Veterinary clinical pathology最新文献

Background: Pleural effusion (PE) in cats can result from transudative or exudative processes. Transudates are caused by decreased colloid osmotic pressure (↓COP) or elevated hydrostatic pressure (↑HP) gradient, while exudates arise from increased pleural capillary permeability. Diagnostic classification approaches traditionally rely on pleural effusion total protein (TP_PE) and total nucleated cell counts (TNCC_PE). In contrast, Light's criteria employing pleural effusion lactate dehydrogenase (LDH_PE), LDH_PE divided by serum LDH (LDH_ratio), and pleural effusion to serum protein ratio (TP_ratio) are more accurate than classification based on TP_PE/TNCC_PE in humans and show promise in cats.

Objectives: To assess the diagnostic accuracy of TNCC_PE, LDH_PE, LDH_ratio, TP_PE, and TP_ratio in classifying feline PEs in ↑HP-transudates, ↓COP-transudates, or exudates and to compare simplified Light's criteria (which relies solely on LDH_PE) with Light's criteria and existing veterinary classification schemes based on TP_PE and TNCC_PE (named Vet-A and Vet-B).

Methods: Cross-sectional study including 83 client-owned cats with PE.

Results: There were 55 exudates, 28 ↑HP-transudates, and 0 ↓COP-transudates. All the variables analyzed were significantly different between exudates and transudates. Simplified Light's criteria correctly classified 50/55 exudates and 26/28 ↑HP-transudates (sensitivity = 91%, specificity = 93%, accuracy = 92%). Light's criteria correctly identified 55/55 exudates but misclassified 15/28 ↑HP-transudates as exudates (accuracy = 82%). Traditional veterinary schemes showed lower accuracies: Vet-A (57%) and Vet-B (74%). Simplified Light's criteria outperformed Vet-A (p < 0.001) and Vet-B (p = 0.007) and trended higher than Light's criteria (p = 0.096).

Conclusions: Simplified Light's criteria demonstrate excellent diagnostic accuracy, outperforming traditional veterinary classification schemes.

Background: microRNAs (miRNA) have been proposed as biomarkers for pancreatitis in dogs due to their high peak concentrations and experimental correlation with acinar cell injury. However, analytical validation and the effect of interfering substances are unknown.

Objective: The study aimed to (i) develop and analyze the analytical validity of an assay for miRNA-148a and miRNA-375 in serum and (ii) compare miRNA-148a and miRNA-375 between healthy and pancreatitis-affected dogs.

Methods: Circulating miRNAs were quantified from serum of healthy (n = 40) and pancreatitis-affected dogs (n = 40). Reference intervals were established, and serum miRNAs were compared between groups. Linearity was assessed via dilutional parallelism (observed/expected ratio, O/E). Precision was assessed via intra- and inter-assay coefficients of variation (% CV). Intralipid, bilirubin, and hemoglobin were tested as interferants.

Results: For miRNA-148a, the RI was < 33 000 gene copies and mean O/E was 100.4%. Mean inter-assay and intra-assay % CV were 1.5% and 1.3% respectively. For miRNA-375, the RI was < 4 gene copies and mean O/E was 100.1%. Mean inter-assay and intra-assay % CV were 3.2% and 0.5% respectively. Hemolysis led to false increases in both assays (p = 0.003 - < 0.0001). miRNA-148a was reduced in dogs with pancreatitis (mean 192 gene copies) compared with healthy controls (mean 2760 gene copies; p < 0.0001), but miRNA-375 was not different between groups (p = 0.94).

Conclusions: The miRNA assays were analytically valid; however, hemolysis, a common interferant in pancreatitis cases, may impact test results. miRNA-148a was suppressed in dogs with pancreatitis. It is unclear whether this represents a cause or a consequence of the disease.

Backgrounds: Studies investigating the influence of sampling methods on point-of-care viscoelastic test (VCM Vet) results are limited.

Objectives: Investigating the impact of blood sampling methods on VCM Vet results in dogs, and determining if results are affected by hematological parameters, and blood sampling difficulty.

Methods: Two VCM assays were performed on 52 healthy dogs. Blood sample was first collected from a direct jugular venipuncture on all dogs to run a baseline VCM Vet assay, perform a CBC, and measure fibrinogen concentration. A second VCM Vet assay was performed one hour later with blood sampling methods randomized as follows: contralateral jugular using a vacutainer, direct stick in a saphenous vein, or blood sampling via a cephalic intravenous catheter.

