Serum cobalamin concentrations in healthy cats and cats with non-alimentary tract illness in Australia

Objective   To determine a reference range for serum cobalamin concentration in healthy cats in Australia using a chemiluminescent enzyme immunoassay and to prospectively investigate the prevalence of hypocobalaminaemia in cats with non-alimentary tract disease.
Design   Prospective study measuring serum cobalamin concentrations in clinically healthy cats and cats with non-alimentary tract illness.
Procedure   Blood was collected from 50 clinically healthy cats that were owned by staff and associates of Veterinary Specialist Services or were owned animals presented to Creek Road Cat Clinic for routine vaccination. Blood was collected from 47 cats with non-alimentary tract illness presented at either clinic. Serum cobalamin concentration was determined for each group using a chemiluminescent enzyme immunoassay.
Results   A reference range for Australian cats calculated using the central 95th percentile in the 50 clinically healthy cats was 345 to 3668 pg/mL. Median serum cobalamin concentration in 47 cats with non-alimentary tract illness (1186 pg/mL; range 117–3480) was not significantly different to the median serum cobalamin of the 50 healthy cats (1213 pg/mL, range 311–3688). Using the calculated reference range one sick cat with non-alimentary tract illness had a markedly low serum cobalamin concentration.
Conclusion   Although hypocobalaminaemia is uncommon in sick cats with non-alimentary tract illness in Australia, its occurrence in this study warrants further investigation.     

Degenerative myelopathy associated with cobalamin deficiency in a cat

A severe myelopathy was observed in a 9-year-old neutered male cat with a clinical history of chronic pancreatitis associated with deficiency of serum cobalamin and folates concentrations, and progressive spinal ataxia. The spinal cord lesions mainly involved the dorsal columns of the caudal cervical and cervico-thoracic segments, and were characterized by diffuse vacuolated myelin sheaths and axonal degeneration, marked gliosis, fibrosis and presence of gitter cells. The pancreas showed severe atrophy of the exocrine tissue, periductular fibrosis and infiltration of inflammatory cells, consistent with chronic interstitial pancreatitis. This condition can be accountable for cobalamin deficiency, as the pancreas is the only source of intrinsic factor in cats. The spinal cord lesions in the cat of this report resembled the subacute combined degeneration of the spinal cord described in human beings with cobalamin deficiency and hence a similar pathogenetic mechanism is hypothesized.     

Hypocobalaminaemia is uncommon in cats in the United Kingdom

Recent work has highlighted the importance of cobalamin deficiency in cats with a range of alimentary tract diseases. The primary aim of our study was to determine the incidence of subnormal cobalamin concentrations in sick cats with and without alimentary system disorders. Firstly, serum cobalamin concentrations were measured in a population of cats, with and without gastrointestinal (GI) disease, evaluated at a referral hospital. In the second part of the study, the incidence of cobalamin deficiency was assessed in samples submitted to a commercial laboratory specifically for cobalamin measurement. For both studies, a validated radioimmunoassay was used to measure serum cobalamin concentrations (reference range: > 150 pg/ml). In the first part of the study, 132 cats were included and none of these cats had subnormal cobalamin concentrations (median=1,172; range: 278 to >2,000). There were no differences in cobalamin concentrations between cats with alimentary system disorders, and those with diseases of other organs. In the second part, 682 samples were submitted for cobalamin assay over a period of 3 years, and only one cat had a result below the reference range (median=794; range: 147 to >2,000). Cobalamin deficiency was rare in the population tested and this may suggest that the incidence of this biochemical abnormality is less common than reported in the USA.     

