Hemolytic Anemia In Dogs Research Articles

Methylprednisolone alone or combined with cyclosporine or mycophenolate mofetil for the treatment of immune-mediated hemolytic anemia in dogs, a prospective study.

Benefit of adding a second-line immunosuppressive drug to glucocorticoids for the treatment of non-associative immune-mediated hemolytic anemia (naIMHA) in dogs has not been defined prospectively. Evaluate the effectiveness of different immunosuppressive protocols in naIMHA dogs. Forty-three client-owned dogs. Open label, randomized, clinical trial. Dogs were treated with methylprednisolone (M-group), methylprednisolone plus cyclosporine (MC-group) or methylprednisolone plus mycophenolate mofetil (MM-group). Dogs were defined as responders by disappearance of signs of immune-mediated destruction and hematocrit stabilization. Frequency of responders was compared between M-group and combined protocols (MC and MM-group evaluated together), and among the 3 different therapeutic groups at 14 (T14), 30 (T30), 60 (T60) days after admission. Frequency of complications, length of hospitalization and relapse were also compared. Death rate was evaluated at discharge, T60 and 365 (T365) days. Proportion of responders was not significantly different between M-group and combined protocols (MC and MM-groups), nor among the 3 therapeutic groups at T14, T30, and T60 (P > .17). Frequency of relapse, complications, and length of hospitalization were not significantly different between M-group and dogs treated with combined protocols, nor among the 3 treatment groups (P > .22). Death was significantly more common only for MM-group compared with MC-group at T60 (+42.8%; 95% CI: 11.5-67.4; P = .009), and at T365 (+50%; 95% CI: 17.5-73.2; P = .003). Combined immunosuppressive therapy did not improve hematological response in naIMHA.

The Indirect Antiglobulin (Coombs') Test Is Specific but Less Sensitive Than the Direct Antiglobulin Test for Detecting Anti-Erythrocytic Autoantibodies and Thereby Immune-Mediated Hemolytic Anemia in Dogs.

The immunodiagnostic assessment of dogs suspected of having immune-mediated hemolytic anemia (IMHA) is based on persistent autoagglutination of erythrocytes (after three saline washes), marked spherocytosis, and a positive direct antiglobulin (Coombs') test (DAT). However, the value of using the indirect antiglobulin test (IAT) for the detection of anti-erythrocytic autoantibodies in serum from dogs suspected of having IMHA is unclear. To evaluate the IAT, leftover serum samples from a large cohort of 94 dogs suspected of having IMHA and for which DAT results were known were incubated with DAT- erythrocytes, and five IAT techniques were performed (in part with different reagents and temperatures): microtiter plate (MICRO), microcapillary, laboratory gel column, gel minitube kit (GEL KIT), and immunochromatographic strip kit. Two IAT techniques (MICRO at 37 °C and GEL KIT with rabbit anti-dog polyvalent reagent) detected autoantibodies against erythrocytes in serum from 53% and 57% of DAT+ dogs, respectively, while other IATs performed less well. Moreover, while the analytic specificity of the IAT methods compared to the DAT ranged from 96-100%, the sensitivity range was only 9-57%. Thus, we still recommend DAT for diagnosis and monitoring of IMHA in dogs but conclude that a positive IAT result may aid diagnostically when serum is available, but fresh red blood cells are not.

EFFECTS OF IMIDOPYRAN AND PREDNISONE IN THE TREATMENT OF BABESIOSIOSIS-ASSOCIATED ANEMIA IN DOGS

Aim. To investigate the development and treatment with imidopyran and prednisolone of hemolytic anemia in dogs caused by the protozoan parasite Babesia canis. Methods. 17 domestic dogs weighing 5-10 kg aged 2-5 years were used for the study. The parameters of the general blood analysis were determined using the MicroCC-20 Plus automated hematology analyzer (HTI, USA). Microscopy with a Leica DM4 electric microscope (Germany) was carried out to study the condition of erythrocytes, counting the number of leukocytes and platelets. Results. The main indicator of the development of anemia in animals is the number of erythrocytes, hemoglobin, and hematocrit. Development of babesiosis lead to the hemolytic anemia investigated in dogs before treatment: the number of erythrocytes lower than normal by 20-30%, the level of hemoglobin 40-55%, the average concentration of hemoglobin in erythrocytes 10- 18%, hematocrit 20-30% and the number of platelets 40-50%. Conclusions. The results of the study showed that treatment with imidopyran and prednisolone is effective in cases of babesiosis for dogs caused by the protozoan parasite Babesia canis. Moreover, such treatment decreases the risks of the anemic state development for these animals.

