Evaluation of matrix metalloproteinases and tissue inhibitors of metalloproteinases in aqueous humor of dogs with versus without naturally occurring primary angle-closure glaucoma.

OBJECTIVE: To compare concentrations of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in aqueous humor from ophthalmologically normal dogs and dogs with naturally occurring primary angle-closure glaucoma (cPACG). SAMPLE: Aqueous humor samples from 12 eyes with cPACG and 18 ophthalmologically normal eyes of dogs. PROCEDURES: A multiplex fluorescence-based ELISA was used to measure concentrations of MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, MMP-10, MMP-13, TIMP-1, TIMP-2, and TIMP-4. Results for eyes with versus without cPACG were compared. RESULTS: Significantly higher mean concentrations of MMP-1 (45% higher), MMP-2 (55% higher), MMP-3 (39% higher), MMP-8 (79% higher), MMP-9 (29% higher), MMP-10 (60% higher), TIMP-1 (63% higher), and TIMP-2 (136% higher) were detected in aqueous humor from eyes with cPACG, compared with ophthalmologically normal eyes. CLINICAL RELEVANCE: MMPs and TIMPs have pivotal roles in extracellular matrix turnover and homeostasis in the outflow pathways of the eye. Results of the present study documented higher concentrations of MMPs and TIMPs in aqueous humor samples from dog eyes with late-stage cPACG. Although, to our knowledge, TIMPs have not previously been evaluated in the context of cPACG, the markedly higher concentration of TIMPs in eyes with cPACG suggested that inhibition of proteolysis and extracellular matrix turnover might be a factor in the development of glaucoma in susceptible individuals. However, because the present study used samples from dogs with late-stage cPACG, further work is required to characterize the temporal relationship between MMP and TIMP concentration changes and onset or progression of disease.

Assessment of a New Antigen Detection Test for the Diagnosis of Canine Visceral Leishmaniasis.

Canine visceral leishmaniasis (CVL) is a serious zoonotic disease in Brazil and Southern Europe. CVL is primarily caused by Leishmania infantum and its diagnosis relies largely on detection of parasites in bone marrow or lymph node aspirates by microscopic observation of the parasites in stained smears, parasite culture, or polymerase chain reaction (PCR). Serological tests exist but they do not distinguish active disease from simple exposure to parasite antigens. Here, we have assessed the utility of a new monoclonal antibody--based antigen (protein) detection test for the diagnosis of CVL. The test was positive in 70% of beagle dogs experimentally infected with L. infantum. In contrast, culture of the parasites from bone marrow aspirates was positive in only 40% of the infected animals. These preliminary results suggest that this antigen detection test, which we have recently described for the diagnosis of human VL, has the potential to be a useful diagnostic tool for CVL.

Evaluation of cytology and histopathology for the diagnosis of feline orbital neoplasia: 81 cases (2004-2019) and review of the literature.

OBJECTIVE: To provide an updated overview of feline orbital neoplasia, to compare diagnostic utility of cytology and histopathology, and to evaluate minimally invasive sampling modalities. PROCEDURES: A medical records search was performed to identify cats with orbital neoplasia. Data were collected regarding signalment, diagnosis, vision status, imaging modalities, and sample collection methods. A reference population with orbital neoplasia was also identified via literature search for comparison with regard to final diagnosis. RESULTS: Eighty-one cats met selection criteria and 140 cases were identified in the literature. In the study and reference populations, respectively, diagnoses were grouped as follows: round cell tumors 47% and 24%, epithelial tumors 38% and 40%, mesenchymal tumors 14% and 34%, and neurologic origin tumors 1% and 2%. The most common diagnoses in both groups were lymphoma and squamous cell carcinoma (SCC). Feline restrictive orbital myofibroblastic sarcoma (FROMS) was common in the reference population but not diagnosed in the study population. Cytology results were available for 41 cats; histopathology results were available for 65 cats. Both cytology and histopathology results were available for 25 cats, in 44% of which cytologic results were overturned. No significant complications were associated with any sampling method. Lack of cats with multiple samples available for histopathology limited comparison between tissue sampling methods. CONCLUSIONS: Orbital neoplasia is common in cats, with round cell and epithelial tumors diagnosed most commonly in the study population. Histopathology is superior to cytology in providing a definitive diagnosis. Minimally invasive tissue biopsy techniques appear to be safe and effective.

