Correlation between computed tomography and histological evaluation of nodal metastasis in dogs with mast cell tumours.

Early diagnosis of nodal metastasis has been shown to impact prognosis for dogs with mast cell tumours (MCT). The objective of this retrospective study was to determine the correlation between computed tomographic characteristics of lymph nodes and histologic nodal metastasis using the HN classification system, in dogs with cutaneous or subcutaneous MCT and regional lymph node(s) removal. Dogs that had removal of MCT and regional lymphadenectomy within 31 days of the initial staging computed tomography (CT) were enrolled. Subjective lymph node characteristics used included margination, loss of fat at hilus, shape of margin, perinodal fat pattern, increase in number of nodes, and pre- and post-contrast heterogeneity. Enhancement, heterogeneity, and short-long axis ratio were calculated. Seventy-one lymph nodes from 37 dogs were included. Generalised linear mixed model of assessment of lymph node was performed twice, with binary outcome [non-metastatic (HN0/1) versus metastatic (HN2/3)] and 4-point scales (HN0-HN3). After blind assessment of 7 characteristics described above, a final subjective interpretation of each lymph node as non-metastatic or metastatic was assigned. A significant correlation was found between final interpretation and prediction of metastasis. Higher HN classification was also significantly correlated with the increased number of nodes and pre- and post-contrast heterogeneity. No correlation was found in short-long axis ratio, calculated heterogeneity, or degree of enhancement. Sensitivity of CT was 35.7%, specificity was 96.6%, and accuracy was 60.5% for nodal metastasis. CT alone cannot be recommended for assessment of metastasis. The use of multiple computed tomographic characteristics may increase accuracy of nodal metastasis detection.

Associations between urogenital carcinoma and DECA-BDE (BDE-209) among wild California Sea lions (Zalophus californianus) and Steller Sea lions (Eumetopias jubatus).

Urogenital carcinoma (UGC) is prevalent among California sea lions (Zalophus californianus), while less is known concerning UGC among Steller sea lions (Eumetopias jubatus). Our objective was to investigate associations between UGC and polybrominated diphenyl ethers (PBDEs) among both sea lion species. Twenty-nine California sea lions and 20 Steller sea lions were lethally removed from the Columbia River Basin, Oregon, USA between 2020 and 2021, under Section 120 of the Marine Mammal Protection Act. UGC was diagnosed through gross necropsy and histopathology. Forty PBDE congeners were analyzed in blubber, including BDE-209, a potential carcinogen. Twenty (69 %) California sea lions and one (5 %) Steller sea lion were diagnosed with UGC. All cases were identified as early stage UGC, aside from one California sea lion with more advanced stage UGC. Among California sea lions, associations between PBDEs and UGC were analyzed using logistic regression. In the adjusted model, BDE-209 (log2-transformed) was associated with increased odds of UGC [Odds Ratio (OR): 4.68, 95 % confidence interval: 1.04, 21.0, OR p-value = 0.044). This is the first study to report BDE-209 concentrations in sea lion blubber. The percentages of California and Steller sea lions diagnosed with UGC were higher than expected for wild (non-stranded) sea lions. Our results suggested blubber BDE-209 was potentially associated with UGC in California sea lions in the Columbia River Basin.

Trypanosoma cruzi infection diagnosed in dogs in nonendemic areas and results from a survey suggest a need for increased Chagas disease awareness in North America.

