Use of a direct enzyme-linked antiglobulin test for laboratory diagnosis of immune-mediated hemolytic anemia in dogs.

Detection of autoantibody, complement, or both bound to RBC is an essential requirement for unequivocal diagnosis of immune-mediated hemolytic anemia in dogs. An enzyme-linked antiglobulin test was adapted for laboratory diagnosis of this disease. The refinement and routine use of this assay have allowed further observation of the pathogenesis of the disease process. In particular, degree of hemolysis can be related to the degree of RBC sensitization associated with primary immune-mediated hemolytic anemia, and this correlation is highest for IgG autoantibody. Results indicate that autoantibody isotype might have an important role in the hemolytic process.     

Influence of drug treatment on survival of dogs with immune-mediated hemolytic anemia: 88 cases (1989-1999)

OBJECTIVE: To evaluate association of various treatments for immune-mediated hemolytic anemia with survival to discharge in dogs. DESIGN: Retrospective cross-sectional analysis. ANIMALS: 88 dogs with idiopathic immune-mediated hemolytic anemia. PROCEDURE: Medical records of dogs with immune-mediated hemolytic anemia treated between August 1989 and August 1999 were examined. Survival to discharge, PCV at referral, autoagglutination, and drug treatment and dosage were recorded. RESULTS: Treatments included administration of prednisone, dexamethasone, azathioprine, danazol, cyclosporine, cyclophosphamide, bovine hemoglobin solution, and human immunoglobulin. Overall mortality rate was 50.5%. Significant associations with death were not detected for use of azathioprine, cyclosporine, danazol, or human immunoglobulin. A significant difference in mortality rate was not detected between use of multiple immunosuppressive drug treatments and use of single immunosuppressive drugs. Use of cyclophosphamide and bovine hemoglobin solution were associated with significant increases in relative risk of death CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that use of cyclophosphamide and bovine hemoglobin solution in treatment of idiopathic immune-mediated hemolytic anemia may be associated with increased risk of death.     

Immune-mediated Hemolytic Anemia and Thrombocytopenia in the Dog: A retrospective study of 55 cases diagnosed from 1979 through 1983 at the Western College of Veterinary Medicine

All recognized cases (n = 55) of immune-mediated hemolytic anemia and immune-mediated thrombocytopenia in dogs presented to the Western College of Veterinary Medicine from 1969 through 1983 were reviewed. Specific areas of concern were: association with other conditions, therapeutic response, prognosis, relapse rate and final outcome. Of these 55 cases, 19 were immune-mediated hemolytic anemia, 26 were immune-mediated thrombocytopenia and 10 were both immune-mediated hemolytic anemia and thrombocytopenia. Females were slightly over-represented and the mean age was 6.4 years. Therapy consisted of various combinations of immuno-suppressive drugs and in some cases, whole blood transfusion and splenectomy. No firm conclusions could be made regarding therapeutic efficacy, as a result of variation in treatment protocol and the occasional unavailability of follow-up data. Well over half of all cases were diagnosed as idiopathic. Precipitating factors or diseases most frequently implicated in secondary immune-mediated thrombocytopenia or hemolytic anemia were: recent vaccination, drug therapy, obstetrical complications, stress, recent viral infection and neoplasia. Twice as many cases of immune-mediated hemolytic anemia were seen in the cooler months (October to March), although this could not be related to antibody class or thermal reactivity. Immune-mediated thrombocytopenia both as a single disease and combined with immune-mediated hemolytic anemia had no seasonal incidence. History, clinical findings and hematological and clinical chemistry findings were consistent with data previously reported, with the exception of icterus, which appeared to be of higher incidence than most reports, being present in almost 50% of immune-mediated hemolytic anemia cases. Just over half of all dogs survived, although the survival rate was highest for immune-mediated hemolytic anemia, followed closely by immune-mediated thrombocytopenia and lowest for the combined disease. Immune-mediated thrombocytopenia most frequently ran a relapsing course requiring long-term or intermittent therapy.     

