Idiopathic immune-mediated hemolytic anemia: treatment outcome and prognostic factors in 149 dogs

BACKGROUND: Canine idiopathic immune-mediated hemolytic anemia (IMHA) is associated with a high mortality, especially in the 1st 2 weeks after diagnosis despite treatment. OBJECTIVES: To determine treatment outcome and identify prognostic variables in order to define areas of future research. ANIMALS: One hundred forty-nine dogs with hematocrit <30% and either a positive Coombs' test or spherocytosis and with no evidence of disease that can trigger IMHA were included. METHODS: Retrospective cohort study. All dogs were treated with prednisolone and azathioprine according to a standard protocol. Survival analysis was performed by the Kaplan-Meier method. Variables recorded at the time of diagnosis were tested as possible prognostic variables in a univariate and multivariate Cox proportional hazard model. RESULTS: The main predictors for mortality in dogs with idiopathic IMHA are the presence of increased plasma urea concentration, bands, thrombocytopenia, and petechiae at the time of diagnosis. The estimated Kaplan-Meier half-year survival was 72.6% (95% confidence interval [CI]: 64.9-81.3%). Mortality occurred mostly within the 1st 2 weeks. Cox proportional hazards analysis indicated that increased plasma urea concentration, icterus, and petechiae were the major independent predictors of mortality in the 1st 2 weeks. In most dogs that survived IMHA, a 3-month protocol of azathioprine with prednisolone maintained clinical remission. The estimated half-year survival for dogs that survived the 1st 2 weeks was 92.5% (95% CI: 86-99.3%). CONCLUSIONS AND CLINICAL IMPORTANCE: If the dogs survived IMHA, a 3-month protocol of prednisolone and azathioprine was effective with regard to survival and clinical outcome. Future research should be directed at identifying whether thrombotic tendency in dogs with IMHA is the main contributor to the development of increased plasma urea concentration, icterus, thrombocytopenia, and petechiae.     

Use of serum concentrations of interleukin-18 and monocyte chemoattractant protein-1 as prognostic indicators in primary immune-mediated hemolytic anemia in dogs

Background: The cytokine response in immune‐mediated hemolytic anemia (IMHA) is poorly characterized and correlation with outcome is unknown. Hypothesis/Objectives: To determine if cytokine activity is correlated with outcome in dogs with IMHA. Animals: Twenty dogs with primary IMHA and 6 control dogs. Methods: Prospective study on dogs with IMHA with blood sampling at admission. Serum activity of interleukin‐2 (IL‐2), IL‐4, IL‐6, IL‐7, IL‐8, IL‐10, IL‐15, IL‐18, monocyte chemoattractant protein‐1 (MCP‐1), granulocyte‐macrophage colony stimulating factor (GM‐CSF), interferon‐inducible protein‐10, interferon‐gamma, and keratinocyte chemoattractant (KC) was assessed. Results: Thirty‐day case fatality rate was 25% (5/20 dogs). Increased concentrations (median [range]) of IL‐2 (45.5 ng/L [0;830] versus 0 ng/L [0;46.8]), IL‐10 (8.2 ng/L [0;60.6] versus 0 ng/L [0;88.2]), KC (1.7 μg/L [0.3;4.7] versus 0.5 μg/L [0.2;1.1]), and MCP‐1 (162 ng/L [97.6;438] versus 124 ng/L [90.2;168]) were observed in dogs with IMHA compared with controls. The cytokine profile was indicative of a mixture of pro‐ and anti‐inflammatory cytokines of various cellular origins. Cytokines/chemokines strongly associated with macrophage/monocyte activation and recruitment were significantly increased in nonsurvivors compared with survivors; IL‐15 (179 ng/L [48.0;570] versus 21.3 ng/L [0;193]), IL‐18 (199 ng/L [58.7;915] versus 37.4 ng/L [0;128]), GM‐CSF (134 ng/L [70.0;863] versus 57.6 ng/L [0;164]), and MCP‐1 (219 ng/L [135;438] versus 159 ng/L [97.6;274]), respectively. Logistic regression suggested increased IL‐18 and MCP‐1 concentrations were independently associated with mortality in this population (P<.05, Wald's type 3). Conclusions and Clinical Importance: A mixed cytokine response is present in dogs with IMHA and mediators of macrophage activation and recruitment might serve as prognostic indicators.     