OBJECTIVE: To describe the clinical presentation and outcome in dogs diagnosed with Trypanosoma cruzi infection in nonendemic areas and to survey veterinary cardiologists in North America for Chagas disease awareness. ANIMALS: 12 client-owned dogs; 83 respondents from a veterinary cardiology listserv. PROCEDURES: A retrospective, multicenter medical records review to identify dogs diagnosed with American trypanosomiasis between December 2010 and December 2020. An anonymous online survey was conducted August 9 to 22, 2022. RESULTS: Diagnosis was made using indirect fluorescent antibody titer (n = 9), quantitative PCR assay (1), or postmortem histopathology (2). Time spent in Texas was < 1 year (n = 7) or 2 to 8 years (5). Time in nonendemic areas prior to diagnosis was < 1 year (n = 10) and > 3 years (2). Eleven had cardiac abnormalities. Of the 12 dogs, 5 had died unexpectedly (range, 1 to 108 days after diagnosis), 4 were still alive at last follow-up (range, 60 to 369 days after diagnosis), 2 were euthanized because of heart disease (1 and 98 days after diagnosis), and 1 was lost to follow-up. Survey results were obtained from 83 cardiologists in North America, of which the self-reported knowledge about Chagas disease was limited in 49% (41/83) and 69% (57/83) expressed interest in learning resources. CLINICAL RELEVANCE: Results highlight the potential for encountering dogs with T cruzi infection in nonendemic areas and need for raising awareness about Chagas disease in North America.

Comparison of indirect computed tomographic lymphography and near-infrared fluorescence sentinel lymph node mapping for integumentary canine mast cell tumors.

OBJECTIVE: To compare the independent and combined use of indirect computed tomographic lymphography (ICTL) and near-infrared fluorescence (NIRF) for sentinel lymph node (SLN) mapping in dogs with integumentary mast cell tumors (MCT) and report the metastatic LN rate. STUDY DESIGN: Prospective clinical study. ANIMALS: Twenty client-owned dogs. METHODS: Dogs underwent preoperative ICTL, then intraoperative NIRF SLN mapping and excision of the anatomic lymph node (ALN) and/or SLN, and primary MCT. Technique agreement was complete if the same SLN was detected, and partial if the same SLN was detected along with additional SLN. No agreement occurred if the techniques detected different or no SLN. MCT were graded using two- and three-tier schemes, and LN were graded from HN0-3; HN2-3 were considered metastatic. RESULTS: Complete, partial, and no agreement between ICTL and NIRF was seen in 8/20 (40%), 8/20 (40%), and 4/20 (20%) dogs, respectively. Detection of ICTL-SLN and NIRF-SLN failed in 1/20 (5%) and 4/20 (20%), respectively. Tumors were grade II/low-grade in 19/20 (95%) and grade III/high-grade in 1/20 (5%) dogs. Nineteen out of 20 (95%) dogs had HN2-3 LN. CONCLUSIONS: Technique agreement of at least one SLN was seen in 16/20 (80%) dogs. Although most MCT were classified as intermediate to low grade, LN metastases were commonly detected. CLINICAL SIGNIFICANCE: Combining ICTL and NIRF for MCT SLN mapping yields high SLN detection rates. Lymph node metastasis may be more common than previously reported for intermediate to low grade MCT.

Black oil sunflower seed ingestion associated with renal azotemia, gastroesophageal ulceration, and a high mortality rate in four alpacas and two llamas.

CASE DESCRIPTION: 4 alpacas and 2 llamas (11 months to 11 years old) from 2 properties were examined for lethargy (6/6), salivation and regurgitation (4/6), and recumbency (3/6). Signs developed approximately 48 to 72 hours after accidental access to black oil sunflower seeds. CLINICAL FINDINGS: 3 alpacas died suddenly prior to treatment and were necropsied. One llama survived, and 1 alpaca and 1 llama died after days of medical treatment. All 3 treated animals had systemic inflammatory signs including tachycardia, fever, and hematologic changes. Biochemical anomalies included azotemia, hyperglycemia, hyponatremia, hypochloremia, and hypoalbuminemia. Necropsy identified numerous sunflower seeds in the gastrointestinal tract of all 5 animals that died, with pulmonary congestion (5/5 animals), mild centrilobular vacuolar hepatic degeneration (4/5), and erosions of the esophagus (3/5) and first (3/5) and third (1/5) compartments of the forestomach. Renal tubular necrosis was found in the 2 animals that died on day 4 of treatment. TREATMENT AND OUTCOME: One llama responded successfully to intensive medical management including supplemented IV fluid therapy, oral and partial parenteral nutrition, and administration of antimicrobials, furosemide, and insulin and was clinically normal with plasma biochemical analysis values within reference range 12 weeks later. Vitamin D, oxalates, heavy metals, and mycotoxins were excluded as the cause of clinical signs on the basis of screening of uneaten seeds and tissue samples and gastric content from the treated llama that died. CLINICAL RELEVANCE: Inadvertent large volume black oil sunflower seed ingestion resulted in a high mortality rate in camelids. A specific toxic principle was not identified. Feeding this product to camelids is not recommended to avoid the risk of accidental overingestion and subsequent disease.

