Guidelines for resident training in veterinary clinical pathology. IV: Laboratory quality management-Teaching domains, competencies, and suggested learning outcomes.

BACKGROUND: The 2019 ASVCP Education Committee Forum for Discussion, presented at the annual ASVCP/ACVP meeting, identified a need to develop recommendations for teaching laboratory quality management principles in veterinary clinical pathology residency training programs. OBJECTIVES: To present a competency-based framework for teaching laboratory quality management principles in veterinary clinical pathology residency training programs, including entrustable professional activities (EPAs), domains of competence, individual competencies, and learning outcomes. METHODS: A joint subcommittee of the ASVCP Quality Assurance and Laboratory Standards (QALS) and Education Committees executed this project. A draft guideline version was reviewed by the ASVCP membership and shared with selected ACVP committees in early 2022, and a final version was voted upon by the full QALS and Education Committees in late 2022. RESULTS: Eleven domains of competence with relevant individual competencies were identified. In addition, suggested learning outcomes and resource lists were developed. Domains and individual competencies were mapped to six EPAs. CONCLUSIONS: This guideline presents a framework for teaching principles of laboratory quality management in veterinary clinical pathology residency training programs and was designed to be comprehensive yet practical. Guidance on pedagogical terms and possible routes of implementation are included. Recommendations herein aim to improve and support resident training but may require gradual implementation, as programs phase in necessary expertise and resources. Future directions include the development of learning milestones and assessments and consideration of how recommendations intersect with the American College of Veterinary Pathologists training program accreditation and certifying examination.

Lymphopenia predicts reduced survival in canine hepatocellular carcinoma.

Platelet to lymphocyte ratio (PLR) is a prognostic marker in human hepatocellular carcinoma (HCC) however, its utility in canine HCC has not been explored. The aim of the study was to determine if PLR could predict survival outcomes in 42 dogs with HCC. PLR was not a significant predictive factor (p = 0.15) but lymphopenia alone was significantly correlated with a reduced probability of survival (p = 0.024). Further studies are needed to evaluate if peripheral lymphocyte count mirrors that of the tumor microenvironment in canine HCC.

Prevalence of AO blood group and level of agreement for AO blood-typing methods in pet pigs from Louisiana.

OBJECTIVES: Pet pigs are a species of growing medical interest, and evidence-based practices for blood transfusions are needed. The objectives of this study were to quantify the prevalence of 3 blood group (Bg) phenotypes ("A" and "Aweak " resulting from EAAAA and EAAAO , "0" from EAA00 , or "-" from EAA00 or SSS alleles) in pet pigs and compare results using a human blood-typing card (EldonCard), standard saline agglutination (SSA), and polymerase chain reaction (PCR) sequencing. DESIGN: Cross-sectional study. SETTING: University veterinary teaching hospital. ANIMALS: Ninety-seven pet pigs from Louisiana. INTERVENTIONS: Blood was sampled from randomly selected pet pigs of various breeds, anticoagulated with EDTA, and typed using each investigated test according to the manufacturers' directions or standard laboratory technique. Samples for PCR analysis were stored at -80°C until analysis. Phenotypes were screened via EldonCard. Association between Bg and sex was investigated using chi-square test, with significance at P < 0.05. Kappa (κ) statistic was used to measure the level of agreement between the 3 tests. MEASUREMENTS AND MAIN RESULTS: Pot-bellied pigs represented the majority (84.5%) of this population, with 52 (53.6%) males and 45 (46.4%) females. Genotypic frequencies were 30%, 30%, and 40% for "EAAAA ," "EAAAO ," and "EAA00 ," respectively. Based on EldonCard, 38 phenotypes were classified as "A," 5 as "Aweak ," and 54 as "0" or "-." Results were identical for Bg, with the 3 tested techniques in 90% (45/50) of samples. Agreement between EldonCard and PCR was almost perfect (49/50 [98%], κ = 0.959; P < 0.001). Agreement between SSA and PCR, and EldonCard and SSA was substantial (46/50 [92%], κ = 0.803, P < 0.001 and 93/97 [95.9%], κ = 0.764, P < 0.001, respectively). CONCLUSIONS: The most common blood type was "0" or "-" (55.7%), followed by "A" (39.2%) and "Aweak " (5.1%). There was strong agreement between EldonCard and PCR testing. EldonCard allowed for rapid and reliable phenotype identification ("A," "Aweak ," and "0" or "-") and represents a clinically applicable laboratory method for blood typing in pet pigs.

Cytomorphology of Deep Mycoses in Dogs and Cats.

The purpose of this review is to familiarize clinical pathologists and clinicians with the cytomorphologic features associated with deep mycoses in dogs and cats. The goals are to develop a more unified approach to the description and interpretation of fungal cytomorphology and to facilitate the categorization of fungi that do not produce unique morphologic structures in tissue.

Hyphema and secondary glaucoma as a presenting complaint in a dog with chronic lymphocytic leukemia.

