A comparison of vitamin B12 levels in the liver and serum of sheep receiving treatments used to correct cobalt deficiency

The efficacies of four methods, used for the prophylaxis of cobalt deficiency in sheep as measured by the elevation of liver and serum vitamin B₁₂ levels, were compared in marginally deficient sheep over 14 weeks. The methods used were weekly drenches of either cobalt sulphate or cobalt chelate (EDTA); three-weekly injections of hydroxocobalamin, and ruminal cobalt pellets.

On the basis of elevated liver and serum vitamin B₁₂ levels, chelated cobalt was shown to be available to rumen microflora for the synthesis of vitamin B12. However, at no stage were liver and serum vitamin B₁₂ levels of sheep receiving the chelate significantly different from those receiving the same amount of cobalt as the sulphate.

After five, three-weekly injections of hydroxocabalamin liver vitamin B₁₂ levels were significantly higher (p < 0.01) than for the other treatments, with the exception of cobalt sulphate.

Cobalt pellets led to an initial rapid and significant rise in serum vitamin B₁₂ when compared with the other treatments. However, at four weeks there was no significant difference between treatment groups for serum vitamin B₁₂. Fourteen weeks after the administration of cobalt pellets, serum and liver. vitamin B₁₂ levels in this group were not significantly different from those of untreated sheep. At this time, three out of 12 sheep had lost their pellets.     

Effect of ingestion of soil on the iodine, copper, cobalt (vitamin B12) and selenium status of grazing sheep

AIM: To determine the impact of ingestion of soil on the iodine (I), selenium (Se), copper (Cu) and cobalt (Co; vitamin B12) status of young sheep. METHODS: Twenty young sheep were divided into two groups; one group was fed lucerne pellets, while the other group was fed lucerne pellets plus 100 g soil, for 63 days. At the end of the study the animals were blood-sampled, slaughtered, and the liver removed, and concentrations of I, Cu, vitamin B12 and Se were determined. RESULTS: The ingestion of soil significantly increased concentrations of I and vitamin B12 in serum, but had no effect on concentrations of Cu and Se in serum/blood and liver, and vitamin B12 in liver. CONCLUSION: Ingested soil can be a significant source of I and Co (vitamin B12) for grazing sheep.     

Trace element supplementation of sheep: evaluation of various copper supplements and a soluble glass bullet containing copper, cobalt and selenium

The effect of 5 different copper supplements on copper status of Merino sheep at pasture was examined. Transient increases in plasma copper concentrations occurred following treatment with copper calcium EDTA, copper sulphate, and glass bullets impregnated with copper, cobalt and selenium, but these increases were not considered to be a reliable indication of changes in liver copper reserves. Sheep receiving the glass bullet or 2.5 g oxidised copper wire particles orally or 50 mg copper as copper calcium EDTA parenterally had liver copper concentrations significantly greater than those of untreated sheep for periods up to 51 weeks. Oral doses of 30 mg copper as copper oxychloride, or 300 mg copper as copper sulphate did not alter the liver copper reserves at any stage. The bodyweight and greasy fleece weights were not significantly altered by any of the copper supplements. Plasma vitamin B12 concentrations in sheep given the glass bullet were increased for about 5 months. Thereafter increased plasma vitamin B12 concentrations were observed in all sheep. Blood selenium concentrations were also high during the experiment thereby preventing the evaluation of the glass bullet as a source of selenium.     

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.     

The occurrence of Anthrax in New Zealand

A field outbreak of facial eczema occurred during a vitamin B12 response trial in young growing sheep. Pasture cobalt levels were in the low range for sheep <0.08 mg/kg, 1.358 µmol/kg) and mean (of 3) liver vitamin B12 levels in the sheep were low (>100 nmol/kg) during the period in which facial eczema occurred.

Mean serum vitamin B12 levels of the untreated group were low (>185 pmol/1) for the two months (January and February) preceding the period of facial eczema. However, levels showed an approximate 3.5 fold increase in both cobalt supplemented and unsupplemented groups with the onset of facial eczema in March. From February to March the mean serum vitamin B12 and glutamyl transferase (GGT) activity showed parallel increases with a positive correlation (r = 0.73) between log serum vitamin B12 and log serum GGT activity during the period January to July for both groups. This finding suggested that the increase in serum vitamin B12 was due to sporidesmin induced liver damage.

