Erythrocyte agglutination associated with heparin treatment in three horses

In vitro erythrocyte agglutination developed in 3 hospitalized horses receiving heparin treatment. The agglutination caused artifactual decreases in erythrocyte counts and increases in mean corpuscular volume (MCV) values. Treatment of cell suspensions with trypsin eliminated the agglutination and the changes in erythrocyte count and MCV. Similar abnormalities in erythrocyte counts and MCV have been reported in healthy horses treated with heparin and have been cited as evidence of hemolysis and regenerative anemia.     

Electronic measurement of erythrocyte volume and volume heterogeneity in horses during erythrocyte regeneration associated with experimental anemias

Anemia was induced in three groups of horses by moderate or severe acute hemorrhage, or by acetyl phenylhydrazine-induced hemolysis (Groups I, II, and III, respectively). Serial hemograms were done on a multichannel automated blood cell counter with histogram capability. Changes in hematocrit, mean cell volume, erythrocyte number, red cell distribution width (RDW), and standard deviation of erythrocyte volume were examined over time. Significant increases in mean cell volume were first detectable by days 17, 20, and 14 and reached maximum by days 43, 41, and 29, in Groups I, II, and III, respectively (P less than 0.05). Increased mean cell volume was interpreted as reflecting accelerated erythrocyte regeneration; however, not all horses with accelerated regeneration had changes in mean cell volume. Estimated erythrocyte production rate correlated poorly with hematocrit nadir and change in mean cell volume (r = 0.37 and r = 0.36, respectively, P greater than 0.05). In some horses effective regeneration occurs without development of macrocytosis. Mean cell volume remained increased after other parameters returned to control values, suggesting that mean cell volume values may provide retrospective evidence of altered erythrocyte turnover. Anisocytosis as indicated by significant increases in the standard deviation was greatest during the early part of the regenerative response, reaching maximum values on days 30, 28, and 21 in Groups I, II, and III, respectively, and began to decrease as homogeneous repopulation with macrocytes occurred. Red cell distribution width increased significantly only in severe hemorrhage and hemolysis groups, reaching mean maximum values of 24.3 on day 20 and of 26.4 on day 21 in Groups II and III, respectively (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Erythrogram and red cell distribution width of Equidae with experimentally induced anemia

The erythrogram (erythrocyte histogram) and red cell distribution width (RDW) were evaluated in 5 purebred horses and 1 pony of mixed breeding with experimentally induced anemia. Four horses were studied for 6 weeks after 20% of their estimated blood volume was removed on each of 2 consecutive days (40% total blood loss; acute blood-loss group). Two horses were given acetylphenyl hydrazine IV daily, until acute Heinz body hemolytic anemia was induced; the 2 horses were then evaluated for 6 weeks. One horse and the pony had 20% of their estimated blood volume removed via phlebotomy once each week for 8 weeks to induce iron-deficiency anemia (chronic blood-loss group); the horse had been partially depleted of iron before the study began. Weekly blood samples were examined for changes in the erythrogram, RDW, mean cell volume (MCV), and erythrocyte glucose-6-phosphate dehydrogenase activity. Fourteen days after acute blood loss, mild increases were seen in the MCV, which persisted to day 42. The RDW was increased at day 14 and remained increased until day 42; however, the percentage increase was double that of the MCV at days 14, 21, and 28. Erythrograms had mild extensions of the right slope at days 14 to 28. Mean erythrocyte glucose-6-phosphate dehydrogenase activity increased in all 3 groups, but individual concentrations were erratic. In the 2 horses with acute hemolytic anemia, modest increases of similar magnitude were seen in RDW and MCV.(ABSTRACT TRUNCATED AT 250 WORDS)     

The kinetics of hematopoiesis in the light horse III. The hematological response to hemolytic anemia.

The hematological response to acetylphenylhydrazine hemolytic anemia was studied in three standardbred horses. The lifespan of erythrocytes produced during the most severe phase of the anemia were measured with 75-selenomethionine and found to be 144 days as compared to the 139 day lifespan in response to hemorrhagic anemia or 155 days in normal standardbred horses measured previously using the same technique. The erythrocyte counts returned to initial values in 42 days (37, 34 and 54 days) a mean erythrocyte production of 6.4 times 10-12 erythrocytes/day. The mean hemoglobin production was 0.31 gm/kg body weight/day as compared to 0.11 gm Hb/kg/day previously observed in response to hemorrhagic anemia. The mean increase in erythrocyte mean cell volume was 12 mu-3 during the acute response phase to hemolytic anemia in contrast to the absence of a significant increase in the mean cell volume as previously observed during response to hemorrhagic anemia. Free Heinz bodies separated from erythrocytes during the acute phase could not be differentiated from platelets on the hemocytometer counting chamber with standard techniques.     

