The effects of haemolysis on serum chemistry measurements in poultry

Abstract

The effects of haemolysis on serum chemistry values in broiler and layer chickens, and in turkeys were determined using two types of serum chemistry analysers, a wet reagent analyser and a dry slide reagent analyser. The interfering effects of haemolysis were evaluated using eight levels of haemoglobin in serum analysed by the wet reagent instrument and six levels of haemoglobin in serum analysed by the dry slide reagent instrument. Nine serum chemistry analytical tests were performed on each analyser, including determination of glucose, total protein, albumin, creatine kinase, gamma-glutamyl transferase, aspartate aminotransferase, calcium, phosphorus and uric acid. The interfering effects of haemolysis varied depending on type of analyser, type of bird and the specific test. With the wet reagent chemistry analyser, the gamma-glutamyl transferase, phosphorus and uric acid analytes were most sensitive to haemoglobin interference, and the albumin, total protein and creatine kinase analytes were most resistant. With the dry slide reagent analyser, the gamma-glutamyl transferase, phosphorus, and albumin analytes were most sensitive to haemoglobin interference, and the glucose and aspartate aminotransferase analytes were most resistant. The effects of haemoglobin interference were not consistent from one type of chemistry analyser to another. The dry slide reagent analyser did not appear to resist the effects of haemoglobin interference better than the wet reagent analyser in this study. Our results suggest the need to construct interferographs for each chemistry analyser, species, and type of bird.

Effect of Hemolysis on Plasma Protein Levels and Plasma Electrophoresis in Birds

“…Total protein concentration measurement may therefore be prone to both addition and optical interferences, because hemoglobin itself is a protein (Pontet, 2000), and the Biuret reaction used to carry on this measurement was based on an endpoint assay read at 553 nm. This phenomenon has been quite well documented in birds (Andreasen et al, 1996(Andreasen et al, , 1997. Hemolysis is described to induce artifactually higher total protein values (Lippi et al, 2006), even if this analysis is not extremely sensitive to interference by hemoglobin (Andreasen et al, 1996(Andreasen et al, , 1997.…”

“…As described in other studies on avian protein electrophoresis, the A/G ratio was calculated by dividing the sum of prealbumin and albumin by the sum of the globulin fractions (Lumeij, 1987;Cray and Tatum, 1998). Effect of hemolysis in electrophoresis patterns was investigated by comparing AUCs for each fraction rather than protein concentration values, because in hemolyzed samples total protein concentrations are often erroneous (Andreasen et al, 1996(Andreasen et al, , 1997.…”

“…Hemolysis is recognized as an important interference factor in human laboratory medicine (Kroll and Elin, 1994); the release of hemoglobin and other intracellular components from erythrocytes can alter electrophoretic results. Release of hemoglobin usually gives a characteristic pink-to-red tinge to the plasma or the serum (Kirschbaumweg, 2001;Lippi et al, 2006) and is a common phenomenon in both human and veterinary medicine (Andreasen et al, 1996(Andreasen et al, , 1997Benlakehal et al, 2000;Thomas, 2001). In birds, in vivo hemolysis is relatively rare (0.3% of the samples) and can occur as a result of acute lead toxicosis or the ingestion of crude oil by shore birds (Fry and Addiego, 1987;Fudge, 2000).…”

“…Published information of the effect of hemolysis on serum chemistry for birds is limited to two studies (Andreasen et al, 1996(Andreasen et al, , 1997. In humans and birds, hemolysis can lead to artifactual high values for plasma or serum total protein concentration measurements (Andreasen et al, 1996(Andreasen et al, , 1997Pontet, 2000;Vermer et al, 2007). There is less information for the influence of hemolysis in agarose gel protein electrophoresis.…”

Profile of Serum Metabolites and Proteins of Broiler Breeders in Rearing Age

“…However, it is an important tool, as some metabolic disorders are difficult to detect only by clinical signs (Andreasen et al, 1996). It can also assist the monitoring of poultry health, the diagnosis and treatment of diseases, and to assess their health status (Schmidt et al, 2007).…”