A 5-year-old Pomeranian was diagnosed with anterior uveitis, hyphema, and secondary glaucoma OD. Concurrent retinal hemorrhage, perivascular sheathing, and papilledema were identified OS. Work-up identified small cell lymphocytosis (>900 × 109/L), anemia, and thrombocytopenia. The patient was diagnosed with B-cell chronic lymphocytic leukemia as a cause of the ocular findings.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Canine memory T-cell subsets in health and disease.

A more complete understanding of canine T-lymphocyte immunity is necessary for improving diagnostic and therapeutic approaches to canine diseases, developing cell-based canine immunotherapeutics, and evaluating dogs as large mammal models for comparative immunology research. The aim of this study was to utilize CD45RA (indicating antigen inexperience) and CD62L (indicating lymph node homing capability), to quantify canine memory T-cell subsets in healthy dogs and dogs with various diseases. Peripheral blood mononuclear cells (PBMCs) were prospectively collected from dogs belonging to one of four groups:dermatologic inflammation (n = 9), solid tumors (n = 9), lymphoma (n = 9), and age-/weight-matched healthy control dogs (n = 15). Dogs receiving prednisone or any other immunomodulating medication within two weeks were excluded. Flow cytometry was performed and T-cell subsets were defined as CD4+ or CD8+, and naïve (TN), central memory (CM), effector memory (EM), or terminal effector memory re-expressing CD45RA (TEMRA). T-cell subset proportions were compared between each disease group and their healthy age-/weight-matched controls using a Mann-Whitney test. Significantly increased %CD8+ TN (P = 0.036) and decreased %CD8+ TEMRA (P = 0.045) were detected in dogs with dermatologic inflammation compared to healthy controls. Furthermore, %CD4+ TN positively correlated with Canine Atopic Dermatitis Extent and Severity Index (CADESI) score within the inflammation group (ρ = 0.817, P = 0.011). No significant differences between either cancer group and their healthy controls were detected. Taken together, these data indicate that dermatologic inflammation can alter proportions of peripheral blood T-cell subsets, possibly due to the migration of antigen-specific T-cells into tissues. Furthermore, these findings support the utility of CD45RA and CD62L in characterizing clinical canine immune responses.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Feline mesothelioma: case report and review of cytologic, immunocytochemical, histopathologic, and immunohistochemical findings.

Mesotheliomas are uncommon neoplasms that arise from mesothelial cells in either the abdominal or thoracic cavities and are rarely diagnosed in cats. A 10-y-old spayed female domestic shorthair cat was presented to the Louisiana State University oncology service for evaluation of a large amount of abdominal effusion. Abdominal ultrasound identified a large mesenteric mass with numerous ill-defined nodules. An abdominocentesis was performed with cytologic and immunocytochemical findings consistent with a neoplastic effusion, with large clusters of epithelioid cells that exhibited strong cytoplasmic expression of pancytokeratin, vimentin, and Wilms tumor 1 antigens. Further testing was declined, and meloxicam was prescribed until the cat died 23 d after initial presentation. Upon postmortem examination, the omentum was contracted into a firm mass adhered to multiple organs and accompanied by numerous small white nodules throughout the abdominal cavity. On histopathology and immunohistochemistry, neoplastic cells were found throughout the abdominal cavity; 60-95% exhibited moderate-to-strong cytoplasmic immunoreactivity for cytokeratin, vimentin, and Wilms tumor 1 protein. The final diagnosis was an epithelioid mesothelioma. Our case illustrates the utility of cytology, immunocytochemistry, and its relation to histology and immunohistochemistry. We also reviewed the reported cases of feline mesothelioma.

Development of and Validity Evidence for a Canine Ocular Model for Training Novice Veterinary Students to Perform a Fundic Examination.

Indirect fundoscopy is challenging for novice learners, as patients are often intolerant of the procedure, impeding development of proficiency. To address this, we developed a canine ocular simulator that we hypothesized would improve student learning compared to live dogs. Six board-certified veterinary ophthalmologists and 19 second-year veterinary students (novices) performed an indirect fundic examination on the model and live dog. Prior to assessment, novices were introduced to the skill with a standardized teaching protocol and practiced (without feedback) with either the model (n = 10) or live dog (n = 9) for 30 minutes. All participants evaluated realism and usefulness of the model using a Likert-type scale. Performance on the live dog and model was evaluated in all participants using time to completion of task, performance of fundic examination using a checklist and global score, identification of objects in the fundus of the model, and evaluation of time spent looking at the fundus of the model using eye tracking. Novices (trained on simulator or live dogs) were compared in fundic examination performance on the live dog and identification of shapes in the model. In general, experts performed the fundic examination faster (p ≤ .0003) and more proficiently than the novices, although there were no differences in eye tracking behavior between groups (p ≥ .06). No differences were detected between training on simulator versus live dog in development of fundoscopy skills in novices (p ≥ .20). These findings suggest that this canine model may be an effective tool to train students to perform fundoscopy.

Trapped Neutrophil Syndrome in a Border Collie.