Retrospective analysis of immunosuppressive and anti-thrombotic protocols in nonassociative immune mediated hemolytic anemia in dogs.

Evidence supporting the effectiveness of therapeutic protocols for nonassociative immune-mediated hemolytic anemia (na-IMHA) is weak. Investigate the efficacy of various drugs in na-IMHA. Two hundred forty-two dogs. Multi-institutional retrospective study (2015-2020). Immunosuppressive effectiveness was determined by time to packed cell volume (PCV) stabilization and duration of hospitalization through analysis by mixed model linear regression. Occurrence of disease relapse, death, and antithrombotic effectiveness, were analyzed using mixed model logistic regression. Use of corticosteroids vs a multi-agent protocol had no effect on time to PCV stabilization (P=.55), duration of hospitalization (P=.13), or case fatality (P=.06). A higher rate of relapse (P=.04; odds ratio: 3.97; 95% confidence interval [CI]: 1.06-14.8) was detected in dogs receiving corticosteroids (11.3%) during follow-up (median: 28.5 days, range: 0-1631 days) compared to multiple agents (3.1%) during follow up (median: 47.0 days, range: 0-1992 days). When comparing drug protocols, there was no effect on time to PCV stabilization (P=.31), relapse (P=.44), or case fatality (P=.08). Duration of hospitalization was longer, by 1.8 days (95% CI: 0.39-3.28 days), for the corticosteroid with mycophenolate mofetil group (P=.01) compared to corticosteroids alone. Use of clopidogrel vs multiple agents had no effect on development of thromboses (P ≥ .36). Addition of a second immunosuppressive agent did not alter immediate outcome measures but might be associated with a reduction in relapse. Use of multiple antithrombotic agents did not reduce incidence of thrombosis.

Carboxyhemoglobin as a diagnostic and prognostic biomarker of hemolytic anemias in dogs.

Endogenous production of carbon monoxide during hemoglobin metabolism leads to the formation of carboxyhemoglobin. Carboxyhemoglobin concentration is abnormally high in humans with hemolytic anemia (HA). Measurement of carboxyhemoglobin concentration can discriminate HA from other forms of anemia. Twenty-seven dogs with HA (immune-mediated HA, n=22; microangiopathic HA, n=5), 27 dogs with non-HA (kidney disease, n=14; immune-mediated thrombocytopenia, [n=6]; miscellaneous, n=7) and 24 nonanemic control dogs. Prospective cohort study. Carboxyhemoglobin quantification, a CBC and biochemistry profile were performed upon admission, and survival to hospital discharge and at 30 days were the measured outcomes. Groups were compared by the Mann-Whitney and Kruskal-Wallis tests. Receiver-operator characteristic (ROC) analyses were used to examine the predictive utility of carboxyhemoglobin for the diagnosis of HA in anemic dogs. Carboxyhemoglobin (median [interquartile range]) differed between dogs with HA (7.7% [2.5%]) and non-HA (3.6% [1.05]; P < .001) and dogs with HA and nonanemic dogs (3.5% [0.65%]; P < .001). No difference was detected between nonHA and nonanemic dogs. The area under the ROC curve for carboxyhemoglobin as predictor of HA in anemic dogs was 0.997 (95% CI, 0.99-1.00). Three optimal cut-off points were identified, including 5.05%, 4.55% and 4.85%, with corresponding sensitivity/specificity of 92.6%/100%, 100%/92.6% and 96.3%/96.3%, respectively. Neither carboxyhemoglobin nor any of the CBC or chemistry analytes were associated with survival. Carboxyhemoglobin proved an excellent predictor of HA in dogs and might constitute a useful, ancillary tool for diagnosing and monitoring hemolytic anemias.