Organic acidemia in a young cat associated with cobalamin deficiency

Objective: To describe a case of severe metabolic acidosis and encephalopathy secondary to cobalamin (Cbl) deficiency in a young cat. Case summary: A 4‐year‐old spayed female domestic short hair cat weighing 2.5 kg, presented with a 2‐day history of neurological signs referable to the cerebrum. The cat was evaluated for similar episodes twice before, however, no definitive diagnosis was made for either visit. On presentation the cat was minimally responsive and had a metabolic acidosis and ketonuria with no apparent reason, such as lactic acidosis or diabetic ketoacidosis. The patient was diagnosed postmortem with an organic acidemia secondary to low Cbl levels. New or unique information provided: The purpose of this report is to alert emergency clinicians to be suspicious of an organic acidemia in any case of metabolic acidosis and ketonuria that cannot be explained. Early identification of inherited organic acidemias in domestic animals may allow prompt and appropriate treatment of these conditions.     

Suspected acquired hypocobalaminaemic encephalopathy in a cat: resolution of encephalopathic signs and MRI lesions subsequent to cobalamin supplementation

PRESENTING SIGNS AND INITIAL INVESTIGATIONS: An 8-year-old female spayed British shorthair cat was presented with a history of waxing and waning neurological signs. Neuroanatomical localisation was consistent with a diffuse forebrain disease. Blood ammonia concentration was increased. Abdominal ultrasonography and a bile acid stimulation test were normal. Magnetic resonance imaging (MRI) revealed hyperintense, bilaterally symmetrical, diffuse lesions on T2-weighted sequences, predominantly, but not exclusively, affecting the grey matter. Serum cobalamin (vitamin B12) concentration was low. Hypocobalaminaemia resulting in a urea cycle abnormality was considered a likely cause of the hyperammonaemia. TREATMENT: Daily cobalamin injections resulted in a rapid clinical improvement. Eight weeks into treatment neurological examination was unremarkable and there was complete resolution of the MRI lesions. CLINICAL IMPORTANCE: This is the first reported case of acquired feline hypocobalaminaemia resulting in an encephalopathy. Additionally, this case is unique in describing reversible brain MRI abnormalities in a cobalamin-deficient companion animal.     

Evaluation of cats fed vegetarian diets and attitudes of their caregivers

OBJECTIVE: To determine motivation and feeding practices of people who feed their cats vegetarian diets as well as taurine and cobalamin status of cats consuming vegetarian diets. DESIGN: Cross-sectional study. ANIMALS: 34 cats that had been exclusively fed a commercial or homemade vegetarian diet and 52 cats that had been fed a conventional diet for > or = 1 year. PROCEDURES: Participants were recruited through a Web site and from attendees of a national animal welfare conference. Caregivers of cats in both groups answered a telephone questionnaire regarding feeding practices for their cats. Blood was obtained from a subset of cats that had been fed vegetarian diets. Blood and plasma taurine and serum cobalamin concentrations were measured. RESULTS: People who fed vegetarian diets to their cats did so largely for ethical considerations and were more likely than people who fed conventional diets to believe that there are health benefits associated with a vegetarian diet and that conventional commercial cat foods are unwholesome. Both groups were aware of the potential health problems that could arise from improperly formulated vegetarian diets. All cats evaluated had serum cobalamin concentrations within reference range, and 14 of 17 had blood taurine concentrations within reference range. CONCLUSIONS AND CLINICAL RELEVANCE: Vegetarian diets are fed to cats primarily for ethical considerations. Results of this study should aid practitioners in communicating with and providing advice to such clients.     

Relationships between Low Serum Cobalamin Concentrations and Methlymalonic Acidemia in Cats