Anemia, splenomegaly, and increased osmotic fragility of erythrocytes in Abyssinian and Somali cats

OBJECTIVE: To determine clinical and clinicopathologic features of a chronic intermittent severe hemolytic anemia characterized by erythrocyte osmotic fragility in Abyssinian and Somali cats. DESIGN: Case series. ANIMALS: 13 Abyssinian and 5 Somali cats. PROCEDURES: History, pedigree information, and results of routine laboratory tests, special erythrocyte studies, and histologic evaluation of splenic and hepatic specimens were analyzed. RESULTS: Age at which clinical signs of anemia were first apparent ranged from 6 months to 5 years. Ten cats had splenomegaly. Most often, the PCV was between 15 and 25%, but it was as low as 5% at some times. The anemia was characterized by macrocytosis and mild to moderate reticulocytosis, but no poikilocytosis. Hyperglobulinemia, lymphocytosis, mild hyperbilirubinemia, and high hepatic enzyme activities were common findings. Results of Coombs tests and tests for infectious diseases were negative. The erythrocytic osmotic fragility was high in affected cats (mean osmotic fragility, 0.66 to 0.78%), compared with healthy cats (0.48 to 0.58). No specific membrane protein abnormality, erythrocyte enzyme deficiency, or hemoglobinopathy was identified. Histologic evaluation of splenic and hepatic specimens revealed extramedullary hematopoiesis and hemosiderosis. Four of the 5 Somali cats were closely related. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of results of pedigree analyses, the apparent breed predilection, and the exclusion of other known causes of anemia in cats, we believe that the hemolytic anemia in these cats was likely a result of a novel hereditary erythrocyte defect. A genetic predisposition to immune-mediated destruction of erythrocytes could not be ruled out.     

Pulmonary thromboembolism in cats: 29 cases (1987-1997)

OBJECTIVE: To determine clinical signs, clinicopathologic abnormalities, radiographic findings, histologic abnormalities, and predisposing factors or diseases in cats with pulmonary thromboembolism (PTE). DESIGN: Retrospective study. ANIMALS: 29 cats in which PTE was confirmed at necropsy. PROCEDURE: Information on signalment, body weight, history, results of physical examination, results of CBC and serum biochemical analyses, whether PTE was suspected prior to death, type of indwelling venous catheter and duration of venous catheterization, results of thoracic radiography, and whether cats had any concurrent diseases was obtained from medical records. RESULTS: PTE was identified in cats of various ages (median, 8.7 years), weights (median, 4.1 kg [9 lb]), and breeds. The most common owner-reported problems included lethargy (17 cats), anorexia (14), weight loss (10), and difficulty breathing (8); physical abnormalities included lethargy (21), tachypnea or dyspnea (16), and dehydration (13). Clinicopathologic abnormalities reflected concurrent or underlying diseases. Common radiographic abnormalities included pulmonary vessel abnormalities (11), pleural effusion (8), and peripheral noncircumscribed consolidations (6). Underlying or predisposing conditions, including cardiac disease (12), neoplasia (10), corticosteroid administration (8), disseminated intravascular coagulation (5), protein-losing nephropathy (4) or enteropathy (4), immune-mediated hemolytic anemia (2), and sepsis (2) were identified in all cats. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that PTE can develop in cats of any age, sex, or breed. Because PTE is a serious, potentially life-threatening disease, it should be suspected in cats with thoracic radiographic changes suggestive of uneven distribution of blood flow between lung lobes, especially in cats that have predisposing factors or diseases.     

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.     