A case of canine renal lymphoma of granular lymphocytes with severe polycythemia.

BACKGROUND: Renal lymphoma in dogs is rare and has a poor prognosis. Granular lymphocyte morphology is rarely reported in canine renal lymphoma. Mild to moderate polycythemia is reported in a number of canine renal lymphoma cases. CASE PRESENTATION: A 10-year-old Labrador retriever presented to a university veterinary teaching hospital after a 1-month history of polyuria, polydipsia, and pollakiuria and a 2-week history of abdominal distention, lethargy, and increased respiratory effort. Abdominal ultrasound showed a wedge-shaped to rounded, heterogeneously hypoechoic mass lesion in the left kidney. Cytologic analysis of a percutaneous aspirate of the mass was consistent with lymphoma of granular lymphocytes. Severe polycythemia (hematocrit 0.871) was noted on a complete blood cell count. Clonality analysis identified a clonally rearranged T-cell receptor (TCR) gene and immunohistochemical staining was CD3+, CD79a- and CD11d+, supporting cytotoxic T-cell lymphoma. CONCLUSIONS: To our knowledge, this is the first report of renal cytotoxic T-cell lymphoma with severe polycythemia in a dog. Severe polycythemia and renal cytotoxic T-cell lymphoma are both rare in dogs; this report adds to the body of knowledge on these conditions.

Investigating the Effects of Error Management Training versus Error Avoidance Training on the Performance of Veterinary Students Learning Blood Smear Analysis.

Conventional veterinary training emphasizes correct methodologies, potentially failing to exploit learning opportunities that arise as a result of errors. Error management training (EMT) encourages mistakes during low-stakes training, with the intention of modifying perceptions toward errors and using them to improve performance in unfamiliar scenarios (adaptive transfer). Herein, we aimed to determine the efficacy of EMT, supplemented by a metacognitive module, for veterinary students learning blood smear preparation and interpretation. Our hypothesis was that EMT and metacognition are associated with improved adaptive transfer performance, as compared with error avoidance training (EAT). A total of 26 students were prospectively enrolled in this double-blind study. Performance was evaluated according to monolayer area, smear quality, cell identification, calculated white blood cell differential counts, and overall application/interpretation. Students were trained with normal canine blood and static photomicrographs. Participants tested 72 hours after training demonstrated improved performance in a test that directly recapitulated training (Wilcoxon matched-pairs signed-rank test; two-tailed p all ≤ .001). There were no significant differences between EAT and EMT in this test (Mann-Whitney U test and Welch's t-test; two-tailed p ≥ .26) or in short- and long-term adaptive transfer tests (p ≥ .22). Survey data indicate that participants found errors to be a valuable element of training, and that many felt capable of accurately reflecting on their own performance. These data suggest that EMT might produce outcomes comparable to EAT as it relates to blood smear analysis.

Investigating the Effects of Error Management Training versus Error Avoidance Training on the Performance of Veterinary Students Learning to Tie Surgical Knots.

