Barbara A. Steficek

Proposal of a 2-Tier Histologic Grading System for Canine Cutaneous Mast Cell Tumors to More Accurately Predict Biological Behavior

Currently, prognostic and therapeutic determinations for canine cutaneous mast cell tumors (MCTs) are primarily based on histologic grade. However, the use of different grading systems by veterinary pathologists and institutional modifications make the prognostic value of histologic grading highly questionable. To evaluate the consistency of microscopic grading among veterinary pathologists and the prognostic significance of the Patnaik grading system, 95 cutaneous MCTs from 95 dogs were graded in a blinded study by 28 veterinary pathologists from 16 institutions. Concordance among veterinary pathologists was 75% for the diagnosis of grade 3 MCTs and less than 64% for the diagnosis of grade 1 and 2 MCTs. To improve concordance among pathologists and to provide better prognostic significance, a 2-tier histologic grading system was devised. The diagnosis of high-grade MCTs is based on the presence of any one of the following criteria: at least 7 mitotic figures in 10 high-power fields (hpf); at least 3 multinucleated (3 or more nuclei) cells in 10 hpf; at least 3 bizarre nuclei in 10 hpf; karyomegaly (ie, nuclear diameters of at least 10% of neoplastic cells vary by at least two-fold). Fields with the highest mitotic activity or with the highest degree of anisokaryosis were selected to assess the different parameters. According to the novel grading system, high-grade MCTs were significantly associated with shorter time to metastasis or new tumor development, and with shorter survival time. The median survival time was less than 4 months for high-grade MCTs but more than 2 years for low-grade MCTs.

Effects of negative pressure wound therapy on healing of open wounds in dogs.

OBJECTIVE To compare the effect of negative pressure wound therapy (NPWT) with standard-of-care management on healing of acute open wounds in dogs. STUDY DESIGN Prospective, controlled, experimental study. ANIMALS Adult dogs (n=10). METHODS Full-thickness 4 m × 2 m wounds were surgically created on each antebrachium and in each dog were randomized to receive either NPWT or standard wound dressings (CON) for 21 days. Dressing changes and wound evaluations were made at 8 time points. First appearance of granulation tissue, smoothness of granulation tissue, exuberance, percent epithelialization, and percent contraction were compared. Biopsies for histopathology were taken, and histologic scores determined, at 5 time points, and aerobic bacterial wound cultures performed at 2 time points. RESULTS Granulation tissue appeared significantly earlier, and was smoother and less exuberant in NPWT wounds compared with CON wounds. Percent contraction in NPWT wounds was less than CON wounds after Day 7. Percent epithelialization in NPWT wounds was less than CON wounds on Days 11, 16, 18, and 21. Histologic scores for acute inflammation were higher in NPWT on Day 3, and lower on Day 7, than CON wounds. Bacterial load was higher in NPWT on Day 7. CONCLUSION NPWT accelerated appearance of smooth, nonexuberant granulation tissue; however, prolonged use of NPWT impaired wound contraction and epithelialization.

Recommended Guidelines for Submission, Trimming, Margin Evaluation, and Reporting of Tumor Biopsy Specimens in Veterinary Surgical Pathology

Neoplastic diseases are typically diagnosed by biopsy and histopathological evaluation. The pathology report is key in determining prognosis, therapeutic decisions, and overall case management and therefore requires diagnostic accuracy, completeness, and clarity. Successful management relies on collaboration between clinical veterinarians, oncologists, and pathologists. To date there has been no standardized approach or guideline for the submission, trimming, margin evaluation, or reporting of neoplastic biopsy specimens in veterinary medicine. To address this issue, a committee consisting of veterinary pathologists and oncologists was established under the auspices of the American College of Veterinary Pathologists Oncology Committee. These consensus guidelines were subsequently reviewed and endorsed by a large international group of veterinary pathologists. These recommended guidelines are not mandated but rather exist to help clinicians and veterinary pathologists optimally handle neoplastic biopsy samples. Many of these guidelines represent the collective experience of the committee members and consensus group when assessing neoplastic lesions from veterinary patients but have not met the rigors of definitive scientific study and investigation. These questions of technique, analysis, and evaluation should be put through formal scrutiny in rigorous clinical studies in the near future so that more definitive guidelines can be derived.