Vaccine-Associated Immune-Mediated Hemolytic Anemia in the Dog

Vaccination has been incriminated as a trigger of immune-mediated hemolytic anemia (IMHA) in dogs and in people, but evidence to support this association is lacking. In a controlled retrospective study, idiopathic IMHA was identified in 58 dogs over a 27–month period. When compared with a randomly selected control group of 70 dogs (presented for reasons other than IMHA) over the same period, the distribution of cases versus time since vaccination was different (P < .05). Fifteen of the dogs (26%) had been vaccinated within 1 month (mean, 13 days; median, 14 days; range, 1 to 27 days) of developing IMHA (P < .0001), whereas in the control group no marked increase in frequency of presentation was seen in the first month after vaccination. The dogs with IMHA were divided into 2 groups based on time since vaccination: the vaccine IMHA group included dogs vaccinated within 1 month of developing IMHA; the nonvaccine IMHA group included dogs that developed IMHA more than 1 month after vaccination. The recently vaccinated dogs with IMHA (vaccine IMHA group) had significantly lower platelet counts (P < .05) and a trend towards increased prevalence of intravascular hemolysis and autoagglutination when compared with the nonvaccine IMHA group. Similar mortality rates were seen in the vaccine IMHA group (60%) and the nonvaccine IMHA group (44%), with the majority of fatalities (>75%) occurring in the first 3 weeks after presentation. Persistent autoagglutination was a negative prognostic indicator for survival in both groups (P < .05). Presence of icterus and hyperbilirubinemia were negative prognostic indicators for survival in the nonvaccine IMHA group (P < .0001 and P < .01, respectively) but not in the vaccine IMHA group. In the recently vaccinated dogs, combination vaccines from various manufacturers against canine distemper, adenovirus type 2, leptospirosis, parainfluenza, and parvovirus (DHLPP) were involved in each case. Vaccines against rabies virus, Bordetella spp, coronavirus, and Lyme Borrelia were administered concomitantly to some dogs. This study provides the first clinical evidence for a temporal relationship of vaccine-associated IMHA in the dog.     

Retrospective Study: The association of blood lactate concentration with outcome in dogs with idiopathic immune‐mediated hemolytic anemia: 173 cases (2003–2006)

Objective – To determine the association of blood lactate with outcome and response to transfusion therapy in dogs with idiopathic immune‐mediated hemolytic anemia (IMHA). Design – Retrospective study. Setting – Urban veterinary small animal emergency hospital. Animals – One hundred and seventy‐three client‐owned dogs with IMHA. Interventions – None. Measurements and Main Results – Serial blood lactate concentration, therapeutic interventions, and outcome were recorded. Nonsurvivors were defined as those that died or were euthanized. One hundred and thirty‐three dogs (77%) survived, 35 (20%) were euthanized, and 5 (3%) died. One hundred forty‐five dogs (84%; 145/173) had a lactate concentration above the laboratory reference interval [0.46–2.31 mmol/L] on presentation. Blood lactate at presentation was higher in the nonsurvivors (median 4.8 mmol/L; 0.5–13.6) compared with survivors (median 2.9 mmol/L; 0.3–13.2) (P<0.01). All dogs presenting with hyperlactatemia that normalized (<2.0 mmol/L) within 6 hours of admission survived, whereas, 71% of dogs that had a persistent hyperlactatemia at 6 hours survived (P=0.034). Lactate was positively correlated with age, BUN, and alkaline phosphatase, and inversely correlated with PCV. Receiver operating curve analysis for lactate concentration at admission as a test for outcome had an area under the curve of 0.69 with an optimal lactate cutoff concentration of 4.4 mmol/L correctly predicting outcome 73% of the time (sensitivity 60%, specificity 77%). Conclusions – Lactate concentration at presentation was significantly higher in nonsurvivors than survivors. Lactate was significantly correlated with previously reported outcome variables but lactate concentration at admission, as a predictor for outcome was less than optimal. However, serial lactate concentration measurements may be more predictive as patients with persistent hyperlactatemia 6 hours after admission were less likely to survive. Prospective studies evaluating serial lactate concentration while controlling for other variables may provide further insight into lactate measurement as a prognostic indicator in animals with IMHA.     

