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Veterinary Ophthalmology (2009) 12, 6, 390–409 Abstracts: 40th Annual Meeting of the American College of Veterinary Ophthalmologists, Chicago, IL, USA November 4–7, 2009 ABSTRACT NO.: 001 Establishing a reproducible method for the culture of primary equine corneal cells R. L. Mathes,* U. M. Dietrich,* T. M. Krunkosky,† D. J. Hurley‡ and A. J. Reber‡ *Department of Small Animal Medicine and Surgery, College of Veterinary Medicine, University of Georgia, Athens, GA, USA, †Department of Anatomy and Radiology, College of Veterinary Medicine, University of Georgia, Athens, GA, USA, ‡Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA, USA Purpose: To establish a reproducible method for the culture of primary equine corneal epithelial cells, keratocytes and endothelial cells and to describe each cell’s morphologic characteristics, immunocytochemical staining properties and conditions required for cryopreservation. Methods: Corneas from eight horses recently euthanized for reasons unrelated to this study were collected aseptically and enzymatically separated into three individual layers for cell isolation. The cells were plated, grown in culture and continued for several passages. Each cell type was characterized by morphology and immunocytochemical staining. Results: All three equine corneal cell types were successfully grown in culture. Cultured corneal endothelial cells were large, hexagonal cells with a moderate growth rate. Keratocytes were small, spindloid cells that grew rapidly. Epithelial cells had heterogenous morphology and grew slowly. The endothelial cells and keratocytes stained positive for vimentin and were morphologically distinguishable from one another. The epithelial cells stained positive for cytokeratin. Keratocytes and endothelial cells were able to be cryopreserved and recovered. The cryopreserved cells maintained their morphological and immunocytochemical features after cryopreservation and recovery. Conclusion: This work establishes reproducible methods for isolation and culture of all three equine corneal cells. Cell morphology and cytoskeletal element expression for each cell type is also described. This has not previously been reported for equine corneal cells. This report also demonstrates the ability to preserve equine keratocytes and endothelial cells for extended periods of time and utilize them long after primary cell collection, a feature that has not been reported for veterinary corneal cell culture. University of Georgia Veterinary Ophthalmology Research Fund (VORF). ABSTRACT NO.: 002 Serum antibodies against alpha A- and B-crystallin in American Cocker Spaniels N. Kanemaki,* Y. Kondo,* Y. Ichikawa* and D. E. Brooks† *Veterinary Teaching Hospital, Azabu University, Japan, †College of Veterinary Medicine, University of Florida, Gainesville, FL, USA Purpose: To evaluate serum antibody levels against alpha A- and B-crystallins in American Cocker Spaniel dogs (ACSs). Methods: Frozen lenses were homogenized, and water-soluble lens protein collected. The alpha A- and B-crystallins were separated with gel electrophoresis and evaluated by Western immunoblot analysis. Results: Seventy-one ACSs were evaluated including 13 dogs with hypermature cataracts, 27 dogs with mature cataracts, 2 dogs with immature cataracts, 5 dogs with IOL implantation, 6 glaucomatous dogs, 4 dogs with retinal disease, 5 others with ocular surface abnormalities, and 9 normal ACS dogs. The incidence of serum antibody against alpha A-crystallin (41.9%) in ACSs with eye diseases was higher than that of antibody against alpha B-crystallin (4.8%) in dogs with eye problems. In normal dogs the serum antibody against alpha A-crystallin (55.6%) was higher than that of antibody against alpha B-crystallin (22.2%). Serum antibodies against alpha A-crystallins were detected in dogs with various categories of eye disease. The serum antibody against alpha A-crystallin in cataractous dogs was relatively low compared to that in normal dogs. The intensity of alpha A-crystallin antibody titers in hypermature cataractous dogs associated with uveitis was significantly lower than in cases without uveitis. Conclusions: The formation of lens-crystallin antibodies is lower in normal ACS, and varies with the degree of cataract maturation in ACS. ABSTRACT NO.: 003 Vascular endothelial growth factor (VEGF) in diabetic and non-diabetic canine cataract patients K. L. Abrams,* P. F. Stabila,† K. Kauper† and S. Elliot† *Veterinary Ophthalmology Services, Inc. Warwick, RI, USA, †Neurotech USA, Inc., Lincoln, RI, USA Purpose: To measure vascular endothelial growth factor (VEGF) levels in aqueous humor, serum, and plasma in diabetic and non-diabetic cataractous dogs. Methods: Canine VEGF was assayed in the plasma and serum of 32 dogs (20 diabetics; 12 non-diabetics) and aqueous humor in 57 eyes of those dogs (39 diabetic; 18 non-diabetic) undergoing phacoemulsification, using a commercial canine VEGF assay. Statistical analysis was performed using Fisher’s PLSD, t-test, and regression analysis to compare values by diabetic status, duration of diabetes, age, weight, gender, left vs. right eye, and blood condition. Results: Plasma, but not serum or aqueous VEGF values of diabetics were significantly greater than non-diabetics (P = 0.019). Older nondiabetics (10–15 years) had higher VEGF values than younger (0–5 and 5–10 years) dogs (P = 0.0002 and 0.0001, respectively). There was a high correlation of aqueous VEGF between left and right eyes in all patients. Serum and plasma VEGF values in females were significantly higher than males in both groups. Conclusion: Similar to human diabetic patients, VEGF values in canine diabetic plasma is significantly greater than non-diabetics. However, aqueous humor values of VEGF in diabetic dogs is not greater than non-diabetics, whereas, aqueous VEGF values in human diabetics are elevated and correlate with the severity of diabetic retinopathy. ABSTRACT NO.: 004 Transfrontal orbitotomy in the dog N. Wallin-Hakansson and B. Wallin-Hakansson Referral Animal Hospital Stromsholm, Sweden Purpose: To describe an adaptable and extensive method for orbitotomy in the dog. Methods: A three step technique for orbitotomy was developed and applied in nine consecutive cases. The steps are zygomatic arch resection laterally, temporalis muscle elevation medially and zygomatic process osteotomy anteriorly-dorsally. The entire orbit is accessed with excellent exposure and latitude for surgical manipulation. Osteotomies are closed using cerclage wire. Soft tissues are sutured routinely. Facial nerve, lacrimal nerve and lacrimal gland functions are preserved. The procedure can easily be converted into an orbital exenteration. Results: Access to the orbit was sufficient in all cases. Anatomically correct closure was achieved. Complications included moderate, reversible swelling in two cases and mild in seven. Wound infection or emphysema did not occur. Postoperative discomfort appeared limited. No other complications attributable to the operative procedure occurred. Blinking ability and lacrimal function as determined by Schirmer tear test I were preserved. Follow-up time ranged from 12 months to 4 years. Conclusions: Transfrontal orbitotomy in the dog offers excellent exposure and latitude for manipulation. Complications are mild and temporary, anatomically correct closure is easy and postoperative discomfort limited. No funding and grants. ABSTRACT NO.: 005 Local anesthetic retrobulbar block reduces postoperative pain following enucleation of canine eyes: a randomized, placebo controlled, double-masked clinical trial K. E. Myrna, E. Bentley and L. J. Smith Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI, USA Purpose: To assess the efficacy of retrobulbar bupivicaine nerve block on postoperative analgesia following enucleation of canine eyes. Methods: Client-owned dogs admitted to the veterinary medical teaching hospital for routine enucleation were enrolled with owner consent and randomly assigned to a treatment (bupivicaine) or placebo (saline) group. Baseline subjective pain scores were recorded. Anesthesia consisted of hydromorphone and midazolam preoperatively, thiopental or propofol for induction and isofluorane in oxygen for maintenance. An inferior-temporal palpebral retrobulbar injection of either saline or bupivicaine was administered. Transpalpebral enucleation surgery was performed. Pain scores were recorded at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h after t = 0 (extubation) by observers masked to treatment groups. Rescue hydromorphone (0.2 mg kg IM or IV) was administered if subjective pain score totaled ‡9 or ‡3 in any one category. Incidence of treatment failure was analyzed using Fisher’s exact test. Significance was defined as P < 0.05. Results: Incidence of treatment failure was 72% (8/11) in saline-treated dogs and 9% (1/11) in bupivicaine-treated dogs (P < 0.0075). Time-totreatment failure (TTF) averaged 0.53 h among the nine failure dogs with the shortest TTF being 0.25 h and the longest 2 h. Conclusions: The study suggests that retrobulbar administration of bupivicaine in dogs, with traditional premedication, is an effective form of adjunctive analgesia and significantly reduces the need for additional postoperative analgesics after enucleation surgery. Funded by a grant from the American College of Veterinary Ophthalmologists Vision for Animals Foundation ABSTRACT NO.: 006 Cellular localization of Visudyne as a function of time after local injection for photodynamic therapy: an investigation into the mechanisms of tumor cell death L. D. Barnes, E. A. Giuliano and J. Ota Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri, Columbia, MO, USA Purpose: To evaluate time effects on cellular localization of Visudyne after local injection. Methods: A squamous cell carcinoma (SCC) cell line (A-431) was injected into right and left dorsolumbar subcutaneous tissue of mice. In experiment 1 (Exp 1) and 2 (Exp 2), the treatment (T) tumors received a local injection of Visudyn (Novartis, Priority Health Care, FL) (0.1 mg/cm3), and control (C) tumors received an equal dose of 5% dextrose in water (D5W). Mice were randomly subdivided into two groups (A & B; n = 5/group). Mice in Exp 1A and B were sacrificed 1 minute (min) and 30 minutes (mins) after local injection, respectively. Experiment 1 tumors were evaluated by fluorescence microscopy to determine drug localization. Experiment 2A and B tumors were laser irradiated 1 and 30 mins after injection, respectively and evaluated by transmission electron microscopy (TEM) to determine ultrastructural tumor cell damage. Results: Fluorescence was detected within the cytoplasm of T tumors in both Exp 1A and B. Significance was detected in fluorescence intensity between T1 min vs. T30 mins (P = 0.03) and between T1 min and C1 min tumors (P = 0.01), respectively. Tumors in Exp 2 demonstrated signs of apoptosis. Conclusions: Fluorescence microscopy demonstrated higher Visudyne concentration within SCC cytoplasm of 1 min compared to 30 min tumors. Transmission electron microscopy results revealed that tumors treated by photodynamic therapy (PDT) within 30 mins of local injection undergo cellular apoptosis. Supported by the ACVO Vision for Animals Foundation Grant. 2009 American College of Veterinary Ophthalmologists abstracts 391 ABSTRACT NO.: 007 Safety assessment of intracapsular moxifloxacin in canine eyes undergoing lens replacement surgery J. C. Wolfer Islington Animal Clinic, Toronto, ON, Canada Purpose: To evaluate the safety of intracapsular moxifloxacin HCl 0.5% (Vigamox) in canine eyes undergoing lens replacement surgery. Method: A total of 100 lg of moxifloxacin HCl (Vigamox) was injected into the lens capsule at the end of cataract surgery in 20 dogs, 29 eyes. The first two patients acted as controls, with each dog receiving the testcompound in one eye and BSS in the other. Patients were evaluated for any complications such as rise in intraocular pressure (IOP), corneal edema, uveitis, glaucoma, fibrin formation or chorioretinitis, on postoperative days 1, 7, 14, 30 and 90. Patient evaluation consisted of slit-lamp biomicroscopy, indirect ophthalmoscopy, fluorescein dye test and intraocular pressure measurement. Results: In all eyes treated there were no complications that were attributed to the use of intracapsularmoxifloxacin. Conclusion: Intracapsular moxifloxacin HCl, at a dose of 100 lg, is safein the canine eye. ABSTRACT NO.: 008 Expression of TLR-2, TLR-4, TRAP and IL-6 genes in normal canine eyes and in an eye with idiopathic uveitis, a pilot study J. Ota-Kuroki,* A. M. Stoker,† H. J. Sims,† C. P. Moore,‡ J. W. Pearce,‡ D. Buss‡ and K. Kuroki† *Midwest Veterinary Referral Center, Chesterfield, MO, USA, †Comparative Orthopaedic Laboratory, University of Missouri-Columbia, Columbia, MO, USA, ‡Department of Veterinary Medicine and Surgery, University of Missouri-Columbia, Columbia, MO, USA Purpose: To investigate the expression of toll-like receptor-2 (TLR-2), TLR-4, tartrate-resistant acid phosphatase (TRAP) and interleukin-6 (IL-6) in the uvea in normal canine eyes and in an eye with idiopathic uveitis. Methods: Anterior uveal tissue was collected from 14 normal eyes of 7 dogs and one eye with canine idiopathic uveitis. Real time polymerase chain reaction (PCR) was performed to evaluate TLR-2, TLR-4, TRAP and IL-6 gene expression level. In order to confirm TLR-4 expression at the protein level, TLR-4 immunofluorescence was also evaluated in all tissues. Results: TLR-2, TLR-4, TRAP and IL-6 gene expressions were detected by RT-PCR in all anterior uveal tissues examined. Expression levels of TLR-2, TLR4, TRAP, and IL-6 in the anterior uveal tissue was 6.6, 3.9, 18.5 and 2.3 times higher respectively in the eye with uveitis compared to the normal eyes. TLR-4 immunofluorescence was observed in all normal uveal tissues examined and also in the eye with idiopathic uveitis. Stronger TLR-4 immunostaining was observed in the case of idiopathic uveitis. Conclusion: This study demonstrated the presence of TLR-2, TLR-4, TRAP and IL-6 expression in normal canine anterior uveal tissues. Expression of these genes in the anterior uveal tissue appears to be increased in canine idiopathic uveitis compared to normal controls. Up-regulation of these molecules might play a significant role in disease initiation and progression of canine idiopathic uveitis and warrants further evaluation and case recruitment. ABSTRACT NO.: 009 Efficient gene delivery in the equine cornea D. G. Buss,* A. Sharma,† E. A. Giuliano* and R. R. Mohan*,† *College of Veterinary Medicine, University of Missouri-Columbia, Columbia, MO, USA, †Harry S. Truman Memorial Veterans Hospital, Columbia, MO, USA Purpose: We hypothesize that AAV5 vector can efficiently deliver a marker gene into the equine cornea without causing adverse cellular side effects. The aims of this study were to: (i) evaluate efficacy of AAV5 expressing EGFP marker gene for equine corneal fibroblasts (ECF) and (ii) establish the safety of AAV5 vector for equine corneal gene therapy. Methods: Cultured ECF were used from healthy harvested equine corneas. Cultures were maintained at 37C in humidified atmosphere with 5% CO2. AAV serotype 5 expressing EGFP under control of hybrid CMV + CBA promoters were applied topically to ECF tissue cultures. Fluorescence was observed in ECF cells expressing EGFP. Using DAPI staining the total number of cells were determined and compared to the number of fluorescent cells thereby establishing the transduction efficiency. Cellular apoptosis was measured using TUNEL assay to determine AAV safety. Results: Transduction of ECF cells with topically applied AAV5 expressing EGFP was successful. A transduction rate of 13.1% was found. No significant cell death was detected using TUNEL assay. Conclusions: Topically applied AAV5 vector is an efficient and safe option for gene therapy in in-vitro equine cornea. Further studies using AAV5 to determine the effects of various genes on the equine cornea are warranted. Gene therapy may be a useful therapeutic strategy for equine corneal disease. Supported by the University of Missouri Clinician Scientist Award. ABSTRACT NO.: 010 Bilateral optic atrophy in cynomolgus macaques used in toxicologic research R. R. Dubielzig,* R. Leedle,† T. M. Nork,* J. N. VerHoeve* and B. J. Christian† *Comparative Ophthalmic Research Laboratories, University of Wisconsin, Madison, WI, USA, †Covance Laboratories Inc, Madison, WI, USA, Purpose: The funduscopic findings, ERG, Flash VEP, and histopathologic findings of bilateral optic atrophy found in cynomolgus monkeys used in regulatory toxicologic research are described. Methods: Cynomologus macaques used for toxicologic studies had backgound lesions typical of bilateral optic atrophy, a condition previously described in Rhesus macaques. Fundus photography, ERG, and Flash VEP available in some of the monkeys and histopathology was performed in all animals. Results: The affected monkeys were 2–3 years old and all were from Vietnamese origin sources. In various studies between 0% and 24% of animals were affected. In one study with 14 affected animals, the histological appearance exhibited the following range of severity: severe (6), moderate (3), slight (3) and minimal (2). 4/6 with severe disease and 1/3 with slight disease had optic nerve atrophy detected on the pre-study fundus photography; all other animals were considered within the limits. Affected animals were not detected by ERG or Flash VEP in these animals. The morphologic changes consist of a decrease in ganglion cells in the macula and a decrease in axons in the temporal nerve fiber layer and the temporal optic nerve. The condition was always unrelated to the administration of test compounds. No affected animals had any detectable behavioral abnormalities. Conclusions: Bilateral optic atrophy has been reported in Rhesus macaques originating from China. We report the occurrence of a similar syndrome in cynomolgus monkeys of Vietnamese origin. Given the cynomolgus monkey is commonly used in ophthalmic drug safety studies, it is important to charaterize the nature of the finding. While the abnormality was not detected by ERG or flash VEP, the majority of markedly affected animals could be identified by careful fundus evaluation. ABSTRACT NO.: 011 Long-term control of equine recurrent uveitis using a suprachoroidal cyclosporine matrix reservoir drug delivery device B. C. Gilger,* J. H. Salmon,* D. A. Wilkie,† M. Utter,‡ A. Komaromy,‡ D. E. Brooks,§ R. Stoppini,– E. C. Ledbetter,** B. Nell†† and A. Labelle‡‡ *North Carolina State University, Raleigh, NC, USA, †The Ohio State University, Columbus, OH, USA, ‡New Bolton Center, University of Pennsylvania, PA, USA, §University of Florida, Gainesville, FL, USA, –Brescia, Italy, **Cornell University, Ithaca, NY, USA, ††Veterinary University of Vienna, Austria, ‡‡University of Illinois, Urbana, IL, USA Purpose: Recurrent uveitis (ERU) in the most common cause of blindness in horses. ERU is a T-cell mediated disease that is well controlled with cyclosporine (CsA) (Gilger et al, IOVS 2006; 47:2596–2605). The purpose of this study was to determine the long-term efficacy and duration of effect of a single, suprachoroidal matrix reservoir implant releasing CsA. Methods: Horses with ERU were implanted with a 6 mm diameter, 25 mg, reservoir matrix CsA implant in the deep sclera adjacent to the suprachoroidal space. Horses with follow-up greater than 1 year were examined for frequency of flares, long-term complications, and visual status at last recheck. Results: Data from 186 eyes of 156 horses that had CsA devices implanted for ERU was reviewed. Horses were from six centers in the US and two in Europe. Mean follow-up was 29 months (1–7 years). Horses with implants had significantly fewer flares after surgery (mean 0.05 flares/ month) than prior to implantation. Overall, 79.9% were visual at the last follow-up time. Only three eyes required a repeat implant, all 4 years or longer after the first implant. Conclusions: The deep sclera/suprachoroidal placement of the CsA device resulted in excellent long-term control of ERU. Because flares did not increase after the theoretical depletion of the CsA (3.2 years), autoreactive T-cells may undergo anergy without subsequent recurrence of disease. Support: State of North Carolina, North Caroline Uveitis Research Fund. Commercial Relationship: Gilger BC (none); Salmon JH (none); Wilkie DA (none); Utter M (none); Komaromy A (none); Brooks D (none); Stoppini R(none); Ledbetter EC (none); Nell B (none); Labelle A (none). ABSTRACT NO.: 012 Comparison of the effects of topical administration of a fixed combination latanoprost and timolol to monotherapy with latanoprost or timolol on intraocular pressure, pupil size, and heart rate in normal dogs. L. N. Smith,* P. E. Miller† and L. M. Felchle* *Eye Care for Animals,†University of Wisconsin-Madison, School of Veterinary Medicine, Madison, WI, USA Purpose: To determine whether a combination of latanoprost 0.005%/timolol 0.5% (LAT) (Pfizer, NYC, NY) administered topically BID has any greater effects on intraocular pressure (IOP), pupil size (PS), and heart rate (HR) in normal dogs than do latanoprost 0.005% (Pfizer, Inc, NYC, NY) or timolol 0.5% (Falcon Pharmaceuticals Ltd, Fort Worth, TX) monotherapy Methods: Seventeen normal dogs were randomly assigned to the treatment group (9) or the saline group (8). Three phases were conducted: LAT, latanoprost monotherapy, and timolol monothera. Baseline values were established on day 1 of each phase. On days 2 through 5, drugs were administered topically BID to one randomly chosen eye of dogs in the treatment group. The saline group received saline drops OU BID. IOP, PS, and HR were measured at 0, 2, 4, 6, 8, and 9 h after dosing for both groups. Results: At 2 through 9 h post-dose on days 2 and 5, the LAT and latanoprost treated eyes showed a significant decrease in IOP (P < 0.05) and a significantly smaller PS (P < 0.05) compared to dogs receiving timolol monotherapy or saline. LAT and timolol alone both significantly lowered HR (P < 0.05) compared to latanoprost and saline groups on days 2 and 5. Conclusions: Topical administration of latanoprost alone is as effective at lowering IOP as LAT when administered on a BID dosing schedule in normal dogs. Timolol, either alone or in combination, appears to have little effect on IOP in normal dogs; but causes a reduction in HR. ABSTRACT NO.: 014 Complications and visual outcomes associated with surgical correction of lens in the horse D. E. Brooks,* B. C. Gilger,† C. E. Plummer,* K. P. Barrie,* J. D. Lavach‡ and L. G. Karpinski§ *University of Florida, Gainesville, FL, USA, †North Carolina State University, Raleigh, NC, USA, ‡Eye Clinic for Animals, Reno, NV, USA, §Pinecrest Veterinary Hospital, Miami, FL, USA Purpose: To describe clinical findings and visual outcome of surgical therapy for lens luxation/ subluxation in the horse. Methods: Retrospective medical records study of horses that had surgical correction of lens luxation/subluxation at the University of Florida Veterinary Medical Center, North Carolina State University, and Eye Care for Animals were reviewed. Data collected from the medical records included signalment, clinical descriptions of ocular lesions, and visual outcomes. Results: Phacoemulsification was performed on five horses (two Quarterhorse mares, one Thoroughbred gelding, one Quarterhorse gelding, and one Arabian mare) with lens luxation/subluxation. Intracapsular lens extraction was performed on four horses (one standardbred filly, one Icelandic pony mare, one Arabian gelding, and one Quarterhorse gelding) with lens luxation/subluxation. Surgical complications included incision dehiscence, 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 392 a b s t r a c t s expulsive choroidal hemorrhage, retinal detachment, and infectious endophthalmitis. Three eyes were quickly enucleated postoperatively, five eyes became rapidly phthisical, and one eye was still visual ten years postoperatively. Conclusions: Surgical removal of a luxated lens in a horse is a difficult endeavor with a high rate of severe intraoperative and postoperative complications, and generally poor visual outcomes. ABSTRACT NO.: 015 Descemet’s membrane fragmentation in recurrent ‘‘orange’’ deep stromal abscesses in the horse D. E. Brooks, D. P. Taylor, C. E. Plummer, K. P. Barrie and J. F. Roberts University of Florida, Gainesville, FL, USA Purpose: To describe the clinical and histologic findings in eight horses with recurrent ‘‘orange’’ deep stromal abscesses (DSA). Methods: Retrospective medical records study of horses afflicted with ‘‘orange’’ DSA that appeared following the apparent resolution of the original DSA