Erythrocyte macrocytosis in feline leukemia virus associated anemia

Using erythrocyte volume distribution histograms (erythrograms), erythrocyte macrocytosis and anisocytosis were quantitated in 139 cats tested for feline leukemia virus group-specific antigen. Feline leukemia virus-negative cats with non-regenerative anemia or normal packed cell volumes had normal mean corpuscular volume values. Uninfected cats with regenerative anemia had prominent significantly increased macrocytosis and anisocytosis (p less than 0.01). Ninety percent of 62 feline leukemia virus-positive cats had altered erythrograms. Thirty-three feline leukemia virus-positive cats with non-regenerative anemia had marked macrocytosis. Their mean corpuscular volume values (mean 60 fl +/- 2 fl standard error, reference range of 37-49 fl) were significantly greater than those of feline leukemia virus-negative cats except for those with regenerative anemia. Feline leukemia virus-positive, non-anemic cats had significantly increased mean corpuscular volume values of intermediate magnitude. Nine adult cats experimentally infected with feline leukemia virus developed non-regenerative anemia with significant increases in mean corpuscular volume and anisocytosis. However, the macrocytosis observed in these cats was considerably less than in naturally occurring feline leukemia virus-positive cats with non-regenerative anemia. These observations indicate there are events in the pathogenesis of feline leukemia virus-associated anemia other than simple erythroid hypoplasia. We suggest that hemolysis and erythrocyte regeneration occur before erythroid hypoplasia and may partially account for macrocytosis observed in the face of non-regenerative anemia.     

Equine immune-mediated hemolytic anemia associated with Clostridium perfringens infection

Background: Previous studies have suggested an association between equine immune-mediated hemolytic anemia and clostridial infections or neck abscesses.
Objective: The purpose of this report was to describe and characterize the hematologic abnormalities in a horse with Clostridium -associated immune-mediated hemolytic anemia. We also retrospectively evaluated hematologic abnormalities in 8 horses with clostridial myositis or subcutaneous emphysema.
Methods: A 7-year-old Foreign Warm-Blood gelding was evaluated for anemia and a cervical abscess. CBCs and reticulocyte counts were obtained using an Advia 120 analyzer and evaluation of Wright's-stained smears. All cases of equine Clostridium spp. myositis or subcutaneous emphysema over a 7-year period were identified in a retrospective search of the University of Minnesota Veterinary Teaching Hospital database. Clinical, hematologic, and microbiological findings were recorded.
Results: Clostridium perfringens genotype A was isolated from the neck wound of the gelding. The CBC was characterized by severe regenerative anemia, intravascular hemolysis, and RBC agglutination. A direct Coombs' test was positive. Moderate numbers of spheroechinocytes were observed. The total automated reticulocyte count was 4.5% (56,700/μL), with most reticulocytes having low absorbance (mature reticulocytes). The anemia responded to penicillin and steroidal and nonsteroidal immunosuppressive drugs. Of 8 horses with myositis, all of which involved the cervical region, 5 were anemic, 1 had a positive direct Coombs' test, and 2 had increased numbers of type III echinocytes and spheroechinocytes.
Conclusions: The presence of type III echinocytes or spheroechinocytes may be helpful in diagnosing immune-mediated hemolytic anemia associated with clostridial infections in horses. Automated reticulocyte counts may detect very low levels of reticulocytosis in hemolytic anemia in horses.     

The kinetics of hematopoiesis in the light horse II. The hematological response to hemorrhagic anemia.