PetSavers interview

Oliver Garden, the Henry and Corinne R Bower Professor of Medicine and Chair of the Department of Clinical Sciences &amp; Advanced Medicine at the University of Pennsylvania (Penn Vet), has received three PetSavers’ grants – two as the supervisor of undergraduate students and one as the co-applicant of a clinical research project with Dr Barbara Glanemann investigating single versus split-dose regimens of prednisolone in treating immune-mediated haemolytic anaemia in dogs.

O exame de sangue e a anemia hemolítica em cães e gatos – revisão

Precise diagnosis and treatment of hemolytic anemia is a challenge to veterinarians. Toxins, infectious, and autoimmune diseases can cause hemolytic anemia in dogs and cats. Blood smears provide information that can assist in the diagnosis and help to identify the possible etiology of the condition. In many veterinary clinics, the introduction of automated blood cell count has led to abandoning the routine of evaluating blood smears. This article reviews the citologic characteristics of different types of hemolytic anemias in dogs and cats.

Effect of dilution of canine blood samples on the specificity of saline agglutination tests for immune-mediated hemolysis.

BackgroundSaline agglutination tests (SATs) are widely recommended for diagnosis of immune‐mediated hemolytic anemia in dogs, but there are frequent false‐positive results.ObjectivesSpecificity of SATs will improve at higher saline‐to‐blood ratios.AnimalsOne hundred fifty dogs treated at a veterinary referral hospital with hematocrits ≤30%.MethodsProspective diagnostic accuracy study. Immune‐mediated hemolysis (IMH) was considered present if a gel direct antiglobulin test (DAT) was positive and there was clinical evidence of hemolysis (n = 9), absent if another mechanism for anemia was identified and the DAT was negative or there was no hemolysis (n = 138), and if IMH status was unclear, dogs were excluded (n = 3). Saline agglutination tests were prepared at 1 : 1, 4 : 1, 9 : 1, and 49 : 1 saline‐to‐blood ratios, and microscopic agglutination was considered a positive result.ResultsSpecificity for IMH increased from 29% (95% confidence interval 20‐38) at a 1 : 1 dilution to 97% (93‐99) at a 49 : 1 dilution. Sensitivity was 88% (47‐100) at 1 : 1 and 4 : 1 dilutions and 67% (30‐93%) at 9 : 1 and 49 : 1 dilutions. Diagnostic accuracy increased from 33% (24–42) at 1 : 1 dilution to 95% (90‐98) at 49 : 1 dilution.Conclusions and Clinical ImportanceIf performed using a 49 : 1 saline‐to‐blood ratio, SATs achieve high specificity for IMH. Based on a gold standard of positive DAT and evidence of hemolysis, lower saline‐to‐blood ratio results should not be used because false‐positive results are common.

Does vaccination increase the risk of immune-mediated haemolytic anaemia in dogs?

Bottom line You admitted Scout, a four-year-old female neutered English springer spaniel, yesterday as she was jaundiced, anaemic and collapsed. Her haematocrit was 13 per cent, with prominent spherocytosis on...

The utility of screening diagnostic tests in identifying associative immune-mediated haemolytic anaemia in dogs.

The study aimed to (1) define the proportion of dogs with immune-mediated haemolytic anaemia (IMHA) that have associative and non-associative disease and (2) evaluate the utility of screening diagnostic tests in identifying potential triggers of associative IMHA. Medical records of 78 dogs diagnosed with IMHA at a specialist hospital in Sydney from July 2008 to August 2017 were reviewed. The original diagnosis was revised according to published guidelines (Garden et al., 2019) as either diagnostic, supportive or suspicious for IMHA. Associative IMHA was confirmed if immunosuppressive therapy was discontinued within six weeks of effective treatment of a potential trigger. Associative IMHA was considered possible when a potential trigger was identified but its significance could not be confirmed. Associative IMHA was confirmed (3) or suspected (7) in 10 dogs (13%, confidence interval [CI] 7.1%-22%), with 68 cases presumed to be non-associative. Associative IMHA was present in 3/29 (10.3%) of dogs with criteria diagnostic for IMHA, 4/42 (9.5%) of dogs with criteria supportive for IMHA and 3/7 (42.9%) of dogs with criteria suspicious for IMHA. Abdominal ultrasound was performed in 68 dogs and identified possible triggers in five (7.3%, CI 3.2% to 16%). Thoracic radiographs were performed in 70 dogs but did not identify any potential triggers (0%, CI 0% to 5.2%). Urine culture was performed in 22 dogs and was positive in three (14%, CI 4.7% to 33.3%). Routine screening tests, particularly thoracic radiographs, have a low yield in identifying potential triggers of associative IMHA, but are more likely to be useful in dogs fulfilling less stringent diagnostic criteria of IMHA.

