Agarose gel serum protein electrophoresis in cats with and without lymphoma and preliminary results of tandem mass fingerprinting analysis

Background: Serum electrophoretic profiles in cats are poorly characterized with respect to the proteins that comprise the globulin fractions, and interpretation of the electrophoretograms is routinely done in the absence of information about identity of the proteins found within each fraction. Objectives: The aims of this study were to compare protein fractions separated by serum protein electrophoresis (SPE) in healthy cats and in cats with lymphoma and to confirm some component proteins in the major fractions following SPE using tandem mass fingerprinting analysis (TMFA). Methods: Total protein concentration was measured and agarose gel SPE performed on serum from 14 healthy cats and 14 cats with lymphoma. The absolute protein concentration within each fraction was compared between the 2 groups. Bands corresponding to the SPE fractions were excised from the gels of 2 control cats and 1 cat with lymphoma and analyzed by liquid chromatography coupled to mass spectrometry. Results were compared with sequences in the National Center for Biotechnology Information protein database. Results: Median albumin concentrations were significantly decreased and median β‐globulin concentrations were significantly increased in cats with lymphoma. Narrow electrophoretic spikes were present in the β/γ‐globulin fraction in 3 cats with lymphoma. Following TMFA, multiple proteins were identified in each fraction, and their mobility agreed with results from previous studies generated using alternative techniques. Inter‐α (globulin) inhibitor 4 was identified in feline serum for the first time. Conclusions: Cats with lymphoma had lower albumin and higher β‐globulin concentrations than did healthy cats. Despite limitations of one‐dimensional agarose gel SPE, TMFA provided preliminary data to confirm the protein components of the various fractions.     

Serum concentration of some acute phase proteins in naturally occurring canine babesiosis: a preliminary study

BACKGROUND: Serum concentrations of acute phase proteins can provide valuable diagnostic information in the detection, prognosis, or monitoring of disease. Information available on the acute phase response in naturally occurring canine babesiosis is limited. OBJECTIVE: The purpose of this investigation was to retrospectively evaluate serum concentrations of haptoglobin, C-reactive protein, and ceruloplasmin in dogs naturally infected with Babesia canis. METHODS: Haptoglobin, C-reactive protein, and ceruloplasmin concentrations were measured in serum samples from dogs with uncomplicated (n = 6) and complicated (n = 1) babesiosis and compared with 6 healthy dogs. RESULTS: Serum C-reactive protein and ceruloplasmin concentrations were significantly higher in dogs with babesiosis; however, serum haptoglobin concentration was significantly lower compared with control dogs (P <.01). CONCLUSIONS: Results of this study suggest that acute phase protein concentrations could be beneficial in the diagnosis and determination of the severity of babesiosis in dogs.     

Assessment of IgE binding to native and hydrolyzed soy protein in serum obtained from dogs with experimentally induced soy protein hypersensitivity

OBJECTIVE: To assess binding of IgE to native, whole hydrolyzed, and separated hydrolyzed fractions of soy protein in serum obtained from dogs with experimentally induced soy protein hypersensitivity. ANIMALS: 8 na?ve Beagles (6 experimentally sensitized to native soy protein and 2 control dogs). PROCEDURES: 6 dogs were sensitized against soy protein by administration of allergens during a 90-day period. After the sensitization protocol was completed, serum concentrations of soy-specific IgE were measured and intradermal skin tests were performed in all 6 dogs to confirm that the dogs were sensitized against soy protein. Serum samples from each sensitized and control dog underwent western blot analysis to assess the molecular mass band pattern of the different allergenic soy fractions and evaluate reactivities to native and hydrolyzed soy protein. RESULTS: In sera from sensitized dogs, a characteristic band pattern with 2 major bands (approx 75 and 50 kd) and 2 minor bands (approx 31 and 20 kd) was detected, whereas only a diffuse band pattern associated with whole hydrolyzed soy protein was detected in the most reactive dog. Reactivity was evident only for the higher molecular mass peptide fraction. In control dogs, no IgE reaction to native or hydrolyzed soy protein was detected. CONCLUSIONS AND CLINICAL RELEVANCE: Data suggest that the binding of soy-specific IgE to the hydrolyzed soy protein used in the study was significantly reduced, compared with binding of soy-specific IgE to the native soy protein, in dogs with experimentally induced soy hypersensitivity.     