Background: Serum cobalamin concentrations below reference range are a common consequence of gastrointestinal disease in cats. Serum cobalamin ≤ 100 ng/L is associated with methylmalonic acidemia.
Objectives: To determine the prevalence of cobalamin deficiency, defined by elevated serum methylmalonic acid (MMA), in cats with serum cobalamin ≤ 290 ng/L, and the optimum serum cobalamin concentration to predict cobalamin deficiency in cats.
Sample Set: Residual serum samples (n = 206) from cats with serum cobalamin ≤ 290 ng/L.
Methods: Retrospective, observational study. Serum cobalamin and folate were measured with automated assays. Serum MMA was determined by gas chromatography-mass spectrometry. Cobalamin deficiency was defined as serum MMA > 867 nmol/L. Sensitivity and specificity of serum cobalamin concentrations ≤290 ng/L for detecting MMA > 867 nmol/L were analyzed using a receiver-operator characteristic curve.
Results: There was a negative correlation between serum cobalamin and MMA concentrations (Spearman's r =−0.74, P < 0.0001). The prevalence of MMA ≥ 867 nmol/L in cats with serum cobalamin ≤ 290 ng/L was 68.4%. Serum cobalamin ≤ 160 ng/L had a 74% sensitivity and 80% specificity for detecting MMA > 867 nmol/L. No significant difference in serum folate concentrations was detected between affected and unaffected cats.
Conclusions and Clinical Importance: Elevated MMA concentrations, suggesting cobalamin deficiency, are common in cats with serum cobalamin ≤ 290 ng/L. Cobalamin deficiency is clinically significant, and supplementation with parenteral cobalamin is recommended for cats with gastrointestinal disease and low serum cobalamin concentrations.     

Cobalamin, folate and inorganic phosphate abnormalities in ill cats

Hypocobalaminaemia in cats has previously been identified, but the incidence reported has varied, and the frequency of folate deficiency is unknown. The aims of this study were to evaluate the incidence of low cobalamin and folate levels in a population of cats that were suffering predominantly from diseases of the alimentary tract (including the liver and pancreas) and to ascertain whether severity of disease (as assessed by bodyweight and body condition score (BCS)) related to degree of deficiency. The study population comprised 103 cats, of which 16.5% had low cobalamin levels and 38.8% had low folate levels. A serendipitous finding was inorganic phosphate levels below the reference range in 48% of the cases. Significant associations were found between subnormal cobalamin levels and median BCS (P=0.049); combined low folate and low cobalamin and bodyweight (P=0.002), BCS (P=0.024) and inorganic phosphate levels (P=0.003). The finding of low levels of folate and cobalamin in clinical cases suggests that supplementation may be indicated more frequently than is currently recognised.     

Hyperammonaemic encephalopathy secondary to selective cobalamin deficiency in a juvenile Border collie

An eight-month-old Border collie was presented with anorexia, cachexia, failure to thrive and stupor. Laboratory tests demonstrated a mild anaemia, neutropenia, proteinuria and hyperammonaemia. Serum bile acid concentrations were normal, but an ammonia tolerance test (ATT) was abnormal. The dog responded to symptomatic therapy for hepatoencephalopathy. When a low serum cobalamin (vitamin B12) concentration and methylmalonic aciduria were noted, the dog was given a supplement of parenteral cobalamin. Two weeks later, a repeat ATT was normal. Cobalamin supplementation was continued every two weeks, and all clinical signs, except for proteinuria, resolved despite withdrawing all therapy for hepatoencephalopathy. A presumptive diagnosis of hereditary selective cobalamin malabsorption was made, based on the young age, Border collie breed, low serum cobalamin concentration and methylmalonic aciduria. Although hereditary selective cobalamin malabsorption in Border collies, giant schnauzers, Australian shepherd dogs and beagles has previously been reported in North America, to the authors' knowledge this is the first report of the condition in the UK and the first to document an abnormal ATT in a cobalamin-deficient dog.     

