Cytologic Criteria for Mast Cell Tumor Grading in Dogs With Evaluation of Clinical Outcome.

A 2-tiered histologic grading scheme for canine cutaneous mast cell tumors (MCTs) is based on morphologic characteristics of neoplastic cells, including karyomegaly, multinucleation, nuclear pleomorphism, and mitotic figures. Aspirates from MCTs may provide the same information more quickly, inexpensively, and less invasively. This study used these criteria to develop a cytologic grading scheme for canine MCTs to predict outcome. Three anatomic pathologists graded histologic samples from 152 canine MCTs. Three clinical pathologists evaluated aspirates from these masses using similar criteria. A cytologic grading scheme was created based on correlation with histologic grade and evaluated with a kappa statistic. Survival was evaluated with Kaplan-Meier survival curves. Cox proportional hazards regression was used to estimate hazard ratios for tumor grades and individual grading components. Simple logistic regression tested for relationships between risk factors and mortality. The cytologic grading scheme that best correlated with histology (kappa = 0.725 ± 0.085) classified a tumor as high grade if it was poorly granulated or had at least 2 of 4 findings: mitotic figures, binucleated or multinucleated cells, nuclear pleomorphism, or >50% anisokaryosis. The cytologic grading scheme had 88% sensitivity and 94% specificity relative to histologic grading. Dogs with histologic and cytologic high grade MCTs were 39 times and 25 times more likely to die within the 2-year follow-up period, respectively, than dogs with low grade MCTs. High tumor grade was associated with increased probability of additional tumors or tumor regrowth. This study concluded that cytologic grade is a useful predictor for treatment planning and prognostication.

Comparison of CO(2) laser and 4.0 MHz radiosurgery for making incisions in the skin and muscles of green iguanas (Iguana iguana).

Four green iguanas scheduled for euthanasia were used to compare the extent of collateral tissue damage associated with CO(2) laser and 4.0 MHz radiosurgery. The iguanas were anaesthetised and a series of three skin and three muscle incisions was made by 4.0 MHz radiosurgery (0.18 mm wire electrode, 25 W, cut mode) and CO(2) laser (0.3 mm ceramic tip, 15 W focused beam super-pulse mode), and three incisions were made with a scalpel blade as controls. Following euthanasia, a total of 60 skin and 36 muscle sections were evaluated histologically. Radiosurgery and the laser both produced bloodless incisions, but radiosurgery caused significantly less collateral tissue damage in the skin (307 [97] v 386 [108] microm) and the muscle (18 [7] v 91 [15] microm).

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.

Responses of red foxes to first and second infection with Sarcoptes 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. scabiei originally 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. scabiei but, under our experimental conditions, did not exhibit resistance to reinfection.

Diseases diagnosed in red foxes from the southeastern United States.

Diagnostic findings on 51 red foxes (Vulpes vulpes) from the southeastern United States examined from 1967 to 1995 were reviewed. Etiologic diagnoses included sarcoptic manage (n = 33), traumatic injury and associated complications (n = 5), suspected canine distemper (n = 2), capture myopathy (n = 1), congenital absence of guard hairs (n = 1), intradermal tick infestation (n = 1), otodectic manage (n = 1), and toxicosis (n = 1). The cause of morbidity was not determined for three of the foxes, and three others were classified as normal animals. Sarcoptic manage was diagnosed in 65% of the red foxes, was found in foxes submitted from four of the eight southeastern states represented, and was seen in 19 of 29 yr covered by this study.

Trombidiosis-induced dermatitis in white-tailed deer (Odocoileus virginianus).

During a routine blood collection at a wildlife management checkpoint, several white-tailed deer (Odocoileus virginianus) from Lake Russell Wildlife Management Area, Habersham County, Georgia, were observed to have accumulations of orange granular material around the eyes and mouth accompanied by serous exudate, crusting, and alopecia. Microscopic examination of the granular material disclosed numerous larval mites of the family Trombiculidae. Histologic examination revealed severe chronic diffuse plasmacytic dermatitis with intralesional larval trombiculid mites. Several stylostomes were also identified in the tissue sections. This paper describes the gross and histologic changes associated with severe trombiculid infestation in a white-tailed deer and suggests potential causes for this unusual finding.

Treatment of localized Mycobacterium avium complex infection with clofazimine and doxycycline in a cat.

Mycobacterium avium complex infection resulted in a granuloma that developed at the base of the left ear in a cat. The lesion caused vestibular dysfunction and facial palsy on the left side and protruded into the oral cavity on that side. The cat was treated successfully, with resolution of the lesion and elimination of the organism, by use of combined administration of clofazimine and doxycycline. Adverse effects of the clofazimine treatment included temporary reddish-orange discoloration of the cat's skin and mucous membranes.

Rapid tissue expansion for the development of rotational skin flaps in the distal portion of the hindlimb of dogs: an experimental study.

In phase I, tissue expanders were implanted subcutaneously over the lateral crural region of four dogs. The expanders were inflated daily to maintain a constant intraluminal pressure for 7 days. All animals tolerated the pressurized expanders well. Some animal patient discomfort was associated with repeated injections into the subcutaneous filling port. One dog developed a limited area of tissue loss and exposure of the expander. Tissue response to the expanders included epidermal hyperplasia, dermal collagen compression with early fibroplasia, and subcutaneous granulation tissue formation adjacent to the expander pocket. In phase II, a second group of four dogs underwent rapid skin expansion over 7 days. The pressure applied to the expanding skin was determined daily and adjusted to approximate the value reported for capillary pressure in the dog. Tissue discoloration occurred in two dogs, suggestive of impaired circulation. Skin flaps were developed from expanded skin and rotated over the talocrural region. Wound dehiscence occurred along the distal flap margin in three phase II dogs. These wounds healed by second intention. Branches of the caudal saphenous artery were identified, using angiography, as the primary blood supply to the skin flaps.

