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Counting mitotic figures (MF) in hematoxylin and eosin–stained histologic sections is an integral part of the diagnostic pathologist’s tumor evaluation. The mitotic count (MC) is used alone or as part of a grading scheme for assessment of prognosis and clinical decisions. Determining MCs is subjective, somewhat laborious, and has interobserver variation. Proposals for standardizing this parameter in the veterinary field are limited to terminology (use of the term MC) and area (MC is counted in an area measuring 2.37 mm2). Digital imaging techniques are now commonplace and widely used among veterinary pathologists, and field of view area can be easily calculated with digital imaging software. In addition to standardizing the methods of counting MF, the morphologic characteristics of MF and distinguishing atypical mitotic figures (AMF) versus mitotic-like figures (MLF) need to be defined. This article provides morphologic criteria for MF identification and for distinguishing normal phases of MF from AMF and MLF. Pertinent features of digital microscopy and application of computational pathology (CPATH) methods are discussed. Correct identification of MF will improve MC consistency, reproducibility, and accuracy obtained from manual (glass slide or whole-slide imaging) and CPATH approaches.

The mitotic count (MC) is an important histological parameter for prognostication of malignant neoplasms. However, it has inter- and intra-observer discrepancies due to difficulties in selecting the region of interest (MC-ROI) and in identifying/classifying mitotic figures (MFs). Recent progress in the field of artificial intelligence has allowed the development of high-performance algorithms that may improve standardization of the MC. As algorithmic predictions are not flawless, the computer-assisted review by pathologists may ensure reliability. In the present study we have compared partial (MC-ROI preselection) and full (additional visualization of MF candidate proposal and display of algorithmic confidence values) computer-assisted MC analysis to the routine (unaided) MC analysis by 23 pathologists for whole slide images of 50 canine cutaneous mast cell tumors (ccMCTs). Algorithmic predictions aimed to assist pathologists in detecting mitotic hotspot locations, reducing omission of MF and improving classification against imposters. The inter-observer consistency for the MC significantly increased with computer assistance (interobserver correlation coefficient, ICC = 0.92) compared to the unaided approach (ICC = 0.70). Classification into prognostic stratifications had a higher accuracy with computer assistance. The algorithmically preselected MC-ROIs had a consistently higher MCs than the manually selected MC-ROIs. Compared to a ground truth (developed with immunohistochemistry for phosphohistone H3), pathologist performance in detecting individual MF was augmented when using computer assistance (F1-score of 0.68 increased to 0.79) with a reduction in false negatives by 38%. The results of this study prove that computer assistance may lead to a more reproducible and accurate MCs in ccMCTs.

Standardization of tumor assessment lays the foundation for validation of grading systems, permits reproducibility of oncologic studies among investigators, and increases confidence in the significance of study results. Currently, there is minimal methodological standardization for assessing tumors in veterinary medicine, with few attempts to validate published protocols and grading schemes. The current article attempts to address these shortcomings by providing standard guidelines for tumor assessment parameters and protocols for evaluating specific tumor types. More detailed information is available in the Supplemental Files, the intention of which is 2-fold: publication as part of this commentary, but more importantly, these will be available as “living documents” on a website ( www.vetcancerprotocols.org ), which will be updated as new information is presented in the peer-reviewed literature. Our hope is that veterinary pathologists will agree that this initiative is needed, and will contribute to and utilize this information for routine diagnostic work and oncologic studies. Journal editors and reviewers can utilize checklists to ensure publications include sufficient detail and standardized methods of tumor assessment. To maintain the relevance of the guidelines and protocols, it is critical that the information is periodically updated and revised as new studies are published and validated with the intent of providing a repository of this information. Our hope is that this initiative (a continuation of efforts published in this journal in 2011) will facilitate collaboration and reproducibility between pathologists and institutions, increase case numbers, and strengthen clinical research findings, thus ensuring continued progress in veterinary oncologic pathology and improving patient care.

Melanoma represents a significant malignancy in humans and dogs. Different from genetically engineered models, sporadic canine melanocytic neoplasms share several characteristics with human disease that could make dogs a more relevant preclinical model. Canine melanomas rarely arise in sun-exposed sites. Most occur in the oral cavity, with a subset having intra-epithelial malignant melanocytes mimicking the in situ component of human mucosal melanoma. The spectrum of canine melanocytic neoplasia includes benign lesions with some analogy to nevi, as well as invasive primary melanoma, and widespread metastasis. Growing evidence of distinct subtypes in humans, differing in somatic and predisposing germ-line genetic alterations, cell of origin, epidemiology, relationship to ultraviolet radiation and progression from benign to malignant tumors, may also exist in dogs. Canine and human mucosal melanomas appear to harbor BRAF, NRAS, and c-kit mutations uncommonly, compared with human cutaneous melanomas, although both species share AKT and MAPK signaling activation. We conclude that there is significant overlap in the clinical and histopathological features of canine and human mucosal melanomas. This represents opportunity to explore canine oral cavity melanoma as a preclinical model.

A primary intimal aortic angiosarcoma was diagnosed in a 4-year-old, spayed female German Shepherd that presented for complications of thromboembolic disease because of infarcts in multiple organs. On gross examination, aneurysmal dilatation of the aorta was associated with a friable, necrotic mass attached to the endothelial surface, which partially occluded the aortic lumen. On histologic examination, plump neoplastic spindle cells formed a plaque-like mass arising from the intima that merged with a large accumulation of fibrin and necrotic debris, and projected into the lumen. Neoplastic cells invaded periaortic vessels and were seen in some infarct-associated thromboemboli. Tumor cells expressed vimentin and CD31, with infrequent, patchy staining with factor VIII-related antigen; tumor cells were negative for cytokeratin and smooth-muscle actin. Aortic angiosarcoma is a rare malignancy in humans. This is the first description of a primary intimal aortic angiosarcoma in a dog, with immunohistochemical evidence of endothelial origin.

