High resolution protein electrophoresis of 100 paired canine cerebrospinal fluid and serum

This study was performed to investigate the diagnostic relevance of cerebrospinal fluid (CSF) high resolution electrophoresis. The laboratory technique was applied to 100 paired samples of canine CSF and serum, with paired samples tested during the same analytical run, as recommended in human medicine. Ninety four of the dogs had a neurological disease and 6 healthy dogs served as a control group. A strong linear correlation between CSF total protein concentration and the albumin quota (AQ) was found in the control group and in the inflammatory (infectious or noninfectious), neoplastic, and miscellaneous groups: AQ = 0.015 CSF total protein--0.102, r = 0.990. This correlation suggests that an increased CSF total protein concentration can be an indicator of blood brain barrier dysfunction. The highest median AQ value was found in the aseptic suppurative meningitis group, but no statistical differences were found between this and the other groups. The AQ, calculated with this technique, did not provide any additional information. Moreover, although unexpected, the electrophoretic profiles were not characteristic of any particular disease. In conclusion, this study did not confirm high resolution electrophoresis of paired CSF and serum samples to be a valuable ancillary diagnostic tool for canine neurological diseases.     

Detection of biclonal gammopathy by capillary zone electrophoresis in a cat and a dog with plasma cell neoplasia

Gammopathies associated with plasma cell neoplasms in a 15-year-old female spayed domestic shorthaired cat and a 9-year-old female spayed Rottweiler dog were evaluated by serum protein electrophoresis. In the cat, the plasma cell neoplasm was found in the liver and spleen, and an evaluable sample of bone marrow was not obtained. Some of the plasma cells had the morphologic appearance of flame cells. The paraprotein was confirmed as IgG based on agar gel immunodiffusion precipitation and both immunocytochemical and immunohistochemical staining. The dog had multiple myeloma with production of IgG and IgA paraproteins. In both cases, serum proteins were evaluated by 2 methods of protein electrophoresis: cellulose acetate electrophoresis (CAE) and capillary zone electrophoresis (CZE). In the cat and the dog, CAE showed a single large oligoclonal-like peak, which occurred in the γ-region in the cat and the β-γ-region in the dog, whereas CZE showed a biclonal gammopathy with 2 very close narrow spikes in the γ- and β-γ-regions in the cat and dog, respectively. In selected cases, CZE may be more effective than routine CAE in distinguishing oligoclonal from monoclonal or biclonal paraproteinemia.     

Effects of haemolysis,lipaemia, bilirubinaemia and fibrinogen on protein electropherogram of canine samples analysed by capillary zone electrophoresis

The possible interference of haemolysis, lipaemia, bilirubinaemia and fibrinogen on capillary zone electrophoresis of canine samples were studied. Solutions of haemoglobin, lipid and bilirubin were prepared and mixed with serum aliquots to make up samples containing different concentrations of the putative interferent substance. In addition, samples of serum and plasma were assayed to assess the influence of fibrinogen. Haemolysis and lipids produced a change in electropherogram morphology giving an interference peak located in the beta-2 region when haemoglobin was increased, and in the alpha-2 region when lipids were increased. A rise in concentration of these interferents caused an increase in the beta and alpha-2 fractions respectively, and a decrease in the other fractions. Bilirubin did not alter morphology but gave an increase in the albumin and alpha-1 and a decrease in the alpha-2 and beta-2 fractions. No differences were found between serum and plasma samples, and fibrinogen did not produce any additional peak.     

Stability of hemostatic proteins in canine fresh frozen plasma units

Abstract: The stability of hemostatic proteins, including coagulation factors II, VII, VIII, IX, and X and von Willebrand factor (vWf), in canine fresh frozen plasma (FFP) units stored for up to 1 year was studied. Plasma units from 7 donor dogs were subjected to 4 treatments following collection, including storage at −30°C for 3 months, 6 months, and 1 year and storage at −20°C for 6 months. Coagulant factor activity and vWf concentrations were measured at these times. Significant differences between prestorage and poststorage values were noted for factors VIII, IX, and X, and vWf. Differences seemed to be least pronounced for plasma stored for 3 months; however, a significant interaction between prestorage and poststorage differences and the 4 treatment groups could not be demonstrated. On the basis of factor content in the present study, 15–20 mL/kg of FFP stored for up to 1 year was capable of providing approximately 10–15 U/kg of vWf and factors VIII, IX, X, and II, whereas 10 mL/kg FFP provided at least 10 U/kg of factor VII.     