Although errors can be a powerful impetus for learning, conventional pedagogy often emphasizes error-avoidance strategies that reward correct answers and disfavor mistakes. Error management training (EMT) takes an explicitly positive approach to errors, using them to create an active and self-directed learning environment. Using a surgical knot-tying model, we aimed to determine the efficacy of EMT among veterinary students with no prior surgical experience. We hypothesized that EMT would result in improved performance in unfamiliar scenarios (adaptive transfer) compared with an error-avoidance method. In this prospective double-blinded study, 42 students were equally divided between error avoidance training (EAT) and EMT groups. Performance in instrument- and hand-tied knots was evaluated for technique, time, number of attempts, and, when applicable, knot-leaking pressure. All participants demonstrated significant improvement between a pre-test and an analogous test 48 hours after training for all six outcomes (Wilcoxon matched pairs; two-tailed ps ≤ .013). An adaptive transfer test found no significant differences between EMT and EAT at 48 hours (ps ≥ .053). All participants demonstrated a significant performance decline in six of eight outcomes at 7 weeks post-training (ps ≤ .021). This decline was not significant for four of six EMT outcomes yet significant for five of six EAT outcomes. These data suggest that students trained in both EMT and EAT experience comparable gains in short-term performance, including adaptive transfer. Compared with EAT, EMT may help attenuate performance decline after a sustained period of quiescence. Educators may consider actively incorporating EMT into veterinary curricula.

Comparative Pathology of the Peripheral Nervous System.

The peripheral nervous system (PNS) relays messages between the central nervous system (brain and spinal cord) and the body. Despite this critical role and widespread distribution, the PNS is often overlooked when investigating disease in diagnostic and experimental pathology. This review highlights key features of neuroanatomy and physiology of the somatic and autonomic PNS, and appropriate PNS sampling and processing techniques. The review considers major classes of PNS lesions including neuronopathy, axonopathy, and myelinopathy, and major categories of PNS disease including toxic, metabolic, and paraneoplastic neuropathies; infectious and inflammatory diseases; and neoplasms. This review describes a broad range of common PNS lesions and their diagnostic criteria and provides many useful references for pathologists who perform PNS evaluations as a regular or occasional task in their comparative pathology practice.

A COL7A1 Variant in a Litter of Neonatal Basset Hounds with Dystrophic Epidermolysis Bullosa.

We investigated three neonatal Basset Hound littermates with lesions consistent with epidermolysis bullosa (EB), a group of genetic blistering diseases. A clinically normal bitch was bred to her grandfather by artificial insemination. Out of a litter of seven puppies, two affected puppies died and one was euthanized, with these puppies being submitted for diagnostic necropsy. All had multiple bullae and ulcers involving the nasal planum and paw pads, as well as sloughing claws; one puppy also had oral and esophageal ulcers. The complete genome of one affected puppy was sequenced, and 37 known EB candidate genes were assessed. We found a candidate causative variant in COL7A1, which encodes the collagen VII alpha 1 chain. The variant is a complex rearrangement involving duplication of a 107 bp region harboring a frameshift deletion of 7 bp. The variant is predicted to truncate more than 75% of the open reading frame, p.(Val677Serfs*11). Targeted genotyping of this duplication confirmed that all three affected puppies were homozygous for the duplication, whereas 12 unaffected Basset Hounds did not carry the duplication. This variant was also not seen in the genomes of more than 600 dogs of other breeds. COL7A1 variants have been identified in humans and dogs with dystrophic epidermolysis bullosa (DEB). The identified COL7A1 variant therefore most likely represents the causative variant and allows the refinement of the preliminary EB diagnosis to DEB.

In Vivo Aortic Magnetic Resonance Elastography in Abdominal Aortic Aneurysm: A Validation in an Animal Model.

