Bilateral lymphosarcoma of the nictitating membrane in a Quarter Horse

A 21-year-old Quarter Horse mare presented for a mass of the right nictitating membrane. The entire right nictitating membrane was surgically removed and diagnosed as a mixed cell lymphosarcoma and squamous cell carcinoma. The horse had no systemic signs of lymphosarcoma. Approximately 1.5 years later, the horse presented with a similar mass in the left nictitating membrane. The entire left nictitating membrane was surgically removed and diagnosed as a mixed cell lymphosarcoma. In this case, complete surgical removal of the masses resulted in a cure. The horse has remained free of systemic lymphosarcoma for over 3 years.

Case report

A 21-year-old gray Quarter Horse mare was presented to the Kansas State University Veterinary Medical Teaching Hospital for evaluation of a mass involving the right nictitating membrane. The mass was first noticed 5 weeks before presentation and had been treated by the referring veterinarian with a topical steroid solution twice daily. Administration of the topical steroid caused some decrease in the swelling, but it quickly returned once medication was discontinued. The horse had no other medical complaints and appeared to be otherwise healthy. On ocular examination, chemosis of the third eyelid and conjunctiva of the right eye was evident. Direct and indirect pupillary light reflexes, Schirmer tear test, and intraocular pressures were normal in both eyes. Fluorescein stain uptake was negative in both corneas. Cranial nerve and vision examinations did not reveal any abnormalities. On further examination of the right nictitating membrane, a firm mass was palpated under and protruding from the palpebral conjunctiva (Fig 1). No other abnormalities were found on physical examination.     

Corneal invasion by hemangiosarcoma in a horse

A 15-year-old gray Arabian gelding presented for evaluation of a lateral limbal mass extending across approximately 30% of the cornea. Grossly, the raised mass appeared nonpigmented, smooth, and irregular in shape, with an area of central necrosis and serosanguinous discharge. The mass was removed via lamellar keratectomy and histopathologic evaluation revealed features characteristic of hemangiosarcoma (HSA), including irregular vascular channels lined by a plump spindle cell population. Immunohistochemistry results showed that the neoplastic cells lining the vascular channels present diffuse and strong cytoplasmic reaction with von Willebrand Factor and the perivascular spindle cells exhibit moderate cytoplasmic reaction for smooth muscle actin. A lack of cytokeratin staining definitively excluded a diagnosis of atypical squamous cell carcinoma. Smooth muscle actin staining of the perivascular cells adjacent to the neoplastic endothelial cells is not a feature commonly described in HSA and has not been reported in previous cases of equine HSA. The horse remained in good health 21 months postkeratectomy and has exceeded the survival time of previously documented equine ocular HSA cases where more extreme surgical excision was performed.     

Keratoconjunctivitis sicca: histopathologic study of nictitating membrane and lacrimal glands from 28 dogs

Forty nictitating membrane glands and 9 main lacrimal glands were obtained for histologic evaluation from 28 dogs with keratoconjunctivitis sicca as the result of azosulfapyridine toxicity, canine distemper, multisystemic autoimmune disease (Sj?gren's syndrome-like syndrome), congenital origin, and unilateral and bilateral idiopathic keratoconjunctivitis sicca. Similar glands from 6 control dogs were studied. The most prevalent (87%) histopathologic finding was variable degrees of multifocal chronic adenitis, characterized by acinar atrophy and replacement with increased numbers of plasma cells and lymphocytes within increased amounts of interacinar fibrous connective tissue stroma. Occasional tubular structures were dilated and filled with neutrophils and cellular debris.     

Lymphangiosarcoma in a dog treated with surgery and chemotherapy

A large subcutaneous mass at the left cervical site in a 9-year-old male Siberian husky was removed surgically. Histopathologically, the mass was mainly consisted of a proliferation of spindle-shaped neoplastic cells arranging in solid sheath and partially vascular channels containing few blood cells. The tumor cells exhibited highly invasive activity to the surrounding tissues. In addition, the tumor cells were immunopositive for Factor VIII-related antigen. On the basis of these findings, the tumor was diagnosed as lymphangiosarcoma. Recurrent mass was noticed 3 weeks after surgery but completely disappeared after the doxorubicin treatment. Neither recurrence nor metastasis was observed for 9 months after the remission.     