Hemorrhagic anemia was experimentally produced in three Standardbred horses by removing approximately 63% of the red cell mass and the accompanying plasma during a three day interval. Red cell parameters were examined daily for 45 days and then weekly until termination of the experiment 250 days after production of the anemia. Leukocytes, platelets and bone marrow aspirates were examined at regular intervals for 25 days after the final phlebotomy. At 24 hours after the last bleeding, 75-selenomethionine was injected intravenously to measure the lifespan of the newly produced erythrocytes. The erythrocyte lifespan was found to be 139 days as compared to the 155 day erythrocyte lifespan for three normal standardbred horses measured previously by similar techniques. The maximum decrease in erythrocyte numbers occurred four, two and two days following the last phlebotomy to 43, 39 and 44% of the original values. The prebleeding erythrocyte levels were regained at approximately 63, 91 and 98 days respectively. During the initial 45 days post phlebotomy the maximum increase in mean cell colume was 2, 4 and 7 mj-3 respectively. During the recovery period there was erythrocyte production of 6.84, 6.99, and 6.12 x 10-9 cells/kg/day. At the same time the absolute production of hemoglobin was 44.6, 50.0, and 51.0/gm/day or on a relative basis 0.096, 0.114 and 0.113 gm/kg/day.     

Reduction of CD4+ and CD8+ T lymphocytes during febrile periods in horses experimentally infected with equine infectious anemia virus

Three horses were experimentally infected with equine infectious anemia virus (EIAV). All horses were febrile after inoculation with EIAV and then developed chronic symptoms with intermittent fever. The febrile period was characterized by a rise in body temperature with reduced PBL and erythrocyte counts. Flow cytometric analysis showed that the reduced number of lymphocytes was due to significant decreases in CD4+ and CD8+ T cells in the absence of any change in B cell number. At the end of the febrile period the body temperature began to recover and numbers of CD4+ and CD8+ T cells showed a tendency to increase. For CD8+ T cells, this increase continued for several days after the febrile period. B cell number also significantly increased after the febrile period in two out of three horses. The decrease of CD8+ T cells was greater than that of CD4+ T cells. Although the PBL numbers and the CD4/CD8 ratio returned to the level of the preinoculation period, erythrocyte numbers decreased as the body temperature normalized after each intermittent fever. These results suggest that the recurring cycle of fever accompanied with viremia is caused by a reciprocal relationship between EIAV replication and the host immune response. Furthermore, we demonstrate that the lymphocytic response mitigates fever and viremia in EIAV infection despite the absence of virus neutralizing antibody.     

Studies on some paraclinical indices on intoxication in horses from freshly cut Jimson weed (Datura stramonium)-contaminated maize intended for ensiling

Monitoring of changes in some blood laboratory parameters in 34 horses after ingesting freshly harvested maize that was to be used for ensiling, heavily contaminated with young Datura stramonium plants, is described. For a 7-day period the following parameters were monitored: haemoglobin content (HGB), red blood cell counts (RBC), white blood cell counts (WBC), haematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), differential white cell counts (DWC), erythrocyte sedimentation rate (ESR), protein fractions, aspartate aminotransferase (AST), lactate dehydrogenase (LDH), total bilirubin (TB), direct bilirubin (DB), blood glucose (Glu), total protein (TP), globulin (Glob) and albumin (Alb). The intoxication was accompanied by erythrocytosis, leukocytosis, regenerative left shift neutrophilia, lymphopaenia, eosinopaenia, increased haematocrit values, low erythrocyte sedimentation rate, hyperglycaemia, bilirubinaemia, hypoproteinaemia and increased activity of AST and LDH. No changes occurred in the mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), albumin, globulin and globulin fractions (alpha 1, alpha 2, beta 1, beta 2 and gamma). The blood parameters returned to normal between post-intoxication days 2 and 5. The observed changes in clinical chemistry indices could be used in the diagnosis, differential diagnosis and prognosis of Jimson weed intoxication.     

Association of maximum voluntary dietary intake of freeze-dried garlic with Heinz body anemia in horses

OBJECTIVE: To characterize hematologic and clinical consequences of chronic dietary consumption of freeze-dried garlic at maximum voluntary intake in horses. ANIMALS: 4 healthy sex- and age-matched horses. PROCEDURE: An initial garlic dose (0.05 g/kg, twice daily) was fed to 2 horses in a molasses carrier as part of their normal ration and was gradually increased to maximum voluntary intake (0.25 g/kg, twice daily) over 41 days. Dietary supplementation then continued for a total of 71 days. Two control horses were fed molasses with no garlic with their ration. Blood samples were collected weekly and analyzed for hematologic and biochemical changes, including the presence of Heinz bodies. Recovery of affected blood values was followed for 5 weeks after termination of dietary supplementation with garlic. RESULTS: At a daily dose of > 0.2 g/kg, horses fed garlic developed hematologic and biochemical indications of Heinz body anemia, as characterized by increases in Heinz body score (HBS), mean corpuscular volume (MCV), mean corpuscular hemoglobin, platelet count, and serum unconjugated and total bilirubin concentrations and decreases in RBC count, blood hemoglobin concentration, mean corpuscular hemoglobin concentration, and serum haptoglobin concentration. Recovery from anemia was largely complete within 5 weeks after termination of dietary supplementation with garlic. Heinz body score and MCV remained high at the end of the 5-week recovery period. CONCLUSIONS AND CLINICAL RELEVANCE: Horses will voluntarily consume sufficient quantities of garlic to cause Heinz body anemia. The potential for garlic toxicosis exists when horses are chronically fed garlic. Further study is required to determine the safe dietary dose of garlic in horses.     

