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.

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.

Deformity secondary to bilateral incomplete ossification of the humeral condyle in a German Shorthaired Pointer dog. A case report with ten-year follow-up.

A five-month-old male, German Shorthaired Pointer dog was presented for severe, bilateral, thoracic-limb-lameness, with elbow swelling, pain, and crepitus. Radiography and computed tomography confirmed bilateral incomplete ossification of the humeral condyles (IOHC), with a non-displaced incomplete fracture of the left medial epicondylar crest and condylar deformity, characterised by enlargement of the trochleas with extension of the disto-medial aspect of the bone below the normal elbow joint level, and a deformed proximo-medial aspect of the ulna and radius. Transcondylar lag screws were placed bilaterally in an attempt to prevent fracture. Microscopic examination of biopsies, harvested from both humeral condyles, was supportive of IOHC. Six years after surgery, radiographs showed severe osteoarthritis and it appeared that fusion of the humeral condyles had not occurred. The dog followed an active life style until 10 years after surgery when the elbows showed almost no observable range-of-motion. However, discomfort was evident only after heavy exercise. Incomplete ossification of the humeral condyles may be associated with osteoarthritis, risk for non-union, implant failure, or secondary condylar fracture. This dog maintained good limb function despite these concerns, and despite bilateral humeral condylar deformity as well as development of severe osteoarthritis. This is the first report of a German Shorthaired Pointer dog with IOHC and the first histological description supportive of this condition in a five-month-old dog.

Clinical and pathologic features of West Nile virus infection in native North American owls (Family strigidae).

Since the initial report of West Nile virus in the northeastern United States in 1999, the virus has spread rapidly westward and southward across the country. In the summer of 2002, several midwestern states reported increased cases of neurologic disease and mortality associated with West Nile virus infection in various native North American owl species. This report summarizes the clinical and pathologic findings for 13 captive and free-ranging owls. Affected species were all in the family Strigidae and included seven snowy owls (Nyctea scandiaca), four great-horned owls (Bubo virginianus), a barred owl (Strix varia), and a short-eared owl (Asio flammeus). Neurologic signs identified included head tilt, uncoordinated flight, paralysis, tremors, and seizures. Owls that died were screened for flaviviral proteins by immunohistochemical staining of formalin-fixed tissues, followed by specific polymerase chain reaction assay to confirm West Nile virus with fresh tissues when available. Microscopic lesions were widespread, involving brain, heart, liver, kidney, and spleen, and were typically nonsuppurative with infiltration by predominantly lymphocytes and plasma cells. Lesions in owls were much more severe than those previously reported in corvids such as crows, which are considered highly susceptible to infection and are routinely used as sentinel species for monitoring for the presence and spread of West Nile virus. This report is the first detailed description of the pathology of West Nile virus infection in Strigiformes and indicates that this bird family is susceptible to natural infection with West Nile virus.

Effect of experimentally induced type II bovine viral diarrhea virus infection on platelet function in calves.

OBJECTIVE: To evaluate platelet aggregation responses in calves experimentally infected with a thrombocytopenia-inducing type II bovine viral diarrhea virus (BVDV) isolate (BVDV 890). ANIMALS: 9 neonatal male Holstein calves. PROCEDURE: 5 calves were inoculated with BVDV 890, and 4 were used as controls. Platelet aggregation studies and attempts to isolate BVDV from platelets were performed 2 days before, the day of, and every 2 days for 12 days after inoculation. Platelet function was assessed by means of optical aggregometry, using adenosine diphosphate and platelet-activating factor as agonists. Bovine viral diarrhea virus was isolated from purified platelet preparations by use of an immunoperoxidase monolayer assay. RESULTS: Maximum percentage aggregation and slope of the aggregation curve decreased over time in calves infected with BVDV. Bovine viral diarrhea virus was not isolated from platelets from control calves, but it was isolated from infected calves from 4 through 12 days after inoculation. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that platelet function may be depressed in calves infected with type II BVDV. Although the mechanism for altered platelet function was not determined, it likely involved an increase in the percentage of aged platelets in the circulation, a direct virus-platelet interaction, or an indirect virus-platelet interaction. Platelet dysfunction, in addition to thrombocytopenia, may contribute to the hemorrhagic syndrome associated with acute type II BVDV infection in calves.

Disseminated Rhodococcus equi infection in two goats.