Immunohistochemical detection of scrapie prion proteins in clinically normal sheep in Pennsylvania.

Following diagnosis of scrapie in a clinically suspect Suffolk sheep, 7 clinically normal flockmates were purchased by the Pennsylvania Department of Agriculture to determine their scrapie status using an immunohistochemical procedure. Two of the 7 euthanized healthy sheep had positive immunohistochemical staining of the prion protein of scrapie (PrP-Sc) in their brains, nictitating membranes, and tonsils. The PrP-Sc was localized in the areas of the brain where, histopathologically, there was neurodegeneration and astrocytosis. The PrP-Sc occurred within germinal centers of the affected nictitating membranes and tonsils and was located in the cytoplasm of the dendrite-like cells, lymphoid cells, and macrophages. These results confirm that immunohistochemical examination of the nictitating membrane can be used as a screen for the presence of scrapie infection in clinically normal sheep at a capable veterinary diagnostic laboratory. In sheep with a PrP-Sc-positive nictitating membrane, the diagnosis of scrapie should be confirmed by histopathology and immunohistochemical examination of the brain following necropsy. Following full validation, immunohistochemistry assays for detection of PrP-Sc in nictitating membrane lymphoid tissues can improve the effectiveness of the scrapie control and eradication program by allowing diagnosis of the disease in sheep before the appearance of clinical signs.     

Accessory nictitating membrane in a Lipizzaner colt

A 2‐year‐old Lipizzaner colt presented for removal of a mass over the left eye. The colt had no blepharospasm or significant ocular discharge, but the cornea underlying the mass was mildly oedematous. The mass protruded from the conjunctiva at the dorsotemporal aspect of the globe and was covered in normal conjunctiva. It had a gross appearance similar to the cartilaginous flap of a nictitating membrane, but in an aberrant location. It was moveable to cover the dorsal aspect of the cornea when the globe was retropulsed. Excision of the mass under general anaesthesia was elected. Histopathology supported a diagnosis of an accessory nictitating membrane, consisting of a cartilaginous band surrounded by glandular epithelium. The colt recovered from surgery without complication and no problems were reported by the owner at 8.5 months post‐operatively. To the author's knowledge, there have not been previous reports of accessory or ectopic nictitating membranes in equine or other species.     

Successful use of topical mitomycin C for the treatment of a putative neoplastic vascular mass on the nictitating membrane of a dog

A nine-year-old female desexed Great Dane presented with a painful, proliferative, soft red putative neoplastic vascular mass on the nictitating membrane. Three 7-day cycles of the topical cytotoxic drug mitomycin C 0.04%, applied four times daily to the lesion using a low-dose alternate-week pulse therapy, brought about rapid remission of the lesion. The lesion was still in remission at time of euthanasia some 13 months later.     

Synovial hemangioma in the stifle joint of a dog

Synovial hemangioma was diagnosed in an 8-year-old castrated male Belgian Sheepdog with lameness of 3 months' duration. Pain, soft-tissue swelling, and hemarthrosis were localized to the left stifle joint. Projections of synovial membrane with reddish-purple nodules, excised via arthrotomy, were composed histologically of variably sized vascular channels that were lined by well-differentiated endothelial cells and separated by fibrous septa. Distension of some channels resulted in endothelial disruption, thrombosis, hemorrhage, necrosis, and focal spindle-cell proliferation. The limb was amputated to remove remaining neoplastic tissue. The hemangioma extended focally into the joint capsule and popliteal soft tissue but did not invade skeletal muscle or bone. The dog was free of detectable neoplasia 6 months after amputation. Synovial hemangioma is a rare benign vascular proliferation in people, most commonly in the knee, and should be included in the differential diagnosis for canine synovial tumors.     

Treatment of corneal ulceration and bullous keratopathy using a nictitating membrane flap in two horses

Objective

The aim of this study was to describe placement of a nictitating membrane flap as a treatment for corneal ulceration and bullous keratopathy in two horses.

Animals Studied

A 13-year-old American Saddlebred mare presented for severe corneal edema, superficial stromal ulceration, and a central bulla of the left eye. A 4-year-old Trakhener stallion also presented with a large axial bulla of the left eye with concurrent severe corneal edema and a deep stromal ulcer.

