Hemolysis in horses refers to the breakdown of red blood cells, which can occur due to various physiological or pathological conditions. This process results in the release of hemoglobin into the bloodstream, potentially leading to anemia and other health issues. Hemolysis can be triggered by factors such as infectious diseases, immune-mediated disorders, toxins, or mechanical damage to red blood cells. The diagnosis and management of hemolysis involve identifying the underlying cause and assessing the extent of red blood cell destruction. This page compiles peer-reviewed research studies and scholarly articles that explore the mechanisms, causes, and clinical implications of hemolysis in equine health.
Pinnell EF, Her J, Gordon D, Kinsella HM, Langston CE, Toribio RE.An adult American Quarter Horse mare presented for pigmenturia and lethargy of 12 hours' duration and was diagnosed with silver maple leaf toxicity. The mare had intravascular hemolysis and azotemia. The mare was treated with a transfusion of whole blood, fluids administered IV, antibiotics, oxygen insufflation, and supportive care. The azotemia persisted despite conventional medical management and hemodialysis was elected. After 2 intermittent hemodialysis treatments over 3 days, the azotemia almost resolved, clinical signs improved, and the mare was discharged. The blood urea nitrogen, c...
Bischoff K.Plants in the maple genus, Acer, and pistachio genus, Pistacia, have been reported to cause acute hemolysis in horses. The cause of hemolysis seems to be metabolism of gallic acids to the potent oxidant pyrogallol by enteric bacteria of the horse. Diagnosis is often tentative and circumstantial. Treatment is symptomatic and supportive and can include detoxification, fluid and electrolyte therapy, supplemental oxygen, and pain control. Corticosteroid and antioxidant therapies do not improve prognosis. Prognosis is guarded to poor but horses that survive 6 days postexposure are expected to reco...
Yamaguchi T, Hirakawa R, Ochiai H.Lipid compositions of mammalian erythrocyte membranes are different among species. Therefore, the information on hemolysis from mammalian erythrocytes is useful to understand membrane properties of human erythrocytes. In this work, pressure-induced hemolysis and hypotonic one were examined using erythrocytes of human, sheep, cow, cat, dog, pig, horse, rat, and mouse. Pressure-induced hemolysis was suppressed by membrane sphingomyelin, whereas hypotonic hemolysis decreased upon increment of cell diameter. Mass spectra of erythrocyte membrane lipids demonstrated that sphingomyelin with an acyl c...
Jamieson CA, Baillie SL, Johnson JP.Transfusion medicine is a crucial part of equine intensive and critical care. Blood transfusions can save lives in both acute and chronic cases of anemia, hemorrhage, and hemolysis. It is vital to have a comprehensive theoretical and practical understanding of the techniques, implications, risks, and complications. This review covers the physiology and pathophysiology of conditions requiring transfusion, as well as step by step guidance for practitioners of all experience levels. This review is designed to serve as a practical reference for those who are treating horses in either the field or ...
Rossi G, Richardson A, Jamaludin H, Secombe C.Paraoxonase-1 (PON-1) activity is a new inflammatory and oxidative marker. Technical effects and biological factors could affect the accuracy of PON-1 activity measurement. We investigated the effects of storage at different temperatures, repeated freeze-thaw cycles, interferences from hemolytic, lipemic, and icteric samples, and seasonal effects on PON-1 activity in horses. We evaluated 2 substrates with an automated spectrophotometer. Ten equine serum samples were stored under different conditions. Although storage at room (21°C) or refrigeration (4°C) temperature induced a statistically s...
McGivney BA, Griffin ME, Gough KF, McGivney CL, Browne JA, Hill EW, Katz LM.Circulating miRNAs (ci-miRNAs) are endogenous, non-coding RNAs emerging as potential diagnostic biomarkers. Equine miRNAs have been previously identified including subsets of tissue-specific miRNAs. In order to investigate ci-miRNAs as diagnostic tools, normal patterns of expression for different scenarios including responses to exercise need to be identified. Human studies have demonstrated that many ci-miRNAs are up-regulated following exercise with changes in expression patterns in skeletal muscle. However, technical challenges such as haemolysis impact on accurate plasma ci-miRNA quantific...
