Plasma agarose gel electrophoresis (AGE) is recognized as a very reliable diagnostic tool in avian medicine. Within the last 10 years, new electrophoresis techniques such as capillary zone electrophoresis (CZE) have emerged in human laboratory medicine but have never been investigated in birds. To investigate the use of CZE in birds and to compare it with AGE, plasma samples from 30 roosters (Gallus gallus), 20 black kites (Milvus migrans), and 10 racing pigeons (Columba livia) were analyzed by both AGE and CZE. For the 3 species studied, values determined by AGE and CZE were well correlated for albumin and beta and gamma fractions whereas other values differed significantly. Values for alpha-3 fraction in the rooster, alpha-1 fraction in the black kite, and alpha fractions in the pigeon obtained by AGE were very well correlated with the prealbumin fraction values obtained by CZE. Repeatability and reproducibility appeared higher with CZE than with AGE. Although the interpretation of CZE electrophoresis patterns seems to produce results similar to those obtained with AGE, some proteins present in the alpha fraction measured with AGE migrated to the prealbumin fraction found with CZE. Although CZE requires the use of specific reference intervals and a much higher sample volume, this method has many advantages when compared with AGE, including better repeatability and reproducibility and higher analysis output.
Protein electrophoresis is recognized as a reliable diagnostic tool for birds even though results are seldom pathognomonic. Unfortunately, this technique is underused in avian medicine because many factors interfere with electrophoresis patterns; hemolysis is one of these factors and is often associated with improper specimen handling. In human laboratory medicine, hemolysis is a known interference factor that can lead to erroneous results. Published data on the influence of hemolysis on protein electrophoresis in birds is currently restricted to a single study in Psittacidae. The aim of this study was to further investigate this effect and to analyze potential interspecific differences. Blood samples were drawn from 28 Black Kites (Milvus migrans) and 19 Bar-headed Geese (Anser indicus) and separated into two aliquots. One aliquot was dipped into liquid nitrogen for 5 sec in order to cause freeze-thawing hemolysis before centrifugation. Total plasma protein concentration, plasma hemoglobin concentration, and plasma protein electrophoresis patterns were determined for both hemolyzed and nonhemolyzed samples. In both species, hemolysis resulted in falsely elevated total plasma protein concentration. In Bar-headed Geese, hemolysis caused a rise in the gamma fraction. In Black Kites, this rise involved not only the gamma fraction but also the beta fraction, stressing the potential for species-related differences. In both species, the effects of hemolysis mimicked a chronic inflammatory condition with resulting antigenic stimulation.
ABSTRACT:Plasma protein electrophoresis is recognized as a reliable diagnostic tool in avian medicine; however, the influence of circannual phenomena such as molt on protein electrophoregrams is poorly documented. The molt is a period of heavy hormonal and metabolic change in birds. The purpose of this study was to investigate the effects of molt on total protein concentration and electrophoresis patterns in birds. Blood samples were taken from 19 Barheaded Geese (Anser indicus) from mid-May to mid-August, at 15-day intervals. At the same time, molting stage of each bird was recorded. Total protein concentrations were measured and plasma agarose gel electrophoresis was performed on these samples. The Bar-headed Goose was chosen as a model, because they molt over a very short period. The total protein concentration and albumin, alpha-2, beta, and gamma fractions were at their minimum values during molt, whereas the prealbumin and alpha-1 fractions rose to their maximum levels. This study provides baseline information relevant to changes occurring in avian proteinograms throughout the molt. The increase in prealbumin and alpha-1 fractions may be related to an increase in plasma thyroid hormones during molt. The decrease observed in albumin, alpha-2, beta, and gamma fractions may be related to protein and energy shifts toward feather growth, as well as to an expansion of the circulatory system located around the feather follicles with secondary dilutional effects on protein fractions. From a clinical point of view, the observed changes associated with molting were less significant than initially expected, and would not likely results in incorrect diagnoses based on interpretation of the protein electrophoretic patterns.
Apo A-I accounts for the prominent alpha-globulin peak and comprises a major proportion of total protein concentration in diverse avian species. As a high-density lipoprotein and negative acute phase protein with a pivotal role in cholesterol homeostasis, further study is warranted to determine the significance of changes in apo A-I concentration in avian electrophoretograms.
The objective of this study was to evaluate the effects of stress (as measured by total heterophil + eosinophil counts [THECs] and plasma corticosterone [PC] levels) on plasma protein electrophoresis (PPE) in two Anseriform species. Ten red-breasted geese ( Branta ruficollis ) and eight Hawaiian geese (Branta sandvicensis) were received into quarantine at Beauval Zoo and housed together with other Anseriformes. Two days later, all were examined as part of routine quarantine procedures, and blood was collected. THECs were performed using a Malassez hemocytometer after a 1 : 200 dilution with an eosinophil dilution liquid containing phloxine B. PPE, as well as total protein and PC assays, was performed on lithium heparin plasma. Twenty days after arrival, all birds were similarly restrained, examined, and sampled. Pododermatitis lesions were identified on four geese that were excluded from statistical analysis. For each goose, THECs and PC values were sorted by value (lower or higher) independently from the day of sampling. A Wilcoxon signed rank test showed no significant differences between lower and higher values of THECs for any of the PPE fractions. Higher values of corticosterone were associated with higher values of prealbumin, but none of the other fractions were significantly different. A Spearman rank correlation coefficient showed that THECs and PC were not correlated; this suggested differences in kinetics between these stress markers. Results did not show significant alterations of electrophoresis patterns associated with stress as measured with THECs and PC. The results also highlight the complexity of precisely assessing acute and chronic stress in avian species.
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