Mouse hepatoblastomas: a histologic, ultrastructural, and immunohistochemical study

Hepatoblastomas from B6C3F1 and BALB/c mice were examined by light and electron microscopy and by immunohistochemical reactions for alpha-fetoprotein, keratin, and vimentin. Tumors occurred in one group of a chronic bioassay for the interaction of diet, genetic strain, and the carcinogen, 2-acetylaminofluorene. Tumors had several populations (including epithelial and mesenchymal cells) in various stages of differentiation. Neoplastic epithelial cells had features of embryonal hepatocytes, such as sparse cytoplasmic organelles, absence of glycogen, abundant free ribosomes, occasional bile canaliculi, and peroxisome-like dense bodies. Embryonal fibroblast-like cells had pleomorphic and folded nuclei with prominent perinuclear chromatin and dispersed cytoplasmic organelles. Fibroblast-like cells were surrounded by bundles of collagen fibrils. Intermediate or transitional types of cells were seen. No tumor cells were immunoreactive for mouse alpha-fetoprotein (AFP) antibody, unlike those in hepatocellular adenomas or carcinomas. Epithelial and mesenchymal tumor cells contained intermediate filaments throughout the cytoplasm; some of these cells stained for keratin but not for vimentin. These findings suggest that mouse hepatoblastomas are derived from bipotential liver blastema cells and are composed of a mixture of several cell populations.     

Mammary carcinoma in a tiger (Panthera tigris): morphological and immunohistochemical study

The histologic and immunohistochemical features of a case of mammary gland carcinoma are described in a 14-yr-old female tiger (Panthera tigris). Immunoreactivity to estrogen receptor (ER), progesterone receptor (PR), tumoral protein 53 (p53), vascular endothelial growth factor (VEGF), human epidermal growth factor receptor 2 (HER-2), and cyclooxigenase-2 (COX-2) was investigated. Neoplastic cells were negative for ER, PR, and p53 but showed positivity for VEGF, HER-2, and COX-2, both in the primary and the metastatic lesions. Histopathologic findings and immunohistochemistry results suggested that the malignant behavior of the reported case could be comparable with some aggressive cat mammary carcinomas.     

Cell proliferation in feline normal, hyperplastic and neoplastic mammary tissue--an immunohistochemical study

The expression of a proliferation cell marker in neoplastic and non-neoplastic mammary tissue in 31 cats was assessed by immunohistochemistry in formalin-fixed paraffin-embedded samples using a monoclonal antibody against nuclear antigen Ki-67 (MIB-1). The results revealed that cell proliferation was more intense in hyperplastic than in normal mammary tissue. Carcinomas exhibited a higher MIB-1 index than benign tumours. There was also a positive correlation between proliferative activity and the histological grade of carcinomas. Fibroadenomatous change, which is considered to be hyperplastic and associated with favourable biological behaviour, exhibited high proliferative activity involving both epithelial and mesenchymal components. MIB-1 detected in formalin-fixed material with pre-treatment with antigen retrieval solution appeared to be a reliable marker of proliferation in feline mammary tumours, but further studies are needed to investigate the value of this proliferation marker in predicting clinical outcome and/or as a prognostic factor in feline mammary lesions.     

Fibrosarcoma with typical features of postinjection sarcoma at site of microchip implant in a dog: histologic and immunohistochemical study

A 9-year-old, male French Bulldog was examined for a subcutaneous mass located at the site of a microchip implant. Cytologic examination of the mass was suggestive of a malignant mesenchymal neoplasm. Histologically, the mass was confirmed as a high-grade infiltrative fibrosarcoma, with multifocal necrosis and peripheral lymphoid aggregates. By immunohistochemistry, the sample was investigated for vimentin, smooth-muscle actin (SMA), CD3, CD79alpha, and CD18. All the neoplastic cells were positive for vimentin. Scattered cells at the periphery of the lesion were also positive for SMA, highlighting a myofibroblastic phenotype. The lymphoid cells were positive for CD18 and CD3. No aluminum deposits were detected by the aurintricarboxylic acid method. A diagnosis of fibrosarcoma morphologically similar to feline postinjection sarcomas was made. Fibrosarcomas at the site of injections have been reported in dogs and ferrets. Furthermore, neoplastic growth at the site of microchip implant in dog and laboratory rodents has been described.     