Radakovic M, Davitkov D, Borozan S, Stojanovic S, Stevanovic J, Krstic V, Stanimirovic Z.The aim of this study was to determine the concentrations of oxidative stress parameters and DNA damage in horses infected by Theileria equi. Initial screening of 110 horses with duplex PCR enabled the selection of 30 infected horses with T. equi and 30 free of infection (control). Specimens from the 60 horses were further analysed by determining the following oxidative stress parameters: extent of haemolysis (EH), plasma free haemoglobin (PHb), catalase (CAT), Cu,Zn superoxide dismutase (SOD1), paraoxonase (PON1), nitrite (NO), total nitrate and nitrite (NOx), malondialdehyde (MDA) and free t...
Noble GK, Li X, Zhang D, Sillence MN.Tryptophan (TRP) is marketed as a calmative for horses despite reservations about its efficacy. The aim of this study was to measure the effect of oral TRP administration on the reaction speed of horses. Sixty mature horses were used in a two stage randomised, blind, cross-over study, receiving a placebo and an oral dose of TRP (30, 60 or 120 mg/kg body weight), before undergoing a reaction speed test. Blood samples were taken up to 96 h after TRP administration, to identify signs of acute haemolytic anaemia. Plasma TRP concentrations were increased (P <0.001) by the administration of...
Bannai H, Nemoto M, Tsujimura K, Yamanaka T, Kondo T, Matsumura T.Non-specific hemolysis has often been observed during complement-fixation (CF) tests for equine herpesvirus type-1 (EHV-1), even when the sera have virus-specific CF antibodies. This phenomenon has also been reported in CF tests for various infectious diseases of swine. We found that the sera from 22 of 85 field horses (25.9%) showed non-specific hemolysis during conventional CF testing for EHV-1. Because pretreatment of swine sera with potassium periodate (KIO4) improves the CF test for swine influenza, we applied this method to horse sera. As we expected, horse sera treated with KIO4 did not...
Anderson FL, Secombe CJ, Lester GD.A 9-year-old Thoroughbred gelding was presented for swelling over the left neck and inappetence. There was recent history of intramuscular administration of flunixin meglumine into the left neck. On examination, there was evidence of focal myositis, anaemia, haemolysis and pigmenturia. Culture of aspirated fluid from the left side of the neck produced a heavy growth of a Clostridium species. Complications of infection included haemolytic anaemia, hepatopathy, osteitis and transient hypertrophic cardiomyopathy. Treatment included intravenous fluid therapy, antibiotics, anti-inflammatory drugs, ...
Notin C, Vallon L, Desbordes F, Leleu C.Intense physical exercise produces an excess of reactive oxygen species which can disturb the antioxidant/oxidant balance of the horse in training. Several classes of antioxidant dietary compounds have been suggested to provide health benefits and there is evidence that consumption of these products leads to a reduction in the expression of various pro-inflammatory and/or oxidative stress biomarkers. The recent development of a new galenic system allows the oral delivery of the antioxidant enzyme: superoxide dismutase (SOD). This has been developed from a specific melon variety with a particul...
Cywinska A, Szarska E, Kowalska A, Ostaszewski P, Schollenberger A.Exercise-induced intravascular haemolysis and "sport anemia" are widely reported in human sports medicine. It has been recognized also in horses, however, the clinical importance and the onset of this condition seem different than in human. In this study we investigated the episodes of intravascular haemolysis, indicated by the increase in plasma haemoglobin and the decrease in serum haptoglobin levels, after routine training sessions in race horses. Heart rate and changes in haematological parameters confirmed, that the exertion was relatively high. Intravascular haemolysis did not appear in ...
Paltrinieri S, Meazza C, Giordano A, Tunesi C.Thromboelastometry is used for identifying or monitoring coagulation abnormalities. It has been validated in several species but not in horses and the characteristics of the equine thromboelastogram have not yet been detailed. Objective: The purpose of this study was to validate a thromboelastometer to be used with equine blood and to define the normal equine thromboelastogram. Methods: A Rotem-gamma thromboelastometer (Pentapharm GmbH, Munich, Germany) was used on 38 citrated blood samples to investigate native coagulation, the intrinsic and extrinsic pathways, the function of fibrinogen (lar...
Dickinson CE, Gould DH, Davidson AH, Avery PR, Legare ME, Hyatt DR, DebRoy C.A postpartum mare and foal were presented for evaluation of fever and lethargy in the mare. The mare was diagnosed with endometritis and initially responded well to treatment. On the second day of hospitalization, the mare developed renal insufficiency characterized by oliguria, azotemia, hemolysis, and thrombocytopenia. Concurrently, the foal developed rapidly progressive central nervous system signs culminating in refractory seizures. Both animals failed to respond to treatment and were euthanized. Thrombotic microangiopathy involving glomeruli was evident on microscopic examination of the m...
