Penicillin-induced immune-mediated hemolytic anemia in a horse.

A 5-year-old female American Quarter Horse was determined to have immune-mediated hemolytic anemia after detection of a positive response to a direct Coombs' test. Penicillin-induced immune-mediated hemolytic anemia was confirmed via a direct antiglobulin test, using penicillin-coated RBC. The horse was clinically improved and the anemia resolved in response to supportive care and discontinuation of penicillin treatment. Penicillin should be considered a possible cause of immune-mediated hemolytic anemia in horses.     

Isotype-specific antibodies in horses and dogs with immune-mediated hemolytic anemia

Classes of antibody bound to erythrocytes were determined using direct immunofluorescence (DIF) flow cytometry in 3 horses and 12 dogs with immune-mediated hemolytic anemia (IMHA). Background levels of antibody binding were determined in samples from 12 horses and 12 dogs that were free of clinical disease. The range of nonspecific binding of a fluorescein isothiocyanate (FITC)-conjugated goat anti-equine immunoglobulin G (IgG) was 19.9–36.7%, but was eliminated by the use of the F(ab)₂ fragment of FITC-conjugated goat anti-equine IgG. Background binding by other class-specific antibodies to equine and canine erythrocytes was negligible. The DIF results were compared to the direct antiglobulin (Coombs) test in 5 horses and 20 dogs with anemia. The former assay was more sensitive in dogs with IMHA than was the Coombs' test (100% versus 58%). In contrast, the Coombs' test had better specificity than the DIF assay (100% versus 87.5%, respectively). Using clinical parameters or response to therapy as the comparison, the positive and negative predictive values for the DIF test were 92% and 100% compared to the values of the Coombs' test of 100% and 62%. The DIF assay detected low levels of cells bound with antibody (<30%) in 5 dogs that were Coombs' test-negative. For both species, performance of the DIF test was independent of the prozone effect. Five dogs with IMHA had IgG and IgM on erythrocytes, 5 had IgG, and 2 had IgM. Three horses had surface-bound IgG, including a horse with suspected penicillin-induced IMHA, a foal with neonatal isoerythrolysis, and a foal with clostridial septicemia. The DIF method was valuable in monitoring the response to therapy in the foal with neonatal isoerythrolysis.     

Immune-mediated hemolytic anemia induced by penicillin in horses

Three horses developed severe, immune-mediated hemolytic anemia after treatment with penicillin. The horses had positive direct antiglobulin (Coombs) tests and high titers of IgG antibody that agglutinated penicillin-coated equine red cells. Two of the horses were tested for antibodies to autologous red cell antigens; autoantibodies were not present. Titers of antipenicillin antibody decreased after penicillin was discontinued but IgG antibody was detectable months after recovery. One of the horses was challenged with penicillin; antibody titer increased slightly, but anemia did not develop. Antipenicillin antibody of the IgM class was present in low titer in 23 (77%) of 30 non-anemic horses tested. Apparently, the horse is similar to man in that penicillin-induced anemia is rare but the percentage of individuals with antipenicillin antibody is high.     

Acute recumbency associated with Anaplasma phagocytophilum infection in a horse

An 11-year-old Hanoverian-cross gelding was evaluated because of acute onset of ataxia, recumbency, and fever. At the stable, this and other horses had recently been infested with ticks. Results of analysis of a sample of CSF were within reference limits, but hematologic abnormalities included lymphopenia, thrombocytopenia, mild anemia, and intracytoplasmic inclusion bodies in neutrophils that were consistent with Anaplasma phagocytophilum (previously Ehrlichia equi). Results of serum biochemical analyses were characteristic of infection and included high, unconjugated bilirubin concentration. Other common causes of recumbency in horses, such as equine protozoal myeloencephalitis, infection with eastern or western equine encephalitis viruses and equine herpesvirus-1, West Nile viral encephalitis, trauma, and metabolic disease, were ruled out. The horse responded quickly to i.v. administration of oxytetracycline and recovered fully within 6 days.     

Immune-mediated hemolytic anemia associated with trimethoprim-sulphamethoxazole administration in a horse.

A 10-year-old, thoroughbred gelding was administered sulphonamide drugs during surgical treatment of guttural pouch mycosis. The horse became anemic and a diagnosis of immune-mediated hemolytic anemia was made after other causes of anemia had been ruled out. The anemia resolved after the drugs were withdrawn.     

