Pauline M. Rakich

Variation among Pathologists in the Histologic Grading of Canine Cutaneous Mast Cell Tumors with Uniform Use of a Single Grading Reference

Ten veterinary pathologists independently assigned histologic grades to the same 60 canine cutaneous mast cell tumors using the Patnaik classifications. The degree of agreement in grading among the pathologists was compared with the degree of agreement among the same pathologists in a previous study, in which each pathologist used the reference for grading that he/she uses routinely. Mean agreement improved significantly from 50.3% to 62.1% with uniform use of the Patnaik classifications (P = 0.00001), suggesting that there is value in uniform application of a single grading scheme for canine cutaneous mast cell tumors. Agreement among pathologists was still not 100%, suggesting that a more objective grading scheme should be developed and that other histologic indicators of prognosis should be investigated.

Variation among pathologists in histologic grading of canine cutaneous mast cell tumors.

Ten veterinary pathologists at 1 veterinary institution independently assigned histologic grades to the same 60 canine cutaneous mast cell tumors (MCTs). There was significant variation among pathologists in grading the MCTs (P < 0.001). The probability of assigning a low grade was significantly higher for the pathologists in this study who use a published reference for histologic grading of canine cutaneous MCTs that allows subcutaneous MCTs or MCTs with mitotic figures to be included in the low-grade category (P < 0.0001 and P < 0.0001, respectively).

ULTRASTRUCTURAL, PROTEIN COMPOSITION, AND ANTIGENIC COMPARISON OF PSITTACINE BEAK AND FEATHER DISEASE VIRUS PURIFIED FROM FOUR GENERA OF PSITTACINE BIRDS

Psittacine beak and feather disease (PBFD) virus, was purified from diseased tissues of a lesser sulphur-crested cockatoo (Cacatua sulphurea), a black palm cockatoo (Probosiger aterrimus), a red-lored Amazon parrot (Amazona autumnalis), and a peach-faced lovebird (Agapornis roseicollis). The histopathology of diseased feathers and follicular epithelium from the different species was compared; basophilic intranuclear inclusion bodies were identified in the follicular epithelium and intracytoplasmic globular inclusions were observed within macrophages located in the feather pulp from the four species. Psittacine beak and feather disease virus antigen was specifically detected by colloidal gold immunoelectron microscopy. The different preparations of purified virions displayed an icosahedral symmetry, were non-enveloped, and had a mean diameter that varied from 12 to 15 nm when negatively stained. Two major viral-associated proteins with approximate molecular weights of 26 and 23 kilodaltons (kd) were consistently demonstrated from the four viral preparations. Purified virions from the four genera were antigenically related. These findings suggest that the PBFD virus purified from numerous genera of diseased birds is similar based on ultrastructural characteristics, protein composition and antigenic reactivity.

Extracutaneous Viral Inclusions in Psittacine Beak and Feather Disease

Thirty-five birds that died with naturally acquired psittacine beak and feather disease (PBFD) were necropsied to identify extracutaneous viral inclusions. Inclusions were found in various tissue sections from 34 of 35 birds. By immunoperoxidase staining, intranuclear and intracytoplasmic inclusion bodies were shown to contain PBFD viral antigen. Inclusion-bearing lesions were widely disseminated but often closely associated with the alimentary tract. Lesions within the palate, esophagus, crop, intestine, bursa of Fabricius, and liver probably serve as sources for viral shedding into the feces.

A Novel DNA Virus Associated with Feather Inclusions in Psittacine Beak and Feather Disease

The nature of feather inclusions was characterized in 32 psittacine birds (30 cockatoos, one peachfaced lovebird (Agapornis roseicollis), and one red-lored Amazon parrot (Amazona autumnalis autumnalis)) with naturally-acquired psittacine beak and feather disease. Intranuclear inclusions within feather epithelial cells and intracytoplasmic inclusions within macrophages in the feather epithelium and pulp cavity contained psittacine beak and feather disease viral antigen when stained by the avidin-biotin complex immunoperoxidase technique. Ultrastructurally, inclusions were observed primarily within macrophages and to a lesser extent within epithelial cell nuclei. Macrophage inclusions appeared as paracrystalline arrays of viral particles. Intranuclear inclusions were less well defined, although scattered viral particles were present. Intracytoplasmic and intranuclear particles in ultrastructural preparations were identified by colloidal gold labeling as psittacine beak and feather disease virus. Feather epithelium was more frequently and severely involved in the disease process than was adjacent follicular epithelium. Plucked feathers with an intact epidermal collar and feather epithelium were preferred to follicular biopsies for histopathologic examination.