with medical and/or surgical therapy. Data collected from the medical records included signalment, types of ocular lesions, type of therapy performed, length of followup, complications and visual outcomes. Results: Medical records of eight horses that had ‘‘orange’’ DSA at the University of Florida Veterinary Medical Center from 2004 to 2007 were reviewed. Six cases occurred following lamellar keratoplasty, and two cases following medical therapy of DSA. Histology of the corneas found many neutrophils, macrophages, lymphocytes, plasma cells and multinucleated giant cells, and multifocally fragmented Descemet’s membrane. Gram positive cocci were found in the tissue and anterior chamber of four eyes. Aspergillus was cultured from one case and fungi found in Descemets in four eyes. Enucleation was performed in four of the post surgical cases and one of the post medical therapy cases. Lamellar keratoplasty was successful in two cases and medical therapy in another case. Conclusions: Recurrent DSA do not appear to be related to traumatic micropuncture. Active ‘‘infection’’ was indicated by histology and clinical behavior. Endothelial cell/Descemet’s reaction to fungi appears prominent and suggests microendothelial micropuncture as part of the pathogenesis for recurrent DSA. ABSTRACT NO.: 016 Conjunctival lymphoma in the horse D. E. Brooks,* D. Wilkie,† C. E. Plummer,* K. P. Barrie,* A. Dwyer,‡ A. G. Matthews,§ B. C. Gilger,– D. Hendrix** and J. P. Pickett†† *University of Florida, Gainesville, FL, USA, †Ohio State University, Columbus, OH, USA, ‡Genesee Valley Equine Clinic, Scottsville, NY, USA, §McKenzie, Bryson and Marshall Veterinary Surgeons, Kilmarnock, Scotland, –North Carolina State University, Raleigh, NC, USA, **University of Tennessee, Knoxville, TN, USA, ††Virginia Tech University, Blacksburg, VA, USA Purpose: To describe clinical findings and visual outcome of medical and surgical therapy for conjunctival lymphoma in the horse. Methods: Retrospective medical records study of horses that had conjunctival lymphoma at the University of Florida Veterinary Medical Center, Ohio State University, NY, North Carolina State University, University of Tennessee, and Scotland were reviewed. Data collected from the medical records included signalment, clinical descriptions of ocular lesions, and treatment outcomes. Results: T cell rich B cell lymphoma of the bulbar, palpebral and nictitans conjunctiva was diagnosed in 20 horses. The average age at presentation was 14.0 ± 7.5 years. Lesions were nodular or diffuse. Treatment consisting of resection, chemotherapy, or combined therapy was considered good initially in 15/20 cases, although seven horses were euthanized with systemic disease signs within 10 months. Nine horses remained apparently tumor free at 4.6± 3.5 (range: 2–10) years. Conclusions: Longterm prognosis for these cases is generally good, unless diffuse disease was present at initial presentation. ABSTRACT NO.: 017 Equine subepithelial keratomycosis D. E. Brooks, C. E. Plummer, K. P. Barrie, S. E. Blackwood, C. M. Nunnery and G. Ben-Shlomo Department of Large Animal Clinical Sciences, University of Florida, Gainesville, FL, USA Purpose: To describe clinical findings in equine subepithelial keratomycosis (SEK). Methods: Retrospective medical records study of horses that had subepithelial keratomycosis at the University of Florida Veterinary Medical Center from 2007 to 2009 were reviewed. Data collected from the medical records included signalment, clinical descriptions of ocular lesions, and therapeutic outcomes. Results: Fourteen horses with unilateral SEK were diagnosed. They included three Quarterhorse geldings, two Morgan geldings, one Morgan mare, one Arabian mare, one Arabian gelding, one Thoroughbred gelding, one Thoroughbred mare, one Appaloosa mare, two warmblood geldings and one Holsteriner mare. Multifocal punctate subepithelial opacities were present in all eyes. Intermittent phases of weak fluorescein and/or Rose Bengal retention were present in seven eyes. Clinical signs of iridocyclitis were absent in all eyes. Cytologic confirmation of fungi was found in five cases, and Aspergillus cultured in one case. All eyes with SEK resolved with topical antifungal therapy. Conclusions: This is the first clinical report of nonulcerative subepithelial keratomycosis in the horse and may represent a preliminary phase of equine keratomycosis. ABSTRACT NO.: 018 Anesthetic recovery following corneal surgery in horses (2006–2008) M. E. Utter, L. M. Getman and K. Wegner New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA, USA Purpose: To evaluate the hypothesis suggested by prior research that horses undergoing ocular surgery are at increased risk of poor anesthetic recovery by comparing recovery scores from corneal versus non-ocular surgeries using a case control retrospective design. Method: Records from 58 corneal surgeries performed in 2006–2008 were matched to a control on patient age, anesthesia time, surgery time, recovery time, ASA status, surgery date and anesthetist. Corneal surgeries included conjunctival, corneal and amnion grafts. Emergency surgeries and fracture repairs were excluded from controls. Recovery scores from 1 to 5 were assigned to each case and compared using a Mann–Whitney U-test. Quality of match between cases and controls was evaluated on matching variables using a Student’s t-test. Values of P < 0.05 were considered significant. Results: There was no statistical difference between groups in mean recovery scores. There was no statistical difference between corneal surgery cases and controls in mean age, surgery time, anesthesia time, or recovery time, suggesting good matching between cases and controls. Conclusions: Horses recovering from general anesthesia following corneal surgery appear at no greater risk than horses recovering from non-ocular surgeries. ABSTRACT NO.: 019 Feline conjunctival mastocytosis: eight cases C. S. Schobert and R. R. Dubielzig School of Veterinary Medicine, University of Wisconsin, Madison, WI, USA Purpose: Conjunctival mastocytosis is a rare lesion in cats. We report on the morphology and clinical features of 10 submissions representing 8 cats. Also reported are the preliminary results on the behavior and clinical outcome of this lesion. Methods: Eight cases of feline conjunctival mastocytosis were selected from the COPLOW collection, which includes more than 6000 feline specimens. The authors examined H&E, Giemsa and Toluidine Blue stained sections cut from paraffin blocks. Immunohistochemistry for CD117 (C-KIT) was also used. We described the state of the epithelium, presence in the epithelium and substantia propria of edema, mast cells and eosinophils. Follow-up information was received from all by fax and/or phone. Results: Edema was observed in all cases. Mast cells and eosinophils were seen in the epithelium and substantia propria. Six of the eight cases had mast cells in both the epithelium and substantia propria, while two cases demonstrated mast cells in the epithelium only. Seven of the eight cases contained eosinophils in the epithelium and/or substantia propria or both. Four of the eight cases displayed a papillated epithelial proliferation. One of the cases contained ‘‘foamy’’ cells of unknown significance. CD117 was not useful in this study due to very inconsistent, weak staining of mast cells. Of the eight cats, three showed recurrence and three did not, while two were unknown. One of the cases reportedly had multiple flare-ups. In one case, mastocytosis was found in the bulbar conjunctiva after initial diagnosis in the nictitans. Conclusions: Of the six cases with complete follow-up, 50% showed local recurrence. Eosinophils mingled with mast cells in both the epithelium and substantia propria were seen in all cases. The finding of mast cells directly within the epithelium is a distinctive feature of this condition. We speculate an allergic or viral etiology. ABSTRACT NO.: 020 Arteriovenous hemangioma affecting the anterior surface of the eye and the oral cavity in a cat Scott R. Earnest,* Melanie Mineo† and Richard R. Dubielzig* *School of Veterinary Medicine, University of Wisconsin, Madison, WI, USA, †Upstate Veterinary Specialists, Latham, NY, USA Purpose: We document the case of a cat with a severely destructive anterior surface ocular mass and an oral mass both diagnosed as arteriovenous hemangioma – a rare tumor most often diagnosed in the skin. Methods: This case was submitted to the Comparative Ocular Pathology Laboratory of Wisconsin for diagnosis and interpretation. H&E, Alcian Blue-PAS, and Trichrome stains were performed. Immunohistochemistry for smooth muscle actin was performed. Results: A seven-year-old female cat presented because of a progressive ocular deformity in the left eye, which had progressed over a one month period. The cat was unable to close the affected eye. An oral mass on the rostral mandibular gingival was found at the time of enucleation and was removed also. Grossly the anterior segment of the globe was markedly distorted and effaced by a mottled tan to red mass affecting the limbus, half of the cornea and filling most of the anterior chamber. The remainder of the lobe was unaffected. Histologically, the mass was characterized by large interconnected vascular structures often surrounded by disorganized plump spindle cells resembling smooth muscle. This impression was confirmed by immunohistochemistry for smooth muscle actin. The histology of the gingival mass was identical. The diagnosis was ocular and oral arteriovenous hemangioma. Two other cases of arteriovenous hemangioma were also found in the COPLOW collection, both from the skin near the eye. Conclusions: Arteriovenous hemangioma is reported in dogs, cats, and humans. It is a rare benign tumor most often diagnosed on the skin. One case is reported from the tongue of a dog but this is the first case reported from the eye. ABSTRACT NO.: 021 UV-absorbing contact lenses mediate UV-induced damage to the anterior segment H. L. Chandler, K. S. Reuter, L. T. Sinnott and J. J. Nichols The Ohio State University, College of Optometry, Columbus, OH, USA Purpose: To determine the impact of ultraviolet (UV)-absorbing contact lenses (CL) on the prevention of UV-induced pathological changes in the cornea, aqueous humor, and crystalline lens (associated with cataract formation). Methods: Twelve rabbits were assigned to one of three treatment groups (n = 4 per treatment group) as follows: (1) UV-absorbing CL (Acuvue Oasys, absorbs 99% of UV-B and 90% of UV-A); (2) non-UV absorbing CL (Focus Night and Day, no UV absorption);(3) no CL. The contralateral eye was patched without a CL. Animals received UV-B (1.667 J/cm2) and UV-A (0.245 J/cm2) exposures daily for 5 days. Post-mortem tissues were examined as follows: in the cornea, expression of matrix-metalloproteinases (MMPs) was evaluated by zymography and apoptosis was evaluated by TUNEL and caspase-3 ELISA; ascorbate in the aqueous humor was evaluated by NMR spectroscopy; in the crystalline lens apoptosis was evaluated by TUNEL and caspase-3 ELISA. Nonparametric statistical tests were used to compare outcomes accounting for treatment group and UV exposure status. Results: For the cornea, there was a statistically significant increase in MMP-2 and -9, TUNEL positive cells and caspase-3 activity in the exposed non-UV absorbing CL group compared to the exposed UV-absorbing CL group (all P = 0.03). A statistically significant decrease in aqueous humor ascorbate was observed in the exposed non-UV absorbing CL group compared to the exposed UV-absorbing CL group (P = 0.03). For the crystalline lens, there was a statistically significant increase in caspase-3 activity in the exposed non-UV absorbing CL group compared to the exposed UV-absorbing CL group (P = 0.03). There was an increased number of TUNEL positive cells in both the exposed non-UV absorbing CL and non-CL groups; this was not statistically significant compared to the exposed UV-absorbing CL group. There were no statistically significant differences noted in any of the outcomes when comparing the exposed 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 abstracts 393 non-UV absorbing CL group and the contralateral control eyes. Conclusions: MMPs can be induced within the cornea by UV exposure and are associated with many pathologic inflammatory cascades. Ascorbate is a scavenger of free radicals in the aqueous humor and can decrease following UV exposure. DNA damage and apoptosis can be induced in both the cornea and lens by UV exposure. These data show that UV-blocking CL is capable of protecting the cornea, aqueous humor, and crystalline lens from these UV-induced pathological processes. Ocular protection from UV-exposure is warranted in a variety of species. Supported by Vistakon, Division of Johnson & Johnson Vision Care Inc. C. ABSTRACT NO.: 022 Effect of dexamethasone sodium phosphate 0.1% on intraocular pressure in normal dogs treated with latanoprost 0.005% M. A. Gilmour* and T. W. Lehenbauer† *Department of Veterinary Clinical Sciences, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK, USA, †School of Veterinary Medicine, Veterinary Medical Teaching and Research Center, University of California-Davis, Tulare, CA, USA Purpose: To determine if dexamethasone 0.1%, when administered with latanoprost 0.005%, decreases the hypotensive and miotic effects of latanoprost in normal dogs. Methods: Sixteen dogs were equally divided into two groups. Dogs in Group I received 1 drop latanoprost (Cardinal Health, Woodstock, IL) O.U. and 10 min later received 1 drop neomycin/polymyxin B/ dexamethasone (NPD) (Alcon Laboratories, Fort Worth, TX) O.D. (4 dogs) and O.S. (4 dogs). The control eye for each dog received 1 drop neomycin/polymyxin B/gramicidin (NPG) (Bausch & Lomb, Tampa, FL). Dogs in Group II received 1 drop NPD O.D. (4 dogs) and O.S. (4 dogs) and the control eye received 1 drop NPG. Ten minutes later both eyes received 1 drop latanoprost. Treatment started on Day 0 at 8:00 a.m. and ended on Day 2 at 8:00 p.m. Latanoprost was admininstered q 24 h at 8:00 a.m. The NPD and NPG were administered at 8:00 a.m., 2:00 p.m. and 8:00 p.m. Intraocular pressure (IOP) and pupil diameter were measured at 8:00 a.m., 10:00 a.m., 12:00 p.m., 2:00 p.m. and 8:00 p.m. on Day 0 through Day 3 at 7:00 a.m. Results: There was no significant difference in pupil diameter between control and dexamethasone treated eyes. There was a significant difference in IOP between the control and treated eyes, with an overall average of 11% increase in the treated eyes. Order of administration of the latanoprost and dexamethasone did not have a significant effect on the change in IOP. Conclusions: Concurrent treatment with dexamethasone 0.1% and latanoprost 0.005% does decrease the hypotensive effect of latanoprost in normal dogs. ABSTRACT NO.: 023 Long term success of Ahmed gonioimplants in dogs with glaucoma D. V. H. Hendrix, H. D. Westermeyer and D. A. Ward College of Veterinary Medicine, University of Tennessee, Knoxville, TN, USA Purpose: To evaluate the long term success and describe the complications associated with gonioimplants in dogs with primary glaucoma. Methods: Nine dogs that had received a commercially available gonioimplant (Ahmed Glaucoma Valve, New World Medical, Rancho Cucamonga, CA) implanted routinely between 2000 and 2008 were included in the study. All dogs were treated with mitomycin C in the conjunctival pocket intraoperatively. A temporalis muscle fascia graft (n = 8) or porcine intestinal submucosa (n = 1) was used to cover the implant tube as it exited the globe. All dogs were treated with tissue plasminogen activator immediately postoperatively. Additionally, all dogs received 1% prednisolone acetate once daily until failure of the implant. Intervention with 5-FU injections or additional surgery was done if intraocular pressures rose above 20 mmHg or an upward trend was noted. Results: Median age was 9 years. Fewer glaucoma medications were needed to control intraocular pressure after gonioimplant placement. Median time to implant failure was 13 months (range 5–40 months). Reasons for implant failure included implant extrusion due to conjunctival dehiscence or necrosis. Eventually IOP rose above 20 mmHg after surgery in 7/9 dogs (median time 326 days). Intervention was done in 4 dogs to maintain satisfactory intraocular pressure. At 12 months, 7/9 cases retained vision, and 5/9 retained vision throughout followup. Conclusions: The use of a gonioimplant alone as described in this study is an effective therapy in treatment of medically refractory primary glaucoma in dogs. ABSTRACT NO.: 024 Canine episcleral onchocerciasis in 18 dogs J. M. Dreyfus and R. R. Dubielzig University of Wisconsin, School of Veterinary Medicine, Department of Pathobiological Sciences, Madison, WI, USA Purpose: This retrospective study evaluated the breed, age, lesion location and geographic distribution in 18 dogs diagnosed with granulomatous episcleritis associated with onchocerca infestation from 1994 to 2009 at the Comparative Ocular Pathology Lab of Wisconsin (COPLOW). Methods: COPLOW, a mail-in diagnostic laboratory, maintains a searchable database of submissions dating from 1978. In 2005, Zarfoss et al. reported on two cases of ocular onchocerciasis diagnosed at the lab. Since that time, we have diagnosed an additional 16 cases identified via database search. The current study reviewed diagnostic slides and case submission. Results: In 13 cases, nematodes were definitively identified as Onchocerca sp. based on the following morphologic features: low, rounded, circumferential external ridges and internal striae. An additional five cases were consistent with onchocerciasis, based on clinical interpretation, geographic location, and eosinophilic, granulomatous inflammation in periocular tissues. Nematode parasites without morphologic features sufficient to positively identify its genus were present in one of the five presumptive diagnoses. Breeds of affected dogs include: Australian shepherd (3), Labrador Retriever (2), and one each of Bassett Hound, Blue heeler, Border Collie, Boxer, Catahoula, Chow mix, Dachshund cross, English Mastiff, Fox Terrier, German Shepherd, German Shepherd cross, Red Heeler cross, Rottweiler. The average age of 11 males and 7 females is 5 years ranging from 1 to 11. The laterality of the ocular tissue submitted was (7-OD/8-OS/1-OU). All but 2 of 18 cases originate from clinics in the metropolitan regions of Los Angeles, CA (8), San Diego, CA (5), and Phoenix, AZ (3). Single cases came from Utah and Nevada. Conclusion: These findings suggest that canine episcleral onchocerciasis is a continuing problem in very specific regional areas of the United States and that dog breeds that are likely to be out of doors are primarily affected. Financial conflict. ABSTRACT NO.: 025 Topical ocular corticosteroid administration in dogs with experimentally induced latent canine herpesvirus-1 infection N. C. Kice,* E. C. Ledbetter,* S. G. Kim,† R. B. Matusow* and E. J. Dubovi† *Cornell University, College of Veterinary Medicine, Department of Clinical Sciences, Ithaca, NY, USA, †Cornell University, College of Veterinary Medicine, Department of Population Medicine and Diagnostic Sciences, Ithaca, NY, USA Purpose: To determine if topical ocular corticosteroid administration reactivates latent canine herpesvirus-1 (CHV-1) in adult dogs. Methods: A randomized, masked, placebo-controlled, crossover study design was used. Ten adult specific pathogen-free beagles were included in the study. Primary ocular CHV-1 infection was experimentally induced in the dogs by ocular inoculation and the presence of reactivatable latency was later confirmed by administration of an immunosuppressive dosage of systemic corticosteroid. Twelve months following experimental CHV-1 reactivation, dogs were administered either topical ocular prednisolone acetate (1.0% ophthalmic suspension, one drop, OU, q6h) or placebo (artificial tear solution, one drop, OU, q6h) for 28 days. After a 14 day washout period, the treatment groups were reversed. Ophthalmic examinations, ocular sample real-time quantitative CHV-1 polymerase chain reaction assays, and CHV-1 serum neutralization (SN) titers were performed at regular intervals. The frequency of clinical ocular disease, ocular viral shedding, and 4-fold elevations in CHV-1 SN titers were statistically compared between treatment and control groups. Results: Viral reactivation was not detected more frequently in dogs administered topical ocular prednisolone than dogs administered placebo as determined by clinical ocular disease recrudescence, ocular viral shedding, and serologic response. Conclusions: This study suggests administration of topical ocular prednisolone with the evaluated drug concentration, dosing frequency, and treatment duration is not likely to result in detectable reactivation of latent CHV-1. Systemic absorption of locally administered corticosteroid may be insufficient to reactivate latent virus and produce recurrent disease. Support: ACVO Vision for Animals Foundation. ABSTRACT NO.: 026 The analgesic effect of intracameral lidocaine on intraoperative and postoperative pain for phacoemulsification in dogs S. A. Park,* Y. W. Park,* W. G. Son,† T. H. Kim,* J. S. Ahn,* J. T. Ahn,* S. E. Kim,* I. Lee† and K. M. Seo* *Department of Veterinary Ophthalmology, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea, †Department of Veterinary Anesthesiology, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea Objective: To evaluate the intra- and post-operative analgesic effect of intracameral lidocaine for phacoemulsification in dogs Methods: Twelve healthy beagle dogs were randomly assigned to one of two treatments: 2% lidocaine 0.3 ml or BSS 0.3 ml. Pre-anesthetic medication included acepromazine, cefazoline, and topical tropicamide. Anesthesia was induced with propofol and maintained with isoflurane. The initial end-tidal isoflurane concentration was maintained at 1.2%. Heart rate, respiratory rate, arterial blood pressure, body temperature, inspired and end-tidal isoflurane concentration, and oxygen saturation were recorded every 5 min. The allocated agent was injected intracamerally following aspiration of the same volume of aqueous. Ten minutes after injection, phacoemulsification was performed. After the start of surgery, the isoflurane concentration was adjusted according to heart rate and mean arterial blood pressure. Pain scores were recorded before surgery and at 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, 8, 16, and 24 h after extubation. Any dog that had a total pain score of ‡9 or that scored ‡3 in any category received tramadol intravenously. Results: Isoflurane requirements were significantly higher in the BSS group compared to the lidocaine group. The mean time to treatment failure was significantly shorter in the BSS group (1.4 ± 1.2 h) compared to the lidocaine group (4.9 ± 1.2 h; P = 0.014). Conclusions: Intracameral lidocaine had a significant intra- and post-operative analgesic effect in dogs undergoing cataract surgery. This study illustrates the value of intracameral lidocaine as an analgesic for intraocular surgery in dogs. Supported by BK21 program. ABSTRACT NO.: 027 Retinoid replacement therapy with 9-cis retinal improves visual performance in RPE65 mutant dogs P. M. Gearhart,* D. A. Thompson,† C. C. Gearhart* and S. M. Petersen-Jones* *Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI, USA,†Department of Ophthalmology and Visual Sciences, Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI, USA Purpose: Briard dogs carrying a naturally occurring 4-bp mutation in the visual cycle gene RPE6 display loss of visual function in dim light and are a valuable large animal model for use in developing therapeutic strategies for this type of inherited retinal disorder. RPE65 acts an isomerohydrolase to convert esters of vitamin A stored in the retinal pigment epithelium (RPE), to 11-cis retinol that is oxidized to 11-cis retinal and transported to the retina where it combines with rhodopsin and cone-opsins. The purpose of our study is to determine efficacy of retinoid therapy in the RPE65 mutant dog. Methods: 7 RPE65-/- mutant dogs underwent intravitreal injection of 9-cis retinal, a stable analog of 11-cis retinal. Full-field flash ERG analysis and vision evaluation (either subjective or objective) were performed one week after injection. 