Persistent thrombocytopenia in a case of equine infectious anemia

Persistent thrombocytopenia was detected in a horse with equine infectious anemia (EIA). The thrombocytopenia was considered to be immune-mediated, developing secondary to infection with EIA virus. Epistaxis, petechial hemorrhages, subcutaneous hematomas, and edema resolved after treatment with corticosteroids; however, the owners requested that the mare by euthanatized because of infection with EIA virus. Although clinical signs attributable to immune-mediated thrombocytopenia may resolve with appropriate treatment, horses with immune-mediated thrombocytopenia secondary to EIA have a guarded to grave prognosis, because of the risk of recurrence and transmission of the EIA virus.     

Surface ultrastructure of pyruvate kinase-deficient erythrocytes in the Basenji dog.

The scanning electron microscope was used to study the surface morphologic features of erythrocytes from a Basenji dog with hereditary hemolytic anemia due to a pyruvate kinase-deficient erythrocytes (RBC). Cells from this dog were compared with RBC from normal dogs and from dogs with regenerative anemias unrelated to pyruvate kinase deficiency. Demonstration of spherical RBC with uniform spicules on their surface (spheroechinocytes) may provide a morphologic explanation for the shortened erythrocytic life-span previously reported in congenital hemolytic anemia of Basenji dogs. Spherical, spiculated RBC were not found in blood from normal dogs or from anemic dogs with reticulocytoses. The surface of reticulocytes from all dogs with regenerative anemia was roughened, with pronounced folding of the cell membrane.     

Lymphosarcoma and associated immune-mediated hemolytic anemia and thrombocytopenia in horses

Three horses with equine lymphosarcoma were examined because of clinical signs including chronic weight loss, respiratory distress, peripheral edema, and chronic colic. Clinicopathologic findings included evidence of an immune-mediated hemolytic anemia. Immune-mediated thrombocytopenia also was diagnosed in 1 of the horses and suspected in another. One horse died in spite of treatment, 1 died 5 hours after surgical removal of a tumor encircling the jejunum, and 1 was euthanatized because of deteriorating condition. Necropsy of each horse revealed extensive neoplastic infiltration of peripheral lymph nodes and abdominal or thoracic viscera with neoplastic lymphocytes.     

Immune-mediated hemolytic anemia in a horse

An 18-year-old Quarter Horse gelding was determined to have immune-mediated hemolytic anemia after detection of autoagglutination of RBC spherocytosis as well as a positive direct Coombs test result. A lack of response to treatment with corticosteroids necessiated the administration of cyclophosphamide and azathioprine. The anemia resolved after treatment with chemotherapeutic drugs.     

Validation of the Sysmex XT‐2000iV hematology system for dogs,cats, and horses. I. Erythrocytes,platelets, and total leukocyte counts

Background: The Sysmex XT‐2000iV is a laser‐based, flow cytometric hematology system that has been introduced for use in large and referral veterinary laboratories. Objective: The purpose of this study was to validate the Sysmex XT‐2000iV for counting erythrocytes, reticulocytes, platelets, and total leukocytes in blood from ill dogs, cats, and horses. Methods: Blood samples from diseased animals (133 dogs, 65 cats, and 73 horses) were analyzed with the Sysmex XT‐2000iV and the CELL‐DYN 3500. Manual reticulocyte counts were done on an additional 98 canine and 14 feline samples and manual platelet counts were done on an additional 73 feline and 55 canine samples, and compared with automated Sysmex results. Results: Hemoglobin concentration, RBC counts, and total WBC counts on the Sysmex were highly correlated with those from the CELL‐DYN (r≥0.98). Systematic differences occurred for MCV and HCT. MCHC was poorly correlated in all species (r=0.33–0.67). The Sysmex impedance platelet count in dogs was highly correlated with both the impedance count from the CELL‐DYN (r=0.99) and the optical platelet count from the Sysmex (r=0.98). The Sysmex optical platelet count included large platelets, such that in samples from cats, the results agreed better with manual platelet counts than with impedance platelet counts on the Sysmex. Canine reticulocyte counts on the Sysmex correlated well (r=0.90) with manual reticulocyte counts. Feline reticulocyte counts on the Sysmex correlated well with aggregate (r=0.86) but not punctate (r=0.50) reticulocyte counts. Conclusion: The Sysmex XT‐2000iV performed as well as the CELL‐DYN on blood samples from dogs, cats, and horses with a variety of hematologic abnormalities. In addition, the Sysmex detected large platelets and provided accurate reticulocyte counts.     