Veit M, Kabatek A, Tielesch C, Hermann A.Equine arteritis virus (EAV), a member of the newly established family Arteriviridae, is a small, positive-stranded RNA virus. It carries two protein complexes in its envelope, gp5/M and the recently described gp2b/gp3/gp4 complex. We report here on several basic features of EAV replication in cell culture and on the protein composition of virus particles. We have also characterized gp2b, gp3, and gp4 expressed using a baculovirus system in insect cells. Finally, we provide evidence that EAV possess hemagglutinating and hemolytic activity. The hemolysis assay might be useful for determining wh...
Moore JN, Norton N, Barton MH, Hurley DJ, Reber AJ, Donovan DC, Vandenplas ML, Parker TS, Levine DM.Endotoxaemia currently is associated with a poor prognosis in horses. The results of recent trials in other species indicate that phospholipid emulsions reduce the deleterious effects of endotoxin (LPS). However, in a previous study in horses, a 2 h infusion of emulsion caused an unacceptable degree of haemolysis. Objective: Rapid administration of a lower total dose of emulsion would reduce the effects of LPS and induce less haemolysis; the emulsion would reduce inflammatory effects of LPS in vitro. Methods: Twelve healthy horses received an i.v. infusion either of saline or a phospholipid em...
Niinistö K, Raekallio M, Sankari S.The study was undertaken to determine how equine red blood cells (RBCs) survive in storage bags designed for use with human RBCs. Separated RBCs were stored in a routine manner for 35 days and examined every 7 days for storage lesions. Measured parameters included haematology, haemolysis, pH, potassium, lactate, adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG). All tests were performed in vitro. Haematology did not change significantly. Haemolysis increased during storage but did not exceed human limits. pH and 2,3-DPG decreased, while lactate, potassium and ATP increased. RBC...
Watts TJ, Handy RD.The haemolytic effect of verapamil on red blood cells (RBCs) exposed to varying osmolarity was investigated. The experimental approach used a modified red cell haemolysis assay with concentrations of verapamil ranging from 50-1500 microM compared to drug free controls. The time-course of haemolytic effects was also investigated. We also briefly determined the haemolytic effects of verapamil in Ca2+-free conditions (with added EGTA). In conditions representing decreasing osmolarity (dilution from 140-0 mM NaCl) there was a significant increase in erythrocyte haemolysis that was also dependent o...
Inoue Y, Matsui A, Asai Y, Aoki F, Matsui T, Yano H.We investigated the effect of exercise on iron metabolism in horses. Four horses were walked on a mechanical walker for 1 wk (pre-exercise). They then performed moderate exercise on a high-speed treadmill in the first week of the exercise and relative high in the second week and high in the third week. Serum iron was significantly lower in the third week of exercise than in the pre-exercise. Transferrin saturation (TS) was significantly lower in the first and third weeks of exercise than in the pre-exercise. Serum haptoglobin was significantly lower in the first week of exercise than in the pr...
Norman TE, Chaffin MK, Johnson MC, Spangler EA, Weeks BR, Knight R.Five horses were evaluated because of severe cutaneous burn injuries following a barn fire. Gross hemolysis and morphologic changes in RBCs consistent with oxidative damage were detected in all of the horses. Of these horses, 4 became azotemic. The overall goals of treatment included wound care, correction of dehydration and provision of diuresis, control of inflammation, pain management, and prophylaxis against sepsis. After treatment, 2 horses survived and were discharged from the hospital. Red blood cell damage and hemolysis following cutaneous burn injury have been investigated in other sp...
The Journal of nutritionJuly 31, 2004
Volume 134, Issue 8 Suppl 2133S-2140S doi: 10.1093/jn/134.8.2133S
Marlin DJ, Johnson L, Kingston DA, Smith NC, Deaton CM, Mann S, Heaton P, Van Vugt F, Saunders K, Kydd J, Harris PA.Oxidative stress occurs when antioxidant defense mechanisms are overwhelmed by free radicals and may lead to DNA damage, which has been implicated in processes such as aging and diseases such as cancer. The two main techniques presently used to quantify DNA damage are measurement of 8-hydroxydeoxyguanosine and the Comet assay (also known as single-cell gel electrophoresis). The aim of this study was to apply the comet assay to equine peripheral blood mononuclear cells (PBMCs) and identify two conditions in which we hypothesized that oxidative DNA damage would be increased in PBMCs: aging and e...
