Ethylenediaminetetraacetic acid (EDTA) is a chelating agent commonly used in equine medicine for its ability to bind metal ions. In horses, EDTA is primarily utilized as an anticoagulant in blood sample collection, preventing clotting by sequestering calcium ions, which are essential for the coagulation process. Its use is critical in preserving blood samples for laboratory analysis, ensuring accurate hematological and biochemical evaluations. Additionally, EDTA may be used in chelation therapy to manage heavy metal toxicity in horses. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, mechanisms, and implications of EDTA use in equine veterinary practice.
Oddsdóttir C, Jónsdóttir HK, Sturludóttir E, Vilanova XM.Studies have been carried out on the effect of large-volume blood harvesting from horses, but they were performed on nonpregnant horses of various breeds other than Icelandic horses. This study aimed to investigate the effect on the haematological variables indicating erythropoiesis of repeated blood harvesting from pregnant mares. To account for regional variation, two herds of mares were chosen, both kept under free-range conditions. Sequential EDTA blood samples were collected weekly from 160 mares and analysed for haematological variables in an automated analyser. Serum samples from 115 ma...
Rhodes AD, Wilkins PA, McCoy AM.To compare plasma l-lactate (LAC) values between samples collected from jugular and cephalic venipuncture in healthy horses and systemically ill horses. Methods: Prospective, experimental study. Methods: Large animal university teaching hospital. Methods: Thirty healthy adult university-owned horses and 43 client-owned horses presenting to the large animal hospital for elective surgical procedures or for emergent medical evaluation of systemic illness. Methods: Blood samples were collected from the jugular vein (JV) and cephalic vein (CV) and placed in EDTA blood tubes prior to any medical the...
Oddsdóttir C, Jónsdóttir HK, Sturludóttir E.Few studies have been conducted on haematological reference intervals (RIs) in Icelandic horses. Reference intervals have been published for Icelandic horses in Austria and a preliminary study in Iceland compared haematological values in riding horses to published RIs for other breeds as well as Icelandic horses abroad. Haematological parameters can vary greatly due to factors such as breed, gender, age, reproductive status, and training, as well as feeding, prior exercise and management method. Icelandic broodmares are kept on pasture under supervision throughout the year, with haylage provid...
Dorrego A, Camino E, Gago P, Buendia-Andres A, Acurio K, Gonzalez S, de Juan L, Cruz-Lopez F.Equine piroplasmosis (EP) is caused by Theileria equi and Babesia caballi, transmitted by tick vectors. Horses can suffer an acute, subacute, and chronic forms of the disease, with clinical signs such as poor performance, fever, pale mucosal membranes, and jaundice. The diagnosis of EP subclinical cases is complex due to the sensitivity of real-time PCR and the limited parasite load in some carriers, making it challenging to differentiate them from seropositive, PCR negative (S+PCR-) individuals. This study aimed to describe haematological and biochemical changes in asymptomatic EP carriers, E...
Olaciregui M, Luño V, Martí JI, Aramayona J, Gil L.During the freeze-drying procedure, sperm DNA might become damaged by both freezing and drying stresses. Sperm DNA status can be detected using well-established assays; however, most techniques are expensive and involve elaborate protocols and equipment. Indirect assessments can provide alternative strategies. The objective of this study was to compare a simple test of DNA status using Diff-Quik (DQ) with two established procedures: acridine orange test (AOT) and sperm chromatin dispersion (SCD) on freeze-dried (FD) stallion spermatozoa. Ejaculated spermatozoa from three stallions were freeze-...
Xu J, Cai J, Anderson B, Wagner B, Albrecht R, Peek SF, Suresh M, Darien BJ.The recent molecular characterization and sequencing of equine P-selectin (ePsel), and its glycoprotein ligand, P-selectin glycoprotein ligand-1 (PSGL-1), have provided the tools for further investigation into their role in leukocyte trafficking. Here, we report the generation of a genetically engineered chimeric protein (ePsel-IgG) in which the equine P-selectin lectin and epithelial growth factor (EGF) domains were covalently linked to the equine IgG1 heavy chain constant region. The soluble ePsel-IgG was observed to bind to equine monocytes by confocal microscopy and flow cytometry. Further...