OBJECTIVES: Using maximum diameter of an abdominal aortic aneurysm (AAA) alone for management can lead to delayed interventions or unnecessary urgent repairs. Abdominal aortic aneurysm stiffness plays an important role in its expansion and rupture. In vivo aortic magnetic resonance elastography (MRE) was developed to spatially measure AAA stiffness in previous pilot studies and has not been thoroughly validated and evaluated for its potential clinical value. This study aims to evaluate noninvasive in vivo aortic MRE-derived stiffness in an AAA porcine model and investigate the relationships between MRE-derived AAA stiffness and (1) histopathology, (2) uniaxial tensile test, and (3) burst testing for assessing MRE's potential in evaluating AAA rupture risk. MATERIALS AND METHODS: Abdominal aortic aneurysm was induced in 31 Yorkshire pigs (n = 226 stiffness measurements). Animals were randomly divided into 3 cohorts: 2-week, 4-week, and 4-week-burst. Aortic MRE was sequentially performed. Histopathologic analyses were performed to quantify elastin, collagen, and mineral densities. Uniaxial tensile test and burst testing were conducted to measure peak stress and burst pressure for assessing the ultimate wall strength. RESULTS: Magnetic resonance elastography-derived AAA stiffness was significantly higher than the normal aorta. Significant reduction in elastin and collagen densities as well as increased mineralization was observed in AAAs. Uniaxial tensile test and burst testing revealed reduced ultimate wall strength. Magnetic resonance elastography-derived aortic stiffness correlated to elastin density (ρ = -0.68; P < 0.0001; n = 60) and mineralization (ρ = 0.59; P < 0.0001; n = 60). Inverse correlations were observed between aortic stiffness and peak stress (ρ = -0.32; P = 0.0495; n = 38) as well as burst pressure (ρ = -0.55; P = 0.0116; n = 20). CONCLUSIONS: Noninvasive in vivo aortic MRE successfully detected aortic wall stiffening, confirming the extracellular matrix remodeling observed in the histopathologic analyses. These mural changes diminished wall strength. Inverse correlation between MRE-derived aortic stiffness and aortic wall strength suggests that MRE-derived stiffness can be a potential biomarker for clinically assessing AAA wall status and rupture potential.

Nasal adenocarcinoma as a suspected secondary malignant neoplasm in a cat previously treated for nasal lymphoma.

CASE SUMMARY: A case of nasal adenocarcinoma as a suspected secondary malignant neoplasm following definitive radiation therapy and multiagent chemotherapy for nasal lymphoma is described. An 11-year-old spayed female domestic shorthair cat was presented for a 3-week history of progressive facial swelling located over the nasal planum and extending to the medial canthus of the right eye. The cat was previously diagnosed with nasal lymphoma and treated with chemotherapy and definitive radiation 2.5 years prior. Although a definitive diagnosis could not be obtained via cytology, recurrent lymphoma was suspected based on the cat's history and recurrent clinical signs. A lymphoma-directed chemotherapy protocol was attempted, but no clinical response was achieved. The cat was euthanased owing to progressive clinical signs and a diagnosis of nasal adenocarcinoma was made on necropsy examination. Both the original diagnosis of nasal lymphoma and the secondary diagnosis of nasal adenocarcinoma were confirmed with immunohistochemistry. RELEVANCE AND NOVEL INFORMATION: Secondary malignant neoplasm following radiation therapy is infrequently reported in the veterinary literature. In the few reports that exist, most have described sarcoma development in the dog following radiation therapy. In the present report, we describe a cat with a suspected radiation-induced nasal adenocarcinoma that developed 2.5 years after definitive radiation treatment for nasal lymphoma.

Long-term outcomes of dogs undergoing surgical resection of mast cell tumors and soft tissue sarcomas: A prospective 2-year-long study.