The diagnostic implication is that, in areas where facial eczema is a problem, liver is the sample of choice for determining vitamin B12 status, because sporidesmin toxicity can elevate low serum vitamin B12 levels to diagnostically normal levels.     

The effect of sporidesmin toxicity on ovine serum vitamin B12 levels

A field outbreak of facial eczema occurred during a vitamin B12 response trial in young growing sheep. Pasture cobalt levels were in the low range for sheep (<0.08 mg/kg, 1.358 micromol/kg) and mean (of 3) liver vitamin B12 levels in the sheep were low (<400 nmol/kg) during the period in which facial eczema occurred. Mean serum vitamin B12 levels of the untreated group were low (<485 pmol/l) for the two months (January and February) preceding the period of facial eczema. However, levels showed an approximate 3.5 fold increase in both cobalt supplemented and unsupplemented groups with the onset of facial eczema in March. From February to March the mean serum vitamin B12 and glutamyl transferase (GGT) activity showed parallel increases with a positive correlation (r = 0.73) between log serum vitamin B12 and log serum GGT activity during the period January to July for both groups. This finding suggested that the increase in serum vitamin B12 was due to sporidesmin induced liver damage. The diagnostic implication is that, in areas where facial eczema is a problem, liver is the sample of choice for determining vitamin B12 status. because sporidesmin toxicity can elevate low serum vitamin B12 levels to diagnostically normal levels.     

WHITE LIVER DISEASE OF SHEEP

SUMMARY Outbreaks of ovine white liver disease (WLD) on 7 farms in eastern Victoria were investigated. Most occurred in late spring and mainly affected lambs 3 to 6 months old, with a morbidity of 20 to 100% and mortality of 8 to 15%. Clinically affected lambs showed illthrift, emaciation and bilateral, serous, ocular discharge. Clinical pathology showed mild anaemia, elevated serum liver enzymes (GGT, OCT, AST) and low levels of serum vitamin B12. Grossly, the livers were pale, fatty and friable; microscopically there was parenchymal fatty change, bile duct proliferation and ceroid pigmentation. Liver cobalt values were consistently low (mean 0.4 ± 0.4 μmol/kg D.W.). Levels of cobalt in pasture from 2 properties were very low (0.34 μmol/kg D.W.). The diagnosis of white liver disease was made on the basis of clinical features, specific liver pathology and low cobalt status. Treatment trials established that cobalt injections or oral bullet administration resulted in clinical improvement, significant weight gains, and improved serum vitamin B12 levels. WLD did not recur in previously affected sheep using these treatments. However, when blocks containing cobalt were available continuously, WLD recurred 2 years after the initial outbreak.     

Vitamin B12 responses to cobalt pellets in beef cows

Objective To assess the effectiveness of cobalt pellets in maintaining adequate vitamin B₁₂ in beef cows on pasture of low cobalt content.
Design A field experiment in a herd grazing cobalt deficient pasture.
Animals Mature Murray Grey cows.
Procedure Cows were given a single oral dose of 0, 1, 2 or 4 cobalt pellets (30 g pellets containing 30% by weight cobaltic oxide) with a selenium pellet and a grub screw. Samples of blood, liver, faeces and milk for chemical analyses were collected at intervals over a period of 2 years after treatment.
Results A single cobalt pellet raised liver vitamin B₁₂ concentration of cows above that of untreated cows for at least 28 weeks, and 2 or 4 pellets for 57 weeks. Plasma vitamin B₁₂ concentration was an unreliable indicator of the effectiveness of cobalt pellet therapy. Milk vitamin B₁₂ and faecal cobalt concentrations increased in response to cobalt pellet therapy.
Conclusion These studies show that one cobalt pellet will prevent vitamin B₁₂ inadequacy in beef cows for between 28 and 57 weeks; two or four pellets will prevent inadequacy for 57 to 75 weeks. Milk vitamin B₁₂ concentration may be a useful indicator of the effectiveness of cobalt pellets in increasing the vitamin B₁₂ supply in lactating cows.     