Analysis of mouse, rat, dog, marmoset, and human serum proteins by capillary electrophoresis: comparison with agarose gel electrophoresis

BACKGROUND: Serum protein analysis in both humans and experimental animal species has so far been carried out by labor-intensive techniques, such as agarose gel electrophoresis (AGE). OBJECTIVE: The objective of this study was to evaluate capillary electrophoresis (CE) as an alternative technique to AGE for the analysis of serum proteins from healthy animals. METHODS: Blood samples were collected into tubes without anticoagulant from 6 fasted healthy male mice, rats, dogs, marmosets, and humans. Serum proteins were separated by CE using a technique standardized for the analysis of human proteins, and the results (efficiency, resolution, and precision) were compared with those obtained through AGE. RESULTS: Compared with AGE, CE resulted in narrower peaks and more peaks. The efficiency of protein separation by CE was significantly higher for all species, and resolution (R) was significantly higher in samples from dogs. Using rat serum, intraday reproducibility was lower for all protein fractions, and interday reproducibility was lower for most peaks, compared with AGE. CONCLUSIONS: We conclude that CE is a viable alternative to AGE for the determination of protein electrophoresis in a routine veterinary clinical pathology laboratory. The minimal sample requirement (2 microL), complete automation, and quantitative results make CE an especially valuable technique for protein analysis in experimental animal models.     

Serum proteins in normal cats and cats infected with Aelurostrongylus abstrusus

Serum proteins were evaluated by cellulose acetate electrophoresis in cats prior to and every 2 weeks for 24 weeks after oral infection with third stage larvae of Aelurostrongylus abstrusus. Evaluation of electrophoretograms was standardized by determination of electrophoretic migration ratios. Six fractions of serum proteins were consistently identified: albumin and alpha 1, alpha 2, beta 1, beta 2, and gamma globulin. Relative and absolute concentrations of each serum protein fraction were determined. The only changes found were a decrease in concentration of alpha globulins and an increase in concentration of beta 1 globulins. These changes were mild, however, so that the the concentrations for infected cats were still within 1 SD of the control concentrations. Apparently serum electrophoresis is not a useful diagnostic test for aelurostrongylosis.     

Serum protein electrophoresis in retired racing Greyhounds

BACKGROUND: Retired racing Greyhounds are becoming common as pets. Because of their unique physiology, results of routine laboratory tests are frequently outside the reference interval for dogs. Compared with other breeds, Greyhounds have low serum protein concentrations, but the concentrations of different serum protein fractions have not been reported. OBJECTIVES: Our objectives were to evaluate the results of serum protein electrophoresis (SPE) in healthy, retired racing Greyhounds and compare them with a control group of age- and gender-matched non-Greyhound dogs. METHODS: Agarose gel electrophoresis was done using a standard method; the gels were stained with amido black and scanned with a Cliniscan 2 densitometer (Helena Laboratories, Beaumont, TX, USA). Protein fractions were identified by visual inspection of the electrophoretogram. A Student's t-test assuming equal variances was used to compare the concentration of the different fractions between groups. RESULTS: The concentrations of total protein, total globulins, and alpha-1-, alpha-2-, beta-1-, and beta-2-globulins were significantly lower and the albumin to globulin (A:G) ratio was significantly higher in Greyhounds than in non-Greyhound dogs (P < .05). There was no significant difference in albumin or gamma-globulin concentrations. CONCLUSIONS: Low serum protein concentrations in Greyhounds are the result of low concentrations of a- and b-globulins. These results should be kept in mind when evaluating both healthy and sick Greyhounds. Additional studies are needed to identify the individual proteins associated with low alpha- and beta-globulin concentrations in Greyhounds.     