Evaluation of cytology and histopathology for the diagnosis of canine orbital neoplasia: 112 cases (2004-2019) and review of the literature.

OBJECTIVE: To provide an updated overview of canine orbital neoplasia, to compare diagnostic utility of cytology and histopathology, and to evaluate alternative sampling modalities, particularly image-guided core needle biopsy. PROCEDURES: A medical records search was performed to identify dogs with orbital neoplasia. Data were collected regarding signalment, diagnosis, vision status, imaging modalities, and sample collection methods. A reference population with orbital neoplasia was also identified via literature search for comparison with regard to final diagnosis. RESULTS: One hundred and twelve dogs met selection criteria. In the study and reference populations, respectively, diagnoses were grouped as follows: mesenchymal tumors 40% and 35%, epithelial tumors 35% and 18%, tumors of neural origin 8% and 37%, and round cell 17% and 10%. The most common diagnoses in the study group were nasal adenocarcinoma, osteosarcoma, lymphoma, and meningioma. Cytology results were available for 47 dogs and histopathology results were available for 95 dogs. Both cytology and histopathology results were available for 30 dogs, in 53% of which results were discordant. Cytology samples were nondiagnostic or provided a diagnosis that was later overturned in 32% of cases in which they were obtained. Results from core needle biopsy samples were nondiagnostic or overturned by surgical biopsy results in only 13% of cases. No significant complications were associated with any sampling method. CONCLUSIONS: Orbital neoplasia is common in dogs. Histopathology is superior to cytology in providing a definitive diagnosis. Image-guided core needle biopsy appears to be a safe and effective means of obtaining samples.

Use of cyanoacrylate adhesive in the surgical management of feline corneal sequestrum: 16 cases (2011-2018).

OBJECTIVE: To evaluate the use of cyanoacrylate adhesive as an adjunct to lamellar keratectomy in cats with corneal sequestrum. METHODS: Medical records were reviewed to identify cats with naturally occurring midstromal corneal sequestra treated with lamellar keratectomy and cyanoacrylate adhesive. All cats also had a bandage contact lens placed for postoperative comfort. Data collected included breed, age, sex, and reproductive status of the cat, eye involved, presence or absence of neovascularization at the time of surgery, history of prior sequestra, additional ocular procedures performed, use of neuromuscular blocking agents, procedure time, time to cessation of topical medications, time to recurrence or last follow-up, and complications aside from recurrence. RESULTS: Sixteen cats met study criteria, with a median follow-up time of 17.5 months in those without recurrence. Median time to cessation of topical medications was 4 weeks. Fourteen cats (87%) have not experienced recurrence. Other than recurrence in two cats, no significant complications were noted. Aside from purebred status, no commonalities were found between the two cats with recurrence. Neuromuscular blocking agents were not used during surgery in most cases. Median procedure time was 10 minutes. CONCLUSIONS: Cyanoacrylate adhesive is an effective and safe alternative to grafting procedures in cats undergoing lamellar keratectomy for treatment of corneal sequestrum. Recurrence rates are comparable to those seen with grafting techniques. Benefits of this approach include decreased anesthesia time, lower procedure costs, and short duration of postoperative treatment.

Use of a nictitating membrane flap for treatment of feline acute corneal hydrops-21 eyes.

