An evaluation of the efficacy of injectable microencapsulated vitamin B12 in increasing and maintaining the serum and liver vitamin B12 concentrations of lambs

AIM: To develop a long-acting Vitamin B12 injection to prevent Co deficiency in sheep. METHODS: Formulations of microencapsulated Vitamin B12 in lactide-glycolide polymers were injected intramuscularly or subcutaneously into the anterior neck region of groups of 10 lambs and their efficacy determined from changes in serum and liver Vitamin B12 concentrations. RESULTS: The 95:5 lactide glycolide and the 100 lactide formulations containing more than 12.5% Vitamin B12 w/w significantly increased and maintained serum Vitamin B12 concentrations for at least 210 days as well as liver Vitamin B12 concentrations in treated lambs when compared with untreated controls. CONCLUSIONS: Injections of microencapsulated Vitamin B12 in lactide/glycolide copolymers are able to increase and maintain the Vitamin B12 status of lambs for at least 210 days. CLINICAL SIGNIFICANCE: Another option for the prevention of Co deficiency in sheep is now available using a long acting injectable Vitamin B12.     

The effect of increasing the Vitamin B12 status of Romney ewes on foetal liver Vitamin B12, milk Vitamin B12 and liver Vitamin B12 concentrations in suckling lambs

AIM: To determine the effect of increasing the Vitamin B12 status of the ewe on the Vitamin B12 supply to the suckling lamb. METHODS: The Vitamin B12 status of the ewe was increased during gestation and lactation by three injections of a long-acting preparation of Vitamin B12 microencapsulated in an organic acid polymer. The Vitamin B12 status of the ewes and suckling lambs was assessed from changes in serum and liver Vitamin B12 concentrations. RESULTS: Compared to untreated animals, serum and liver Vitamin B12 concentrations of the treated ewes were increased at least 70% during gestation. Foetal liver Vitamin B12 concentrations were increased 270%. Over the lactation, ewe serum and milk Vitamin B12 concentrations were increased at least 200% and 44%, respectively. The liver Vitamin B12 stores of the newborn lambs from Vitamin B12-treated ewes were depleted within 58 days. There were no significant differences in the serum Vitamin B12 concentrations of suckling lambs from Vitamin B12-treated and untreated ewes. CONCLUSION: Ewes with a high Vitamin B12 status will ensure an adequate supply of Vitamin B12 to their lambs for at least the first 30 days of life. CLINICAL SIGNIFICANCE: In flocks grazing Co-deficient pastures, treating ewes with a long-acting Vitamin B12 supplement at mating will prevent Vitamin B12 (Co) deficiency in ewes, as well as their lambs, until they can be treated at tailing at 4-6 weeks of age.     

Growth response to increasing doses of microencapsulated vitamin B12 and related changes in tissue vitamin B12 concentrations in cobalt-deficient lambs

AIM: To investigate growth response of cobalt deficient lambs to increasing doses of microencapsulated vitamin B12, and to measure associated changes in serum and liver vitamin B12 concentrations over 243 days. METHODS: From a flock grazing pastures that had low cobalt (Co) levels (about 0.06 mg Co/kg dry matter), 4-6-week-old lambs (n=137) were assigned to four groups and received either no treatment or a subcutaneous injection of 3.0, 4.5 or 6.0 mg of microencapsulated vitamin B12 on Day 1. At approximately monthly intervals, all lambs were weighed and blood samples were collected from a selection (n=10) of monitor animals, up to Day 243. Liver biopsies were also carried out on the monitor lambs (n=8) on Days 1, 124 and 215. RESULTS: The vitamin B12-treated lambs grew significantly faster (p<0.001) than untreated animals. Liveweights after 243 days were 28, 45, 45 and 47 kg for the untreated, 3.0, 4.5 and 6.0 mg vitamin B12-treated lambs, respectively. Of the initial group of untreated lambs, 68% had to be removed before the end of the trial because of substantial weight loss, but none of the treated animals were similarly afflicted. Serum vitamin B12 concentrations increased in all vitamin B12-treated lambs, reaching a peak at Day 25, and those of the 4.5 and 6.0 mg vitamin B12-treated lambs remained significantly higher (except at Day 124) than the untreated lambs to Day 187. However, at Day 124, but not Day 215, the liver vitamin B12 concentrations of treated lambs were two to three times higher than those of controls. CONCLUSIONS: The growth rates of Co deficient lambs were markedly improved by injection of 3.0, 4.5 or 6.0 mg of microencapsulated vitamin B12, and liveweights were maintained for at least 243 days. Serum vitamin B12 concentrations were related to this growth response; concentrations of <220 pmol vitamin B12/l were associated with a 95% probability that lambs were Co deficient and would thus respond to Co/vitamin B12 supplementation. Based on these data, the current New Zealand reference criteria for Co deficiency should be reviewed. CLINICAL SIGNIFICANCE: An injection of 3 mg microencapsulated vitamin B12 given to lambs at tailing will treat Co deficiency and will increase and maintain liveweights in a flock for up to 8 months.     