A 10 wk old female border collie was presented for hemorrhagic diarrhea and pelvic limb lameness. Examination revealed pain and effusion in multiple appendicular joints and pyrexia. Clinicopathologic testing revealed moderate neutropenia as well as nondegenerate neutrophilic inflammation in multiple joints. Radiographs showed capsular joint swelling and heterogeneous metaphyseal lucencies in the distal radius, ulna, femur, and tibia. Genetic testing confirmed a mutation in the vacuolar protein sorting-associated protein 13B gene and a diagnosis of trapped neutrophil syndrome (TNS). Within 24 hr of initiating prednisone therapy (1 mg/kg, per os, q 12 hr), the dog was afebrile and nonpainful with normal ambulation. Lameness recurred twice over the next 5 mo. At 9 mo of age, diagnostics showed severe erosive polyarthritis of both stifles with an inflammatory leukogram and arthrocentesis findings consistent with septic arthritis, and the dog died despite antibiotic therapy. This is the first case of TNS described in the North American literature, and it is unique in that we had the opportunity to document progression of radiographic abnormalities over more than 6 mo. TNS should be considered in young border collies with signs suggestive of immune-mediated polyarthritis, septic arthritis, or hypertrophic osteodystrophy, combined with neutropenia or gastrointestinal signs.

The HIV Integrase Inhibitor Raltegravir Inhibits Felid Alphaherpesvirus 1 Replication by Targeting both DNA Replication and Late Gene Expression.

Alphaherpesvirus-associated ocular infections in humans caused by human alphaherpesvirus 1 (HHV-1) remain challenging to treat due to the frequency of drug application required and the potential for the selection of drug-resistant viruses. Repurposing on-the-market drugs is a viable strategy to accelerate the pace of drug development. It has been reported that the human immunodeficiency virus (HIV) integrase inhibitor raltegravir inhibits HHV-1 replication by targeting the DNA polymerase accessory factor and limits terminase-mediated genome cleavage of human betaherpesvirus 5 (HHV-5). We have previously shown, both in vitro and in vivo, that raltegravir can also inhibit the replication of felid alphaherpesvirus 1 (FeHV-1), a common ocular pathogen of cats with a pathogenesis similar to that of HHV-1 ocular disease. In contrast to what was reported for HHV-1, we were unable to select for a raltegravir-resistant FeHV-1 strain in order to define any basis for drug action. A candidate-based approach to explore the mode of action of raltegravir against FeHV-1 showed that raltegravir did not impact FeHV-1 terminase function, as described for HHV-5. Instead, raltegravir inhibited DNA replication, similarly to HHV-1, but by targeting the initiation of viral DNA replication rather than elongation. In addition, we found that raltegravir specifically repressed late gene expression independently of DNA replication, and both activities are consistent with inhibition of ICP8. Taken together, these results suggest that raltegravir could be a valuable therapeutic agent against herpesviruses.IMPORTANCE The rise of drug-resistant herpesviruses is a longstanding concern, particularly among immunocompromised patients. Therefore, therapies targeting viral proteins other than the DNA polymerase that may be less likely to lead to drug-resistant viruses are urgently needed. Using FeHV-1, an alphaherpesvirus closely related to HHV-1 that similarly causes ocular herpes in its natural host, we found that the HIV integrase inhibitor raltegravir targets different stages of the virus life cycle beyond DNA replication and that it does so without developing drug resistance under the conditions tested. This shows that the drug could provide a viable strategy for the treatment of herpesvirus infections.

Broad anti-herpesviral activity of α-hydroxytropolones.

Herpesviruses are ubiquitous in animals and cause economic losses concomitant with many diseases. Most of the domestic animal herpesviruses are within the subfamily Alphaherpesvirinae, which includes human herpes simplex virus 1 (HSV-1). Suppression of HSV-1 replication has been reported with α-hydroxytropolones (αHTs), aromatic ring compounds that have broad bioactivity due to potent chelating activity. It is postulated that αHTs inhibit enzymes within the nucleotidyltransferase superfamily (NTS). These enzymes require divalent cations for nucleic acid cleavage activity. Potential targets include the nuclease component of the herpesvirus terminase (pUL15C), a highly conserved NTS-like enzyme that cleaves viral DNA into genomic lengths prior to packaging into capsids. Inhibition of pUL15C activity in biochemical assays by various αHTs previously revealed a spectrum of potencies. Interestingly, the most potent anti-pUL15C αHT inhibited HSV-1 replication to a limited extent in cell culture. The aim of this study was to evaluate three different αHT molecules with varying biochemical anti-pUL15C activity for a capacity to inhibit replication of veterinary herpesviruses (BoHV-1, EHV-1, and FHV-1) and HSV-1. Given the known discordant potencies between anti-pUL15C and HSV-1 replication inhibition, a second objective was to elucidate the mechanism of action of these compounds. The results show that αHTs broadly inhibit herpesviruses, with similar inhibitory effect against HSV-1, BoHV-1, EHV-1, and FHV-1. Based on immunoblotting, Southern blotting, and real-time qPCR, the compounds were found to specifically inhibit viral DNA replication. Thus, αHTs represent a new class of broadly active anti-herpesviral compounds with potential veterinary applications.