID assay were highly correlated. The TIA rapidly yielded quantitative results and would be useful in clinical situations where intervention decisions are time dependent.     

Enzyme-linked immunosorbent assay for differentiation of the antibody response of cattle naturally infected with Brucella abortus or vaccinated with strain 19

Purified O chain of Brucella abortus was passively attached to polystyrene to differentiate antibody responses of cattle vaccinated with B abortus strain 19 from those of naturally infected cattle. In the indirect assay, using O polysaccharide as antigen, a single serum dilution was used and mouse monoclonal antibody to bovine L chain conjugated with horseradish peroxidase was the detection reagent. Measurable antibody was not found in sera of vaccinated cattle, except for 3 sera from cattle that were persistently infected with strain 19. Sera from 25 cattle infected with pathogenic strains contained antibody on the basis of results of indirect enzyme immunoassay, using smooth lipopolysaccharide or O chain as antigens, or results of competitive enzyme immunoassay, using the O-chain antigen. Results in sera from calves with experimentally induced Yersinia enterocolitica serotype 0:9 infection or inoculated with a low dose of B abortus strain 2308 were comparable with those in sera of cattle that were vaccinated with strain 19. The data correlated with those from competitive enzyme immunoassay, using one serum dilution and horseradish peroxidase-conjugated mouse monoclonal antibody to smooth lipopolysaccharide. On the basis of results of the indirect enzyme immunoassay, all sera (except those samples obtained before inoculation) contained antibody to smooth lipopolysaccharide.     

In vivo prediction of extracellular and intracellular water in cattle and sheep using thiocyanate and urea

Two experiments were conducted in cattle and sheep to determine the earliest time for thiocyanate equilibration with extracellular water. In Exp. 1, nine animals were infused to determine marker concentrations and sampling times. In Exp. 2, five steers were infused and then exsanguinated for tissue analyses. Thiocyanate equilibrated 22 to 31 min after infusion with a pool size equivalent to expected extracellular water. Plasma thiocyanate half-life averaged 29 h. Tissue concentrations 24 or 48 h after thiocyanate infusion were 20 to 24% of those observed in plasma for heart muscle and kidney and 6 to 8% in liver and skeletal muscle. A procedure is proposed for the in vivo estimation of empty body water (urea dilution), extracellular water (thiocyanate dilution) and, by difference, intracellular water in cattle and sheep, requiring only three blood samples, an initial sample and two samples taken 12 and 28 min after intravenous infusion of a urea-thiocyanate solution.     

A comparison of commercial kit methods for assay of vitamin b(12) in ruminant blood

Five commercially available radioisotope dilution (RID) kits for assay of Vitamin B(12) were obtained for evaluation with ovine and bovine serum; also, for comparison with our laboratory's (Lab 1) RID method and the methods of two independent laboratories, one using RID and the other a microbiological method. One commercial kit was found to be suitable for both species because negligible non-specific binding occurred when extraction was performed at 100 degrees C (212 degrees F). Of the other four kits, two were found to be suitable for ovine and bovine serum provided non-specific binding of the latter was measured. This was also the case with the Lab 1 method. Two of the commercial kits were found to be unsuitable for assay of Vitamin B(12)in ruminant blood.     

Changes in serum concentrations of methylmalonic acid and vitamin B12 in cobalt-supplemented ewes and their lambs on two cobalt-deficient properties

