Immune-mediated hemolytic anemia and pure red cell aplasia in a Jack Russell Terrier during treatment for hypoadrenocorticism.

An 11-year-old neutered male Jack Russell Terrier was presented to Yuki Animal Hospital for regenerative anemia during the treatment of hypoadrenocorticism. A blood smear examination showed spherocytes, polychromatic erythrocytes, and erythrocyte ghosts. The direct agglutination test was positive at 37°C. The dog was then diagnosed with immune-mediated hemolytic anemia (IMHA). Although prednisolone and mycophenolate mofetil were administered, the hematocrit and reticulocyte count decreased, and nonregenerative anemia developed. A bone marrow examination was performed to diagnose the cause of the nonregenerative anemia. Histologic and cytologic bone marrow examination revealed a normocellular to hypercellular medulla with severe erythroid hypoplasia. No proliferation of lymphocytes or lymphoblast-appearing cells was observed. This dog was diagnosed with pure red cell aplasia (PRCA). Despite treatment with immunosuppressive agents, the patient died of thrombosis. Although these associations were unclear, this is the first report of PRCA diagnosis following IMHA and while treating hypoadrenocorticism.

Complete remission of two canine cases with precursor-targeted immune-mediated anemia after combination therapy with prednisolone, cyclosporine, and oclacitinib.

Background: Precursor-targeted immune-mediated anemia (PIMA) has been described in dogs presenting with nonregenerative anemia and evidence of ineffective erythropoiesis. Although it has been suggested that its occurrence may be related to the immune targeting of erythroid precursors, this pathogenesis has not been established. PIMA is mainly treated with glucocorticoids, and in cases where glucocorticoids alone are not effective, immunosuppressants are also used as combination therapy. However, not all cases of PIMA go into remission after these treatments. Case Description: Two dogs with severe nonregenerative anemia diagnosed as PIMA based on the results of clinical pathological examinations, including bone marrow examination, were treated with whole-blood transfusion and immunosuppressive doses of prednisolone, mycophenolate mofetil, and cyclosporine. However, these treatments failed to achieve remission of PIMA. Therefore, concomitant administration of oclacitinib, which is a Janus kinase-1 inhibitor that has been applied recently to the treatment of immune-mediated diseases, was performed; this combined regimen improved the anemia and achieved complete remission of PIMA. Conclusion: Oclacitinib may be an option for the treatment of PIMA in dogs failing to achieve remission with conventional immunosuppressive therapy.

Development of acute pancreatitis after oral administering a praziquantel, pyrantel pamoate, and febantel combination in a dog: A case report.

Oral praziquantel, pyrantel pamoate, and febantel combination (PPFC) is a highly safe anthelmintic treatment commonly administered for the purpose of canine gastrointestinal parasites with mild adverse effects such as anorexia, vomiting, lethargy, or diarrhea. A 12-year-old castrated Chihuahua was brought to our hospital for a periodic health examination. Although his general physical examination showed no abnormalities, blood test results showed increase in the liver enzyme, lipase activity, total bile acid, total cholesterol, and triglyceride concentration. Moreover, the dog had underlying tricuspid regurgitation that was not treated. PPFC was prescribed on the suspicion of gastrointestinal tract parasites. Following the oral administration of PPFC at home, anorexia and lethargy were found, and vomiting and diarrhea were noted after 30 h. The dog was diagnosed with acute pancreatitis based on clinical course of the disease and subsequent pathology results. Although intravenous drip was initiated upon hospitalization, the treatment was discontinued owing to financial reasons. The onset of acute pancreatitis can be considered an adverse effect of PPFC. Although the association between PPFC administration and the onset of acute pancreatitis could not be clarified in this case, the onset of acute pancreatitis may have been associated with a decrease in liver function and/or increase in the false activity of lipase. PPFC has been considered highly safe in dogs, although care should be taken when administering medications to dogs suspected of having an underlying disorder.

Diagnostic utility of measuring lactate dehydrogenase levels and its isoenzyme activities for the evaluation of malignancy in feline pleural effusion and ascitic fluid.