Ochi S, Oda M, Nagahama M, Sakurai J.Clostridium perfringens alpha-toxin is able to lyse various erythrocytes. Exposure of horse erythrocytes to alpha-toxin simultaneously induced hot-cold hemolysis and stimulated production of diacylglycerol and phosphorylcholine. When A23187-treated erythrocytes were treated with the toxin, these events were dependent on the concentration of extracellular Ca2+ . Incubation with the toxin of BAPTA-AM-treated horse erythrocytes caused no hemolysis or production of phosphorylcholine, but that of the BAPTA-treated erythrocytes did. When Quin 2-AM-treated erythrocytes were incubated with the toxin i...
Boyer JD, Breeden DC, Brown DL.To identify compounds in Acer rubrum that cause hemolysis or oxidation of equine erythrocytes and determine whether these toxins are found in other Acer spp. Methods: Equine erythrocytes. Methods: Washed erythrocytes were incubated with extracts and fractions of Acer spp that were separated by thin layer chromatography. Methemoglobin and hemolysis were measured spectrophotometrically. Compounds within Acer spp fractions associated with cell oxidation or hemolysis were identified by gas chromatography-mass spectrometry. Results: Erythrocytes incubated separately with either A. rubrum, A. saccha...
Guina T, Oliver DB.We cloned the gene encoding a membrane-interactive protein of Borrelia burgdorferi by means of its haemolytic activity in Escherichia coli. The haemolytic activity was erythrocyte-species specific, with progressively decreasing activity for erythrocytes from horse, sheep, and rabbit, respectively. Genetic analysis of the haemolytic determinant revealed two borrelia haemolysin genes, blyA and blyB, that are part of a predicted four-gene operon which is present in multiple copies on the 30 kb circular plasmid(s) of B. burgdorferi B31. blyA encodes a predicted alpha-helical 7.4 kDa protein with a...
Miyoshi S, Sasahara K, Akamatsu S, Rahman MM, Katsu T, Tomochika K, Shinoda S.Vibrio mimicus is a causative agent of human gastroenteritis. This pathogen secretes a pore-forming toxin, V. mimicus hemolysin (VMH), which causes hemolysis by three sequential steps: binding to an erythrocyte membrane, formation of a transmembrane pore, and disruption of the cell membrane. VMH with a molecular mass of 63 kDa was purified by ammonium sulfate precipitation and column chromatography with phenyl Sepharose HP and Superose 6 HR. The hemolytic reaction induced by VMH continued up to disruption of all erythrocytes in the assay system. Moreover, VMH that bound preliminarily to erythr...
Weidmeyer CE, Solter PF.The Incstar(R) SPQ II human haptoglobin (Hpt) (Incstar Corporation, Stillwater, MN) immunoturbidimetric assay was validated for the determination of serum and plasma Hpt concentrations in dogs and horses. The anti-human Hpt antiserum supplied with the assay, displayed monospecificity to both dog and horse serum Hpt by immunoelectrophoresis and Western blotting techniques. The automated immunoturbidimetric assay results correlated well with the cyanmethemoglobin binding assay (r=0.953 for canine serum and r=0.941 for equine serum), and had excellent precision at both high and low serum Hpt conc...
Avellini L, Silvestrelli M, Gaiti A.Oxidative stress develops when the generation of free radicals exceeds the antioxidant capacity of cells or extracellular fluids. It can also occur as a result of physical exercise, and the pathogenesis of exercise-induced myopathies and haemolysis in horses may be related to changes in lipid peroxidation caused by free radicals. Cells have developed biochemical protection against oxidative stress and, as tissues seem to increase their antioxidant defences under chronic activation, training may be one of the ways of increasing antioxidant defences. Accordingly, we tested some enzymatic antioxi...
Salvioli G, Gaetti E, Panini R, Lugli R, Pradelli JM.To evaluate why hemolysis of red blood cells (RBC) by bile acids varies in different mammalian species, we determined the mean corpuscular volume (MCV), lipid content and the concentrations of the conjugates of deoxycholate and of NaCl inducing 50% hemolysis of RBC from healthy humans, pigs, horses, cows, sheep and jaundiced humans. A volume of 0.05 mL of washed RBC at 1% hematocrit, which has the same lipid content but different phospholipid composition and number of erythrocytes (owing to the variable MCV), was incubated in taurodeoxycholate (TDC) solution (0-5 mM) to determine the TDC conce...