Gastrointestinal Pythiosis in Two Cats

Two young adult male Domestic Shorthair cats living in the southeastern United States were evaluated for signs attributable to partial intestinal obstruction. Physical examination indicated a palpable abdominal mass in each animal. Exploratory laparotomy revealed a large extraluminal mass involving the ileum and mesentery with adjacent mesenteric lymphadenopathy in cat No. 1 and an abscessed mass in the distal duodenum in cat No. 2. Mass resection and intestinal anastomosis were performed in both cats. Histologic evaluation indicated that the intestinal lesions involved primarily the outer smooth muscle layer and serosa and consisted of eosinophilic granulomatous inflammation with multifocal areas of necrosis. In Gomori methenamine silver–stained sections, broad (2.5–7.5 μm), occasionally branching, infrequently septate hyphae were observed within areas of necrosis. A diagnosis of Pythium insidiosum infection was confirmed in both cats by immunoblot serology and by immunoperoxidase staining of tissue sections using a P. insidiosum–specific polyclonal antibody. Cat No. 1 was clinically normal for 4 months after surgery but then died unexpectedly from an unknown cause. Cat No. 2 has been clinically normal for at least 9 months after surgery and appears to be cured on the basis of follow-up enzyme-linked immunosorbent assay serology.

Effects of cyclosporin A administration in cats.

Cyclosporin A was administered orally to 10 cats for 28 consecutive days at a dosage of 20 mg/kg body weight daily divided into 2 equal doses. Serum trough CyA concentrations ranged from 134 to 902 ng/ml with a mean of 567 +/- 249 ng/ml (means +/- SD). Immunosuppression by CyA was suggested in 5 cats by significantly depressed lymphoblast transformation responses. Various hematologic, serum chemical, and urinalysis parameters were monitored on a weekly basis in 10 cats. Serum urea nitrogen concentration increased significantly from baseline values on days 7, 14, and 21, but not on day 28. Urine concentrating ability was unimpaired. Alanine aminotransferase activity was decreased significantly from baseline values at each sample period during CyA administration. Drug-related side effects were minor; one cat developed gingival hypertrophy which regressed within 21 days of CyA withdrawal.

RESPONSES OF RED FOXES TO FIRST AND SECOND INFECTION WITHSARCOPTES SCABIEI

The clinical response of red foxes (Vulpes vulpes) to the mange mite, Sarcoptes scabiei, was characterized by infection of five, 4-mo-old red foxes with S. scabieioriginally isolated from a wild red fox. The infected foxes and three uninfected control foxes were monitored with weekly complete blood counts and biweekly serum chemistry profiles, hypersensitivity tests, and evaluation of skin biopsies. After 7 wk, the foxes were treated and held free of infection for 2 mo. Six foxes, three previously infected and three with no history of exposure, were then infected with the same isolate of S. scabiei and followed for another 7 wk; two additional previously infected foxes were held as treatment controls, and two foxes with no history of exposure as naive controls. All infected foxes developed significant immediate (Type I) hypersensitivity reactions to a S. scabiei mite extract within 2 wk of exposure and maintained this reaction as long as 4 mo after clearance of mites. Pronounced mast cell hyperplasia and infiltration with eosinophils were the earliest inflammatory cell responses noted in biopsy samples from infected foxes and were maintained throughout infection. Infected foxes also showed significant increases in white blood cell counts, due primarily to increases in numbers of circulating neutrophils and eosinophils. Clinical response, severity of disease, and relative numbers of mites per cm2 of skin of previously infected foxes and foxes undergoing their first infection did not differ. These results show that red foxes develop strong immediate hypersensitivity reactions to S. scabieibut, under our experimental conditions, did not exhibit resistance to reinfection.