Clinical and clinicopathologic variables in adult horses receiving blood transfusions: 31 cases (1999-2005)

OBJECTIVE: To determine clinical and clinicopathologic abnormalities in horses administered a blood transfusion and evaluate effects of blood transfusion on these variables. DESIGN: Retrospective case series. ANIMALS: 31 adult horses that received > or = 1 blood transfusion. Procedures-Medical records of horses receiving a blood transfusion were reviewed to obtain clinical findings, laboratory test results before and after transfusion, adjunctive treatments, transfusion type and volume, response to transfusion, results of donor-recipient compatibility testing, adverse reactions, and outcome. RESULTS: 31 horses received 44 transfusions for hemorrhagic anemia (HG; n = 18 horses), hemolytic anemia (HL; 8), or anemia attributable to erythropoietic failure (EF; 5). Tachycardia and tachypnea were detected in 31 of 31 (100%) and 22 of 31 (71%) horses, respectively, before transfusion. The PCV and hemoglobin concentration were less than the reference range in 11 of 18 horses with HG, 8 of 8 horses with HL, and 5 of 5 horses with EF. Hyperlactatemia was detected in 16 of 17 recorded values before transfusion. Heart rate, respiratory rate, and PCV improved after transfusion, with differences among the types of anemia. Seventeen (54%) horses were discharged, 9 (29%) were euthanized, and 5 (16%) died of natural causes. Adverse reactions were evident during 7 of 44 (16%) transfusions, varying from urticarial reactions to anaphylactic shock. CONCLUSIONS AND CLINICAL RELEVANCE: Abnormalities in clinical and clinicopathologic variables differed depending on the type of anemia. Colic, cold extremities, signs of depression, lethargy, tachycardia, tachypnea, low PCV, low hemoglobin concentration, and hyperlactatemia were commonly detected before transfusion and resolved after transfusion.     

Inherited erythrocyte pyruvate kinase deficiency in a beagle dog

A 1-year-old, female Beagle dog with minimal exercise intolerance was found to have a persistent, severe, and highly regenerative anemia, splenomegaly, and progressive osteosclerosis. Despite near-normal in vitro erythrocyte pyruvate kinase (PK) activity, the authors diagnosed PK deficiency by demonstrating a glycolytic block at the PK step, the lack of normal R-type PK isoenzyme, and the presence of M(2)-type PK in the animal's erythrocytes. The dam had half-normal erythrocyte PK activity, which supports an autosomal recessive mode of inheritance. We conclude from our studies that close similarities exist between erythrocyte PK deficiency in Beagle, Basenji, and West Highland White Terrier dogs and that this form of PK deficiency may be more widespread than previously thought.     

Heparin-induced agglutination of erythrocytes in horses

Heparin was administered subcutaneously 2 times a day for 4 days to 5 horses. An additional group of 5 horses was used as time-matched controls. Significant decreases in PCV, erythrocyte count, and hemoglobin concentration were observed during heparin therapy. The mean corpuscular volume (MCV) of the heparin-treated horses increased to a peak value of 66.1 fl on the last day of treatment. Erythrocyte creatine concentration and glucose 6-phosphate dehydrogenase activity increased moderately during the treatment. These data indicated that the rapid, profound increase in MCV during heparin therapy was not primarily a result of release of large immature erythrocytes from the bone marrow. A second experiment was subsequently performed, using 3 horses. These horses were given heparin 2 times a day, as was done in the first experiment. Saline wet mounts of erythrocyte suspensions were examined once a day for the presence of agglutination. Cell suspensions were examined with or without exposure to a dilute trypsin solution, and erythrocyte counts were done on each suspension, using an electronic cell counter. Agglutination of erythrocytes was evident on the first day of treatment and became more pronounced as treatment progressed. Exposure to trypsin solution reversed the agglutination. The apparent erythrocyte count decreased and MCV increased sharply in the samples processed normally, but there was little change in those suspensions exposed to trypsin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hematologic features of iron deficiency anemia in llamas.

