Veterinary Pathology

Eighteen young Beagle dogs (eight males and 10 females), ages 6-40 months, with canine juvenile polyarteritis syndrome (CJPS), a naturally occurring vasculitis and perivasculitis of unknown etiology, were necropsied, and their tissues were examined by histopathologic and histochemical methods. The condition is characterized by recurring episodes of an acute onset of fever (>40 C) and neck pain that persist for 3-7 days. The major histopathologic alterations were a systemic vasculitis and perivasculitis. During the febrile, painful period of CJPS, the vascular lesions ranged from a histiocytic-lymphocytic periarterial infiltration to transmural arterial inflammation with concomitant fibrinoid necrosis and vascular thrombosis. Massive periarterial accumulations of inflammatory cells were common and often extended into adjacent tissues. The small- to medium- sized muscular arteries of the heart, cranial mediastinum, and cervical spinal meninges were consistently involved. Vasculitis occasionally occurred in other organ systems. The vascular lesions in dogs examined during clinically normal periods consisted of intimai and medial fibrosis, ruptured elastic laminae, and mild perivasculitis; these lesions were probably related to previous episodes of vasculitis. Eight dogs that had experienced repeated acute episodes also developed splenic, hepatic, and renal amyloidosis. The clinical signs, laboratory abnormalities, and the vascular lesions suggest that the condition may be immune-system mediated. CJPS may serve as a naturally occurring animal model of human immune-system-mediated vasculitides such as polyarteritis nodosa, infantile polyarteritis, and Kawasaki disease.

Following the Exxon Valdez oil spill, 347 oiled sea otters (Enhydra lutris) were treated in rehabilitation centers. Of these, 116 died, 94 within 10 days of presentation. Clinical records of 21 otters dying during the first 10 days of rehabilitation were reviewed to define the laboratory abnormalities and clinical syndromes associated with these unexpected deaths. The most common terminal syndrome was shock characterized by hypothermia, lethargy, and often hemorrhagic diarrhea. In heavily and moderately oiled otters, shock developed within 48 hours of initial presentation, whereas in lightly oiled otters shock generally occurred during the second week of captivity. Accompanying laboratory abnormalities included leukopenia with increased numbers of immature neutrophils (degenerative left shift), lymphopenia, anemia, azotemia (primarily prerenal), hyperkalemia, hypoproteinemia/hypoalbuminemia, elevations of serum transaminases, and hypoglycemia. Shock associated with hemorrhagic diarrhea probably occurred either as a direct primary effect of oiling or as an indirect effect secondary to confinement and handling in the rehabilitation centers. Lightly oiled otters were less likely to die from shock than were heavily oiled otters (22% vs. 72%, respectively). Heavily oiled otters developed shock more rapidly and had greater numbers of laboratory abnormalities, suggesting that exposure to oil was an important contributing factor.

Thirteen adult mutant (LWC strain) Japanese quails (Coturnix coturnix japonica), between the ages of 8 and 60 weeks were examined for a progressive muscular disorder. The disorder, inherited as an autosomal dominant trait, was clinically apparent as early as 28 days of age; it was characterized by generalized myotonia, muscle stiffness, and muscle weakness. Affected birds were identified by their inability to lift their wings vertically upward and by their inability to right themselves when placed on their dorsum. Electromyographic studies in two mutant quails showed high-frequency repetitive discharges comparable to those of myotonic runs. These discharges persisted after nerve resection. The distinctive histopathologic changes in the various muscles examined were ring fibers, sarcoplasmic masses, and internal migration of sarcolemmal nuclei. A slight decrease in the size of type IIB muscle fibers and a slight increase in the size of type IIA fibers were observed in the M. pectoralis thoracicus of affected quails. In older affected birds, inter- and intrafascicular fatty infiltration with replacement of type IIB fibers by fat cells was seen in the pectoral muscles. Single fiber necrosis, nonspecific lymphorrages, and variations in the muscle fiber size and shape were also noted. The typical muscle lesions and multisystem involvement, which was manifested by testicular degeneration and atrophy in the male LWC specimens and bilateral lenticular cataracts in 6 of 13 affected mutant quails, suggest resemblance of this new inherited muscular disorder to myotonic dystrophy in man.