Subnormal concentrations of serum cobalamin (vitamin B12) in cats with gastrointestinal disease

The present study sought to determine the spectrum of diseases associated with subnormal concentrations of serum cobalamin in cats undergoing investigation of suspected gastrointestinal problems. The solid-phase boil radioassay (RA) for cobalamin employed in the present study was immunologically specific, precise, and accurate, with a sensitivity of 15 pg/mL. The RA yielded results that strongly correlated with those obtained by bioassay (Spearmann rho = .805; P < .0001), although the absolute values were lower for the RA. Forty-nine of 80 serum samples submitted during the period of January 1996-January 1998 had cobalamin concentrations below the reference range for healthy cats (range 900-2,800 pg/mL; mean +/- SD, 1,775 +/- 535 pg/mL; n = 33). Cats with subnormal cobalamin concentrations (mean +/- SD; 384 +/- 272 pg/mL, range 3-883 pg/mL) were middle-aged or older and were presented for weight loss. diarrhea, vomiting, anorexia, and thickened intestines. Definitive diagnoses in 22 cats included inflammatory bowel disease (IBD), intestinal lymphoma, cholangiohepatitis or cholangits, and pancreatic inflammation. Serum concentrations of cobalamin were particularly low in cats with intestinal lymphoma, three-fifths of whom also had subnormal serum concentrations of folate (< 9 ng/mL). The simultaneous presence of disease in the intestines, pancreas, or hepatobiliary system in many cats made it difficult to determine the cause of subnormal cobalamin concentrations. The circulating half-life of parenteral cyanocobalamin was shorter in 2 cats with IBD (5 days) than in 4 healthy cats (12.75 days). The presence of subnormal serum concentrations of cobalamin in 49 of 80 cats evaluated suggests that the measurement of serum cobalamin may be a useful indirect indicator of enteric or pancreatic disease in cats. The rapid depletion of circulating cobalamin in cats suggests that cats may be highly susceptible to cobalamin deficiency. However, the relationship of subnormal serum cobalamin concentrations to cobalamin deficiency and the effect of cobalamin deficiency on cats remain to be determined.     

Association between serum cobalamin and methylmalonic acid concentrations in dogs

The aim of this study was to evaluate the association between serum methylmalonic acid (MMA), a proposed marker of cellular cobalamin deficiency, and serum cobalamin concentrations in dogs. Serum samples from 555 dogs were grouped according to their serum cobalamin concentrations (<150 ng/L to 1000 ng/L). Additionally, serum samples were collected from 43 healthy dogs to calculate a reference interval for canine serum MMA. MMA was measured using a GC/MS method. Groups were compared using a Kruskal-Wallis test with Dunn's post test. Proportions of dogs above the upper limit of the reference interval were calculated and a χ2-test for trend was performed to evaluate the association between serum cobalamin and MMA concentrations. The reference interval for serum MMA was calculated to be 414.7-1192.5 nmol/L. Dogs with serum cobalamin concentrations <251 ng/L had significantly higher MMA concentrations (P<0.05) and the χ2-test for trend showed a trend for increasing serum MMA concentrations with decreasing serum cobalamin concentrations (P<0.0001). Additionally, a number of dogs with normal serum cobalamin concentrations had increased serum MMA concentrations, suggesting that some of these dogs may have cobalamin deficiency on a cellular level. Further studies are warranted to determine if these dogs should receive cobalamin supplementation.     

Serum cobalamin concentrations in dogs with leishmaniosis before and during treatment