Performance of 2 microtiter canine Coombs' tests

BACKGROUND: The Coombs' test, which detects immunoglobulin or complement on RBC surfaces, has long been the standard for laboratory confirmation of immune-mediated hemolytic anemia (IMHA), a common cause of hemolytic anemia in the dog. This test, however, suffers from relatively low sensitivity. Optimization of test sensitivity would lead to fewer discrepancies between laboratory results and clinical diagnoses, and in some cases institution of appropriate therapy in a timely manner. OBJECTIVES: The objectives of this study were to 1) characterize the sensitivity and specificity of 2 canine Coombs' tests for the detection of IMHA, 2) document the efficacy of using multiple antisera dilutions beyond what is directed by manufacturers, and 3) evaluate the necessity of monovalent antisera in the test protocol. METHODS: Sixty-five canine whole-blood samples submitted for Coombs' testing were evaluated. Patients were classified as IMHA positive or negative based on a set of predetermined criteria. IMHA classification was compared to Coombs' test results from 2 different Coombs' tests adapted to a microtiter-plate format. One Coombs' test (VMRD Coombs' test) utilized a single polyvalent antiserum (VMRD, Inc, Pullman, WA, USA), while a second Coombs' test (University of Minnesota [U of MN] Coombs' test) used both polyvalent and monovalent antisera. RESULTS: Sensitivity and specificity were 61% and 100% for the VMRD Coombs' test, and 82% and 95% for the U of MN Coombs' test. The use of multiple antisera dilutions resulted in 6 additional Coombs' positive test results. All positive Coombs' test results were positive by polyvalent antisera. CONCLUSIONS: When used in a microtiter-plate format, the U of MN Coombs' test was a more sensitive test for the detection of IMHA in canine patients when compared to the VMRD Coombs' test. The use of multiple antisera dilutions increased test sensitivity. Sensitivity, however, was not increased by the use of monovalent antisera in the Coombs' test protocol.     

Idiopathic pure red cell aplasia and nonregenerative immune-mediated anemia in dogs: 43 cases (1988-1999)

OBJECTIVE: To examine clinical features, laboratory test results, treatment, and outcome of dogs with pure red cell aplasia (PRCA) and idiopathic nonregenerative immune-mediated anemia (NRIMA). DESIGN: Retrospective study. ANIMALS: 43 dogs with severe nonregenerative anemia. PROCEDURE: Medical records of dogs determined to have PRCA, NRIMA, or ineffective erythropoiesis on the basis of bone marrow analysis between 1988 and 1999 were reviewed. Criteria for inclusion were > or = 5-day history of severe nonregenerative anemia (Hct < 20%; < 60.0 x 10(3) reticulocytes/microliter) with no underlying diseases. Information was retrieved on signalment, clinical signs, laboratory test results, treatment, and outcome. RESULTS: Median age of the dogs was 6.5 years. Spayed females and Labrador Retrievers were significantly overrepresented. Median Hct was 11% with no evidence of regeneration (median, 1.5 x 10(3) reticulocytes/microliter). Direct Coombs' test results were positive in 57% of dogs. Biochemical abnormalities included hyperferremia and high percentage saturation of transferrin. Bone marrow findings ranged from PRCA (5%) to erythroid hyperplasia (55%). Myelofibrosis was common. Dogs were treated with immunosuppressive drugs and the response was complete, partial, and poor in 55, 18, and 27% of the dogs, respectively. Mortality rate was 28%. CONCLUSIONS AND CLINICAL RELEVANCE: An immune-mediated pathogenesis should be considered in dogs with severe, nonregenerative anemia, normal WBC and platelet counts, hyperferremia, mild clinical signs, and no evidence of underlying disease. Bone marrow findings range from the rare PRCA to erythroid hyperplasia. Myelofibrosis is often detected in affected dogs and may prevent bone marrow aspiration.     

Evaluation of equine hemograms using the ADVIA 120 as compared with an impedance counter and manual differential count

