Measurement of Redox Biomarkers in the Whole Blood and Red Blood Cell Lysates of Dogs

Abstract

The evaluation of the biomarkers of oxidative status is usually performed in serum, however, other samples, such as red blood cells (RBCs) lysates or whole blood (WB), can be used. The objective of this study was to evaluate if a comprehensive panel of redox biomarkers can be measured in the WB and RBCs of dogs, and their possible changes “in vitro” after the addition of different concentrations of ascorbic acid. The panel was integrated by biomarkers of the antioxidant status, such as cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of plasma (FRAP), Trolox equivalent antioxidant capacity (TEAC), thiol and paraoxonase type 1 (PON-1), and of the oxidant status, such as total oxidant status (TOS), peroxide-activity (POX-Act), reactive oxygen-derived compounds (d-ROMs), advanced oxidation protein products (AOPP) and thiobarbituric acid reactive substances (TBARS). All the assays were precise and accurate in WB and RBCs lysates. In addition, they showed changes after ascorbic acid addition that are in line with previously published results, being WB more sensitive to detect these changes in our experimental conditions. In conclusion, the panel of assays used in this study can be measured in the WB and RBCs of the dog. In particular, the higher sensitivity to detect changes in our experimental conditions and its easier sample preparation makes WB a promising sample for the evaluation of redox status in dogs, with also potential applications to other animal species and humans.

Changes in biomarkers of redox status in serum and saliva of dogs with hypothyroidism

“…This report should be considered a preliminary study, and, as previously mentioned, further trials with larger populations should be made to evaluate the changes in the oxidative status analytes used in this study throughout treatment. Also, an evaluation of the biomarkers in other sample types, such as whole blood and red blood cell lysates should be performed, since in a previous study these samples provided additional interesting information about the redox status in dogs [54]. In addition, further studies should be made to evaluate the applicability of these biomarkers in the diagnosis of this disease and treatment monitorization.…”

Changes in biomarkers of redox status in serum and saliva in dogs with hypothyroidism

“…This report should be considered a preliminary study, and, as previously mentioned, further trials with larger populations should be made to evaluate the changes of the oxidative status analytes used in this study throughout treatment. Also, an evaluation of the biomarkers in other sample types, such as whole blood and red blood cell lysates should be performed since in a previous study these samples provided additional interesting information about the redox status in dogs [54].…”

Effect of the Probiotic Bacillus subtilis DE-CA9TM on Fecal Scores, Serum Oxidative Stress Markers and Fecal and Serum Metabolome in Healthy Dogs

“…The determination of total thiol for this study was based on the reaction of thiols present in the sample with 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB). This assay was performed according to previously described protocols [ 36 ]. Results are expressed in micromoles per liter (µmol/L).…”

“…The measurement of total oxidant status (TOS) for this study is based on a assay described in a previously published methodology [ 36 ]. The reaction is based on oxidants in the sample being able to oxidize Fe 2+ -o-dianisidine complex to Fe 3+ .…”

“…The peroxide-activity (POX-Act) assay for this study is based on the measurement of total peroxides using a peroxide–peroxidase reaction with tetramethylbenzidine as the substrate for the chromogenic reaction [ 36 ]. The determination of POX-Act has also been previously used in canine serum [ 36 ]. Results are expressed in µmol/L.…”