Separation of serum glycated proteins by agarose gel electrophoresis and nitroblue tetrazolium staining in diabetic and normal cats

BACKGROUND: The total glycated protein (fructosamine) concentration in serum consists mainly of glycated albumin and lipoproteins. Measurement of fructosamine is used to diagnose and monitor diabetes mellitus in cats. OBJECTIVE: The aims of this study were to measure glycated proteins in diabetic and healthy (nondiabetic) cats using a semiquantitative technique and to determine whether measurement of any of the fractions of glycated protein could be potentially advantageous for the diagnosis and monitoring of diabetic cats. METHODS: Serum samples from 6 cats with diabetes mellitus and 10 clinically healthy adult cats were assayed for total glycated protein using a nitroblue tetrazolium (NBT) fructosamine assay. Serum proteins were separated by agarose gel electrophoresis and stained with NBT to identify individual glycated proteins within the bands. Gels were scanned by densitometry at 525 nm and the glycated protein content was calculated with reference to the total glycated protein content of the sample. RESULTS: Diabetic cats with increased total fructosamine concentrations had higher concentrations of glycated albumin and glycated alpha- and beta-lipoproteins compared with healthy cats. The concentration of glycated proteins in each of the fractions had a positive linear association with the total glycated protein content of serum, but there was large variation in the relative contributions of the 3 protein fractions to the total glycated protein concentration. CONCLUSIONS: Based on the results of this study, measurement of individual glycated fractions does not seem to offer any potential diagnostic advantage over measurement of total glycated protein (fructosamine) concentration alone. In some diabetic and healthy cats, glycated lipoproteins formed the major part of the total glycated protein, whereas in other cats albumin was the major contributor.     

Assessment of serum protein electrophoresis for monitoring therapy of naturally acquired equine cyathostomin infections

Serum protein electrophoresis (SPE) has previously been suggested as a means of assessing cyathostomin burdens in horses, although SPE used for that purpose is supported by little evidence. This clinical research report describes a study that objectively evaluated the use of SPE on a population of 38 horses following the administration of different anthelmintics. The population was subdivided into three groups, Groups F, M and P: 7.5 mg/kg bwt fenbendazole was administered to Group F on day -12; on day 0 0.4 mg/kg bwt moxidectin was administered to Group M and 19 mg/kg bwt pyrantel was administered to Group P. Faecal worm egg counts were obtained on days -14, 0 and 10. Groups M and P acted as controls for the Group F faecal egg count reduction test (FECRT) in which a high level of benzimidazole resistance was demonstrated. Group F was then used as a control group for the FECRT for both Groups P and M. A high anthelmintic efficacy of moxidectin and pyrantel was detected. SPE was performed on venous blood collected on days 0, 10, 30, 56 and 80. As the cyathostomins infecting the horses had been shown to be highly resistant to fenbendazole, Group F then served as a control group for comparison of any changes in protein fractions. Serum proteins did not vary significantly between groups on any of the sampling dates. No significant changes in serum proteins were observed in any group and no patterns were apparent on qualitative assessment of SPE profiles. SPE was therefore concluded to be an insensitive tool for the monitoring of cyathostomin treatment in horses in a clinical environment.     