Hypocobalaminemia in dogs is most commonly associated with gastrointestinal disorders leading to impaired absorption and utilization of cobalamin. The objectives of this study were to compare serum cobalamin concentrations between dogs with leishmaniosis and clinically healthy dogs, and to assess possible alterations of serum cobalamin concentrations in dogs with leishmaniosis at different timepoints during treatment. Fifty-five dogs with leishmaniosis and 129 clinically healthy dogs were prospectively enrolled. Diagnosis of leishmaniosis was based on clinical presentation, positive serology and microscopic detection of Leishmania amastigotes in lymph node aspiration smears. Twenty of the dogs with leishmaniosis were treated with a combination of meglumine antimonate and allopurinol for 28 days and serum cobalamin concentrations were measured in blood samples that were collected before initiation of treatment (timepoint 0) and on days 14 and 28. In order to estimate alterations of serum cobalamin concentrations during treatment, cobalamin concentrations were measured in blood samples from 20 out of 55 dogs with leishmaniosis at all timepoints. Serum cobalamin concentrations were significantly lower in dogs with leishmaniosis before treatment (median: 362 ng/L; IQR: 277−477 ng/L) compared to clinically healthy dogs (median: 470 ng/L; IQR: 367−632 ng/L; P = 0.0035). Serum cobalamin concentrations increased significantly in dogs with leishmaniosis on day 14 of treatment compared to timepoint 0 (P = 0.02).In the present study, serum cobalamin concentrations were significantly lower in dogs with leishmaniosis compared to clinically healthy dogs. In addition, there was an increase in serum cobalamin concentrations during treatment. The clinical significance of hypocobalaminemia in dogs with leishmaniosis remains to be determined.     

The prevalence of hypocobalaminaemia in cats with spontaneous hyperthyroidism

Objectives : To determine the prevalence of hypocobalaminaemia in cats with moderate to severe hyperthyroidism and to investigate the relationship between cobalamin status and selected haematologic parameters. Methods : Serum cobalamin concentrations were measured in 76 spontaneously hyperthyroid cats [serum thyroxine (T₄) concentration ≥100 nmol/L] and 100 geriatric euthyroid cats. Erythrocyte and neutrophil counts in hyperthyroid cats with hypocobalaminaemia were compared with those in hyperthyroid cats with adequate serum cobalamin concentrations (≥290 ng/L). Results : The median cobalamin concentration in hyperthyroid cats was lower than the control group (409 versus 672 ng/L; P=0·0040). In addition, 40·8% of hyperthyroid cats had subnormal serum cobalamin concentrations compared with 25% of controls (P=0·0336). Weak negative correlation (coefficient: –0·3281) was demonstrated between serum cobalamin and T₄ concentrations in the hyperthyroid population, and the median cobalamin concentration was lower in cats with T₄ above the median of 153 nmol/L compared with cats with T₄ below this value (P=0·0281). Hypocobalaminaemia was not associated with neutropenia or anaemia in hyperthyroid cats. Clinical Significance : This study indicates that a substantial proportion of cats with T₄≥100 nmol/L are hypocobalaminaemic and suggests that hyperthyroidism directly or indirectly affects cobalamin uptake, excretion or utilisation in this species.     

Partial characterization of cobalamin deficiency in Chinese Shar Peis

A total of 22,462 serum sample results from dogs being evaluated for gastrointestinal disease at the Gastrointestinal Laboratory, College of Veterinary Medicine, Texas A&M University were evaluated retrospectively. The proportion of dogs with serum cobalamin concentrations below the reference interval and median serum concentrations were compared between Shar Peis and other dog breeds. Serum samples were also obtained prospectively from 22 healthy and 32 Shar Peis with chronic gastrointestinal disease and 59 healthy dogs of other breeds, and serum concentrations of cobalamin, folate, and methylmalonic acid were determined and compared. Overall, 64.0% (89/139) of serum samples from Shar Peis showed serum cobalamin concentrations below the limit of the reference interval and 38.1% (53/139) of these were below the detectable limit for the assay. The median serum cobalamin concentration in Shar Peis was significantly lower than in other breeds. Shar Peis with gastrointestinal disease had significantly lower serum cobalamin and higher serum methylmalonic acid concentrations compared to healthy Shar Peis. Healthy Shar Peis had significantly increased serum methylmalonic acid concentrations compared to healthy dogs of other breeds. There were no meaningful differences in folate concentrations between groups. In conclusion, Shar Peis have a high prevalence of cobalamin deficiency compared to other breeds and healthy Shar Peis may have subclinical cobalamin deficiency.     