AIM: To determine concurrent changes in serum methylmalonic acid (MMA) and vitamin B12 concentrations of ewes and their lambs on cobalt-deficient properties, subsequent to cobalt supplementation. METHODS: Three experiments were carried out on two farms. Groups of ewes (n=25-50) were either supplemented with cobalt bullets during late pregnancy, 23-47 days before the mean lambing date, or left unsupplemented. In two experiments, lambs from within each group were supplemented directly by vitamin B12 injection at 3-weekly intervals from birth, and in the third experiment by injection with micro-encapsulated vitamin B12 at tailing and 3 months later. Pasture samples were obtained for analysis of cobalt content at each sampling time. Blood samples were obtained and liveweight recorded from ewes and lambs at approximately monthly intervals. On one farm (two experiments), liver and milk samples were obtained from ewes and liver samples from lambs. RESULTS: Serum vitamin B12 concentrations in unsupplemented ewes fell below 250 pmol/L during early lactation in all experiments and mean concentrations as low as 100 pmol/L were recorded. MMA concentration was maintained below 2 micromol/L in serum from supplemented ewes but increased to mean concentrations ranging from 7 to 14 micromol/L at the nadir of serum vitamin B12 concentration during peak lactation. A significant liveweight response to supplementation was recorded in ewes on one property, and the vitamin B12 concentration in the ewes' milk and in the livers of their lambs more than doubled. No liveweight-gain response to supplementation was observed in lambs on this property. Mean serum MMA concentrations in lambs ranged from <2 in supplemented, to 19.2 micromol/L in unsupplemented lambs, and the latter had concurrent serum vitamin B12 concentrations of >300 pmol/L. Pasture cobalt concentration was lowest at 0.04-0.09 microg/kg dry matter (DM) on the property on which responses in lambs occurred but considerably higher (>0.09 microg/kg DM) on the property on which responses in ewes occurred. On the second property, serum vitamin B12 concentrations in lambs at tailing were extremely low (100 pmol/L), irrespective of supplementation of dams with cobalt. Mean serum MMA concentration was increased to 20 and 42 micromol/L in lambs from supplemented and non-supplemented ewes, respectively. Weight-gain response to direct supplementation of lambs with vitamin B12 occurred during suckling in the latter, but not the former. Lambs from ewes supplemented with vitamin B12 showed a much bigger increase in serum vitamin B12 concentrations a month after supplementation than did lambs from unsupplemented ewes (+1,400 pmol/L vs + 650 pmol/L). CONCLUSIONS: Serum MMA concentration gave a more precise indication of responsiveness to vitamin B12 or cobalt supplementation than serum vitamin B12 concentrations in ewes and lambs. Neither very low serum vitamin B12 nor elevated MMA concentrations were necessarily indicative of responsiveness to supplementation in suckling lambs, but the latter gave an early indication of impending responsiveness. Supplementation of the ewe with a cobalt bullet appeared to protect the growth performance of the lamb for 90 days and influence the subsequent serum vitamin B12 response in the lamb to vitamin B12 supplementation. CLINICAL SIGNIFICANCE: Supplementing ewes with cobalt bullets in late pregnancy can improve the vitamin B12 status of their lambs, and modify their response to vitamin B12 supplementation.     

Concurrent changes in serum vitamin B12 and methylmalonic acid during cobalt or vitamin B12 supplementation of lambs while suckling and after weaning on properties in the South Island of New Zealand considered to be cobalt-deficient

AIM: To compare serum analyses of vitamin B12 and methylmalonic acid (MMA) as indices of cobalt/vitamin B12 deficiency in lambs around weaning. METHODS: Lambs on five properties, considered to be cobalt- deficient, were supplemented with either cobalt bullets, or short- or long-acting vitamin B12 preparations. Blood samples, and in some cases liver biopsies, and liveweights were obtained at monthly intervals. Serum samples were assayed for vitamin B12 and MMA and liver for vitamin B12 concentrations. Pasture cobalt concentrations were measured on three of the properties. RESULTS: Pasture cobalt concentrations were generally maintained below 0.07 microg/g dry matter (DM) on the properties sampled. Growth responses to supplementation were observed on only 2/5 properties, despite serum vitamin B12 concentrations being within the currently used 'marginal' reference range (336-499 pmol/L) for at least 3 months on all properties and in the deficient reference range (0-335 pmol/L) for at least 2 months on all farms except one. Serum MMA concentrations in supplemented lambs were <2 micromol/L, except in those animals sampled 1 month after receiving treatment with a short-acting vitamin B12 injection. Serum MMA concentrations in unsupplemented animals on properties on which no growth response to supplementation occurred generally reached peak levels of between 4 and 7 micromol/L at the nadir of serum vitamin B12 concentration. When a growth response was observed, differences in weight gain between supplemented and unsupplemented lambs occurred as mean serum MMA concentrations increased from 9 to 14 micromol/L. On one property where supplementation commenced before weaning, normal growth rates were maintained despite serum vitamin B12 concentrations of 140 pmol/L and serum MMA concentrations in excess of 40 micromol/L serum. CONCLUSIONS: The possibility that current serum vitamin B12 references ranges for diagnosis of cobalt deficiency are set too high and lead to over-diagnosis of responsiveness to cobalt/ vitamin B12 supplementation is discussed. The suggestion is made that serum MMA concentrations in excess of 9-14 micromol/L will provide a more reliable diagnostic test for cobalt deficiency. However, there was sufficient variation between properties in the relationships between cobalt concentrations of pasture and serum vitamin B12 or MMA concentrations to require more rigorous testing of the reliability of using serum MMA concentration for this purpose. The possibility that differences in rumen fermentation and therefore propionate and vitamin B12 production could be involved is discussed. The measurement of serum MMA and vitamin B12 appears to be of little value whilst the lamb is still suckling. CLINICAL SIGNIFICANCE: Serum MMA concentration may offer advantages over serum vitamin B12 concentrations in the diagnosis of a cobalt/vitamin B12 responsiveness in weaned lambs.     