Effect of an injectable microencapsulated Vitamin B12 on serum and liver Vitamin B12 concentrations in calves

AIM: To evaluate the efficacy of a long-acting injectable microencapsulated formulation of Vitamin B12 in dairy calves. METHOD: Fifty calves, average liveweight 110kg, were randomly allocated to 5 groups of 10 animals and injected subcutaneously in the anterior neck with 0.12, 0.18, 0.24 and 0.3 mg Vitamin B12/kg liveweight using a formulation of microencapsulated Vitamin B12 in a lactide: glycolide copolymer. The untreated calves were injected with the same vehicle, without Vitamin B12. Subsequent changes in serum and liver Vitamin B12 concentrations were followed for 244 days. RESULTS: The microencapsulated Vitamin B12 significantly increased, then maintained serum and liver Vitamin B12 concentrations higher than those of untreated controls for at least 110 days. CONCLUSIONS: Injection of the microencapsulated Vitamin B12 at dose rates of 0.12 to 0.24 mg/kg liveweight will increase and maintain the Vitamin B12 status of calves for at least 110 days.     

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.     

Growth response to increasing doses of microencapsulated vitamin B12 and related changes in tissue vitamin B12 concentrations in cobalt-de cient lambs

AIM: To investigate growth responses of cobalt-deficient lambs to increasing doses of microencapsulated vitamin B₁₂, and to measure associated changes in serum and liver vitamin B₁₂ concentrations over 243 days.

METHODS: From a flock grazing pastures that had low cobalt (Co) levels (about 0.06 mg Co/kg dry matter), 4-6-week-old lambs (n=137) were assigned to four groups and received either no treatment or a subcutaneous injection of 3.0, 4.5 or 6.0 mg of microencapsulated vitamin B₁₂ on Day 1. At approximately monthly intervals, all lambs were weighed and blood samples were collected from a selection (n=10) of monitor animals, up to Day 243. Liver biopsies were also carried out on the monitor lambs (n=8) on Days 1, 124 and 215.

RESULTS: The vitamin B₁₂-treated lambs grew significantly faster (p<0.001) than untreated animals. Liveweights after 243 days were 28, 45, 45 and 47 kg for the untreated, 3.0, 4.5 and 6.0 mg vitamin B₁₂-treated lambs, respectively. Of the initial group of untreated lambs, 68% had to be removed before the end of the trial because of substantial weight loss, but none of the treated animals were similarly afflicted. Serum vitamin B₁₂ concentrations increased in all vitamin B₁₂-treated lambs, reaching a peak at Day 25, and those of the 4.5 and 6.0 mg vitamin B₁₂-treated lambs remained significantly higher (except at Day 124) than the untreated lambs to Day 187. However, at Day 124, but not Day 215, the liver vitamin B₁₂ concentrations of treated lambs were two to three times higher than those of controls.