Craig AM, Blythe LL, Rowe KE, Lassen ED, Barrington R, Walker KC.Recent evidence concerning the pathogenesis of equine degenerative myeloencephalopathy indicated that low blood alpha-tocopherol values are a factor in the disease process. Variables that could be introduced by a veterinarian procuring, transporting, or storing samples were evaluated for effects on alpha-tocopherol concentration in equine blood. These variables included temperature; light; exposure to the rubber stopper of the evacuated blood collection tube; hemolysis; duration of freezing time, with and without nitrogen blanketing; and repeated freeze/thaw cycles. It was found that hemolysis...
Miyoshi S, Sasahara K, Akamatsu S, Rahman MM, Katsu T, Tomochika K, Shinoda S.Vibrio mimicus is a causative agent of human gastroenteritis. This pathogen secretes a pore-forming toxin, V. mimicus hemolysin (VMH), which causes hemolysis by three sequential steps: binding to an erythrocyte membrane, formation of a transmembrane pore, and disruption of the cell membrane. VMH with a molecular mass of 63 kDa was purified by ammonium sulfate precipitation and column chromatography with phenyl Sepharose HP and Superose 6 HR. The hemolytic reaction induced by VMH continued up to disruption of all erythrocytes in the assay system. Moreover, VMH that bound preliminarily to erythr...
Ochi S, Oda M, Nagahama M, Sakurai J.Clostridium perfringens alpha-toxin is able to lyse various erythrocytes. Exposure of horse erythrocytes to alpha-toxin simultaneously induced hot-cold hemolysis and stimulated production of diacylglycerol and phosphorylcholine. When A23187-treated erythrocytes were treated with the toxin, these events were dependent on the concentration of extracellular Ca2+ . Incubation with the toxin of BAPTA-AM-treated horse erythrocytes caused no hemolysis or production of phosphorylcholine, but that of the BAPTA-treated erythrocytes did. When Quin 2-AM-treated erythrocytes were incubated with the toxin i...
Yoshihara E, Nakae T.In order to develop the cytotoxic liposome, the cytolytic effect of polycationic liposome was examined. Upon incubation of the stearylamine-containing liposome (stearylamine-liposome) with rabbit erythrocyte, a significant extent of hemolysis was observed. Hemolytic activity of the liposome depends on the amount of stearylamine in the liposome membrane. The plots of the initial rate of hemolysis versus the concentration of stearylamine-liposome showed a sigmoidal curve, suggesting that stearylamine-liposomes act cooperatively on the erythrocyte membrane. Hemolytic activity of stearylamine-lipo...
Inoue Y, Matsui A, Asai Y, Aoki F, Matsui T, Yano H.We investigated the effect of exercise on iron metabolism in horses. Four horses were walked on a mechanical walker for 1 wk (pre-exercise). They then performed moderate exercise on a high-speed treadmill in the first week of the exercise and relative high in the second week and high in the third week. Serum iron was significantly lower in the third week of exercise than in the pre-exercise. Transferrin saturation (TS) was significantly lower in the first and third weeks of exercise than in the pre-exercise. Serum haptoglobin was significantly lower in the first week of exercise than in the pr...
Guina T, Oliver DB.We cloned the gene encoding a membrane-interactive protein of Borrelia burgdorferi by means of its haemolytic activity in Escherichia coli. The haemolytic activity was erythrocyte-species specific, with progressively decreasing activity for erythrocytes from horse, sheep, and rabbit, respectively. Genetic analysis of the haemolytic determinant revealed two borrelia haemolysin genes, blyA and blyB, that are part of a predicted four-gene operon which is present in multiple copies on the 30 kb circular plasmid(s) of B. burgdorferi B31. blyA encodes a predicted alpha-helical 7.4 kDa protein with a...
Reimers TJ, Lamb SV, Bartlett SA, Matamoros RA, Cowan RG, Engle JS.Veterinary diagnostic endocrinology laboratories frequently receive hemolyzed plasma, serum, or blood samples for hormone analyses. However, except for the previously reported harm done by hemolysis to canine insulin, effects of hemolysis on quantification of other clinically important hormones are unknown. Therefore, these studies were designed to evaluate effects of hemolysis on radioimmunoassay of thyroxine, 3,5,3'-triiodothyronine, progesterone, testosterone, estradiol, cortisol, and insulin in equine, bovine, and canine plasma. In the first experiment, hormones were measured in plasma obt...
Niinistö K, Raekallio M, Sankari S.The study was undertaken to determine how equine red blood cells (RBCs) survive in storage bags designed for use with human RBCs. Separated RBCs were stored in a routine manner for 35 days and examined every 7 days for storage lesions. Measured parameters included haematology, haemolysis, pH, potassium, lactate, adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG). All tests were performed in vitro. Haematology did not change significantly. Haemolysis increased during storage but did not exceed human limits. pH and 2,3-DPG decreased, while lactate, potassium and ATP increased. RBC...