2 dogs underwent ERG analysis at weekly intervals for 4 and 12 weeks respectively. Two dogs had repeat intravitreal 9-cis retinal injection in the same eye, and two had repeat injections in the contralateral eye. Results: ERG analysis showed improved ERG responses in 6 of 7 dogs following a single injection of 9-cis retinal. Repeat injection in the same eye yielded modest improvement in ERG responses for both dogs in which it was performed. Repeat injection in the contralateral eye improved the ERG of 1 of the 2 dogs for which it was performed. Of 2 dogs undergoing subjective vision evaluation following a single 9-cis retinal injection, 1 dog showed subjective improvement following intravitreal 9-cis retinal injection and one dog was inconclusive, although both dogs showed improved ERGs. Of 2 dogs undergoing objective vision 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 394 a b s t r a c t s testing, both dogs showed improved vision following intravitreal 9-cis retinal injection. Repeat injection in the same eye in both of these dogs showed no improvement in vision testing. Conclusions: Intravitreal injection of 9-cis retinal results in improved ERG responses and visual performance in dim-light. Possible causes for differences in degree of ERG response or lack of improvement in some cases may include age of the animal and diffusion of retinoid from the injection site within the vitreous. Supported by Glassen Foundation (SMPJ, MSU CVM Purebred Dog Endowment Fund, Michigan Eye Bank, Foundation Fighting Blindness, Research to Prevent Blindness. ABSTRACT NO.: 028 The effect of intracameral tissue plasminogen activator (TPA) administration on short-term intraocular pressure (IOP) following pseudophakic cataract surgery in canine patients D. W. Esson, M. D. Armour, J. L. Brinkis, M. M. Fife and S. T. Calvarese Eye Care for Animals, Tustin, CA, USA Purpose: Phacoemulsification is recognized as the optimal technique for canine cataract removal. Postoperative ocular hypertension (POH) is a well-recognized surgical complication. It has been proposed that POH may result from an accumulation of inflammatory and fibrinous debris in the aqueous outflow tracts. Intraocular fibrinolysis using TPA has been documented in dogs. The purpose of this study was to evaluate the effect of intracameral TPA administration on short-term postoperative IOP. Methods: A total of 40 eyes of 30 dogs were randomly assigned to receive an injection of either 0.2 ml TPA or 0.2 ml Balanced Salt Solution (BSS) intracamerally, at the conclusion of pseudophakic cataract surgery. All surgeries were performed by the same DACVO. Patients underwent complete ophthalmic examinations prior to surgery. IOP, anterior chamber (AC) flare and AC cell were recorded on day 0 (6 h post-operatively), day 1 and day 7. Results: TPA; day 0 (average IOP = 14.35 mmHg, flare = 2.35 & cell = 2.05), day 1 (average IOP = 13.4 mmHg, flare = 2.8, cell = 2.5) & day 7(average IOP = 12.6 mmHg, flare = 0.5, cell = 0.35). BSS; day 0 (average IOP = 13.85 mmHg, flare = 2.23 & cell = 1.85), day 1 (average IOP = 15.45 mmHg, flare = 2.4, cell = 1.95) & day 7(average IOP = 12.2 mmHg, flare = 0.6, cell = 0.75). The incidence of POH (>25 mmHg) was 10% (2/20) in both groups. No significant differences between groups were noted when IOP, AC flare and cell were compared, using both single factor ANOVA and a two sample t-test. Conclusions: Intracameral TPA administration during pseudophakic cataract extraction had no effect on the incidence of POH and no statistically significant effect on short-term post-operative IOP, cell or flare. Funding: N/A ABSTRACT NO.: 029 Anterior segment pathology and corneal wound healing in Pax6 heterozygote mice P. J. Accola,* P. A. Moore,* K. P. Carmichael† and J. D. Lauderdale‡ *Department of Small Animal Medicine and Surgery, University of Georgia, College of Veterinary Medicine, Athens, GA, USA, †Department of Pathology, University of Georgia, College of Veterinary Medicine, Athens, GA, USA, ‡Cellular Biology, University of Georgia, Athens, GA, USA Purpose: Establish corneal wounding model and characterize corneal wound healing and anterior segment pathology in Small eye (Sey) (Pax6+/-) mice. Methods: The anterior segment of Wild Type (WT) (n = 84) and Sey (n = 40) mice (age 2–3 months) were examined with slit lamp biomicroscopy. Corneal wounding with n-heptanol was performed and healing evaluated on days 1, 2, 3 (if fluorescein positive on day 2), 4, 7, 14, and 28 post-wounding. Each day, 3–5 mice were euthanized. Anterior segment histopathology was characterized. Immunohistochemistry (p63, sVEGFR-1) was performed. Days to negative fluorescein staining were compared using the Chi-square test. The number of p63 staining basal cells was recorded and statistically evaluated by ANOVA. Staining for sVEGFR1 was recorded as positive or negative Results: Ophthalmic examination and histopathologic findings in Sey mice included corneal opacity, corneal vascularization, iris hypoplasia, and cataract. The wounding protocol resulted in removal of corneal epithelium. There was no significant difference in the amount of cornea wounded in WT and Sey mice (P > 0.05). There was a statistically significant delay in corneal wound healing in Sey mice at days 2 and 3 when compared to WT mice (P < 0.05). There was no significant difference in p63 staining (P > 0.05). All mice exhibited comparable sVEGFR1 staining. Conclusions: Histopathologic findings are consistent with impaired anterior segment development. The delayed corneal healing is unlikely to be due to a deficiency in p63 cellular expression. The comparable expression of sVEGFR1 suggests that it alone is likely not responsible for the corneal vascularization present in Sey mice. Supported by University of Georgia, College of Veterinary Medicine, Veterinary Ophthalmology Research Funds; Children’s Glaucoma Foundation. ABSTRACT NO.: 031 Effect of three treatment protocols on acute ocular hypertension after phacoemulsification and aspiration of cataracts in dogs M. Crasta,* A. B. Clode,† R. J. McMullen,† D. O. Pate† and B. C. Gilger† *Animal Eye Clinic Visionvet, Bologna, Italy, †Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA Purpose: To compare the effect of topical latanoprost, intracameral carbachol, or no adjunctive medical therapy on the development of acute postoperative hypertension (POH) and inflammation after routine phacoemulsification and aspiration (PA) of cataracts in dogs. Methods: A total of 82 dogs (159 eyes) received either one drop of topical 0.005% latanoprost, an intracameral injection of 0.3 ml of 0.01% carbachol, or no adjunctive therapy immediately following PA of cataract(s). Intraocular pressure (IOP) was measured in all dogs 2 and 4 h after surgery. IOP was measured and aqueous flare assessed at 8 AM the day after surgery. Results: Carbachol-treated dogs had significantly higher mean IOP (33.2 ± SD 20.8 mmHg) 2 h after surgery than the dogs receiving no adjunctive therapy (22.0 ± SD 14.1 mmHg) (P = 0.049). There were no significant differences in the number of POH episodes between dogs treated with carbachol (47%), latanoprost (29%), or dogs that received no adjunctive therapy (33%). There were no significant differences in mean aqueous flare grade between eyes treated with latanoprost (1.7 ± SD 0.4) or carbachol (1.4 ± SD 0.6), and eyes that received no adjunctive therapy (1.7 ± SD 0.4). Conclusions: Topical 0.005% latanoprost or intracameral injection of 0.3 ml of 0.01% carbachol after PA in dogs did not reduce POH or cause increased intraocular inflammation compared to dogs not receiving adjunctive therapy after PA of cataracts. ABSTRACT NO.: 033 A procedure for the correction of eyelid agenesis N. M. Bromberg* and K. Caruso† *SouthPaws Ophthalmology, Fairfax, VA, USA †VCA/Veterinary Referral Associates, Rockville, MD, USA Purpose: Various procedures have been described to treat eyelid agenesis, including cryoepilation of trichiasis, lip to lid transposition, pedicle flap and cross-lid flap. We present a procedure to establish a functional upper eyelid. Method: Subjects were shelter cats, with lid defects 50–90% of the lateral upper eyelid. The surgical procedure is performed in two stages. The recipient site is prepared, leaving as much conjunctiva as possible. A flap the length of the upper eyelid defect is made by full-thickness incision of the lower lid from the lateral canthus medially. The lateral aspect of the lower eyelid flap is rotated 180 and sutured into the upper eyelid recipient area aligning the newly created lid margin using a figure-eight 6–0 non-absorbable suture. Conjunctiva is sutured with absorbable suture. Then non-absorbable sutures are used to suture the flap to the recipient site, leaving a keyhole for vision. In the second stage an incision extending beyond the lower eyelid margin separates the new upper eyelid from the lower eyelid. Palpebral conjunctiva is sewn over the cut edge of the lower eyelid. Upper eyelid canthus formation requires cutting a small triangle from the dorsolateral aspect of the new upper eyelid and suturing it in place. Results: This procedure resulted in cosmetic, functional upper eyelids with normal conjunctival lining. Corrections for trichiasis or to close small gaps, usually at the recipient-graft connection, were occasionally necessary. Two cases required cicatricial entropion correction of the upper eyelid. The lateral aspect of the lower eyelid healed with a smooth, hairless, non-irritating edge. Discussion: A procedure for eyelid agenesis correction is presented. Benefits include a functional upper eyelid with normal conjunctival surface, smooth lateral lower eyelid margin, and hairs in the proper alignment. Complications include an irregular upper eyelid margin and/or trichiasis at the connection point, or cicatricial entropion. ABSTRACT NO.: 034 Early and late treatment of corneal neovascularization by subjonjunctival injection of bevacizumap in a rabbit model of limbal stem cell deficiency K. T. Kuo,* W. L. Chen,† N. T. Lin,* F. R. Hu† and C. T. Lin* *School of Veterinary Medicine, National Taiwan University, Taipei, Taiwan, †Department of Ophthalmology, College of Medicine, National Taiwan University, Taipei, Taiwan Purpose: To compare the difference of early and late subconjunctival injection of bevacizumab in the treatment of corneal neovasularization (NV) and conjunctivalization in a rabbit model of limbal insufficiency. Methods: After creating limbal insufficiency by mechanical limbal injury in rabbits, 2.5 mg subconjunctival bevacizumab was injected twice per week immediately (early treatment group) or 1 month later (late treatment group) for one month. Digital photography was recorded to evaluate the corneal NV and opacity. Immunohistochemical staining of K3,K4, K12, K13 and MUC5 in corneal surface cells was performed at 1 month after treatment. Results: Subconjunctival bevacizumab significantly inhibited the formation of corneal NV in the early treatment group but not in the late treatment group compared to the untreated group. The immunohistochemical study demonstrated that corneal surface cells was K3 (+), K4 (-), K12 (+), K13 (-) and MUC5 (-) in the early treatment group, and K3 (+), K4 (+), K12 (-), K13 (+) and MUC5 (+) in the late treatment group. Conclusions: Early treatment of limbal injury by subconjunctival bevacizumab is more effective than late treatment in inhibiting corneal NV and conjunctivalization manifested by cellular markers of the cornea. The first two authors contributed equally. ABSTRACT NO.: 036 Comparative ocular anatomy of the duck-billed platypus C. J. Zeiss,* I. Schwab,† C. Murphy‡ and R. W. Dubielzig§ *Yale University School of Medicine, New Haven, CT, USA, †University of California School of Medicine, San Francisco, CA, USA, ‡University of California School of Veterinary Medicine, Davis, CA, USA, §University of Wisconsin School of Veterinary Medicine, Madison, WI, USA Purpose: To identify evolutionary origin and fate of anatomic features of the duck-billed platypus eye. Methods: Eyes from a single platypus and a broad range of other vertebrates (including hagfish, lamprey and lungfish) were prepared for light microscopy. In addition to a standard panel of special stains, tissues were immunostained against a variety of rod and cone opsins. Finally, published opsin sequences of platypus and other key species were aligned and compared with immunohistochemical results. Results: A complete scleral cartilage similar to that seen in birds, reptiles and amphibians encloses the platypus eye. This feature is present in sharks and rays, and in the earliest extant tetrapod, the lungfish. The ciliary body is large and devoid of a pars plana. Zonular ligaments are invisible by light microscopy, suggesting an eye of minimal accommodative capacity. The choroid lacks a tapetum. The retina is largely avascular and is rod-dominated, with a minority of red- and blue-cone immunoreactive photoreceptors. Like marsupials and many non-mammalian species, cones contain clear inner segment droplets. Double cones were present, a feature not found in eutherian mammals or marsupials. Ganglion cell density is equivalent to that of a mouse. Evaluation of opsins indicates that red and blue immunoreactive cone opsins, but not rhodopsin, are present in the most primitive extant species examined – the hagfish. Rhodopsin appears in the earliest extant tetrapod examined, the lungfish. Conclusions: Unlike eyes of eutherian mammals, the platypus eye has retained morphologic features present in very early tetrapods such as the lungfish and amphibians. These include scleral cartilage, double cones and cone droplets. In the platypus, as in other mammals, rhodopsin is the predominant photoreceptor pigment, at expense of the cone system. 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 abstracts 395 ABSTRACT NO.: 037 Effect of an embroyonic like bioscaffold in dogs with persistent indolent ulcers J. Urbanz* and C. W. Woods† *Eye Care for Animals, Scottsdale, AZ, USA †TR BioSurgical, LLC, Scottsdale, AZ, USA Purpose: This study’s purpose is to evaluate a novel bioscaffold’s corneal repair properties in dogs with persistent indolent ulcers. The bioscaffold consists of a unique collagen co-polymer that structurally resembles the tertiary structure of the embryonic extracellular matrix; thereby exposing a natural set of amino acid sequences that cause infiltrating cells to up-regulate ‘‘fetallike’’ repair mechanisms. Methods: This is an ongoing prospective, controlled field study in 20 dogs with persistent indolent ulcers, randomly assigned into bioscaffold and control groups. After corneal debridement, all patients receive standard ophthalmic medications, with one group receiving the bioscaffold preparation as a topical powder once daily for three consecutive days. Outcomes for assessment consist of surface area of lesion and degree of keratitis, conjunctival hyperemia, and blepharospasm on days 1 and 14 using a standardized numerical score system. Results (interim): A total of 15 patients have completed the study, 6 bioscaffold patients and 9 control patients. At day 14, only 33% of control dogs had complete corneal healing compared to 67% from the bioscaffold group. The bioscaffold group had a 67% reduction in blepharospasm compared to only 18% in the control. All other outcome measures were equivalent at this stage. Conclusions: These interim data support the embryonic-like bioscaffold improves corneal healing and reduces blepharospam. One possible explanation is the bioscaffold provides a structural matrix for infiltrating cells such stem cells, progenitor cells and fibroblasts to hasten corneal surface repair. Funding Sources: TR BioSurgical, LLC P, E ABSTRACT NO.: 039 Effect of tapetum on vision in extremely low light in labrador retrievers M. H. Neaderland* and L. M. Occelli† *Animal Eye Clinic, Norwalk, CT, USA, †Alluch, France Purpose: To determine if the presence or absence of an ophthalmoscopically visible tapetum affects the performance of a Labrador Guide Dog in extremely low light conditions. Methods: Digital retinal photography, indirect and slit lamp ophthalmoscopy were performed on the eyes of 214 Labrador Retrievers, the breeding stock for Guiding Eyes for the Blind. The appearance of each tapetum was classified as T-full tapetum, MT-medium tapetum or A- no visible tapetum. Ten dogs from each category of tapetum were selected with a mix of sexes and coat color for each group. Two timed tests were performed in the light and dark on each dog. Test 1-maze test-a corridor of randomly varied obstacles, Test 2-tunnel test – a box with 3 potential outlets which were randomly opened for dog to exit through. Results: All dogs took longer to perform the tests in dim light. No significant difference was found between the times it took for the three categories of tapetum to navigate either test in light nor dark. Conclusion: The visible variability of the tapetum of the Labrador Retriever has no effect on the ability of the dog to perform in extremely low light. ABSTRACT NO.: 040 Superficial keratectomy for the treatment of equine immune mediated keratitis D. O. Pate, A. B. Clode, R. M. McMullen, J. H. Salmon and B. C. Gilger North Carolina State University, Raleigh, NC, USA Purpose: Immune-mediatedkeratitis(IMMK)isacommon,chronic,relapsing inflammatorydisease of the cornea in horses. It is likely driven by an autoantigen located at the site of inflammation, althoughtheexactetiologyisunknown.Thepurposeofthisstudywastodeterminetheeffectiveness ofremovalofthelesion(andpossiblytheautoantigen)viasuperficialkeratectomy(SK)asatreatment for superficial and midstromal equine IMMK. Methods: Horses that were diagnosed with chronic superficial or midstromal IMMK, based on criteria described in Gilger BC, et al., Vet Ophthalmol 2005, had a standing superficial keratectomy. Keratectomy tissues were collected, fixed, and processed for histopathology. Horses were followed after surgery for complications and evidence of recurrence of IMMK. Results: Data from 13 eyes of 12 horses diagnosed with IMMK that had a superficial keratectomy was reviewed. There were no recurrences of IMMK after superficial keratectomy. Histopathology revealed mononuclear cellular infiltrate but no evidence of microorganisms. Conclusions: For superficial or midstromal IMMK in horses, a superficial keratectomy may be beneficial for the long-term control of the disease. It is likely that this therapy is helpful because of the surgical removal of the autoantigen (s). Further work is needed to determine the types of autoantigens present in these eyes and the immunologic mechanisms responsible for the chronic keratitis. Support: State of North Carolina, North Carolina Uveitis Research Fund. Commercial Relationship: Pate DA (None), Clode AB (None), McMullen RM (None), Salmon JH (None), Gilger BC (None) ABSTRACT NO.: 041 Retinal/cortical imaging in dog retinal degeneration models G. D. Aguirre,* G. M. Acland,† G. K. Aguirre,‡ T. S. Alemán,§ W. A. Beltran,* A. V. Cideciyan,§ W. W. Hauswirth,– S. G. Jacobson§ and A. M. Komáromy* *Ophthalmology, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA, USA, †Baker Institute, Cornell University, Ithaca, NY, USA, ‡Neurology and §Ophthalmology, School of Medicine, University of Pennsylvania, Philadelphia, PA, USA, –Ophthalmology, University of Florida,Gainesville, FL, USA Purpose: Many primary diseases of the photoreceptor or RPE in the dog are caused by mutations in genes that cause inherited diseases in human patients, and the phenotypes in both species are strikingly similar. Coupled with recent advances in gene therapy that show rescued function and preservation of retinal structure, the dog has become an important species to investigate disease mechanisms, and facilitate translation of new therapies to human patients. To facilitate these studies, we describe the use of different imaging modalities in dogs that are used in the assessment of patients with inherited retinal diseases. Methods: We have used a combination of functional magnetic resonance imaging (fMRI), optical coherence tomography (OCT) and infrared/autofluorescence imaging of the retina in dogs having naturally occurring mutations in the RPE65, RPGR, RHO and CNGB3. Mutations in these genes, respectively, cause LCA, XLRP, AdRP and achromatopsia. In the RPE65 and CNGB3 models, assessment has been done before and after successful gene therapy. These results have been correlated with ERG and morphologic evaluation. Results: There is excellent correlation between the different imaging modalities and the functional/structural outcomes in the naturally occurring diseases, and following successful retinal gene therapy. Comparison of in vivo with the ex vivo results provide micron level resolution between histology and OCT. Conclusions: The different imaging approaches show excellent correlation with the histopathology, and provides a basis for improved interpretation of the non-invasive measurements. The ability to assess both the human patient and the animal model with the same in vivo imaging modalities will allow for a better understanding of the disease processes, and facilitate future translational studies. Supported by: NEI/NIH grants EY-06855, -17549, -19304, FFB, The ONCE International Prize, and the Van Sloun Fund. ABSTRACT NO.: 042 Novel VMD2 mutation(s) in canine multifocal retinopathy G. D. Aguirre,* S. J. Lindauer,* K. Wickström,† S. Ahonen,‡ H. Lohi,‡ A. Gupta,* G. M. Acland,§ K. Guziewicz* and B. Zangerl* *Ophthalmology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA, †Veterinary Clinic Akuutti, Finland, ‡Department of Basic Veterinary Sciences and Department of Medical Genetics, Folkhälsan Institute of Genetics, University of Helsinki, Helsinki, Finland, §Baker Institute, Cornell University, Ithaca, NY, USA Purpose: Mutations in the VMD2 gene are associated with various retinopathies in man and dog; in the latter species the disease termed canine multifocal retinopathy (cmr). The ability to pursue detailed molecular and clinical studies in the dog to investigate disease mechanisms and potential therapeutic approaches for disorders related to mutations in the VMD2 gene, make the dog an important model organism to carry out these studies. To investigate the influence of bestrophin related retinopathies in the dog, and identify additional disease variants, selected dogs with a retinal phenotype compatible with cmr were screened for mutations in VMD2, and identified changes evaluated for disease association. Methods: DNA was isolated following standard protocols from blood or buccal swabs of Lapponian Herder dogs (LHs) from Finland with ascertained clinical diagnosis. Coding exons of VMD2 were amplified as previously described (Guziewicz et al., 2006), and sequences were aligned to published wild type sequence. Results: LHs originating in Finland display a disease, formerly termed retinopathy, that is clinically identical to previously described canine multifocal retinopathy (cmr1, cmr2). Unlike other breeds with known cmr mutations, affected LHs carry two homozygous mutations in exon 10; a deletion leading to a frame shift (CFA18, bp 57499959, Cdel, Pro463FS), and a point mutation (CFA18, bp 57499881, G > T). The latter leads to a stop codon at aa 490 if coupled with the deletion, or a Gly489Val substitution in the absence of the deletion. Within the investigated pedigree, these two mutations are completely linked, and associated with the disease phenotype. Neither of the mutations was found in a screen of samples from dogs with retinal changes of 25 additional breeds from Finland. Conclusions: We describe an additional mutation in the VMD2 gene in the Lapponian Herder breed, and refer to the involved disease locus as cmr3. These results indicate that cmr has a larger impact on the general dog population then was initially thought, and will further contribute to ongoing studies to understand the molecular basis of mutations in VMD2 and define correlations of observed genotypes with the respective phenotypes. Support: FFB, NEI/NIH EY06855, MAF, Van Sloun Fund, Academy of Finland, Sigrid Juselius Foundation. P, I (Aguirre, Acland). ABSTRACT NO.: 043 The effect of intravitreal ciliary neurotrophic factor (CNTF) on structure, function and gene expression of normal canine retinas A. T. Parton-Corr, G. D. Aguirre, W. A. Beltran and A. M. Komáromy Small Animal Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA Purpose: Determine the short-term effect of intravitreal CNTF on macroscopic and microscopic morphologic retinal changes, global retinal function, and expression and localization of phototransduction cascade components in rods and cones of the normal canine retina. Methods: After baseline ophthalmic examination and electroretinogram, eight normal canine eyes received an intravitreal injection of 12 lg (30 lg) CNTF. The contralateral eyes were injected with PBS and served as controls. Final ocular examination, electroretinography and tissue collection occurred at 1, 2, and 5 weeks. Retinal cryosections were evaluated morphologically with H&E stain. Fluorescence immunohistochemistry was used to evaluate human cone arrestin (hCAR), short- (S) and long/medium (L/M) wavelength cone opsins, rhodopsin and cone alpha transducin. Quantitative real-time polymerase chain reaction was performed to assess expression of S opsin, L/M opsin, rhodopsin, and rod and cone cyclic nucleotide gated channel subunits (CNGA1, CNGB1, CNGA3, CNGB3). Results: Intravitreal CNTF caused shortening and disorganization of photoreceptor outer segments evident by 1 week post-injection with recovery evident by at least 5 weeks post-injection. Compared to control eyes, ERGs of CNTF injected eyes showed severely decreased function 1 week post-injection with recovery evident by 2 weeks post-injection and function comparable to baseline by 5 weeks post-injection. In CNTF injected eyes there was a down-regulation of gene expression of rhodopsin, S-opsin, L/M opsin, CNGA1, CNGB1, CNGA3 and CNGB3 to 18.5%, 0.1%, 32%, 18.8%, 19.3%, 24.6% and 16.6% of controls, respectively, evident by 1 week post-injection with recovering expression by 5 weeks post-injection. These results are consistent with those seen previously in rodent models. Conclusions: Intravitreal CNTF causes a transient photoreceptor deconstruction within 1 week and photoreceptor reconstruction beginning by at least 2 weeks post-injection. Photoreceptors return to near normal/normal by 5 weeks post-injection. Photoreceptor reconstruction after the transient photoreceptor deconstruction may provide an opportunity for increasing the success of gene therapy in the treatment of inherited retinopathies. Supported by the ACVO Vision for Animals Foundation, NIH grants EY006855, EY017549, EY019304, K12EY015398 and P30EY001583, Foundation for Fighting Blindness. 