The elbow joint is one of the feline appendicular joints most commonly and severely affected by degenerative joint disease. The macroscopic and histopathological lesions of the elbow joints of 30 adult cats were evaluated immediately after euthanasia. Macroscopic evidence of degenerative joint disease was found in 22 of 30 cats (39 elbow joints) (73.33% cats; 65% elbow joints), and macroscopic cartilage erosion ranged from mild fibrillation to complete ulceration of the hyaline cartilage with exposure of the subchondral bone. Distribution of the lesions in the cartilage indicated the presence of medial compartment joint disease (most severe lesions located in the medial coronoid process of the ulna and medial humeral epicondyle). Synovitis scores were mild overall and correlated only weakly with macroscopic cartilage damage. Intra-articular osteochondral fragments either free or attached to the synovium were found in 10 joints. Macroscopic or histologic evidence of a fragmented coronoid process was not found even in those cases with intra-articular osteochondral fragments. Lesions observed in these animals are most consistent with synovial osteochondromatosis secondary to degenerative joint disease. The pathogenesis for the medial compartmentalization of these lesions has not been established, but a fragmented medial coronoid process or osteochondritis dissecans does not appear to play a role.

Human immunodeficiency virus (HIV) and ovine progressive pneumonia virus have been associated with lymphocytic pneumonitis. Pulmonary cell populations in cats infected with feline immunodeficiency virus (FIV) were evaluated by bronchoalveolar lavage (BAL) to identify changes associated with lentivirus infection in this species. Bronchoalveolar lavage was performed through an endotracheal tube using 15 ml kg−1 body weight of sterile 0.9% sodium chloride solution. Results of BAL fluid cytologic analysis from 19 cats experimentally infected with FIV for at least 8 months were compared with results from 34 uninfected cats. Infected cats had significantly higher total cell counts and relative neutrophil counts (P < 0.01). Lymphocytosis did not occur. Bronchoalveolar lavage fluid was collected from nine additional cats prior to, and 2, 6, and 17–18 weeks following infection with FIV. Neither neutrophilia nor lymphocytosis was associated with FIV infection in these cats.

Spirochetemia is a rarely reported observation in dogs. We describe the clinical, hematologic, and immunodiagnostic features of two spirochetemic dogs from northern Florida and the subsequent isolation and preliminary characterization of a Borrelia species from one dog in which culture of a sample for spirochetes was attempted. Results of polyacrylamide gel electrophoresis, monoclonal antibody testing, and PCR analysis indicate that the Florida isolate is not Borrelia burgdorferi, the only other member of the genus that has been isolated in Florida. Our findings also indicate that a member of the genus Borrelia potentially causes disease in dogs in Florida and that serologic cross-reactivity of the Florida canine Borrelia isolate with B. burgdorferi probably contributes to the inaccurate diagnosis of canine Lyme disease in the region.

2-Mercaptobenzimidazole (2-MBI) is a member of the thioureylene compound family known for their potent antithyroid activity. Male Fischer-344 rats were exposed to 0.0, 3.13, 12.5, and 50.0 mg/m3 of 2-MBI via inhalation for 13 wk. Follicular cell hyperplasia occurred in the thyroid glands of rats from the 3 2-MBI treatment groups. Thyrotrophs in the pituitary glands from rats in these 3 groups were hyperplastic, and they had varying numbers of hypertrophic cells with either eosinophilic stippled cytoplasm or with eosinophilic globules within 1 or more large vacuoles that displaced the nucleus. These cells were compared by immunohistochemistry and ultrastructural examination to "thyroidectomy cells" within the anterior pituitary glands of thyroparathyroidectomized rats and were determined to be identical to them. Immunohistochemical staining for the β-chain of thyroid-stimulating hormone confirmed that the hyperplastic and hypertrophic cells were thyrotrophs. Ultrastructurally, hypertrophic cells in 2-MBI-treated rats and thyroidectomy cells in thyroparathyroidectomized rats had expanded cytoplasm containing either increased profiles of rough endoplasmic reticulum (RER) or one or more widely dilated cisternae of RER, which displaced other cellular organelles. 2-MBI appears to act comparably to other thioureylene compounds that have been shown to produce low serum concentrations of triiodothyronine and thyroxine, which results in thyrotroph hypertrophy and hyperplasia, resultant thyroid hyperplasia, and subsequent goiter.

Morphologic and biologic features of five feline granulated round cell tumors were compared with those previously reported to be of globule leukocyte and large granular lymphocyte origin. The five cats ranged from 6 to 9 years of age and presented with nonspecific gastrointestinal signs. Four of the five cats were tested for feline leukemia virus and were negative by enzyme-linked immunosorbent assay. The neoplastic process involved the abdominal cavity in all cases, with a predilection for the distal small intestine and mesentery. The liver and peripheral and thoracic lymphoid tissues were also sporadically affected. Neoplastic round cells contained 0.5–1.5-μm eosinophilic cytoplasmic granules that were difficult to discern on casual observation with hematoxylin and eosin stain but were deep blue and easily visualized when stained with phosphotungstic acid-hematoxylin. In two cases, epithelium in the affected ileum and liver contained unusually large numbers of apparently normal globule leukocytes. Ultrastructurally, the tumor granules tended to cluster at one nuclear pole and were spindle to round in shape with variably dense contents. Some granules contained a dense “cap” at one end or internal crystalloid bars that distorted the granule membrane. The tumors reported herein are similar to all three of the previously reported feline granulated round cell tumors and probably have a common cellular origin.

