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.     

Evaluation of a Motorized Morcellator for Laparoscopic Removal of Granulosa‐Theca Cell Tumors in Standing Mares

Objective: To describe a motorized morcellator technique for laparoscopic removal of granulosa‐theca cell tumors (GCT) in standing mares and to evaluate long‐term outcome. Study Design: Case series. Animals: Mares (n=7) aged 4–15 years, with unilateral GCT. Methods: Tumor size was determined by transrectal palpation and ultrasonography. Standing sedated mares had 3 laparoscopic portal sites in the paralumbar fossa. After laparoscopic observation of the GCT, the mesovarium was desensitized, the ovarian pedicle transected with a LigaSure^® device, and the ovary grasped with forceps and cut in cylindrical tissue blocks using a motorized morcellator. Tissue blocks were removed through the laparoscopic sleeve. Outcome was determined by telephone interview of owners 6–40 months after surgery. Results: Estimated ultrasonographic median GCT diameter was 17 cm (range, 10–22 cm). Surgical time was 2–4.5 hours. There were no surgical complications. Two mares had mild subcutaneous emphysema at the portals after surgery. Convalescence was short, owners were satisfied with cosmetic outcome, and clinical signs associated with GCT did not recur. Conclusion: The motorized morcellator allows piecemeal removal of large GCT through a relatively small laparoscopic portal. Surgical complications were rare and the cosmetic outcome is favorable. Clinical Relevance: A motorized morcellator is a safe and minimally invasive technique for laparoscopic removal of GCT in mares.     

Histopathological findings in the early diagnosis of granulosa cell tumour in bitches

Granulosa cell tumour (GCT) is a majorly observed ovarian tumour in female dogs. It is essential to diagnose GCT in its initial phase before any symptoms occur, as histological and physiological differences may be observed based on the evolution of this neoplasia. This study aimed to analyse the anatomic histopathology of GCT in its initial stage, with findings of ovaries not yet with the suspicion of neoplasms in the Canis familiaris. A sample including 55 ovaries presented GCT in 40 female dogs. The histopathological analysis was performed considering the intensity of pleomorphism, vascularization and inflammatory infiltrate. Furthermore, we evaluated the mitoses count in 10 fields using 40× magnification. Out of the 40 animals evaluated, 62.5% (25/40) presented the tumour in only one ovary. The Call‐Exner corpuscle was present in 65% (26/40) of the cases. The follicular histological pattern was present in 52.5% (21/40) of the animals. The presence of the Call‐Exner bodies and the degree of tumour cell pleomorphism (p = 0.033) were associated. Moreover, the degree of vascularization and the intensity of the inflammatory infiltrate were also related (p = 0.001). In addition, there was a positive relationship between the increase in pleomorphism and the mean age of the animals (p = 0.044). This study confirmed that the appearance of this tumour may precede any clinical symptomatology. In this study, the most frequent histopathological pattern was the follicular. The characteristics of the granulosa cell tumour diagnosed early were poorly pleomorphic cells, low mitotic index and presence of Call‐Exner body.     

Evaluation of tongue as a complementary sample for the diagnosis of parvoviral infection in dogs and cats.

Diagnosis of canine parvovirus type 2 and feline panleukopenia virus infection in dogs and cats may be hampered by the severity of enteric lesions, secondary bacterial overgrowth, and rapid onset of autolysis. In contrast to small intestine, tongue epithelium is less sensitive to postmortem changes. Sections of tongue and small intestine from 11 dogs and 11 cats with a clinical history and gross and microscopic lesions compatible with canine and feline parvoviral infection were examined for parvoviral infection using real-time polymerase chain reaction (PCR), immunohistochemistry (IHC), and direct fluorescent antibody testing (FA). Parvoviral DNA was detected by PCR in both small intestine and tongue of all but 1 dog. Nineteen of 22 animals (86%) with suspect or positive FA staining in the small intestine also had positive FA and IHC staining in the tongue. Three of 3 dogs (100%) whose carcasses had been frozen and thawed prior to necropsy had more consistently positive staining in tongue than in small intestine by FA and IHC. These data confirm tongue as an excellent complementary sample for parvoviral testing in dogs and cats, especially in cases in which postmortem autolysis has occurred.     