Protein electrophoresis of psittacine plasma

BACKGROUND: Although protein electrophoresis (EPH) has been widely applied in human and veterinary medicine, it has only recently been implemented in the analysis of avian samples. OBJECTIVE: The purpose of this study was to examine the application of protein EPH to the analysis of psittacine plasma samples. Our goals were to describe protein fraction mobility, establish reference intervals for some common species, determine the coefficient of variation (CV) of the chosen method, and examine the effects of sample handling and sample condition. METHODS: Heparinized plasma samples from several common psittacine species (minimum sample size 50 each) were examined using the Beckman Paragon system and SPEP-II gels. Total protein was measured by refractometry. Reference intervals (95%) were calculated by the rank methods. RESULTS: Fraction migration patterns were found to vary among common psittacine species. Day-to-day CV for the EPH fractions ranged from 2.2% to 10.5%; within-run CV ranged from 4.8% to 10.8%; and total CV ranged from 3.2% to 14.8%. The highest CV was noted for the poorly defined alpha-globulin fraction. Prolonged refrigeration, repeated freeze-thawing, hemolysis, and lipemia altered the results. CONCLUSIONS: Protein fractions from psittacine species were variable in terms of migration pattern and protein concentration, which necessitates the use of species-specific reference intervals. Avian protein electrophoretic patterns and values should be interpreted based on knowledge of the CV associated with the technique as well as on the effects of sample handling and condition.     

Effect of storage conditions on prothrombin time, activated partial thromboplastin time and fibrinogen concentration on canine plasma samples

The present study was to assess the effect of storage conditions on prothrombin time (PT), activated partial thromboplastin time (aPTT) and fibrinogen concentration in blood samples of healthy dogs. Thirty-five dogs of various breeds were included in the study. Citrated blood samples were obtained and plasma was divided into four aliquots to assess selected clotting parameters by means of a coagulometer. The first aliquot was analysed within 1 h after collection, while the remaining 3 were stored at 8℃ for 4, 8 and 24 h, respectively. One-way repeated measures analysis of variance documented a significant decreasing effect on PT at 24 h compared to 8 h and on fibrinogen concentration after 8 and 24 h compared to sampling time and at 4 and 24 h compared to 8 h post sampling. In conclusion, the results of this study indicate that only fibrinogen appears prone to significant decrease. In fact, aPTT is not substantially affected by refrigeration for at least 24 h post sampling and PT showed a statistical difference that does not necessary indicate biological significance as the results obtained were within reference intervals for the dog.     

A comparison of an enzymatic and a gas-chromatographic method for measuring the acetate concentration in the blood plasma of cattle

The concentration of acetate in blood plasma was measured by both a gas-liquid chromatographic method and an enzymatic method using acetyl-CoA synthetase. When acetate was measured enzymatically without previous protein precipitation, the apparent concentration was lower than when the concentration measured by either a gas-chromatographic method or by the enzymatic method after protein precipitation with perchloric acid and neutralization.     

Reference Intervals of Serum Protein Concentrations from Clinically Healthy Female Ragusana Donkeys (Equus asinus) Determined by Cellulose Acetate Electrophoresis

The aim of this study was to evaluate total serum protein concentration measured using biuret reaction and the protein fractions determined using acetate cellulose electrophoresis in Ragusana donkeys (Equus asinus). Blood samples were collected from 68 clinically healthy female donkeys by jugular venipuncture. The serum levels of total proteins were determined using biuret method, and the separation of proteins was performed using acetate cellulose electrophoresis. Coefficients of variation were also calculated for within-assay precision, and were found to be less than 5% for α- and β₁-globulins and 8% or less for albumin, β₂-, and γ-globulins. A total of five protein fractions were separated and quantified: albumin, α-, β₁-, β₂-, and γ-globulins. Data obtained from young and adult subjects were compared using the Mann–Whitney U test. Reference intervals (2.5%-97.5% quantiles) were determined for total proteins (50.0-84.0 g/L), albumin (16.2-36.6 g/L), α-globulins (4.85-19.5 g/L), β₁-globulins (2.25-10.35 g/L), β₂-globulins (3.30-14.85 g/L), γ-globulins (10.0-30.5 g/L), and albumin/globulin ratio (0.41-1.13). In relation to age, statistically significant differences were found in total protein concentration and γ-globulins. The results obtained in the present study contributed to establish reference intervals of serum protein fractions obtained using acetate cellulose electrophoresis in female Ragusana donkeys to be used by practitioners for health control.