Genetic Variability of Cytauxzoon Felis from 88 Infected Domestic Cats in Arkansas and Georgia

Although cytauxzoonosis has historically been nearly 100% fatal in domestic cats, increasing number of reports of infected cats that demonstrate less-severe disease suggest the existence of different strains of Cytauxzoon felis. To test this hypothesis, the genetic variability of C. felis was examined in blood samples from naturally infected domestic cats from Arkansas and Georgia by using the first and second ribosomal internal transcribed spacer regions (ITS1, ITS2) as markers to assess genotypic variability. In addition, the clinical outcome of infection (survival vs. fatal disease) was analyzed. Within the C. felis ITS1 region, there were a total of 8 single nucleotide polymorphisms (SNP) and a single nucleotide insertion. Within the ITS2 region, there were a total of 4 SNPs and a single 40 base pair insertion. When taken together, the ITS1 and ITS2 sequence data defined a total of 11 different sequences and 3 unique genotypes. One unique ITS1-ITS2 genotype was detected in samples submitted exclusively from Arkansas, and a second unique genotype was submitted exclusively from Georgia. There was a significant association between infection with C. felis that contained particular ITS genotypes and survival of the infected domestic cat. The identification of unique C. felis genotypes obtained from different geographic areas and the association of particular ITS genotypes with the outcome of infection suggest the existence of parasite strains that may vary in pathogenicity to the domestic cat and offer an explanation for the survival of some infected cats in more recent case studies.

Acute hepatic sarcocystosis in a chinchilla

Formalin-fixed sections of heart, liver, lung, kidney, spleen, and intestine from a 7-month-old female pet chinchilla were submitted for histologic examination to the Athens Diagnostic Laboratory, University of Georgia. The animal had been purchased at 4 months of age and was 1 of a litter of 2; its littermate had died within hours of birth from an unknown cause. At the time of death, the subject of this report was housed with 6 related chinchillas in a single large cage and was fed chinchilla and guinea pig feeds, dried and fresh vegetables, rose hips, alfalfa, and raisins. No abnormalities were seen in any of the animals until the chinchilla of this report was found dead, after last being observed as normal less than 2 hours previously. Paraffin-embedded sections of tissues were cut at 3 μm and stained with hematoxylin and eosin (HE) or periodic acidSchiff (PAS). Paraffin-embedded sections were dewaxed and reacted with antisera to Toxoplasma gondii, Neospora caninum, and Sarcocystis cruzi by an avidin-biotin complex (ABC) immunohistochemical staining procedure. Microscopically, the liver was characterized by marked disruption of architecture by numerous randomly distributed coalescing areas of inflammation and necrosis (Figs. 1, 2). Inflammatory infiltrates consisted primarily of neutrophils and macrophages. Sinusoids throughout the liver were filled with large blast-type cells interpreted as representing immature blood cells (extramedullary hematopoiesis). Various stages of protozoa were scattered within the areas of necrosis and inflammation (Figs. 3-8). The lung was congested and atelectatic. Septa were thickened by fibrin and increased numbers of mixed inflammatory cells. Alveoli contained macrophages and proteinic fluid. No parasites were evident, and no bacteria or fungi were detected with Brown and Brenn, Giemsa, and Gomori’s methenamine silver stains. Blood vessels in all tissues contained large numbers of immature white blood cells, but no inflammation or evidence of parasites was seen in any other tissues. Protozoa1 schizonts in the liver divided by endopolygeny, a process in which the parasite nucleus becomes multilobed before formation of numerous merozoites (Figs. 4-8). The earliest schizont (9 x 7 μm) contained 1 nucleus with a prominent nucleolus (Fig. 3). The nucleus then enlarged and became multilobed (Figs. 4, 5). Schizonts varied in shape and the number of nuclei. Merozoites budded peripherally, often forming a rosette around a central residual body (Fig. 7). The residual body was eosinophilic, diffuse, and up to 7 μm in diameter. Schizonts with merozoites were up to 25 x