A novel DNA virus associated with feather inclusions in psittacine beak and feather disease

The nature of feather inclusions was characterized in 32 psittacine birds (30 cockatoos, one peach-faced lovebird (Agapornis roseicollis), and one red-lored Amazon parrot (Amazona autumnalis autumnalis] with naturally-acquired psittacine beak and feather disease. Intranuclear inclusions within feather epithelial cells and intracytoplasmic inclusions within macrophages in the feather epithelium and pulp cavity contained psittacine beak and feather disease viral antigen when stained by the avidin-biotin complex immunoperoxidase technique. Ultrastructurally, inclusions were observed primarily within macrophages and to a lesser extent within epithelial cell nuclei. Macrophage inclusions appeared as paracrystalline arrays of viral particles. Intranuclear inclusions were less well defined, although scattered viral particles were present. Intracytoplasmic and intranuclear particles in ultrastructural preparations were identified by colloidal gold labeling as psittacine beak and feather disease virus. Feather epithelium was more frequently and severely involved in the disease process than was adjacent follicular epithelium. Plucked feathers with an intact epidermal collar and feather epithelium were preferred to follicular biopsies for histopathologic examination.     

Clinicopathologic findings in dogs seroreactive to Bartonella henselae antigens

OBJECTIVE: To assess the potential clinical relevance of seroreactivity to Bartonella henselae antigens in dogs. ANIMALS: 40 dogs seroreactive to B henselae and 45 dogs that did not seroreact to B henselae. PROCEDURE: A case-control study was conducted. Clinical and clinicopathologic findings were extracted from medical records of each dog. RESULTS: Statistical differences were not detected between dogs seroreactive or nonseroreactive to B henselae when analyzed on the basis of disease category or results of hematologic, biochemical, urine, or cytologic analysis. However, seroreactivity to B henselae antigens was detected in 2 of 4 dogs with a clinical diagnosis of granulomatous meningoencephalitis, 3 of 4 dogs with immune-mediated hemolytic anemia, 3 of 4 dogs with infective endocarditis, 2 of 3 dogs with lymphoid neoplasia, and 5 of 10 dogs with polyarthritis. Additionally, seroreactivity to B henselae antigens was detected in 18 of 34 thrombocytopenic dogs and 14 of 27 dogs with neutrophilia. CONCLUSIONS AND CLINICAL RELEVANCE: Significant associations were not detected between seroreactivity to B henselae and various diseases. Prospective epidemiologic studies investigating specific diseases, such as meningoencephalitis or polyarthritis, and specific hematologic abnormalities, such as immunemediated hemolytic anemia or thrombocytopenia, should be conducted to further define the potential clinical relevance of antibodies against B henselae in dogs. IMPACT FOR HUMAN MEDICINE: Bartonella organisms are increasingly reported as pathogens that induce are increasingly reported as pathogens that induce chronic infections in humans and dogs. Dogs may serve as natural candidates for future study of the disease in humans.     

Antibody response to and maternal immunity from an experimental psittacine beak and feather disease vaccine.

Adult umbrella cockatoos, Moluccan cockatoos, African grey parrots, and a yellow-headed Amazon parrot were inoculated IM or SC with beta-propiolactone-treated psittacine beak and feather disease (PBFD) virus. Thirty- to 45-day-old African grey parrot, umbrella cockatoo, and sulphur-crested cockatoo chicks also were vaccinated with the same inoculum. The hemagglutination inhibition (HI) and agar-gel diffusion tests were used to assay for post-vaccination development of anti-PBFD virus antibodies. All adult vaccinates seroconverted and had increases in HI and precipitating antibodies. The vaccinated chicks had increased concentrations of HI antibodies, but precipitating antibodies could not be detected. To demonstrate that chicks from vaccinated hens are protected from PBFD virus challenge, 3 African grey parrot chicks and 2 umbrella cockatoo chicks from vaccinated hens and 1 African grey parrot chick and 1 umbrella cockatoo chick from nonvaccinated hens were exposed to purified PBFD virus. Chicks from the vaccinated hens remained clinically normal during the 50-day test period. Chicks from the nonvaccinated hens developed clinical and histologic lesions of PBFD. Infected tissues from these birds were confirmed to contain viral antigen, using immunohistochemical staining techniques. The PBFD virus was recovered from the affected birds. These findings indicate that adult and 30- to 45-day-old psittacine birds will seroconvert following vaccination with beta-propiolactone-treated PBFD virus. Also, hens inoculated with beta-propiolactone-treated PBFD virus produce chicks that are, at least temporarily, resistant to virus challenge.     

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.     