Okubo K, Wilawan P, Bork S, Okamura M, Yokoyama N, Igarashi I.Ethylene glycol bis (beta-aminoethylether)-N,N,N,N-tetraacetic acid (EGTA) is a chelating agent capable of binding to positively-charged metal ions, including a calcium-ion (Ca2+). Here, we demonstrated the inhibitory effect of the chemical on the in vitro asexual growth of the equine protozoan parasites, Babesia caballi and Babesia equi. The growth of both B. caballi and B. equi was significantly inhibited in the presence of EGTA (IC50=1.27 and 2.25 mM, respectively). Under microscopical observation, increased percentages of extracellular merozoites in the total parasites were detected in bot...
Estepa JC, Lopez I, Mayer-Valor R, Rodriguez M, Aguilera-Tejero E.The aim of this study was to evaluate the influence of two commonly used anticoagulants (K3EDTA and lithium heparin) on refractometric and spectrophotometric measurement of total protein (TP) concentration in equine peritoneal fluid samples. The influence of a commercial solution of K3EDTA, a solution of K3EDTA in distilled water and lithium heparin on the refractometric and spectrophotometric (biuret) quantification of TP content in peritoneal fluid samples was assessed. Total protein concentration measured by refractometry was consistently overestimated in samples with commercial K3EDTA. The...
Ajloo D, Moosavi-Movahedi AA, Sadeghi M, Gharibi H.Thermal stabilities of chicken, grey lag goose (Anser anser), turkey as avian hemoglobins (Hbs); and human, bovine, sheep and horse as mammalian Hbs in hemolysate form were investigated and compared with oxygen affinities taken from literature. The thermal stability was obtained from thermal profiles using temperature scanning spectrophotometry. The buffer conditions were 50 mM Tris, pH 7.2, and 1 mM EDTA. The average of the inverse temperature transitions, average hydrophobicity, total van der Waals volume, partial molal volume and hydration potential were calculated by computational methods....
León G, Estrada R, Chaves F, Rojas G, Ovadia M, Gutiérrez JM.The ability of the chelating agent CaNa2EDTA to inhibit local tissue damage induced by Bothrops asper venom was studied in mice and in horses used for polyvalent (Crotalinae) antivenom production. CaNa2EDTA was devoid of toxicity when injected i.m. or s.c. inducing only a mild edema. Preincubation of B. asper venom with CaNa2EDTA inhibited hemorrhagic and dermonecrotic activities, but did not reduce edema-forming and myotoxic effects. A group of horses initially immunized with native venoms developed less severe local tissue reactions when injected with booster doses of venom and CaNa2EDTA tha...
Hinchcliff KW, Kociba GJ, Mitten LA.Thrombocytopenia in horses may be idiopathic or secondary to chronic infectious or inflammatory diseases (eg, equine infectious anemia, lymphosarcoma), drug administration, bone marrow depression, myelophthisic disease, or disseminated intravascular coagulation. This report describes EDTA-dependent pseudothrombocytopenia in a horse. Platelet counts for blood containing EDTA were consistently less than reference range, but platelet counts of blood containing heparin were within reference range. When thrombocytopenia is diagnosed in horses without clinical evidence of a bleeding tendency, EDTA-d...
Kodaka T, Hirayama A, Abe M, Miake K.EDTA-insoluble organic structures of the hypercalcified peritubular matrix (PM) in horse dentine were observed by scanning electron microscopy. The PM was enveloped in double cylindrical structures composed of fibrillar sheaths in the inner and outer peripheries. Between the outer fibrillar sheath and intrinsic fibrils of the intertubular matrix, a calcified cementing membrane existed. Within the PM, warped cone-shaped structures of fibrillar sheaths, overlapping at intervals of 4-6 microns and semiconcentrically surrounding the dentinal tubule, extended from the inner fibrillar towards the ou...
Boyer RF, Clark HM, LaRoche AP.The reductive release of ferritin iron by several naturally occurring o-diphenols was studied. The initial rate of iron release was quantified by spectrophotometric measurement of the Fe(ferrozine)3(2+) complex, which absorbs maximally at 562 nm. The initial rate of iron release was dependent upon o-diphenol concentration, but not on the concentration of the chromophoric chelating agent, ferrozine, Stoichiometric measurements resulted in a ratio of 2Fe(II) released per molecule of o-diphenol. The series of o-diphenols studied included, caffeic acid, chlorogenic acid, dihydrocaffeic acid, 3,4-d...