The Effect of Negative Pressure Wound Therapy on the Healing of Open Wounds in Dogs

OBJECTIVE To compare the effect of negative pressure wound therapy (NPWT) with standard-of-care management on healing of acute open wounds in dogs. STUDY DESIGN Prospective, controlled, experimental study. ANIMALS Adult dogs (n=10). METHODS Full-thickness 4 m × 2 m wounds were surgically created on each antebrachium and in each dog were randomized to receive either NPWT or standard wound dressings (CON) for 21 days. Dressing changes and wound evaluations were made at 8 time points. First appearance of granulation tissue, smoothness of granulation tissue, exuberance, percent epithelialization, and percent contraction were compared. Biopsies for histopathology were taken, and histologic scores determined, at 5 time points, and aerobic bacterial wound cultures performed at 2 time points. RESULTS Granulation tissue appeared significantly earlier, and was smoother and less exuberant in NPWT wounds compared with CON wounds. Percent contraction in NPWT wounds was less than CON wounds after Day 7. Percent epithelialization in NPWT wounds was less than CON wounds on Days 11, 16, 18, and 21. Histologic scores for acute inflammation were higher in NPWT on Day 3, and lower on Day 7, than CON wounds. Bacterial load was higher in NPWT on Day 7. CONCLUSION NPWT accelerated appearance of smooth, nonexuberant granulation tissue; however, prolonged use of NPWT impaired wound contraction and epithelialization.

Effects of negative pressure wound therapy on healing of free full-thickness skin grafts in dogs.

OBJECTIVE To compare healing of free, full-thickness, meshed skin grafts under negative pressure wound therapy (NPWT) with bolster dressings in dogs. STUDY DESIGN Randomized, controlled experimental study, paired design. ANIMALS Dogs (n = 5) METHODS: Full-thickness skin wounds (4 cm × 1.5 cm) were created bilaterally on the antebrachia of 5 dogs (n = 10). Excised skin was grafted to the contralateral limb. Grafts were randomized to NPWT or bolster dressings (control; CON). NPWT was applied continuously for 7 days. Grafts were evaluated on Days 2, 4, 7, 10, 14, and 17, biopsied on days 0, 4, 7, and 14, and had microbial culture on Day 7. Outcome variables were: time to first appearance of granulation tissue, percent graft necrosis, and percent open mesh. Significance was set at P < .05. Histologic findings, culture results, and graft appearance were reported. RESULTS Granulation tissue appeared earlier in the NPWT grafts compared with CON grafts. Percent graft necrosis and remaining open mesh area were both greater in CON grafts compared with NPWT grafts at most time points. Histologic results showed no significant difference in all variables measured, and all cultures were negative. CONCLUSIONS Variables of graft acceptance were superior when NPWT was used in the first week post-grafting. Fibroplasia was enhanced, open meshes closed more rapidly and less graft necrosis occurred with NPWT application. More preclinical studies are required to evaluate histologic differences.

Effect of porcine small intestinal submucosa on acute full-thickness wounds in dogs.

OBJECTIVE To evaluate the effects of porcine small intestinal submucosa (PSIS) on the healing of full-thickness wounds in dogs, specifically the appearance of granulation tissue, percent epithelialization and contraction, histologic variables of inflammation and repair, and aerobic culture results. STUDY DESIGN Prospective, controlled, experimental study. ANIMALS Purpose-bred, female dogs (n=10). METHODS Wounds were created bilaterally on the trunk; 1 side as a control and 1 treated with PSIS. First appearance of granulation tissue was recorded. Total wound area, open wound area, and epithelialized area were measured at 21 time points-wound contraction and percent epithelialization were calculated. Aerobic cultures were taken at 4 time points and wound biopsies at 8. Histologic features were graded into an Acute Inflammation Score and Repair Score. RESULTS There was no difference in first appearance of granulation tissue between PSIS-treated and control wounds. Wound contraction was significantly faster in control wounds as was percent epithelialization after day 21. Histologic Acute Inflammation Scores were significantly higher in PSIS-treated wounds compared with control wounds on days 2 and 6. There were no differences in Histologic Repair Scores between PSIS-treated and control wounds or in aerobic culture results. CONCLUSION Wounds treated with PSIS contract more slowly, epithelialize less, and have more pronounced acute inflammation after implantation than control wounds. CLINICAL RELEVANCE Acute, full-thickness wounds in dogs do not benefit from treatment with PSIS.