OBJECTIVE: Report clinical outcomes of dogs with surgically excised mast cell tumors (MCT) and soft tissue sarcomas (STS). STUDY DESIGN: Prospective clinical study. SAMPLE POPULATION: Fifty-three dogs with 52 MCT (50 low grade, 2 high grade) and 19 STS (12 grade I, 6 grade II, 1 grade III). METHODS: All dogs were examined at 3, 6, 12, 18, and 24 months postoperatively, with cytologic or histopathologic evaluation of suspected local recurrences. Dogs euthanized because of study tumor-related causes underwent necropsy. RESULTS: Median intraoperative margins were 20 mm and 30 mm wide for MCT and STS, respectively, with 1 fascial plane resected en bloc. The narrowest histologic tumor-free margins measured <1 mm in 21 of 52 (40%) MCT and 7 of 19 (37%) STS. All dogs were followed for 24 months. Two of 50 (4%) low-grade MCT were diagnosed, with local recurrence 181 and 265 days postoperatively. Two of 36 (6%) dogs with low-grade MCT developed visceral metastasis 181 and 730 days postoperatively. One of 2 dogs with high-grade MCT developed local recurrence 115 days postoperatively. No local recurrence or metastasis was diagnosed after excision of 19 STS. CONCLUSION: Local recurrence rates among predominantly low- to intermediate-grade MCT and STS were low, despite a high prevalence of histologic tumor-free margins <1 mm. Surgical recommendations for high-grade tumors cannot be extrapolated from this population. CLINICAL SIGNIFICANCE: Surgeons should seek to achieve microscopically complete excision for MCT and STS while minimizing patient morbidity and considering limitations of histopathology in predicting outcomes.

Sensitivity of Serologic Testing for Dogs Diagnosed with Coccidioidomycosis on Histology: 52 Cases (2012-2013).

Coccidioidomycosis, or Valley Fever, is a fungal infection caused by Coccidioides spp., soil-living fungi endemic to the southwest region of the United States. The infection can cause various diseases including respiratory, neurologic, cardiac, dermatologic, and ocular disease as well as osteomyelitis in dogs and many other mammals. Obtaining a definitive diagnosis can be challenging. Serology is commonly used as a screening diagnostic test for disease, but both false-negative and false-positive results have been reported. Fifty-two cases of coccidioidomycosis diagnosed via histopathology were retrospectively evaluated. The sensitivity of serology in the study population was determined to be 87% for immunoglobulin G and 46% for immunoglobulin M. The cases were evaluated for an association between negative serology results and anatomic location of disease, but these variables were found to be independent. This study reports the sensitivity of serology for canine coccidioidomycosis and highlights the importance of using multiple diagnostic tests for definitive diagnosis of infection.

A case of canine high-grade T-cell lymphoma immunophenotypically consistent with T-zone lymphoma.

A 9-year-old spayed female Curly Coated Retriever was referred for evaluation of generalized peripheral lymphadenomegaly. The dog was clinically healthy on presentation with no anomalies detected on complete blood count, serum biochemistry, urinalysis, or three-view thoracic radiographs. Fine-needle aspiration (FNA) and cytology of the peripheral lymph nodes were consistent with lymphoma with an intermediate-sized lymphoid population. Flow cytometry of peripheral lymph nodes was consistent with a homogeneous population of CD4+ T cells that had lost expression of the pan-leukocyte antigen CD45. Variable expression of CD21, CD25, and class II major histocompatibility complex (MHC) were also noted. This was considered consistent with T-zone lymphoma (TZL), although the T cells were noted to be larger than usual based on flow cytometry. Due to the suspected indolent nature of this patient's disease and clinical progression, a careful monitoring approach was initially discussed with the owner. However, additional diagnostic testing was performed to confirm the diagnosis. Bone marrow cytology did not show any significant anomalies. The largest lymph node (left mandibular) was extirpated and submitted for histopathology. Based on the lymph node architecture, cellular features, and high mitotic activity, an unexpected diagnosis of peripheral T-cell lymphoma not otherwise specified (PTCL-NOS) was made. The dog was started on CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) chemotherapy. This case illustrates the limitations of using flow cytometry as the sole means of diagnosing TZL and highlights the importance of using complementary tests when subtyping canine lymphoma, which is significant when considering a patient's treatment plan and prognosis.

Comparing the Tolerability of a Novel Wound Closure Device Using a Porcine Wound Model.