Neutrophil Extracellular Traps in Plasma from Dogs with Immune‐mediated Hemolytic Anemia

Background

Neutrophil extracellular traps (NETs) are part of the innate immune response and are essential in local pathogen control, but are associated with pathological inflammation, organ damage, autoimmunity, and thrombosis. Immune‐mediated hemolytic anemia (IMHA) is a pro‐inflammatory, prothrombotic disease associated with high mortality.

Hypothesis/Objectives

Neutrophil extracellular traps (NETs) are a feature of the inflammatory process in dogs with IMHA. The objective of the study was to evaluate plasma from dogs with IMHA for the presence of 2 indirect markers and 1 direct marker of NETs.

Animals

Healthy client‐owned dogs (56) and hospitalized dogs with IMHA (n = 35).

Methods

Prospective study. Plasma samples for all dogs were evaluated for cell‐free DNA using a fluorescence assay, histone‐DNA (hisDNA) complex using an ELISA, and citrullinated histone H3 (specific for NETosis) using Western blot. Reference intervals were generated using plasma from healthy dogs.

Results

In dogs with IMHA, cell‐free DNA concentration was above the reference interval in 17% of samples with a median (range) of 1.0 μg/mL (0.1–17.3), and hisDNA concentration was above the reference interval in 94% of samples with a median (range) of 30.7 × pooled normal plasma (PNP; 0.6–372.1). Western blot for citrullinated histone H3 identified detectable bands in 84% samples from dogs with IMHA.

Conclusions and Clinical Importance

The assay for cell‐free DNA detected evidence of NETs in fewer dogs than did the other approaches. Excessive NETs appears to be a feature of IMHA in dogs and contributions to the prothrombotic state deserve further study.     

Evaluation of prognostic factors, survival rates, and treatment protocols for immune-mediated hemolytic anemia in dogs: 151 cases (1993-2002)

OBJECTIVE: To evaluate prognostic factors, survival, and treatment protocols for immune-mediated hemolytic anemia (IMHA) in dogs. DESIGN: Retrospective study. ANIMALS: 151 dogs with IMHA not associated with underlying infectious or neoplastic disease. PROCEDURE: lnformation recorded from review of medical records included signalment at the time of initial evaluation; vaccination history; 30-, 60-, and 365-day follow-up outcomes; laboratory data; results of imaging studies; and necropsy findings. Dogs were grouped according to the presence of spherocytes, autoagglutination, a regenerative erythrocyte response, and treatments received (azathioprine, azathioprine plus ultralow-dose aspirin, azathioprine plus mixed-molecular-weight heparin [mHEP], or azathioprine plus ultralow-dose aspirin plus mHEP) for comparisons. All dogs received glucocorticoids. RESULTS: Cocker Spaniels, Miniature Schnauzers, neutered dogs, and female dogs were overrepresented. Alterations in certain clinicopathologic variables were associated with increased mortality rate. Rates of survival following treatment with azathioprine, azathioprine plus ultralow-dose aspirin, azathioprine plus mHEP, and azathioprine plus ultralow-dose aspirin plus mHEP were 74%, 88%, 23%, and 70%, respectively, at hospital discharge; 57%, 82%, 17%, and 67%, respectively, at 30 days; and 45%, 69%, 17%, and 64%, respectively, at 1 year. In comparison, mean survival rates at discharge and at 30 days and 1 year after evaluation collated from 7 published reviews of canine IMHA were 57%, 58%, and 34%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment with a combination of glucocorticoids, azathioprine, and ultralow-dose aspirin significantly improved short- and long-term survival in dogs with IMHA.     