Cywinska A, Szarska E, Kowalska A, Ostaszewski P, Schollenberger A.Exercise-induced intravascular haemolysis and "sport anemia" are widely reported in human sports medicine. It has been recognized also in horses, however, the clinical importance and the onset of this condition seem different than in human. In this study we investigated the episodes of intravascular haemolysis, indicated by the increase in plasma haemoglobin and the decrease in serum haptoglobin levels, after routine training sessions in race horses. Heart rate and changes in haematological parameters confirmed, that the exertion was relatively high. Intravascular haemolysis did not appear in ...
Avellini L, Silvestrelli M, Gaiti A.Oxidative stress develops when the generation of free radicals exceeds the antioxidant capacity of cells or extracellular fluids. It can also occur as a result of physical exercise, and the pathogenesis of exercise-induced myopathies and haemolysis in horses may be related to changes in lipid peroxidation caused by free radicals. Cells have developed biochemical protection against oxidative stress and, as tissues seem to increase their antioxidant defences under chronic activation, training may be one of the ways of increasing antioxidant defences. Accordingly, we tested some enzymatic antioxi...
Moore JN, Norton N, Barton MH, Hurley DJ, Reber AJ, Donovan DC, Vandenplas ML, Parker TS, Levine DM.Endotoxaemia currently is associated with a poor prognosis in horses. The results of recent trials in other species indicate that phospholipid emulsions reduce the deleterious effects of endotoxin (LPS). However, in a previous study in horses, a 2 h infusion of emulsion caused an unacceptable degree of haemolysis. Objective: Rapid administration of a lower total dose of emulsion would reduce the effects of LPS and induce less haemolysis; the emulsion would reduce inflammatory effects of LPS in vitro. Methods: Twelve healthy horses received an i.v. infusion either of saline or a phospholipid em...
Sentsui H, Kono Y.Horse erythrocytes treated with equine infectious anemia virus hemagglutinin were found to be lysed after incubation with fresh horse serum at 37 degrees C. Fresh guinea pig serum induced more efficient hemolysis than horse serum. Direct immunofluorescence test revealed the adsorption of complement factors on the surface of the erythrocytes. Calcium and magnesium ions were necessary for the hemolysis to take place. Antibody against equine infectious anemia virus enhanced the virus-induced complement-mediated hemolysis. These observations indicated that the classical pathway of complement activ...
The Journal of nutritionJuly 31, 2004
Volume 134, Issue 8 Suppl 2133S-2140S doi: 10.1093/jn/134.8.2133S
Marlin DJ, Johnson L, Kingston DA, Smith NC, Deaton CM, Mann S, Heaton P, Van Vugt F, Saunders K, Kydd J, Harris PA.Oxidative stress occurs when antioxidant defense mechanisms are overwhelmed by free radicals and may lead to DNA damage, which has been implicated in processes such as aging and diseases such as cancer. The two main techniques presently used to quantify DNA damage are measurement of 8-hydroxydeoxyguanosine and the Comet assay (also known as single-cell gel electrophoresis). The aim of this study was to apply the comet assay to equine peripheral blood mononuclear cells (PBMCs) and identify two conditions in which we hypothesized that oxidative DNA damage would be increased in PBMCs: aging and e...
Weidmeyer CE, Solter PF.The Incstar(R) SPQ II human haptoglobin (Hpt) (Incstar Corporation, Stillwater, MN) immunoturbidimetric assay was validated for the determination of serum and plasma Hpt concentrations in dogs and horses. The anti-human Hpt antiserum supplied with the assay, displayed monospecificity to both dog and horse serum Hpt by immunoelectrophoresis and Western blotting techniques. The automated immunoturbidimetric assay results correlated well with the cyanmethemoglobin binding assay (r=0.953 for canine serum and r=0.941 for equine serum), and had excellent precision at both high and low serum Hpt conc...
Yamanaka H, Shimatani S, Tanaka M, Katsu T, Ono B, Shinoda S.The hemolytic activity of Vibrio vulnificus hemolysin (VVH) against erythrocytes from several animal species (sheep, horse, cow, rabbit, chicken) was investigated. VVH was active against erythrocytes from all species, but the amount of VVH causing 50% hemolysis under identical conditions (hemolytic susceptibility to VVH) differed. The degree of 125I-labeled VVH (125I-VVH) binding to each erythrocyte species correlated with the susceptibility of the cells to hemolysis. However, marked differences in the binding ability of 125I-VVH were not observed against liposomes constructed with lipids from...