Objective: To compare the tolerability and mechanical tensile strength of acute skin wounds closed with nylon suture plus a novel suture bridge device (SBD) with acute skin wounds closed with nylon suture in a porcine model. Approach: Four Yucatan pigs each received 12 4.5 cm full-thickness incisions that were closed with 1 of 4 options: Suture bridge with nylon, suture bridge with nylon and subdermal polyglactin, nylon simple interrupted, and nylon simple interrupted with subdermal polyglactin. Epithelial reaction, inflammation, and scarring were examined histologically at days 10 and 42. Wound strength was examined mechanically at days 10 and 42 on ex vivo wounds from euthanized pigs. Results: Histopathology in the suture entry/exit planes showed greater dermal inflammation with a simple interrupted nylon suture retained for 42 days compared with the SBD retained for 42 days (p < 0.03). While tensile wound strength in the device and suture groups were similar at day 10, wounds closed with the devices were nearly 8 times stronger at day 42 compared with day 10 (p < 0.001). Innovation: A novel SBD optimized for cutaneous wound closure that protects the skin surface from suture strands, forms a protective bridge over the healing wound edges, and knotlessly clamps sutures. Conclusion: This study suggests that the use of a SBD increases the tolerability of nylon sutures in porcine acute skin wound closures allowing for prolonged mechanical support of the wound. For slow healing wounds, this may prevent skin wound disruption, such as edge necrosis and dehiscence.

Immunohistochemical detection of p53, PTEN, Rb, and p16 in canine osteosarcoma using tissue microarray.

Although inactivating mutations of tumor suppressor genes are well described in cell lines of canine osteosarcoma (OS), expression of tumor suppressor proteins in spontaneous disease is poorly characterized. We determined the immunohistochemical expression of p53, PTEN, Rb, and p16 in a large cohort of dogs with OS. Formalin-fixed, paraffin-embedded samples of canine OS were analyzed retrospectively. Primary tumor samples from 145 dogs, collected between 2003 and 2008, were evaluated by tissue microarray. Streptavidin-biotin complex immunohistochemistry was performed using monoclonal antibodies for Rb and PTEN and polyclonal antibodies for p16 and p53. The average age of dogs was 7.6 y, and 118 of 145 (81%) were purebred. Most commonly represented purebreds were Greyhound (23%), Rottweiler (11%), and Labrador Retriever (10%). Immunohistochemical detection of p53, PTEN, Rb, and p16 was 81%, 61%, 66%, and 66%, respectively. The staining pattern for p16 was primarily cytoplasmic; the predominant pattern for PTEN, Rb, and p53 was cytoplasmic and nuclear. Exclusively cytoplasmic staining was noted in 19% of samples positive for p53 and 8% of samples positive for Rb. Kaplan-Meier curves showed that protein expression was not associated with significant differences in overall survival ( p > 0.191). We documented heterogeneity in both immunostaining and subcellular localization of tumor suppressor proteins, providing further characterization of canine OS.

Histologic processing artifacts and inter-pathologist variation in measurement of inked margins of canine mast cell tumors.

Although quantitative assessment of margins is recommended for describing excision of cutaneous malignancies, there is poor understanding of limitations associated with this technique. We described and quantified histologic artifacts in inked margins and determined the association between artifacts and variance in histologic tumor-free margin (HTFM) measurements based on a novel grading scheme applied to 50 sections of normal canine skin and 56 radial margins taken from 15 different canine mast cell tumors (MCTs). Three broad categories of artifact were 1) tissue deformation at inked edges, 2) ink-associated artifacts, and 3) sectioning-associated artifacts. The most common artifacts in MCT margins were ink-associated artifacts, specifically ink absent from an edge (mean prevalence: 50%) and inappropriate ink coloring (mean: 45%). The prevalence of other artifacts in MCT skin was 4-50%. In MCT margins, frequency-adjusted kappa statistics found fair or better inter-rater reliability for 9 of 10 artifacts; intra-rater reliability was moderate or better in 9 of 10 artifacts. Digital HTFM measurements by 5 blinded pathologists had a median standard deviation (SD) of 1.9 mm (interquartile range: 0.8-3.6 mm; range: 0-6.2 mm). Intraclass correlation coefficients demonstrated good inter-pathologist reliability in HTFM measurement (κ = 0.81). Spearman rank correlation coefficients found negligible correlation between artifacts and HTFM SDs ( r ≤ 0.3). These data confirm that although histologic artifacts commonly occur in inked margin specimens, artifacts are not meaningfully associated with variation in HTFM measurements. Investigators can use the grading scheme presented herein to identify artifacts associated with tissue processing.