Treatment of Immune‐Mediated Hemolytic Anemia with Individually Adjusted Heparin Dosing in Dogs

Background: A major cause of death in dogs with immune‐mediated hemolytic anemia (IMHA) is thromboembolism. Previous studies suggest unfractionated heparin (UH) is not effective in preventing thromboembolism in IMHA; however, subtherapeutic dosing could explain the seeming lack of efficacy. Hypothesis: Providing therapeutic plasma concentration of UH by individually adjusting doses based on antifactor Xa activity would improve survival in IMHA. Animals: Fifteen dogs with primary IMHA. Methods: Randomized, prospective, controlled clinical trial. Dogs received standardized therapy for IMHA and either constant dose (CD) (150 U/kg SC) (n = 7) or individually adjusted dose (IAD) (n = 8) UH, monitored via an anti‐Xa chromogenic assay, adjusted according to a nomogram. UH was administered every 6 hours until day 7, and every 8 hours thereafter. UH dose was adjusted daily in IAD dogs until day 7, weekly until day 28, then tapered over 1 week. Dogs were monitored for 180 days. Results: At day 180, 7 dogs in the IAD group and 1 in the CD group were alive (P= .01). Median survival time for the IAD group was >180 days, and 68 days for the CD group. Thromboembolic events occurred in 5 dogs in the CD group and 2 dogs in the IAD group. Doses of UH between 150 and 566 U/kg achieved therapeutic anti‐Xa activity (0.35–0.7 U/mL). Conclusions and Clinical Importance: This study suggests that IAD UH therapy using anti‐Xa monitoring reduced case fatality rate in dogs with IMHA when compared with dogs receiving fixed low dose UH therapy.     

Immune‐mediated hemolytic anemia and severe thrombocytopenia in dogs: 12 cases (2001–2008)

Objective – To identify and characterize the syndrome of immune‐mediated hemolytic anemia (IMHA) with concurrent severe thrombocytopenia (≤15.0 × 10⁹ platelets/L; [15.0 × 10³ platelets/μL]), and to evaluate prognostic factors, clinicopathologic findings, complications, treatment, outcome, and survival of dogs with this hematologic disorder. Design – Retrospective, observational study. Setting – Veterinary teaching hospital. Animals – Twelve client‐owned dogs with IMHA and severe thrombocytopenia (≤15.0 × 10⁹ platelets/L; [15.0 × 10³ platelets/μL]), without evidence of overt disseminated intravascular coagulation. Interventions – The following data were recorded and analyzed from the electronic medical record: signalment, history, concurrent diseases, clinical signs at presentation, clinicopathologic data, diagnostic testing, radiographic findings, treatment modalities, length of hospitalization, complications, and clinical outcome. All dogs were treated with immunosuppressive doses of corticosteroids. Measurements and Main Results – Twelve dogs were identified with the diagnosis of IMHA and severe thrombocytopenia; of these, 9 (75%) survived, 3 (25%) were euthanized, and none died. Dogs that survived were significantly younger than nonsurvivors (P=0.03). There were no specific clinical signs or therapies associated with survival. Conclusions – Dogs in this study had a mortality rate similar to reported rates for dogs with either disease alone. Overall, younger dogs were more likely to survive. No association between different treatment modalities and overall survival was identified.     

Evaluation of Risk Factors for Morbidity and Mortality after Pylorectomy and Gastroduodenostomy in Dogs

Objectives— To (1) identify and describe the type and frequency of postoperative complications after pylorectomy and gastroduodenostomy in dogs and (2) identify preoperative and intraoperative risk factors, including the presence of neoplasia, prognostic for patient mortality after surgery. Study Design— Case series. Animals— Dogs (n=24) treated by pylorectomy and gastroduodenostomy. Methods— Medical records (2000–2007) for 2 teaching hospitals of dogs treated that had pylorectomy and gastroduodenostomy were reviewed. Pre‐, intra‐, and postoperative data were obtained from the medical record. Results— Of the 24 dogs, 75% survived 14 days, but 10 (41%) died by 3 months. Overall median survival time (MST) was 578 days. On log‐rank univariate analysis, preoperative weight loss (P=.001) and malignant neoplasia (P=.01) were associated with decreased survival time. Dogs with malignant neoplasia had a MST of 33 days. Common postoperative morbidity included hypoalbuminemia (62.5%) and anemia (58.3%). Conclusions— Pylorectomy with gastroduodenostomy has a good short‐term outcome but long‐term survival time is poor in dogs with malignant neoplasia. Clinical Relevance— Overall, most dogs treated with pylorectomy and gastroduodenostomy survived the postoperative period; however, preoperative weight loss and malignant neoplasia were associated with decreased survival time. Because dogs with malignant neoplasia have markedly shortened survival times, pertinent preoperative, diagnostics steps should be exhausted to identify underlying neoplasia.     