OBJECTIVE: To evaluate the effectiveness of the use of a nictitating membrane flap (NMF) as therapy in 19 cats (21 eyes) affected with feline acute corneal hydrops (FACH). METHODS: Medical records from 19 cats diagnosed with FACH and treated with a NMF were retrospectively evaluated. Information was collected from multiple veterinary hospitals and included signalment, medical history, therapy, and ocular outcome. RESULTS: Breeds included 13 Domestic Shorthairs, 2 Exotic Shorthairs, 2 Maine Coons, 1 Persian, and 1 Domestic Medium Hair. Two cats were bilaterally affected. Median age of cats was 3.2 years (range 0.26-15 years). Eleven patients were spayed females, 6 were neutered males, and 2 were intact males. Topical steroids were previously administered in 5 (23.8%) eyes; oral steroids were previously administered in 7 cats (36.8% of patients); three patients received both oral and topical steroids. Thirteen of 21 (61.9%) eyes had a history of ocular disease including ulcerative and nonulcerative keratitis, anterior uveitis, corneal sequestrum, conjunctivitis, and glaucoma. Median duration of NMF was 15 days (range 6-30 days). Follow-up ranged from 12 to 1601 days (median 169 days). Corneal perforation occurred in 1 (4.7%) eye and was successfully repaired. One lesion (4.7%) in a diabetic patient did not resolve. Nineteen of the treated eyes (90.5%) resolved with no complications. CONCLUSIONS: A nictitating membrane flap successfully treated 90.5% of FACH eyes (89.5% of patients).

Immunogenicity in dogs and protection against visceral leishmaniasis induced by a 14kDa Leishmania infantum recombinant polypeptide.

In areas were human visceral leishmaniasis (VL) is endemic, the domestic dog is the main parasite reservoir in the infectious cycle of Leishmania infantum. Development of prophylactic strategies to lower the parasite burden in dogs would reduce sand fly transmission thus lowering the incidence of zoonotic VL. Here we demonstrate that vaccination of dogs with a recombinant 14kDa polypeptide of L. infantum nuclear transport factor 2 (Li-ntf2) mixed with adjuvant BpMPLA-SE resulted in the production of specific anti-Li-ntf2 IgG antibodies as well as IFN-γ release by the animals' peripheral blood mononuclear cells stimulated with the antigen. In addition, immunization with this single and small 14kDa poplypeptide resulted in protracted progression of the infection of the animals after challenging with a high dose of virulent L. infantum. Five months after challenge the parasite load was lower in the bone marrow of immunized dogs compared to non-immunized animals. The antibody response to K39, a marker of active VL, at ten months after challenge was strong and significantly higher in the control dogs than in vaccinated animals. At the study termination vaccinated animals showed significantly more liver granulomas and lymphoid hyperplasia than non-vaccinated animals, which are both histological markers of resistance to infection. Together, these results indicate that the 14kDa polypeptide is an attractive protective molecule that can be easily incorporated in a leishmanial polyprotein vaccine candidate to augment/complement the overall protective efficacy of the final product.

Definition, Classification, and Pathophysiology of Canine Glaucoma.

Glaucoma is a common ocular condition in humans and dogs leading to optic nerve degeneration and irreversible blindness. Primary glaucoma is a group of spontaneous heterogeneous diseases. Multiple factors are involved in its pathogenesis and these factors vary across human ethnic groups and canine breeds, so the clinical phenotypes are numerous and their classification can be challenging and remain superficial. Aging and oxidative stress are major triggers for the manifestation of disease. Multiple, intertwined inflammatory and biochemical cascades eventually alter cellular and extracellular physiology in the optic nerve and trabecular meshwork and lead to vision loss.

Microscopic Lesions in Canine Eyes with Primary Glaucoma.

Although the clinical classification of primary glaucoma in dogs is quite simple, the phenotypes of glaucoma in most of the species are indeed multiple. Ophthalmologists can often evaluate the dynamic changes of clinical signs at different times in the course of the disease, whereas pathologists are often presented with globes that have undergone abundant therapies and are at the end stage. Therefore, an open collaboration between clinicians and pathologists can produce the most accurate interpretation in the pathology report and improve patient outcomes. This article focuses on the histomorphologic elements that characterize, and are important to, canine primary glaucomas.

Functional Anatomy of the Outflow Facilities.

In order to understand the pathophysiology, select optimal therapeutic options for patients and provide clients with honest expectations for cases of canine glaucoma, clinicians should be familiar with a rational understanding of the functional anatomy of the ocular structures involved in this group of diseases. The topographical extension and the structural and humoral complexity of the regions involved with the production and the outflow of aqueous humor undergo numerous changes with aging and disease. Therefore, the anatomy relative to the fluid dynamics of aqueous has become a pivotal yet flexible concept to interpret the different phenotypes of glaucoma.

Medical Treatment of Primary Canine Glaucoma.