CONCLUSIONS: The growth rates of Co-deficient lambs were markedly improved by injection of 3.0, 4.5 or 6.0 mg of microencapsulated vitamin B₁₂, and liveweights were maintained for at least 243 days. Serum vitamin B₁₂ concentrations were related to this growth response; concentrations of <220 pmol vitamin B₁₂/l were associated with a 95% probability that lambs were Co-deficient and would thus respond to Co/vitamin B₁₂ supplementation. Based on these data, the current New Zealand reference criteria for Co deficiency should be reviewed.

CLINICAL SIGNIFICANCE: An injection of 3 mg microencapsulated vitamin B₁₂2 given to lambs at tailing will treat Co deficiency and will increase and maintain liveweights in a flock for up to 8 months.     

Boluses of controlled release glass for supplementing ruminants with cobalt

Boluses of controlled release glass containing cobalt and weighing approximately either 60 g or 14.5 g were administered to 22 steers and 21 sheep respectively. The steers were housed and slaughtered at intervals between 17 and 145 days after dosing. The boluses released more than 0.85 mg cobalt daily. In both untreated and dosed animals serum and liver vitamin B12 concentrations were at the upper end of the normal range. Two types of glass were administered to sheep. In five wethers one glass released 0.07 mg cobalt per day, and in 16 grazing lambs a second glass released more than 0.15 mg cobalt per day. Fourteen of the boluses were recovered from the lambs up to 276 days after dosing. The concentration of B12 in serum of lambs increased significantly from a mean +/- sd of 1.64 +/- 0.47 to 2.02 +/- 0.04 ng/ml serum and the concentration in liver from 3.84 +/- 0.85 to 4.99 +/- 0.72 micrograms/g dry weight liver.     

Copper oxide needles administered during early pregnancy improves the copper status of ewes and their lambs

AIM: To determine the effect of copper oxide (CuO) needles administered orally to ewes in early pregnancy on the copper (Cu) status of ewes throughout gestation and lactation, and of their lambs from birth to weaning. METHODS: In mid-April, after mating, 12 twin-bearing ewes were given an oral capsule containing 5 g CuO needles while 12 others served as untreated controls. Changes in Cu status were monitored by determining serum and liver Cu concentrations on Days 1, 62, 117, 153, 185 and 216 in the ewes, and at 1, 36, 68 and 99 days of age for lambs. Pasture herbage samples were collected at about 60-day intervals for Cu, molybdenum (Mo), iron (Fe) and sulphur (S) determinations. RESULTS: Copper status of the ewe flock was adequate, as initial mean serum and liver Cu concentrations were 15 micromol/L and 1,060 micromol/kg fresh tissue, respectively. The CuO needles did not affect serum Cu concentrations of the ewes or their lambs. Mean serum Cu concentration of all lambs at birth was about half that of ewes (8 vs 17 micromol/L), regardless of Cu supplementation, and not until at least 68 days of age was it similar to the dams'. Liver Cu concentrations of lambs at birth were also lower than that of the ewes (380 vs 640 micromol/kg fresh tissue among the controls), but changed little over time. CuO treatment increased liver Cu concentration in ewes for at least 185 days and in lambs for 36 days (p<0.05). Among untreated ewes, there was a seasonal decline in mean liver Cu concentrations, which were highest in autumn and lowest in early spring (1,060 vs 370 micromol/kg fresh tissue). The mean pasture mineral concentrations were Cu 5.7, Mo 0.48, Fe 194 and S 2,900 mg/kg dry matter (DM). CONCLUSIONS: CuO needles administered to ewes in early pregnancy increased their Cu status through gestation and early lactation, and the Cu status of their lambs for 36 days from birth. Serum Cu concentration was not affected by treatment but a marked rise was observed in all lambs between birth and 10 weeks of age. CLINICAL RELEVANCE: Copper deficiency in young lambs may be conveniently and effectively prevented by treating ewes with CuO needles during early pregnancy. The serum Cu concentration in lambs <8 weeks old may not reflect the Cu status of the flock.     

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.     