Evaluation of immunosuppressive regimens for immune-mediated haemolytic anaemia: a retrospective study of 42 dogs

Objectives : Immune‐mediated haemolytic anaemia (IMHA) is a severe disease for which evidence is lacking to make informed choices regarding immunosuppressive regimen. The aims of the current study were to determine the effect of different treatment regimens on outcome in affected animals and to identify parameters that may be used as prognostic factors for the disease. Methods : The records of dogs presenting to a veterinary hospital in the period 2002 to 2010 for treatment of IMHA were examined and follow‐up data were obtained. Statistical tests were performed to establish whether treatment regimen affected outcome and to identify prognostic factors for outcome. Results : Treatment regimen had a significant effect on the outcome (measured as survival of hospitalisation) but there were insufficient subjects to determine the cause of the difference. Serum bilirubin and urea concentrations were found to be significant negative prognostic factors for the outcome of IMHA cases and the concentrations of these parameters were significantly different between animals that survived or died while hospitalised. Clinical Significance : This study presents the first report of a significant difference in outcome comparing animals treated with immunosuppressive drugs which are in widespread clinical usage. Although possible confounding factors should be considered, these findings could have major consequences for the treatment of IMHA.     

Canine idiopathic immune-mediated haemolytic anaemia: a review with recommendations for future research

Idiopathic immune-mediated haemolytic anaemia (IMHA) is one of the most common immune-mediated diseases of dogs. The aim of this article is to review current knowledge of canine IMHA, its etiology, clinical presentation, diagnosis, complications, and treatment, in an attempt to establish why its outcome is still so poor. Clinical signs of anaemia develop within 3 days and dogs present with a median haematocrit of 13%, leucocytosis, a left shift, and reticulocytosis. Coagulation test results support the presence of disseminated intravascular coagulation. About 50% of dogs die in the first 2 weeks after presentation, and analysis of risk factors suggests that mortality is associated with hypercoagulability, inflammatory response, and liver and kidney failure. A positive direct agglutination test, spherocytosis, and true autoagglutination are widely accepted tests to demonstrate anti-erythrocyte antibodies, but are not yet standardized. To date, there is no evidence to support the efficacy of immunomodulators in addition to corticosteroids in the treatment of IMHA. Despite numerous investigations, the prognosis of IMHA remains dismal. There is an urgent need to validate and standardize diagnostic tests and criteria, and clinical trials might benefit from stratifying dogs by mortality risk. Analysis of samples from well-defined cases of canine IMHA might provide insight into the aetiology and pathophysiology of IMHA.     

Anti-Endothelial Cell Antibodies in Dogs with Immune-Mediated Hemolytic Anemia and Other Diseases Associated with High Risk of Thromboembolism

Background: Dogs with immune-mediated hemolytic anemia (IMHA) and certain inflammatory diseases are at high risk of developing thromboembolic disease. The presence of anti-endothelial cell autoantibodies (AECA) has been associated with an increased risk of thromboembolism in humans.
Hypothesis: AECA will be detected more often in dogs at risk of thromboembolism than in healthy control animals or dogs with diseases not associated with a higher risk of thromboembolism.
Animals: Ninety-one sick dogs and 22 healthy control dogs.
Methods: Retrospective case-controlled study. Serum was screened for the presence of AECA. Dogs were identified for the study based on the risk of thromboembolism as determined by clinical impression and the underlying disease process. Flow cytometry and normal canine endothelial cells were used to screen serum samples from sick and healthy control dogs for the presence of AECA. In addition, serum from dogs with confirmed thromboemboli was also screened for the presence of AECA by immunohistochemistry.
Results: AECA were detected in 2/91 sick dogs, both with infectious diseases, but were not found in healthy dogs. Anti-endothelial antibodies were not detected in 21 dogs with IMHA and 20 dogs with systemic inflammatory response syndrome, sepsis, or both.
Conclusions: We conclude that AECA are rarely detectable in dogs considered at high risk of thromboembolism. These findings suggest that AECA may not play an important role in the pathogenesis of thromboembolism in dogs with IMHA and other inflammatory diseases.     