Background: Lactate dehydrogenase (LDH) isoenzymes may be useful in the differential diagnosis of pleural effusion (PE) and ascitic fluid (AF) etiologies in cats since tissue damage induces their release, changing the pattern of their activity. Aim: This study aimed to determine the diagnostic utility of measuring LDH levels and isoenzyme activities in PE or AF in cats with malignancy. Methods: LDH levels and isoenzyme activities in the serum, PE, and AF were compared among cats in the malignant, infectious, and non-malignant, non-infectious groups. A receiver operating characteristic (ROC) analysis was performed to assess the accuracy in diagnosing feline malignancy. Results: Significant differences in LDH level and LDH isoenzyme activities in the PE and AF were observed among the three groups. The combination of LDH level and LDH-1 activity in PE or AF had the highest area under the ROC (AUC) values for discriminating malignant effusion from non-malignant effusion. The AUC of the combination of LDH level and LDH-1 activity in PE or AF was 0.874. The sensitivity and specificity of using the combination of LDH level (cut-off: <2,269 U/l) and LDH-1 activity (cut-off: <4.8%) in PE or AF for predicting malignancy with the highest AUC value were 94.4% and 72.7%, respectively. Conclusion: Our results suggest that the combination of LDH level and LDH-1 activity in PE or AF is a potential factor for diagnosing malignancy. Considering that LDH isoenzymes can be measured inexpensively and easily, LDH tests can be readily accommodated in veterinary clinical practice.

Subclinical wandering spleen in a cat with gastrointestinal lymphoma.

We accidentally detected a subclinical wandering spleen on preoperative ultrasonography in a cat with gastrointestinal lymphoma. If the spleen is not in the normal position, the wandering spleen should be considered.

Obese status is associated with accelerated DNA methylation change in peripheral blood of senior dogs.

Obesity and its associated comorbidities constitute a major and growing health problem worldwide not only involved with people but also dogs and cats. Although few genetic mutations have been associated with obesity in dogs, molecular mechanism remains to be clearly understood. Given the fact that DNA methylation leads to gene expression variability and has plasticity affected by metabolic phenotypes such as obesity in human, the objective of this study is to identify obesity-associated differentially methylated cytosine-phosphate-guanine (CpG) dinucleotide sites in dogs. With genome-wide DNA methylation analysis using next-generation sequencing for blood samples from fourteen Miniature dachshunds with body condition score (BCS) 4-5 and BCS ≥6, over 100,000 sites could be analysed to identify genomic locations of differentially methylated CpG sites. As a result, 191 differentially methylated CpG sites (89 CpG sites were hypermethylated in BCS ≥6 and 102 were hypermethylated in BCS 4-5) were identified. These sites included promoter regions of Kisspeptin receptor (KISS1R) and Calcyphosine 2 (CAPS2) genes which were subsequently validated by bisulfite-pyrosequencing for another set of 157 dog blood samples. KISS1R methylation levels were found to be higher in BCS ≥6 group than BCS 4-5 in senior (>84 months) dogs. Especially male dogs but not female dogs as well as uncastrated male dogs but not castrated male dogs showed this trend. DNA methylation of KISS1R gene will be useful for understanding of comprehensive epigenetic change in obese dogs.

Prognostic utility of preoperative neutrophil-lymphocyte ratio in cats with malignant mammary tumors.

Although studies have determined the neutrophil-lymphocyte ratio (NLR) to be a useful prognostic predictor in various human cancers, little is known regarding its preoperative prognostic significance in feline mammary tumors (FMTs). Thus, the present study aimed to determine the prognostic utility of preoperative peripheral blood NLR in malignant FMTs. Accordingly, medical records of 34 cats that had undergone mastectomy were searched, after which those histopathologically confirmed to have malignant FMTs were included. Univariate analysis was used to determine the effect of each variable on one-year survival time. Factors determined to be significant during univariate analysis were subsequently included for multivariate analysis. Receiver operating characteristic analysis was used to assess the accuracy of the tests for predicting one-year survival. Univariate analysis showed that a high NLR and large tumor size were significantly associated with shorter survival times (P < 0.01, for both). No significant differences in other factors were observed. Multivariate analysis showed that a high NLR (hazard ratio, 25.03; 95% confidence interval, 2.98-210.40; P < 0.01), but not tumor size and age, was an independent prognostic factor for poor outcomes. The sensitivity and specificity of the NLR (cut-off, 5.67) for predicting one-year survival were 93.8% and 88.9%, respectively. This study suggests that the NLR could potentially be used as a preoperative prognostic factor in FMTs. Preoperative NLR may help to establish therapeutic modalities by allowing prognostic evaluation among cats with mammary tumors.