Experimental model of type II bovine viral diarrhea virus- induced thrombocytopenia in neonatal calves

Thrombocytopenia has been associated with type II bovine viral diarrhea virus (BVDV) infection in immunocompetent cattle, but the mechanism is unknown. The purpose of the present study was to develop and characterize a model of type II BVDV-induced thrombocytopenia. Colostrum-deprived Holstein calves were obtained immediately after birth, given a BVDV-negative and BVDV antibody-negative plasma transfusion, housed in an isolation facility, and randomly assigned to either control (n = 4) or infected (n = 5) groups. Infected calves were inoculated by intranasal instillation on day 3 of age with 107 TCID50 of the prototype type II isolate, BVDV 890, whereas control calves were sham inoculated. Blood counts and virus isolations from serum, white blood cells, and platelets were performed daily until day 12 after infection, at which time all experimental calves were euthanatized, and pathologic, virologic, and immunohistochemical examinations were performed. On physical examination, the control calves remained normal, but the infected calves developed pyrexia and diarrhea characteristic of type II BVDV infection. The platelet count decreased in all infected calves, and a statistically significant difference in the platelet count between control and infected calves was observed on days 7–12 after infection. In addition, the mean platelet volume and white blood cell counts also decreased. Examination of the bone marrow from the infected calves revealed immunohistochemical staining for BVDV antigen in megakaryocytes and evidence of concurrent megakaryocyte necrosis and hyperplasia.

A 12-year retrospective study of equine abortion in Michigan.

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Hemorrhagic Diathesis Associated with a Hereditary Platelet Disorder in Simmental Cattle

A severe bleeding disorder in Simmental cattle has been described in widespread locations in the USA and Canada. The clinical findings are consistent with a hemophilia-like disease or, more precisely, a hereditary hemorrhagic diathesis and include spontaneous epistaxis, hematuria, and excessive bleeding associated with trauma or standard management procedures such as tattooing, ear tagging, and castration. A preliminary investigation of this defect showed that blood-platelet numbers and coagulation profiles of affected cattle were normal. Affected animals have a marked dysfunction of platelets (thrombopathy), termed Simmental hereditary thrombopathy. The defect is very similar or identical to that described in the same breed by 2 other laboratories.

Ocular melanosis in the Cairn Terrier: histopathological description of the condition, and immunohistological and ultrastructural characterization of the characteristic pigment-laden cells

OBJECTIVE To describe the microscopic features and lineage of proliferating/infiltrating pigmented cells in ocular melanosis of Cairn Terriers. Animals studied Forty-nine globes removed from 45 Cairn Terriers with ocular melanosis and three globes from control dogs were available for microscopic examination. PROCEDURES All globes were examined histologically, eight affected and three control globes were also examined by immunohistochemistry, and three affected and three control globes by transmission electron microscopy. RESULTS Large round pigment-laden cells infiltrated the anterior uvea, obscured the drainage angle and were present within the sclera and episclera of affected globes. Similar pigmented cells were present in lower numbers in the posterior segment of the globe, the optic nerve meninges and periphery of the optic nerve. Changes due to chronic glaucoma were present in many globes and some had evidence of uveitis. Many of the pigmented cells were immunoreactive to HMB45 and some were MITF and vimentin positive. One globe, which was inflamed when removed, had many pigmented cells that were CD18 immunoreactive. The other eyes had lower numbers of CD18 positive cells. The pigmented cells were not immunoreactive to smooth muscle actin, S-100, MART/Melan A, chromogranin A/B, PGP 9.5, synaptophysin, MNF116, AE1/AE3, and CD45. Ultrastructurally many of the pigmented cells had features typical of melanocytes while a smaller number appeared to be melanophages. CONCLUSIONS Ocular melanosis in Cairn Terriers is characterized by an infiltration of pigment-laden cells predominantly, but not exclusively, within the anterior uvea and anterior sclera. Most of these cells appear to be melanocytes although a variable proportion are pigment-laden melanophages.