Immune-Mediated Hemolytic Anemia Associated with Candidatus Mycoplasma Haematoparvum in a Splenectomized Dog in Italy

Abstract This report describes a case of canine hemotropic mycoplasmasosis by Candidatus Mycoplasma haematoparvum in a dog. A five-year-old splenectomized dog was referred to the Veterinary Teaching Hospital of the University of Sassari with clinical symptoms and laboratory findings compatible with immune-mediated hemolytic anemia. Epicellular bacteria were detected in the erythrocytes by microscopic examination of blood smears. PCR and sequencing were positive for Candidatus Mycoplasma haematoparvum. Treatment with doxycycline, prednisolone and blood transfusion was administered. Several studies have described the molecular prevalence of M. hemocanis and Candidatus M. haematoparvum, however there are few clinical reports, especially those describing Candidatus M. haematoparvum infection in dogs, for which only two cases have been reported. To the best of our knowledge this is the first case report of a symptomatic infection caused by Candidatus Mycoplasma haematoparvum in Italy. Hemoplasmosis should be considered as a potential cause of hemolytic anemia in dogs. Following treatment with doxycycline and prednisolone, the clinical signs improved without resolution of infection. This condition was the same at the three-year follow-up.

Oxidative Stress and Haemolytic Anaemia In Dogs and Cats: A Comparative Approach

Oxidative Stress and Haemolytic Anaemia In Dogs and Cats: A Comparative Approach

Prevalence of Babesia spp. and clinical characteristics of Babesia vulpes infections in North American dogs.

BackgroundBabesiosis is an important cause of thrombocytopenia and hemolytic anemia in dogs. Babesia vulpes, reported in European dogs and North American foxes, rarely has been reported in domestic North American dogs. Newly optimized polymerase chain reaction (PCR) primers facilitate more sensitive amplification of B. vulpes DNA.ObjectivesTo determine the prevalence of Babesia sp. infections in dogs being tested for Babesia infection, and to describe co‐infections and clinicopathologic abnormalities in B. vulpes positive dogs.AnimalsDog blood or tissue samples (n = 9367) submitted to a diagnostic laboratory between June 2015 and June 2018 were tested using an optimized Babesia PCR assay.MethodsComprehensive canine vector‐borne disease diagnostic testing was performed on convenience samples.Results Babesia sp. DNA was amplified from 269/9367 (2.9%) North American dogs. Babesia sp. infections included B. gibsoni monoinfection (157; 1.7%), B. vulpes monoinfection (19; 0.20%), and B. gibsoni and B. vulpes coinfection (29; 0.31%). Forty‐three of the 48 total B. vulpes‐infected dogs were American Pit Bull Terrier‐type breeds, of which 36 historically were involved with dog fights. Coinfections with Mycoplasma, Dirofilaria immitis, or Wolbachia and coexposures to Bartonella, Ehrlichia, and Rickettsia spp. were documented in B. vulpes‐infected dogs. Clinicopathologic data in B. vulpes‐infected dogs both with and without coinfections included anemia, thrombocytopenia, hyperglobulinemia, hypoalbuminemia, and proteinuria.Conclusions and Clinical Importance Babesia vulpes infection in domestic North American dogs is commonly found in conjunction with other coinfections, including B. gibsoni and hemotropic Mycoplasma. Similar to B. gibsoni, dog‐to‐dog transmission of B. vulpes may be a frequent mode of transmission.

ACVIM consensus statement on the treatment of immune-mediated hemolytic anemia in dogs.