Recently released results of a supply and demand survey (http://www.toxpath.org/acvp_workforce. asp) of veterinary pathologists conducted in 2007 by the American College of Veterinary Pathologists (ACVP), the American Society of Veterinary Clinical Pathology, and the Society of Toxicologic Pathology (STP) confirm the results of a similar survey conducted in 2002 (http://www.acvp.org/news/ 2002SupplyDemand/2002Supply.pdf)—there is a continued shortage of veterinary pathologists to fill a growing number of critical positions in academia, industry, and government. Comparison of the results of the 2 surveys, however, indicates that considerable progress has been made in the last 5 years, particularly on the supply (training) side. Specifically, it has become easier to recruit candidates for postDVM pathology training, the quality of the candidate pool has improved, and there are more positions and trainees in veterinary anatomic pathology and clinical pathology training programs. In response to the workforce shortage originally documented in the 2002 survey, the ACVP/STP Coalition for Veterinary Pathology Fellows was established to provide a unified mechanism to solicit and allocate funds to train additional, much-needed veterinary pathologists. After approximately 4 years of operations, this unique educational initiative has established partnerships between academia and the biopharmaceutical industry to create 22 new training positions, including 15 anatomic pathology residencies, 3 clinical pathology residencies, and 4 postresidency PhD pathology research positions. These positions were funded by Bristol-Myers Squibb, Charles River Laboratories, Eli Lilly, Genentech, GlaxoSmithKline, Merck, Pfizer, sanofi-aventis, Schering-Plough, and Wyeth. Importantly, sponsors who funded the initial 3-year training positions have since renewed funding for another set of positions. ‘‘The visionary participation of these world-class biopharmaceutical companies and contract research organizations and their commitment to the training of future veterinary pathologists attest to their recognition that data submitted by these companies to regulatory agencies must always be of the highest quality,’’ noted Dr. Ricardo Ochoa, president of the STP. ‘‘By assuring the supply of well-trained veterinary pathologists, some of whom will likely become employed in private industry, these organizations are planning for the continued accuracy of decisions on the safety and efficacy of new chemical entities in the future.’’ Sponsors provide support for stipend, tuition and fees, benefits, travel, and educational supplies for 3 years. In addition to support for the training positions themselves, other sponsors have supplied unrestricted contributions for important enabling activities of the Coalition. Companies that have offered this form of support include Allergan, Aperio, Bristol-Myers Squibb, Burroughs-Wellcome Fund, Elsevier, Experimental Pathology Laboratories, GEMpath, Histo-Scientific Research Laboratories, Integrated Laboratory Systems, Millennium-Takeda, The Leyden Group, and Wyeth. Taken together, $5.2 million in commitments has been raised during the past 4 years. At the conclusion of their training, Fellows have no payback obligation or commitment to sponsors other than to complete the ACVP certification examination and pursue a career in veterinary pathology. The new positions were awarded on a competitive application basis to 14 different North Vet Pathol 46:565–566 (2009) DOI: 10.1354/vp.09-VP-0036-C-COM

To determine whether it was possible to retrieve organisms, by means of bronchoalveolar lavage (BAL), from cats inoculated with Toxoplasma gondii.Experimental study.27 cats. Sixteen of the 27 were experimentally infected with feline immunodeficiency virus.All cats were inoculated with T gondii tachyzoites. Cats were grouped on the basis of feline immunodeficiency virus status and route (IV or intra-arterial) and number of tachyzoites administered. Bronchoalveolar lavage was performed by means of a standard technique. Lavage fluid was evaluated cytologically for tachyzoites.Clinical signs of toxoplasmosis varied widely among individual cats, but were generally most pronounced in group-1 and -2 cats (n = 5 each) and less pronounced in group-3 (n = 5) cats. Group-4 and -5 cats (n = 6 each) did not have clinical signs of toxoplasmosis. In 14 of the 15 cats in groups 1, 2, and 3, tachyzoites were detected in BAL fluid collected 7 days after inoculation. Tachyzoites were detected 14 days after inoculation in the single cat without tachyzoites 7 days after inoculation. A necropsy was performed on 9 of these cats, and tachyzoites were identified histologically in 4 of the 9. Tachyzoites were not detected in BAL fluid collected 3 days (n = 6) or 7 days (n = 6) after inoculation from the 12 cats in groups 4 and 5. Tachyzoites were not identified histologically in any of these 12 cats.BAL may be useful in the diagnosis of toxoplasmosis, particularly in cats with signs of pulmonary involvement.

To characterize the pathogenic potential of a unique Borrelia isolate obtained from a dog from Florida (FCB isolate).Prospective experimental infection.32 preweanling Swiss Webster mice and 12 adult male Hartley guinea pigs were injected intraperitoneally with 10(5) spirochetes.Mice were used as controls and blood recipients, and at 3- to 4-day intervals, 1 control mouse and 2 infected mice were necropsied, tissues were cultured, and a recipient mouse was inoculated with blood. Guinea pigs were randomized to 4 groups and inoculated intradermally with 10(0), 10(2), 10(3), or 10(4) spirochetes. For 48 days, clinical, hematologic, serologic, and microbiologic tests were performed on them, after which they were necropsied.In mice, spirochetemia was detectable between postinoculation days (PID) 3 and 13, and seroreactivity to homologous antigen was detectable during PID 10 through 31. Compared with control mice, infected mouse spleens were 2 to 3 times larger. Histologic lesions included lymphoid hyperplasia, neutrophilic panniculitis, epicarditis, and myocarditis, with intralesional spirochetes detected from PID 3 through 6. During PID 10 through 31, nonsuppurative epicarditis developed. Signs of illness and hematologic abnormalities were not observed in guinea pigs, despite isolating spirochetes from blood during PID 7 to 27. When necropsied on PID 48, histologic lesions included lymphoid hyperplasia and lymphocytic plasmacytic epicarditis.The FCB isolate causes spirochetemia, lymphoid hyperplasia, dermatitis, and myocardial injury in Swiss Webster mice and can be transmitted by blood inoculation. In Hartley guinea pigs, the isolate causes spirochetemia, lymphoid hyperplasia, and epicarditis. Documentation of disease in mice, guinea pigs, and, presumably, dogs raises the level of concern that the FCB isolate might be pathogenic for man and other animal species.