Evaluation of variation in serum globulin concentrations in dairy cattle

BACKGROUND: Several factors may influence the concentration of serum globulins in healthy cows and thereby affect clinical interpretation; however, few studies have addressed sources of variation in globulin values. OBJECTIVES: The purpose of this study was to compare colorimetry-based total serum globulin values with electrophoretically-determined serum globulin fractions and with IgG concentration, and to evaluate diurnal and long-term physiological variation and the effects of lactation and venipuncture site on serum globulin concentrations in Holstein dairy cattle. METHODS: Serum total globulin and albumin concentrations were analyzed by colorimetry and electrophoresis in 86 lactating cows; IgG concentrations were determined by radial immunodiffusion in 41 dry and 34 lactating cows. Serum globulins were analyzed hourly for 24 hours in 8 lactating cows and weekly for 15 weeks in 6 additional cows. Globulin concentrations were compared in samples obtained from jugular and coccygeal venipuncture sites in 4 cows. Results were analyzed using parametric statistical tests. RESULTS: Colorimetry-based total serum globulin concentrations correlated well with gamma-globulin fractions (r2 = 0.87) and IgG concentrations (r2 = 0.91). Diurnal variation of total serum globulins concentration was significant (P =.01); however, globulins did not vary significantly over a 15-week period. Mean serum globulins concentration in samples obtained from the jugular vein was 2.35 g/L higher than that in samples obtained by coccygeal venipuncture (P <.0001). CONCLUSIONS: The colorimetric method used widely in routine laboratory analyses remains a useful test for globulins determination in dairy cattle. However, time of sampling and venipuncture site should be considered in the interpretation of serum globulins on serial or interindividual specimens.     

Measurement of neonatal equine immunoglobulins for assessment of colostral immunoglobulin transfer: comparison of single radial immunodiffusion with the zinc sulfate turbidity test, serum electrophoresis, refractometry for total serum protein, and the sodium sulfite precipitation test.

Four procedures for assessment of adequacy of colostral immunoglobulin (Ig) transfer in foals were evaluated. Results of zinc sulfate turbidity test, serum electrophoresis, total serum protein refractometry, and sodium sulfite precipitation test were compared with immunoglobulin G content determined by single radial immunodiffusion. The zinc sulfate turbidity test gave acceptable results for IgG, except that hemolyzed serum samples gave higher than expected values. A correction factor for hemolyzed serum was found to be useful. Serum electrophoresis was a satisfactory method of estimating IgG content. Total serum protein values may not be a valid basis for estimating IgG content, inasmuch as postsuckling total protein values were found to decrease in some foals in which passive transfer of IgG had been adequate. Sodium sulfite precipitation reactions were too unpredictable to be of value for determination of neonatal IgG concentration.     

Serum Cardiac Troponin I Concentrations in Cattle with Cardiac and Noncardiac Disorders

Background: Making a clinical diagnosis of pericarditis in cattle is difficult and additional diagnostic tests are needed to evaluate cattle with suspected pericarditis. Serum cardiac troponin I (cTnI) concentrations are increased in cattle with pericarditis, but the utility of measuring serum cTnI concentrations in cattle with suspected pericarditis in cattle remains unclear.
Objectives: To determine if serum cTnI concentrations in cattle can be used to differentiate pericarditis from other cardiac disorders and noncardiac thoracic diseases.
Animals: Seventy-seven clinically diseased cattle and 19 healthy control cattle.
Methods: Serum cTnI concentrations were measured using an Immunlite Troponin I immunometric chemiluminescent assay in consecutive cases of postmortem-confirmed pericarditis (n = 18), endocarditis (n = 15), chronic suppurative pneumonia (n = 13), congenital heart disease (n = 10), reticulitis (n = 3), mediastinal abscess (n = 7), thymic lymphoma (n = 6), and caudal vena cava thrombosis (n = 5). Serum cTnI concentrations were measured in 19 healthy cattle.
Results: Although serum cTnI concentrations were significantly higher in cattle with pericarditis compared with healthy cattle, they were not significantly different from concentrations in cattle with endocarditis, congenital cardiac disease, mediastinal abscess, reticulitis, caudal vena cava thrombosis, or chronic suppurative pneumonia.
Conclusions: Serum cTnI cannot be used to distinguish cattle with pericarditis from cattle with other primary cardiac diseases. In addition, serum cTnI concentrations cannot distinguish between cattle with primary cardiac diseases and those with other noncardiac, intrathoracic disorders.     