Intestinal candidiasis in a loggerhead sea turtle (Caretta caretta): an immunohistochemical study

Post mortem examination of a juvenile loggerhead sea turtle (Caretta caretta) stranded in the Canary Islands revealed a fishing-line in the small intestine. Histologically, severe necrotic enteritis, multiple haemorrhages, and marked oedema of the intestinal submucosa were observed. Yeast cells and fungal hyphae were seen in the lamina propria of the intestinal mucosa and in the connective tissue of the submucosa. Because fungal cultures were not taken at the time of necropsy, an immunohistochemical study was performed in order to identify the fungus involved. Specific monoclonal and heterologously absorbed polyclonal antibodies served as the primary reagents for identification of aspergillosis, candidiasis, fusariosis, geotricosis, scedosporiosis, and zygomycosis, using an indirect immunofluorescence staining technique. The fungal elements were strongly stained only by a polyclonal antibody against Candida albicans and a monoclonal antibody against C. albicans. There are no known previous reports of Candida sp. causing skin disease or systemic mycotic infection in sea turtles.     

Pleural endometriosis in an aged rhesus macaque (Macaca mulatta): a histopathologic and immunohistochemical study

Endometriosis is defined as the presence of endometrial tissue outside the uterine cavity and is one of the most common reproductive abnormalities encountered in women as well as Old World primates. The majority of endometriosis cases in Old World primates occur within the abdominal cavity, with spread to extraabdominal sites considered to be a rare event. A 19-year-old multiparous female rhesus macaque (Macaca mulatta) presented to necropsy for difficulty breathing and weight loss. Grossly, the animal had marked abdominal endometriosis and severe hemoabdomen and hemothorax, the latter of which was accompanied by marked pleural fibrosis. Histologic examination confirmed the abdominal endometriosis and also revealed numerous uterine glands and stroma embedded within the pleural fibrosis. Rafts of endometrial tissue were present within pulmonary lymphatics and the tracheobronchial lymph nodes. Immunohistochemically, all ectopic endometrial tissue had varying degrees of positive immunoreactivity to cytokeratin, vimentin, progesterone and estrogen receptors, and calretinin but was negative for desmin and carcinoembryonic antigen. Pleural endometriosis is an extremely rare manifestation of endometriosis in nonhuman primates. This case report emphasizes lymphatic spread as a likely mechanism for extrauterine endometriosis.     

Paratuberculosis in red deer (Cervus elaphus): an immunohistochemical study

In the present study, we compared the utility of immunohistochemistry with serological and histological results for the characterization of Mycobacterium avium ssp. paratuberculosis (M. paratuberculosis) in tissues of affected red deer. Bacterial isolation was considered the standard reference. Samples were taken from seven clinically affected animals with typical macroscopic lesions. The enzyme linked immunosorbent assay (ELISA) and the gel diffusion tests (GD) were used for serological determinations. Samples from intestine and mesenteric lymph nodes were processed for bacterial isolation and histology. M. paratuberculosis was isolated from all the animals. Histologically, lymph nodes displayed necrosis and mineralization at the cortical and medullar areas. Ziehl-Neelsen stained bacteria were numerous inside macrophages and Langhans-type giant cells. Giant and epithelioid cells and lymphocytes were prominent at the ileal mucous membrane. The immunostaining of M. paratuberculosis was very clear inside epithelioid and giant cells. Image analysis was carried out to determine the immunostained area. There was total agreement among the methods employed. Immunohistochemistry can be very useful when the microorganism cannot be recovered from tissues or faeces.     