Paltrinieri S, Meazza C, Giordano A, Tunesi C.Thromboelastometry is used for identifying or monitoring coagulation abnormalities. It has been validated in several species but not in horses and the characteristics of the equine thromboelastogram have not yet been detailed. Objective: The purpose of this study was to validate a thromboelastometer to be used with equine blood and to define the normal equine thromboelastogram. Methods: A Rotem-gamma thromboelastometer (Pentapharm GmbH, Munich, Germany) was used on 38 citrated blood samples to investigate native coagulation, the intrinsic and extrinsic pathways, the function of fibrinogen (lar...
Radakovic M, Davitkov D, Borozan S, Stojanovic S, Stevanovic J, Krstic V, Stanimirovic Z.The aim of this study was to determine the concentrations of oxidative stress parameters and DNA damage in horses infected by Theileria equi. Initial screening of 110 horses with duplex PCR enabled the selection of 30 infected horses with T. equi and 30 free of infection (control). Specimens from the 60 horses were further analysed by determining the following oxidative stress parameters: extent of haemolysis (EH), plasma free haemoglobin (PHb), catalase (CAT), Cu,Zn superoxide dismutase (SOD1), paraoxonase (PON1), nitrite (NO), total nitrate and nitrite (NOx), malondialdehyde (MDA) and free t...
Notin C, Vallon L, Desbordes F, Leleu C.Intense physical exercise produces an excess of reactive oxygen species which can disturb the antioxidant/oxidant balance of the horse in training. Several classes of antioxidant dietary compounds have been suggested to provide health benefits and there is evidence that consumption of these products leads to a reduction in the expression of various pro-inflammatory and/or oxidative stress biomarkers. The recent development of a new galenic system allows the oral delivery of the antioxidant enzyme: superoxide dismutase (SOD). This has been developed from a specific melon variety with a particul...
McGivney BA, Griffin ME, Gough KF, McGivney CL, Browne JA, Hill EW, Katz LM.Circulating miRNAs (ci-miRNAs) are endogenous, non-coding RNAs emerging as potential diagnostic biomarkers. Equine miRNAs have been previously identified including subsets of tissue-specific miRNAs. In order to investigate ci-miRNAs as diagnostic tools, normal patterns of expression for different scenarios including responses to exercise need to be identified. Human studies have demonstrated that many ci-miRNAs are up-regulated following exercise with changes in expression patterns in skeletal muscle. However, technical challenges such as haemolysis impact on accurate plasma ci-miRNA quantific...
Matsuki N, Tamura S, Ono K, Watari T, Goitsuka R, Yamanobe A, Hiraga A, Kubo K, Takagi S, Hasegawa A.To understand the pathogenesis of equine exercise-induced myopathies and hemolysis, changes of phospholipid peroxidation products in the equine middle gluteal muscle and erythrocytes following the high-speed treadmill exercise were studied. In the skeletal muscle, the peroxidized phosphatidylethanolamine (PE) were increased at 24 hours after the exercise. The malondialdehydes (MDAs) were also increased as the protein-bound MDAs following exercise. In the erythrocytes, the peroxidized PE were significantly decreased at 24 hours after the exercise. The protein-bound MDAs were significantly incre...
Klingeborn B, Dinter Z.Antibody to equid herpesvirus 1, which mediates single radial hemolysis, is that responsible for neutralization. Hemagglutination inhibition antibody is not necessarily involved in neutralization or hemolysis.
Anderson FL, Secombe CJ, Lester GD.A 9-year-old Thoroughbred gelding was presented for swelling over the left neck and inappetence. There was recent history of intramuscular administration of flunixin meglumine into the left neck. On examination, there was evidence of focal myositis, anaemia, haemolysis and pigmenturia. Culture of aspirated fluid from the left side of the neck produced a heavy growth of a Clostridium species. Complications of infection included haemolytic anaemia, hepatopathy, osteitis and transient hypertrophic cardiomyopathy. Treatment included intravenous fluid therapy, antibiotics, anti-inflammatory drugs, ...