Concentrations of serum amyloid A and lipopolysaccharide-binding protein in horses with colic

OBJECTIVE: To determine concentrations of 2 acute-phase proteins (serum amyloid A [SAA] and lipopolysaccharide-binding protein [LBP]) in serum samples obtained from horses with colic and identify relationships among these acute-phase proteins and clinical data. ANIMALS: 765 horses with naturally developing gastrointestinal tract diseases characterized by colic (ie, clinical signs indicative of abdominal pain) and 79 healthy control horses; all horses were examined at 2 university teaching hospitals. PROCEDURE: Serum concentrations of SAA and LBP were determined by immunoturbidometric and dot-blot assays, respectively. RESULTS: SAA and LBP concentrations were determined for 718 and 765 horses with colic, respectively. Concentrations of SAA were significantly higher in nonsurvivors than in survivors, and horses with enteritis or colitis and conditions characterized by chronic inflammation (eg, abdominal abscesses, peritonitis, or rectal tears) had SAA concentrations significantly greater than those for horses with other conditions. Serum concentrations of LBP did not correlate with outcome, disease process, or portion of the gastrointestinal tract affected. CONCLUSIONS AND CLINICAL RELEVANCE: Circulating concentrations of SAA were significantly higher at admission in horses with colic attributable to conditions having a primary inflammatory cause (eg, enteritis, colitis, peritonitis, or abdominal abscesses) and were higher in horses that failed to survive the episode of colic, compared with concentrations in horses that survived. Serum concentrations of LBP did not correlate with survival. Analysis of these findings suggests that evaluation of SAA concentrations may be of use in identifying horses with colic attributable to diseases that have inflammation as a primary component of pathogenesis.     

Unusual IgM-secreting multiple myeloma in a dog

A 4-year-old castrated male dog was evaluated because of multiple-limb lameness. Signs of pain were elicited during palpation of the regions of the proximal tibial metaphyses and distal left radial diaphysis. Radiography revealed osteolytic lesions of the long bones. Blood analyses revealed hypercalcemia and transient cytopenias. Serum protein electrophoresis did not reveal a monoclonal gammopathy; however, urine protein electrophoresis revealed Bence Jones proteinuria. Serial diagnostic sampling of bone lesions, immunohistochemical staining methods, and serum and urine protein immunoelectrophoresis were required to establish a diagnosis of multiple myeloma. Two IgM components were identified via serum protein immunoelectrofixation. The dog improved clinically after initiation of chemotherapy with melphalan and prednisone; however, the dog ultimately was euthanatized because of pathologic fracture. The case was unique because there was lack of vertebral involvement, an unusual gammopathy, and difficulty in identifying myeloma cells via serial sampling.     

Immunologic responses against hydrolyzed soy protein in dogs with experimentally induced soy hypersensitivity

OBJECTIVE: To assess whether dogs with experimentally induced type I hypersensitivity against soy protein would respond to soy hydrolysate and develop cutaneous or gastrointestinal tract reactions after intradermal and oral challenge exposure. ANIMALS: 12 na?ve Beagle pups (9 sensitized and 3 control dogs). PROCEDURE: 9 dogs were sensitized against soy protein by administration of allergens during a 90-day period. After the sensitization period, serum concentrations of soy-specific IgE were determined and an intradermal test was performed to confirm the dogs were sensitized against soy protein. An intradermal challenge test and an oral challenge test with native and hydrolyzed soy protein were conducted on 6 sensitized and 2 control dogs. RESULTS: High serum concentrations of soy-specific IgE and positive results for the intradermal test were observed for the 9 sensitized dogs after completion of the sesitization process. Sensitized dogs challenge exposed with hydrolyzed soy protein had a reduced inflammatory response after intradermal injection and no clinical response after an oral challenge exposure, compared with responses after intradermal and oral challenge exposure with native soy protein. CONCLUSIONS AND CLINICAL RELEVANCE: Soy-sensitized dogs did not respond to oral administration of hydrolyzed soy protein. Thus, hydrolyzed soy protein may be useful in diets formulated for the management of dogs with adverse reactions to food.     