METASTATIC GRANULOSA CELL TUMOR IN A MANGROVE SNAKE (BOIGA DENDROPHILA MELANOTA)

A 12-year-old, female mangrove snake (Boiga dendrophila melanota) was presented with a 1-month history of anorexia, lethargy, and progressive distention of the caudal coelomic cavity. Based on ultrasonographic and cytological examination, an ovarian neoplasia was considered to be the presumptive antemortem diagnosis. Histological examination confirmed the diagnosis of a granulosa cell tumor (GCT) with metastatic tumors in the coelomic fat bodies. Four months following oophorectomy and surgical removal of the distant metastasis, the snake was euthanized because of sudden onset of lethargy, cardiomegaly, and dyspnea. Postmortem examination revealed the presence of metastatic tumors in the lung, liver, and kidneys. Primary ovarian neoplasms are relatively rare in reptiles. Although GCTs have been documented in snakes, definitive diagnosis is typically determined through postmortem histopathological examination. The present case describes a suitable antemortem diagnostic approach of a malignant GCT in a mangrove snake with distant metastases.     

Immunohistochemical and ultrastructural investigation of granular cell tumours in dog, cat, and horse.

Six canine, one feline and one equine granular cell tumours (GCTs) were investigated electron microscopically and immunohistochemically. The tumours were tested for reactivity with monoclonal antibodies against vimentin and desmin and with polyclonal antibodies against cytokeratin, S-100 protein, glial fibrillary acidic protein (GFAP) and neuron specific enolase (NSE). All GCTs were characterized by their PAS positive cytoplasmic granules in light microscopy, which in electron microscopy appeared as lysosome-like granules. In each case two canine GCTs were stained by the antibody against cytokeratin, vimentin and S-100 protein. Cells of the equine GCT showed reactivity with the antiserum against S-100 protein. In the feline GCT no reactivity with any of the antibodies tested was observed. These differences of the immunohistochemical reactions of GCTs suggest a nonuniform histogenesis of GCTs in domestic animals. The reactivity of the tumour cells with the antiserum against NSE is discussed.     

Changes of serum anti‐Müllerian hormone in a mare with granulosa cell tumour following surgery and reinitiation of follicular activity

Anti‐Müllerian hormone (AMH) has been reported to be elevated in mares with granulosa cell tumour (GCT). An 8‐year‐old Thoroughbred mare was presented for not being observed in oestrus after the beginning of the breeding season. Rectal palpation and ultrasonography revealed enlargement and cystic appearance of the left ovary while the right ovary was small with an anoestrous‐like appearance. The AMH concentration was 694.9 ng/ml. Presumptively diagnosed with GCT, the mare was subjected to tumour removal. Histopathology confirmed GCT. To evaluate changes of AMH concentration following surgery, blood samples were collected immediately prior to surgery, and on Days 1, 2, 4, 8, 16 and 32 after surgery. Thereafter, blood samples were collected monthly and also at the time the mare was observed in oestrus (148 days after tumour removal). The AMH concentrations decreased over the first 2 months after surgery (from 721.2 ng/ml to 0.1 ng/ml). Subsequently, AMH concentration increased and peaked at the time of oestrus expression (0.7 ng/ml). The mare then became anoestrous, and AMH concentration decreased and reached 0.2 ng/ml, which was not significantly different from the mean concentration of AMH in normal anoestrous mares (n = 5; 0.26 ± 0.07 ng/ml). In conclusion, the present report implies the potential use of AMH for determination of the time of follicular resumption in mares after GCT removal.     

The relationship between total protein in serum, glutaraldehyde coagulation test and disease in feedlot calves

Serum from in all 249 calves were used to correlate the level of total protein (serum-Tp), gamma-globulin and the reaction of glutaraldehyde coagulation test (GCT) to each other. Further, of the 249 calves 219 were bled within two days after arrival to a feedlot and serum-Tp and GCT were related to subsequent diseases. The results revealed a close relationship between the three tests. Of the 219 calves 15% had a negative GCT (hypogammaglobulinemia) and 26% had a serum-Tp below 55 g/l. There was a significantly (p less than 0.001) higher disease frequency, 81%, in the GCT negative calves than in the GCT positive, 35%. Serum-Tp was significantly (p less than 0.001) higher in healthy compared to diseased calves. The mean values were 60.6, 57.2, 55.2 and 53.1 g/l for healthy calves, calves with respiratory disease, enteritis and both respiratory disease and enteritis respectively. Further, calves with a serum-Tp below 55 g/l developed disease earlier after arrival to the feedlot than those above this limit. It was concluded that both serum-Tp and GCT can be used in field investigation to identify calves predisposed to disease.     