Primary Adenosquamous Cell Carcinoma of the Ileum in a Dog.

A 9-year-old male, castrated Chihuahua was examined because of a 7-day history of intermittent vomiting. A mass in the small intestine was identified on abdominal radiography and ultrasonography. Laparotomy revealed a mass lesion originating in the ileum, and surgical resection was performed. The mass was histologically diagnosed as adenosquamous cell carcinoma. Chemotherapy with carboplatin was initiated, but the dog was suspected to have experienced recurrence 13 months after surgery and died 3 months later. To our knowledge, this is the first case report to describe the clinical course of adenosquamous cell carcinoma in the small intestine of a dog.

A Clinical Investigation on Serum Amyloid A Concentration in Client-Owned Healthy and Diseased Cats in a Primary Care Animal Hospital.

Although measurement of serum amyloid A (SAA) concentration in client-owned cats has already been shown to be clinically useful, limited data are available on common diseases at primary care hospitals. In this study, we measured the SAA concentration in cats with various diseases and investigated their clinical significance using a primary care hospital as a population. We measured the SAA concentrations in healthy cats (n = 98) and those with various clinical signs (n = 444). The SAA concentrations in healthy cats did not differ significantly by age, breed, sex, and presence/absence of neutering/spaying. The SAA concentrations were significantly higher in the diseased cat group than in the healthy cat group (p < 0.001). We observed significant increases in SAA concentrations in cats with confirmed diagnosis of inflammatory disease such as upper respiratory tract infections (p < 0.001), pneumonia (p < 0.001), pyometra (p = 0.001), and feline infectious peritonitis (p < 0.001), compared with those observed in healthy cats. Conversely, no increase was observed in cardiomyopathy, hyperthyroidism, and diabetes mellitus without systemic inflammation. In univariate analysis, survival at 30 days (p = 0.03) differed significantly between the low and high SAA concentration groups, but not at 180 days. In multivariate analysis, survival at 30 days did not significantly affect SAA concentration. Measurement of SAA concentration is a useful biomarker for detecting the presence or absence of inflammation in diseased cats. However, it may not be useful as a biomarker for determining the prognosis of the disease.

Evaluation of atrial natriuretic peptide and cardiac troponin I concentrations for assessment of disease severity in dogs with naturally occurring mitral valve disease.

OBJECTIVE: To evaluate and compare the clinical usefulness of plasma atrial natriuretic peptide (ANP) and cardiac troponin-I (cTnI) concentrations for assessment of disease severity in dogs with naturally occurring mitral valve disease (MVD). ANIMALS: 316 dogs with MVD and 40 healthy control dogs. PROCEDURES: Each dog underwent a physical examination and echocardiographic and thoracic radiographic assessments. Blood samples were obtained and processed for measurement of plasma ANP and cTnI concentrations. Dogs with MVD were categorized into 3 groups (stages B1 [no clinical signs or evidence of cardiac enlargement], B2 [no clinical signs with evidence of cardiac enlargement], and C [history of congestive heart failure and pulmonary edema]) on the basis of American College of Veterinary Internal Medicine guidelines. Receiver operating characteristic curve analysis was used to evaluate the accuracy of plasma ANP and cTnI concentrations for assessment of MVD severity. RESULTS: Plasma ANP and cTnI concentrations increased as disease severity increased. Median plasma ANP concentrations for all 3 MVD groups and median plasma cTnI concentrations for the stage B2 and C groups were significantly greater than the corresponding concentrations for the control group. Plasma ANP concentration, but not cTnI concentration, appeared to be useful for detection of dogs with subclinical (stages B1 and B2) MVD, whereas both concentrations appeared useful for detection of dogs with stage C MVD. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that plasma ANP and cTnI concentrations should not be used independently to diagnose MVD but can be used to assess MVD severity and supplement echocardiographic findings.