BACKGROUND: The ADVIA 120 is an automated laser cell counter widely used in veterinary medicine. Although specific software for equine samples is available and validated, only a few reports have been published comparing the ADVIA 120 with other methods for equine hemogram evaluation. OBJECTIVES: The purpose of this study was to compare the hematologic values and reference intervals obtained on the ADVIA 120 with those obtained on an impedance cell counter and manual differential counts in healthy horses. METHODS: EDTA-anticoagulated blood samples were obtained from 114 clinically healthy horses of various breeds, both sexes, and 2-6 years of age. Samples were stored for up to 12 hours at 4 degrees C and then analyzed on the ADVIA 120 and the Hemat 8. A 100-cell to 200-cell differential leukocyte count was performed by 3 independent observers on May-Grünwald-Giemsa-stained smears. Intra-assay precision of the ADVIA 120 was determined by analyzing 5 replicates each of 10 of the blood samples. RESULTS: Results from the ADVIA were significantly higher than those from the impedance counter for RBC count, total WBC count, hemoglobin concentration, red cell distribution width, MCH, and MCHC, and significantly lower for HCT and platelet count. Significantly higher neutrophil and basophil counts and significantly lower lymphocyte counts were obtained with the ADVIA 120 compared with manual counts. Based on Passing-Bablok regression analysis, RBC and platelet counts were in good agreement between the 2 analyzers; a constant and proportional bias was present for other values. Coefficients of variation for erythrocyte parameters on the ADVIA were <1%, but were higher for platelet (6%), total WBC (2%), differential WBC (4%-30%), and reticulocyte (75%) counts. CONCLUSIONS: Results obtained with equine samples on the ADVIA 120 were comparable with those obtained on an impedance counter; reference intervals differed statistically but overlapped. The ADVIA had poor precision for reticulocyte and differential leukocyte counts such that the latter should always be verified on smears.     

Management of apposing,full‐thickness tracheal perforations in two horses

Two horses, one 15‐year‐old Arabian gelding and one 10‐year‐old Quarter Horse gelding, presented with a history of marked subcutaneous emphysema. The first case exhibited no external wound, although there was a depression noted on the ventral neck. The second case had a laceration on the ventral aspect of the neck over the trachea. Endoscopic examination revealed both horses to have concurrent dorsal and ventral perforations of the trachea. The horses were managed by placing a short, cuffed, J‐shaped tracheostomy tube in the ventral perforation, while the dorsal perforation healed. The dorsal perforation in the first horse was allowed to heal by second intention, whereas sutures were placed in the dorsal perforation in the second case to reduce the healing time. Both horses were maintained on oral antimicrobial and nonsteroidal anti‐inflammatory medications throughout treatment. The dorsal perforation healed after 13 days in the first horse, and 22 days in the second horse. The ventral perforation healed in both horses by second intention following tracheostomy removal, giving a cosmetically acceptable result. In addition to facilitating tracheal healing, the tracheostomy tubes prevented the progression of subcutaneous emphysema, and promoted its resolution.     

Splenectomy as adjunctive therapy for immune-mediated thrombocytopenia and hemolytic anemia in the dog

Splenectomy was done in 9 dogs having immune-mediated hematologic disorders refractory to medical therapy. These disorders were immune-mediated thrombocytopenia (n = 3), immune hemolytic anemia (n = 3), and Evan's syndrome (n = 3). The diagnoses were based on clinical observations, laboratory test data, and differential of other conditions. In the 12 months after splenectomy was done, the dogs reflected clinical improvement and return of platelet and/or erythrocyte counts to clinically acceptable limits; medical treatment was stopped or reduced in 8 of 9 patients. The exceptional patient had shown clinical improvement without change in the platelet count. At the end of 1 year, survival rate was excellent, and postsplenectomy complications, such as hemobartonellosis, did not appear. It is believed that splenectomy may be useful for treating immune-mediated thrombocytopenia, anemia, and Evan's syndrome that seem refractory to medication.     

Propylthiouracil-associated hemolytic anemia, thrombocytopenia, and antinuclear antibodies in cats with hyperthyroidism

Nine of 105 cats with hyperthyroidism treated with propylthiouracil developed a serious immune-mediated drug reaction during treatment. Adverse clinical signs, which developed after 19 to 37 days (mean, 24.8 days) of propylthiouracil administration, included lethargy, weakness, anorexia, and bleeding diathesis. Physical examination revealed pale mucous membranes, and petechial hemorrhages of the skin and oral cavity. Results of hematologic testing revealed severe anemia and thrombocytopenia. The direct antiglobulin (Coombs') test was positive in all 7 cats evaluated, whereas the serum antinuclear antibody titer was greater than or equal to 1:10 in 5 of the 8 cats tested. In 4 of the cats, treatment included appropriate supportive therapy and cessation of propylthiouracil; in these cats, anemia and thrombocytopenia resolved and Coombs' and antinuclear antibody tests became negative within 2 weeks.     