Treatment of haemoperitoneum secondary to ruptured granulosa cell tumours in two mares

This report describes two cases of successful surgical management of granulosa cell tumours (GCT) in mares presenting with haemoperitoneum (HP). Controlled abdominal drainage was initially attempted in Case 1 but was not successful. A ventral midline exploratory laparotomy allowed removal of a haemorrhaging 13 kg GCT. The mare made a full recovery and returned to normal work as a driving pony 11 months post-operatively. In Case 2 controlled abdominal drainage was followed by standing left flank laparoscopic visualisation of the bleeding ovary and transection of the ovarian pedicle by electrocautery. The GCT was then removed via a ventral midline incision due to its large size. Haemoperitoneum can be associated with GCTs and in some cases is severe enough to prompt emergency treatment. Stabilisation of the patient and removal of the haemorrhaging GCT can lead to a successful outcome.     

Cytologic findings from a benign giant cell tumor of the tendon sheath in a dog

A 6‐year‐old male neutered Australian Shepherd dog was presented for evaluation of a subcutaneous mass on the plantar aspect of the proximal left metatarsus. Fine‐needle aspirate smears contained numerous plump spindle cells and large multinucleated cells amongst a considerable amount of pink extracellular matrix. Histopathologic diagnosis of the tissue obtained during initial biopsy and eventual surgical cytoreduction of the mass was a benign giant cell tumor of the tendon sheath (GCTTS). Immunohistochemically, the synovioblastic neoplastic cells were diffusely strongly positive for vimentin and S‐100, were multifocally moderately positive for cytokeratin AE1/3, and were negative for CD18, muscle‐specific actin (MSA), and melanoma‐associated antigen (mutated) 1 (MUM‐1). The dog recovered from surgery and underwent definitive radiation therapy to treat the local residual disease. Eight months later, the mass had not recurred. The diagnosis of GCTTS in this case supports previously published reports describing GCTTS as a relevant disease entity in dogs, and provides the first documentation of cytologic findings with this tumor. Further investigation is needed to correlate pathologic features with clinical behavior and response to therapy in dogs.     

1例猫肾上腺皮质癌脾转移的病理及免疫组化分析

猫的肾上腺皮质癌转移至脾在兽医临床非常罕见。在本报告中,描述了1例10岁田园猫,其症状表现为腹部疼痛,频繁呕吐,消瘦,有慢性肾上腺功能损伤病史,影像学检查脾肿大。对患猫进行脾摘除手术,并对摘除的组织进行组织病理学检查和免疫组化分析。病理组织学结果显示,肿瘤细胞具有丰富的脂质样空泡,细胞体积大,有丝分裂象多见,且肿瘤细胞已发生血管侵袭,细胞核浓染;免疫组织化学结果显示,synaptophysin、S-100和GATA4阳性表达。根据病理组织学和免疫组织化学及病史和实验室检查,最后诊断为肾上腺皮质癌的脾转移。首次报道了猫肾上腺皮质癌脾转移临床病例及病理学诊断,对于临床相关疾病诊疗有一定的参考价值和借鉴意义。     

Spontaneous Basal cell carcinoma of the submandibular gland in a rat

At necropsy, a white nodule (about 5 × 3 mm in size) was observed in the right submandibular gland of a 10-week-old female GALAS rat. Histopathologically, oval to spindle-shaped and pale basophilic tumor cells proliferated closely, and formed variably sized foci. The nodule partially spread into or invaded the surrounding normal tissue, and necrotic foci were recognized in the tumor. Immunohistochemically, the nuclei of the tumor cells showed a diffusely positive reaction for p63, and the cytoplasm showed a diffusely positive reaction for cytokeratin and negative reaction for αSMA, vimentin, desmin and S-100. Many tumor cells were positive for PCNA. Ultrastructurally, the tumor cells contained many tonofilaments in the cytoplasm and a few desmosomes at the intercellular portion. Based on these findings, the tumor was diagnosed as a basal cell carcinoma originating from the duct in the rat submandibular gland.     

Immunohistochemical and clinical evaluation of p53 in canine cutaneous mast cell tumors

One hundred twenty-six cutaneous mast cell tumors obtained by excisional biopsy from 106 dogs were evaluated using immunohistochemical staining for the presence of p53 protein. A standard avidin-biotin immunohistochemical protocol was used incorporating a polyclonal antibody of rabbit origin (CM-1) as the primary antibody. Histopathologic grading of tumors was performed on hemotoxylin and eosin-stained samples. There was a significant difference in the percentage of cells staining positive for p53 for the histopathologic grades (P = 0.0005). Grade III tumors had a significantly greater p53 content than did grade I or II tumors (P < 0.05). Clinical data obtained retrospectively was available for 54 dogs. Tumor recurred in 19 of 54 (35.2%) dogs. Twenty-nine dogs died by the end of the study; 9 of 29 (31.0%) died of mast cell tumor disease. Histopathologic grade showed a significant negative association with survival time. Both clinical stage and histopathologic grade showed a significant negative association with time to recurrence. The percentage of cells staining positive for p53 did not significantly improve the forward analysis. Immunohistochemical detection of p53 did not appear useful in characterizing the clinical association between cutaneous mast cell tumor cellular features and survival time or time to tumor recurrence in dogs.     