A sequential histologic and immunohistochemical study of duck hepatitis B virus infection in Pekin ducks

Twenty-nine Pekin ducks were inoculated with duck hepatitis B virus (DHBV), DHBV-free serum, or saline at 1 day of age. Congenitally DHBV-infected ducks were also studied. Ducks were killed periodically during a 92-week study and examined histologically and immunohistochemically to assess liver and extrahepatic inflammation and to detect and characterize DHBV core antigen tissue distribution. DHBV infection produced an asymptomatic but persistent DHBV viremia in all ducks associated with a mild to moderate transient hepatic inflammation apparent at 3 to 6 weeks post-inoculation and waning afterwards. DHBV core antigen was detected in hepatocyte cytoplasm at 1 week post-inoculation, and by 3 weeks post-inoculation scattered pancreatic acinar and islet cells also contained viral antigen. Small numbers of mononuclear cells in the splenic white pulp also contained viral antigen. Viral antigen persisted in all of these tissues throughout the duration of the experiment. No inflammation or tissue injury was detected in any of the extrahepatic tissues during the course of DHBV infection. One DHBV-injected duck developed a hepatocellular carcinoma at 88 weeks of age. Isolated patches of neoplastic hepatocytes contained cytoplasmic DHBV core antigen. The results of this study indicate that DHBV, like mammalian hepadnavirus, is capable of producing a persistent infection of the liver and several extrahepatic tissues and suggest that persistent infection may be associated with the development of hepatocellular carcinoma.     

Cytologic, histologic, and immunohistochemical features of lingual liposarcoma in a dog

A 9-year-old female spayed mixed-breed dog was presented to the referring veterinarian with a history of decreased appetite and difficulty with prehension and swallowing because of a firm oval mass in the tongue. On cytologic evaluation of a fine-needle aspirate of the mass there were numerous round to polygonal cells organized individually or in loose clusters with rare branching capillaries. The cells had eosinophilic granular cytoplasm, round to oval nuclei, and occasionally indistinct borders. The cytologic diagnosis was granular cell tumor (GCT) of the tongue. Impression smears of a biopsy sample of the lingual mass contained similar eosinophilic granular cells with variable numbers of clear vacuoles in the background, numerous perivascular arrangements, and occasional lipoblasts, suggestive of liposarcoma. On histologic examination the tumor was composed of numerous lipocytes with rare foci of round eosinophilic granular cells without evidence of vacuolation; occasionally, atypical mitotic figures were seen. Immunohistochemically, the cells were uniformly negative for periodic acid-Schiff and did not express smooth muscle actin, desmin, or cytokeratin but were immunoreactive for vimentin and S100. A diagnosis of well-differentiated liposarcoma was made on the basis of morphologic and immunohistochemical results. Eosinophilic granular cells may be a component of well-differentiated liposarcoma and are not limited to GCT. Liposarcoma should be considered in the differential diagnoses of lingual tumors in the dog when cytological evaluation reveals eosinophilic granular cells consistent with GCT.     

Reovirus infection in psittacine birds (Psittacus erithacus): morphologic and immunohistochemical study

In this paper we report on an outbreak of reovirus, herpesvirus (Pacheco disease), and/or mycosis infection (Aspergillus spp. and Zygomyces spp.) affecting a batch of young African grey parrots (Psittacus erithacus), with 80% morbidity and 30% mortality. Study material was taken from five birds (four dead and one euthanatized) with a range of clinical symptoms (depression, diarrhea, respiratory symptoms). Diagnosis was confirmed by immunohistochemical detection of avian reovirus, electron microscopy, and virus isolation. Viral antigen of reovirus was detected mainly in large mononuclear cells in the bursa of Fabricius and the spleen, pancreas epithelial cells, and circulating cells; lymphoid organs displayed the largest number of immunopositive cells and severe lymphocyte depletion. Bacteriologic study was negative. Reovirus infection was common in all birds studied, whereas Pacheco disease and mycosis were found in only some, suggesting that reovirus could be the initial cause triggering the outbreak and facilitating infection by other agents and their swift spread through the batch.     

Spastic syndrome in a Holstein bull: a histologic study

A 4-year-old Canadian holstein bull developed the spastic syndrome, an episodic but progressive disorder causing pelvic limb muscular spasms. A post-mortem study, including morphometry of skeletal muscles and teased peripheral nerve fibers of the pelvic limb, revealed mild type II skeletal muscle fiber atrophy and minimal, focal segmental demyelination with remyelination, and axonal degeneration in peripheral nerves. Such alterations are probably incidental or age-associated. Idiopathic muscular cramps is the most probable explanation of the clinical disease and is consistent with the absence of significant morphologic pathologic lesions.     

An immunohistochemical study of interstitial cells of Cajal (ICC) in the equine gastrointestinal tract.