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 396 a b s t r a c t s ABSTRACT NO.: 044 Aqueous humor vascular endothelial growth factor in dogs with intraocular disease C. A. Sandberg,* I. P. Herring,* W. R. Huckle,† T. LeRoith,† J. Phillip Pickett* and J. H. Rossmeisl* *Department of Small Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Blacksburg, VA, USA, †Department of Biomedical Sciences and Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Blacksburg, VA, USA Purpose: To examine levels of vascular endothelial growth factor (VEGF) in aqueous humor (AH) of dogs with intraocular disease and to evaluate the association of AH VEGF levels with pre-iridal fibrovascular membrane (PIFM) formation. Methods: Aqueous humor samples (n = 226 from 178 dogs) and matched plasma samples (n = 101) were collected from dogs with a variety of intraocular diseases (including primary cataract, diabetic cataract, primary glaucoma, uveitic glaucoma, aphakic/pseudophakic glaucoma, retinal detachment, lens luxation and intraocular neoplasia), as well as 13 ophthalmically normal control dogs. Systemic disease status, corticosteroid administration and ophthalmic prostaglandin administration were recorded for clinical cases. Aqueous and plasma VEGF was assayed via ELISA validated for canine VEGF. Available histopathology samples were examined for the presence of PIFM and categorized as none, cellular, fibrous or vascular, and fibrovascular. Data were evaluated by mixed model ANOVA, with application of Tukey-Kramer adjustment for multiple comparisons. Results: Compared to normal controls, AH VEGF was significantly elevated for all intraocular diseases (P < 0.05) except for primary and diabetic cataracts. Systemic disease had no significant effect on AH VEGF (P < 0.05), but corticosteroid and prostaglandin administration significantly affected AH VEGF (P < 0.05). Compared to dogs without PIFM, AH VEGF was significantly higher in dogs with fibrovascular PIFM (P = 0.001), but not cellular (P = 0.1704) or fibrous/vascular PIFM (P = 0.0667). There was no association between AH and plasma VEGF levels. Conclusions: These results are consistent with the hypothesis that AH VEGF plays a role in the causation or progression of a variety of canine intraocular disorders, including the development of PIFM. ABSTRACT NO.: 045 Evaluation of the ocular penetration of topical GVT E. G. da Silva,* J. R. Gionfriddo,* S. F. Hudacheck,* D. L. Gustafson,* F. Olea-Popelka,* C. C. Powell* and V. L. Scofield† *Department of Clinical Sciences, Colorado State University Veterinary Medical Center, Fort Collins, CO, USA, †Department of Carcinogenesis, The University of Texas M.D. Anderson Cancer Center, Science Park-Research Division, Smithville, TX, USA Purpose: Oxidative stress plays a major role in the pathogenesis of many neuroimmunodegenerative diseases. The accumulation of reactive oxygen specieshasalso been implicated as one of the pathogenic mechanisms causing retinal and optic nerve changes in glaucoma. GVT has shown profound anti-inflammatory and antioxidant effects, both in experimental animals and human clinical studies. To our knowledge, GVT has never been used topically. The aim of this study was to investigate the ocular penetration of topical GVT. Methods: Eleven rabbits were divided into three groups of three treated animals each, and one group of two untreated animals. Each treatment group received one drop of GVT of different concentrations in both eyes (0.5%, 1.5%, 2.5%). Aqueous humor and peripheral blood samples were taken at three different timepoints (20 min, 4 h and 12 h). At the last timepoint, enucleation was performed; lens, vitreous and retinal sampleswere harvested. Samples were analyzed using High Performance Liquid Chromatography (HPLC). Results: GVT was detected in the aqueous humor of all treated rabbits and at all timepoints. It was found in the vitreous and in the peripheral blood in the 1.5% and 2.5% concentration groups, at 12 h. The highest aqueous humor concentration of GVT was 231 ng/ml after 20 min, 47.9 ng/ml after 4 h and 13.2 ng/ml after 12 h. Conclusions: Topically administered GVT readily penetrates the anterior chamber and vitreous chamber as well, making it potentially available for prevention of oxidative stress in retinal cells. This study was supported by a grant from the Longevity Foundation. GVT was kindly provided by Bach Pharma. ABSTRACT NO.: 046 Recurrent superficial keratitis in a male white Asian elephant living in a closed stable A. Thayananuphat,* P. Diskul,† S. Mahasawangkul‡ and P. Santitiseri* *Faculty of Veterinary Medicine, Kasetsart University, Thailand, †Royal White Elephant’s Stable, Royal Chitrlada Palace, Thailand, ‡Thai Elephant Conservation Center, National Elephant Institute, Thailand Purpose: Superficial keratitis is rarely described in Asian pachyderm (elephant). Due to the size of this animal, ophthalmic approach to such problem could be troublesome. We describe herein a case of superficial keratitis in a male white elephant (Elephas maximus) living in a closed stable. Methods: A 58 year old male white Asian elephant presented with blepharospasm of the right eye and epiphora of both eyes. Routine ophthalmic examination was performed. Ophthalmic lesions were investigated with difficulty, therefore we decided to determine his lesions via photographs captured by a digital camera (Nikon, D70S, Thailand) with a high magnified power lens (Nikon, AF Micro-Nikkor 105mm f/2.8D, Japan). Results: High resolution pictures have indicated superficial keratitis of both eyes with a shallow corneal ulcer of the right eye. Corneal swab culture further revealed the growth of Staphylococcus spp. Due to his special treatment as a white elephant, he lived in a closed-space stable where we have smelt strong presence of ammonia, possibly from his urination. It is quite likely that this might be a cause of his irritated keratitis. He received systemic treatment with intramuscular injection of phenylbutazone and topical medication with triple antibiotic solution. Topical corticosteroid was provided only when the ulcer was healed. We also recommended his caretakers to remove him into an open-space stable with better air ventilation during the day and increase sanitation within the stable. Miraculously, his eye problems were rapidly resolved. However, recurrent superficial keratitis was observed within 4 months later. For the second episode, he received topical corticosteroid and artificial tears and has completely moved into an open-space stable. After successful treatment of his second episode, no recurrence of superficial keratitis was observed for one year follow up period while the elephant completely remained in the new environment. Conclusions: To the best of our knowledge, this is the first report of superficial keratitis in a white elephant. Our approach for ophthalmic examination by high resolution photography used in this elephant could be applicable for future cases. Owing to the fact that the special treatment he received might play an important role on developing this rare condition, a combination of medical therapy and environmental control are both required to provide the best outcome possible. ABSTRACT NO.: 047 Topical nalbuphine or oral tramadol as analgesics for corneal pain in dogs: a pilot study J. S. Clark,* E. Bentley† and L. J. Smith† *Eye Care for Animals, IL, USA, †Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI, USA Purpose: To compare the effectiveness of oral tramadol and topical nalbuphine in the treatment of corneal pain in dogs. Methods: A total of 14 male dogs were divided into three treatment groups. Dogs were sedated with dexmedetomidine (5 lg/kg IV) and a 4 mm corneal epithelial wound was created in the right eye(OD) of all dogs. Sedation was reversed with atipamazole IM. All dogs received pre/post ophthalmic examinations. Post operatively, Group NB (n = 5) received topical 1% preservative-free nalbuphine OD q8 hours and a placebo tablet PO q8 hours. Group TR (n = 5) received tramadol (4 mg/kg) PO q8 hours and topical sterile saline OD q8 hours. Group CNTRL (n = 4) received topical sterile saline OD q8 hours and an oral placebo tablet q8 hours. Dogs received gentamicin (0.3%) OD BID. Dogs were pain scored at 0,1, 2, 4, and 6 h then q6 hours by observers masked to treatment until corneal wounds were healed. Treatment failure was recorded if cumulative pain scores were ‡9 or a category score was ‡3 and rescue analgesia (morphine 1.0 mg/kg IM) was administered. The incidence of treatment failure was analyzed using Fisher’s exact test Results: Rescue analgesia was required by 4 dogs in Group NB, 1 dog in Group TR, and 2 dogs in Group CNTRL. There was no significant difference between groups (P = 0.184). Mean time to rescue was 9.16 h. All corneal wounds were healed by 84 h. Conclusions: Although not statistically significant, tramadol appears to be a viable option for the treatment of corneal pain. Topical nalbuphine appears to be less effective than saline when used for treatment of corneal pain, as demonstrated by the high incidence of treatment failure in dogs receiving nalbuphine. ABSTRACT NO.: 048 Stability of 0.5% cidofovir under various storage conditions for up to 6 months J. Stiles,* R. M. Pogranichniy† and W. E. Gwin‡ *School of Veterinary Medicine, Purdue University, West Lafayette, IN, USA †Department of Veterinary Clinical Sciences, Veterinary Teaching Hospital Pharmacy, West Lafayette, IN, USA, ‡Department of Comparative Pathobiology, Veterinary Teaching Hospital Pharmacy, West Lafayette, IN, USA Purpose: To evaluate the effect against feline herpesvirus (FHV-1) of 0.5% cidofovir solution when stored for up to 6 months. Methods: Cidofovir (Vistide Gilead USA) was diluted with normal saline to a 0.5% concentration. Aliquots were placed in glass and plastic vials and stored at 4C, -20C and -80C. At time zero, the effect of fresh cidofovir was evaluated against FHV-1 in a virus titration assay. At time points 30 d, 60 d, 120 d and 180 d, cidofovir under each of the different storage conditions was used in a virus titration assay to assess stability of the drug. Results: At each time point and under all storage conditions cidofovir reduced viral titer by 4 logs (P < 0.001). There was no effect of time, storage temperature, or storage vial material on the anti-viral activity of the drug (P > 0.05). Conclusions: Cidofovir 0.5% solution is stable for up to 6 months in either glass or plastic at 4C, -20C and -80C. ABSTRACT NO.: 049 A histologic series of secondary uveal melanomas in eleven dogs K. A. Konrade,* A. Hoffman,* C. S. Schubert† and R. R. Dubielzig† *Eye Care For Animals, Pasadena, CA, USA, †School of Veterinary Medicine, University of Wisconsin, Madison, WI, USA Purpose: To describe the clinical and histological features of a series of eleven canine cases of secondary uveal melanomas. Methods: Eleven canine cases of secondary uveal melanomas were histologically examined at the comparative ocular pathology laboratory of Wisconsin (COPLOW). Age, primary site of origin, time to metastasis, presenting ocular clinical signs, and histological descriptions were examined. Results: Dogs were between 6 and 15 years, with an average age of 9 years. Most were large breed dogs and distant primary tumor sites were of the oral cavity, distal limbs (digits), and skin. Prior to enucleation, 10/11 cases showed signs of anterior uveitis and 4/11 were diagnosed with secondary glaucoma. 4/11 cases presented with anterior uveal masses of varying pigment (light tan to dark brown). 8/11 tumors presented in the iris and/or ciliary body and 3/11 presented in the iris/ciliary body and choroid. All of the eyes with secondary choroidal tumors had evidence of retinal detachment. 6/11 primary tumors arose from the digits, 3/11 were from the oral cavity (palate or gingival) and 2/11 were cutaneous (including the lip). The average time for dogs to develop clinically apparent metastatic disease to the uveal tract was 2 years (range 1 month–4 years) after the initial diagnosis of malignant melanoma. Conclusion: Ocular examination of localized and generalized malignant melanoma in dogs may reveal a far greater number of intraocular metastatic melanomas than previously documented. Furthermore, all dogs with a suspected uveal tumor should receive a thorough physical examination including the digits, oral cavity, and integument even if the uveal mass is in a normotensive, visual eye. ABSTRACT NO.: 050 Combined diode endoscopic cyclophotocoagulation and Ex-Press shunt gonioimplantation in four cases of canine glaucoma E. A. Lutz and J. S. Sapienza Long Island Veterinary Specialists, Plainview, NY, USA Purpose: To evaluate the implantation of an Ex-Press LR-50 shunt in combination with diode endoscopic cyclophotocoagulation (ECP) as a means for blunting post-operative intraocular pressure (IOP) spikes. Methods: Four eyes from four dogs with glaucoma were treated 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 abstracts 397 with a limbal approach ECP (E4 Endo Optiks), as well as Ex-Press shunt gonioimplantation. Preoperative and postoperative clinical courses were retrospectively reviewed. Patients received ECP and Ex-Press gonioimplantation for treatment of primary glaucoma or for treatment of secondary glaucoma refractory to medical therapy (intraocular pressure (IOP)>25 mmHg on medication). Results: Four purebred dogs were treated. Three were neutered males, and one was a spayed female. The right eye was affected in three cases, and the left eye in one case. At the time of surgery, two eyes were pseudophakic, and two were phakic. Phakic eyes underwent phacofragmentation at the time of surgery. Average ECP laser energy delivered was 250 mW at a continuous duration. Average extent of ciliary process ablation was 307.5 ± 15 degrees (range 300–330 degrees). All eyes had Ex-press shunt gonioimplants placed at the time of ECP. Average postoperative observation was 12.3 ± 0.5 months (range 12–13 months). Average pre-operative IOP was 46.3 ± 17.8 mmHg. Average post-operative IOP was 9.8 ± 5.7 mmHg at one day, 14.5 ± 10.4 mmHg at one week, 18.8 ± 17.9 mmHg at two weeks, 8 ± 5.7 mmHg at one month, and 17.5 ± 20.1 mmHg at two months. 1/4 dogs experienced an IOP spike at one week post-operatively, and developed glaucoma by two weeks post-operatively until 2 months postoperatively. 3/4 dogs experienced normotensive IOP (range < 5 to 19 mmHg) during the immediate two month post-operative period, with no IOP spikes recorded. IOP remained normotensive (< 25mmHg) after ECP in 2/4 eyes from 24 h postoperatively to last evaluation. Postoperative long-term complications with this combined procedure included recurrence of glaucoma (1/4), and hypotony (1/4). Conclusions: In the treatment of canine glaucoma, the combined procedures of ECP and Ex-Press shunt gonioimplantation were associated with normotensive IOP and no recorded IOP spikes in the immediate two month post-operative period in 3/4 eyes. ABSTRACT NO.: 051 Diode endoscopic cyclophotocoagulation in pseudophakic and aphakic dogs with secondary glaucoma E. A. Lutz and J. S. Sapienza Long Island Veterinary Specialists, Plainview, NY, USA Purpose: To evaluate the use of a diode laser for endoscopic cyclophotocoagulation in pseudophakic and aphakic dogs with secondary glaucoma following primary cataract removal. Methods: A total of 15 dogs (n = 17 eyes) with secondary glaucoma were treated with a limbal approach endoscopic cyclophotocoagulation (ECP). Preoperative and postoperative clinical courses were retrospectively reviewed. Patients received ECP for glaucoma refractory to medical therapy (intraocular pressure (IOP) > 25mmHg on medication). Results: A total of 14 purebred dogs and one mixed breed dog were treated. Eight were spayed females, six were neutered males, and one was an intact male. Thirteen eyes were pseudophakic and four were aphakic. The right eye was affected in six cases, and the left eye in eleven cases. One dog had previously been treated with transscleral diode laser cyclophotocoagulation in the affected eye. Average ECP laser energy delivered was 250 mW (range 200–350 mW) at a continuous duration. Average extent of ciliary process ablation was 287.1 ± 47 degrees (range 185–360 degrees). Two eyes had Ex-press shunt gonioimplants placed at the time of ECP. Average postoperative observation was 10.6 ± 3.9 months (range 4–17 months). IOP remained normotensive (<25 mmHg) after ECP in 10/17 eyes from 24 h postoperatively to last evaluation. 3/17 eyes required topical prostaglandin analogue therapy for IOP control, and 1/17 eyes developed glaucoma that was controlled after a second ECP treatment. At last evaluation, 7/15 eyes maintained vision, 2/15 eyes regained vision, one eye with previously equivocal vision was non-visual, 4/15 eyes remained non-visual, 1/15 eyes lost vision, and 1/15 eyes became blind secondary to complications unrelated to ECP. On average, three preoperative glaucoma medications were prescribed per patient, one was used at six months postoperatively, and none were administered at twelve months postoperatively. Postoperative complications included superficial corneal ulceration (1/17), recurrence of glaucoma (5/17), and phthisis with blindness (1/17). Conclusions: In secondary pseudophakic and aphakic canine glaucoma, ECP was associated with IOP control and marked decrease in prescribed postoperative glaucoma medications at last evaluation in 16/17 and 14/17 eyes, respectively, and with return or maintenance of vision in 9/15 eyes. ABSTRACT NO.: 053 The effects of topical nalbuphine on canine corneal cells in vitro R. A. Spatola,* M. Thangavelu† and H. L. Chandler*,† *The Ohio State University College of Veterinary Medicine, Columbus, OH, USA, †The Ohio State University College of Optometry, Columbus, OH, USA Purpose: To determine the effects of a 1% topical solution of nalbuphine on canine corneal stromal and epithelial cell viability in vitro. Methods: Primary tissue was obtained from dogs with normal eyes in good general heath, euthanized for population control purposes. Dissection occurred within 1 h post-mortem. Stromal and epithelial cells were cultured separately. Established cells were treated with either a 1% nalbuphine solution prepared in DMEM culture media with 2 mg/mL sodium chloride, or in a vehicle solution contain DMEM and 2 mg/mL sodium chloride. Morphologic characteristics and cellular viability were evaluated over time with microscopy and trypan blue. Results: A 1% solution of nalbuphine was acutely toxic to corneal stromal cells, with marked decrease in cell viability occurring in less than 5 min of administration. This was demonstrated by retraction of cytoplasmic processes, vacuolization and loss of adherence to the culture plate. Canine corneal epithelial cells demonstrated decreased toxicity to treatment with nalbuphine based on morphologic characteristics, with slight vacuolization but maintenance of architecture and monolayer confluence. The drug also proved to be unstable in regards to pH and temperature, with marked precipitation out of solution as temperature and pH approached physiologic levels. Conclusions: A 1% topical ophthalmic solution of nalbuphine has the potential to be toxic to the ocular surface, particularly with regard to eyes containing epithelial defects. Further research is needed to determine if nalbuphine alters corneal wound healing, the metabolic activity of corneal cells, as well as solubility characteristics and drug delivery systems. Supported by Rx Center/Gary Newton, FACA. ABSTRACT NO.: 054 Tear, cornea, and aqueous humor concentrations of ciprofloxacin and moxifloxacin after topical application in normal horses H. D. Westermeyer, D. V. H. Hendrix, D. A. Ward and S. R. Cox College of Veterinary Medicine, University of Tennessee, Knoxville, TN, USA Purpose: Determine ocular tissue drug concentrations after topical administration of 0.3% ciprofloxacin (Falcon Pharmaceuticals, Ltd. Fort Worth, Texas) and 0.5% moxifloxacin (Alcon Laboratories, Inc. Fort Worth, Texas). Methods: Both antimicrobials (0.1 ml) were delivered to the ventral conjunctival fornix of normal horses as follows. Group 1 (n = 8) hours 0, 2, 4, and 6; Group 2 (n = 8) hours 0, 2, 4, 6, and 10; and Group 3 (n = 8) hours 0, 2, 4, 6, 10, and 14. Tears, cornea, and aqueous humor (AH) were collected at 10, 14, and 18 h for Groups 1, 2, and 3, respectively. Drug concentrations were determined by HPLC. Results: The concentration (lg/ml) of ciprofloxacin (median (25%–75%)) for Groups 1, 2, and 3 in tears was 53.67 (25.47–88.75), 48.52 (19.65–74.66), and 24.35 (15.43–67.05), respectively; in cornea (lg/g), 0.945 (0.599–1.02), 0.367 (0.323–0.474), 0.480 (0.340–0.952), respectively; in AH, they were below detection levels in all groups. Concentrations of moxifloxacin for Groups 1, 2, and 3 in tears were 188 (44.5–669.2), 107 (41.7–296), and 178 (70.1–400), respectively; in cornea, 1.841 (1.440–2.109), 0.777 (0.552–0.980), and 0.769 (0.647–0.966), respectively; in AH (lg/ml), 0.0550 (0.037–0.075), 0.0340 (0.0208–0.0475), and 0.0195 (0.006–0.043), respectively. Group 1 corneal moxifloxacin concentrations were significantly higher than Groups 2 and 3 (P = 0.003). Conclusions: Topical fluoroquinolones reach therapeutic levels in tears and cornea. Fluoroquinolones may be useful in treating bacterial keratitis even when there is an intact epithelium. Supported by a grant from the ACVO Vision for Animals Foundation. ABSTRACT NO.: 055 Comparison of two tonometers (TonoPen-VET and TonoPen-AVIA) in canine patients J. J. Studer,* R. D. Ring* and M. E. Payton† *The Animal Ophthalmology Clinic, Dallas, TX, USA, †Department of Statistics, Oklahoma State University, Stillwater, OK, USA Purpose: To compare intraocular pressure (IOP) values obtained with the TonoPen VET (TP) and the TonoPen AVIA (AV) applanation tonometers in clinically normal eyes of dogs. Methods: Sixty clinically normal dogs were enrolled in the study. There was no acclimation period prior to obtaining measurements. All dogs were manually restrained in a seated or sternal position. Two drops of topical anesthetic (proparacaine HCl) was instilled into each eye. Slit lamp biomicroscopy and indirect ophthalmoscopy were then performed on both eyes of each animal. If corneal pathology was noted, the animal was not included in the study. IOP was then measured using either the TP first (TP1 (30 animals)) or the AV first (AV1 (30 animals)). Within each group (TP1 and AV1), the IOP was measured first in the right eye (OD1) in half of the animals and first in the left eye (OS1) in half of the animals. Following IOP measurement with the first instrument and a waiting period of approximately 1 min, the IOP was measured with the second instrument in the same eye order that was performed with the first instrument. The data were analyzed using ANOVA procedures assuming a 2x2 factorial arrangement of treatments with side and first method as factors. Results: The mean IOP of both eyes for each of the four groups were as follows: 11.67 mm/Hg (TP1-OD1), 10.63 mm/Hg (TP1-OS1), 12.20 mm/Hg (AV1-OD1), and 11.38 mm/Hg (AV1-OS1). There was no instrument by side interaction (P = 0.7528), indicating that the order of procedures (instrument used or eye measured) performed was not statistically or clinically significant. The mean IOP of OD in all groups was 11.93 mm/Hg as compared to a mean IOP of 11.00 mm/Hg for OS. This is statistically significant (P = 0.0076). The mean IOP of all eyes measured with the TP was 11.15 mm/Hg and the mean IOP of all eyes measured with the AV was 11.79 mm/Hg. This value is not statistically significant (P = 0.0630). Conclusions: While there are minor differences, the two applanation tonometers are very similar in methodology, ease of use, consistency of performance and values obtained. The TonoPen-Vet produced values slightly lower than the TonoPen-Avia. This was neither statistically or clinically significant. While the values obtained in right eyes were statistically different than those obtained in left eyes, this is of little to no clinical significance. ABSTRACT NO.: 056 SOLX gold shunt implantation in dogs with primary glaucoma J. S. Sapienza,* J. Wolfer,† E. A. Lutz* and K. Kim* *Long Island Veterinary Specialists, Plainview, NY, USA, †Islington Animal Clinic, Toronto, Canada Purpose: To evaluate the postoperative clinical course of a novel glaucoma shunt, which channels aqueous humor from the anterior chamber to the space in dogs with primary glaucoma. Methods: A retrospective study was conducted on eight dogs (n = 9 eyes) with primary glaucoma that were implanted with a SOLX Gold shunt. This glaucoma implant is placed through a partial thickness incision into the anterior chamber and a penetrating incision posteriorly into the suprachoroidal space. The postoperative clinical course, intraocular pressure measurements, vision, and potential complications were evaluated. Results: Seven breeds of dogs were treated with the Gold shunt. One dog received bilateral gold shunt implants during the same operation. One dog had the original Gold shunt replaced with an upgraded model. Two neutered males, 1 intact male, 4 spayed and 1 intact female were treated. The right eye was affected in 4 cases, and the left eye in the remaining 5 cases. Four dogs were blind from glaucoma before surgery. One of these dogs had vision return after the intraocular pressure (IOP) was reduced after surgery. The shunts failed to control pressure adequately in 6/9 eyes. One patient with bilateral Gold implants remained normotensive and visual during the course of this study (4 months). Postoperative complications included poor IOP (n = 5 eyes), hyphema (n = 4), retinal detachment (n = 1), and loss of vision (n = 4). Conclusions: The SOLX Gold shunt can be easily implanted after several procedures, but failure to control the IOP resulted in poor IOP control in 6/9 eyes with loss of vision in 4/9 eyes. A larger series of patients with longer followup will be beneficial to further evaluate the long-term effects of this novel glaucoma shunt. ABSTRACT NO.: 057 Functional assessment and safety of irrigation with 5-Fluorouracil in a sealed-capsule irrigation device in the dog eye J. M. Histed, B. Nadelstein and R. V. English Animal Eye Care, Chesapeake, VA, USA Purpose: Pharmacologic modulation of lens epithelial cell (LEC) activity is an expanding area, limited by the ability to deliver agents to the desired location without collateral toxicity. Sealedcapsule irrigation is designed to allow targeting of LECs during cataract surgery and help prevent posterior capsule opacification (PCO). 