Effect of colostrum treated by heat on immunological parameters in newborn lambs

Seventeen newborn lambs were fed with untreated colostrum (group A) and 18 lambs were fed with colostrum treated by heat (56 °C, 30 min) (group B). Blood samples were obtained at seven time points from day 0 to 38 of life. Clinical status and body weight were recorded and serum protein fractions and IgG levels were determined in blood. Phagocytosis of neutrophils from the lambs and opsonic capacity of their serum were examined using a flow cytometry method. As an indicator of in vivo cellular immunity, reactions to intradermal injection of phytohemagglutinin (PHA) were examined at 8 and 16 days of age. There were no clinical signs of disease and no significant differences in body weight between groups. Lambs from group B had lower total protein initially and lower γ-globulin and IgG levels until day 32 compared to lambs from group A (P < 0.01). Heat treatment of colostrum had no significant effect on serum opsonic capacity or phagocytosis by neutrophils and these functions increased with age. However, responses to PHA in lambs from group B were lower (P < 0.01) than group A at day 8 of life, but not at day 16, suggesting that cellular immunity was affected by the treatment of colostrum. These results showed that colostrum treated by heat impaired cellular and IgG levels in lambs, but not phagocytic function. However, these changes did not affect the health or performance of the lambs.     

Retention of selenium in tissues of calves, lambs, and pigs after parenteral injection of a selenium-vitamin E preparation.

Four each healthy weaned calves, lambs, and pigs raised in Indiana without selenium supplementation were killed, and their tissues were fluorometrically analyzed to establish base line selenium concentrations. The following mean selenium content (in ppm, wet weight) was found in calves, lambs, and pigs, respectively: liver, 0.12, 0.16, and 0.19; renal cortex, 0.63, 0.89, and 0.70; muscle, 0.05, 0.05, and 0.06. Eight each additional healthy weaned calves, lambs, and pigs were injected with a commercial selenium-vitamin E preparation at dose levels of 0.0825, 0.055, or 0.06 mg of Se (as selenite) per kilogram of body weight, respectively. Selenium content of tissues was measured in animals killed at 1, 7, 14, and 23 days after injection. In calves, concentrations in liver and kidney rapidly increased to moderate values and then slowly decreased, with mean concentrations after 23 days still somewhat greater than base line values. Concentrations for injection site tissue also rapidly increased to moderate values, but had decreased to base line values by 23 days after injection. In lambs, selenium content of liver was moderately increased after injection, but had decreased to base line values after 14 days; kidney and injection site did not have increased selenium content after injection. In pigs, liver and kidney had moderate initial increases in concentration of selenium, but these were at base line values after 14 days, and increase did not occur at injection sites.     

Effects of oral administration of levothyroxine sodium on serum concentrations of thyroid gland hormones and responses to injections of thyrotropin-releasing hormone in healthy adult mares

OBJECTIVE: To determine the effects of levothyroxine sodium (L-T4) on serum concentrations of thyroid gland hormones and responses to injections of thyrotropin-releasing hormone (TRH) in euthyroid horses. ANIMALS: 12 healthy adult mares. PROCEDURE: 8 horses received an incrementally increasing dosage of L-T4 (24, 48, 72, or 96 mg of L-T4/d) for weeks 1 to 8. Each dose was provided for 2 weeks. Four additional horses remained untreated. Serum concentrations of total triiodothyronine (tT3), total thyroxine (tT4), free T3 (fT3), free T4 (fT4), and thyroid-stimulating hormone (TSH) were measured in samples obtained at weeks 0, 2, 4, 6, and 8; 1.2 mg of TRH was then administered i.v., and serum concentrations of thyroid gland hormones were measured 2 and 4 hours after injection. Serum reverseT3 (rT3) concentration was also measured in the samples collected at weeks 0 and 8. RESULTS: Treated horses lost a significant amount of weight (median, 19 kg). Significant treatment-by-time effects were detected for serum tT3, tT4, fT3, fT4, and TSH concentrations, and serum tT4 concentrations were positively correlated (r, 0.95) with time (and therefore dosage) in treated horses. Mean +/- SD serum rT3 concentration significantly increased in treated horses (3.06 +/- 0.51 nmol/L for week 8 vs 0.74 +/- 0.22 nmol/L for week 0). Serum tT3, tT4, fT3, and TSH concentrations in response to TRH injections differed significantly between treated and untreated horses. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of levothyroxine sodium increased serum tT4 concentrations and blunted responses toTRH injection in healthy euthyroid horses.     