Cerebrospinal Fluid Myelin Basic Protein as a Prognostic Biomarker in Dogs with Thoracolumbar Intervertebral Disk Herniation

Background: Release of myelin basic protein (MBP) into the cerebrospinal fluid (CSF) is associated with active demyelination and correlates with outcome in various neurological diseases. Hypothesis/Objectives: To describe associations among CSF MBP concentration, initial neurological dysfunction, and long‐term ambulatory outcome in dogs with acute thoracolumbar intervertebral disk herniation (IVDH). Animals: Five hundred and seventy‐four dogs with acute thoracolumbar IVDH and 16 clinically normal dogs. Methods: Prospective case series clinical study. Signalment, initial neurological dysfunction as determined by a modified Frankel score (MFS), and ambulatory outcome at >3‐month follow‐up were recorded. Cisternal CSF MBP concentration was determined by an ELISA. Associations were estimated between CSF MBP concentration and various clinical parameters. Results: Dogs with thoracolumbar IVDH that did not ambulate at follow‐up had a higher CSF MBP concentration (median, 3.56 ng/mL; range, 0.59–51.2 ng/mL) compared with control dogs (median, 2.22 ng/mL; range, 0–3.82 ng/mL) (P= .032). A CSF MBP concentration of ≥3 ng/mL had a sensitivity of 78% and specificity of 76% to predict an unsuccessful outcome based on receiver‐operating characteristics curve analysis (area under the curve =0.688, P= .079). Affected dogs with a CSF MBP concentration ≥3 ng/mL had 0.09 times the odds of ambulation at follow‐up compared with affected dogs with CSF MBP concentration <3 ng/mL when adjusted for initial MFS (95% confidence interval 0.01–0.66, P= .018). Conclusions and Clinical Importance: These results would suggest that CSF MBP concentration may be useful as an independent prognostic indicator in dogs with thoracolumbar IVDH.     

Retrospective Evaluation of the Effect of Heart Rate on Survival in Dogs with Atrial Fibrillation

Background

Atrial fibrillation (AF) usually is associated with a rapid ventricular rate. The optimal heart rate (HR) during AF is unknown.

Hypothesis/Objectives

Heart rate affects survival in dogs with chronic AF.

Animals

Forty‐six dogs with AF and 24‐hour ambulatory recordings were evaluated.

Methods

Retrospective study. Holter‐derived HR variables were analyzed as follows: mean HR (meanHR, 24‐hour average), minimum HR (minHR, 1‐minute average), maximum HR (maxHR, 1‐minute average). Survival times were recorded from the time of presumed adequate rate control. The primary endpoint was all‐cause mortality. Cox proportional hazards analysis identified variables independently associated with survival; Kaplan‐Meier survival analysis estimated the median survival time of dogs with meanHR <125 bpm versus ≥125 bpm.

Results

All 46 dogs had structural heart disease; 31 of 46 had congestive heart failure (CHF), 44 of 46 received antiarrhythmic drugs. Of 15 dogs with cardiac death, 14 had CHF. Median time to all‐cause death was 524 days (Interquartile range (IQR), 76–1,037 days). MeanHR was 125 bpm (range, 62–203 bpm), minHR was 82 bpm (range, 37–163 bpm), maxHR was 217 bpm (range, 126–307 bpm). These were significantly correlated with all‐cause and cardiac‐related mortality. For every 10 bpm increase in meanHR, the risk of all‐cause mortality increased by 35% (hazard ratio, 1.35; 95% CI, 1.17–1.55; P < 0.001). Median survival time of dogs with meanHR<125 bpm (n = 23) was significantly longer (1,037 days; range, 524‐open) than meanHR ≥125 bpm (n = 23; 105 days; range, 67–267 days; P = 0.0012). Mean HR was independently associated with all‐cause and cardiovascular mortality (P < 0.003).

Conclusions and Clinical Importance

Holter‐derived meanHR affects survival in dogs with AF. Dogs with meanHR <125 bpm lived longer than those with meanHR ≥ 125 bpm.     