Effect of cobalt deficiency in the pregnant ewe on reproductive performance and lamb viability

Reproductive performance and lamb viability in cobalt sufficient and subclinically deficient ewes, and from ewes experiencing repletion from and depletion into cobalt deficiency, were investigated in two experiments. The sheep were fed a cobalt deficient ration and supplementation was by oral dose according to treatment. The treatments had a significant effect (P less than 0.001) on ewe serum vitamin B12 and methylmalonic acid concentrations. There were no significant effects on ewe liveweight, condition score or conception rate. Cobalt deficient ewes produced fewer lambs and had more stillbirths and neonatal mortalities than cobalt sufficient controls. Lambs from deficient ewes were slower to start suckling (P less than 0.05), had reduced concentrations of serum immunoglobulin G and zinc sulphate turbidity levels (P less than 0.05), and had lower serum vitamin B12 and higher methylmalonic acid concentrations (P less than 0.05), than lambs from cobalt sufficient dams. Cobalt supplementation in either the first or second half of pregnancy only did not fully alleviate these adverse effects.     

Duration of effective benefit from administration of graded oral doses of cobalt to sheep

The rapidity and duration of the response of sheep to graded oral doses of cobalt, given as the sulphate or as a cobalt-supplemented anthelmintic Panacur SC, was assessed by the determination of serum vitamin B12 and methylmalonic acid concentrations. The rapidity of the response to the treatments, which ranged from 1 to 250 mg cobalt, was unaffected by dose rate but the effective duration was dose related. There was some evidence of a cumulative effect from repeat treatments of the anthelmintic at three weekly intervals. It is suggested that the optimum inclusion rate for cobalt in anthelmintics lies within the range 21 to 100 mg/treatment.     

Caprine hepatic lipidosis induced through the intake of low levels of dietary cobalt

Forty-one, 10-week-old newly weaned goats were randomly allocated into two groups, namely control (n=22) and treated (n=19). Kids in both groups were fed Rhodegrass hay ad libitum that contained < 0.1 mg/kg DM cobalt and 150 g/day of a commercially prepared ruminant concentrate that contained approximately 0.12 mg/kg DM cobalt. This diet provided the minimum daily requirement of cobalt as specified for sheep. The treated goats were supplemented with bi-monthly subcutaneous injections of 2000 microg of hydroxycobalamin. All goats were weighed and blood samples collected monthly for haematological, clinical biochemical and serum vitamin B12 analysis. After a 10-month experimental period the goats were slaughtered. The control animals exhibited significantly (P<0.05) lower weight gains, and had dry scruffy hair coats. In addition, there was a decline in erythrocyte counts, mean haemoglobin, packed cell volume, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration. Controls also exhibited significantly (P<0.05) lower levels of total serum proteins and elevated levels of serum alkaline phosphatase compared to treated goats. Fourteen (63.6%) of the control goats developed pathology consistent with reported field cases of hepatic lipidosis associated with low liver levels of cobalt. Only one (5.3%) of the treated goats developed hepatic lipidosis. Contrary to previous reports that suggested that goats are less sensitive to low levels of dietary cobalt than sheep, it is apparent that this is not the case with Omani goats. This is the first report of the induction of hepatic lipidosis in goats due to feeding low levels of cobalt in their diet.     

A comparison of vitamin B12 levels in the livers and sera of sheep measured by microbiological and radioassay methods

Five radioassay methods (commercial kits) were evaluated for the determination of vitamin B₁₂ in sheep serum. They provided results that correlated significantly (P<0.01) with those obtained using a microbiological assay.

One radioassay method was evaluated for analysing vitamin B₁₂ in liver, and the results were in excellent agreement with those obtained by the microbiological method.

Little correlation between serum and liver vitamin B₁₂ levels was found. The limited diagnostic value of serum vitamin B₁₂ concentrations for detecting cobalt deficiency in sheep is discussed.     

Investigation into diagnosis and treatment of cobalt deficiency in lambs.

The development of cobalt deficiency was studied in 30 Scottish Blackface lambs grazing pasture on a soil containing 0-17 ppm cobalt. By the end of an eight-week period 50 per cent of lambs were subjectively appraised as showing signs of cobalt deficiency. After a further period lasting four weeks, during which three groups of lambs were studied (one group, least affected by cobalt deficiency, acted as control, the second received a single cobalt bullet and the third received oral doses of 200 mg cobalt chloride at the beginning of the period and three weeks later). Mean urinary formiminoglutamic acid (FIGLU) concentrations were inversely related to serum vitamin B12 concentrations and increased from 0-08 to 0-20 mumole per ml in group 1, and decreased to virtually zero within one week of treatment in groups 2 and 3. The use of serum vitamin B12 and urinary FIGLU concentrations in the diagnosis of cobalt deficiency in sheep are discussed.     