Serum and tissue concentrations of erythromycin in calves with induced pneumonic pasteurellosis

The effects of pneumonia on the pharmacokinetics of erythromycin administered IM and the tissue concentration changes with time were evaluated in 2-month-old calves. Pneumonia was induced by injection of Pasteurella haemolytica cultures through the thoracic wall into each lung. Six days prior to induction of pneumonia, erythromycin (15 mg/kg) was administered in a single IM dose. Erythromycin was administered again 48, 72, and 96 hours after injection of P haemolytica. On the third day of erythromycin administration (96 hours), the calves were serially euthanatized in groups of 4 calves each at 2, 5, 8, 12, 18, and 24 hours after the final dose was given. Tissue concentrations of erythromycin in kidney, liver, lung, muscle, CSF, and serum were determined. Neither the serum concentrations nor the overall pharmacokinetic values were significantly (P less than or equal to 0.05) changed by pneumonia. The concentrations of erythromycin were maximal at 5 hours for liver, muscle, and serum and at 8 hours for CSF, kidney, and lung. Serum and muscle concentrations were similar, whereas concentrations in CSF were lower than in serum and higher in kidney, liver, and lung. The lung/serum ratios were approximately 2.5 to 3 at 8 through 24 hours after IM administration. The peak concentration in lung was approximately 6 micrograms/g at 8 hours.     

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.     

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.     

Effect of intravenous infusion of recombinant ovine leptin on feed intake and serum concentrations of GH, LH, insulin, IGF-1, cortisol, and thyroxine in growing prepubertal ewe lambs

In sheep, serum concentrations of leptin change congruently with increases or decreases in nutritional status, while intracerebroventricular infusions of leptin dramatically suppress feed intake in well-fed lambs, and may also increase growth hormone (GH), and/or luteinizing hormone (LH) in undernourished lambs. The objective of the present study was to determine the effects of peripherally delivered ovine leptin, via intravenous infusions, on feed intake and serum concentrations of GH, LH, insulin, IGF-1, cortisol, and thyroxine. Twelve ewe lambs weighing 29.4 +/- 0.7 kg were infused intravenously with a linearly increasing dose of leptin or saline (n = 6 per group) for 10 days, reaching a maximum dose delivered of 0.5mg/h on day 10. Feed intake was assessed twice daily, and blood samples were collected every 10 min for 6 h on days 0, 2, 5, 8, and 10. Serum concentrations of leptin increased in leptin-treated lambs by day 2 (P = 0.05), and continued to increase to concentrations 9-fold greater than saline-infused lambs by day 10 (P < 0.001). Despite the substantial increase in serum leptin, feed intake did not differ between leptin and saline-infused lambs except on day 3.5 (P = 0.01). Furthermore, intravenous infusions of leptin did not significantly influence serum concentrations of insulin, cortisol, IGF-1, thyroxine, LH, or GH. Collectively, these observations contrast with the potent hypophagic effects of leptin when delivered intracerebroventricularly into well-fed lambs. The reasons for the disparate response of lambs treated intravenously with leptin, versus that reported for lambs treated intracerebroventricularly with leptin are not known, but may provide insight into the mechanism(s) of leptin resistance.     

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.     