Grandy JL, McDonell WN.The pH, osmolality, stability, and bacteriostatic characteristics of 5%, 10%, and 15% solutions of guaifenesin were studied. In vitro and in vivo experiments were done to determine the hemolytic potential of the more concentrated solutions on equine blood, as compared with the recommended 5% solution. The primary objective was to determine whether more concentrated solutions could be used clinically. The secondary objective was to determine the optimal diluent (water, 0.9% saline, or 5% dextrose). It was concluded that a 10% solution of guaifenesin made in sterile distilled water was most suit...
Craig AM, Blythe LL, Rowe KE, Lassen ED, Barrington R, Walker KC.Recent evidence concerning the pathogenesis of equine degenerative myeloencephalopathy indicated that low blood alpha-tocopherol values are a factor in the disease process. Variables that could be introduced by a veterinarian procuring, transporting, or storing samples were evaluated for effects on alpha-tocopherol concentration in equine blood. These variables included temperature; light; exposure to the rubber stopper of the evacuated blood collection tube; hemolysis; duration of freezing time, with and without nitrogen blanketing; and repeated freeze/thaw cycles. It was found that hemolysis...
Dickinson CE, Gould DH, Davidson AH, Avery PR, Legare ME, Hyatt DR, DebRoy C.A postpartum mare and foal were presented for evaluation of fever and lethargy in the mare. The mare was diagnosed with endometritis and initially responded well to treatment. On the second day of hospitalization, the mare developed renal insufficiency characterized by oliguria, azotemia, hemolysis, and thrombocytopenia. Concurrently, the foal developed rapidly progressive central nervous system signs culminating in refractory seizures. Both animals failed to respond to treatment and were euthanized. Thrombotic microangiopathy involving glomeruli was evident on microscopic examination of the m...
Salvioli G, Gaetti E, Panini R, Lugli R, Pradelli JM.To evaluate why hemolysis of red blood cells (RBC) by bile acids varies in different mammalian species, we determined the mean corpuscular volume (MCV), lipid content and the concentrations of the conjugates of deoxycholate and of NaCl inducing 50% hemolysis of RBC from healthy humans, pigs, horses, cows, sheep and jaundiced humans. A volume of 0.05 mL of washed RBC at 1% hematocrit, which has the same lipid content but different phospholipid composition and number of erythrocytes (owing to the variable MCV), was incubated in taurodeoxycholate (TDC) solution (0-5 mM) to determine the TDC conce...
Rossi G, Richardson A, Jamaludin H, Secombe C.Paraoxonase-1 (PON-1) activity is a new inflammatory and oxidative marker. Technical effects and biological factors could affect the accuracy of PON-1 activity measurement. We investigated the effects of storage at different temperatures, repeated freeze-thaw cycles, interferences from hemolytic, lipemic, and icteric samples, and seasonal effects on PON-1 activity in horses. We evaluated 2 substrates with an automated spectrophotometer. Ten equine serum samples were stored under different conditions. Although storage at room (21°C) or refrigeration (4°C) temperature induced a statistically s...
Noble GK, Li X, Zhang D, Sillence MN.Tryptophan (TRP) is marketed as a calmative for horses despite reservations about its efficacy. The aim of this study was to measure the effect of oral TRP administration on the reaction speed of horses. Sixty mature horses were used in a two stage randomised, blind, cross-over study, receiving a placebo and an oral dose of TRP (30, 60 or 120 mg/kg body weight), before undergoing a reaction speed test. Blood samples were taken up to 96 h after TRP administration, to identify signs of acute haemolytic anaemia. Plasma TRP concentrations were increased (P <0.001) by the administration of...
Boyer JD, Breeden DC, Brown DL.To identify compounds in Acer rubrum that cause hemolysis or oxidation of equine erythrocytes and determine whether these toxins are found in other Acer spp. Methods: Equine erythrocytes. Methods: Washed erythrocytes were incubated with extracts and fractions of Acer spp that were separated by thin layer chromatography. Methemoglobin and hemolysis were measured spectrophotometrically. Compounds within Acer spp fractions associated with cell oxidation or hemolysis were identified by gas chromatography-mass spectrometry. Results: Erythrocytes incubated separately with either A. rubrum, A. saccha...
Müller F, Segerling M.Complement activity in the serum of eight species has been studied in two ways: by immobilization of sensitized with human or rabbit antibody and by haemolysis of sheep red cells sensitized with rabbit antibody. Serum of the pig, monkey and man was actively haemolytic but contained a heatlabile factor that immobilized unsensitized in the presence of guinea-pig complement and precluded the detection of immune immobilizing activity. Sera of other species, although without action on unsensitized treponemes, even with added guinea-pig complement, differed in their relative haemolytic and immobil...