Immune‐mediated hemolytic anemia (IMHA) causes severe anemia in dogs and is associated with considerable morbidity and mortality. Treatment with various immunosuppressive and antithrombotic drugs has been described anecdotally and in previous studies, but little consensus exists among veterinarians as to the optimal regimen to employ and maintain after diagnosis of the disease. To address this inconsistency and provide evidence‐based guidelines for treatment of IMHA in dogs, we identified and extracted data from studies published in the veterinary literature. We developed a novel tool for evaluation of evidence quality, using it to assess study design, diagnostic criteria, explanation of treatment regimens, and validity of statistical methods. In combination with our clinical experience and comparable guidelines for humans afflicted with autoimmune hemolytic anemia, we used the conclusions of this process to make a set of clinical recommendations regarding treatment of IMHA in dogs, which we refined subsequently by conducting several iterations of Delphi review. Additionally, we considered emerging treatments for IMHA in dogs and highlighted areas deserving of future research. Comments were solicited from several professional bodies to maximize clinical applicability before the recommendations were submitted for publication. The resulting document is intended to provide clinical guidelines for management of IMHA in dogs. These guidelines should be implemented pragmatically, with consideration of animal, owner, and veterinary factors that may vary among cases.

ACVIM consensus statement on the diagnosis of immune-mediated hemolytic anemia in dogs and cats.

Immune‐mediated hemolytic anemia (IMHA) is an important cause of morbidity and mortality in dogs. IMHA also occurs in cats, although less commonly. IMHA is considered secondary when it can be attributed to an underlying disease, and as primary (idiopathic) if no cause is found. Eliminating diseases that cause IMHA may attenuate or stop immune‐mediated erythrocyte destruction, and adverse consequences of long‐term immunosuppressive treatment can be avoided. Infections, cancer, drugs, vaccines, and inflammatory processes may be underlying causes of IMHA. Evidence for these comorbidities has not been systematically evaluated, rendering evidence‐based decisions difficult. We identified and extracted data from studies published in the veterinary literature and developed a novel tool for evaluation of evidence quality, using it to assess study design, diagnostic criteria for IMHA, comorbidities, and causality. Succinct evidence summary statements were written, along with screening recommendations. Statements were refined by conducting 3 iterations of Delphi review with panel and task force members. Commentary was solicited from several professional bodies to maximize clinical applicability before the recommendations were submitted. The resulting document is intended to provide clinical guidelines for diagnosis of, and underlying disease screening for, IMHA in dogs and cats. These should be implemented with consideration of animal, owner, and geographical factors.

Safety and efficacy of aged garlic extract in dogs: upregulation of the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway and Nrf2-regulated phase II antioxidant enzymes

BackgroundPlants of Allium spp., including garlic (A. sativum) and onions (A. cepa), are known to be oxidatively toxic to canine erythrocytes resulting in Heinz body hemolytic anemia in dogs. In humans, these plants have been used as medicinal agents for multiple diseases since ancient times. Especially, fresh garlic extracted over a prolonged period produces less irritative and odorless aged garlic extract (AGE), containing unique and beneficial organosulfur compounds that can help prevent many kinds of diseases. In this study, the safety and efficacy of long-term oral administration of AGE is evaluated in dogs. The objectives are to confirm the safe dosage for long-term use and beneficial functions of AGE for dogs and to plan and design a canine health supplement or a preventive agent for multiple diseases based on the data of this study.ResultsBeagles were orally administered AGE (45 or 90 mg/kg body weight once a day) or an equivalent amount of water as control for 12 weeks. In AGE-treated groups, at 12 weeks post-administration at a dose of 90 mg/kg, there were no observable changes in the clinical signs, complete blood count, and serum biochemical parameters. Heinz bodies and eccentrocytes, the markers of oxidative damage in erythrocytes, did not appear in blood smear examination. In order to further evaluate the beneficial effects of AGE on health of dogs, the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) gene (NFE2L2) and Nrf2-regulated phase II antioxidant enzyme genes (NQO1, GCLM, HMOX1, and SOD2) were determined in whole blood between pre- and post-AGE administration. The expression of NFE2L2 gene was significantly upregulated in the AGE-treated groups [45 (p < 0.05) and 90 mg/kg (p < 0.01), 8 weeks] as compared to in the control group. Among the Nrf2-regulated enzymes examined, the expressions of NQO1 [45 (p < 0.05) and 90 mg/kg (p < 0.01), 8 weeks] and GCLM [45 (p < 0.05) and 90 mg/kg (p < 0.01), 12 weeks] genes were significantly upregulated.ConclusionThe long-term oral administration of AGE at a dose of 90 mg/kg/day for 12 weeks did not show any adverse effects in dogs. Furthermore, the administration of AGE upregulated the gene expressions of canine Nrf2 and Nrf2-regulated phase II antioxidant enzymes. These results suggest that AGE might safely contribute to the health of dogs provided that the appropriate dosage is used.