Procedure

A complete ophthalmic examination was performed. Samples were obtained for corneal cytology, and both horses were started on aggressive medical therapy. Both underwent general anesthesia for placement of a nictitating membrane flap and a subpalpebral lavage system (SPLS).

Results

Corneal cytology for each horse revealed a mixed bacterial population. Moderate Pseudomonas aeruginosa was cultured from the mare, while Aspergillus species and a few Enterococcus gallinarum were cultured from the stallion. The bullae in both horses resolved at 3 and 4 weeks and vision returned in the affected eye 4.5 and 3 months postoperatively at the last follow-up, respectively.

Conclusion

Aggressive medical management with concurrent placement of a nictitating membrane flap is effective to treat bullous keratopathy in two horses. The described treatments could be used to treat horses that develop severe or progressive bullous corneal lesions.     

Spontaneous Pulmonary Hemangiosarcoma in a Norway Rat (Rattus norvegicus)

A second parity, 1-year-old female Norway rat (Rattus norvegicus) presented for veterinary evaluation 5 weeks postpartum with a progressively decreased appetite, weight loss, lethargy, and labored breathing. On physical examination, the animal was in fair body condition, dyspneic, and dehydrated. Owing to a poor prognosis, the owner opted for palliative care for the animal, declined further diagnostic testing, and requested for the rat to be discharged from the hospital. The following day the animal was found dead in its cage, and the body was submitted for necropsy. Gross observations included numerous, variably sized, soft, dark red pulmonary nodules concurrent with hemothorax and hemopericardium. Microscopically, the pulmonary architecture was effaced by nodules of variably sized, irregular, vascular channels lined with neoplastic endothelial cells. The cytoplasm of neoplastic cells in the lung was immunohistochemically positive for von Willebrand factor (i.e., factor VIII–related antigen) and was more intensively and more often positive for vascular endothelial growth factor as compared with nonneoplastic cells. Ultrastructurally, vascular channels were formed by spindloid neoplastic cells with elongated, fused, and interdigitated processes. Based on these findings, a diagnosis of spontaneous pulmonary hemangiosarcoma was made in this Norway rat.     

Local immune response in the chicken Harderian gland to antigen given by different ocular routes.

The effectiveness of three ocular routes of antigen administration to produce a local immune response in the Harderian gland was studied. The routes were by eyedrop, injection into the ocular conjunctiva and injection into the nictitating membrane. The antigen was observed in the cytoplasm of macrophages located within the lymphoid tissue only after the injection into the nictitating membrane. The numbers of germinal centres and plaque forming cells found in the gland after injection into the nictitating membrane was higher than the numbers observed following the other two ocular applications. These findings indicate that the injection of the antigen into the nictitating membrane is the most effective ocular route for producing a local immune response in the Harderian gland.     

Evaluation of a soluble sustained-release ophthalmic delivery unit in the dog.

A 5-mg hydroxypropyl cellulose soluble ocular insert was evaluated by placement in the upper and the lower conjunctival and bulbar nictitating membrane fornices in normal dogs. The insert was easily placed in the conjunctival fornices, but was difficult to place in the bulbar nictitating membrane fornix. The conjunctival inserts did not produce irritation of the cornea and the conjunctiva. Placement of the inserts beneath the nictitating membrane produced local irritation and chemosis. The sites of highest to lowest retention for the insert were upper conjunctival fornix, bulbar nictitating membrane fornix, and lastly, lower conjunctival fornix. Dissolution was nearly complete in 8 hours.     

Lymphangiosarcoma with bone formation of the auricle in a dog

A 12-year-old mixed-breed neutered female dog was referred with cutaneous tumors at the left auricle. Histologically, the cutaneous tumor located in the dermis comprised numerous clefts and cavernous channels lined by neoplastic endothelial cells with no erythrocytes. Bone tissue without direct contact with neoplastic cells was seen in the well-developed stromal connective tissue. The neoplastic endothelial cells exhibited mild to moderate atypia. Immunohistochemically, neoplastic cells were positive for vimentin and negative for cytokeratin and factor VIII-related antigen. Basement membrane around the neoplastic lumens was positive for laminin in a linear or granular pattern. Ultrastructural examination revealed discontinuous basement membrane beneath the tumor cells. Histopathological features of this case were consistent with lymphangiosarcoma, and stromal ossification was characteristic.     