5-Fluorouracil (5-FU) is a pyrimidine used to inhibit fibroblast proliferation and reduce postoperative scarring. The purpose of this study was to demonstrate the use of a sealed-capsule irrigation device in a clinical setting, as well as to study the safety of irrigation with 5-FU via application of such a system. Results will help guide future studies on the use of sealed-capsule irrigation in reduction of PCO. Methods: Six client-owned dogs undergoing bilateral phacoemulsification for cataracts were prospectively 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 398 a b s t r a c t s included in the study. One randomly selected eye received application of the sealed-capsule irrigation system and irrigation with 5-FU 50 mg/mL and the contralateral eye received standard cataract surgery and served as a control. Safety of 5-FU was evaluated by comparing aqueous flare, and corneal pachymetry findings preoperatively and 1, 1–10 days, 34 weeks, 23 months, and 6 months postoperatively. Photographs for PCO assessment were obtained at 2 weeks, 6 weeks, 3 months, and 6 months. Results: In all but one patient, the device could be safely applied to the capsule without damage. A good seal appeared to be formed with no apparent leakage detected in these patients. Mean corneal thickness in all control eyes (n = 6) post-operatively was 3.5% greater than preoperatively. In patients where a good seal was formed (n = 5) mean postoperative corneal thickness was 8.3% greater. When leakage of 5-FU occurred (n = 1) mean postoperative corneal thickness was 24.9% greater than preoperatively. There was no difference in aqueous flare values between control and study eyes in all patients (n = 5) where a good seal was formed. Aqueous flare was greater in patients (n = 1) where leakage of 5-FU occurred. Subjectively there is less PCO development in treated versus control eyes. Conclusions: A sealed-capsule irrigation system capable of inhibiting LECs and PCO is feasible in dogs and was safe with no clinically detectable collateral damage when no leakage occurs. However, subtle modification of the system could increase safety and usability in the canine eye. ABSTRACT NO.: 058 Efficacy and duration of proparacaine and tetracaine topical ophthalmic formulations on equine corneal sensitivity K. L. Sharrow-Reabe, W. M. Townsend and J. O. Campbell College of Veterinary Medicine, Michigan State University, East Lansing, MI, USA Purpose: This study compared the efficacy and duration of three different topical ophthalmic anesthetic formulations after application to the equine cornea. Methods: A masked study was undertaken on 20 normal eyes of 10 healthy horses. Each eye was randomly assigned to one of four groups (0.5% proparacaine hydrochloride solution, 0.5% tetracaine solution, 0.5% tetracaine gel, or saline control). Corneal anesthesia was measured by determining the corneal touch threshold (CTT) of each eye with a Cochet-Bonnet aesthesiometer. Each eye’s baseline CTT was recorded prior to anesthetic instillation at t = 0 min. The CTT values were recorded at t = 10 min and every 10 min until t = 60 min, then at t = 75 min and t = 90 min. Results: Mean baseline CTT was 1.25 +/)0.34 cm. The CTT of the control eyes did not differ from baseline. Maximum efficacy for each treatment occurred at t = 10 min. Tetracaine gel was the only treatment for which all eyes in the group reached a CTT of 0 cm. Duration of effect was 20 min for tetracaine solution and proparacaine solution and 30 min for tetracaine gel. Conclusion: Tetracaine gel demonstrated the greatest decrease in CTT and the longest duration of action. Proparacaine solution and tetracaine solution both decreased corneal sensitivity to a lesser degree and had a shorter duration of action. None. Tetravisc manufactured by Ocusoft of Richmond, TX. ABSTRACT NO.: 059 Periocular cutaneous mast cell tumors in cats: evaluation of surgical excision (33 cases, 1996–2009) K. W. Montgomery,* A. van der Woerdt,† S. M. Aquino,† J. S. Sapienza‡ and E. C. Ledbetter* *College of Veterinary Medicine, Cornell University, Ithaca, NY, USA, †The Animal Medical Center, ‡Long Island Veterinary Specialists, Plainview, NY, USA Purpose: To describe feline periocular cutaneous mast cell tumors (CMCT) and evaluate rates of local recurrence and metastases as well as survival time following surgical excision. Methods Medical records of 33 cats diagnosed with periocular CMCT were reviewed; cats were included if CMCT were surgically excised and the diagnosis was confirmed by histopathology. The appearance, size, location and histopathology findings of CMCT were evaluated. Rates of local recurrence, metastasis, and survival time following surgical excision were collected when available. Results: All periocular CMCT were restricted to the eyelids and were surgically excised; 3 cats were treated with adjunctive therapy (strontium-90 irradiation or cryotherapy) intraoperatively. Local tumor control was achieved in 21/22 cats with a minimum follow-up of 30 days (median follow-up time of 711 days); one cat developed disseminated CMCT but no local recurrence. Cats with periocular CMCT had a median survival time of 945 days. Metastatic disease involving peripheral lymph nodes or abdominal viscera was not detected in any cats at any time during the study. All periocular CMCT were classified as low-grade based on histopathology, and complete excision was achieved in approximately 50% of cases. Conclusions: Surgical excision of periocular CMCT in cats is an effective treatment option with rare local recurrence and metastases. ABSTRACT NO.: 060 Computed tomography evaluation of four retrobulbar injection techniques in the horse ABSTRACT NO.: 061 Retinal Pigment Epithelium (RPE) autotransplantation in porcine eyes I. Fernandez-Bueno,*,† J. M. Gonzalo-Orden,† M. RegueiroPurriños,† M. Parrado,* D. Hileeto,* E. Rodriguez de la Rua* and J. C. Pastor* *IOBA, University of Valladolid, Spain, †IBIOMED, University of León, Spain Prupose: RPE transplantation is considered a hopeful treatment for age-related macular degeneration (AMD). The purpose of this study is to improve a RPE autotransplantation technique, previously described by Van Meurs, in pig eyes, in order to use it as a control group in future stem cells transplantation studies. Methods: A total of 8 pigs (8 eyes) were operated on. In brief, standard 20G pars plana vitrectomy and lensectomy were performed using an Accurus Vitrectomy System (Alcon, TX). RPE graft was obtained after laser treatment (Iridex Corporation, CA) of the donor nasal retina by bimanual technique using vitreous forceps and scissors (Alcon/Grieshaber, Switzerland). RPE graft was then placed under the central retina using a Van Meurs’ forceps (D.O.R.C., France) through a retinotomy. Finally, eyes were filled with 1000 Cts. silicone oil (AJL, Spain). Ophthalmoscopic and anatomopathological studies were performed in control after the surgery and one month of follow up animals. Donor and transplant graft sites were evaluated in H&E and PAS ocular globe sections. GFAP and CRALBP antibodies were also used. Results: In 3 out of 8 cases surgery was not finished due to intraoperative complications. One case developed an acute endophthalmitis after one week. Control did not show differences compared to normal pig eyes. In the 3 cases followed up for a month the graft remained in place, but a proliferative vitreoretinopathy (PVR) developed. Histologic evaluation revealed a double RPE layer (native and graft) firmly attached, both layers demonstrating preserved cell polarity and uniform pigment distribution. Immunostaining revealed reactive gliosis in the PVR regions. Conclusions: RPE transplantation in porcine eyes shows promising results and remarkable potential, but the development of PVR is a major problem. Further studies are necessary in order to improve and use this model in research for AMD. Supported by Cajas de Ahorros de Castilla y León. ABSTRACT NO.: 062 Canine preiridal fibrovascular membranes: morphologic and immunohistochemical investigations M. K. Zarfoss,* C. B. Breaux,† H. E. Whiteley,* R. E. Hamor,* J. A. Flaws,* P. Labelle‡ and R. R. Dubielzig§ *University of Illinois College of Veterinary Medicine, Urbana, IL, USA, †WestVet Boise ID, ‡Antech Diagnostics Champaign IL, §University of Wisconsin Comparative Ocular Pathology Laboratory, Madison, WI, USA Purpose: Pathologic intraocular neovascularization is a key component of many canine ophthalmic diseases such as uveitis, retinal detachment, intraocular neoplasms, and corneal perforation. The purpose of this study was to evaluate the structure of preiridal fibrovascular membranes (PIFMs) associated with several different disease processes and to identify specific factors associated with their development in the canine eye. Methods: This study examined 36 enucleated canine eyes with the diagnosis of PIFM and one of the following: lens-induced uveitis, retinal detachment, iridociliary adenoma, corneal perforation, severe hyphema, or vitreal gliovascular membranes (canine ocular gliovascular syndrome, COGS). The following histologic/immunohistochemical stains were performed in all 36 PIFM eyes and four histologically normal eyes: hematoxylin and eosin, alcian blue periodic acid schiff (PAS), Masson’s trichrome, platelet endothelial cell adhesion molecule-1 (CD31), smooth muscle actin, vimentin, laminin, vascular endothelial growth factor (VEGF), and cyclooxygenase-2 (COX-2). Results: PIFM extracellular matrix staining was consistent with collagen and mucins in all cases and positive for laminin in most cases. All PIFMs contained CD31-positive vessels and predominantly lymphoplasmacytic inflammation. Both PIFM vessels and spindle cells were positive for laminin, vimentin, smooth muscle actin, VEGF, and COX-2. Secondary intraocular pathology and immunohistochemical staining of other intraocular structures are also reported. Conclusions: PIFM morphology and immunohistochemical characteristics were similar across six canine disease processes, suggesting analogous pathophysiologic mechanisms. COX-2 and VEGF were identified using immunohistochemistry and may play a role in PIFM development. Supported by ACVO Vision for Animals Foundation Grant 2007–8 and the Comparative Ocular Pathology Laboratory of Wisconsin. ABSTRACT NO.: 063 Limbal allograft transplantation in a dog with alkali-induced ulceration N. C. Whelan, S. G. Nykamp and C. L. Kerr Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada F. L. C. Brito,* O. Cunha† and J. L. Laus‡ *Universidade Federal Rural de Pernambuco – Unidade Acadêmica de Garanhuns, Pernambuco, Brazil, †Universidade Federal do Paraná – Campus de Palotina, Palotina, Brazil, ‡Faculdade de Ciências Agrárias e Veterinárias, UNESP, Jaboticabal, Departamento de Clı́nica e Cirurgia Veterinária, São Paulo, Brazil Purpose: To compare four retrobulbar injection techniques (4-point retrobulbar, lateral above (AZA) or below the zygomatic arch (BZA) and vertical to the posterior cranial orbital rim (VPCOR)) using CT. Methods: Thirty orbits from previously frozen equine heads were randomly allocated to one of four injection techniques. Heads were placed in ventral recumbency and 16G Angiocatheter(s) were placed. Injections were made with a 50:50 mixture of 1% methylene blue and iohexol. For the 4-point retrobulbar injection, 10 ml was injected/site. For the remaining three injection techniques, 15 ml were used. Images were reviewed for needle location relative to the orbital fissure and intraconal accuracy. Results: For the 4-point block, the medial and ventral needle tips were closest to the optic fissure. Of the other three techniques, the mean needle tip distance from the orbital fissure was AZA 19.9, BZA 16.3, and VPCOR 31.4mm). With the 4-point technique, intraconal accuracy was best with medial placement of the needle (4/9), whilst the dorsal and lateral needle locations were extraconal 7/9 times. For the other three techniques, the needle was intraconal 7/7, 6/7 and 5/7 times for the AZA, BZA and VPCOR respectively. Conclusions: AZA and BZA provided the closest proximity to the orbital fissure. The AZA, BZA or VPCO techniques all provided good intraconal deposition of material. Supported by the Ontario Veterinary College, University of Guelph. Purpose: We report here the case of a 3-month-old female mongrel dog with a history of apathy and previous contact with an alkaline cleaning product (sodium hydroxide). Methods: The dog was reluctant to open the right eye. Ophthalmic examination revealed blepharospasm, photophobia, epiphora, discrete chemosis, conjunctival hyperemia, and diffuse corneal edema involving the limbus. The fluorescein test was positive and the result of the Schirmer tear test was 32 mm/min. No signs of alterations were observed in the left eye. Results: On the basis of these findings the diagnosis was alkali-induced ulceration and limbal allograft transplantation was performed. Corneal vascularization was observed by the third postoperative day, with intensification in vessel number and caliber on subsequent days and small areas of corneal transparency. On postoperative day 21, vessels were still present in moderate numbers and small areas of corneal transparency were observed close to the transplant. On day 30 and 45, the cornea presented central vascularization and very discrete and poorly significant areas of transparency were observed close to the transplanted limbus. Conclusion: The present results show that limbal allograft transplantation is a feasible procedure for the therapeutic management of alkali-induced corneal ulcers, since it prevents large-scale conjunctivalization and permits to safe the eye. 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 abstracts 399 ABSTRACT NO.: 064 Effects of continuous infusion of dopamine or dobutamine in intraocular pressure in association with cardiovascular parameters in dogs anesthetized with isoflurane F. L. C. Brito,* M. G. Sousa,† R. Carareto,† A. B. De Nardi,† N. Nunes‡ and J. L. Laus‡ *Universidade Federal Rural de Pernambuco – Unidade Acadêmica de Garanhuns, Pernambuco, Brazil, †Universidade Federal do Tocantins – UFT, Campus of Araguaı́na, Tocantins, Brazil, ‡São Paulo State University – UNESP, Department of Veterinary Medicine and Surgery, College of Agricultural and Veterinary Sciences, Campus of Jaboticabal, São Paulo, Brazil Purpose: The objective of this study was to determine the intraocular pressure (IOP) and cardiac changes in dogs anesthetized with isoflurane and treated by continuous infusion of dopamine or dobutamine. Methods: We used 16 healthy dogs of various breeds and ages, males or females with average weight of 11 kg. The animals were divided into 2 groups of 8 animals for group. Both groups were anesthetized with isoflurane (1.0 MAC). Group 1 was subjected to continuous infusion of dopamine (5 lm.kg-1.min-1) and group 2 the infusion of dobutamine (5 lm.kg-1.min-1). In both groups the IOP was measured by using applanation tonometry. Were also recorded mean arterial pressure (MAP), systolic blood pressure (SBP), diastolic blood pressure (DBP), central venous pressure (CVP) and perfusion pressure (PP) in the eye. The parameters were measured in time: before the infusion anesthesia (M0), after stabilization anesthesia (M1) and every 10 min of infusion (M2, M3 and M4). Data analysis was used analysis of variance (ANOVA) followed by Tukey test. Results: There was no significant difference in IOP before and after infusion of drugs over time. But change occurred compared the two groups. There was a marked decrease in ocular perfusion pressure. The cardiovascular parameters did not differ between groups. Conclusion: Care must be taken regarding the use of dopamine or dobutamine in patients with ocular hypertension. ABSTRACT NO.: 065 Cataracts in 44 dogs (77 eyes): a comparison of outcomes for no treatment, topical medical management, or phacoemulsification with intraocular lens implantation C. C. Lim,* S. Bakker,* C. L. Waldner,† L. S. Sandmeyer* and B. H. Grahn* *Department of Small Animal Clinical Sciences, †Department of Large Animal Clinical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Canada Purpose: The primary objective was to compare outcomes for patients with cataracts that either received no therapy, topical medical therapy only, or phacoemulsification with intraocular lens implantation. The second objective was to document the most common complications associated with treatment failure. Methods: Medical records of dogs diagnosed with cataracts at the Western College of Veterinary Medicine were reviewed retrospectively. Patients were divided into 3 groups: (1) patients that did not receive any treatment for cataracts, (2) patients that received topical medical treatment only, and (3) patients that underwent phacoemulsification with intraocular lens implantation. Occurrence of treatment failure was compared using the Cox proportional hazards model. Results: Median follow up time for all patients was 853 days. Failure occurred in all untreated cases (8 of 8 eyes) and was 64.5 and 255 times more likely than failure in medically and surgically treated eyes, respectively. Failure was 4 times higher in dogs receiving medical treatment (20 of 35 eyes) only when compared to dogs undergoing phacoemulsification with intraocular lens implantation (7 of 34 eyes). For both medically and surgically-treated groups, success rate for mature and hypermature cataracts was less than for immature cataracts. However, regardless of cataract stage, the chance of success was still higher for eyes undergoing phacoemulsification than for eyes that received medical management only. The most common complications associated with failure in medically treated patients were cataract progression, postoperative uveitis, postoperative corneal disease, and glaucoma. The most common complications associated with surgical failure were postoperative corneal disease, intraoperative hyphema, and glaucoma. Conclusions: Surgery has a superior success rate when compared to other modes of cataract management, especially when performed early in the disease process. Lack of medical or surgical therapy results in a high rate of complications that often results in loss of the eye. ABSTRACT NO.: 066 Association between dog leukocyte antigen haplotype and golden retriever pigmentary uveitis W. M. Townsend,* A. Mankey† and J. A. Gerlach‡ *College of Veterinary Medicine, Michigan State University, East Lansing, MI, USA, †College of Human Medicine, Michigan State University, East Lansing, MI, USA, ‡Natural Sciences, Department of Physiology and Human Pathology, Michigan State University, East Lansing, MI, USA Purpose: To determine if an individual’s dog leukocyte antigen (DLA) haplotype correlated with the development of Golden Retriever pigmentary uveitis. Methods: The pedigrees of and blood samples from Golden Retrievers diagnosed by ACVO diplomats as affected with pigmentary uveitis were collected. Blood samples were collected from their unaffected first and second degree relatives as well. Pedigrees from 220 animals were analyzed to determine the mode of inheritance of pigmentary uveitis. Genomic DNA was isolated from peripheral blood by commercial column methodologies. The DLA*DQB locus was amplified and the product diluted and cycle sequenced using a chain termination method. Sequences from 65 animals were compared for allele and sequence similarity and dissimilarity. Results: No association could be detected between the DLA*DQB locus sequence and the development of pigmentary uveitis. The mode of inheritance cannot be conclusively determined and is either autosomal dominant with incomplete penetrance or a recessive trait. Conclusion: Golden Retriever pigmentary uveitis does appear to be an inherited trait. The DLA*DQB locus does not appear to play a role in the development of pigmentary uveitis. Michigan State University Companion Animal Fund. ABSTRACT NO.: 067 Immunohistochemical evaluation of fibrovascular and endothelial-like pre-iridal membranes in dogs B. S. Bauer,* L. S. Sandmeyer,* R. B. Hall† and B. H. Grahn* *Department of Small Animal Clinical Studies, Western College of Veterinary Medicine, Saskatoon, Saskatchewan, Canada †College of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada Purpose: Histologically, two different types of pre-iridal membranes appear to occur in diseased canine globes: fibrovascular and endothelial-like. It has been proposed that the endothelial pre-iridal membrane in iridocorneal endothelial (ICE) syndrome in humans arises through metaplastic transformation of endothelial cells into epithelial-like cells. The purpose of this study is to (1) evaluate immunohistochemical staining of the two types of pre-iridal membranes in dogs to determine if endothelial cell metaplasia plays a role in membrane formation and (2) determine if the ocular disease correlates with the type of membrane present. Methods: H&E stained slides of 28 enucleated canine specimens with pre-iridal membranes were examined. The globes were divided into two groups based on the appearance of the pre-iridal membrane: fibrovascular or endothelial-like, and the histopathologic diagnosis was recorded. Immunohistochemical labeling with antibodies to vimentin, cytokeratin