Resolution of reactive arthritis after splenectomy in a dog with a nonneoplastic splenic hyperplastic nodule.

CASE DESCRIPTION: A 10-year-old spayed female Golden Retriever was examined because of a 3-month history of lethargy, anorexia, and stumbling gait. CLINICAL FINDINGS: A splenic mass was identified on abdominal radiography and ultrasonography, and results of clinicopathologic findings indicated nonregenerative anemia, leukocytosis, and high serum C-reactive protein (CRP) concentration. To further investigate the cause of the dog's high serum CRP concentration, radiography and arthrocentesis were performed bilaterally on the carpal and stifle joints. On the basis of results, anemia of chronic disease associated with polyarthritis caused by the localized splenic mass was suspected. TREATMENT AND OUTCOME: After splenectomy, there were improvements in the dog's clinical signs, polyarthritis, nonregenerative anemia, and serum CRP concentration. The splenic mass was histologically diagnosed as a nonneoplastic splenic hyperplastic nodule with evidence of omental adhesion. CLINICAL RELEVANCE: Findings indicated that nonneoplastic splenic hyperplastic nodules could result in reactive polyarthritis, although such nodules have not to our knowledge been described previously as an underlying cause of polyarthritis. Therefore, veterinarians should investigate for nonneoplastic splenic hyperplastic nodules in addition to other typical underlying causes when treating dogs with polyarthritis.

Complete Remission of Associative Immune-Mediated Hemolytic Anemia in a Dog Following Surgical Resection of Intestinal Leiomyosarcoma.

A twelve-year-old male castrated Chihuahua with a severe, microcytic, hypochromic, and nonregenerative direct antiglobulin test positive anemia characterized by marked spherocytosis was referred to the veterinary hospital. Abdominal ultrasound revealed a peritoneal mass of unclear origin. Transfusion, followed by mass resection, rapidly resolved the anemia without further immunosuppressive treatment. Histopathology confirmed extraluminal jejunal leiomyosarcoma. Multiple mechanisms, including immune-mediated destruction, likely contributed to the anemia. To the authors' knowledge, this is the first report that describes the resolution of immune-mediated hemolysis in a dog after the removal of an intestinal neoplasm.

Investigation of serum cortisol concentration as a potential prognostic marker in hospitalized dogs: a prospective observational study in a primary care animal hospital.

BACKGROUND: Dogs with various medical diseases are usually treated at hospitals; however, the prognostic markers in dogs remain unknown. The aim of this study was to investigate the ability of serum cortisol concentration (SCC) to predict the prognosis of dogs with medical diseases. At 0 and 24 h after hospitalization, the neutrophil count, lymphocyte count, blood glucose concentration, and SCC were measured. Survival for 30 days from the time of hospitalization was investigated, and the dogs were divided into a survivor group and a non-survivor group. RESULTS: The neutrophil count at 24 h, SCC at 24 h, increase in SCC from 0 to 24 h (Inc-SCC), and the rate of increase in SCC from 0 to 24 h (R-Inc-SCC) were significantly higher in the non-survivor group than in the survivor group. The area under the receiver operating characteristic (ROC) curve values for the neutrophil count at 24 h, SCC at 24 h, Inc-SCC, and R-Inc-SCC were 0.695, 0.72, 0.63, and 0.66, respectively. Using the highest area under the ROC curve value, the sensitivity and specificity of SCC at a cutoff level of 6.6 µg/dL for predicting mortality were 89.5 and 61.9%, respectively. Moreover, the Kaplan-Meier curves confirmed the significant prognostic influence of SCC at 24 h. CONCLUSIONS: SCC as a marker of stress is a useful biomarker for predicting the prognosis of dogs with medical diseases requiring hospital treatment.

Diagnostic utility of cardiac troponin I in cats with hypertrophic cardiomyopathy.