The pathogenesis of porcine reproductive and respiratory syndrome virus (PRRSV) was determined in gnotobiotic pigs by studying the sequential development of microscopic lesions and sites of virus distribution and replication. Thirty-two pigs (three pigs/infected group and one pig/control group) were inoculated by nasal instillation of either PRRSV isolate ATCC VR-2332 (total dose 10^2,6 TCID₅₀) or uninfected cell culture supernatant. Infected and control pigs were euthanized at 12 hours, and 1, 2, 3, 5, 7, 14, and 21 days postexposure (PE). Gnotobiotic pigs experimentally infected with PRRSV were viremic by 12 hours PE and subsequently developed pneumonia, lymphadenopathy, vasculitis, myocarditis, and encephalitis. Lung lesions developed by day 3 PE, persisted through day 21 PE and were characterized by alveolar septa thickened by macrophages, alveolar proteinaceous and karyorrhectic debris, alveolar syncytial cells, and multifocal type II pneumocyte hypertrophy. Lymph node lesions varied in distribution and severity and were characterized by germinal center hypertrophy and hyperplasia, lymphocyte necrosis, multiple cystic spaces, and polykaryocytes within the cystic spaces. Heart lesions were a late feature of infection and all infected pigs had heart lesions on day 21 PE characterized by subendocardial, myocardial, and perivascular foci of lymphocytes. Vasculitis also varied in distribution and severity and affected all sizes of vessels. Results of this experiment indicate that PRRSV is a multisystem disease characterized initially by viremia with subsequent virus distribution and replication in multiple organs causing interstitial pneumonia, vasculitis, lymphadenopathy, myocarditis, and encephalitis.

A severe persistent neutropenia developed in a rabbit that was injected intradermally with 120, 60, 60, and 120 μg of recombinant canine granulocyte colony-stimulating factor (cG-CSF) on days 1, 22, 31, and 44, respectively. The neutropenia was present from day 44 to day 205. The nadir of the neutropenia (60 cells/μl) occurred in conjunction with peak antibody titer (640,000) to cG-CSF on day 58. The immune antiserum from this rabbit reacted positively for cG-CSF on Western blot analysis. The immune antiserum also neutralized the activity of cG-CSF. On day 160, examination of the bone marrow showed marked granulocytic hypoplasia and mild erythroid hyperplasia. On day 205, the rabbit was still neutropenic (430 cells/μl), even though the last injection of cG-CSF was given 161 days previously. Necropsy on day 205 showed that there was still mild granulocytic hypoplasia with mild erythroid hyperplasia. Because of the lack of any inflammatory foci found at necropsy and the granulocytic hypoplasia, it was thought that the neutropenia was most likely due to decreased production and was not a consumptive process. It is hypothesized that the antibody that was produced to cG-CSF neutralized the effect of endogenous rabbit granulocyte colony-stimulating factor and prevented the normal proliferation and maturation of the rabbit neutrophils.

Long-Evans (Eker) rats carry a mutation that predisposes them to develop spontaneous renal cell tumors of two morphologic patterns: solid chromophilic masses or cystic lesions lined by eosinophilic cells. Previous studies have suggested that these tumors arise from the proximal tubules. In the present study, lectin-binding characteristics and cytokeratin expression of various stages of hereditary rat renal epithelial neoplasia were examined to localize the portion of the nephron from which tumors arise. Lectin-binding histochemistry has been used as a marker of cell surface glycoprotein expression, thought to be important in the differentiation of benign from malignant epithelial lesions and in the determination of their cell of origin. The presence or absence of keratin intermediate filaments in the rat nephron has been used to identify nephron segments. The polyclonal antibody to high- and low-molecular-weight cytokeratin stained the cells of the collecting ducts but not the proximal or distal tubules. Binding to the proximal tubules by the lectins Conavalia ensiformis (Con A), Dolichas biflorus, Ricinus communis (RCA-1), and Triticum vulgare and to the distal tubules by Con A, RCA-1, Arachis hypogaea (PNA) with and without neuraminidase, and the antibody for cytokeratins was demonstrated. The lectin binding and cytokeratin staining patterns of rat hereditary renal cell carcinoma, adenoma and the preneoplastic lesions of atypical tubules and hyperplasias suggest that cystic adenomas arise from the distal nephron, principally the collecting duct, whereas the solid atypical tubules, hyperplasias, and adenomas arise from the proximal nephron, principally the proximal tubule.

Human bullous pemphigoid (BP) is an immune-mediated blistering disease characterized by autoantibodies against BP antigens (230/180 kd), which are constitutive glycoproteins of hemidesmosomes found in basal keratinocytes. Blistering diseases similar to human BP have been reported in dogs. IgG deposits at the basement membrane zone (BMZ) are a common feature of canine BP. Although circulating anti-BMZ IgG autoantibodies have been demonstrated in some cases of canine BP, the specific skin protein targeted by these autoantibodies has not been identified. In this study, we characterized the antigenic target of the autoantibodies in the serum from a 3-year-old castrated male Pit Bull Terrier with BP. Direct immunofluorescence of the patient's skin demonstrated IgG deposits at the dermal-epidermal junction. Indirect immunofluorescence demonstrated autoantibodies in the patient's serum that stained the epidermal roof of salt-split canine skin and left the dermal floor unstained. These serum autoantibodies did not stain normal intact dog skin but labeled intact bovine tongue. Direct immunoelectron microscopy of the dog's skin revealed IgG deposits within the hemidesmosomes of the basal keratinocytes. Western immunoblotting experiments showed that canine keratinocytes express both the 230-kd and 180-kd bullous pemphigoid antigens, and the autoantibodies from the patient's serum recognized the 180-kd bullous pemphigoid antigen in proteins extracted from canine and human keratinocytes. Canine BP has many parallel features with human BP including similar immune deposition of IgG within hemidesmosomes and a hemidesmosome-associated 180-kd glycoprotein target for circulating autoantibodies.