Youngquist RS, Blanchard TL, Lapin D, Klein W.Four groups of five pony mares each were used to determine if the intrauterine infusion of EDTA-Tris solution caused adverse effects on the endometrium. The uteri of mares were infused with either saline or EDTA-Tris solution or biopsied or sham-biopsied without infusion. Acute endometritis developed in one (20%) to three (60%) mares in each group during the seven days following treatment, but there were no differences (P > 0.05) in the incidence of endometritis among the groups. Endometrial fibrosis was not evident in biopsies taken on days 14, 30 and 60 following infusion of saline or EDT...
van Dyk E, Immelman A, van Heerden JS.After isolation of Pseudomonas aeruginosa from endometrial biopsies of 6 mares they were treated with amikacin sulphate. Three were treated by intra-uterine application of the drug, in one the drug was given by intramuscular injection, in another the intravenous route was used while in the last mare simultaneous local and intravenous treatment was applied. An intra-uterine Tris-EDTA instillation preceeded the uterine amikacin instillations to aid in the breakdown of the capsule around the bacterium. Serum concentrations of amikacin were determined after intravenous and intramuscular administra...
Hussein KS, Jones BE.Glutathione peroxidase (GSH-Px) was determined in whole blood from cows, goats and horses using cumenehydroperoxide as substrate. Heparin was found to be the most suitable anticoagulant. The highest activities of GSH-Px were found with high concentrations of heparin in the blood samples (1000 and 1250 IU/ml of blood). Sodium fluoride and especially EDTA and sodium citrate gave lower activities of the enzyme. Storage of the blood samples at room temperature (~20°C) or in a refrigerator (~5°C) for 3 days resulted in significantly lower activities of the enzyme, especially in horse blood. Gluta...
Crichton RR, Roman F, Roland F.The release of iron from horse spleen ferritin by the chelating agents desferrioxamine B, rhodotorulic acid, 2,3-dihydroxybenzoate, 2,2'-bipyridyl and pyridine-2-aldehyde-2-pyridyl hydrazone (Paphy) has been studied in vitro. Ferritin prepared by classical procedures involving thermal denaturation releases its iron less effectively than ferritin isolated by a modified procedure that avoids this step. Desferrioxamine B and rhodotorulic acid are the most effective in releasing iron from both preparations of ferritin. When FMN is added, iron release by desferrioxamine B, rhodotorulic acid, and 2,...
Boyer RF, Clark HM, LaRoche AP.The reductive release of ferritin iron by several naturally occurring o-diphenols was studied. The initial rate of iron release was quantified by spectrophotometric measurement of the Fe(ferrozine)3(2+) complex, which absorbs maximally at 562 nm. The initial rate of iron release was dependent upon o-diphenol concentration, but not on the concentration of the chromophoric chelating agent, ferrozine, Stoichiometric measurements resulted in a ratio of 2Fe(II) released per molecule of o-diphenol. The series of o-diphenols studied included, caffeic acid, chlorogenic acid, dihydrocaffeic acid, 3,4-d...
Crichton RR, Roman F, Roland F.The release of iron from horse spleen ferritin by the chelating agents desferrioxamine B, rhodotorulic acid, 2,3-dihydroxybenzoate, 2,2'-bipyridyl and pyridine-2-aldehyde-2-pyridyl hydrazone (Paphy) has been studied in vitro. Ferritin prepared by classical procedures involving thermal denaturation releases its iron less effectively than ferritin isolated by a modified procedure that avoids this step. Desferrioxamine B and rhodotorulic acid are the most effective in releasing iron from both preparations of ferritin. When FMN is added, iron release by desferrioxamine B, rhodotorulic acid, and 2,...
Ajloo D, Moosavi-Movahedi AA, Sadeghi M, Gharibi H.Thermal stabilities of chicken, grey lag goose (Anser anser), turkey as avian hemoglobins (Hbs); and human, bovine, sheep and horse as mammalian Hbs in hemolysate form were investigated and compared with oxygen affinities taken from literature. The thermal stability was obtained from thermal profiles using temperature scanning spectrophotometry. The buffer conditions were 50 mM Tris, pH 7.2, and 1 mM EDTA. The average of the inverse temperature transitions, average hydrophobicity, total van der Waals volume, partial molal volume and hydration potential were calculated by computational methods....