The utility of intraoperative impression smear cytology of intracranial granular cell tumors: Three cases

Two adult male dogs (a 7‐year‐old shorthaired Chihuahua and 14‐year‐old Shih Tzu) and one adult female dog (a 9‐year‐old Maltese) presented for evaluation of new‐onset seizure activity. Magnetic resonance imaging of the brain demonstrated a large, poorly marginated T2‐weighted hyperintense, and strong contrast enhancing extra‐axial mass in each case. A surgical biopsy for histopathologic evaluation was elected in all cases, and intraoperative impression smears were successfully obtained. Intraoperative cytology identified a homogenous population of round to polygonal cells with central to eccentric nuclei, coarse chromatin, and variably amphophilic to eosinophilic granular cytoplasm. Cytologic findings led to a suspected diagnosis of granular cell tumor (GCT) in all cases. Histopathologic review identified a densely cellular, unencapsulated neoplastic mass comprised of sheets of large round to polygonal cells with abundant eosinophilic cytoplasm containing numerous eosinophilic intracytoplasmic granules, confirming the diagnosis of GCT in all cases. The cases reported here are unique in that they reveal an accurate intraoperative cytologic diagnosis of a rare canine central nervous system neoplasm. Intraoperative cytology of the intracranial masses could provide clinicians with important and quick diagnostic and prognostic information; therefore, expediting decisions made intraoperatively. Further research is warranted to determine the diagnostic accuracy of intraoperative cytology for neoplasia in veterinary patients.     

Tongue rotation for reconstruction after rostral hemiglossectomy for excision of a liposarcoma of the rostral quadrant of the tongue in a dog

A 15-year-old female beagle dog was presented for a lingual liposarcoma. Full staging of the disease did not show any evidence of metastasis. A tongue rotation for reconstruction after rostral hemiglossectomy was performed after removal of the mass. This surgical technique may be useful in tongue reconstruction after trauma or tumor excision.     

Determination of serum anti‐Müllerian hormone concentrations for the diagnosis of granulosa‐cell tumours in mares

Reasons for performing study: Endocrinological assays are important for evaluation of mares with granulosa‐cell tumours (GCTs), and our research in mares indicates that anti‐Müllerian hormone (AMH) may be a good biomarker for this type of ovarian tumour. Objectives: To evaluate the use of serum AMH concentrations for endocrine diagnosis of GCTs in mares. Methods: Archived serum samples (n = 403) previously assayed for determination of serum inhibin, testosterone and progesterone concentrations (GCT panel) were assayed for serum AMH concentrations using a heterologous enzyme‐linked immunosorbent assay previously validated by our laboratory. For a subset (n = 44) of these samples, a clinical diagnosis of GCT was confirmed by histopathology. Results: Overall, the sensitivity of AMH (98%) for detection of histologically confirmed GCTs was significantly (P<0.05) greater than that of either inhibin (80%) or testosterone (48%) or the combination of inhibin and testosterone (84%). Conclusions: Determination of serum AMH concentrations is a useful biomarker for detection of GCTs in the mare. Potential relevance: Measurement of serum AMH concentrations can be used for diagnosis of GCTs in the mare. As serum AMH concentrations do not vary significantly during the oestrous cycle or pregnancy, interpretation of these results is not confounded by these physiological states.     

A peripheral primitive neuroectodermal tumor with generalized bone metastases in a puppy

A peripheral primitive neuroectodermal tumor (pPNET), most consistent with a human Ewing's sarcoma, is described in a 5-month-old male Australian Shepherd puppy. The first tumor site detected was in the left frontal bone of the skull with apparent subsequent rapid metastases to multiple sites in the axial and appendicular skeleton and bone marrow, kidneys, and perihyphophyseal meninges. Radiographically, all bone lesions were lytic and there was also a humeral bone fracture. Histologically, the tumor was diagnosed as a small round blue cell tumor. At this stage, the differential diagnosis included a lymphoma, rhabdomyosarcoma, and a PNET of the peripheral nervous system. However, the cells had positive expression of triple neurofilament antigens as detected immunocytochemically. The cells were negative for a broad panel of canine-specific leucocyte cell marker antigens for desmin, smooth muscle actin, synaptophysin, and CD99. Ultrastructurally, the cells contained occasional dense core neurosecretory granules and intermediate filaments with intercellular desmosomal-like junctions and abundant glycogen clusters. Based on the age of the dog, the clinical history, the distribution of gross lesions, histologic characteristics of a small round blue cell tumor, and immunocytochemical and ultrastructural evidence of neuroectodermal differentiation, a diagnosis of a pPNET similar to a human Ewing's sarcoma was made.     