AE1/AE3 and Factor VIII related antigen was completed on the globes. The histopathologic diagnoses were compared between the two groups. Results: The fibrovascular and endothelial-like membranes stained positive for vimentin and negative for cytokeratin AE1/AE3. All fibrovascular membranes stained positive for Factor VIII related antigen compared to the endothelial-like membranes which stained negative. The endothelial-like membrane appeared more commonly in globes with primary glaucoma. Conclusions: Immunohistochemistry does not support the hypothesis of metaplastic transformation of endothelial cells into epithelial-like cells in the canine globes with endothelial-like pre-iridal membranes. The increased frequency of endothelial-like membranes in primary glaucoma may indicate a role of iridocorneal endothelialization in the pathogenesis of the condition. Supported by the University of Saskatchewan Diode Laser Fund. ABSTRACT NO.: 069 Dacryocystitis associated with cutaneous erosion in a puppy C. F. Campos*,† and M. D. Barros* *Veterinária Ypiranga, †Centro de Criação de Animais de Laboratório, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil Purpose: This study aims to document a case of recurrent dacryocystitis with cutaneous involvement. Methods: A 60-day-old female Yorkshire was referred for ophthalmic evaluation due to recurrent purulent eye discharge. Complete ophthalmic examination revealed abundant purulent ocular discharge emerging from mildly dilated lachrymal puncta. The puppy did not present any other health problems and vaccinations and worm preventions were current. Dacryocystorhinography did not reveal the presence of foreign bodies. The puppy received ciprofloxacin ointment for 20 days, the signs ceased but recurred and after 2 days there was also bilateral purulent discharge from the nares. The lachrymal puncta were cannulated and flushed. After 5 days of the beginning of the nasal discharge the dog also presented a perinasal vesicle, which ruptured, leading to a cutaneous erosion. Samples for impression cytology and microbial culture/antibiogram tests were collected from the perinasal skin and ocular discharge respectively. Ocular moxifloxacin and chlorexidin were used, with temporary regression of the ocular and skin signs. Oral doxiciclin was prescribed but the owners did not administer. When the treatment stopped ocular and nasal discharges were observed again and new nare skin lesions were found. The third eyelids and inferior eyelids margins also ulcerated. A cutaneous sample from the nare was collected for biopsy. The lachrymal puncta were frequently cannulated and flushed during treatment. Treatment with ocular moxifloxacin and chlorexidin and oral doxiciclin was prolonged for one month and the ocular and nasal discharges ceased. Results: No anaerobic microorganisms neither fungal components were isolated. Staphylococcus epidermidis was isolated from the ocular discharge. The skin lesions cytology and histopathology revealed a suppurative inflammatory response. Skin lesions occurred only when there was discharge from the nares. After 30 days of oral doxiciclin and topical ciprofloxacin and chlorexidin the discharge ceased and skin lesions scarred. Conclusions: Considering that perinasal skin lesions occurred only when there was evident discharge from the nares, it was possibly associated with toxins produced by Staphylococcus epidermidis, as happens in the scalded skin syndrome in children and babies. Dacryocystitis may require prolonged treatment. ABSTRACT NO.: 070 Cartilage reversal: a technique for correction of everted nictitans E. D. Wolf Southern Eye Clinic for Animals, Tampa, FL, USA Purpose: Develop a technique for surgical correction of eversion of the nictitans cartilage that restores normal contact between the cornea and the advancing margin of the nictitans while maintaining full excursion of the nictitans across the cornea. Methods: Eight eyes of five canine patients with everted cartilage of the nictitans underwent surgery to reverse the everted portion of the vertical stem portion of the T-cartilage of the nictitans. Results: A total of 7/8 operated eyes resulted in normal contact between the leading margin of the nictitans and the cornea while maintaining full excursion across the cornea as demonstrated by retropulsion. Conclusion: A new technique is developed to reverse a portion of the cartilage of the third eyelid to correct eversion of the nictitans while maintaining normal nictitating excursion across the cornea. ABSTRACT NO.: 071 Use of a buccal mucosal island graft for surgical repair of feline eyelid agenesis A. Welihozkiy, S. Pizziran and C. G. Pirie Tufts Cummings School of Veterinary Medicine, Grafton, MA, USA Purpose: To describe a novel surgical technique combining a buccal mucosal island graft with a sliding skin graft for the repair of feline upper eyelid agenesis. Methods: Upper eyelid agenesis in conjunction with superficial keratitis and blepharospasm was diagnosed in three domestic short hair cats (two bilateral). Peters’ anomaly was noted in two patients and PPM’s were present in one patient. Repair of the defect was accomplished using a buccal mucosal island graft in 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 400 a b s t r a c t s conjunction with a single pedicle advancement flap from the dorso-lateral skin of the forehead. The oral graft was harvested from the upper vestibular mucosa. The created defect in the buccal mucosa was left to heal by secondary intention. In one lid, a concurrent Stades-procedure was performed. Results: Median follow-up time was 217 days. All cats remained comfortable throughout follow up-period. No island graft rejection was noticed. Superficial keratitis improved in all cats. A rudimental but anatomically present upper conjunctival fornix was created and was functional during blinking in all patients. Mild trichiasis in conjunction with occasional mild blepharospasm was noted in two eyes; in one lid further application of cryosurgery was required. Conclusion: The presented method has the advantage of creating a functional dorsal fornix lined by mucosal membrane. This reflects in a better motility of the upper lateral lid during blinking and protects the dorso-lateral cornea from exposure and trichiasis. This technique may represent a valuable alternate surgical approach to the previously described surgical methods for feline eyelid agenesis repair. ABSTRACT NO.: 072 Sub-choroidal foreign body in a dog – a case report A. Welihozkiy, S. Pizzirani and C. G. Pirie Tufts Cummings School of Veterinary Medicine, Grafton, MA, USA Purpose: To describe a case of a sub-choroidal foreign body in a dog. Methods: A 2-year-old MC Springer Spaniel was referred for a 3 week history of exophthalmos, periorbital swelling, and chemosis associated with fundic changes OD. On examination, slight chemosis and elevation of the nictitating membrane were noted. Exophthalmos was questionable. Retropulsion was normal OU. No pain was elicited in opening of the mouth. Vision and ocular reflexes were normal OU. Indirect ophthalmoscopy OD revealed an approximately 8 mm peripapillary hyporeflective tapetal lesion with subretinal edema. Contrast enhanced CT showed a focal, poorly defined hyperdense tissue along the caudomedial aspect of the globe OD, without orbital involvement. Ultrasound showed a single, ovoid nodule, with a hypoechoic center within the posterior wall of the globe. Focal retinal separation was evident. Based on these findings, a guarded prognosis for the globe and a main differential diagnosis being neoplastic disease, enucleation OD was performed. Results: Histopathology of the globe revealed focal thickening of the posterior wall by a proliferation of neutrophils, macrophages, lymphocytes and plasma cells. These cells surrounded a focus of foreign material most consistent with plant material located between the sclera and the choroid. Severe choroidal inflammation was evident. Regional retinal detachment with hypertrophy of the RPE layer was noted. No evidence of retrobulbar/ orbital neoplasia was found in examined sections. Conclusions: This report describes a rare case of subretinal migration of plant material and outlines the difficulty of an etiological diagnosis using ocular imaging. ABSTRACT NO.: 073 A retrospective review of clinical ocular and orbital magnetic resonance imaging (MRI) in canine and feline patients (2004–2009) M. Armour,* G. Dell’Anna,† D. Esson* and M. Broome† *Eye Care for Animals, Tustin, CA, USA, †Advanced Veterinary Medical Imaging, Tustin, CA, USA Purpose: To review the distribution of ocular and orbital disease in canine and feline patients undergoing MRI. Specifically, the roentgen characteristics of predominant tumor types are described. Findings in patients diagnosed with presumptive optic neuritis are also described. Methods: Records of 78 patients that underwent MRI following presentation for signs of ocular/orbital disease were reviewed. Results: The most common tumor types diagnosed by MRI-guided biopsy were carcinoma (11), sarcoma (9) and lymphoma (6). Orbital carcinomas were typically characterized by irregular margination and intracranial/sinonasal extension. The majority of sarcomas involved unilateral sinonasal extension; however, intracranial extension was present in only 50%. Orbital lymphomas were generally unilateral and demonstrated no intracranial/sinonasal extension or associated osteolysis. Intracranial masses consistent with meningiomas were identified in nine cases. A diagnosis of inflammatory (non-neoplastic) orbital disease was made in 11 cases. A diagnosis of presumptive optic neuritis was made in 11 patients following adjunctive diagnostic testing including CBC, biochemistry, infectious disease titers and electroretinography where appropriate, as well as cerebral spinal fluid (CSF) analysis in all 11 patients. CSF analysis was unremarkable in 9/11, and demonstrated changes in 2/11. Normal MRI scans were noted in 3/11. Diffuse enhancement of one or both optic nerves and/or the optic chiasm following the administration of gadolinium-based contrast medium was noted in 2/11. Multifocal, patchy contrast enhancement of the cerebral cortex was noted in 6/11. Conclusions: Essential (MRI) roentgen characteristics may be helpful when determining the nature and prognosis of ocular/orbital disease in canine and feline patients. Funding: N/A. ABSTRACT NO.: 074 Ophthalmic examination findings in a captive mob of western gray kangaroos (Macropus fuliginosus) A. L. Labelle,* M. Low,* R. E. Hamor,* C. B. Breaux,* J. N. Langan,*,† M. K. Zarfoss* and T. T. Zachariah‡ *College of Veterinary Medicine, University of Illinois, IL, USA, †Chicago Zoological Society’s Brookfield Zoo, ‡Chicago Zoological and Aquatic Animal Residency Program, College of Veterinary Medicine, University of Illinois, IL, USA Purpose: To describe the ophthalmic examination findings in a captive mob of western gray kangaroos (Macropus fuliginosus). Methods: Complete ophthalmic examination of eight adult western gray kangaroos was performed under chemical restraint. Examination included intraocular pressure measurement by rebound and applanation tonometry, fluorescein staining, corneal diameter measurement, slitlamp biomicroscopy, and indirect funduscopy. Results: Ophthalmic abnormalities were documented in 4/8 kangaroos (50%) and included a lower lid notch defect, subepithelial corneal fibrosis, cataracts, nuclear sclerosis, vitreal degeneration, an optic nerve coloboma, and choroidal hypoplasia. The corneal diameters had a mean of 19.52 mm, SD of 2.16 mm and 95% CI of 18.71–20.32 mm. Intraocular pressure (IOP) as estimated by rebound tonometry had a median of 9.00 mmHg, a 25–75% quartile range of 6.50–10.75 mmHg, and a minimum–maximum range of 5.00–23.00 mmHg. Intraocular pressure as estimated by applanation tonometry had a median of 11.50 mmHg, a 25–75% quartile range of 10.00–17.00 mmHg, and a minimum-maximum range of 9.00–20.00 mmHg. The applanation tonometry method frequently produced a larger value for IOP than the rebound method. Correlation between the two tonometry methods improved when only one clinician performed the measurements. This trend was also seen with the level of agreement between the two methods of tonometry, which was assessed as fair using Bland-Altman analysis. Conclusions: This is the first report of ocular examination findings in western gray kangaroos and provides ranges and values for tonometry and corneal diameter. ABSTRACT NO.: 075 Standard full-field electroretinogram in diabetic dogs with cataracts A. M. V. Safatle,* A. P. Havenegaard,* D. Otsuki,† T. L. Martins,† M. Kahvegian,† A. Berezovsky‡, S. R. Salomão‡ and P. S. M. Barros* *Laboratory of Investigation in Comparative Ophthalmology, School of Veterinary Medicine, University of São Paulo, São Paulo, Brazil, †School of Medicine, University of São Paulo, São Paulo, Brazil, ‡School of Medicine, Federal University of São Paulo, São Paulo, Brazil Purpose: To evaluate standard full-field electroretinography in diabetic dogs presenting mature or hypermature cataracts. Methods: Standard full-field electroretinogram recordings were obtained from seventy-four dogs, with ages varying from 6 to 13 (mean = 9.4 years-old; SD = 1.97) allocated in three groups: NG – normal dogs; CG – dogs presenting cataracts and DG – diabetic dogs presenting cataracts. Amplitude (microvolts) and implicit time (ms) were determined for each of the five standard full-field ERG responses (rod response, maximal response, oscillatory potentials, single-flash cone response and 30 Hz flicker). Results: Oscillatory potentials amplitudes were significantly lower in diabetic dogs when compared with normal dogs (7.4 lV versus 9.9 lV; P = 0.033).Flicker amplitudes were significantly reduced when comparing DG with NG (7.2 lV versus 15.3 lV; P = 0.000).There were prolonged implicit times in diabetic animals, when compared with normal and cataract groups. This difference was statistically significant comparing NG with DG for rod responses(48.2 ms versus 56.5 ms; P = 0.000), maximum responses (29.9 ms versus 31.9 ms; P < 0.05), cone responses (28.8 ms versus 30.7 ms; P < 0.05) and 30 Hz flicker responses (21.9 ms versus 24.1 ms; P < 0.05). Conclusions: Standard full-field ERG changes were found in diabetic dogs, such as amplitude reduction in oscillatory potentials and flicker response and prolonged implicit time in rod, maximal, cones and flicker response. These data suggest standard full-field ERG may be useful to investigate retinal dysfunction in dogs, such as diabetic retinopathy, when fundoscopic examination cannot be performed. Supported by FAPESP grant 02/08051-9 and CNPq grant 501494/2005-7. ABSTRACT NO.: 076 Comparative effects of Nepafenac 0.1% and Dexametasone 0.1% eye drops after experimental phacoemulsification L. F. M. Barros,* E. Mattos jr,* R. L. Almeida,† D. P. Rivelli,† S. B. M. Barros,† P. S. M. Barros* and A. J. Stopiglia* *School of Veterinary Medicine/University of São Paulo, São Paulo, Brazil, †School of Pharmaceutical Sciences/University of São Paulo, São Paulo, Brazil Purpose: To compare the efficacy of Nepafenac 0.1% and Dexametasone 0.1% to control inflammation after experimental phacoemulsification in rabbits. Methods: Seventeen rabbits were submitted to phacoemulsification in one eye. They were divided into three groups: Control Group (CG; n = 5) -moxifloxacin 0.5% (Alcon Laboratorios do Brasil, São Paulo, Brazil) 4 times a day; Nepafenac Group (NG; n = 6) -nepafenac 0.1% (Alcon Laboratorios do Brasil, São Paulo, Brazil) and moxifloxacin 0.5% 4 times a day and Dexametasone Group (DG; n = 6) dexametasone 0.1% (Alcon Laboratorios do Brasil, São Paulo, Brazil) and moxifloxacin 0.5% 4 times a day. Aqueous humor samples were obtained immediately before surgery and at days 1, 2, 3, 7 and 15 after surgery to access protein and ascorbic acid levels. Central corneal pachimetry (CCP) and intra-ocular pressure (IOP) were evaluated as well as clinical parameters (blepharospasm, conjuctival discharge, hyperemia, corneal edema and aqueous flare) were scored (0 = normal, 1 = mild, 2 = moderate, 3 = severe). Results: No differences in blepharospasm, conjuctival discharge and flare were observed between groups. DG showed higher scores of conjunctival hyperemia when compared with NG only at day 7 (P = 0.016). NG presented lower scores of corneal edema at days 2, 3, 7 and 15 (P = 0.004; 0.034; 0.035 and 0.062 respectively). IOP values of DG (36.83 ± 4.61 mmHg) were higher than NG and CG (24.17 ± 4.95 mmHg and 19.60 ± 1.29 mmHg) only at day 1 (P = 0.031). Lower values of CCP were observed in NG at all days after surgery but with no statistical significance. Regarding ascorbic acid levels, we observed a decrease at day 1 in all groups; 482.39 ± 69.54 lM; 409.47 ± 24.43 lM and 461.52 ± 66.98 lM (DG; NG; CG). While NG levels increased during post op time, DG levels remained decreasing until day 15 (593 ± 28.00 lM and 208 ± 30.81 lM; P = 0.002). Protein levels of NG were discreetly lowers than DG in all moments but only in day 2 higher concentration were observed when compared to DG (9.59 ± 1.65 g/L and 5.27 ± 0.63 g/L). Conclusion: Nepafenac treatment was more efficient than Dexametasone in the recover of aqueous humor ascorbic acid levels and reducing protein levels. Nepafenac seems to be efficient in reducing CCP, controlling IOP and inflammatory clinical signs when compared to Dexametasone. Supported by CAPES (PhD fellowship of Luiz Felipe de Moraes Barros). ABSTRACT NO.: 077 A survey of concurrent findings in canine and feline patients presenting for hyphema at a private referal clinic in Michigan from 2004 to 2008 C. D. Bliss, G. H. Sila, R. J. Morreale and D. W. Lorimer Michigan Veterinary Specialists, Department of Ophthalmology, Southfield, MI, USA Purpose: To evaluate clinical cases presented for hyphema at a private referral practice regarding signalment, examination findings, concurrent diagnoses, and outcome. Methods: Medical records of canine and feline cases presenting to Michigan Veterinary Specialists from 2004 to 2008 were reviewed (general population). Cases were included if there was a diagnosis of hyphema and a complete ophthalmic examination (study population). Populations were compared using paired t-test and a Chi-square contingency table. Results: Thirty-eight canines and six felines were included with 45 eyes affected (26 left, 19 right). Median age was 10.5 years (range 8 months to 20 years); 24 were male; 20 were female; breed varied. Thirty-five eyes had total 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 abstracts 401 hyphema (obscuring visualization of intraocular structures); 10 had partial hyphema. Median IOP was 15 mmHg (range 4 mmHg to 59 mmHg), median STT value was 23 mm/minute (range 4 mm/minute to 35 mm/minute) and two eyes had positive fluorescein staining. Concurrent diagnoses included retinal disease (20%), uveitis (15%), cataracts (14%), glaucoma, (11%), intraocular/extraocular mass (10%), concussive/perforating trauma (9%), history of ocular surgery (6%), systemic hypertension (4%), keratoconjunctivitis sicca (4%), lens luxation (2%), corneal ulcer (2%), and multiple congenital anomalies (1%). Ninety-two percent were considered non-visual at time of last evaluation. Conclusions: Median age of the study population was statistically higher than the general population with Labrador and Golden Retrievers overrepresented in the study. Most affected eyes had total hyphema and the most common concurrent diagnoses were retinal detachment (20%), uveitis (15%), and cataracts (14%). The prognosis for vision in cases with hyphema was poor. ABSTRACT NO.: 078 Effect of eyelid manipulation and manual jugular compression on intraocular pressure measurement in dogs H. E. Klein,* S. G. Krohne,* J. Stiles* and A. S. Mohamed† *Department of Veterinary Clinical Sciences, †Department of Comparative Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, IN, USA Purpose: To determine the effect of eyelid manipulation and manual jugular compression on intraocular pressure (IOP) measurement by applanation tonometry (Tono-Pen) in normal dogs. Methods: IOP was measured in 57 eyes of 30 dogs using six methods of eyelid manipulation and/or jugular compression. The methods used in each eye included (a) minimal eyelid manipulation, (b) maximal dorsoventral extension of the eyelids, (c) lateral eyelid extension, (d) manual compression of the ipsilateral jugular vein (e) manual compression of both jugular veins, and (f) lateral eyelid extension and manual compression of both jugular veins. Significant difference between each method was assessed by GLM and Tukey’s studentized range test. Results: The two manipulations that caused the greatest mean increase in IOP were lateral eyelid extension combined with compression of both jugular veins (17.6 mmHg increase) and lateral extension alone (16.5 mmHg increase). These results were statistically significant (95% CI = 15.7–19.6 and 95% CI = 14.5–18.4 respectively). Lateral or dorsoventral eyelid extension also significantly increased mean IOP. Compression of the ipsilateral jugular vein did not significantly alter IOP compared to minimal eyelid manipulation. Conclusions: Traction on the eyelids and/or pressure on both jugular veins can significantly and clinically increase IOP values as measured by the Tono-pen in normal dogs. ABSTRACT NO.: 079 Canine optic neuritis: retrospective analysis of 20 cases A. J. Specht, M. L. Busch and C. E. Plummer College of Veterinary Medicine, University of Florida, Florida, USA Purpose: To review common clinical presentations, underlying causes, treatments, and prognosis for visual recovery of canine optic neuritis cases. Methods: Retrospective analysis of medical records of 20 dogs diagnosed with optic neuritis between December 1996 and September 2007. Information about signalment, medical history, physical and ophthalmologic examination, diagnostic testing, treatments, and ultimate visual status were recorded. Results: Bilateral (15, 75%) presentation was more common than unilateral (5, 25%) and papillar neuritis (14/20, 70%) was more frequent than retrobulbar neuritis (6/20, 30%). Within the entire study population, three dogs were diagnosed with an infectious agent, five dogs had meningitis or menigoencephalitis, one dog had confirmed neoplastic disease, and 11 dogs had optic neuritis of unknown etiology due to lack of substantive diagnostic testing. Six of 14 (43%) dogs with follow-up information available demonstrated visual improvement after receiving corticosteroids. Though not statistically significant, a trend towards better visual outcome was seen in dogs with unilateral disease, retrobulbar disease, or with known potential infectious etiologies. Conclusions: Serologic testing and advanced imaging of the head appeared more beneficial than basic biochemical and hematologic analysis or general imaging for identifying possible underlying disease. It was not possible to determine whether corticosteroid therapy significantly altered prognosis, but there did not appear to be an association between dosage of corticosteroid received and visual outcome. ABSTRACT NO.: 080 Evaluation of precorneal tear film pH in clinically healthy dogs R. A. Armentano, A. J. Specht, C. E. Plummer, A. E. S. Stone and J. A. Wuerz Veterinary Medical Center, University of Florida, Florida, USA Purpose: To establish a range of precorneal tear film (PTF) pH measurements in a population of clinically healthy dogs as well as to determine the degree of correlation between PTF pH and pH of venous blood. Methods: This study was designed for prospective, sequential enrollment of clinically healthy canines (determined by history and physical examination) admitted to the Univeristy of Florida Veterinary Medical Center for wellness examination. Precorneal tear fluid samples were taken from the lower conconjuntival fornix of both eyes in randomized order with a surgical eye spear (sponge). A venous blood sample was also taken for pH measurement. Samples were analyzed immediately after collection with a silicone chip pH meter (IQ Scientific Instruments, model IQ125) which was calibrated prior to every measurement. Results: Mean PTF pH was 7.5 with a reference range (mean ± 2 standard deviations) of 6.4 to 8.6. The minimum measured value was 5.5, and maximum was 9.1. Mean measured venous blood pH was 7.6, with a smaller standard deviation of (±0.2). Measured values for PTF pH and blood pH were not significantly correlated. Conclusions: There appears to be substantial variation in measured pH of PTF from clinically healthy dogs. In addition, PTF pH does not appear to be a passive reflection of blood pH. Further study is necessary to elucidate factors that influence tear pH in