The objectives of this study were to determine in horses 1) the ages at which viable cartilage canal vessels are present in the articular–epiphyseal cartilage complex of three predilection sites and one nonpredilection site of osteochondrosis (OC), 2) the prevalence of lesions of OC in these sites, and 3) whether there was an association of lesions of OC with necrotic cartilage canal blood vessels. The medial femoral condyle, lateral femoral trochlear ridge, and distal ends of the tibia and proximal phalanx were examined grossly, microradiographically, and histologically in 35 horses 18 months old or younger. Cartilage canals containing patent blood vessels were present in all sites examined in foals less than 3 weeks old and were absent from all sites by 7 months of age. The overall prevalence of lesions of OC at one or more of the sites examined was 12/35 (34%). Prevalence increased to 11/22 (50%) in horses 2 months old and older. These lesions occurred primarily in the medial condyle of the femur ( n = 5) and the intermediate ridge of the distal tibia ( n = 5). All lesions seen in horses between 3 weeks and 5 months of age were associated with necrotic cartilage canal blood vessels. In horses 7 months of age and older, lesions of OC were considered chronic because of extensive involvement of the subchondral bone and bone marrow. These results suggest that OC lesions develop prior to 7 months of age and that ischemic necrosis of cartilage secondary to a defect in vascular supply is an important factor in the pathogenesis of this disease in horses.

Hypercalcemia was found in a dog examined for episodic weakness and disorientation. A right adrenal mass identified on abdominal ultrasonographic examination was diagnosed as a pheochromocytoma based on increased 1231-metaiodobenzylguanidine uptake and characteristic histopathological features. Serum intact parathyroid hormone (iPTH), total serum calcium, and ionized serum calcium were increased prior to surgery. Although both total and ionized serum calcium concentrations decreased to within normal reference ranges following surgery through postoperative day 17, subsequent high concentrations with increased serum iPTH concentrations confirmed concurrent primary hyperparathyroidism. A right parathyroid tumor was suspected based on a technetium-99m-sestamibi (99mTc-sestamibi) scan.

A Comparison Between Surgical and Standing Percutaneous Contact Nd:YAG LaserPeriosteal Transection for Potential Treatment of Equine Limb Deformities.L. P. Tate, S. J. Baines, D. J. Meuten, J. StefanacciNorth Carolina State UniversityDepartment of Food Animal and Equine MedicineCollege of Veterinary Medicine, Raleigh, North Carolina 27606ABSTRACTMedial and lateral aspects of both radiuses were subjected to periostealtransection of 9 healthy equine foals. One site per foal was subjected tosurgical periosteal transection and elevation under general anesthesia. Theremaining 3 sites of each foal after injection of a local anesthetic receivedNd:YAG contact percutaneous periosteal transection. All radiuses were evaluatedradiographically prior to periosteal transection and immediately prior toeuthanasia. Foals were euthanized at 3 days, 31 to 34 days, and 67 days post-periosteal transection and gross postmortem and histologic examination performedon each site.Radiographically, periosteal proliferation occurred at all the conventionalsurgery sites and a majority of the percutaneous laser sites by 30 days post-treatment and was present at 67 days. No limb angulations were noted to occur inany of the foals.The histologic appearance of the conventional surgery site demonstrated aproliferative layer of periosteum which was thickened and disorganized. Lasersite differed in that they were surrounded by a zone of cortical bone whichappeared grossly normal, but osteocytes were absent from the lacunae. Theconclusion of the study was that Nd:YAG percutaneous laser periosteal transectionwas successful in producing periosteal obliteration but with a differenthistologic appearance than that produced by conventional surgery. The findingsdid support the hypothesis that standing laser periosteal transection maysubstitute for general anesthesia and conventional periosteal transection andelevation for treatment of angular limb deformities in foals.

Bronchoalveolar lavage was performed through an endotracheal tube in 34 specific-pathogen-free cats to determine expected values for bronchoalveolar lavage fluid cytologic analysis, using this method of collection. Saline solution for lavage was instilled in 3 separate aliquots at a volume of 5 ml/kg of body weight each. Analysis of sequential aliquots was performed to investigate the differences in cell counts among the 3 fractions. The effect of combining aliquots, including or omitting the first fraction, was evaluated to determine whether all aliquots could be combined for analysis without substantially affecting results. The total number of nucleated cells retrieved from each cat ranged from 0.9 to 31.1 x 10(6). Most of these cells were macrophages (78 +/- 15%, mean +/- SD) and eosinophils (16 +/- 14%). The first aliquot had the greatest number of epithelial cells, and the lowest total nucleated cell count and relative and absolute eosinophil counts. Differences among aliquots also were identified for relative and absolute macrophage counts, relative and absolute neutrophil counts, and absolute lymphocyte count. Statistically significant differences were found for many of the cell counts when values from the combination of the second and third aliquots were compared with values from the combination of all 3 aliquots. Magnitude of the differences was small, and these differences were not believed to be of practical consequence.

Visceral leishmaniasis was diagnosed by cytology and positive indirect immunofluorescent antibody titers to Leishmania donovani in a 7-month-old female Basenji dog from Texas. Clinical and laboratory findings included weight loss, hematochezia, hyperglobulinemia, hypoalbuminemia, anemia, and neutrophilic leukocytosis. Evidence of response to treatment with diminazene aceturate and ketoconazole included improvement in the abnormal clinical, hematologic, and biochemical findings, decreased serum globulin concentration and antibody titer to Leishmania donovani, and absence of organisms in examined tissues. Several foci of endemic leishmaniasis have been reported in the United States. Because of its zoonotic potential and the lack of approved treatments for dogs with leishmaniasis in the United States, the development of effective treatment strategies is needed.