The interstitial cells of Cajal (ICC) are c-kit immunoreactive cells of the gastrointestinal tract which are suggested to have a role in the control of intestinal motility. Cells with c-kit immunoreactivity have not been previously described in the gastrointestinal tract of the horse. Immunoreactivity for c-kit was revealed using immunohistochemical labelling with an anti-c-kit polyclonal antibody. Sections of normal gastrointestinal tissue were examined from 13 anatomically defined sites from stomach to small colon taken from horses free from gastrointestinal disease. Three types of c-kit immunoreactive cells were identified: spindle-shaped cells in the region of the myenteric plexus, stellate or bipolar cells in the circular muscle layer, and round cells in the submucosa. The round cells were shown to be mast cells with the use of toluidine blue staining, whereas the other c-kit immunoreactive cells did not exhibit metachromasia and were classified as ICC. This study will serve as a basis for future pathological studies in the horse.     

Cutaneous plasmacytomas in dogs: a morphologic and immunohistochemical study

Forty-nine cutaneous plasmacytomas in 46 dogs were studied. Tumors occurred at solitary sites in middle-aged to old dogs (mean age, 9.7 years) and most commonly involved the skin of the digits, lips, and ears. Initial diagnosis was made on the basis of light microscopic morphologic findings. Tumors were graded according to the extent of cellular differentiation and immunoreactivity to a panel of immunohistochemical markers (cytokeratins, canine IgG F[ab]2, neurofilament, neuron-specific enolase, S-100 protein, and vimentin). Immunoreactivity was limited to antibodies directed at canine IgG F(ab)2 and vimentin. Vimentin immunoreactivity was usually greater than that of canine IgG F(ab)2, but there was no correlation between immunoreactivity and histologic grade of the tumors. Thirty-six of 39 dogs (92.3%) followed (mean follow-up, 13 months) were cured by surgical excision. The results of this study indicate that canine cutaneous plasmacytomas are benign neoplasms that should be included in the differential diagnosis of cutaneous round cell tumors in dogs.     

Cytologic,histologic, and immunohistochemical features of maxillofacial alveolar rhabdomyosarcoma in a juvenile dog

Abstract: A 15‐month‐old castrated male dog with a history of intermittent epistaxis and sneezing was admitted for the examination of a maxillofacial mass. An impression smear of a biopsy sample from the cauliflower‐shaped gingival mass contained numerous round cells, 5–25 μm in diameter, which contained a moderate amount of clear to pale blue cytoplasm and resembled lymphoid cells. Mitotic figures were frequently observed. The mass was diagnosed as malignant round cell neoplasia. On histologic examination the tumor was composed of diffusely arranged, small, atypical round cells with a small amount of fibrovascular stroma. Immunohistochemically, the cells were negative for CD3, CD18, CD20, CD79α, cytokeratin, melan‐A, chromogranin A, α‐smooth muscle actin, and myoglobin but positive for vimentin and desmin. The cells also had strong positive nuclear staining for myogenin and MyoD1. A diagnosis of solid‐pattern alveolar rhabdomyosarcoma was made on the basis of morphologic and immunohistochemical results. Alveolar rhabdomyosarcoma should be considered in the differential diagnosis of tumors in juvenile dogs, especially when cytologic findings reveal round, undifferentiated cells.     

Application of diethylstilbestrol dipropionate in bulls. I. Excretion of residues in urine and faeces and histological and immunohistochemical changes in the prostate

In this experiment 20 one year old bulls received a single intramuscular injection of the anabolic preparation diethylstilbestrol dipropionate (DES-DP) (an oil preparation or an emulsion). Four animals received a corresponding placebo. The application of DES-DP to bulls caused characteristic histological alterations in the peripheral glandular epithelium of the prostate, which could be observed until four weeks after treatment. The value of histological investigation as a screening method was, however, limited by the occurrence of only few metaplastic lesions and a rapid recovery. By contrast, immunohistochemistry using a polyclonal cytokeratin antiserum K40 appeared to be a specific and very sensitive method to detect oestrogen-induced lesions in the prostate. In only two animals, six weeks after injection with the DES-emulsion, false-negative results were obtained, demonstrating the potential value of this screening method. The excretion of DES in the urine and faeces was monitored using radioimmunoassay following chromatographic purification of the urine and faeces extracts. The excretion of DES in urine was faster for animals of the oil group. The DES content in urine decreased to the 1 microgram/l level after 42 days (emulsion group) or 70 days (oil group). The excretion in faeces was comparable to that in urine. After day 21 the excretion patterns of the two excreta were indistinguishable.     