Efficacy of the combination of glucocorticoids, mycophenolate-mofetil and human immunoglobulin for the therapy of immune mediated haemolytic anaemia in dogs

Idiopathic immune-mediated haemolytic anaemia (IMHA) is one of the most common immune-mediated diseases in dogs with a high mortality rate. In this retrospective study, we examined the effect of mycophenolate- mofetil (MMF) and human intravenous immunoglobulin (hIVIG) in combination with glucocorticoids on canine IMHA patients. Six dogs were treated with prednisolone and MMF (hIVIG-) and in 15 patients hIVIG was added (hIVIG+). There was no significant difference between the groups regarding age, weight, number of blood transfusions or hematocrit on the day of diagnosis. The hIVIG+ group showed a significantly faster recovery of the hematocrit, but this difference was only short-lived. The survival in the first year was similar in both groups and was with 71.5% somewhat higher than in other published studies. The addition of MMF to prednisolone for the treatment of dogs with acute IMHA was well tolerated and seemed to positively affect the course of the disease. Randomized studies are necessary to confirm this observation. Human immunoglobulin had only minimal clinical advantages and no effect on mortality.

Efficacy of Microtiter Method of Direct Antiglobulin Test in diagnosis of Immune Mediated Haemolytic Anaemia in Dogs

Efficacy of Microtiter Method of Direct Antiglobulin Test in diagnosis of Immune Mediated Haemolytic Anaemia in Dogs

Seasonality of immune-mediated hemolytic anemia in dogs from southern California.

To determine if there is seasonal variation in the incidence of canine idiopathic immune-mediated hemolytic anemia (IMHA) at 2 specialty hospitals in southern California. Retrospective study. To determine seasonality, cases of idiopathic IMHA presenting between June 2006 and June 2012 were identified by searching medical record databases of 2 large specialty hospitals. The presence of anemia with autoagglutination, spherocytosis or a positive Coomb's test and no identifiable underlying cause were required for inclusion. Dogs with a history of recent travel (within 1 year) were excluded. One hundred and twenty-six dogs, 57 from a practice in San Diego, and 69 from a practice in Los Angeles, met the criteria. The pattern of seasonality differed with practice location (P = 0.02). More cases of idiopathic IMHA presented during spring and summer (n = 39) than winter and fall (n = 18) for the San Diego practice (P = 0.008) but not the Los Angeles practice (n = 32 and 37, respectively, P = 0.63). The incidence of idiopathic IMHA in dogs presenting to specialty hospitals located in 2 different climates of southern California was different, suggesting environmental triggers may be involved. Larger, prospective studies are needed to determine whether environmental parameters or undetected infectious disease account for some cases of idiopathic IMHA in dogs.

Systematic Review of Evidence Relating to the Treatment of Immune‐Mediated Hemolytic Anemia in Dogs

Despite being the most prevalent autoimmune disease of dogs, there is considerable variation between individuals and institutions in the treatment regimens that are employed for the management of immune-mediated hemolytic anemia. The aim of this review was to evaluate evidence relating to the treatment of the disease systematically and to use this evidence to draw conclusions that are applicable in wider veterinary practice. Search tools were employed to identify relevant articles and these were assessed according to stated criteria. The overall quality of published evidence was poor, with many articles failing to provide details of the enrollment, treatment, monitoring, and assessment stages of the study process. In view of this, firm conclusions cannot be drawn regarding the treatment of this disease and further research of a higher quality is required.