Results: Reference intervals were established for each VCM Vet parameter with the first blood sample. The intra-class correlation (ICC) between sampling methods was poor (< 0.5). There was a weak positive correlation between hematocrit and CT (p < 0.042), a weak negative correlation between platelet count and CFT (p < 0.01), and a weak positive correlation between platelet count and alpha (p = 0.002), A10 (p < 0.001), A20 (p < 0.001), and MCF (p < 0.001).

Conclusions: Sampling protocols influence VCM Vet results. Each sampling method is reliable but not correlated. Follow-up on a patient should be performed using the same sampling method and site. CBC results should be known before interpreting results.

Artificial intelligence (AI), particularly through machine learning and deep learning, presents opportunities for the enhancement of the workflow of the veterinary clinical pathologist. This review introduces basic concepts in AI in a nontechnical manner and explores the qualification and integration of AI in veterinary clinical pathology. The veterinary clinical pathologist must play an active role in defining the intended use, design, and qualification of these methods as well as the plan for monitoring their responsible application in practice.

Background: An 8.8-year-old male neutered Persian cat was presented with a history of recent intermittent mild lethargy, hyporexia, and weight loss. Physical examination revealed pale mucous membranes, an enlarged right mandibular lymph node, and organomegaly suspected to be an enlarged spleen. A CBC showed an extreme leukocytosis composed almost exclusively of small lymphocytes and a moderate, microcytic, slightly regenerative anemia.

Objectives: We aimed to diagnose the cause of the extreme leukocytosis, to classify the circulating leukocytes, and to determine the response to therapy.

Methods: Blood film review, biochemical and iron analysis, abdominal ultrasound, flow cytometric evaluation of leukocytes, serum and urine protein electrophoresis, and polymerase chain reaction for antigen receptor rearrangement (PARR) were performed.

Results: Leukocytes were mostly small lymphocytes with scant pale basophilic cytoplasm, and round or convoluted nuclei with coarse to finely clumped chromatin. There was hyperglobulinemia and iron deficiency. Imaging showed splenomegaly and multiple enlarged abdominal lymph nodes. On flow cytometry, the lymphocytes were positive for CD18, CD21, and MHC II, consistent with B-cell lymphocytic leukemia. The cat had mild hyperglobulinemia and 2+ proteinuria, and serum electrophoresis results were interpreted as a polyclonal gammopathy. PARR showed clonal rearrangement of IGH2 and IGH3 loci. Treatment with oral prednisolone and chlorambucil resulted in resolution of the clinical signs and reduction in the leukocytosis.

Conclusions: B-cell chronic lymphocytic leukemia (B-CLL) was diagnosed, which in cats is much less frequent than T-CLL. There was a favorable response to alkylating therapy, and the cat survived for 19 months.

A 12-year-old female spayed Border Collie dog was presented for evaluation of 6 months of intermittent hematuria and weight loss. A highly vascularized right renal mass deforming the renal architecture and paraneoplastic hypertrophic osteopathy were found. Cytologic evaluation of the mass obtained by fine-needle aspiration guided by ultrasound revealed mesenchymal cells with a moderate amount of bluish cytoplasm, moderately defined cell borders, and spindle to stellate or roundish morphology. The nuclei were centrally located, with a coarse chromatin pattern, round to oval, and occasionally bean-shaped. Usually, a single distinct nucleolus per nucleus with minimal size variation was noted. Anisocytosis and anisokaryosis were moderate. The cytologic interpretation was mesenchymal proliferation with moderate atypia, most consistent with soft tissue sarcoma. Right ureteronephrectomy was performed. Histologic evaluation showed a neoplastic proliferation located beneath the lamina propria of the transitional epithelium of the renal pelvis and infiltrating the renal medulla. Immunohistochemistry for protein S-100, laminin, and desmin was performed to further characterize the lesion as a nerve sheath tumor. The hematuria disappeared 4 days after the surgery; 5 months later, no alterations were observed in the general examination, and the paraneoplastic hypertrophic osteopathy mildly improved. This is the first cytologic description of a primary renal malignant nerve sheath tumor in the renal pelvis of a dog with paraneoplastic hypertrophic osteopathy.

Background: Point-of-care testing (POC) is widely utilized for rapid results for many different analytes. A new feline-specific N-terminal pro-brain natriuretic peptide (NT-proBNP) quantitative assay (Vcheck V200, Bionote Inc) is currently available but has not undergone independent validation.

Objective: To validate the Vcheck POC quantitative assay for feline NT-proBNP.