Summary Specialized training is required to obtain adequate cytology smears by needle aspiration biopsy. An educational method that would provide opportunity for numerous trainees to acquire needle aspiration biopsy and cytodiagnostic skills, and which would not result in the interruption of teaching hospital patient care nor result in the use or sacrifice of animals from laboratory animal resources was sought. An aspiration biopsy cytology teaching aid was developed by placing portions of fresh organ tissue from routine submissions to the necropsy laboratory into a specimen container partially filled with chilled normal saline solution. Specimens were stored immersed in saline at 4°C for up to 48 hours. Prior to the cytopathology teaching laboratory period, the specimen container was removed from storage and covered with a latex diaphragm. Instructors responsible for teaching cytodiagnostic techniques used this ex vivo aspiration biopsy cytology device for instruction of fine needle aspiration biopsy, specimen processing, and microscopy. In the teaching laboratory, puncture of the diaphragm and aspiration of a tissue sample with hypodermic needle and syringe was instructed to, and practiced by, numerous trainees simultaneously using clinical operative technique without the need for live animals. Trainees prepared and stained cytology smears as well as evaluated and interpreted specimens in a single, realistic exercise. Such training experience may improve ability of trainees to procure diagnostic-quality biopsy specimens for cytodiagnosis of lesions from clinic patients.

This study was designed to examine the association of spontaneous lesions of osteochondrosis with vascular supply to epiphyseal cartilage, and to determine whether similar lesions could be experimentally reproduced by selective interruption of cartilage canal blood supply. The vascular supply to the articular-epiphyseal cartilage complex of the distal end of the femur was studied in 27 microfil- or barium-injected and cleared specimens and 24 serially sectioned microangiographic specimens from 27 clinically normal female swine (3.6 to 71.0 kg). Blood vessels supplying the articular-epiphyseal cartilage complex were consistently restricted to the epiphyseal region and the number of vessels decreased as the pigs increased in weight (p less than 0.001). Spontaneous lesions of osteochondrosis (i.e., cartilage necrosis) were initially seen in the first areas of epiphyseal cartilage to become avascular and were associated with necrotic blood vessels. The number and size of foci of necrotic cartilage increased as the pigs increased in weight (p less than 0.001). Blood supply to epiphyseal cartilage from cartilage canal vessels was surgically interrupted in a highly vascular area of the medial femoral condyle in eight additional 23-kg female swine. This procedure resulted in necrosis of blood vessels within cartilage canals followed by necrosis of surrounding cartilage, lesions that appeared to be identical to early spontaneous lesions of osteochondrosis. These results suggest that the viability of epiphyseal cartilage in the articular-epiphyseal cartilage complex is highly dependent on an adequate blood supply from cartilage canal vessels, and strongly implicates a defect in blood supply in the pathogenesis of osteochondrosis.

A 12-year-old kitten was found to have bone fragility. Previously described causes of bone fragility in domestic cats were excluded on the basis of serum biochemical profile and radiographic histologic examinations. The findings in this kitten correlated most closely with osteogenesis imperfecta, a syndrome documented in human beings and cattle.

Intestinal morphology and fluid and electrolyte transport were examined in a neonatal porcine model of cryptosporidiosis. Sections of jejunum, ileum, and colon were obtained for morphometric analysis on days 3, 6, 9, and 12 postinfection, and in vivo perfusion studies of jejunum and ileum were conducted on days 3 and 4 postinfection. The most severe morphologic lesion was seen in the ileum on day 3, and consisted of villous atrophy, crypt hyperplasia, and cellular infiltration. Villous surface area was reduced from 2.1 +/- 0.4 x 10(5) microns2 in control ileum to 0.8 +/- 0.1 x 10(5) microns2 in infected ileum, a result associated with enterocytes that were fewer in number and reduced in cross-sectional area. Conversely, the number of inflammatory cells in the lamina propria of the villus increased from 456 +/- 116 in control to 1014 +/- 187 in infected villus without a significant change in the volume of the lamina propria. At the height of infection, there was an approximate 1:2 ratio of both organisms and inflammatory cells to villous enterocytes. In contrast, organisms were not observed in the crypts, and the concentration of inflammatory cells in crypt lamina propria was unaltered. Disappearance of organisms and polymorphonuclear cells from the ileum was associated with restoration of normal structure and was complete by day 12. Although organisms were seen in the colon, the general architecture was not severely affected. On days 3 and 4 postinfection, there was a complete impairment of the glucose-stimulated Na and water absorption in both jejunum and ileum of infected pigs; however, absorption of electrolytes and water from a basic Ringer's solution, in the absence of glucose, was not significantly affected. These results are consistent with a malabsorptive diarrheal disease associated with the morphological damage and are very similar to those seen in enteric viral disease in pigs, except that the upper intestine is more severely affected in the latter.

Lipogranulomatous lymphangitis of the intestine occurred in four dogs with intestinal lymphangiectasia. All four presented with chronic diarrhea; three had ascites and two had hypoalbuminemia. Lipogranulomas appeared in lymphatics, often at the point of mesenteric attachment, of small intestine, ileum, or ileum and colon. Mesenteric lymphatics were obstructed and villous lacteals were distended. Mesenteric lymph nodes of one dog contained large lipid spaces and that same animal had a solitary subcapsular lipogranuloma of the liver. This disorder is one of several that result in protein-losing enteropathy in dogs.