Suspected primary pure red cell aplasia in a 4-month-old intact male mixed breed Bernese mountain dog

A 4-month-old, 31-kg intact male mixed-breed Bernese mountain dog was presented for evaluation of severe non-regenerative anemia after several days of lethargy, inappetence and pale mucous membranes. Bone marrow evaluation and complete response to immunosuppressive therapy were suggestive of primary pure red cell aplasia (PRCA). Primary PRCA is a rare immune-mediated non-regenerative anemia that is overrepresented in middle-aged to older spayed female dogs and has not previously been described in an intact male puppy.     

Underlying medical conditions in cats with presumptive psychogenic alopecia

OBJECTIVE: To identify underlying medical conditions in cats with a presumptive diagnosis of psychogenic alopecia. DESIGN: Case series. ANIMALS: 21 adult cats referred with a presumptive diagnosis of psychogenic alopecia. PROCEDURES: A detailed behavior and dermatologic questionnaire was completed by the primary caregiver, and complete behavioral and dermatologic examinations were performed. A standard diagnostic testing protocol that included cytologic examination of skin scrapings, fungal culture of hairs, evaluation of responses to parasiticides and an exclusion diet, assessment for atopy and endocrinopathies, and histologic examination of skin biopsy specimens was used to establish a definitive diagnosis in all cats. Cats that did not respond to an elimination diet were treated with methylprednisolone acetate to determine whether pruritus was a factor. RESULTS: Medical causes of pruritus were identified in 16 (76%) cats. Only 2 (10%) cats were found to have only psychogenic alopecia, and an additional 3 (14%) cats had a combination of psychogenic alopecia and a medical cause of pruritus. An adverse food reaction was diagnosed in 12 (57%) cats and was suspected in an additional 2. All cats with histologic evidence of inflammation in skin biopsy specimens were determined to have a medical condition, but of 6 cats without histologic abnormalities, 4 had an adverse food reaction, atopy, or a combination of the 2, and only 2 had psychogenic alopecia. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that psychogenic alopecia is overdiagnosed in cats. Thorough diagnostic testing should be done before ascribing a behavioral cause to hair loss in cats.     

Diagnostic value of cytologic examination of gastrointestinal tract tumors in dogs and cats: 83 cases (2001-2004)

OBJECTIVE: To determine results of cytologic examination of fine-needle aspirates and impression smears of gastrointestinal tract tumors in dogs and cats. DESIGN: Retrospective case series. ANIMALS: 38 dogs and 44 cats with histologically confirmed gastrointestinal tract tumors. PROCEDURES: Results of cytologic examination of fine-needle aspirates (n = 67) or impression smears (31) were compared with the histologic diagnosis, and extent of agreement was classified as complete, partial, none, or undetermined. RESULTS: For 48 of the 67 (72%) fine-needle aspirates, there was complete or partial agreement between the cytologic and histologic diagnoses. For 12 (18%) aspirates, the extent of agreement could not be determined because the cytologic specimen was considered unsatisfactory. For 29 of the 31 (94%) impression smears, there was complete agreement between the cytologic and histologic diagnoses, and for 2 (6%), there was partial agreement. None of the impression smears were considered unsatisfactory. Proportion of samples with complete agreement and proportion of samples with complete or partial agreement were significantly higher for impression smears than for fine-needle aspirates. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that there was moderate agreement between results of cytologic examination of fine-needle aspirates from dogs and cats with gastrointestinal tract neoplasia and the definitive histologic diagnosis. The agreement between results of cytologic examination of impression smears and the histologic diagnosis appeared to be higher.     

Cloacal papillomatosis in the absence of herpesvirus and papillomavirus in a sulphur-crested cockatoo (Cacatua galerita)

CASE HISTORY A 21-year-old male sulphur-crested cockatoo (Cacatua galerita) was presented following the sudden appearance of blood associated with the passage of faeces and urates.