Olaciregui M, Luño V, Martí JI, Aramayona J, Gil L.During the freeze-drying procedure, sperm DNA might become damaged by both freezing and drying stresses. Sperm DNA status can be detected using well-established assays; however, most techniques are expensive and involve elaborate protocols and equipment. Indirect assessments can provide alternative strategies. The objective of this study was to compare a simple test of DNA status using Diff-Quik (DQ) with two established procedures: acridine orange test (AOT) and sperm chromatin dispersion (SCD) on freeze-dried (FD) stallion spermatozoa. Ejaculated spermatozoa from three stallions were freeze-...
León G, Estrada R, Chaves F, Rojas G, Ovadia M, Gutiérrez JM.The ability of the chelating agent CaNa2EDTA to inhibit local tissue damage induced by Bothrops asper venom was studied in mice and in horses used for polyvalent (Crotalinae) antivenom production. CaNa2EDTA was devoid of toxicity when injected i.m. or s.c. inducing only a mild edema. Preincubation of B. asper venom with CaNa2EDTA inhibited hemorrhagic and dermonecrotic activities, but did not reduce edema-forming and myotoxic effects. A group of horses initially immunized with native venoms developed less severe local tissue reactions when injected with booster doses of venom and CaNa2EDTA tha...
Okubo K, Wilawan P, Bork S, Okamura M, Yokoyama N, Igarashi I.Ethylene glycol bis (beta-aminoethylether)-N,N,N,N-tetraacetic acid (EGTA) is a chelating agent capable of binding to positively-charged metal ions, including a calcium-ion (Ca2+). Here, we demonstrated the inhibitory effect of the chemical on the in vitro asexual growth of the equine protozoan parasites, Babesia caballi and Babesia equi. The growth of both B. caballi and B. equi was significantly inhibited in the presence of EGTA (IC50=1.27 and 2.25 mM, respectively). Under microscopical observation, increased percentages of extracellular merozoites in the total parasites were detected in bot...
Hussein KS, Jones BE.Glutathione peroxidase (GSH-Px) was determined in whole blood from cows, goats and horses using cumenehydroperoxide as substrate. Heparin was found to be the most suitable anticoagulant. The highest activities of GSH-Px were found with high concentrations of heparin in the blood samples (1000 and 1250 IU/ml of blood). Sodium fluoride and especially EDTA and sodium citrate gave lower activities of the enzyme. Storage of the blood samples at room temperature (~20°C) or in a refrigerator (~5°C) for 3 days resulted in significantly lower activities of the enzyme, especially in horse blood. Gluta...
Hinchcliff KW, Kociba GJ, Mitten LA.Thrombocytopenia in horses may be idiopathic or secondary to chronic infectious or inflammatory diseases (eg, equine infectious anemia, lymphosarcoma), drug administration, bone marrow depression, myelophthisic disease, or disseminated intravascular coagulation. This report describes EDTA-dependent pseudothrombocytopenia in a horse. Platelet counts for blood containing EDTA were consistently less than reference range, but platelet counts of blood containing heparin were within reference range. When thrombocytopenia is diagnosed in horses without clinical evidence of a bleeding tendency, EDTA-d...
Youngquist RS, Blanchard TL, Lapin D, Klein W.Four groups of five pony mares each were used to determine if the intrauterine infusion of EDTA-Tris solution caused adverse effects on the endometrium. The uteri of mares were infused with either saline or EDTA-Tris solution or biopsied or sham-biopsied without infusion. Acute endometritis developed in one (20%) to three (60%) mares in each group during the seven days following treatment, but there were no differences (P > 0.05) in the incidence of endometritis among the groups. Endometrial fibrosis was not evident in biopsies taken on days 14, 30 and 60 following infusion of saline or EDT...
Xu J, Cai J, Anderson B, Wagner B, Albrecht R, Peek SF, Suresh M, Darien BJ.The recent molecular characterization and sequencing of equine P-selectin (ePsel), and its glycoprotein ligand, P-selectin glycoprotein ligand-1 (PSGL-1), have provided the tools for further investigation into their role in leukocyte trafficking. Here, we report the generation of a genetically engineered chimeric protein (ePsel-IgG) in which the equine P-selectin lectin and epithelial growth factor (EGF) domains were covalently linked to the equine IgG1 heavy chain constant region. The soluble ePsel-IgG was observed to bind to equine monocytes by confocal microscopy and flow cytometry. Further...