Assessment of lipid peroxidation and serum vitamin E concentration in dogs with immune-mediated hemolytic anemia

OBJECTIVE: To determine plasma malondialdehyde (MDA) and serum vitamin E concentrations in dogs with immune-mediated hemolytic anemia (IMHA) and healthy control dogs. SAMPLE POPULATION: Serum and plasma samples from 36 dogs with IMHA and 40 healthy control dogs. PROCEDURE: Blood samples were collected from all study dogs. Plasma MDA concentrations were measured by use of a commercial colorimetric assay, and serum vitamin E concentrations (alpha-, gamma, and delta-tocopherol concentrations) were measured via high-performance liquid chromatography. RESULTS: Plasma MDA concentrations were significantly higher in the dogs with IMHA than in the control dogs. Compared with control dogs, serum alpha-, gamma-, and &tocopherol concentrations were significantly lower in the IMHA-affected dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated a state of oxidative stress and reduced antioxidant reserve in dogs with IMHA; this finding provides support for further investigation of the potential benefits of antioxidant treatment in dogs with this disease.     

Serum concentration of surfactant protein D in horses with lower airway inflammation

Reasons for performing study: Surfactant protein D (SP‐D), mainly synthesised by alveolar type II cells and nonciliated bronchiolar cells, is one important component of innate pulmonary immunity. In man, circulating concentrations of SP‐D are routinely used as biomarkers for pulmonary injury. To date, serum SP‐D levels have only been investigated in horses in an experimental model of bacterial airway infection. Objectives: To compare serum SP‐D concentrations at rest and after exercise in horses with and without inflammatory airway disease (IAD). Methods: Venous blood samples were collected from 42 Standardbred racehorses at rest and 60 min after performing a standardised treadmill exercise test. Tracheal wash and bronchoalveolar lavage fluid (BALF) samples were collected after exercise. Based on BALF cytology, 22 horses were defined as IAD‐affected and 20 classified as controls. Serum SP‐D concentrations were assessed using a commercially available ELISA kit and statistically compared between groups of horses and sampling times. Results: Serum concentrations of SP‐D in IAD‐affected horses were significantly higher than those of control horses, both at rest and after exercise. Within the IAD‐affected group, no significant correlation was found between serum SP‐D concentrations and BALF cytology. Within each group of horses (IAD and control), no significant influence of exercise was found on serum SP‐D levels. Conclusions: This is the first study determining serum SP‐D concentrations in a noninfectious, naturally occurring form of lower airway inflammation in horses. The results highlight that IAD is associated with a detectable, though moderate, increase of circulating SP‐D levels. Potential relevance: Serum concentration of surfactant protein D could represent a potentially valuable and readily accessible blood biomarker of equine lower airway inflammation.     

Prevalence and prognostic importance of hypomagnesemia and hypocalcemia in horses that have colic surgery

OBJECTIVE: To determine the prevalence of hypomagnesemia and hypocalcemia in horses with surgical colic. ANIMALS: 35 horses with surgically managed colic. PROCEDURE: Serum concentrations of total magnesium (tMg2+) and calcium (tCa2+), as well as ionized magnesium (iMg2+) and calcium (iCa2+) were analyzed before surgery and 1, 3, 5, and 7 days following surgery. A lead-II ECG and pertinent clinical data were also obtained at each time. RESULTS: Preoperative serum tMg2+ and iMg2+ concentrations were below the reference range in 6 (17%) and 19 (54%) horses, respectively. Serum concentrations of tCa2+ and iCa2+ were less than the reference range in 20 (57%) and 30 (86%) horses before surgery. Horses with strangulating lesions of the gastrointestinal tract had significantly lower preoperative serum concentrations of iMg2+ and iCa2+, as well as a higher heart rate than horses with nonstrangulating lesions. Horses that developed postoperative ileus had significantly lower serum concentrations of iMg2+ after surgery. Serum concentrations of magnesium and calcium (total and ionized) correlated significantly with the PR, QRS, QT, and corrected QT (QTc) intervals. Horses that were euthanatized at the time of surgery (n = 7) had significantly lower preoperative serum concentrations of iMg2+, compared with horses that survived. Neither serum magnesium nor calcium concentrations were predictors of hospitalization time or survival. CONCLUSIONS AND CLINICAL RELEVANCE: Hypomagnesemia and hypocalcemia were common during the perioperative period, particularly in horses with strangulating intestinal lesions and ileus. Serum concentrations of tMg2+ and tCa2+ were less sensitive than iMg2+ and iCa2+ in detecting horses with hypomagnesemia and hypocalcemia.     