Glaucoma is a painful and often blinding group of ocular diseases for which there is no cure. Although the definition of glaucoma is rapidly evolving, elevated intraocular pressure (IOP) remains the most consistent risk factor of glaucoma in the canine patient. Therapy should be aimed at neuroprotection. The mainstay of therapy focuses on reducing IOP and maintaining a visual and comfortable eye. This article discusses the most current ocular hypotensive agents, focusing on their basic pharmacology, efficacy at lowering IOP, and recommended use in the treatment of idiopathic canine glaucoma.

Mast cell tumor invading the cornea in a horse.

A 3-year-old Marwari mare was presented for evaluation of an irregular, reddish mass protruding from behind the right third eyelid. The mass appeared to arise at the ventral limbal area, involved the perilimbal bulbar conjunctiva and widely extended into corneal tissue. No other ocular or systemic abnormalities were detected at the time of presentation. The mass was surgically removed by lamellar keratectomy, with defocused CO(2) laser used as adjunctive therapy to treat the surgical exposed area and its surroundings. Histopathologic evaluation showed sheets of densely packed, well-differentiated neoplastic mast cells separated by fibrovascular connective tissue. Nuclear staining for Ki-67 was performed, and an average of 370 cells were positive per 1000 counted cells. Two months postoperatively, the surgical site was filled with flat fibrovascular and pigmented tissue, while the surrounding cornea was transparent with no superficial vascularization around the fibrotic scar. Thirty-two months after treatment, no recurrence of the neoplasia was reported.

Corneal sequestrum in a dog with chronic unilateral keratoconjunctivitis sicca.

CASE DESCRIPTION: A 14-year-old 8.2-kg (18.04-lb) castrated male Cairn Terrier with chronic keratoconjunctivitis sicca in the left eye was evaluated because of severe blepharospasm and a black plaque of 3 weeks' duration. CLINICAL FINDINGS: Abnormalities of the left eye included a decreased Schirmer tear test value and the presence of a brownish-black plaque in the center of the cornea. The plaque was surrounded by fibrovascular tissue except at the medial aspect where there was mild malacia of the adjacent corneal stroma. TREATMENT AND OUTCOME: The plaque was removed by superficial keratectomy, and a conjunctival graft was performed. Histologic evaluation of the plaque and surrounding cornea revealed ulceration, stromal necrosis, and chronic suppurative keratitis with fibrosis and neovascularization. Evaluation of plaque sections that were stained with Gram and Von-Kossa stains yielded negative results for bacteria and mineralization, respectively; examination of sections stained with periodic acid-Schiff stain revealed multiple intracytoplasmic inclusions in macrophages. Virus isolation and a PCR assay for canine herpesvirus yielded negative results. Transmission electron microscopy revealed collagen disruption with interspersed macrophages and apoptotic keratocytes; no viral particles or evidence of other infectious agents was observed. The graft healed without complication and was trimmed 2 weeks after surgery. Four months after surgery, the Schirmer tear test value remained decreased from reference limits despite topical tacrolimus treatment, and pigmentary keratopathy was present surrounding the graft. CLINICAL RELEVANCE: Corneal sequestra are rare in species other than cats. In this dog, it was possible that chronic keratoconjunctivitis sicca might have contributed to the development of the corneal sequestrum.

A technique for evisceration as an alternative to enucleation in birds of prey: 19 cases.

Ocular trauma is common in birds of prey presented to wildlife clinics and rehabilitation centers. Enucleation is the procedure most commonly described for treatment of end-stage ocular disease or chronically painful eyes in birds; however, there are several disadvantages and risks to this procedure. While evisceration has been suggested as an alternative, it has not been described for multiple cases or with long-term follow-up data in birds of prey. This report details an evisceration technique performed in 5 captive birds of prey of 4 different species (1 eastern screech owl [Megascops asio], 1 great horned owl [Bubo virginianus], 2 red-tailed hawks [Buteo jamaicensis], and 1 bald eagle [Haliaeetus leucocephalus]) with long-term follow-up information. In addition, this report describes 14 cases of free-living owls of 3 different species (1 great horned owl, 4 barred owls [Strix varia], and 9 eastern screech owls) on which this technique was performed from 2004 to 2011 and which were subsequently released to the wild. Because of the limited risk of complications and the less-severe disruption of facial symmetry, which may be particularly important in owls that are candidates for release to the wild, evisceration should be considered over enucleation in birds of prey that require surgical intervention for the management of severe sequelae to ocular trauma.