canines. ABSTRACT NO.: 081 In vitro culture and reconstruction of equine corneolimbal epithelial cells on amniotic membrane C. E. Plummer, J. Coleman and H. Sapp College of Veterinary Medicine, University of Florida, Gainesville, FL, USA Purpose: To describe a procedure for harvesting and expanding equine limbal epithelial cells (LEC) in vitro and for constructing sheets of epithelium upon amniotic membrane (AM). Methods: Equine corneolimbal explants were harvested from cadavers and expanded in cell culture. The resultant LEC were placed upon AM and examined for proliferation and expansion. Results: LEC are easily obtained from limbal biopsies and are amenable to proliferation in culture. When placed upon pieces of preserved AM, these cells form sheets and orient themselves into an organized epithelial construct. Conclusions: LEC act as progenitor cells for the proliferation and healing of corneal epithelium. AM as a substrate for cultured LEC has tremendous potential for the advancement of therapies aimed at ocular surface reconstruction. ABSTRACT NO.: 082 Anesthetic induction with propofol and (S) ketamine for routine ERG in cataractous dogs H. D. Parchen, M. Bacellar, F. Montiani-Ferreira and R. G. D.’OC Vilani Universidade Federal do Paraná, Curitiba, Paraná, Brazil Purpose: To investigate the necessary dose and describe the characteristics of anesthesia with the combination propofol and (S) ketamine to perform routine electroretinographic procedures in dogs. Methods: Electroretinography (ERG) was performed in 12 ASA III or IV cataractous dogs, of different breeds and ages (varying from 6 to 10 years of age). Dogs were premedicated intravenously with diazepam (0.5 mg/kg, Compaz, Cristália, São Paulo-SP, Brazil) and morphine (0.5 mg/kg, Dimorf, Cristália, São Paulo-SP, Brazil). Fifteen minutes later, propofol (1.5 mg/kg/min, Propovan, Cristália, São Paulo-SP, Brazil) and (S) ketamine (0.5 mg/kg/min, Ketamin-S(+), Cristália, São Paulo-SP, Brazil) were infused together using 1 ml seringes until the dogs permitted positioning of blepharostat, corneal and subcutaneous ERG electrodes without any reaction. A short flash ERG protocol that uses high light intensities single flashes (3000 and 10 000 mcd.s/m2) for stimulation (HMsERG, RetVetCorp, Inc., Columbia, Missouri) was performed. Besides the time of induction, the duration of akinesia of the eyelids and the time to complete recovery, the following physiologic parameters were controlled: cardiac rate, respiratory rate, rectal temperature and blood oxygen saturation. Statistical analysis to compare the physiologic parameters was performed using ANOVA. Results: The mean of induction time was 125.5 ± 56.29 seconds, resulting in a mean dose of 3.13 ± 1.4 mg/kg of propofol and 1.04 ± 0.46 mg/kg of (S) ketamine. The best feature of this anesthetic protocol was that it was able to produce akinesia and centralization of the globe, for the entire length of the ERG. With this dose, the mean duration of akinesia of the eyelids to perform the ERG was 28 ± 10.59 minutes and complete recovery time occurred at 39 ± 4.5 minutes after the anesthetic induction. No statistical differences in any physiologic parameters were observed analyzed when compared with the values before premedication. One dog, however, presented reduction of SpO2 to 85% after the anesthetic induction and needed to receive 100% O2 by facial mask. In all the animals rectal temperature reduced from 38.47 ± 0.96 to 37.67 ± 0.70C at the end of the anesthesia, but this trend was not statistically significant. Conclusions: The infusion of 3.13 ± 1.4 mg/kg of propofol and 1.04 ± 0.46 mg/kg of (S) ketamine, during approximately 125 seconds, promotes a relatively safe anesthesia even for older dogs with adequate centralization of the globe, with the benefit of producing eyelid and globe akinesia to perform ERGs. ABSTRACT NO.: 083 Association of propofol and (S) ketamine for electroretinography in adult sheep M. Bacellar, H. D. Parchen, F. Montiani-Ferreira, A. T. Somma, I. R. Barros Filho and R. G. D.’OC Vilani Universidade Federal do Paraná, Curitiba, Paraná, Brazil Purpose: To investigate the use of the combination of (S) ketamine and propofol as anesthetic agents for electroretinography (ERG) in sheep. Methods: ERGs were performed in twelve sheep using the ISCEV protocol (HMsERG, RetVetCorp, Inc., Columbia, Missouri). Briefly, a 20 minute dark adaption time was used for before anesthetic induction scotopic ERGs and a 10 minutes light adaption time was used for photopic ERGs. Flash intensities used for scotopic ERG were 10, 3000 and 10 000 mcd.s/m2. For photopic ERG and 30 Hz-flicker the flash intensities used were 3000 and 10 000 mcd.s/m2. Sheep were premedicated intravenously with diazepam (0.5 mg/kg, Compaz, Cristália, São Paulo, Brazil) and morphine (0.5 mg/kg, Dimorf, Cristália, São Paulo, Brazil). In two groups of six animals each, anesthesia was induced with (S) ketamine (5 mg/kg, Ketamin-S(+), Cristália, São Paulo, Brazil) in the ketamine group (KG) or, with the combination of propofol (3 mg/kg, Propovan, Cristália, São Paulo, Brazil) and (S) ketamine (1 mg/kg) in the ketamine-propofol group (KPG). Half of the induction dose was used, and a quarter of the induction dose, were administered after 10 minutes to maintain the anesthetic level, in KG and KPG respectively. The following physiologic parameters were evaluated: rectal temperature, respiratory rate, cardiac rate, non-invasive arterial blood pressure, blood oxygen saturation. Corneal sensitivity was measured using a Couchet-Bounnet esthesiometer. Statistical analyses to compare ERG responses and all the physiologic parameters were performed using t-test and ANOVA, and the Exact Fisher test for esthesiometry. Results: All sheep of KPG and none of KG presented akinesia and centralization of the globe. The globes of KG animals needed stay sutures to perform the ERG. No significant differences in a- or b-wave amplitudes as well as implicit times were observed, comparing the two groups in any ERG test. No significant differences in corneal sensitivity, rectal temperature, cardiac rate, respiratory rate, SpO2 and mean arterial pressure were found. Significantly higher values were observed in diastolic and systolic arterial pressures immediately after the anesthetic induction in KPG compared with KG (P < 0.05). No further statistic differences were observed in any anesthetic parameters in both groups at any anesthetic moment. Conclusion: The association of the suggested doses of propofol and (S) ketamine promote akinesia and centralization of the globe, facilitating ERG, without important electroretinographic and physiologic alterations. ABSTRACT NO.: 084 Electroretinographic findings in sheep treated with the highest therapeutic dose of closante M. Bacellar, F. Montiani-Ferreira, H. D. Parchen, R. G. D.’OC Vilani, A. T. Somma, and I. R. Barros Filho Universidade Federal do Paraná, Curitiba, Paraná, Brazil Purpose: To investigate possible ERG changes in sheep that received the highest therapeutic dose of closantel. Methods: Twenty four 30 day old Sulffolk sheep were split into Control Group (CG, 12 animals) and Treatment Group (TG, 12 animals). The latter group received subcutaneously 15 mg/kg of closantel (Zantec, Biofarm, São Paulo, Brazil) immediately after the first ERG. For ERGs the sheep were premedicated with diazepam (0.5 mg/kg, Compaz, Cristália, São Paulo, Brazil) and morphine (0.5 mg/kg, Dimorf, Cristália, São Paulo, Brazil) IV, then anesthetized with propofol (3 mg/kg, Propovan, Cristália, São Paulo, Brazil) and (S) ketamine (1 mg/kg, Ketamin-S(+), Cristália, São Paulo, Brazil) IV. To maintain adequate anesthetic level, a quarter of the total induction dose was administered at 8 and 14 minutes after 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 402 a b s t r a c t s the induction. The ISCEV protocol (HMsERG, RetVetCorp, Inc., Missouri, USA) was used with a 20 minutes dark adaption for scotopic and a 10 minute light adaption for photopic ERG, respectively. Flash intensities for scotopic ERG were: 10, 3000 and 10 000 mcd.s/m2. For photopic ERG and 30 Hz-flicker the flash intensities used were: 3000 and 10 000 mcd.s/m2. This protocol was repeated at 7 and 30 days. Statistical analyses to compare ERG responses using t-test and ANOVA were performed. Results: No significant differences were found comparing the ERGs between two groups. CG, showed a significant increase (P < 0.05) of the mean scotopic b-wave implicit time from 0 to 30 days and an increase of the mean scotopic a-wave implicit time from 0 to 7 days, with the stimulus of 10 000 mcd s/m2. CG also showed a decrease in mean photopic b-wave implicit time (from 0 to 30 days), using a stimulus of 10 000 mcd s/m2 and a decrease in mean photopic a-wave implicit time from 0 to 30 days, using a stimulus of 3000 mcd s/m2. These differences observed in the CG were not observed in the TG. TG, however, showed a significant increase of the mean scotopic a-wave amplitude from 0 to 7 days after closantel administration, using a stimulus of 10 000 mcd s/m2 and a decrease of the mean scotopic and photopic a-wave amplitude (from 7 to 30 days) using the same flash intensity. TG also showed a decrease in mean photopic b-wave amplitude (from 7 to 30 days) within group. Conclusions: As observed by ERG responses closantel does not significantly affect retinal responses in sheep, when comparing with the control group. It does affect, however, normal ERG response development concerning wave amplitudes and implicit-times. ABSTRACT NO.: 085 Protective effects of BDNF and phenytoin against NMDA induced retinal damage S. Iwabe,* F. Gil-Carrasco,† A. A. Rodrı́guez-Reyes† and M. Lamas-Gregori* *Department of Pharmacobiology, Centro de Investigación y de Estudios Avanzados del IPN (Cinvestav), Mexico City, Mexico, †Glaucoma Service and Pathology Service from Asociación Para Evitar la Ceguera en México, Hospital Dr. Luis Sanchez Bulnes, IAP, Mexico City, Mexico. Purpose: To evaluate the neuroprotective effects of brain-derived neurotrophic factor (BDNF) and the anticonvulsant phenytoin against N-methyl-D-Aspartate (NMDA) induced retinal damage. Material and Methods: Forty rats (Long Evans) were injected intravitreally with NMDA (20 mM) to induce retinal excitotoxicity. Different groups of 10 animals each were injected with BDNF (0.5 lg) intravitreally and/or with phenytoin (100 mg/kg) intraperitoneally, one group did not receive any treatment. Right eye had different treatments while left eye had saline solution and was used as control. After 1 week animals were euthanized, eyes enucleated and subsequent morphometric analysis was carried out to evaluate the retinal cell damage. To elucidate BDNF and phenytoin neuroprotective effects, the retina were stained using Nissl staining (0.1% cresyl violet) to count retinal ganglion cells (RGC), and TUNEL technique was used to evaluate the apoptosis. Results: TUNEL technique showed major RGC death in the first 24 hours after NMDA administration. Statistical significance was observed between control eyes and eyes treated with BDNF or phenytoin separately, however no statistical difference was observed in the group treated with BDNF plus phenytoin (P > 0.05). Conclusion: In an acute environment of excitotoxicity induced with NMDA, the treatment using BDNF and phenytoin together provides better neuroprotection to retinal ganglion cells, when compared to control eyes. ABSTRACT NO.: 086 Diabetic retinopathy and cataracts in an aged raccoon with pancreatic islet amyloidosis C. M. Reilly,* B. C. Barr† and N. L. Anderson‡ *University of California, Davis Veterinary Medical Teaching Hospital, CA, USA, †California Animal Health and Food Safety Laboratory, Davis, CA, USA, ‡Lindsay Wildlife Museum, Walnut Creek, CA, USA Purpose: To report the clinical and pathologic findings in a raccoon with islet amyloidosis, diabetes mellitus (DM), and associated ocular lesions. Methods: This is a retrospective case report. The raccoon was submitted to the California Animal Health and Food Safety Laboratory for routine necropsy. Clinical history was reviewed prior to the necropsy, and a full necropsy was conducted. Tissues were routinely processed and key tissues were reviewed by two pathologists (CMR, BCB). PAS and Congo red stains were used where appropriate. Results: Pertinent history provided at necropsy included a 4 month history of cataracts and 6 years of variable hyperglycemia (218–359 mg/dL) without clinical signs. Prior hyperglycemia was attributed to alpha agonist sedation. Routine anesthetized exam after cataract development revealed blood and urine glucose levels of 208 mg/dL and 150 mg/dL, respectively, leading to the diagnosis of DM. Dietary modification corrected glucosuria, but was not well tolerated. He was euthanized due to progressive weight loss and terminal dyspnea. At necropsy, the pancreas was markedly enlarged, and there were complete, bilateral cataracts. Histologically, pancreatic islets were markedly widened by fibrillar eosinophilic material, which was confirmed to be amyloid with Congo red staining. Both eyes had hypermature cortical cataracts with liquefaction, and there was severe intraocular hemorrhage in one eye. Both retinas were completely detached, with multifocal supra- and sub retinal fibrovascular membranes. Retinal vessels were diffusely devoid of endothelial cells, with marked thickening of arteriole walls by homogenous, brightly eosinophilic, and PAS-positive material. Other ocular lesions included chronic uveitis, corneal neovascularization, and varying degrees of preiridal fibrovascular membrane formation. Conclusions: This is the first report of ocular manifestations of DM in raccoons to our knowledge. Raccoons are documented to undergo age-related islet amyloidosis, and there are isolated reports of DM in raccoons, but the two findings have not been correlated. The vascular and neovascular lesions in this case are similar to those described for diabetic retinopathy, which is common in humans, but rare in dogs with DM. The raccoon may be a model for diabetic retinopathy, but the apparently slow natural development of DM (>5 years) and diabetic retinopathy (>10 years) may warrant investigation of experimental induction. ABSTRACT NO.: 087 Goblet cell MUC5AC mRNA in canine nictitating membrane Y. Umeda,*,‡ S. Nakamura,† K. Fujiki,* H. Toshida,* A. Saito*,‡ and A. Murakami* *Department of Ophthalmology, Juntendo University School of Medicine, Tokyo, Japan, †Division of Biomedical Imaging Research, Biomedical Research Center, Juntendo University School of Medicine, Tokyo, Japan, ‡Triangle Animal Eye Clinic, Tokyo, Japan Purpose: Purpose of this study is to evaluate the distribution of the goblet cells and expression of MUC5AC mRNA in canine nictitating membrane. Methods: Distribution of the goblet cells in canine nictitating membrane was examined by histochemical analysis using frozen sections and impression cytology specimens in beagles. The detection of the MUC5AC mRNA was examined by reverse transcription polymerase chain reaction (RT-PCR). And distribution of the MUC5AC mRNA was also examined by in situ hybridization using digoxigenin (DIG)-labeled anti-sense and sense RNA probes. Results: In histochemical analysis, the canine nictitating membrane epithelium contains numerous PAS reaction positive goblet cells, particularly on the palpebral side. RT-PCR analysis detected that MUC5AC have been expressed in the canine nictitating membrane tissues. Distribution of MUC5AC mRNA on the canine nictitating membrane was much in the palpebral side as same as the histochemical examination. Conclusions: In the present study, we recognized numerous goblet cells in canine nictitating membrane epithelia, particularly the palpebral side, and the MUC5AC mRNAs have been expressed in canine nictitating membrane epithelia, corresponding to the goblet cells. ABSTRACT NO.: 088 Vitreoretinal surgery in Shih-Tzu with giant tear Y. Umeda, S. Wakaiki and A. Saito Triangle Animal Eye Clinic, Tokyo, Japan Purpose: To evaluate the success rate of the vitreoretinal surgery for giant tear associated with vitreous degeneration in Shih-Tzu. Methods: The 20G vitreoretinal surgery was performed for 14 Shih-Tzu with bilateral giant tear. The surgery was performed for one eye in every case. The eye performed the surgery was selected because of the latter onset by ophthalmic examination. Results: The repositioning of retina anatomically was found in 11 cases (78.6%) and functional recovery was recognized in six cases (42.9%). In two cases (14.3%), retinal evaluations after the surgery could not be performed because of the posterior synechia. And In one case (7.1%), the retina anatomical repositioning could not be performed because of the retinal synechia. The span of the functional recovery of the retina after surgery was 12–61 days (Average ± SD: 43 ± 24 days). Conclusions: We have been able to repositioned the retina anatomically in 11 cases, but recognized recovery of the retina functionally only in six cases. So it was thought that more consideration was required for preoperative examination. And as for two cases that a retina was not able to examine after surgery, the surgical invasion for the eye may have been strong. So it was thought that it may be useful to introduce less invasive surgeries like 23 G and 25 G vitreoretinal surgery systems. ABSTRACT NO.: 089 Evaluation of efficacy and biocompatibility of indirect intraocular pressure monitoring using a telemetric scleral sensor C. Robinson, D. Wilkie, A. Metzler, S. Weisbrode and R. Spatola College of Veterinary Medicine, The Ohio State University, OH, USA Purpose: To evaluate efficacy, biocompatibility and stability of a surgically implanted telemetric scleral sensor (TSS). To obtain an intraocular pressure (IOP) measurement without the need for topical anesthesia, patient compliance or corneal contact as is necessary with conventional applanation or rebound tonometry. Methods: A TSS was surgically implanted subconjunctivally and sutured to the sclera of the right eye of 10 juvenile female NZW rabbits. IOP was monitored using the TSS and a Tono-Vet twice daily for two weeks, then twice weekly until sacrifice. Ophthalmic examination was performed weekly and the eyes were graded according to the modified Hackett-MacDonald scheme. Two rabbits were sacrificed at 2, 4, 8, 12 and 16 weeks. Manometric IOP values were also recorded in 5 mmHg intervals up to 70 mmHg and compared to TSS. Eyes were submitted for histopathology. Results: Fibrosis and inflammation between the sclera and implant were graded on a scale of 0–4 for extent and severity, respectively. The grade of fibrosis and inflammatory response ranged from 1–4 (mean 2.5) and 0–2.3 (mean 1.2), respectively and was not associated with time. The predominant cell types recorded were the granulocyte and lymphocyte. Scleral thinning was observed at the site of TSS contact. Initial readings showed good correlation between the TSS and Tono-Vet, but premature dislocation of the TSS in six rabbits limited collection of manometric data. Conclusions: Although initial readings showed good correlation between the TSS and Tono-Vet, the overall size and design of the TSS contributed to fibrosis and premature dislocation of the sensor. Funding provided by Mesotec GmbH. ABSTRACT NO.: 090 Pharmacokinetics of famciclovir and penciclovir following single-dose oral administration of famciclovir to cats S.M. Thomasy,* T. Whittem,† S. D. Stanley* and D. J. Maggs* *School of Veterinary Medicine, University of California-Davis, CA, USA, †Faculty of Veterinary Science, University of Melbourne VIC, Australia. Purpose: To further investigate pharmacokinetics of famciclovir and its active metabolite penciclovir following oral administration of famciclovir to cats. Methods: Six cats (mean body weight 4.2 kg and age 1.3 years) each received a single oral dose of famciclovir at 40 mg/kg followed after a 14-day washout period by 90 mg/kg. At fixed time points for 24 h after famciclovir administration, plasma concentrations were determined for both famciclovir and penciclovir, using liquid chromatography/mass spectrometry, and concentration-time data assessed using noncompartmental analysis. A two-sample t test was used to determine whether dose had a significant (P < 0.05) effect on pharmacokinetic variables. Results: (mean ± SD). Following administration of 40 or 90 mg/kg famciclovir, maximum plasma famciclovir concentration (Cmax; 2.70 ± 2.23 or 2.98 ± 1.30 lg/ml) occurred at 0.8 ± 0.3 or 1.1 ± 0.5 h, and plasma penciclovir Cmax (1.34 ± 0.33 or 1.27 ± 0.33 lg/ml) occurred at 2.8 ± 1.8 or 3.0 ± 1.1 h, respectively. Penciclovir elimination half-life (t1/2(el)) was 4.2 ± 0.6 or 4.8 ± 1.4 h, respectively. Relative bioavailability of famciclovir following dose escalation from 40 to 90 mg/kg was 161% ± 124% (range 34–333%). No significant differences were detected in plasma Cmax or time to Cmax (Tmax) for either drug, or for penciclovir t1/2(el) for cats receiving 40 or 90 mg/kg of famciclovir. Conclusions: These data suggest that 40 mg/kg famciclovir produces similar Cmax, Tmax, and therefore potentially similar antiviral efficacy to that produced by 90 mg/kg famciclovir. Supported by the ACVO VAF. 