Symptomatic granular cell tumor involving the pituitary gland in a dog: a case report and review of the literature

A granular cell tumor involving the pituitary gland, optic chiasm and ventral pyriform lobes was discovered in a 12-year-old Labrador Retriever. Clinical signs included acute blindness, seizures, ataxia, weakness, and behavioral changes. The diagnosis was established by histopathologic and ultrastructural examination of neoplastic tissues collected at necropsy. Granular cell tumors involving the central nervous system are well documented in humans but rarely have been described in dogs. The location of the neoplasm and the clinical symptoms seen in this dog closely parallel those of a rare syndrome in humans commonly described as symptomatic parasellar or pituitary granular cell tumors. The cell of origin for these tumors is still highly debated, and attempts to characterize human granular cell tumors through immunohistochemistry have produced conflicting results. An immunohistochemical profile of this neoplasm revealed focal positive staining for vimentin with a lack of staining for neuron-specific enolase, glial fibrillary acidic protein, S-100, and synaptophysin. All neoplastic cells were strongly positive with the periodic acid-Schiff reaction.     

Feline periocular peripheral nerve sheath tumor: a case series

The objective of this retrospective study was to describe signalment, clinical presentation, surgical treatment, outcome, histopathologic, and immunohistochemical characteristics of periocular peripheral nerve sheath tumor in a series of cats. Six cats with a histologic diagnosis of peripheral nerve sheath tumor of the eyelid and/or conjunctiva were identified from 3997 feline ocular histopathologic submissions to the Comparative Ocular Pathology Laboratory of Wisconsin between 1976 and 2002. Medical records were reviewed and data were collected for signalment, eye affected, location, duration, and history of tumor recurrence. Ancillary diagnostic tests, surgical approach, and histopathology were also described. Special stains and immunohistochemistry panels were performed on the tissue. Follow-up information was obtained through telecommunications with the owner and local veterinarian. The mean age of the cats was 8 years (range: 5-15 years). There was an equal distribution of males and females. Breeds represented were two Domestic Long-haired (DLH) cats, three Domestic Short-haired (DSH) cats, and one Manx. The right eye was affected in four cats and the left eye in two cats. The superior eyelid was affected in four cats, the superior conjunctiva in two cats, and the inferior eyelid in one cat. All of the cats that underwent surgical excision, cryoablation, or laser ablation had an average of three recurrences of the tumor. Four of six cats undergoing wide excision with enucleation, exenteration or rhomboid flap had no recurrence of the tumor. Histomorphologically, all of the tumors were of spindle cell type with five of six tumors showing histologic features of Antoni type B tissue. All six tumors stained positive for vimentin. Five of six tumors stained positive for S-100 and laminin. All six tumors were negative for smooth muscle actin. Definitive histologic diagnoses of spindle cell tumors can be difficult. The presence of Antoni A and B histologic patterns as well as immunohistochemical reactivity for vimentin, S-100, and laminin suggest a diagnosis of peripheral nerve sheath tumor. Our findings suggest that local aggressive recurrence is common and that wide surgical excision combined with enucleation or exenteration may be indicated.     

A case of papillary squamous cell carcinoma in the mandible of a young French bulldog

A 7-month-old castrated French bulldog was presented with a left-sided mandibular tumor. The initial tumor biopsy diagnosis was ameloblastoma. The owner brought this dog the Kitasato University Veterinary Teaching Hospital for more detailed examination and treatment. Computed tomography revealed a tumor on the left lateral mandibular gingiva from the caudal third of the incisor tooth to the right canine tooth, associated with severe amorphous osteolysis of the mandibular bone. The tumor was surgically excised and diagnosed as papillary squamous cell carcinoma. Currently, 2514 d (6.9 y) since the operation, the dog is healthy, without recurrence.Key clinical message:Although papillary squamous cell carcinoma is rare, many cases have been reported in the oral cavity of medium-to large-sized dogs. Based on this report, papillary squamous cell carcinoma can occur in small dogs such as young French bulldogs and a good prognosis can be achieved with proper resection.