BACKGROUND: Cardiac troponin I (cTnI) is useful for assessing hypertrophic cardiomyopathy (HCM) in cats. OBJECTIVE: To measure plasma cTnI concentrations in healthy cats and evaluate the clinical utility of cTnI in determining the severity of HCM. ANIMALS: Clinically healthy cats (n = 88) and cats with HCM (n = 93). METHODS: Multicenter prospective study. Cats with HCM, including hypertrophic obstructive cardiomyopathy at various stages, were diagnosed using echocardiography. Plasma cTnI concentrations were analyzed by a commercial laboratory. Receiver-operating characteristic curve analysis was used to evaluate the accuracy of plasma cTnI concentrations to detect HCM. RESULTS: The median cTnI concentration was 0.027 ng/mL (interquartile range, 0.012-0.048 ng/mL) in healthy cats. Concentrations were significantly higher in diseased cats than in healthy controls, and concentrations were significantly higher in cats with heart failure than in asymptomatic cats. A plasma cTnI concentration of 0.163 ng/mL had a sensitivity of 62.0% and specificity of 100% when used to distinguish normal cats from asymptomatic HCM cats without left atrial dilatation. A cutoff of 0.234 ng/mL had high sensitivity (95.0%) and specificity (77.8%) for assessing heart failure. The areas under the receiver-operating characteristic curves were 0.85 and 0.93, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Increased cTnI concentrations reflect the severity of HCM. If other causes of cardiac injury are ruled out, plasma cTnI concentration may be useful for predicting the severity of HCM in cats.

Pyoderma gangrenosum after trauma in a dog.

A 12-year-old male entire Miniature Pinscher presented with excoriations at various body sites, progressively forming ulcers and enlarging until arrested by treatment. Based on the clinical presentation and histopathological analyses, sterile neutrophilic dermatosis was suspected. Therefore, the dog was started on prednisolone. Marked improvement was achieved with prednisolone treatment, suggesting a diagnosis of pyoderma gangrenosum (PG). Transcription levels of cytokine mRNA in lesional skin before and after treatment from this dog were quantified by real-time RT-PCR. Transcription levels of tumor necrosis factor-α, interleukin (IL)-1ß, IL-8 and IL-17A were higher in lesional skin before treatment than after treatment. Levels of various cytokines could be increased in lesional skin of dogs with PG as well as in human patients with PG.

Long-term outcome of myotonia associated with hyperadrenocorticism in 2 dogs.

Two dogs were diagnosed with myotonia associated with hyperadrenocorticism and treated with trilostane. One dog showed temporary improvement, but the other dog showed no improvement. The dogs survived 2383 and 1902 days, respectively. Findings suggest that myotonia persists despite treatment, but this condition is not associated with a poor prognosis for survival.

Résultat à long terme de la myotonie associée à l'hyperadrénocorticisme chez 2 chiens. Deux chiens ont été diagnostiqués avec la myotonie associée à l'hyperadrénocorticisme et traités avec du trilostane. Un chien a manifesté une amélioration temporaire, mais l'autre n'a manifesté aucune amélioration. Les chiens ont survécu 2383 et 1902 jours, respectivement. Les résultats ont suggéré que la myotonie persiste malgré le traitement, mais cette affection n'est pas associée à un pronostic défavorable pour la survie.(Traduit par Isabelle Vallières).

Parathyroid hormone-related protein-induced hypercalcemia due to osteosarcoma in a cat.

A 15-year-old castrated male mixed-breed cat was presented with a history of sarcoma of the distal right hind limb. Biochemical analysis revealed increased concentrations of blood urea, creatinine, total calcium, ionized calcium, and parathyroid hormone-related protein (PTHrP). The mass was removed surgically by amputation of the hind limb. Osteosarcoma was diagnosed based on histopathologic examination. All abnormal serum analyte concentrations improved immediately after surgery, including azotemia, total calcium, ionized calcium, and PTHrP. The biochemical results were attributed to osteosarcoma causing PTHrP-induced hypercalcemia.

Markedly increased expression of interleukin-8 in the colorectal mucosa of inflammatory colorectal polyps in miniature dachshunds.