Detection of activated platelets in dogs with primary immune-mediated hemolytic anemia

Thromboembolism is a major cause of morbidity and mortality in dogs with immune-mediated hemolytic anemia (IMHA). To the authors' knowledge, the role of platelets in thromboembolic events associated with IMHA has not been extensively investigated. In the study reported here, we evaluated cell membrane expression of P-selectin with flow cytometry to determine whether platelets circulate in an activated state in association with primary IMHA. Median P-selectin expression for 20 dogs with primary IMHA was 8.1-fold greater, compared with values for 20 healthy dogs. Fifteen of 20 dogs (75%) with IMHA had P-selectin median fluorescence intensity (MFI) values that exceeded the reference interval for healthy dogs. Additionally, P-selectin MFI after activation of platelets with phorbol myristate acetate was 2.1-fold greater for dogs with IMHA than for healthy control dogs. Despite treatment of all dogs with immunosuppressive therapy and 18 dogs with subcutaneously administered low-dose unfractionated heparin, 7 dogs developed clinical signs consistent with thromboembolism. These data provide support for the hypothesis that platelets circulate in an activated state in many dogs with IMHA.     

Analysis of risk factors associated with recurrence of canine babesiosis caused by Babesia gibsoni

Canine babesiosis due to Babesia gibsoni (B. gibsoni) displays severe clinical manifestations. Recurrence of babesiosis after anti-babesial treatment is observable in over 10 % of the patients. The present study ascertains the risk factors and cumulative incidence of recurrence of canine babesiosis. For a sample of 145 dogs diagnosed with acute babesiosis, the following parameters were assessed over a period of 16 weeks: haematological parameters, status of anaemia, platelet count, total WBC count, haemoglobin concentration and RBC count, concurrent haemoparasitism, and secondary immune mediated haemolytic anaemia (IMHA). Patient demographics such as age, breed, sex were also recorded. The potential risk factors were statistically evaluated by the cumulative incidence function and the Kaplan-Meier method. The recurrent infections were observed in 11.8 % of the study sample. The following factors were found to associate with increased risk of recurrence: Rottweiler breed (CIR 21.8 % ± 6.9 %; p < 0.05), secondary IMHA (CIR 28.7 % ± 11.3 %; p < 0.05), RBC counts < 2 × 10⁶/μl on the day of diagnosis (CIR 16 % ± 4.6 %; p < 0.05), and persistent anaemia over 20 days post treatment (CIR 29.14 ± 7.9 %; p < 0.001). Dogs with concurrent haemoparasitic infections were predicted to have a fatal outcome in the survival analysis (disease related mortalities 25 % ± 13 %; p < 0.001). According to the findings, veterinarians need to pay attention to Rottweiler breed, dogs with secondary IMHA, concurrent haemoparasitism, low RBC counts on diagnosis and those with persistent anaemia to reduce the risk of relapse.     

Peripheral Nucleated Red Blood Cells as a Prognostic Indicator in Heatstroke in Dogs

Background: Heatstroke in dogs is often fatal and is associated with a high prevalence of secondary complications. Peripheral nucleated red blood cells (NRBC) occur in dogs with heatstroke, but their association with complications and the outcome is unclear.
Hypothesis: Peripheral NRBC are common in dogs with heatstroke and have prognostic significance.
Animals: Forty client-owned dogs with naturally occurring heatstroke.
Methods: Prospective, observational study. Dogs were followed from presentation to discharge or death. Serum biochemistry and coagulation tests were performed at presentation. CBC and evaluation of peripheral blood smears were performed at presentation and every 12 hours. The relative and the absolute NRBC numbers were calculated.
Results: Presence of NRBC was observed in 36/40 (90%) of the dogs at presentation. Median relative and absolute NRBC were 24 cells/100 leukocytes (range 0–124) and 1.48 × 10³/μL (range 0.0–19.6 × 10³/μL), respectively. Both were significantly higher in nonsurvivors (22) versus survivors (18) and in dogs with secondary renal failure and DIC versus those without these complications. Receiver operator curve analysis of relative NRBC at presentation as a predictor of death had an area under curve of 0.92. A cut-off point of 18 NRBC/100 leukocytes corresponded to a sensitivity and specificity of 91 and 88% for death.
Conclusions and Clinical Importance: Relative and absolute numbers of peripheral NRBC are clinically useful, correlate with the secondary complications, and are sensitive and specific markers of death in dogs with heatstroke, although they should never be used as a sole prognostic indicator nor should they replace clinical assessment.     