Western blot patterns of serum autoantibodies against optic nerve antigens in dogs with goniodysgenesis-related glaucoma.

OBJECTIVE: To investigate whether differences existed between clinically normal dogs and dogs with goniodysgenesis-related glaucoma (GDRG) in serum autoantibodies against optic nerve antigens. ANIMALS: 16 dogs with GDRG, 17 healthy dogs with unremarkable pectinate ligament and iridocorneal angle morphology, and 13 euthanized dogs with no major ocular abnormalities or underlying diseases. PROCEDURES: Western blotting was performed with optic nerve extracts from the euthanized dogs as an antigen source and serum from clinically normal dogs and dogs with GDRG as a primary antibody (autoantibody) source. Blots were evaluated for presence and density of bands. RESULTS: Multiple bands were identified on western blots from all dogs with GDRG and all clinically normal dogs, with a high degree of variability among individual dogs. Dogs with GDRG were significantly more likely than healthy dogs to have bands present at 38, 40, and 68 kDa. Dogs with GDRG had significant increases in autoreactivity at 40 and 53 kDa and a significant decrease in autoreactivity at 48 kDa. CONCLUSIONS AND CLINICAL RELEVANCE: Significant differences in serum autoantibodies against optic nerve antigens were found in dogs with versus without GDRG. Although it remains unclear whether these differences were part of the pathogenesis of disease or were sequelae to glaucomatous changes, these findings provide support for the hypothesis that immune-mediated mechanisms play a role in the development or progression of GDRG. However, the high degree of variability among individual dogs and the considerable overlap between groups suggest that the clinical usefulness of this technique for distinguishing dogs with GDRG from clinically normal dogs is likely limited.

Surgical treatment of epibulbar melanocytomas by complete excision and homologous corneoscleral grafting in dogs: 11 cases.

OBJECTIVE: The aim of the study was to evaluate the efficacy, outcome, and complications following penetrating sclerokeratoplasty and frozen homologous corneoscleral grafting for the management of extensive canine epibulbar melanocytomas. METHODS: Medical records of canine patients treated at three different veterinary hospitals between 1999 and 2010 were reviewed. Signalment, location and extent of melanocytomas, recurrence rate, and early and late complications were reported. Patients were re-examined postoperatively to provide follow-up information. RESULTS: Patients included one intact male, three castrated males, six intact females, and one spayed female, with a median age of 5 years (range, 3-9). German Shepherds were overrepresented. Tumors extended from 2 to 4 clock hours at the limbus and up to 17 mm from clear cornea to globe equator. One case showed iridocorneal angle invasion; corneal involvement was present in all cases, and lipid keratopathy was present in four cases. In two cases, there was incomplete resection owing to tumor extent. Follow-up time ranged from 3 to 72 months (median, 17 months), with one case of intraocular tumor progression. Early complications included anterior uveitis (11/11), intracameral fibrin (5/11), hyphema (4/11), corneal edema (4/11), exuberant corneal granulation tissue (2/11), focal retinal edema (1/11), dyscoria (1/11), and partial suture dehiscence (1/11). Late complications included corneal fibrosis and/or pigmentation (11/11), faint anterior cortical cataracts (3/11), and lipid keratopathy (1/11). Vision was retained in all cases. CONCLUSIONS: This technique offers a surgically challenging but effective treatment for extensive epibulbar melanocytomas. In this case series, complications were mild and transient, with preservation of ocular anatomy and function.

Histopathologic evaluation of the anterior segment of eyes enucleated due to glaucoma secondary to primary lens displacement in 13 canine globes.