Presumed primary ocular lymphangiosarcoma with metastasis in a miniature horse

A 7‐year‐old, 153.0‐kg American Miniature mare presented for evaluation of keratoconjunctivitis of the right eye (OD). A superior palpebral conjunctival mass and stromal keratitis were diagnosed. The incisional biopsy diagnosis was a presumptive corneal hemangiosarcoma. Transpalpebral enucleation was performed, and histopathologic evaluation confirmed angiosarcoma of the conjunctiva, cornea, and extraocular muscles. The horse developed progressive epistaxis and orbital swelling following surgery. A systemic workup was performed 3 months after enucleation, revealing regrowth within the orbit and marked cranial cervical lymphomegaly, suggestive of metastasis. Humane euthanasia was performed, and necropsy confirmed a locally invasive periorbital tumor with metastasis to the submandibular tissue, submandibular lymph node, and thoracic inlet. Histopathologic evaluation of necropsy specimens revealed polygonal to spindle neoplastic cells lining neoplastic vascular channels lacking erythrocytes. Immunohistochemically, the neoplastic cells labeled strongly positive for PROX‐1, vimentin, CD‐31, VEGF, weakly positive for factor VIII‐related antigen, and negative for collagen IV. Based on the clinical, histological, and immunohistochemical features of this tumor, a primary ocular lymphangiosarcoma with metastasis was diagnosed.     

A preliminary study of changes in tear film proteins in the feline eye following nictitating membrane removal

Objective To investigate the influence of nictitating membrane (third eyelid) removal on selected proteins in feline tears. Animal studied Domestic short‐haired cats (7–17 months; 2.6–5.2 kg) were used. Procedures Eye‐flush tears were collected periodically for up to 18 weeks from both eyes of animals with nictitating membranes removed, but nictitating gland left intact, (n = 4) or with nictitating membranes intact (n = 4). Tear comparisons were based on total protein content (TPC) using micro bicinchoninic acid assay, immunoglobulin A (IgA), and matrix‐metalloproteinase (MMP)‐9 measurements using sandwich enzyme‐linked immunosorbent assay (ELISA) and tear gelatinase activity using gelatin zymography. Expression of MMP‐2 and ‐9 in nictitating membranes removed at baseline (week 0) and eyes collected at 18 weeks were also investigated in histological sections using immunoperoxidase for visualization. Results Nictitating membrane removal did not significantly change TPC and MMP‐9 in tears within the first 4 weeks. MMP‐9 was not detected by ELISA in tears from eyes without nictitating membranes from week 5 onwards. IgA (%IgA of TPC) data varied between animals. Gelatin zymography showed increased MMP‐2 and ‐9 activity in tears from eyes without nictitating membranes at week 1 and a decrease following week 2 post‐surgery. MMP‐2 and ‐9 were immunolocalised to conjunctival goblet cells of removed nictitating membranes and to the conjunctival epithelium, respectively. After 18 weeks, the distribution of MMPs in tissue was comparable between eyes with and without nictitating membranes. Conclusions Based on this preliminary study, nictitating membrane removal appeared to cause long‐term changes in expression of tear proteins, including reduced MMP‐9 expression.     

Ocular angiosarcoma in the horse: morphological and immunohistochemical studies

Angiosarcomas arising in ocular tissues of four aging horses are described. Tumors were locally invasive and eventually metastasized via the mandibular and cervical lymph nodes. Pathologically, the tumors contained well-differentiated regions, in which vascular channels were lined by pleomorphic endothelial cells, as well as poorly-differentiated regions, in which vascular channels were either rudimentary or absent. Red blood cells were scare in vascular structures formed by the tumors. Factor VIII related antigen (VIII:RAg), a blood vascular endothelial marker, was demonstrated by immunohistochemistry in tumor cells in both the well-differentiated and poorly-differentiated regions. The results suggest that ocular tumors arose in blood vascular endothelium and represented true hemangiosarcomas although a lymphatic endothelial origin could not be excluded. Immunohistochemical demonstration of VIII:RAg is a valuable adjunct to conventional stains in the diagnosis of equine vascular neoplasia.     

