The relation between average duodenal mast cell count, duodenal mucosal mast cell numbers, duodenal connective tissue mast cell numbers, circulating basophil numbers, heterophil-to-lymphocyte ratio, and lesion score were studied to gain an understanding of the events that may lead to intestinal lesion formation associated with hemorrhagic enteritis virus (HEV) infection. Changes in vascular permeability in the duodenum in birds inoculated with HEV were examined, using colloidal carbon and ferritin as vascular markers. Turkeys inoculated with HEV had significantly (P < 0.05) higher duodenal mast cell counts than did noninfected controls. Birds inoculated with HEV had significantly (P < 0.05) more mucosal mast cells than did phosphate-buffered saline solution-inoculated birds. Connective tissue mast cell and basophil numbers were unaffected by viral inoculation. Thermal stress did not have significant effect on lesion severity, but did increase number of birds that developed the characteristic intestinal lesions. The heterophil-to-lymphocyte ratio was significantly (P < 0.05) higher in HEV-inoculated birds, compared with phosphate-buffered saline solution-inoculated controls. Increase in vascular permeability was only detected in HEV-inoculated birds with intestinal lesions. Results indicate that mast cells, and the vasoactive mediators contained within mast cells, may be important in the early manifestation of HEV infection. They also provide a possible mechanism through which biochemical and physiologic changes characteristic of HEV infection can occur.

Immunohistochemistry has been used extensively to evaluate protein expression in clinical and research settings. However, immunohistochemistry is not always successful in veterinary medicine due to the lack of reliable antibody options, poor tissue preservation, labor-intensive staining, and antigen-retrieval optimization processes. RNAscope in-situ hybridization (ISH) is a powerful technology that uses a specific sequence probe to identify targeted mRNA. In this study, we demonstrate RNAscope ISH in 4 common canine malignancies, which are traditionally diagnosed by histopathology and immunohistochemistry. Probes were designed for commonly targeted mRNA markers of neoplastic tumors; these included c-kit in mast cell tumor, microphthalmia-associated transcription factor in malignant melanoma, ionized calcium-binding adapter molecule(-1) in histiocytic sarcoma, and alkaline phosphatase in osteosarcoma. A strong staining signal was obtained by these 4 targets in each canine malignancy. These results support the use of RNAscope ISH for definitive diagnosis in canine malignancies.

OBJECTIVE To assess differences in skin shrinkage between grossly visible tumor and grossly normal marginal skin of dogs for cutaneous mast cell tumors (MCTs) excised with curative intent and to determine an equation to estimate postexcisional gross tumor margins from preexcisional measurements and vice versa. SAMPLE 19 cytologically confirmed and surgically excised cutaneous MCTs obtained from dogs. PROCEDURES Tumors were measured in craniocaudal and dorsoventral directions before excision, immediately after excision, and after fixation in formalin. Both grossly visible tumor and surrounding grossly normal skin that comprised the surgical margin were measured at each time point. Percentage of shrinkage was compared among time points and between the tumor and surrounding grossly normal skin. Patient and histopathologic variables were correlated to skin shrinkage. RESULTS Overall shrinkage was 17.70%. The amount of shrinkage within the grossly visible tumor (4.45%) was less than that within the surrounding grossly normal skin (24.42%). Most of the shrinkage occurred immediately after excision. There was no effect of age, sex, completeness of excision, or degree of edema. Accuracy of an equation to estimate postexcisional margins from preexcisional measurements was only 18.4%. CONCLUSIONS AND CLINICAL RELEVANCE Grossly evident MCTs of dogs shrunk less than did the grossly normal surrounding skin. Although an equation to estimate postexcisional margins from preexcisional measurements could be derived, it likely would need to contain additional variables not included in the study reported here. Until such an equation exists, care must be used when extrapolating surgical margins from histologic margins and vice versa.

Objective—To characterize the response of skin of nonallergic horses following ID injection of polyclonal rabbit anti-canine IgE (anti-IgE) and rabbit IgG. Animals—6 healthy horses. Procedures—Skin in the cervical area was injected ID with anti-IgE and IgG. Wheal measurements and skin biopsy specimens were obtained before and 20 minutes and 6, 24, and 48 hours after injection. Tissue sections were evaluated for inflammatory cells at 4 dermal depths. Immunohistochemical analysis for CD3, CD4, and CD8 was performed, and cell counts were evaluated. Results—Anti-IgE wheals were significantly larger than IgG wheals at 20 minutes and 6 and 24 hours after injection. There were significantly more degranulated mast cells after anti-IgE injection than after IgG injection. There were significantly more eosinophils at 6, 24, and 48 hours and neutrophils at 6 hours after anti-IgE injection, compared with cell numbers at those same times after IgG injection. There were significantly more eosinophils in the deeper dermis of anti-IgE samples, compared with results for IgG samples. No significant differences between treatments were detected for CD3+, CD4+, or CD8+ cells. Conclusions and Clinical Relevance—Injection of anti-IgE antibodies was associated with the development of gross and microscopic inflammation characterized by mast cell degranulation and accumulation of inflammatory cells, particularly eosinophils and neutrophils. This pattern appeared to be similar to that of horses with naturally developing allergic skin disease, although lymphocytes were not increased; thus, ID injection of anti-IgE in horses may be of use for evaluating allergic skin diseases of horses.

Objective-To determine variations in cytologic counts of bronchoalveolar lavage (BAL) fluid attributable to month of collection, first and second aliquots, and left and right lung sites in horses with recurrent airway obstruction (RAO). Animals-5 horses with RAO and 5 healthy horses without respiratory tract disease. Procedures-Horses were housed in a stable for 5 months prior to and throughout the study. Bronchoalveolar lavage fluid was collected from the right and left lung of each horse 3 times at monthly intervals (February, March, and April). Each BAL fluid collection was performed by use of 2 incremental instillations of 250 mL of isotonic saline (0.9% NaCl) solution in the same bronchial site. Analysis of BAL fluid included volume of BAL fluid recovered, a CBC, and differential cytologic counts. Results-Volume of BAL fluid recovered and cytologic counts did not differ in horses with RAO across time or between right and left lungs, except for the number of mast cells. Horses with RAO had significantly lower volumes of BAL fluid recovered, significantly lower percentages of macrophages and lymphocytes, and significantly higher percentages of neutrophils than did healthy horses. Despite individual variation, all horses with RAO had > 25% neutrophils throughout the study period. Conclusions and Clinical Relevance-Despite variation among horses, BAL fluid cytologic counts were repeatable over short and long periods and samples can be used for longitudinal studies as a diagnostic tool of pulmonary inflammation in horses with RAO.

Tracheobronchial aspirates obtained from 66 healthy Thoroughbred racehorses in training at the same track were examined. Twenty-seven percent of the horses had > 20% neutrophils in the aspirate. Eosinophils, mast cells, giant cells, and Curschmann's spirals of mucus were observed in 94, 83, 65, and 42% of the horses, respectively. Hemosiderophages were observed in 86% of the horses, half of which had previous confirmation of exercise-induced pulmonary hemorrhage. Although fungal elements were seen in 70% of the horses, bacteria were detected in only 3% of the horses. The authors conclude that inflammatory airway disease is widespread in the racing Thoroughbred population.