An immunohistochemical study of the gastrointestinal endocrine cells in the C57BL/6 mice

The regional distributions and relative frequencies of some gastrointestinal endocrine cells in the eight portions (fundus, pylorus, duodenum, jejunum, ileum, cecum, colon and rectum) of the gastrointestinal tract of C57BL/6 mouse was studied with immunohistochemical method using seven types of specific anti-sera against chromogranin A (CGA), serotonin, somatostatin, human pancreatic polypeptide (HPP), glucagon, gastrin and cholecystokinin (CCK)-8. In this study, all these seven types of immunoreactive (IR) cells were identified. Most of these IR cells in the intestinal portion were generally spherical or spindle in shape (open-type cell) while cells showing round in shape (closed-type cell) were found in the intestinal gland and stomach regions occasionally. Their relative frequencies were varied according to each portion of gastrointestinal tract. CGA-IR cells were demonstrated throughout the whole gastrointestinal tract and they showed most predominant in the pylorus and duodenum. Serotonin-IR cells were detected throughout whole gastrointestinal tract and they showed highest frequency in the stomach and colon. Somatostatin-IR cells were demonstrated throughout whole gastrointestinal tract except for large intestine and showed highest frequency in the fundus. HPP-IR cells were found in the fundus with rare frequency. Peculiarly, glucagon-IR cells were restricted to the fundus, ileum and colon with a few frequencies. Gastrin-IR cells were restricted to the pylorus with numerous frequency and CCK-8-IR cells were observed in the pylorus, duodenum and jejunum with numerous and/or a few frequencies, respectively. In conclusion, some peculiar distributional patterns of gastrointestinal endocrine cells were found in C57BL/6 mouse.     

Hyperplastic goitre and mortality in captive-reared black stilts ( Himantopus novaezelandiae )

AIM: To study the gross, histopathological and clinico-pathological findings in cases of hyperplastic goitre in sub-adult captive- reared black stilts following their release on riverbeds in the south Canterbury region of New Zealand. METHODS: Necropsies were undertaken on the recovered carcasses of 48 black stilts over a 3-year period (1997-1999). The cause of death was determined, and thyroid glands were examined histopathologically and compared with those of free-living pied stilts. Concentrations of triiodothyronine (T3) and thyroxine (T4) in the serum of sub-adult and adult stilts were measured before and after iodine supplementation. RESULTS: The main causes of death of captive-reared black stilts following release were trauma, predation and starvation. An increase in size of the thyroid gland due to follicular hyperplasia and dilation was seen in all birds with intact thyroid glands (n=27). Dysplastic follicular changes such as epithelial desquamation, lipid deposition and haemorrhage were common in a large proportion of individuals with goitre. Dietary supplementation with iodine greatly improved survival rates in sub-adults following release, and significantly increased concentrations of T3 and T4 in serum. CONCLUSIONS: Subclinical goitre due to thyroid hyperplasia and dysplasia was the cause of hypothyroidism and this contributed to the poor survival of released sub-adult black stilts raised in captivity. Iodine supplementation of the diet of captive adults and sub-adults resulted in increased concentrations of T3 and T4 in serum and improved survivability.     

Malignant melanoma in a seagull (Larus fuscus): morphological and immunohistochemical approach

A common seagull (Larus fuscus) was found near the southern coast of Italy by the veterinarians of the local wild animal rescue center. Physical examination of the bird revealed an ulcerated mass involving a majority of the oral cavity; the mass did not allow for normal feeding. After the bird died necropsy was performed and the mass was histologically and immunohistochemically examined. The morphology and the immunoreactivity for Melan-A and S-100 antigens led to a diagnosis of malignant melanoma. This is the first case of malignant melanoma described in a seagull, and herein we compare the characteristics of the present case with malignant melanoma already described in domestic animals.