Iron deficiency anemia was identified and characterized in three 14 to 29-month-old male llamas (llama Nos. 1-3) from separate herds in Colorado. The identification of iron deficiency anemia was based on hypoferremia (serum iron = 20-60 micrograms/dl), erythrocytic features, and hematologic response to iron therapy. The anemia was moderate and nonregenerative and characterized by erythrocyte hypochromia, microcytosis (mean cell volume = 15-18 fl), and decreased mean corpuscular hemoglobin concentration (36.0-41.0 g/dl). Morphologic features unique to llamas with iron deficiency anemia included irregular distribution of hypochromia within erythrocytes and increased folded cells and dacryocytes. The cause of iron deficiency was not determined. The llamas were treated with various doses and schedules of parenteral iron dextran. Two of the llamas were monitored for up to 14 months after the start of iron therapy and experienced increases in hematocrit and mean cell volume values. In one llama, progressive replacement of microcytic cells with normal cells was visualized on sequential erythrocyte volume distribution histograms following iron therapy.     

Cloning and tissue expression of the equine transferrin receptor

Background: Characterization of anemia in horses presents a challenge, as they do not release reticulocytes into peripheral blood. Transferrin receptor (TfR) expression is highest on erythroid cells in people and rats, and measurement of a soluble serum form (sTfR) is used to quantify erythropoiesis in these species. We hypothesized that equine TfR (eTfR) expression is similar in quantity and distribution to that in these other species and thus has potential for characterization of the regenerative response in anemic horses. Objective: This study was conducted to clone and sequence the eTfR gene and measure expression levels using quantitative real‐time PCR and immunohistochemical (IHC) staining. Methods: Total RNA from equine bone marrow was used to produce cDNA. The eTfR gene was amplified using pooled gene‐specific primers, and PCR products were sequenced. Rapid amplification of cDNA ends was used to obtain the first 22 nucleotides of the coding sequence. Quantitative PCR was performed using eTfR gene‐specific primers, and IHC staining was used to localize TfR protein expression. Results: The deduced amino acid (aa) sequence (767 aa) of the eTfR was 75–83% identical with sequences of the receptor in several other mammals. As in people and rats, eTfR mRNA expression was highest in the bone marrow, and distribution in other tissues was also similar. Conclusion: The eTfR gene is similar to that of other mammals in structure and expression levels. We hypothesize that it is also similar in function and that, following development of an immunoassay, determining sTfR concentrations will be useful for identifying the regenerative response in anemic horses.     

Pathogenesis, laboratory diagnosis, and clinical implications of erythrocyte enzyme deficiencies in dogs, cats, and horses

Deficiencies of enzymes involved in erythrocyte metabolism can have significant effects on erythrocyte function and survival. Animals with pyruvate kinase (PK) or phosphofructokinase (PFK) deficiencies have shortened erythrocyte life spans and regenerative anemia. PK-deficient dogs (but not PK-deficient cats) develop progressive myelofibrosis and osteosclerosis of bone marrow and hemochromatosis and cirrhosis of the liver. PFK-deficient dogs have sporadic episodes of hyperventilation-induced intravascular hemolysis and hemoglobinuria. Cytochrome b5 reductase (Cb5R) deficiency in dogs and cats results in persistent methemoglobinemia and cyanotic mucous membranes. Severe deficiency of glucose-6-phosphate dehydrogenase, the rate-controlling enzyme in the pentose phosphate pathway, resulted in anemia with eccentrocytosis in an American saddlebred colt. Horses with erythrocyte flavin adenine dinucleotide (FAD) deficiency have both eccentrocytosis (attributable to severe deficiency in glutathione reductase activity) and methemoglobinemia (attributable to Cb5R deficiency); the dual enzyme deficiency occurs because FAD is a required cofactor for both enzymes. Erythrocyte enzyme deficiencies do not usually shorten life expectancy, except for PK-deficient dogs and potentially PFK-deficient dogs during a hemolytic crisis. Although enzyme deficiencies are rare causes of anemia and methemoglobinemia, the ability to diagnose deficient animals allows for the possibility of eliminating these undesirable traits in future breeding. DNA-based assays are available for PK and PFK deficiencies; whereas, biochemical tests of enzyme activity are required for other deficiencies. Continued research is needed to document additional enzyme deficiencies that likely occur and to develop additional DNA-based assays to detect heterozygous animals.     