Estepa JC, Lopez I, Mayer-Valor R, Rodriguez M, Aguilera-Tejero E.The aim of this study was to evaluate the influence of two commonly used anticoagulants (K3EDTA and lithium heparin) on refractometric and spectrophotometric measurement of total protein (TP) concentration in equine peritoneal fluid samples. The influence of a commercial solution of K3EDTA, a solution of K3EDTA in distilled water and lithium heparin on the refractometric and spectrophotometric (biuret) quantification of TP content in peritoneal fluid samples was assessed. Total protein concentration measured by refractometry was consistently overestimated in samples with commercial K3EDTA. The...
Kodaka T, Hirayama A, Abe M, Miake K.EDTA-insoluble organic structures of the hypercalcified peritubular matrix (PM) in horse dentine were observed by scanning electron microscopy. The PM was enveloped in double cylindrical structures composed of fibrillar sheaths in the inner and outer peripheries. Between the outer fibrillar sheath and intrinsic fibrils of the intertubular matrix, a calcified cementing membrane existed. Within the PM, warped cone-shaped structures of fibrillar sheaths, overlapping at intervals of 4-6 microns and semiconcentrically surrounding the dentinal tubule, extended from the inner fibrillar towards the ou...
van Dyk E, Immelman A, van Heerden JS.After isolation of Pseudomonas aeruginosa from endometrial biopsies of 6 mares they were treated with amikacin sulphate. Three were treated by intra-uterine application of the drug, in one the drug was given by intramuscular injection, in another the intravenous route was used while in the last mare simultaneous local and intravenous treatment was applied. An intra-uterine Tris-EDTA instillation preceeded the uterine amikacin instillations to aid in the breakdown of the capsule around the bacterium. Serum concentrations of amikacin were determined after intravenous and intramuscular administra...
Dorrego A, Camino E, Gago P, Buendia-Andres A, Acurio K, Gonzalez S, de Juan L, Cruz-Lopez F.Equine piroplasmosis (EP) is caused by Theileria equi and Babesia caballi, transmitted by tick vectors. Horses can suffer an acute, subacute, and chronic forms of the disease, with clinical signs such as poor performance, fever, pale mucosal membranes, and jaundice. The diagnosis of EP subclinical cases is complex due to the sensitivity of real-time PCR and the limited parasite load in some carriers, making it challenging to differentiate them from seropositive, PCR negative (S+PCR-) individuals. This study aimed to describe haematological and biochemical changes in asymptomatic EP carriers, E...
Oddsdóttir C, Jónsdóttir HK, Sturludóttir E.Few studies have been conducted on haematological reference intervals (RIs) in Icelandic horses. Reference intervals have been published for Icelandic horses in Austria and a preliminary study in Iceland compared haematological values in riding horses to published RIs for other breeds as well as Icelandic horses abroad. Haematological parameters can vary greatly due to factors such as breed, gender, age, reproductive status, and training, as well as feeding, prior exercise and management method. Icelandic broodmares are kept on pasture under supervision throughout the year, with haylage provid...
Rhodes AD, Wilkins PA, McCoy AM.To compare plasma l-lactate (LAC) values between samples collected from jugular and cephalic venipuncture in healthy horses and systemically ill horses. Methods: Prospective, experimental study. Methods: Large animal university teaching hospital. Methods: Thirty healthy adult university-owned horses and 43 client-owned horses presenting to the large animal hospital for elective surgical procedures or for emergent medical evaluation of systemic illness. Methods: Blood samples were collected from the jugular vein (JV) and cephalic vein (CV) and placed in EDTA blood tubes prior to any medical the...
Oddsdóttir C, Jónsdóttir HK, Sturludóttir E, Vilanova XM.Studies have been carried out on the effect of large-volume blood harvesting from horses, but they were performed on nonpregnant horses of various breeds other than Icelandic horses. This study aimed to investigate the effect on the haematological variables indicating erythropoiesis of repeated blood harvesting from pregnant mares. To account for regional variation, two herds of mares were chosen, both kept under free-range conditions. Sequential EDTA blood samples were collected weekly from 160 mares and analysed for haematological variables in an automated analyser. Serum samples from 115 ma...