Clinical and histologic characterization of cutaneous reactions to stings of the imported fire ant (Solenopsis invicta) in dogs.

Four adult dogs received experimentally controlled stings in the dorsolateral abdominal skin by imported fire ants (Solenopsis invicta). The sites were examined grossly 15 minutes and at 1, 2, 3, 4, 5, 6, 24, 48, and 72 hours and histologically 15 minutes and 6, 24, 48, and 72 hours after stinging. The initial gross lesions at 15 minutes were swelling and erythema, and the microscopic changes were vascular congestion and superficial dermal edema. By 6 hours, the lesions consisted of bright erythematous pruritic papules characterized microscopically by a band of full thickness dermal necrosis and inflammation. By 24 hours and continuing to the end of the study at 72 hours, the sites appeared completely normal grossly. Biopsies taken 24, 48, and 72 hours after stings contained microscopic changes similar to those present at 6 hours after stings. These histologic changes are unlike those described for human beings stung by imported fire ants. In human beings, fire ant stings are characterized histologically by an initial superficial vesicle that evolves into a sterile pustule.

Cryptosporidiosis in four cockatoos with psittacine beak and feather disease.

Cryptosporidiosis was diagnosed in 4 cockatoos with psittacine beak and feather disease. Three of the birds had cryptosporidiosis confined to the epithelium covering the bursa of Fabricius. One bird had generalized parasitism of the small intestine, large intestine, and bursal epithelium. All of the birds had intermittent to protracted diarrhea before death. Presumably, acquired immunodeficiency from psittacine beak and feather disease promoted establishment of cryptosporidiosis and other secondary diseases including septicemia, peritonitis, chlamydiosis, and mycotic ventriculitis.

Comparison of procedures for assessing adequacy of dog foods.

Dog foods with similar claims for nutritional adequacy were tested by chemical analysis and the American Association of Feed Control Officials' growth trial. All foods tested were similar chemically, however, dogs given one regionally marketed food had lower growth rate and food efficiency as well as suboptimal PCV and hemoglobin values during the growth trial. Pups fed this diet also had clinical signs typical of zinc and copper deficiencies. We concluded that American Association of Feed Control Officials' approved feeding tests provide valid assessment of pet food quality, and procedures involving only chemical analysis or calculated values may not.

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 peach-faced 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.

Evaluation of ketoconazole and itraconazole for treatment of disseminated cryptococcosis in cats.

During the first part of a study, cats were inoculated with Cryptococcus neoformans via the following routes: intradermal, intranasal, IV, and intracisternal. Only use of the IV route of inoculation consistently induced disseminated cryptococcosis. In the second part of the study, disseminated cryptococcosis was experimentally induced in cats via IV inoculation of C neoformans. One month after inoculation, 3 cats were treated with ketoconazole (10 mg/kg of body weight/d) and 3 cats were treated with itraconazole (10 mg/kg/d) for 3 months. One of the ketoconazole-treated and 2 of the itraconazole-treated cats also had cryptococcosis of the CNS when treatment was begun. During treatment, serum cryptococcal antigen titer progressively decreased in all cats. Abnormalities in CBC values or the serum biochemical profile were not found in any cat during treatment. However, all ketoconazole-treated cats became anorectic and lost weight. Side effects were not seen in itraconazole-treated cats. During the 3-month posttreatment observation period, all cats remained healthy. At necropsy, histologic evidence of cryptococcosis was not found in the 3 ketoconazole-treated cats or in 2 of the itraconazole-treated cats. In the third itraconazole-treated cat, cryptococcal organisms were found in the kidneys.

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.

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.

Rabies-like inclusions in dogs.

Inclusion bodies, indistinguishable from rabies inclusion bodies (Negri bodies), were found in the brains of 8 nonrabid dogs. The inclusions were compared to Negri bodies present in neurons of rabies-positive animals and examined for the presence of rabies virus by a combination of immunoperoxidase staining (7 cases), fluorescent antibody (FA) staining (1 case), and transmission electron microscopy (4 cases). Positive immunoperoxidase staining for rabies was obtained in brain tissues from FA rabies-positive animals. All brain tissues from the 7 dogs stained by the immunoperoxidase method and the brain from the 1 dog stained by the FA method were negative for rabies. Rabies virus was not found in inclusion-containing neurons in the cases examined by transmission electron microscopy. These results emphasize the importance of FA testing and mouse inoculation for the diagnosis of rabies.

Cytology of the respiratory tract.

Information has been presented to assist veterinary practitioners in obtaining and interpreting cytology specimens from the respiratory tract. The various cytologic techniques are rapid, economical, and relatively noninvasive. Cytologic examination of the respiratory tract may provide a definitive diagnosis or information for improved case management until more complex diagnostic procedures are performed.

Clinical interpretation of leukocyte responses.

Basic information has been presented concerning leukocyte (neutrophil, monocyte, lymphocyte, eosinophil, and basophil) function, production, kinetics, and response to various physiological and disease states. Using this information, veterinary practitioners should be able to interpret leukogram data from sick and healthy dogs and cats. Specifically, characteristic leukogram patterns such as physiological leukocytosis, corticosteroid-associated changes, and the presence of infection or severe inflammation should be recognized. In addition, interpretation of individual leukocyte responses should be possible. Several tables have been provided to assist in constructing a differential diagnosis to explain increases or decreases in absolute leukocyte numbers that lie outside of expected reference intervals.