Serum T3 and T4 concentrations in lambs with nutritional myodegeneration

BACKGROUND: Selenium (Se) deficiency is an important etiological factor in Nutritional Myodegeneration Disease (NMD) of lambs, and Se is required for synthesis of thyroid hormones. HYPOTHESIS: That serum T4 and T3 concentrations in lambs with NMD will be abnormal. ANIMALS: Ten healthy lambs and 15 lambs with NMD were included in the study. METHODS: Serum thyroid hormone concentrations were measured by chemiluminescence assay in samples collected from control and NMD lambs before and after treatment with a preparation containing sodium selenite, vitamin E, and vitamin B1, which was administered subcutaneously to lambs with NMD twice, with a 2-week interval. RESULTS: Before treatment, serum concentrations of fT3 and TT3 in lambs with NMD were lower than those of control group (P < .001), while serum concentrations of fT4 and TT4 in lambs with NMD were significantly higher than those of control group (P < .001, P < .01, respectively). Similarly, serum creatine kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) values in lambs with NMD were significantly higher compared with those of control (P < .001, P < .001, P < .001, P < .01, and P < .05, respectively) before treatment. However, serum concentrations of Se and alpha-tocopherol in lambs with NMD were significantly lower compared with those of control (P < .001). After treatment (at day 30), none of the variables were significantly different between control and NMD lambs (P > .05). CONCLUSIONS AND CLINICAL IMPORTANCE: Nutritional myodegeneration in lambs is associated with abnormalities in serum thyroid concentrations. Abnormalities in serum thyroid concentrations can result from selenium deficiency in NMD lambs. Serum thyroid concentrations together with serum CK, LDH, AST, ALT, and ALP values can be considered additional tools in diagnosis and prognosis of NMD in lambs.     

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.     

Serum protein concentrations from clinically healthy horses determined by agarose gel electrophoresis

Background: Serum protein electrophoresis is a useful screening test in equine laboratory medicine. The method can provide valuable information about changes in the concentrations of albumin and α‐, β‐, and γ‐globulins and thereby help characterize dysproteinemias in equine patients. Reference values for horses using agarose gel as a support medium have not been reported. Objectives: The purpose of this study was to establish reference intervals for serum protein concentrations in adult horses using agarose gel electrophoresis and to assess differences between warm‐blooded and heavy draught horses. In addition, the precision of electrophoresis for determining fraction percentages and the detection limit were determined. Methods: Blood samples were obtained from 126 clinically healthy horses, including 105 Thoroughbreds and 21 heavy draught horses of both sexes and ranging from 2 to 20 years of age. The total protein concentration was determined by an automated biuret method. Serum protein electrophoresis was performed using a semi‐automated agarose gel electrophoresis system. Coefficients of variation (CVs) were calculated for within‐run and within‐assay precision. Data from warm‐blooded and draught horses were compared using the Mann–Whitney U test. Results: Within‐run and within‐assay CVs were <5% for all protein fractions. No significant difference was found between warm‐blooded and heavy draught horses and so combined reference intervals (2.5–97.5%) were calculated for total protein (51.0–72.0 g/L), albumin (29.6–38.5 g/L), α₁‐globulin (1.9–3.1 g/L), α₂‐globulin (5.3–8.7 g/L), β₁‐globulin (2.8–7.3g/L), β₂‐globulin (2.2–6.0 g/L), and γ‐globulin (5.8–12.7 g/L) concentrations, and albumin/globulin ratio (0.93–1.65). Conclusion: Using agarose gel as the supporting matrix for serum protein electrophoresis in horses resulted in excellent resolution and accurate results that facilitated standardization into 6 protein fractions.