CASE REPORT: Medical and surgical management of severe inflammation of the nictitating membrane in a Giant Panda (Ailuropoda melanoleuca)

A 10‐year‐old male giant panda (Ailuropoda melanoleuca) presented for severe, acute swelling and protrusion of the right nictitating membrane, unresponsive to topical therapy. Excisional biopsy of the nictitating membrane and its associated lacrimal gland was elected due to necrosis and friability of the tissue. Histopathology revealed suppurative, necrotizing conjunctivitis and dacryoadenitis. Culture grew Stenotrophomonas maltophila and Enterococcus spp with extensive antibiotic resistance. Treatment with topical and systemic antibiotics based on sensitivity results was initiated. All treatments were well tolerated. Healing was uncomplicated with no recurrence of the lesion and no clinical evidence of keratoconjunctivitis sicca. Ophthalmic exams and Schirmer tear tests performed opportunistically during postoperative anesthetic procedures were unremarkable, confirming that excision of the nictitating membrane had not caused clinically significant detrimental effects. The etiology of this lesion remains undetermined, but trauma is suspected. To the authors’ knowledge, this is the first report of nictitating membrane pathology and excision in the giant panda.     

Surgical management of orbital nodular granulomatous episcleritis in a dog

A 27 kg, 6‐year‐old, male castrated German shorthaired pointer presented to the University of Missouri, Veterinary Teaching Hospital with the complaint of progressive exophthalmia of 2 years duration optical density (OD). Lack of retropulsion OD was noted on physical examination. Anterior segment examination OU and fundic examination OS did not reveal any abnormalities. Examination of the fundus OD revealed focal scleral indentation of the inferior nasal globe. The indentation changed location with globe movement OD. MRI and CT scan revealed a well‐circumscribed, approximately 2 cm in diameter mass located caudal and ventral to the affected globe that appeared to communicate with the nictitating membrane with absence of any bony involvement. A modified lateral orbitotomy was recommended and performed to remove the orbital mass and nictitating membrane en‐bloc. Histopathology and immunohistochemistry of the mass confirmed a diagnosis of nodular granulomatous episcleritis (NGE). Postoperatively, the dog developed absolute keratoconjunctivitis sicca (KCS). Examples of primary episcleral inflammation in the dog include diffuse episcleritis, NGE, nodular fasciitis, fibrous histiocytoma, proliferative conjunctivitis/keratoconjunctivitis, pseudotumor, and Collie granuloma. The etiology of these episcleral inflammations is presumed to be immune mediated. To our knowledge, this is the first report of NGE affecting the orbital region of a dog. Development of absolute KCS resulting from excision of the nictitating membrane is also supported by this case.     

Feline conjunctival hemangioma and hemangiosarcoma: a retrospective evaluation of eight cases (1993-2004)

The purpose of this retrospective study was to evaluate feline primary conjunctival vascular tumors of endothelial origin. Eight cases (six hemangiomas, two hemangiosarcomas) from a collection of 3460 feline submissions between 1993 and 2004 were evaluated using routine hematoxylin and eosin (H&E). Signalment, location, size, duration, epithelial pigmentation, margins, adjuvant therapy, outdoor activity, and geographic location, comparing ultraviolet (UV) radiation levels by state, were recorded. Follow-up information was available for five cases. In this study, the average age was 10.6 years, with neutered males over-represented. The Domestic Short-haired cat was most commonly affected. The most common anatomic site was the nictitating membrane, with the left eye preferentially affected. The average size and duration, prior to presentation, was 7.5 mm and 4.4 months, respectively. Seven of eight cases were devoid of epithelial pigmentation in nonaffected areas and the majority of cases were from states with high annual UV-light exposure. Only cases of hemangiosarcoma underwent surgical re-excisions following incomplete excision; however, no further recurrences were reported. No cases evaluated had evidence of metastatic disease at the time of excision. Surgical excision alone may be curative. However, recurrence is possible. These tumors demonstrate similar predilection sites and involvement of nonpigmented epithelium, as is true in canine cases, which may relate to risk factors as well.     

Symblepharon with aberrant protrusion of the nictitating membrane in the snowy owl (Nyctea scandiaca)

Two young snowy owl chicks were presented with aberrant protrusion of the nictitating membranes. This was caused by conjunctival adhesions causing symblepharon secondary to a previous septicemia episode. While symblepharon has been noted in birds before, this unusual presentation of the nictitating membrane has not been reported. Surgical intervention ameliorated the clinical signs, allowing vision in one bird by removal of the nictitating membranes, a technique which appeared to have no deleterious effects on the ocular surface.