Serial assessment of the coagulation status of dogs with immune-mediated haemolytic anaemia using thromboelastography

This study investigated the coagulation status of dogs with immune-mediated haemolytic anaemia (IMHA) over time. Thirty animals with primary IMHA were blood sampled on three occasions over a 5 day period and assays performed included prothrombin time, activated partial thromboplastin time, D-dimer and fibrinogen concentration, antithrombin activity and recalcified unactivated thromboelastography (TEG). Based on TEG, dogs with IMHA were significantly hypercoagulable vs. controls (P<0.001) and over the 5 day period, 3/4 of the TEG parameters reflected increased clotting kinetics (P ≤ 0.02). The 30 day survival of these patients was 80% and, at hospital admission, the TEG maximum amplitude (MA) was significantly higher in survivors than non-survivors (P=0.015). Each unit increase in MA was associated with an increased odds of 30 day survival of 1.13 (95%; CI 1.02-1.25). Based on TEG, most dogs with IMHA were hypercoagulable on admission and their clotting kinetics increased with time. Relative hypocoagulability identified by TEG at initial assessment was found to be a negative prognostic indicator.     

Immune-mediated haemolytic anaemia: possible association with Ancylostoma caninum infection in three dogs

Immune-mediated haemolytic anaemia (IMHA) may be primary or secondary. In primary IMHA, no underlying cause can be found, whereas secondary IMHA is triggered by an underlying cause, such as neoplasia, infectious diseases, or drugs. This paper describes 3 dogs with typical signs of IMHA that was possibly associated with the intestinal parasite Ancylostoma caninum. As intestinal helminths can be difficult to diagnose on faecal examination, it would be pertinent to perform multiple faecal examinations on any animal that has IMHA with no apparent underlying cause, as part of the therapy.     

Antimicrobial Resistance Impacts Clinical Outcome of Granulomatous Colitis in Boxer Dogs

Background: Escherichia coli have recently been identified within the colonic mucosa of Boxer dogs with granulomatous colitis (GC). Eradication of invasive E. coli is associated with clinical and histological remission. Objectives: To determine antimicrobial susceptibility profiles of E. coli strains from GC and healthy dogs, and the association of antimicrobial resistance with clinical outcome. Animals: Fourteen Boxer dogs with GC and 17 healthy pet dogs. Methods: Prospective study: E. coli was cultured from GC biopsies and rectal mucosal swabs of healthy dogs. Individual strains were selected by phylogroup and overall genotype, determined by triplex‐ and random amplified polymorphic DNA‐polymerase chain reaction respectively. Antimicrobial susceptibility was determined by broth microdilution minimal inhibitory concentration. Results: Culture yielded 23 E. coli strains from GC (1–3/dog, median 2) and 34 strains from healthy (1–3/dog, median 2). E. coli phylogroups were similar (P= .18) in GC (5A, 7B1, 5B2, 6D) and healthy (2A, 10B1, 15B2, 7D). Resistance to ampicillin, amoxicillin‐clavulanate, cefoxitin, tetracycline, trimethoprim‐sulfa (TMS), ciprofloxacin, and chloramphenicol was greater (P < .05) in GC (21–64%) than healthy (0–24%). Enrofloxacin resistant E. coli were isolated from 6/14 GC versus 0/17 healthy (P= .004). Of the enrofloxacin resistant cases, 4/6 were also resistant to macrophage‐penetrating antimicrobials such as chloramphenicol, rifampicin, and TMS. Enrofloxacin treatment before definitive diagnosis was associated with antimicrobial resistance (P < .01) and poor clinical outcome (P < .01). Conclusions and Clinical Importance: Antimicrobial resistance is common among GC‐associated E. coli and impacts clinical response. Antimicrobial therapy should be guided by mucosal culture and antimicrobial susceptibility testing rather than empirical wisdom.