Problems in the diagnosis and anticipation of trace element deficiencies in grazing livestock

Three principles governing the interpretation of biochemical criteria of trace element status are identified; they concern the relationships between the concentration of the marker and the intake of the element, the time on an adequate regimen and disturbances of tissue function. From these principles, the concentrations of liver copper, liver vitamin B12 and ovine serum vitamin B12 are shown to be insensitive indices of deficiency but good indices of surfeit. Plasma copper less than 9 mumol/litre is a good index of marginally deficiency but values may have to fall below 3 mumol/litre before there is risk of dysfunction and loss of production in sheep and cattle. Serum vitamin B12 values below 188 pmol/litre are indicative of functional deficiency in sheep whereas cattle with values between 38 and 76 pmol/litre may be only marginally deficient. Concentrations of methylmalonic acid in the plasma greater than 5 mumol/litre may offer a surer guide to diagnosis of functional vitamin B12 deficiency. Blood selenium or glutathione peroxidase concentrations may be unreliable in diagnosing selenium-responsive conditions because other nutrients determine what is adequate. For all elements the surest diagnosis is an improvement in growth or health in response to a specific supplement. The adoption of preventive measures should be prompted by biochemical evidence of marginal deficiencies in animals (rather than soils or pastures) although economic responses will not necessarily follow.     

Validation of a chemiluminescent enzyme immunometric assay for plasma adrenocorticotropic hormone in the dog

Background: The concentration of canine adrenocorticotropic hormone (ACTH) is usually determined by radioimmunoassay. However, chemiluminescent assay techniques have many advantages for clinical endocrine testing. Objectives: The objectives of this study were to validate a commercially available chemiluminescent assay for determination of canine ACTH concentration and to determine whether protease inhibitors are appropriate for use in the chemiluminescent assay system. Methods: Biological specificity was evaluated by treatment of 3 dogs with ovine corticotropin‐releasing hormone (CRH) followed by serial measurements of ACTH and by comparison with a previously validated immunoradiometric assay. All samples were collected both in the presence and absence of aprotinin, a protease inhibitor. The assay was further evaluated by measurement of intra‐assay precision, interassay precision, and recovery after dilution. Results: Baseline ACTH concentrations ranged from 5.6 to 15.3 pg/mL, and maximum ACTH concentrations of 158 to 1240 pg/mL were observed 30–60 minutes after CRH administration. Plasma samples collected with aprotinin had significantly lower ACTH concentrations than did samples collected without aprotinin. The intra‐assay coefficients of variance (CVs) ranged from 4.1 to 8.2%, and interassay CVs ranged from 4.6 to 14.8%. Recovery after dilution with canine plasma ranged from 93.4 to 103.0% of predicted concentration; however, inadequate recovery was observed with other diluents. There was a high correlation with the immunoradiometric assay (r= .925) but a significant negative bias (‐32.9, 95% confidence interval ?50.8 to ?14.9). Conclusions: This chemiluminescent assay is a valid technique for measurement of ACTH in canine plasma. ACTH concentration measured by chemiluminescence is lower than that measured by immunoradiometry. Aprotinin decreases the measured concentration of ACTH, and this effect should be taken into account when interpreting results. Diluents supplied with the kit should not be used for dilution of canine samples.     