The retention and efficacy of soluble-glass boluses for providing selenium, cobalt and copper to sheep

The efficacy and retention of prototype and commercial (Cosecure) soluble-glass boluses containing selenium, cobalt and copper and having a similar size, density and composition were evaluated in separate 12-month trials with sheep grazing low selenium pastures but with adequate cobalt and copper levels. In both trials, sheep confirmed by X-ray as containing a bolus grew at a significantly greater rate than control sheep. This was attributed to the correction of a selenium deficiency. Although liver copper, and liver and serum vitamin B12 levels were significantly greater in treated sheep than controls on some occasions, the differences were not as great or consistent as with blood selenium levels. The rate of bolus loss was high and in both trials, approximately 40% of the sheep had lost the bolus after 6 months. No prototype boluses were present after 12 months. Intact Cosecure boluses were recovered from 3 sheep out of 30 after 12 months whilst one animal contained a part bolus. The recovered boluses were approximately 45% lighter than when originally administered. Bolus loss did not appear to be due to complete dissolution. In sheep which had lost the bolus, blood selenium levels fell with a half-life of 43 +/- 10 days.     

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.     

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.     

Reference curves to diagnose cobalt deficiency in sheep using liver and serum vitamin B12 levels

Reference curves demonstrating the relationship between serum or liver vitamin B12 and weight gain were derived from the examination of 16 published and 48 unpublished N.Z. trials. From these curves probability of obtaining an economic reponse (>10g/day body weight increase) for any serum or liver vitamin B12 can be determined. No significant (P<0.05) weight gain responses occurred to vitamin B12 or cobalt treatment in trials with mean serum vitamin B12 levels above 500 pmol/l or liver vitamin B12 levels greater than 500 nmol/kg. The reference curves were therefore derived from trials with vitamin B12 levels below these levels; 36 trials with serum vitamin B12 and 19 trials with liver vitamin B12 data. The mean vitamin B12 level at the mid point of the weight gain response period was selected from each trial. Examination of serum vitamin B12 reference curves for spring, summer, autumn and winter indicated that curves derived from data closest to the middle of January (summer) adequately reflected response to treatment at any time during the first year of life. Reference curves for liver vitamin B12 also used data closest to middle of January. This was partly because insufficient liver data was available to compare seasonal variations. The fitted response curve approached 0 gram/day at 500 pmol/l for serum vitamin B12 and 375 nmol/kg for liver vitamin B12. The minimum vitamin B12 level at which an economic response to treatment (>10 g/day) is not likely was 336 pmol/l for serum and 282 nmol/kg for liver.     

Effectiveness of α-tocopherol and selenium supplements in preventing lupinosis-associated myopathy in sheep

Objective To compare the effectiveness of combined selenium and α-tocopherol acetate treatments in preventing lupinosis-associated myopathy in sheep.
Design Measurement of plasma muscle and liver enzymes, and histopathological examination of muscle and liver in the treatment groups.
Procedure The treatments were: subcutaneous injections of selenomethionine and vitamin E (sc(SeM+E)) , an intraruminal selenium pellet and oral doses of vitamin E and intramuscular injections of selenomethionine combined with either oral doses of vitamin E (imSeM+orE) or intramuscular injections of vitamin E in an oily carrier. Another group received no supplements, while a control group was given selenium pellets on day 0 and fortnightly oral doses of vitamin E from day 0 to 72. To produce lupinosis-associated myopathy, the sheep were fed a diet low in vitamin E and given repeated injections of a crude extract of Diaphorthe toxica. Groups sc(SeM+E) and imSeM+orE were stressed by dosing with protected polyunsaturated fatty acids from day 56 onwards.
Results Lupinosis-associated myopathy was induced in all unsupplemented sheep. In these sheep the storage of Se increased and that of vitamin E decreased. The subcutaneous treatment was highly effective in preventing lupinosis-associated myopathy and also produced the highest vitamin E concentrations in plasma and liver. Supplemental vitamin E was more efficacious than supplemental Se. Concentrations of vitamin E in the livers of sheep given intramuscular vitamin E were higher than expected based on plasma concentrations. Oral doses of vitamin E proved the least effective method of increasing concentrations in liver. Lupinosis did not affect Se concentrations in liver or muscle.
Conclusion The sc(SeM+E) treatment is highly effective in preventing lupinosis-associated myopathy but needs to be further assessed when selenium and vitamin E are both limiting in the diet.     

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.     

Reduction of the prevalence of footrot on New Zealand farms by vaccination

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 μg/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 μmol/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 μmol/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 μmol/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 μmol/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 μmol/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.