Chronic lead poisoning in horses

Lead acetate was fed to 4 groups of 2 horses each to study chronic lead intoxication. A 5th group of 3 horses was maintained as controls. The leas was fed in capsules, with the minimum dosage of 6.25 mg/kg/day of lead as lead acetate (group I). The dose was increased from group I through group IV in an approximate geometric series, with each group being given about 125% of the dose given the previous group. These doses were given for 105 days, a period designated as phase 1. Since clinical signs were not observed after 105 days, the doses were increased and fed for an additional 190 days (days 106 to 295). This period was designated phase 2. The smallest daily dose in phase 2 was set at about 125% of the largest daily dose in phase 1. The doses in each group was increased by about 125% of that of the previous group, as was done in phase 1. Seven horses died or were euthanatized after 18 to 190 days of phase 2 (123 to 295 days after the 1st dose). One horse in group I did not develop any clinical signs of intoxication. Dose-related responses were unnoticed with doses larger than 15.3 mg/kg/day. All horses given lead had increased blood lead and serum iron concentrations. During phase 2, the hematocrit (erythrocyte volume) and hemoglobin contents were depressed. The lead concentration in kidney, liver, spleen, pancreas, brain, bone, and heart was increased in the treated horses. The dose level required to produce lead intoxication was greater than that reported for cattle and that estimated in epizootiologic studies of horses.     

Physiologic alterations in the horse produced by food and water deprivation during periods of high environmental temperatures

Eight normal horses were held without access to food or water for 72 hours during a period of high environmental temperatures. During this period, the horses had an average weight loss of 51.6 kg (10.7% of body weight). Highly significant (P less than 0.001) decreases in extracellular fluid volume (18.6 L) and plasma volume (5 L) were observed during this period as compared with base-line values. Plasma protein, sodium, chloride, and osmolality progressively increased in response to the dehydration, whereas packed cell volume, plasma potassium, calcium, magnesium, and phosphate were not significantly altered. The dehydration and clinicopathologic alterations produced were similar to those observed in other species in which the principal problem was a water deficit. After the end of the experimental dehydration period, the horses replaced 62% of the weight loss during a 1-hour period with access to water only.     

Effects of warfarin on blood rheology in navicular disease

A preliminary investigation has been undertaken of blood rheology in horses and ponies, its variation in navicular disease and the changes following treatment with warfarin. Erythrocyte flexibility, measured by a centrifuge packing technique, is higher in horses (30 per cent min-1) than in ponies (23.8 per cent min-1). There are corresponding differences in blood viscosity. The high erythrocyte flexibility in horses is caused by an unknown factor present in plasma. The erythrocyte flexibility in horses with navicular disease is even higher, at 38.5 per cent min-1. Treatment with warfarin reduces the flexibility to just below the normal value. A significant fall in plasma viscosity and erythrocyte flexibility was found after treatment of four ponies with 6 mg warfarin daily for one week.     

Influence of exercise on horses' limb volume, measured by an optoelectronic device

This study tested for the first time an optoelectronic device (Perometer*) measuring limb volume of horses. An analysis of its application was performed regarding the influence of different types of physical exercise on the volume of the extremities. 40 horses were divided into four groups often horses, each group being exposed to exercise, differing in kind and duration of lunging, horse-ridden and riderless. The volume measurement was performed by determining the inactive value (I) after a twelfe-hour standing period, the load value (II) immediately after exercise and recovery value (III) one hour after exercise. The results showed a highly significant decrease in limb volume of 5.7% between values (I) and (II), as well as a highly significant increase of 68% between (II) and (III). The volume changes observed in male horses were significantly higher than in female horses. Volume changes in horse-ridden groups were higher than riderless ones, but not significant. There were fewer leg volume changes at low ambient temperatures (1-12 degrees C) compared to high temperatures (23-31 degrees C), but these were not statistically significant. Well-trained horses showed lower volume changes than less trained horses (not statistically significant). The Perometer is a fast and reliable tool for measuring limb volume of horses. The ICC (Interclass Correlation Coefficient) calculated a high measurement reproducibility of 0.996. A variance analysis showed no significant differences between the three repeated leg measurements. Thus, the Perometer can be used for monitoring and quantifying edema and documenting limb volume changes resulting from training, wearing compression stockings or bandages, manual lymph drainage.