Rhodococcus equi infection was diagnosed in two goats from the same herd. At necropsy, numerous caseating granulomas were disseminated throughout the liver, lungs, abdominal lymph nodes, medulla of right humerus, and the right fifth rib of goat No. 1, and the liver of goat No. 2. Histopathologic examination confirmed the presence of multiple caseating granulomas in these organs. Numerous gram-positive and Giemsa-positive coccobacilli were identified within the cytoplasm of macrophages. Aerobic bacterial cultures of the liver and lung from both goats yielded a pure growth of R. equi. R. equi antigens were immunohistochemically identified in caseating granulomas from both goats. However, the 15- to 17-kd virulence antigens of R. equi were not detected, suggesting possible infection by an avirulent strain of this organism.

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 10(7) 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.

Characterization of natural occurring Pneumocystis carinii pneumonia in pigs by histopathology, electron microscopy, in situ hybridization and PCR amplification.

Macroscopic, histologic, ultrastructural, microbiologic, in situ hybridization (ISH) and PCR detection results in three 8-week-old pigs naturally infected with Pneumocystis carinii (PC) are described. All animals had a nonsuppurative interstitial pneumonia and intra-alveolar Pneumocystis organisms with foamy eosinophilic and PAS positive appearance. Ultrastructurally, PC trophozoites and cysts were observed in pigs No. 2 and No. 3, with the former being much more numerous. PC organisms were located on the alveolar surface or within the alveolar septa. Trophozoites had numerous filopodia and were thick-walled. Cysts had no or few filopodia, were thick-walled and contained intracystic bodies. Using non-isotopic ISH on formalin-fixed, paraffin-embedded lung tissue sections, PC DNA from pigs No. 2 and No. 3 hybridized with a probe specific for PC ribosomal RNA (rRNA). Using primers specific for mitochondrial rRNA gene (pAZ102-E/pAZ102-H), and for the internal transcriber spacers of ribosomal gene of PC, PCR methods amplified a product in the lung of pigs No. 2 and No. 3 using either frozen or formalin-fixed and paraffin-embedded lung tissue. DNA from Pig No. 1 samples did not amplify with any primer. This is the first time that molecular biology techniques (in situ hybridization and PCR) have been applied to the study of porcine pneumocystosis.

Surgical treatment of progressive ethmoidal hematoma aided by computed tomography in a foal.

A progressive ethmoidal hematoma (PEH) was treated successfully in a 4-week-old Belgian filly by surgical removal, using a frontonasal bone flap. The filly had respiratory stridor, epistaxis, and facial enlargement over the left paranasal sinuses, which had progressively increased in size since birth. Computed tomographic images of the head obtained with the foal under general anesthesia were useful in determining the extent and nature of the soft-tissue mass and planning surgical intervention. On the basis of the histologic appearance of the mass, a diagnosis of PEH was made. Twelve months after surgery, the facial appearance was normal and the abnormal appearance of the ethmoid region on endoscopic evaluation was less obvious, with return of the nasal septum to a normal position. Progressive ethmoidal hematoma is uncommon and, to our knowledge, has not been reported in a neonate. Clinical signs of PEH in this foal were atypical because of the rapid enlargement of the mass, extent of facial deformity, and minimal epistaxis and interoperative hemorrhage.

A primary platelet disorder of consanguineous simmental cattle.

A severe hereditary hemorrhagic diathesis in Simmental cattle has been identified in North America. Platelet numbers and coagulation profiles of affected cattle are normal. We have further characterized the severe dysfunction of platelet aggregation. All agonists tested elicited normal shape change. Aggregations in response to ADP, A23187, and collagen were absent. Aggregations were decreased or required more time for completion in response to PAF and thrombin. No ultrastructural abnormalities were observed in transmission electron micrographs. Dense granule release of ATP in response to PAF was normal. Thrombin-induced aggregation was dependent upon external calcium concentration in normal but not affected animals. Clot retraction in the blood from affected animals was abnormal. The data implicate a defect of Ca++ mobilization or utilization.

Investigation of an epizootic of bovine viral diarrhea virus infection in calves.