Clinical, clinicopathologic, pathologic, and toxicologic alterations associated with gossypol toxicosis in feeder lambs

The toxicity of gossypol was studied in 20, 8-week-old feeder lambs that weighed approximately 16 kg. The lambs were allotted to 4 groups (A, B, C, D) and given (orally for 30 days) a daily dosage of gossypol (0, 45, 136, or 409 mg, respectively). Lambs were observed twice daily until they died or were euthanatized on the day the last dose was given. Clinical, electrocardiographic, clinicopathologic, pathologic, and toxicologic findings were recorded. All lambs given 409 mg of gossypol (group D) died before the end of the 30-day study. In this group, clinical signs included sudden death and/or chronic dyspnea syndromes. One group-B lamb had chronic dyspnea, but did not die. Electrocardiographic abnormalities observed in gossypol-treated lambs included increased amplitude of the T wave and decreased duration of the ST segment. Clinicopathologic alterations in group-D lambs included high serum total lactate dehydrogenase and lactate dehydrogenase liver-specific isoenzyme activities. Serum total creatine kinase activity decreased markedly in lambs of all groups treated with gossypol. Hemoglobinuria was detected in 2 group-D lambs. The average daily weight gain was remarkably consistent in lambs of all groups, except in 1 lamb each of groups D and B. Lambs were necropsied when they died or on day 30 of the study. The heart of gossypol-treated lambs weighted more than did that of untreated (control group A) lambs.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of disulfiram in experimentally induced vitamin E deficiency myopathy in lambs

Vitamin E deficiency myopathy (white muscle disease) was induced in 14 suckling lambs (2 experiments; 7 lambs/experiment) by addition of cod liver oil to the diet. Disulfiram, an antioxidant, was administered orally once each day to 8 of the 14 lambs at 2 different doses. Serum creatine kinase (CK) activity was measured weekly for 5 weeks. Increased CK activity was evident in some lambs beginning at week 3. By week 4, serum CK was abnormally increased in 5 of the 6 nontreated lambs (ie, disulfiram not given) and in 4 of the 8 treated lambs. The combined disulfiram groups had significantly lower serum CK values during the study (P less than 0.05). Serum alpha-tocopherol, measured on samples from week 5 for lambs of experiment 1, was significantly higher in treated lambs (P less than 0.01). Microscopic examination of the vastus lateralis muscle indicated that the most severe lesions, consistent with nutritional myopathy, were seen in nontreated lambs. Therefore, disulfiram may have an antioxidant effect in lambs with vitamin E deficiency.     

A review of tissue reference values used to assess the trace element status of farmed red deer (Cervus elaphus)

AIMS: This paper reviews the principles for the establishment of biochemical reference criteria for assessing the trace element status of farmed livestock and summarises data for copper, selenium, vitamin B12 and iodine for farmed red deer. COPPER: Enzootic ataxia and osteochondrosis occur when liver copper concentrations are below 60 micromol/kg fresh tissue, and serum copper concentrations are below 3-4 micromol/l. Growth responses to copper supplementation have been equivocal when blood copper concentrations were 3-4 micromol/l, but were significant when mean blood copper concentrations were 0.9-4.0 micromol/l. No antler growth or bodyweight response to copper supplementation was observed when blood ferroxidase levels averaged 10-23 IU/l (equivalent to serum copper concentrations of 6-13 micromol/l) and liver copper concentrations averaged 98 mumol/kg fresh tissue. These data suggest that 'deficient', 'marginal' and 'adequate' ranges for serum copper concentrations should be 5, 5-8, and 8 micromol/l, respectively, and those for liver copper concentrations should be 60, 60-100, and 100 micromol/kg, respectively. SELENIUM: White muscle disease has been reported in young deer with blood and liver selenium concentrations of 84-140 nmol/l and 240-500 nmol/kg fresh tissue, respectively. No growth-rate response to selenium supplementation occurred in rising 1-year-old deer when blood selenium concentrations were less than 130 nmol/l, the range in which a growth-rate response would be expected in sheep. VITAMIN B12: Vitamin B12 concentrations in deer are frequently below 185 pmol/l without clinical or subclinical effects. No growth response was observed in young deer with vitamin B12 concentrations as low as 75-83 pmol/l. A growth response to cobalt/vitamin B12 supplementation occurs in lambs with serum vitamin B12 concentrations 336 pmol/l. CONCLUSIONS: Data that can be used to establish reference ranges for assessing trace element status in deer are limited. More robust reference values for farmed red deer need to be established through further studies relating biochemical data to health and performance.