Multinucleated syncytial cells consisting of clusters of closely apposed, darkly staining nuclei and densely eosinophilic cytoplasm were present in parathyroid glands from 25 of 36 dogs (69%). They were most often located either peripherally in the parathyroid gland or adjacent to blood vessels. The numbers of nuclei per syncytial cell ranged from two to greater than 30. Syncytial cells had degenerative changes as suggested by vacuolated mitochondria, dilated rough endoplasmic reticulum, irregular nuclear outlines, and condensed nuclear chromatin. These ultrastructural changes contrasted sharply with adjacent chief cells that had well-preserved organelles. The presence of desmosomes and fragments of cell membranes between nuclei suggested that syncytial cells were derived from fusion of adjacent chief cells rather than nuclear division without cytokinesis. Syncytial cells were present in normal, hyperplastic, and inactive parathyroid glands from dogs. There were no significant correlations between the presence of multinucleated cells and age of dog, parathyroid histology, and changes in the concentrations of serum calcium, phosphorus, creatinine, or plasma parathyroid hormone. Syncytial cells occurred frequently in canine parathyroid glands regardless of the state of activity of the gland or the concentration of plasma parathyroid hormone or serum calcium.

Articular-epiphyseal cartilage complexes from the distal femur and humerus of five 3-month-old pigs were collected and preserved using either a conventional or a ruthenium hexammine trichloride (RHT)-supplemented fixation technique. Lesions were similar regardless of the fixation technique. Areas of necrosis were in epiphyseal but not articular cartilage from both sites of all pigs. Cartilage canals were confined to epiphyseal cartilage and contained vessels which had endothelial cells in varying stages of degeneration and necrosis. Areas of necrotic cartilage often were adjacent to or surrounded degenerate cartilage canals. Lipid emboli (up to 40.0 γm in diameter) were infrequently located in vessels within cartilage canals. Associated with the lipid emboli were leukocytes, erythrocytes, necrotic cell remnants, and flocculent material. Restriction of necrosis to epiphyseal cartilage and the association of these necrotic areas with degenerating vessels in cartilage canals strongly implicate a defect in cartilage canal blood supply in the pathogenesis of osteochondrosis. The RHT fixation technique resulted in excellent cellular detail at the light microscopic level, but ultrastructurally there was marked vacuolation of chondrocytes and matrix. The conventional technique caused shrinkage of all chondrocytes, resulting in a wide halo of pericellular matrix surrounding each cell.

An adult Keeshond had clinical signs associated with hypercalcemia, including inappetence, polyuria, polydipsia, and vomiting. Blood biochemical findings and urinary clearance studies were consistent with a diagnosis of primary hyperparathyroidism. Histomorphometric analysis of trabecular bone in an iliac crest biopsy indicated increased bone remodeling activity. Surgical exploration of the neck revealed an oval mass, which was removed by blunt dissection. Histologic diagnosis was parathyroid gland adenoma. The dog died because of renal failure on the eighth postoperative day. This report defines primary hyperparathyroidism in the dog, thus facilitating diagnosis for the veterinary clinician.

Mice given propylthiouracil, a thyroid inhibitor, and fed a diet containing a nontoxic level of rac-1(3)-palmitoyl glycerol showed the hypothermia and mortality expected for a toxic dose, but did not show these signs when linoleate or oleate was added to the diet. Loss of radioiodine from the whole animal and thyroid gland was slower when mice were fed the toxic palmitoyl glycerol diet than when fed the same diet containing 4% safflower oil. However, mice fed the two diets did not differ in the extent of the incorporation of radioiodine, and essentially all was bound to protein in each case. Follicular thyroid cells from mice fed the potentially toxic diet that contained unsaturated fat were normal in appearance. Conversely, cells from mice fed the toxic diet were smaller and more densely stained, showing evidence of glycoprotein inside the cell. These findings show that the thyroid gland is affected by the palmitoyl glycerol diet. However, the thyroid is not the only organ affected, because giving either thyroxine or triiodothyronine had no effect on the toxicity of palmitoyl glycerol.

Adenocarcinomas derived from apocrine glands of the anal sac and associated with persistent hypercalcemia in dogs were composed of tumor cells with numerous profiles of rough endoplasmic reticulum, clusters of free ribosomes, and a prominent Golgi apparatus. Neoplastic cells contained microtubules, microfilaments, tonofibrils, and had two types of electron-dense granules. Large lysosomelike dense bodies ranged from 0.6 to 2.2 microns in diameter and had a poorly delineated limiting membrane. Small granules (150-400 nm in diameter) had a sharply delineated limiting membrane with a narrow submembranous space and a homogeneous dense core. These smaller granules usually were located near the apexes of neoplastic cells, whereas the larger granules were situated near the base of cells. Apocrine cells in glands of the anal sac from control dogs that were in the secretory phase were columnar and had large dilated profiles of rough endoplasmic reticulum. Membranes of the endoplasmic reticulum fused with the plasmalemma and appeared to secrete their product directly into the lumens of acini, characteristic of merocrine secretion. Apical blebs of electron-lucent cytoplasm pinched off from nonneoplastic aprocine cells and were released into glandular lumens. Similar electron-lucent cytoplasmic blebs were present at the apexes of tumor cells. Myoepithelial cells were present between the epithelial cells and basement membrane in normal apocrine glands and were absent in neoplasms derived from these glands. Identification of the contents of the secretory-like granules in tumor cells and characterization of the hypercalcemic factor in the plasma or tumor tissue from dogs with this syndrome will help explain the pathogenesis of hypercalcemia associated with malignancy in animals and man.