Triaryl phosphates, including tricresyl phosphate (TCP) and butylated triphenyl phosphates (BTP), are used in the commercial manufacture of plastics, lubricants, and hydraulic fluids. Recent reports implicate these compounds as endocrine and reproductive toxicants that can cause cholesteryl lipidosis in adrenocortical (AC) and ovarian interstitial (OI) cells, suggesting altered metabolism of steroid hormones or cholesterol or of both. We investigated potential mechanisms of BTP and TCP toxicity to determine if there were functional abnormalities of the adrenal cortex or ovary. Groups of intact (nine or 12) and ovariectomized (six) female F344 rats, 10-12 weeks of age, received 0, 0.4 g/kg TCP, or 1.7 g/kg BTP in sesame oil vehicle or 1.7 g/kg neat BTP for 20, 40, or 60 days. All rats administered BTP and TCP developed cholesteryl lipidosis in AC and OI cells; the TCP-treated group was most severely affected. Serum concentrations of androstenedione and corticosterone were unchanged, but estradiol levels were significantly (P < 0.05) elevated in BTP- and TCP-treated groups (14.5 times and 37.5 times greater than controls, respectively). Vaginal cytology revealed that BTP- but not TCP-treated females had abnormal reproductive cycles that were significantly prolonged in diestrus (3 times greater than control). There were significant elevations in serum total cholesterol (TCP-treated group was 1.3 times greater than controls), low-density lipoprotein (TCP-treated group was 1.8 times greater than controls), alanine transaminase (BTP-treated group was 2 times greater than controls), and albumin (a major serum estradiol-binding protein; BTP-treated group was 4.6 g/dl vs. 3.6 g/dl for controls). Liver weights (134% that of controls) and P-450 enzymes (3 times greater than controls) were significantly increased in BTP-treated rats. Abnormal reproductive cycles, elevated serum albumin, and increased hepatic P-450 concentration suggested fecundity could be affected in female rats exposed to BTP, most likely because of altered liver metabolism. Ovariectomized BTP-treated and control rats had similarly increased uterine weights after challenge with estradiol and estradiol benzoate, indicating that triaryl phosphate-induced esterase inhibition or other xenobiotic-induced block of hormone action in estradiol-responsive tissues was not responsible for the prolonged diestrus in rats with elevated serum estradiol. The pathogenesis of the cholesteryl lipidosis induced by TCP and BTP appeared to be separate from the reproductive effects because the lipidosis was most severe in TCP-treated rats, which had normal reproductive cycles and fertility.

Experimental Marek's disease virus (MDV) infection in chickens was used to study the early pathogenesis of virus-induced atherosclerosis. Previous investigations using this model have reported the occurrence of atherosclerotic lesions after approximately 7 months postinfection. In this study, a total of 75 susceptible Cornell P-line chickens were inoculated intraperitoneally with the CU-2 strain of MDV at 3 days of age and subsequently perfused for histologic examination. At 2, 4, 8, 13, and 20 weeks postinoculation, the ascending aorta and the brachiocephalic and coronary arteries were evaluated for early changes. Expression of class II major histocompatibility complex (Ia) antigen by the vascular endothelium was demonstrated by indirect immunodetection as early as 2 weeks after virus inoculation. This change was followed by significant thickening of the intimai layer associated with mononuclear cell infiltration. All the arteries examined from the MDV-infected chickens were affected. Preliminary immunohistochemical staining showed the presence of CD3+ CD4+, and CD8+ cells among the infiltrating cells. The results suggest that an immunopathologic mechanism may be involved in the early pathogenesis of MDV-induced atherosclerosis in chickens.

A female American black bear (Euarctos ursus americanus) over 20 years old had shown epileptiform neurologic signs starting in March 1992 and was found dead unexpectedly 8 months later. At necropsy, pulmonary and intrabronchial hemorrhage was noted. In the brain, the leptomeninges exhibited slight thickening, and petechiae were evident in the hippocampus. Histopathologic examination of the brain revealed several senile changes: numerous senile plaques, amyloid deposition in cerebromeningeal arterioles, mineral deposition in the pallidum, and numerous corpora amylacea in the cerebellum.