Serum folate,cobalamin, homocysteine and methylmalonic acid concentrations in pigs with acute,chronic or subclinical Lawsonia intracellularis infection

Lawsonia intracellularis is the causative agent of porcine proliferative enteropathy. The clinical presentation can be acute (i.e. proliferative hemorrhagic enteropathy, PHE), chronic (i.e. porcine intestinal adenomatosis, PIA) or subclinical. In humans with chronic enteropathies, low serum folate (vitamin B₉) and cobalamin (vitamin B₁₂) concentrations have been associated with increased serum concentrations of homocysteine and methylmalonic acid (MMA), which reflect the availability of both vitamins at the cellular level. The aim of this study was to evaluate serum folate, cobalamin, homocysteine and MMA concentrations in serum samples from pigs with PHE, PIA or subclinical L. intracellularis infection, and in negative controls. Serum folate, cobalamin, homocysteine and MMA concentrations differed significantly among pigs in the PHE, PIA, subclinical and negative control groups. Serum folate concentrations in the PHE and PIA groups were lower than in the subclinical and negative control groups, while serum cobalamin concentrations were lower in the PIA group than in other groups. Serum concentrations of homocysteine were higher in the PHE, PIA and subclinical groups than in the negative control group. Serum concentrations of MMA were higher in the subclinical and PIA groups than in the control group. These data suggest that pigs infected with L. intracellularis have altered serum cobalamin, folate, homocysteine and MMA concentrations.     

Metabolism of amino acids in cats with severe cobalamin deficiency.

OBJECTIVE: To validate an automated chemiluminescent immunoassay for measuring serum cobalamin concentration in cats, to establish and validate gas chromatography-mass spectrometry techniques for use in quantification of methylmalonic acid, homocysteine, cysteine, cystathionine, and methionine in sera from cats, and to investigate serum concentrations of methylmalonic acid, methionine, homocysteine, cystathionine, and cysteine as indicators of biochemical abnormalities accompanying severe cobalamin (vitamin B12) deficiency in cats. SAMPLE POPULATION: Serum samples of 40 cats with severe cobalamin deficiency (serum cobalamin concentration < 100 ng/L) and 24 control cats with serum cobalamin concentration within the reference range. PROCEDURE: Serum concentrations of cobalamin were measured, using a commercial automated chemiluminescent immunoassay. Serum concentrations of methylmalonic acid, methionine, homocysteine, cystathionine, and cysteine were measured, using gas chromatography-mass spectrometry, selected ion monitoring, stable-isotope dilution assays. RESULTS: Cats with cobalamin deficiency had significant increases in mean serum concentrations bf methylmalonic acid (9,607 nmol/L), compared with healthy cats (448 nmol/L). Affected cats also had substantial disturbances in amino acid metabolism, compared with healthy cats, with significantly increased serum concentrations of methionine (133.8 vs 101.1 micromol/L) and significantly decreased serum concentrations of cystathionine (449.6 vs 573.2 nmol/L) and cysteine (142.3 vs 163.9 micromol/L). There was not a significant difference in serum concentrations of homocysteine between the 2 groups. CONCLUSIONS AND CLINICAL RELEVANCE: Cats with gastrointestinal tract disease may have abnormalities in amino acid metabolism consistent with cobalamin deficiency. Parenteral administration of cobalamin may be necessary to correct these biochemical abnormalities.     

Chronic-idiopathic enteropathy in cats--a case control study

Aim of this study is to clinically, microbiologically and histopathologically characterize inflammatory bowel disease (IBD) in the cat. Nine cats with chronic persistent or intermitent vomitus and diarrhea were examined between 1998 and 2001. All cats had a thuorough diagnostic workup performed. Full thickness biopsies from stomach, duodenum, jejunum, ileum and colon were surgically obtained for histopathological examination. Duodenal juice was obtained by direct aspiration for microbiologic qualitative and semiquantitative examination. Seven cats euthanized for other medical reasons were used as controls. Six cats had a lymphoplasmacytic IBD and three an eosinophilic IBD. Four cats with IBD had additional diseases diagnosed. Three cats with IBD had elevated bacterial counts. Retrospectively no correlation could be found between clinical symptoms and histopathological results. Serum TLI was not able to differentiate chronic pancreatitis and IBD. Serum folic acid and cobalamin did not correlate with the distribution of lesions in the gastrointestinal tract. Finally, no correlation was found between bacterial counts in the proximal duodenum and IBD.     