Inflammatory colorectal polyps (ICRPs) in miniature dachshunds were recently recognized as a major cause of large bowel diarrhea in this dog breed in Japan. ICRPs are characterized by the formation of multiple small polyps and/or space-occupying large polyps in the colorectal area and are thought to be a novel form of inflammatory bowel disease (IBD). To explore key mediators in the pathogenesis of ICRPs, we analyzed several pro-inflammatory cytokine (IL-1ß, IL-6, TNF-α, IL-8, IL-12p35, IL-12/23p40, and IL-23p19) mRNA expressions in colorectal polyps in ICRP dogs by quantitative PCR. Among these cytokines, IL-8 mRNA expression was markedly up-regulated in large polyps. To examine IL-8 protein expression, we analyzed IL-8 protein level and its location in colorectal mucosal specimens of ICRP dogs by ELISA and immunofluorescence microscopy. IL-8 protein was significantly increased in large polyps and serum in dogs with ICRPs compared to controls. By immunofluorescence microscopy, IL-8 was only localized in macrophages, but not in mucosal epithelial cells or neutrophils. IL-8-positive macrophages were significantly increased in large polyps compared to controls. These results suggest that IL-8 is produced mainly by macrophages and may induce neutrophil infiltration in the colorectal area of ICRP dogs.

Expression of CD4+ T cell cytokine genes in the colorectal mucosa of inflammatory colorectal polyps in miniature dachshunds.

Inflammatory colorectal polyps (ICRPs) in miniature dachshunds are recently recognized as a major cause of large bowel diarrhea in this dog breed in Japan. ICRPs are characterized by the formation of multiple small polyps and a space-occupying large polyp in the colorectal area, and are thought to be a novel form of inflammatory bowel disease (IBD). In humans, specific cytokine patterns attributed to T helper (Th)1, Th17 and regulatory T cells have important roles in the pathogenesis of IBD. Thus, the aim of the present study was to assess the gene expression of cytokines of T cell subsets in the colorectal mucosa from dogs with ICRPs. Colorectal mucosal specimens from 10 dogs with ICRPs and 14 control dogs were used in this study. Interferon (IFN)-γ, interleukin (IL)-4, IL-17A and IL-10 mRNA expression was assessed using quantitative real-time PCR. IL-17A mRNA expression was significantly increased in large polyps compared to small polyps and controls. IFN-γ and IL-10 mRNA expression in large polyps were significantly higher than in controls. There was no significant difference in IL-4 mRNA expression among the three groups. IL-17A is thought to play important roles in the pathogenesis of ICRPs. IL-10 up-regulation could oppose the proinflammatory function of IL-17A.

Mastitis caused by Mycobacterium kansasii infection in a dog.

A 2-year, 7-month-old female Chihuahua was admitted for a mammary mass measuring one cm in diameter. The dog had a history of demodicosis for 4 months and showed signs of pseudopregnancy at the time of the visit. Cytologic examination of an aspirate of the mass revealed a large number of macrophages containing nonstaining bacterial rods, which were acid-fast in a Ziehl-Neelsen stain, suggesting mycobacterial infection. Histologic examination of the mass revealed a pyogranulomatous mastitis characterized by an infiltration with macrophages containing acid-fast bacteria. Mycobacterium kansasii was subsequently cultured and identified by PCR. Surgical excision of the mass resulted in the growth of other dermal masses, but antimycobacterial treatment with rifampin and clarithromycin resolved these masses within 1 month. Three months after discontinuation of the treatment, similar organisms were found in aspirates of the enlarged bilateral inguinal lymph nodes by cytologic examination. Despite antimycobacterial treatment for another 4 months, there was no improvement and demodicosis also recurred. The dog eventually died of lymphoma 5 months after the relapse of mycobacterial infection. Although M kansasii is considered an important pathogen for pulmonary and cutaneous disease in people, there is only one report in a dog with an infection in a pleural effusion. As both adult-onset demodicosis in dogs as well as mycobacterial infection in people have been associated with T-lymphocyte deficiency, the M kansasii infection in this dog may have been associated with a condition of immune compromise.