The comparison of thiobarbituric acid reactive substances (TBARS) concentrations in plasma and serum from dairy cattle

The aim of this study was to compare thiobarbituric acid reactive substances (TBARS) concentrations in serum, plasma with heparin (heparin plasma), and plasma with ethylenediaminetetraacetic acid disodium salt (EDTA plasma) as anticoagulants from dairy cattle. Serum, heparin plasma, and EDTA plasma TBARS were not sufficiently strongly correlated to allow accurate prediction of one set of values from the other. Heparin plasma TBARS concentrations were found to be lower, and were affected by the duration of mixing during the assay process. The results suggest that it is necessary to differentiate TBARS concentrations between different sample types such as serum, heparin plasma, and EDTA plasma. For measurements of TBARS concentrations in cattle, EDTA plasma samples may be more suitable than the other samples.     

Interrelationships among liposoluble vitamins in ruminants

Plasma concentrations of vitamins A and E were examined in sheep, and a transitory decrease was observed after a single massive dose of vitamin D3 (5 X 10(6) IU) was administered orally or parenterally. Administration of a large dose of vitamin E to sheep decreased plasma retinol concentrations within 72 hours, but thereafter, the plasma retinol concentrations returned to near baseline values. Oral administration of a single pharmacologic dose of dl-alpha-tocopherol (5 g) to sheep caused a slow increase of this vitamin in the blood plasma. In cattle, a single IM administration of 3 liposoluble vitamins (A, D3, and E) at acceptable concentrations had no detectable influence on plasma alpha-tocopherol concentrations with the sampling intervals used. Plasma concentrations of alpha-tocopherol in these cattle showed a marked seasonal pattern; the concentrations increased from January to a peak in July, with a subsequent decrease in the fall. Also reported are estimates of inter- and intraindividual variation in plasma liposoluble vitamin concentrations.     

Measurement of feline intact parathyroid hormone: Assay validation and sample handling studies

A two-site intact immunoradiometric assay was validated for the measurement of parathyroid hormone (PTH) in the cat, by assessment of precision, sensitivity and specificity. The mean intra- and interassay coefficients of variation were 12·7 and 12·8 per cent, respectively. The sensitivity of the assay was 3·90 pgl/ml. The presence of carboxyl fragment interference was suspected from the non-parallel dilution of two of seven feline samples and the decreased recovery of PTH on addition of carboxyl terminal fragment PTH to samples. The plasma PTH concentration, measured using this assay, in 40 clinically healthy cats was 10·9 ± 5·3 pg/ml (mean ± SD). The assay demonstrated appropriate responses in cases of secondary renal hyperparathyroidism, hypercalcaemia of malignancy and iatrogenic hypoparathyroidism. Storage characteristics would permit posting of samples. This assay will therefore provide a useful non-invasive tool for the diagnosis of disorders of calcium metabolism in the cat.     

Effects of concentrations of cyanocobalamin in the gestation diet on some criteria of vitamin B12 metabolism in first-parity sows

In swine nutrition, little is known about the role of vitamin B(12) in the reproductive processes. The current study was undertaken to obtain information on the dose-response pattern of different metabolic criteria related to the homeostasis of vitamin B(12) and homocysteine in gestating sows receiving various concentrations of dietary vitamin B(12) (cyanocobalamin). Homocysteine is a detrimental intermediate metabolite of the vitamin B(12)-dependent remethylation pathway of Met. Forty nulliparous (Large White x Landrace) sows were randomly assigned during gestation to dietary treatments containing 5 concentrations of cyanocobalamin (0, 20, 100, 200, or 400 microg/kg). During lactation, a diet containing 25 microg of cyanocobalamin/kg (as-fed) was given to all sows. During gestation, plasma vitamin B(12) increased as concentrations of dietary cyanocobalamin increased (linear and quadratic, P < 0.01) and the effect persisted during lactation (21 d postpartum) both in plasma (linear and quadratic, P < 0.05) and the liver (linear and quadratic, P < 0.04). Plasma homocysteine decreased with concentrations of cyanocobalamin provided to sows during gestation (linear, quadratic, and cubic, P < 0.01). At parturition, vitamin B(12) in colostrum increased as concentrations of cyanocobalamin increased (linear and quadratic, P < 0.01), but the treatment effect persisted (linear, P = 0.01) only up to 1 d postfarrowing. However, in piglets there was no treatment effect (P = 0.59) on plasma vitamin B(12) before colostrum intake, but a linear effect of concentrations of cyanocobalamin (P = 0.04) was observed 1 d later. Plasma homocysteine in piglets during lactation decreased with increasing concentrations of cyanocobalamin given to sows in gestation (linear and quadratic, P < 0.01). Based on a broken-line regression model, the concentrations of dietary cyanocobalamin that maximized plasma vitamin B(12) and minimized plasma homocysteine of sows during gestation were estimated to be 164 and 93 microg/kg, respectively. The maximal residual responses in sows and piglets during lactation were observed with treatments of 100 or 200 microg of cyanocobalamin/kg. The dietary cyanocobalamin concentration necessary to optimize the response of these metabolic criteria remains to be refined within lower and narrower ranges of cyanocobalamin concentrations (i.e., <200 mg/kg). Moreover, the biological significance of such concentrations of cyanocobalamin needs to be validated with performance criteria by using greater numbers of animals during several parities.     