Reductions in margin length after excision of grade II mast cell tumors and grade I and II soft tissue sarcomas in dogs.

OBJECTIVE: Quantify changes in the circumferential lengths of surgical margins of resected canine mast cell tumors (MCT) and soft tissue sarcomas (STS) between the time of collection and histopathology. STUDY DESIGN: Prospective, hypothesis-driven, clinical study. SAMPLE POPULATION: Two hundred and thirty-seven margins from 69 excised tumors (50 MCT and 19 STS) in 51 client-owned dogs. METHODS: The lengths of surgical margins were recorded (eg, cranial, caudal, dorsal, and ventral) for each tumor at 5 time points: intraoperatively (in vivo), immediately after excision (ex vivo), after formalin fixation (postfixation), once mounted on glass slides (subgross), and as histologically tumor-free margins (HTFMs). RESULTS: Compared to in vivo dimensions, the length of surgical margins at each processing step (ie, ex vivo, postfixation, subgross, and HTFM) was reduced by a median of 3.0, 5.0, 6.0, and 8.8 mm for MCT; 2.5, 2.0, 5.0, and 5.0 mm for STS. All processing steps resulted in significant reductions among MCT samples (P < .0001), except between postfixation vs subgross, and for STS samples (P < .0001), except between ex vivo vs postfixation and subgross vs HTFM. The maximum reduction in the total length of margins (from in vivo to HTFM) was 29.6 and 24.2 mm for MCT and STS, respectively. CONCLUSION: Surgical margin length reductions occur due to a combination of physical factors (eg, tissue elasticity, myofibril contraction, and histologic processing) and biological factors (eg, microscopic tumor infiltration into the grossly normal surgical margin). CLINICAL SIGNIFICANCE: These data provide information relevant to evidence-based surgical planning and may influence patient morbidity in the most commonly encountered cutaneous malignancies of dogs.

Analysis of stress factors associated with KHV reactivation and pathological effects from KHV reactivation.

Koi herpesvirus (KHV) is a highly pathogenic virus of common carp and koi. KHV becomes latent in recovered koi or exposed koi without symptoms, and the latent infection can reactivate under stress conditions. KHV reactivation from latency often occurs when water temperature rapidly rises above 17°C. Dissolved O2 is lower at ≥17°C than at non-stress temperatures ≤15°C. To determine whether reduced dissolved O2 level has a role in KHV reactivation during temperature stress, KHV reactivation was investigated in KHV latently infected koi (KHV+ koi) under stress temperatures by maintaining dissolved O2 consistent with the O2 level at 15°C. There was no significant difference in the amount of reactivated virus between KHV+ koi maintained with and without O2 supplementation during temperature stress. Both handling and sampling were found to be stressful to koi and can contribute to KHV reactivation from latency. There was an increase in KHV genome within white blood cells (WBC) during KHV reactivation, which is about 3-4 fold higher than the amount of KHV genome detectable in WBC during the latency stage. At day 15 post-temperature stress (PTS), inflammation and necrosis were observed in multiple tissues, especially in the gills, eye, intestine, skin and kidney. KHV DNA was also detectable in multiple tissues on days 6, 9 and 15 PTS. Following day 3 PTS, the plasma cortisol levels were higher than that observed in koi before temperature stress, suggesting that KHV reactivation is associated with physiological stress in KHV+ koi.