PURPOSE To describe histologic anterior segment changes in eyes affected with primary lens displacement (PLD) and secondary glaucoma. METHODS Histologic sections stained with H&E from canine eyes enucleated because of PLD and secondary glaucoma were examined. RESULTS Thirteen eyes from 12 patients were evaluated. Four dogs were castrated males and eight spayed females. Median age was 8 years of age (range 3-13). Breeds included seven terriers and five other breeds. All eyes examined demonstrated varying degrees of inflammation involving the iris and cleft. Mononuclear and melanophagic infiltration of the cleft was found in all specimens. Four globes also showed polymorphonuclear infiltrate. Pre-iridal fibrovascular membranes were clearly identified in 10 of 13 eyes. Total inflammatory score was significantly greater in all globes examined compared with an age-matched group of normal dogs. The posterior pigmented iris epithelium demonstrated a consistent pattern of hyperplasia and/or hypertrophy and cystic degeneration, more prominent in the more central regions. In some cases, hyperplasia was of greatest severity in the mid-iris and associated with thinning or flattening of the pupillary region. CONCLUSIONS These results suggest that lens instability may be associated with chronic inflammation and secondary glaucoma. Mechanical irritation from an unstable lens may result in hypertrophy and/or hyperplasia of the posterior pigmented iris epithelium and subsequent cellular exfoliation and release of melanin. An inflammatory reaction directly or indirectly related to melanin release may obstruct the outflow pathways ultimately leading to glaucoma and loss of vision. Use of topical steroids may be warranted in dogs with PLD.

Anterior segment fluorescein angiography of the normal feline eye using a dSLR camera adaptor.

PURPOSE: To describe anterior segment fluorescein angiography (ASFA) of the normal feline eye using a digital single-lens reflex (dSLR) camera adaptor. ANIMALS: Ten cats free of ocular and systemic disease were evaluated. METHODS: All cats received maropitant citrate (1.0 mg/kg SQ) and diphenhydramine (2.0 mg/kg SQ) 20 min prior to anesthesia using propofol (4 mg/kg IV bolus, 0.2 mg/kg/min CRI). Standard color and red-free images were obtained prior to the administration of 10% sodium fluorescein (20 mg/kg IV). Imaging was performed using a dSLR camera (Canon 7D), dSLR camera adaptor, camera lens (Canon EF-S 60 mm f/2.8 macro), and an accessory flash (Canon 580EXII). Imaging occurred at a rate of 1/second immediately following IV bolus of sodium fluorescein for a total of 30 s, then at 1, 2, 3, 4, 5, and 10 min. RESULTS: Ten cats with an average age of 3.7 ± 0.9 years and various iris colors were imaged. Arterial, capillary, and venous phases occurred 4.6, 7.8, and 8.9 s postinjection, respectively. Visibility of the vasculature was not impaired by the degree of iris pigmentation. Patency of a persistent pupillary membrane was noted in one cat. Vessel leakage was common, as well as, leakage into the aqueous humor. Proper patient positioning and restricted ocular movements were critical. No adverse events were noted. CONCLUSIONS: This study demonstrates ASFA findings in normal feline eyes using a cost-effective dSLR camera adaptor. Fluorescein leakage from vessels and into the aqueous humor was a common finding. Visibility of iris vasculature was not impaired by the degree of iris pigmentation.

Glaucoma associated with uveal cysts and goniodysgenesis in American Bulldogs: a case series.

We present a series of three American Bulldogs with clinical signs of glaucoma and intraocular inflammation accompanied by bilateral uveal cysts and abnormal gonioscopic findings. All dogs proved refractory to medical management and were enucleated. Histopathologic findings were similar in all three and included significant preiridal fibrovascular membranes and mononuclear inflammatory infiltrates in the anterior uvea. On microscopic evaluation, cysts appeared to arise primarily from the ciliary body and iridociliary sulcus, with smaller cysts also budding from the posterior iris. Pigment dispersion was variable but consistent, involving deposition of a small number of pigment-laden cells in the dependent trabecular meshwork. Cataract formation was not noted. Glaucoma associated with uveal cysts has been described previously in Golden Retrievers and Great Danes, although clinical and histopathologic findings in those breeds are not identical to those described here. American Bulldogs with uveal cysts should have gonioscopy performed and should be monitored carefully for signs of increased intraocular pressure and intraocular inflammation. Furthermore, documentation of cyst-associated glaucoma in a third breed suggests clinicians should exercise caution in dismissing uveal cysts in dogs as incidental findings.