Intra-abdominal hypertension in two adult horses

Objective – To describe the clinical and pathological findings in 2 adult horses with documented increases in intra-abdominal pressure (IAP), and to describe the direct puncture technique used to measure this pressure.
Series Summary – Two adult horses developed increases in IAP secondary to large-volume abdominal effusion. A 9-year-old Quarter Horse cross gelding was presented for evaluation of urinary and neurologic signs. Abdominal ultrasonographic examination showed a hepatic abscess along with abdominal effusion. A 4-year-old Quarter Horse gelding was presented for evaluation of castration complications. A castration site infection extended into surrounding tissues, resulting in peritonitis, abdominal effusion, and severe cellulitis of the limbs. IAP measured in both horses was increased relative to reported equine reference values. Changes in hemodynamic parameters in both horses, notably increased central venous pressure, were consistent with those seen in other species in which intra-abdominal hypertension (IAH) and its sequellae have been documented.
New or Unique Information Provided – Extensive research and clinical literature guides management of humans with IAH and abdominal compartment syndrome. Knowledge of these conditions in companion animal and large domestic species is less well developed. Recent research has established reference values for standing, sedated and recumbent, anesthetized horses. Detailed reports of equine clinical cases of IAH have not been reported in the literature. This report provides information on the clinical, hemodynamic, and pathologic characteristics of 2 horses with measured increases in IAP, and describes the direct puncture technique used to perform these measurements.     

Hematology of healthy Florida manatees (Trichechus manatus)

Background: Hematologic analysis is an important tool in evaluating the general health status of free‐ranging manatees and in the diagnosis and monitoring of rehabilitating animals. Objectives: The purpose of this study was to evaluate diagnostically important hematologic analytes in healthy manatees (Trichechus manatus) and to assess variations with respect to location (free ranging vs captive), age class (small calves, large calves, subadults, and adults), and gender. Methods: Blood was collected from 55 free‐ranging and 63 captive healthy manatees. Most analytes were measured using a CELL‐DYN 3500R; automated reticulocytes were measured with an ADVIA 120. Standard manual methods were used for differential leukocyte counts, reticulocyte and Heinz body counts, and plasma protein and fibrinogen concentrations. Results: Rouleaux, slight polychromasia, stomatocytosis, and low numbers of schistocytes and nucleated RBCs (NRBCs) were seen often in stained blood films. Manual reticulocyte counts were higher than automated reticulocyte counts. Heinz bodies were present in erythrocytes of most manatees. Compared with free‐ranging manatees, captive animals had slightly lower MCV, MCH, and eosinophil counts and slightly higher heterophil and NRBC counts, and fibrinogen concentration. Total leukocyte, heterophil, and monocyte counts tended to be lower in adults than in younger animals. Small calves tended to have higher reticulocyte counts and NRBC counts than older animals. Conclusions: Hematologic findings were generally similar between captive and free‐ranging manatees. Higher manual reticulocyte counts suggest the ADVIA detects only reticulocytes containing large amounts of RNA. Higher reticulocyte and NRBC counts in young calves probably reflect an increased rate of erythropoiesis compared with older animals.     

Pathology of the mononuclear cell leukemia of Fischer rats. II. Hematology

Complete hemograms were evaluated for 57 rats with mononuclear cell leukemia and compared to hemograms obtained from 52 age- and sex-matched nonleukemic rats. All leukemic rats had marked hemolytic anemia and associated spherocytosis, reticulocytosis, anisocytosis, and polychromasia. The anemia varied with the stage of illness and was more severe in rts with advanced leukemia. Death appeared to be related to anemia. There was a marked neutrophilia with left shift, mild lymphopenia, and moderate to severe thrombocytopenia. Atypical mononuclear cells were detected in circulation in all but three rats. Total white blood cell counts ranged from 5.0-370 x 10(3) cells/ml. There was an increase in erythrocyte osmotic fragility with separation into two distinct populations of erythrocytes. Eight of nine rats were Coombs' positive indicating an immune-mediated pathogenesis for the anemia. Hemostasis tests revealed a markedly prolonged prothrombin time, hypofibrinogenemia, slightly increased to normal partial thromboplastin time, and undetected fibrin degradation products. These findings suggest significant liver disease associated with the leukemia.     

Clinical application of reticulocyte counts in dogs and cats.

Reticulocytes are anucleate immature red blood cells that contain a network of RNA, organelles, and mitochondria, which stain with supravital dyes. Both aggregate and punctate reticulocytes are present in domestic cats, and aggregate reticulocytes are used to assess the degree of regeneration in anemic dogs and cats. Multiple factors influence the degree of regenerative response to anemia. These factors include time of reticulocyte measurement, concurrent diseases, species, and ongoing therapy. Although many automated systems for reticulocyte enumeration exist, manual counts remain the gold standard in veterinary medicine.