Methods: Validation was performed in accordance with the American Society for Veterinary Clinical Pathology guidelines utilizing serum samples from 62 cats. Precision was determined for low (50-100 pmol/L), mid (101-300 pmol/L), and high (> 301 pmol/L) pools short-term (20 repetitions) and long-term (5 repetitions each day for 5 days). Linearity, methods comparison, interference testing, and sample stability were evaluated.

Results: The within-day coefficients of variability (CV) were low pool = 12.6%, mid pool = 10.4%, and high pool = 8.7%. The within-week CV was low pool = 9.9%, mid pool = 14.9%, and high pool = 6.9%. The assay was linear over the analytical range of 53-1488 pmol/L (R² = 0.99, p < 0.0001). Paired samples between the feline Cardiopet NT-proBNP (IDEXX) and Vcheck assays demonstrated a mean difference of 11 pmol/L (2.3%), p = 0.38, between assays with minimal constant or proportional bias. Hemolysis and lipemia did not affect assay performance, while all icteric samples were invalid. Significantly lower values were identified in samples after 2 and 4 h when stored at 20°C and 4°C, respectively.

Conclusions: The Vcheck V200 has acceptable precision, accuracy, and compares favorably with commercially available assays and is a viable POC quantitative assay for feline NT-proBNP.

Digital cytopathology is increasingly used in veterinary medicine, yet standardized quality guidance for its safe and effective implementation has been lacking. As more reference laboratories and individual clinics adopt this technology, challenges related to sample preparation, staining variability, scanner capabilities, software design, operator training, and limitations in optical resolution have emerged. To address these issues, the ASVCP Quality Assurance and Laboratory Standards Committee developed comprehensive guidelines for the preanalytical, analytical, and postanalytical phases of digital cytologic evaluation, with the aim of enhancing diagnostic accuracy, improving workflow efficiency, and supporting safer clinical decision-making. The target audience is veterinary pathologists, residents, laboratory personnel, and practices currently using or planning to adopt digital cytopathology. Key recommendations include ensuring appropriate sample preparation and placement to support accurate digital capture; selecting and validating scanners and software with performance characteristics appropriate for cytologic specimens; implementing standardized training for operators and pathologists; and establishing procedures for assessing digital image quality, managing incomplete or low-quality scans, and determining when glass slide review is required. Because digital cytopathology continues to evolve rapidly and the current evidence base remains limited, these guidelines are intended as a minimum standard and should be revisited within five years to incorporate emerging QA data and technological advancements.

A 5-year-old LaMancha goat buck presented for evaluation of a 1-week history of progressive lameness and carpal swelling in the left thoracic limb. Cytology performed on an aspirate from the left radiocarpal joint identified abundant round to oval, thickly encapsulated, extracellular yeast organisms that occasionally exhibited narrow-based budding, features compatible with Cryptococcus spp. yeast. The patient was euthanized due to poor prognosis, and a necropsy was performed. Histopathology confirmed the presence of numerous fungal yeast associated with the left radiocarpal joint. A cryptococcal serum latex agglutination test was negative. However, panfungal PCR and fungal culture identified the organisms as Cryptococcus gattii. This report describes, for the first time, a case of a C. gattii infection affecting the joint of a goat.

Acute myeloid leukemia (AML) poses significant challenges in veterinary medicine, with limited treatment options and poor survival rates. While substantial progress has been made in characterizing human AML, translating these advancements to veterinary practice has been hindered by limited molecular understanding and diagnostic tools. The case study presented illustrates the application of whole genome sequencing in diagnosing AML in a dog, showcasing its potential in veterinary oncology. Our approach facilitated comprehensive genomic analysis, identifying mutations in genes that may be associated with AML pathogenesis in dogs, such as KRAS, IKZF1, and RUNX1. However, without supportive evidence of its clinical utility (eg, association with response to treatment or prognosis), the information is limited to exploration. This article reviews the comparative features of canine AML with human AML and discusses strategies to shrink the knowledge gap between human and veterinary medicine with cost-effective next-generation sequencing (NGS) techniques. By utilizing these approaches, the unique and shared molecular features with human AML can be identified, aiding in molecular classification and therapeutic development for both species. Despite the promise of NGS, challenges exist in implementing it into routine veterinary diagnostics. Cost considerations, turnaround times, and the need for robust bioinformatics pipelines and quality control measures must be addressed. Most importantly, analytical and clinical validation processes are essential to ensure the reliability and clinical utility of NGS-based assays. Overall, integrating NGS technologies into veterinary oncology holds great potential for advancing our understanding of AML and improving disease stratification, in hopes of improving clinical outcomes.