Osteochondrosis of the articular-epiphyseal cartilage complex (A-E complex) is a significant clinical disease in swine. It has been suggested that osteochondrosis is the underlying cause of osteochondritis dissecans in humans. The purpose of this investigation was to characterize the ultrastructural changes in the earliest macroscopically visible lesion of the epiphyseal cartilage in osteochondrosis of the A-E complex in swine. Osteochondritic epiphyseal cartilage from the distal femora and humeri of growing crossbred boars was collected, embedded in plastic, and studied light and electron microscopically. The predominant lesion was chondronecrosis, characterized by chondrocyte death and loss of matrical proteoglycan. Transition from normal to abnormal cartilage was abrupt. Lipid accumulated in chondrocytes within and adjacent to lesions, but not in chondrocytes distant from lesions. Intracellular lipid accumulation was an important feature of the lesion and may play a role in its initiation. It is hypothesized that intracellular lipid accumulation results from hypoxia/anoxia and may precede matrix degeneration, which precedes cell death.

Malignant histiocytosis was diagnosed in a 4-year-old-dog. Major clinicopathologic findings included multiple cutaneous tumors, peripheral lymphadenopathy, hepatomegaly, lytic bone lesions, anemia, thrombocytopenia, and monoclonal IgA gammopathy. Pathologic findings included multi-systemic infiltration of erythrophagocytic malignant histiocytes. Chemotherapy was unsuccessful.

Chronic granulomatous enteritis associated with weight loss and hypoproteinemia was identified in 2 horses. Both horses continued to have normally formed feces. Malabsorption of carbohydrate and lipid, with concomitant gastrointestinal protein loss was demonstrated in 1 case. One horse was treated symptomatically and gained 108 kg. In both cases, principal gastrointestinal lesions were partial to total villus atrophy and transmural mononuclear leukocytosis, with lymphocytes and histiocytes predominating. The cause of the condition was not identified in either case.

Primary hyperparathyroidism was diagnosed in two German shepherd pups from a litter of four females. Clinical signs were apparent by two weeks of age and included stunted growth, muscular weakness, and polydipsia/polyuria. Radiography revealed diffuse reduction in bone density. Both pups had marked hypercalcemia, hypophosphatemia, increased plasma immunoreactive parathyroid hormone concentrations and increased fractional clearance of inorganic phosphate in the urine. Intravenous infusion of one affected pup with calcium gluconate failed to suppress the plasma concentration of immunoreactive parathyroid hormone, suggesting autonomous secretion of parathyroid hormone. Necropsy of the other pup at eight weeks of age revealed diffuse hyperplasia of parathyroid chief cells, nodular hyperplasia of thyroid C-cells, skeletal alterations consistent with fibrous osteodystrophy, hypercalcemic nephropathy, and extensive mineralization of the lungs and gastric mucosa. The dam and sire were half sibs. One male pup from a previous litter of six had developed similar clinical signs and radiographic lesions, suggesting autosomal recessive inheritance. This is the first report of hereditary primary hyperparathyroidism in domestic animals, a disease which may be analogous to hereditary neonatal primary hyperparathyroidism in children.

Three canine laryngeal tumors were diagnosed as oncocytomas by light microscopy, but were determined to be rhabdomyomas following ultrastructural and immunocytochemical examination. Tumors consisted of large eosinophilic cells interspersed with smaller dark cells. Large tumor cells had a granular, intensely eosinophilic cytoplasm. Scattered through the tumors were a few elongated cells with cytoplasmic cross striations and multiple nuclei. Tumor cells from all three dogs contained numerous mitochondria and bundles of myofibrils with electron-dense Z-lines typical of striated muscle cells. Intracellular myoglobin and desmin were detected in the tumors by immunocytochemistry. Comparisons are made with a previous report of canine laryngeal oncocytomas.

Rhinosporidiosis was diagnosed in six dogs from the southeastern United States. All six dogs had unilateral nasal polyps with multiple small white sporangia visible beneath the surface. Microscopically, the polyps consisted of organisms and fibrovascular tissue with a surface of columnar or squamous epithelium. Juvenile sporangia were unilamellar, 15-75 in diameter, nucleated, and accounted for about 65% of sporangia seen. Approximately 5% of the sporangia were in intermediate stages of maturation, were bilamellar, 100-150 μm in diameter, and contained immature endospores. Mature sporangia comprised about 30% of the total, were usually unilamellar, 100-400 μm in diameter, and contained a mixture of immature and mature endospores. The inner layer of the wall of the intermediate sporangia and the single wall of the mature sporangia were argyrophilic and carminophilic. Ultrastructurally. the earliest stage contained a nucleus and many ribosomes, lipid droplets, and phagolysosomes. Maturing sporangia contained discrete membrane-bound, round clevage products. These structures subsequently matured to spores, each of which had a wall and contained a nucleus and many lipid droplets. The organism from one dog was cultured and grown in vitro for 7 months and is the first successful cultivation of Rhinosporidium seeberi.

Hypercalcemia, hypercalciuria, and hyperphosphaturia were present in female dogs with adenocarcinomas derived from apocrine glands of the anal sac (CA). Remission of hypercalcemia accompanied tumor excision in all six dogs undergoing surgery, whereas tumor recurrence or growth of metastases was associated with a return of hypercalcemia. Preoperatively, the plasma concentrations of immunoreactive parathyroid hormone in all dogs were undetectable or in the low normal range. Plasma concentrations of 13,14-dihydro-15-keto-prostaglandin E2 (PGE2M) and serum 1,25-dihydroxyvitamin D were not significantly different from control dogs. Urinary cyclic AMP and hydroxyproline were increased in dogs with CA. No immunoreactive parathyroid hormone was detected in extracts from tumor tissue, and parathyroid glands from dogs with CA had ultrastructural characteristics of secretory inactivity. Lumbar vertebrae from hypercalcemic dogs had decreased trabecular bone volume and increased osteoclastic bone resorption compared with age-matched control dogs. After tumor excision, serum total calcium returned to the normal range, whereas immunoreactive parathyroid hormone increased 2- to 20-fold and 1,25-dihydroxyvitamin D decreased 2- to 8-fold. Postoperative hypocalcemia was not observed. These results indicate that CA produces a hypercalcemic factor other than immunoreactive parathyroid hormone or prostaglandin E2 that increases osteoclastic osteolysis distant from the tumor and results in hypercalcemia, hypercalciuria, and hyperphosphaturia.