Published surveys of feline neoplasia have not specifically included biliary cystadenoma, and there is only one case report in the literature. This report is a compilation of 13 feline cases and provides a description of clinical, pathologic, immunohistochemical, and ultrastructural aspects of biliary cystadenoma as well as a discussion of comparative pathology of biliary cystadenoma in human beings and speculative histogenesis.

Two of 632 Fischer-344 rats in a food restriction study had spontaneous, bilateral, multicentric renal tubular cell carcinomas. Although there were 104 litters represented in this study, both rats that developed this rare neoplasm were from the same litter. The littermates, one male and one female, were in the food-restricted treatment groups (60% of ad libitum intake) and were 550 and 447 days old, respectively, at death. The probability that the two rare bilateral renal neoplasms occurred by a chance event in littermates is approximately 0.8%. The apparent familial predisposition for development of specific types of neoplasms emphasizes the importance of randomization of individuals into treatment groups and consideration of lineage for rare tumors.

Three male Shar-pei dogs and one female spayed Boxer dog ranging in age from 4 years, 4 months to 7 years, 8 months had 4-7-week histories of diarrhea, progressive anorexia, weight loss, and vomiting. The dogs were depressed, lethargic, and hypoalbuminemic, with losses of 23-39% of body weight. Endoscopic examination revealed gastric ulceration with areas of mucosal hyperemia and pallor. Small nodules with occasional central depressions were present in three dogs. Gastric and proximal duodenal mucosal biopsies showed neoplastic lymphocytes infiltrating the lamina propria, with invasion of the superficial and glandular epithelial basement membrane. Immunohistochemical evaluation of two dogs was negative for anti-canine IgG and positive for anti-CD₃. Necropsy of the Boxer and two Shar-peis confirmed epitheliotropic gastric and small intestinal lymphoma in all three dogs, with mesenteric lymph node and hepatic involvement in two and one dog, respectively.

A 2-year-old spayed female Siamese cat was presented with clinical liver disease characterized by anorexia; depression; elevations in serum levels of alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase; hyperbilirubinemia; and icterus. Liver biopsy diagnosed hepatocellular degeneration with marked centrilobular hepatocellular accumulation of rhodanine-positive brown granules. Subsequent postmortem examination revealed similar granular material in the epithelium of the proximal convoluted tubules and collecting ducts of the kidney and alveolar epithelium and macrophages in the lung. The liver and kidney copper concentrations were 4,074 and 792 ppm dry weight, respectively. Hepatic degeneration in this cat apparently was due to excessive accumulation of copper.

We describe an epithelial neoplasm arising from the fourth ventricle of a goat and extending into the subjacent cerebellum along the spaces of Virchow-Robin. The neoplastic cells are consistent with squamous epithelium based upon light microscopic morphology and cytokeratin immunoreactivity. The lack of overt keratin formation and the lack of an exophytic component distinguishes this neoplasm from intracranial epidermoid cysts, resulting in the classification of this caprine tumor as an epithelioma.

An unusual adenovirus infection in a conventionally raised nursing pig was characterized clinically by hemorrhagic diathesis of the skin and microscopically by disseminated viral inclusions in the endothelial, interstitial, and epithelial cells of the skin, kidney, spleen, liver, heart, and small and large intestines. This disease was confirmed ultrastructurally by demonstration of typical adenoviral particles in the nuclei of endothelial cells in the myocardium and of interstitial cells in the small intestine. This is the first report of disseminated adenovirus infection in a nursing pig in North America.

Five dogs euthanatized because of refractory seizures were found to have hematopoietic elements in the interstitium of the choroid plexus at the level of the fourth ventricle. None of the dogs had significant hematologic or cerebrospinal fluid abnormalities. The extramedullary hematopoiesis was confined to the central nervous system and consisted of megakaryocytes, immature granulocytes, and rubricytes in two dogs and of one predominant cell population in each of the other three dogs. These findings are unique, and factors possibly contributing to the formation of a hematopoietic inductive microenvironment in the choroid plexus are cytokine-neurokine homologies, locally altered vascular supply, and aberrant functioning of bone marrow-derived central nervous system macrophages.

Two of five male Sprague-Dawley rats with hepatic tapeworm cysts developed large multinodular fibrosarcomas. Fibrosarcomas enveloped tapeworm cysts, invaded the serosa of multiple organs, and extended through the diaphragm into the pleural cavity. Light microscopy, immunohistochemistry, and electron microscopy supported the diagnosis of fibrosarcoma. The parasites were identified as Cysticercus fasciolaris, the larval stage of Taenia taeniaeformis. The development of sarcomas in rats induced by Taenia sp. is thought to be attributable to the chronic inflammatory reaction of the capsule. There are parallels between these and other tumors occurring in mice and cats with suggested chronic inflammatory etiologies.