Eight of 19 calves born to bovine viral diarrhea virus (BVDV)-negative and -immunocompetent dams were determined to be infected with a noncytopathic strain of BVDV. Six of the 8 calves had diarrhea and 2 had no clinical signs of disease. In 3 euthanatized calves, lesions consistent with mucosal disease were found throughout the gastrointestinal tract, and the virus was isolated from the spleen, lymph nodes, and small intestine. In 5 calves, BVDV was isolated from mononuclear cells in blood samples obtained 21 days apart, indicating persistent infection. The virus was not isolated from sera obtained from 2 calves, with chronic nonclinical infections, that had neutralizing antibody titers > or = 1:512 against bovine viral diarrhea-Singer virus and titers > or = 1:256 against the persistent BVDV. Twenty-one days after vaccination with a vaccine that contained inactivated noncytopathic and cytopathic biotypes of BVDV, 4 of 5 persistently infected calves had neutralizing antibody titers < or = 1:4 against the bovine viral diarrhea-Singer virus and their persistent virus. Prior to vaccination, 2 of 11 virus-negative calves had neutralizing antibody titers < or = 1:128 against the bovine viral diarrhea-Singer virus, and after vaccination, only 1 virus-negative calf had a titer < or = 1:512. At 149 days after revaccination and prior to weaning, 4 virus-negative calves had neutralizing antibody titers < or = 1:512 (range, 1:16 to 1:384). Under the specific conditions in this herd, we were not able to detect a beneficial effect of vaccination.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

Obstructive enterolith in an 11-month-old miniature horse.

An obstructive enterolith was diagnosed in an 11-month-old 68-kg miniature horse with a 24-hour history of mild, intermittent colic. Abdominal radiography revealed gas distention of the cecum and the right and left ascending colon, and a radiodense elliptical structure in the area of the left kidney. During exploratory laparotomy, the right dorsal colon was noticed to contain ecchymotic hemorrhage and generalized discoloration in the portion proximal to the location of the enterolith. While exteriorizing the ascending colon, it ruptured caudal to the diaphragmatic flexure in the right dorsal colon. Radiography of the enterolith did not reveal a metallic center, but division of the stone allowed identification of a canvas cloth, and chemical analysis determined the stone's composition to be ammonium magnesium phosphate. Although equine enteroliths were reported over 100 years ago and the factors involved in their formation were investigated, the reasons for their development have yet to be determined. The mean reported age of horses requiring abdominal surgery because of an obstructive enterolith is 10 years, with the youngest being 4 years. The time required for an enterolith to enlarge is unknown, although a true enterolith has not been reported in horses younger than 4 years of age.

Efficacy of hydrated sodium calcium aluminosilicate in reducing the toxicity of dietary zearalenone to mink.

Ovariectomized mink were fed diets containing zearalenone (ZEN) at concentrations of 0, 10 or 20 ppm with or without 0.5% hydrated sodium calcium aluminosilicate (HSCAS) for 24 days. Zearalenone at 10 and 20 ppm caused a significant increase in uterine weights, while 20 ppm ZEN resulted in significantly higher vulva swelling scores when compared to controls. The presence of HSCAS in the diet did not alter these hyperestrogenic effects of ZEN. In a second experiment, female mink were provided diets containing 20 ppm ZEN, 20 ppm ZEN plus 0.5% HSCAS or a control diet from 1 January 1989 through whelping (25 April to 15 May 1989). The females were given an opportunity to mate with untreated proven breeder males beginning on 1 March (day 59 of exposure). ZEN did not have an effect on the number of females whelping but there was a significant increase in gestation length, a decrease in litter size and an increase in kit mortality from birth to 3 weeks of age when compared to the control group and the group receiving the combination of ZEN and HSCAS. These results suggest that HSCAS can alleviate some of the reproductive effects of ZEN which are not related to its estrogenic action.

Cerebrospinal nematodiasis in a goat herd.

During the fall of 1985, 4 Angora goats, from a herd of 40, were examined on a farm in central Michigan. Affected goats were alert but had neurologic deficits consistent with upper and lower moto neuron involvement. Eosinophilic pleocytosis in a cerebrospinal fluid sample from one goat was consistent with cerebrospinal nematodiasis. Parelaphostrongylosis was confirmed in 3 goats by identification of Parelaphostrongylosis tenuis larvae in spinal cord sections. Ivermectin may have influenced the herd's susceptibility to new parasitic infections. Control of parelaphostrongylosis probably is best achieved by removal of susceptible animals from treed swamps coinhabited by white-tailed deer from late summer until after the first killing frost.