Within a period of 5 consecutive days after the initial observation of illness was made, 7 of 12 Siberian Husky dogs developed clinical signs of Rickettsia rickettsii infection. One dog died and was necropsied. Clinical signs of infection consisted of lethargy, anorexia, ocular and nasal discharges, and neurologic disorder (incoordination and rolling). Scleral blood vessel injection, fever, lymphadenomegaly, splenomegaly, and increased bronchovesicular lung sounds were prominent findings. Clinical laboratory test results identified decreased platelet numbers, variable neutrophil counts, increased serum alkaline phosphatase activity, hyponatremia, hypokalemia, and bilirubinuria. Diagnosis of Rocky Mountain spotted fever was confirmed by serologic evaluation of acute and convalescent sera, using the micro-immunofluorescence technique, and R rickettsii antigen was determined by demonstration of intracellular rickettsial organisms in vascular endothelial cells of brain and lung (stained with carbol-basic fuchsin and aqueous malachite green) and by demonstration of spotted fever-group rickettsiae in tissues by direct fluorescent antibody technique. Near-simultaneous naturally occurring tick-borne infection of 7 dogs with R rickettsii documents an unreported occurrence.

Ninety-six recently weaned gilts were assigned to 3 groups. Group-1 gilts were fed a corn-soybean meal diet ad libitum and were killed when they weighed 110 kg. Gilts in groups 2 and 3 were fed 70% of the feed consumed by group-1 gilts. Group-2 gilts were killed when they weighed 110 kg, and group-3 gilts were killed at the same age as group-1 gilts. At slaughter, the right pelvic and thoracic limbs from all gilts were removed, and the distal end of each femur and humerus was sectioned serially at 5-mm intervals. Articular-epiphyseal cartilage complexes from these sites were examined grossly, radiographically, and histologically. Effects of treatment (ad libitum vs limited energy intake) and average daily gain on the prevalence and severity of lesions were analyzed statistically, using chi 2 analysis, a categoric-modeling technique, analysis of variance, and correlation coefficients. Of humeri and femora, 49 and 70%, respectively, had lesions of osteochondrosis, and 86% of the gilts had a lesion at one or both sites. Group-2 gilts (killed at 110 kg, but approx 70 days older than group-1 gilts) had significantly lower prevalence (P = 0.009) and severity (P = 0.002) of osteochondrosis of the distal end of the femur than did gilts in groups 1 and 3. When each group was evaluated individually, gilts that had a higher average daily gain had an increased prevalence and severity of lesions.

Pseudohyperparathyroidism was diagnosed in a mature stallion presented for anorexia, weight loss, pollakiuria and constipation. Laboratory findings included hypercalcemia, hypophosphatemia, anemia and isosthenuria. Thoracocentesis indicated an exfoliating squamous cell carcinoma. At necropsy, a squamous cell carcinoma of the stomach with metastases to the abdominal and thoracic cavities was diagnosed. No osseous metastases were found. No gross or microscopic renal lesions were noted. Bone tissue showed arrested resorption, and the parathyroid gland was atrophic.

Idiopathic hyperaldosteronism was diagnosed in an 8-year-old castrated male Yorkshire Terrier, based on increased concentration of plasma aldosterone, hypertension, hypernatremia, decreased natriuresis, hypokalemia, and hyperkaluria. Unilateral adrenalectomy was performed after visualization of a nodule on the right adrenal gland. Hyperplasia of the zona glomerulosa and increased postoperative aldosterone concentrations supported the diagnosis of idiopathic hyperaldosteronism.

Clinical, gross, and light microscopic findings are described for 36 dogs, 33 females and three males, with adenocarcinomas arising from the apocrine glands of the anal sac. All tumors had light microscopic features of malignancy and 22 of 23 metastasized to iliac and lumbar lymph nodes. Nine dogs had disseminated metastases, but bone metastases were found in only one dog. Differentiated neoplasms formed secretory acini and tubules lined by tall columnar or cuboidal epithelium. Most neoplasms were histologically bimorphic, with glandular areas and solid nests. Parathyroid glands were atrophic. Hypercalcemia (mean = 16.1 mg/dl) was present in 20 of 22 dogs (90%) and hypophosphatemia (mean = 3.2 mg/dl) in 12 of 17 (71%). Remission of hypercalcemia by tumor ablation and recurrence of hypercalcemia with tumor regrowth suggested that the tumor produced a substance that caused hypercalcemia. This unique clinicopathologic syndrome is characterized by hypercalcemia in old. predominantly female, dogs with an adenocarcinoma arising from the apocrine glands of the anal sac.

Rhinosporidium seeberi, a fungus that is associated with polyp-like tumors in animals and man, was successfully cultivated. This organism stimulated proliferation of epithelial cells in vitro, producing polyp-like structures. Spores produced in culture required a period of aging or development, or both, before they were capable of reinitiating the growth cycle.

Primary hypoparathyroidism was diagnosed in six dogs with profound hypocalcemia. Muscle tremors, tetany, generalized seizures, ataxia, and behavioral aberrations were the most common clinical signs. Lymphocytic parathyroiditis was found in four of five dogs that were biopsied. The serum concentration of immunoreactive parathyroid hormone was abnormally low in one dog in which it was measured. Treatment with vitamin D and calcium was successful in restoring and maintaining normal concentration of serum calcium in all six dogs. During treatment, large daily doses of vitamin D were required, dose response was unpredictable, serum calcium concentration fluctuated while treatment remained unchanged, and hypercalcemia occurred frequently.