Effect of exocrine pancreatic insufficiency on cobalamin absorption in dogs

The possibility that the canine pancreas might have an important role in the physiologic absorption of cobalamin (vitamin B12) has been explored by determining the effect of exocrine pancreatic insufficiency on cobalamin absorption and by examining the subsequent influence of bovine pancreatic enzymes and canine pancreatic juice. Exocrine pancreatic insufficiency was induced by ligation of pancreatic ducts and confirmed by indirect assessment of exocrine pancreatic function. Cobalamin absorption was determined by oral administration of cyano[58Co]cobalamin and quantitation of radioactivity in blood, urine, and feces during 48 hours. Pancreatic duct ligation resulted in a significant (P less than 0.001) decrease in cobalamin absorption, which was not restored by oral administration of bovine pancreatic enzymes, despite considerable improvements in steatorrhea and in vivo proteolytic activities. In marked contrast, malabsorption of cobalamin was significantly (P less than 0.05) reversed by oral administration of canine pancreatic juice. These results indicate that pancreatic secretions have an important role in the normal absorption of cobalamin in the dog, a role that does not appear to be attributable to pancreatic enzymes, but is consistent with the existence of a pancreatic intrinsic factor in this species.     

Cobalamin deficiency associated with erythroblastic anemia and methylmalonic aciduria in a border collie.

Anemia due to cobalamin deficiency is a rare genetic disorder that has been recognized in dogs only recently. This report concerns a 14-month-old border collie that presented for chronic, nonregenerative anemia. Cytological examination of a peripheral blood smear showed the presence of erythroblasts. Serum cobalamin levels were below reference ranges reported for clinically normal dogs. A methylmalonic aciduria was found on urinalysis. These signs are consistent with the anemia in Imerslund-Graesbeck syndrome reported in humans. Anemia due to cobalamin deficiency responds to parenteral vitamin B12 therapy, and affected animals have a good prognosis for recovery.     

Validation of a radioassay for the determination of serum folate and cobalamin concentrations in dogs

Determinations of serum folate and cobalamin concentrations in dogs have proved of considerable value for the identification and characterisation of chronic small intestinal disorders, but the microbiological assays used are time-consuming and technically demanding. Dual isotope radio-assays are more convenient and have been developed for the determination of folate and cobalamin in human beings. This study has evaluated such an assay for the determination of serum folate and cobalamin concentrations in dogs by direct comparison with microbiological assays used previously. Assays were performed on samples from 77 dogs, including controls and animals with confirmed or suspected chronic small intestinal disease. Regression analysis demonstrated a significant relationship between the two assays for serum folate concentrations (R=0–85; P=0–0001) and a definite trend for radioassay to give lower results than bioassay. There was also a significant relationship between bioassay and radioassay data for serum cobalamin (R=0–91; P=0–0001) with comparable absolute values for these two assays. Radioassay of serum samples from 31 clinically healthy dogs gave control ranges of 3–7 to 8-8 4mUg/litre for folate and 205 to 490 ng/litre for cobalamin. These ranges were similar to those calculated by comparison with the established ranges for bioassay using regression analysis, which predicted ranges of 4-4 to 8-4 μg/litre and 217 to 398 ng/litre for radioassay of serum folate and cobalamin, respectively. These data indicate that a dual isotope radioassay of serum folate and cobalamin may be used for dogs, and emphasise the need for laboratories to validate and establish their own control ranges for different assays.