Dose response effects of long-acting injectable vitamin B12 plus selenium (Se) on the vitamin B12 and Se status of ewes and their lambs

AIM: To determine the effect of increasing doses of long-acting injectable vitamin B12 plus selenium (Se) given pre-mating on the vitamin B12 and Se status of ewes and their lambs from birth to weaning. METHODS: Four groups of 24 Poll Dorset ewes each were injected 4 weeks pre-mating with different doses of a long-acting vitamin B12 + Se product, containing 3 mg vitamin B12 and 12 mg Se per ml. The treatment groups received 5 ml (15 mg vitamin B12 + 60 mg Se), 4 ml (12 mg vitamin B12 + 48 mg Se), 3 ml (9 mg vitamin B12 + 36 mg Se), or no vitamin B12 or Se (control). Twelve of the twin-bearing ewes per group were selected for the study. Efficacy of the product was evaluated from changes in the concentrations of vitamin B12 in serum and liver, and of Se in blood, liver and milk in the ewes during gestation and lactation, and in their lambs from birth to weaning. Pasture samples in paddocks grazed by the ewes and lambs were collected at about 2-monthly intervals from 200-m transects. RESULTS: The flock was Se-deficient, as the mean initial concentration of Se in the blood of ewes was 182 (SE 20.3) nmol/L. Compared with untreated controls, all doses significantly (p < 0.01) increased concentrations of Se in the blood of ewes for at least 300 days. Selenium concentrations in milk were likewise increased throughout lactation, as were those in the blood and liver of lambs. The mean concentration of vitamin B12 in the serum of ewes was initially > 1,000 pmol/L, but this decreased within 28 days to < 460 pmol/L. Treatment with the 5-ml and 4-ml doses raised serum vitamin B12 concentrations of ewes for at least 176 days (p < 0.01), while their lambs had significantly greater concentrations of vitamin B12 in serum and liver for less than 37 days after birth. Tissue concentrations and duration of elevation of both vitamin B12 and Se were proportional to the dose administered. The mean concentrations of Se and cobalt (Co) in the pastures were 32 and 74 microg/kg dry matter (DM), respectively. CONCLUSIONS: Injecting ewes from a Se-deficient flock 4 weeks prior to mating with 48 or 60 mg Se and 12 or 15 mg vitamin B12 increased and maintained the Se status of ewes for at least 300 days, and of their lambs from birth to weaning. The vitamin B12 status of ewes was increased for at least 176 days and that of their lambs for less than 37 days. Due to the proportional nature of the response to increasing dosage, the dose rate of the formulation tested can be adjusted according to the severity of Se and Co deficiency in a flock. CLINICAL SIGNIFICANCE: A single subcutaneous injection of vitamin B12 + Se administered pre-mating to Se-deficient flocks is likely to prevent Se deficiency in ewes and their lambs until weaning, as well as increase the vitamin B12 status of ewes and their lambs until 5 weeks after lambing.     

Sex-related responses to vitamin B12 and trace element supplementation in prime lambs

Bodyweight, plasma vitamin B12 and blood selenium concentrations were monitored in prime lambs given different forms of supplementation at 2 sites in separate years. At the first site treatment groups comprised control, vitamin B12 injection, selenium given orally and a combination of vitamin B12 and selenium. At the second site cobalt and selenium supplied in a glass bullet was compared with an untreated group. Significant sex-related responses were observed to treatment in terms of bodyweight and at site 2 in plasma vitamin B12 concentrations. A marked bodyweight response to glass bullet supplementation was observed in castrated male lambs but not in ewe lambs. These studies show that sex differences should be considered when investigating trace element deficiencies.     