CLINICAL AND PATHOLOGICAL FINDINGS: There was fresh blood-staining of the feathers around the vent. The dorsal mucosal wall of the proctodeum was erythematous and roughened in appearance. An endoscopic biopsy was performed, and histological examination revealed multiple fronds of epithelium; the mucosa varied from simple to pseudostratified columnar epithelium, with diffuse hyperplasia of goblet cells. The underlying connective tissue stroma was well vascularised and was infiltrated with mixed inflammatory cells, comprising granulocytic cells and macrophages. PCR testing for both herpesvirus and papillomavirus, using consensus primers, was negative.

DIAGNOSIS: Cloacal papillomatosis.

CLINICAL RELEVANCE: This case manifested typical clinical signs and histological lesions of cloacal papillomatosis in the absence of demonstrable herpesvirus or papillomavirus. Veterinarians need to consider this disease in the differential diagnosis of blood in the droppings of parrots and cockatoos even in countries where psittacine herpesviruses are exotic diseases.     

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.     

Multisystemic infection with an Acanthamoeba sp in a dog

CASE DESCRIPTION-A 10-month-old Boxer was evaluated for fever and signs of cervical pain. CLINICAL FINDINGS-Physical examination revealed lethargy, fever, and mucopurulent ocular and preputial discharge. On neurologic examination, the gait was characterized by a short stride. The dog kept its head flexed and resisted movement of the neck, consistent with cervical pain. Clinicopathologic findings included neutrophilic leukocytosis, a left shift, and monocytosis. Cervical radiographs were unremarkable. Cerebrospinal fluid analysis revealed neutrophilic pleocytosis and high total protein content. On the basis of signalment, history, and clinicopathologic data, a diagnosis of steroid-responsive meningitis-arteritis was made. TREATMENT AND OUTCOME: The dog was treated with prednisone (3.2 mg/kg [1.45 mg/lb], PO, q 24 h), for 3 weeks with limited response. Consequently, azathioprine (2 mg/kg [0.9 mg/lb], PO, q 24 h) was administered. Three weeks later, the dog was evaluated for tachypnea and lethargy. Complete blood count revealed leukopenia, neutropenia, and a left shift. Thoracic radiography revealed a diffuse bronchointerstitial pattern. The dog subsequently went into respiratory arrest and died. On histologic evaluation, amoebic organisms were observed in the lungs, kidneys, and meninges of the brain and spinal cord. A unique Acanthamoeba sp was identified by use of PCR assay. CLINICAL RELEVANCE: This dog developed systemic amoebic infection presumed to be secondary to immunosuppression. The development of secondary infection should be considered in animals undergoing immunosuppression for immune-mediated disease that develop clinical signs unrelated to the primary disease. Although uncommon, amoebic infection may develop in immunosuppressed animals. Use of a PCR assay for identification of Acanthamoeba spp may provide an antemortem diagnosis.     

Severe leukopenia and liver necrosis in young African grey parrots (Psittacus erithacus erithacus) infected with psittacine circovirus

This paper describes the signs, clinical pathology, and postmortem findings in 14 young African grey parrots (Psittacus erithacus erithacus) that were naturally infected with psittacine beak and feather disease (PBFD) virus (psittacine circovirus). All but two of the parrots had severe leukopenia at clinical presentation. Two other parrots also had severe anemia. All birds died within 3 wk after presentation. Postmortem examination documented liver necrosis in 11 of 14 birds and secondary bacterial or fungal infections in 9 of 14 birds. Tests for Chlamydia psittaci, polyomavirus, and Salmonella sp. were negative. PBFD viral infection could be demonstrated in all birds by polymerase chain reaction. Supporting evidence of PBFD viral infection was gathered by histologic examination of the bursa of Fabricius, electron microscopy, and DNA in situ hybridization. Electron microscopic examination of both the bursa of Fabricius and liver revealed virus particles resembling circovirus. DNA in situ hybridization of six liver tissue samples confirmed the presence of PBFD virus and excluded the presence of avian polyomavirus. Our findings suggest that a specific presentation of peracute PBFD viral infection, characterized by severe leukopenia, anemia, or pancytopenia and liver necrosis in the absence of feather and beak abnormalities, may occur in young African grey parrots.     

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.