Absence of a weight gain response to Vitamin B12 supplementation in weaned dairy heifers grazing pastures of marginal cobalt content

Aim. To obtain information on serum and liver vitamin B₁₂ and urinary methylmalonic acid concentrations as diagnostic tests to predict a weight gain response to supplementation with vitamin B₁₂ in young dairy cattle when grazing pasture of low cobalt content.

Methodology. Forty dairy cattle (12 Friesian, 14 Friesian × Jersey and 14 Jersey) were allocated to two equal sized groups, treated and untreated, based on liveweight. At monthly intervals for 14 months, all animals were weighed, their serum and urine sampled, their liver biopsied and the pasture sampled from the paddocks they were grazing and going to graze. Serum and liver were assayed for Vitamin B₁₂ concentrations. For the first 5 months of the trial, urine was assayed for methylmalonic acid concentrations. Both washed and unwashed pasture samples were assayed for cobalt concentrations.

Results. No weight gain response occurred to Vitamin B₁₂ supplementation in young growing cattle grazing pasture with a cobalt concentration of 0.04-0.06 mg/kg DM. For 5 months of the trial, liver Vitamin B₁₂ concentrations from untreated calves were in the range 75-220 nmol/kg and serum vitamin B₁₂ concentrations were as low as 72 pmol/1. There was no associated growth response to supplementation.

Conclusion. Further trials involving young cattle grazing pastures with cobalt concentrations less than 0.04 mg/kg DM are required to reliably determine liver and serum Vitamin B₁₂ concentrations at which growth responses to Vitamin B₁₂ or cobalt supplementation are likely under New Zealand pastoral grazing conditions.     

Effects of biological type of beef steers on vitamin D, calcium, and phosphorus status

Feedlot steers (n = 36) from three biological types (Bos indicus, Bos taurus-Continental, and Bos taurus-English) were used to determine the Ca, P, and vitamin D3 status of feedlot cattle. The USDA yield and quality grade traits were measured at slaughter, and the concentrations of vitamin D3 (VITD) and the metabolites 25-hydroxyvitamin D3 (25-OH D) and 1,25-dihydroxyvitamin D3 (1,25-(OH)2 D) were determined in LM, liver, kidney, and plasma. Plasma and muscle Ca and P concentrations also were determined. Biological type of cattle affected a number of carcass traits. Carcasses from Bos taurus-English cattle had more marbling, resulting in higher quality grades (P < 0.05). Carcasses from Bos taurus-Continental cattle had lower calculated yield grades (P < 0.05) than did carcasses from cattle in the other biological types. In general, differences in carcass traits resulting from biological type were consistent with other reports. Plasma and LM Ca and P concentrations were not affected (P = 0.06) by biological type of cattle, indicating that Ca and P homeostasis is a conserved trait across the different types of cattle. Plasma VITD and 25-OH D concentrations were not affected (P = 0.41) by biological type, whereas plasma 1,25-(OH)2 D concentration was lower (P < 0.05) in Bos taurus-English cattle than in Bos taurus-Continental and Bos indicus cattle. Liver VITD and 25-OH D were not affected by biological type (P = 0.76), but liver 1,25-(OH)2 D concentration was greater (P < 0.05) in Bos indicus cattle than in Bos taurus-Continental cattle. Kidney vitamin D metabolite concentrations were not affected by biological type of cattle (P = 0.21). Muscle VITD concentration was greater (P < 0.05) in Bos taurus-English cattle than in the other two biological types, and muscle 25-OH D concentrations were greater (P < 0.05) in Bos taurus-English cattle than in Bos indicus cattle. Muscle 1,25-(OH)2 D concentration was less (P < 0.05) in the Bos taurus-Continental cattle than in the other two biological types. Cooking eliminated vitamin D metabolite differences among the biological types. Our results suggest that Bos indicus cattle had greater 1,25-(OH)2 D (the biologically active form) in tissues, and greater 1,25-(OH)2 D plasma concentrations than Bos taurus cattle. Thus, the need for VITD supplementation and optimal levels of Ca and P in feedlot diets might differ between Bos indicus and Bos taurus cattle.