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 abstracts 403 ABSTRACT NO.: 091 Substratum topography modulates TGFß-induced aSMA and smad7 exoressuin un rabbut cirneak strinak cells K. E. Myrna,* P. Russell,† S. J. Liliensiek,* J. V. Jester,‡ P. F. Nealey,§ D. Brown§ and C. J. Murphy‡ *School of Veterinary Medicine, University of Wisconsin, Madison, WI, USA, †School of Veterinary Medicine, University of California-Davis, Davis, CA, USA, ‡The Gavin Herbert Eye Institute, University of California-Irvine, Irvine, CA, USA, §College of Engineering, University of Wisconsin, Madison, WI, USA Purpose: TGFb is known to be important in regulating corneal fibroblast differentiation to the myofibroblast phenotype and plays an important role in normal and dysregulated wound healing. Myofibroblasts are generally located at the normal and wound tissue interface suggesting that the tissue provides cues that may modify TGFb signaling. The purpose of this study was to determine the effect of topographic cues on TGFb-induced myofibroblast differentiation of rabbit corneal stromal cells. Methods: Rabbit corneal fibroblasts were plated on nanopatterned surfaces having topographic features of varying pitch (pitch = groove width + ridge width). Cells were cultured with TGFb at concentrations of 0.0 ng/ml, 0.1 ng/ml, 1 ng/ml or 10 ng/ml. 72 hours later RNA was collected from the surfaces and analyzed for aSMA and SMAD7 by quantitative real time PCR. Results: Cells grown on a planar surface expressed negligible levels of aSMA at 0.0 and 0.1 ng/ml TGFb. A 3 fold increase was observed at 1 ng/ ml and a 23 fold increase at 10 ng/ml. Cells grown on all topographically patterned surfaces demonstrated significantly reduced levels of aSMA (P < 0.002) compared to the flat surface when exposed to at least 1 ng/ml of TGFb. Cells on the 1400 nm pitch surface expressed the lowest levels of aSMA, expressing 30% less than those on planar surfaces at 1 ng/ml TGFb and 90% less aSMA at 10 ng/ml TGFb. Cells grown on planar surfaces at all concentrations of TGFb showed equal and low levels of SMAD7. SMAD7 expression was significantly greater on all patterned surfaces with TGFb concentrations of 1 ng/ml and 10 ng/ml (P <0.002). There was a seven fold increase in expression at 400 nm and a four fold increase at 1400 nm at a TGFb concentration of 10 ng/ml. Conclusions: These data suggest that nanoscale topographic features modulate TGFb-induced myofibroblast differentiation and the expression of aSMA perhaps via a mechanism of SMAD7 inhibition. We propose that in the wound environment the native nanotopographic cues likely assist in stabilizing the keratocyte/fibroblast phenotype while pathologic alterations of the topographic environment may be permissive for myofibroblast differentiation and the development of fibrosis and corneal haze. Funded by NIH Grant 5R01EY012253-07. ABSTRACT NO.: 092 Characterization of ‘‘sea lion keratitis’’ in captive Zalophus californianus C. M. H. Colitz,* M. S. Renner,†,‡,†† C. A. Manire,‡ T. Reidarson,§ T. Schmitt,§ L. Croft,– C. Dold,– L. Dalton,** S. Osborn** and K. Terrell†† *Animal Eye Specialty Clinic, West Palm Beach, FL; Aquatic Animal Eye Care, Jupiter, FL; North Carolina State University and The Ohio State University Colleges of Veterinary Medicine, †Theater of the Sea, Islamorada, FL, USA, ‡Dolphin Cay Atlantis Paradise Island, Nassau Bahamas, §Seaworld CA San Diego, CA, USA, –Seaworld FL Orlando, FL, USA, **Seaworld Tx, San Antonio, TX, USA, ††Dolphin Encounters Blue Lagoon Island, Bahamas Purpose: To describe three consistent, progressive presentations of keratitis in Zalophus californianus. Methods: All animals underwent a complete ophthalmic examination and eyes were photographed. Results: One-hundred twelve eyes from 56 animals (42 males/14 females) were examined. Average age was fourteen years. Category one includes 19 animals with small, white and focal lesions at the dorsotemporal paraxial cornea, four had superficial ulcers. Four were unilateral and thirteen were bilateral. Category two includes seven animals with larger diffuse white corneal opacities that encompassed 5 to 20% of the dorsotemporal paraxial cornea, one was ulcerated. Six were unilateral, one was bilateral. Category three includes 11 animals with lesions encompassing over 20% of the cornea with stromal involvement. Most cases had documented bacterial and/or fungal infections while in stages 2 or 3. Three were unilateral, eight were bilateral. Conclusions: The initial cause of the stage one lesion is unknown. Possible etiologies include virus, exposure to UV/sunlight, and trauma. Viral testing has been negative, the consistent location makes trauma unlikely. Excessive UV exposure weakens and thins the corneal stroma and is likely part of the etiology. It appears that the recurrent focal initial lesion causes the cornea to become susceptible to secondary infections and this is likely exacerbated by excessive sun exposure. Further investigation into the primary cause as well as an imbalance of the local surface immune system will be pursued. ABSTRACT NO.: 093 Effect of central corneal thickness on intraocular pressure with the rebound and the applanation tonometers in dogs Y. W. Park, M. B. Jeong, T. H. Kim, S. E. Kim, J. S. Ahn, J. T. Ahn, S. A. Park, W. T. Kim and K. M. Seo Department of Veterinary Ophthalmology, College of Veterinary Medicine and BK21 Program for Veterinary Science, Seoul National University, Seoul, Korea Purpose: To evaluate the effect of central corneal thickness (CCT) on intraocular pressure (IOP) measurement with the rebound tonometer (TonoVet) and the applanation tonometer (TonoPen XL) in dogs. Methods: Both eyes of 60 clinically normal beagle dogs were included in the study. IOP was measured by TonoVet followed by TonoPen XL in half of them, and the other half were applied reverse order to reduce the examination order error. All CCT measurements by an ultrasonic pachymeter (DGH 55) were performed after IOP readings. All measurements of IOP and CCT were performed on the central cornea. Results: The mean IOP values measured by the TonoVet (16.9 ± 3.7 mmHg) were significantly higher than those of the TonoPen XL (11.6 ± 2.7 mmHg) (P < 0.001). The regression analysis showed that both IOP values were correlated (c2 = 0.4393, P < 0.001). Also, 95% values (114 eyes) were distributed within 95% limits of agreement in Bland-Altman analysis. The mean CCT was 549.7 ± 51.0 lm. There was a correlation between IOP values by two tonometers and CCT readings in regression analysis (TonoVet : P = 0.002, TonoPen XL: P = 0.035). The regression equation showed that every 100 lm increment in CCT results in 1 and 2 mmHg elevation of IOP values measured by the TonoPen XL and the TonoVet, respectively. Conclusions: It is considered that the IOP values are affected by the CCT in dogs. The IOP readings of the TonoVet were more affected than those of the TonoPen XL by the CCT. Therefore, the CCT is an important factor to be considered in interpretation of IOP values in dogs. ABSTRACT NO.: 094 Assessment of postoperative flare value following cataract surgery using a laser flare meter – influence on flare value by difference of IOL T. Yogo,* S. Kudo,† Y. Harada,* Y. Nezu,* Y. Hara* and M. Tagawa* *School of Veterinary Medicine, Nippon Veterinary and Life Science University, Department of Veterinary Surgery Tokyo Japan, †Kudo Small Animal Hospital, Tokyo, Japan Purpose: To investigate difference of flare value following phacoemulsification and implantation of a rigid IOL and a foldable IOL, based on flare values measured using a laser flare meter within 3 days after cataract surgery. Methods: Phacoemulsification and implantation of a rigid IOL (Meni-OneDV-20) was performed on 24 eyes in 17 dogs with cataracts and implantation of foldable IOL (Dioptrix PFI 14 SE) was performed on 13 eyes in 12 dogs with cataracts. We observed chronological changes in flare values using flare meter following the surgery. The flare value of the foldable IOL (Dioptrix PFI 14 SE) group was compared with rigid IOL group by two-way ANOVA analysis. Results: Flare values increased rapidly one day post-surgery but decreased markedly three days post-surgery. There was no statistically significant difference in the flare value between the rigid IOL and foldable IOL groups. Conclusions: The laser flare meter is a useful objective assessment tool of blood aqueous barrier breakdown with surgical damage and/or uveitis. Flare values increased rapidly one day post-surgery but decreased markedly three days post-surgery and remained low thereafter. As for the influence of the type of the intraocular lens on flare value, there was no significant difference between DV-20 and PFI 14 SE. Thus, it was suggested that the type of IOL implanted have little influence on flare value for a short term following cataract surgery. ABSTRACT NO.: 095 Visual examination with visual evoked potential using flash stimulus in dogs Y. Itoh, S. Maehara and Y. Izumisawa Rakuno Gakuen University, School of Veterinary Medicine, Department of Small Animal Clinical Sciences, Hokkaido, Japan Purpose: To evaluate visual examination with flush VEP (f-VEP) in dogs. Materials and Methods: Seven normal beagles (normal group) and four cases with visual impairment (disease group) were assessed. Dogs in disease group were diagnosed with cataract, optic nerve atrophy, chronic glaucoma and brain tumor adjacent in optic chiasm, respectively. VEP was recorded with plate-type electrodes positioned at inion, nasion and temporal region. The flash intensity was 3.0 cd/m2/sec, and 128 repetitions were averaged. In normal group, VEP was recorded before and after mydriasis. To evaluate the reproducible, VEP was recorded from same dogs 7 days later again. In disease group, VEP was recorded only with mydriasis. Results: In normal group, implicit time of N2 and P2 and amplitude of N2-P2 after midriasis were significantly shortened and increased compared with those before mydriasis, respectively. And there was significant difference in amplitude compared with first recording result. In disease group, VEP was obtained from cataract case. However, no VEP were recorded from cases of optic nerve atrophy, chronic glaucoma and brain tumor. Conclusions: Implicit time and amplitude are influenced by pupil size. Our results indicate that dogs should be in midriatic condition on f-VEP recording and implicit time is superior in the evaluation. Further, VEP could not be recorded from dogs with disorder regarded in post-retinal. Thus, we consider f-VEP is useful to objective their vision in clinical cases. This study is supported by Grant-in-Aid for Scientific Research from Japan Society for the Promotion of Science (No. 21780292). ABSTRACT NO.: 096 Effects of tafluprost 0.0015% compared with latanoprost 0.005% on the IOP and pupil size of normotensive dogs N. Takiyama, M. Sakai, H. Koie and M. Uechi Laboratory of Veterinary Internal Medicine, Department of Veterinary Medicine, College of Bioresource Sciences, Nihon University, Kameino, Fujisawa-shi, Kanagawa, Japan Purpose: To compare the effects of tafluprost 0.0015% and latanoprost 0.005% on the normotensive dogs. Methods: Fourteen Beagle dogs were randomized to two groups: tafluprost was used in one group and latanoprost in the other group. A single drop of tafluprost or latanoprost was instilled unilaterally. Artificial tear was instilled on the contra lateral eyes in both groups. IOP and pupil size (PS) were measured at 0, 0.5, 1, 2, 3, 4, 5, 6, 7 and 8 hours after administration. Results: Mean IOP and PS were significantly reduced in the eyes treated with tafluprost and latanoprost, when compared with contra lateral eyes (P < 0.05). There were no statistically significant difference between the mean IOP and PS of eyes treated with tafluprost and latanoprost at all time intervals. Conclusions: Tafluprost 0.0015% significantly reduces the IOP in normotensive Beagle dogs. The effects on IOP and PS obtained with tafluprost 0.0015% is equally to that obtained with latanoprost 0.005%. Commercial interests. 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 404 a b s t r a c t s ABSTRACT NO.: 097 Outcome of gene therapy is enhanced by CNTF-induced photoreceptor deconstruction in dogs with advanced stages of primary cone function loss A. M. Komáromy,* J. S. Rowlan,* J. J. Alexander,† V. A. Chiodo,‡ W. W. Hauswirth,†,‡ G. M. Acland§ and G. D. Aguirre* *Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA, †Department Molecular Genetics and Microbiology, University of Florida, Gainesville, FL, USA, ‡Department Ophthalmology, University of Florida, Gainesville, FL, USA, §Baker Institute, Cornell University, Ithaca, NY, USA Purpose: To study the effect of intravitreal CNTF administration on the therapeutic efficiency of rAAV-mediated gene replacement therapy in adult dogs with CNGB3 achromatopsia. Methods: Eight achromatopsia-affected dogs were injected unilaterally with 12 lg (30 lL) CNTF (kindly provided by Dr. Rong Wen, Bascom Palmer Eye Institute), while the fellow eyes were injected with 30 lL of PBS. After 7 days, seven dogs (age range: 14–39 months) were treated bilaterally with subretinal injections of rAAV5-PR2.1-hCNGB3. The injected volumes varied between 140 and 200 lL containing 7.96 · 1011–4.02 · 1013 particles per mL. Full-field ERGs were recorded before, as well as 1 and 5 weeks after the CNTF injections. Retinas were collected from three dogs for measurement of transgene expression and cone-specific gene expression levels by qRT-PCR. Results: Baseline ERGs confirmed normal rod-mediated function, but lack of cone-mediated responses. One week following intravitreal CNTF application, conventional full-field ERG responses were almost completely extinguished, but remained normal in the PBS injected eyes. Rod ERG responses recovered at 5 weeks post CNTF injection. Cone function was restored only when gene therapy was combined with intravitreal CNTF administration. High levels of hCNGB3 transgene expression were found in all rAAV injected eyes, regardless of success (intravitreal CNTF)/failure (intravitreal PBS) in restoring cone function. Clinically, no long-term adverse effects were observed after intravitreal CNTF injection. Conclusions: Intravitreal administration of CNTF leads to a fully reversible depression of outer retinal function (Parton-Corr et al., 2009), and dedifferentiation of photoreceptors (Beltran et al., 2007) in dogs, a process we now term photoreceptor deconstruction. We posit that the improved success rate of cone directed gene therapy in older mutant animals may result from treatment of cones that have dedifferentiated, and are able to reform the molecular components of the outer segment following redifferentiation. Supported by EY13132, 19304, 06855, 01583, 07132, 08571, 11123, 15398, 17549, FFB, MVRF, The ONCE Int’l. Prize. Commercial Relationship: P (WW Hauswirth, AGTC). ABSTRACT NO.: 098 Multiple congenital ocular anomalies (MCOA) syndrome in two silver coat colored ponies A. M. Komáromy and J. S. Rowlan Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA Purpose: Equine Multiple Congenital Ocular Anomalies (MCOA) or Anterior Segment Dysgenesis (ASD) syndrome consists of various numbers of ocular abnormalities predominantly affecting the anterior segment of the eye. It has been well described in the Rocky Mountain and Kentucky Mountain horses. While the disease-causing gene mutation has not yet been identified, the chromosome interval for the MCOA locus has been mapped to a 4.9-megabase interval on ECA6q (Andersson et al. 2008). This interval also contains the PMEL17 gene, which when mutated is responsible for Silver coat color (also called Silver dapple). While the Silver coat color shows an autosomal dominant inheritance, the alleles at the MCOA locus are codominant leading to either the Cyst or MCOA phenotype, based on either the heterozygous or homozygous genotype for the mutant allele. Here we provide the first formal report of the MCOA syndrome in ponies. Methods: Two unrelated, Silver coat colored ponies were examined. Both were rescue animals with limited history and no pedigree information. Case 1 was a ~23-year old Shetland Pony gelding that presented for cataract evaluation. Case 2 was a ~4-year old pony mare with no complaints regarding ocular abnormalities. In addition to the ophthalmic evaluation, blood was collected and genomic DNA extracted for MCOA genotyping using the PMEL17ex11 marker. Results: Multiple bilateral abnormalities were identified in Case 1, consistent with the MCOA phenotype, including cornea globosa, iris hypoplasia, and nuclear cataract. The animal was homozygous for the mutant MCOA allele. Case 2 presented with temporal ciliary body cysts only, which was suggestive of the Cyst phenotype and supported by the heterozygous MCOA genotype. Conclusions: To the best of our knowledge, this is the first formal report about MCOA syndrome in ponies. The results of both clinical evaluation and genotyping were in agreement with the previously published findings in the Rocky Mountain horses. Supported by NIH grants K12-EY15398 and R01-EY19304. ABSTRACT NO.: 099 Changes of aqueous flare values in four breeds of dog after phacoemulsification with IOL implantation Y. Fujii and S. Kudo Kudo Animal Hospital, Tokyo, Japan Purpose: Breed differences, especially American Cooker Spaniel (Cooker) and Toy Poodle (Poodle), were reported in post-operative inflammatory reactions on the ocular surgery. To examine these reactions, aqueous flare values were measured in 4 breeds of the dog, including Cocker and Poodle, after phacoemulsification with IOL implantation. Methods: Four breeds of the dog, 37 eyes of 30 American Cocker spaniel (age 3.0 ± 1.7), 41 eyes 31 Toy Poodle (age 4.8 ± 2.8), 26 eyes of 20 Cavalier King Charles Spaniel (Cavalier; age 4.8 ± 2.0), 22 eyes of 16 Shiba-Inu (Shiba; age 5.4 ± 2.9) were used in this study. Aqueous flare values were measured using the LASER FLARE METER FM-500 (Kowa, Tokyo, Japan) from the day 1 to the day 4 after phacoemulsification with IOL implantation between January 2007 and March 2009 at our practice. Statistical analysis was done using one-way analysis of variance. Results: Aqueous flare values (mean ± SD, as photon counts/msec) on the day 1 were 115.9 ± 68.2, 149.9 ± 169.1, 119.7 ± 160.7, 93.6 ± 70.9 in Cocker, Poodle, Cavalier, and Shiba, respectively. Those values on the day 2 and the day 3 were 56.0 ± 61.2, 96.7 ± 158.4, 95.5 ± 203.5, 32.9 ± 17.7, and were 50 ± 60.7, 61.7 ± 103.9, 38.3 ± 53.5, 14.8 ± 4.9, respectively. On the day 4, those values were 46.2 ± 39.0, 36.2 ± 18.1, 41.7 ± 48.2 in Cocker, Poodle, and Cavalier, respectively. These values were gradually reduced by the day 3 or 4 after ocular surgery in all four breeds. No difference in a decrease pattern of those values was observed in four breeds of the dog. Conclusions: From these results, high aqueous flare values observed on the day 1 is caused by the operative stress. Since no breed difference in a decrease pattern of aqueous flare value were observed, it is suggested that post-operative management and/or treatment is an important factor to control on post-operative inflammatory reactions. ABSTRACT NO.: 100 Feline conjunctival surface adenocarcinoma: ten cases C. Naranjo, C. S. Schobert and R. R. Dubielzig School of Veterinary Medicine, University of Wisconsin-Madison, WI, USA Purpose: To describe a novel tumor, conjunctival surface adenocarcinoma, in 10 cats. Methods: The Comparative Ocular Pathology Laboratory of Wisconsin (COPLOW) database was searched for cases diagnosed with feline conjunctival surface adenocarcinoma. Information regarding the breed, age, gender, affected eye and location of the tumor within the conjunctiva was recorded. The referring ophthalmologist was contacted for follow up information. Histopathology was reviewed and Periodic Acid Schiff (PAS) stain to detect mucopolysaccharides was performed on at least one sample from each animal. Results: Ten cats were diagnosed with conjunctival surface adenocarcinoma, including five Domestic Long Haired, two Persian, two Siamese or Siamese cross and one Domestic Short Haired. Mean age was 10.7 years (range 3 to 18 years). Gender distribution was homogeneous. The most common location included conjunctiva overlying the third eyelid, although multifocal distributions were common. The tumor was frequently contiguous with the surface epithelium of the conjunctiva, forming slender papilliferous projections. Tumor cells frequently infiltrated adjacent structures, including the lamina propria of the conjunctiva, cornea at the limbus, and the gland of the third eyelid. Neoplastic cells were organized in solid packets, with occasional acini noted. PAS-positive material was found within the cytoplasm of neoplastic cells in six of 10 cases. PAS-positive extracellular material within the lumen of acinar structures was found in 4 of 10 cases. Local recurrence was frequent, so aggressive surgery, including enucleation and exenteration, was necessary in some cases. Conclusion: Feline conjunctival surface adenocarcinoma must be considered as a differential diagnosis for masses involving the conjunctiva and third eyelid of the cat or any multifocal nodular proliferations in the conjunctiva. Radical surgery may be required as this tumor has potential for recurrence if incompletely excised. ABSTRACT NO.: 101 A retrospective evaluation of the diamond burr superficial keratectomy in the treatment of spontaneous chronic corneal epithelial defects in dogs from 2006 to 2008 G. H. Sila, R. J. Morreale, D. W. Lorimer, H. J. Davidson and C. D. Bliss Department of Ophthalmology, Michigan Veterinary Specialists, Southfield, MI, USA Purpose: To determine approximate healing times and complications in dogs with spontaneous chronic corneal epithelial defects (SCCEDs) treated with a diamond burr superficial keratectomy (DBSK), grid keratotomy with a #64 beaver blade under anesthesia (GKA), or a grid keratotomy with a 25G needle under topical anesthesia (GKT) at a private referral hospital. DBSK has been used recently in human medicine for treatment of refractory epithelial erosions with good results. Methods: Eyes with a diagnosis of SCCED from 2006 to 2008 were eligible for inclusion. Records from patients that underwent GKA, GKT, or DBSK, and had no history of concurrent corneal disease were reviewed. Healing times, doctor performing the procedure, complications, post-operative antibiotic choice, and the need for any additional procedures were recorded. Statistical analysis was performed using Chi square contingency tables. Results: A total of 195 eyes were eligible for inclusion in the study (100 DBSK, 52 GKT, and 43 GKA). DBSK eyes were significantly more likely to have healed at weeks 1 to 2 and 2 to 3 than the other methods (P < 0.05). Complications following DBSK were rare and the most significant complication was development of varying degrees of keratomalacia early in the development of the technique (3/100 eyes). All three eyes improved with medical management. Conclusions: DBSK is a promising new method for the treatment of SCCEDs and may allow faster healing times than more traditional methods. Further evaluation of the DBSK as a treatment for SCCEDs is indicated. ABSTRACT NO.: 102 Effects of fixed combination timolol 0.5%/travoprost 0.004% compared with travoprost 0.004% on the intraocular pressure in normal dogs P. Jitasombuti,* S. Asawapureekorn,† T. Ployngam,* N. Petchoo,* K. Boonloh,* T. Somphabutr* and P. Wipoosak* *Faculty of Veterinary Medicine, †Faculty of Medicine, Khon Kaen University, Muang District, Khon Kaen, Thailand Purpose: To compare the effects of fixed combination timolol 0.5%/travoprost 0.004% and travoprost 0.004% on the intraocular pressure (IOP) in normal dogs. Methods: Ten mixed breed dogs were divided randomly into two groups of 5 each. Each group received instillation of either fixed combination timolol 0.5%/travoprost 0.004% or travoprost 0.004% on the right eyes at 8 AM. The left eyes in both groups served as control, which were instilled with normal saline at the same time. IOP were measured at 8 AM, 10 AM, 2 PM, and 8 PM during 3 days of treatment. Results: Mean IOP was significantly reduced by day 1 and 2 in the eyes treated with timolol 0.5%/travoprost 0.004% and travoprost 0.004%, respectively (P < 0.05). By the end of the study protocol, mean IOP was reduced by 3.1 ± 0.3 mmHg in travoprost 0.004% group and 1.1 ± 0.3 mmHg in timolol 0.5%/travoprost 0.004% group. However, no significant difference between the mean IOP of the eyes treated with timolol 0.5%/travoprost 0.004% and travoprosted 0.004% was observed (P > 0.05). Conclusions: Travaprost 0.004% and fixed combination timolol 0.5%/travoprost 0.004% significantly reduce the IOP in normal dogs. The ocular hypotensive effect of timolol 0.5%/travoprost 0.004% and travoprost 0.004% is comparable. 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 abstracts 405 ABSTRACT NO.: 103 Incidence of juvenile cataract in 213 Labrador retrievers for guide dog in Japan A. Kubo,* T. Yogo,† K. Terakado,† Y. Suwa‡ and T. Furukawa§ *Veterinary Eye Care Service, Takahashi Animal Hospital, Japan, †Department of Veterinary Surgery, Nippon Veterinary and Life Science University, Japan, ‡Hokkaido Guide Dogs for the Blind Association, §Department of Comparative Animal Science, Kurashiki University of Science and the Arts, Japan Purpose: To determine the incidence of juvenile cataracts in Labrador retrievers training for guide dog to control the incidence of cataracts with breeding plan. Methods: A total of 213 Labrador retrievers (114 males and 99 females) training for guide dog were examined with slit lamp biomicroscope to detect cataract. Age, gender, the familial relation and the position of cataracts in the lens were determined. Results: This study showed that cataracts were observed in 17 eyes of 12 dogs (six males and six females), the incidence rate was 5.63% and the average age of the detection was 1.24 ± 0.24 year old. Cataracts were detected at the posterior parts of lens including posterior cortex, suture and capsule in 11 of 12 dogs with cataract. There were two groups of two dogs from each same litter, a group of four dogs from same father and another group of two dogs from same father, two groups of three dogs from each same mother and a group of two dogs from same mother in the dogs with cataract. Conclusions: In this study, the incidence rate of cataracts was lower than the spontaneous incidence rate reported previously. No gender predisposition was observed. Most of cataracts were detected at the posterior part of lens as previous report in Labrador retrievers. The familial relation with cataract formation was observed significantly and we considered the genetic factor was strongly related to the cataracts observed in this study. ABSTRACT NO.: 104 Evaluation of the photopic negative response and oscillatory potentials in New Zealand white rabbits with elevated intraocular pressure following intra-cameral injection of 1% methyl cellulose and vortex vein ligation P. P. Ko,* G. G. Gum,* V. K. Ackley* and R. A. P. de Carvalho† *Biological Test Center, †3T Ophthalmics, Irvine, California Purpose: The objective of this study was to evaluate the photopic negative electroretinogram (ERG) response and oscillatory potentials (OP) over time in two models of retinal ganglion cell loss due to increased intraocular pressure (IOP) in New Zealand White rabbits—following intra-cameral injection of 1% Methyl Cellulose and vortex vein ligation. Methods: Ten New Zealand White rabbits were randomly assigned to receive in the right eye an intracameral injection of 0.25 mL of 1% methyl cellulose after an equal volume of aqueous humor was collected from a separate intracameral port. In the second group, the right eye had three out of four vortex veins ligated. ERG, IOP, OCT, fundus photography, fluorescein angiography and ophthalmic observations were performed at baseline, and at multiple points throughout the study. All rabbits were scotopically adapted for 20 minutes prior to the ERG procedures. ERGs were recorded with a 20 msec stimulus of scotopic luminance (-6dB-blue filter, 0dB, +20dB-white flash) and photopic luminance (+5dB-low intensity with blue background & red stimuli, +12.5dB-high intensity with blue background & red stimuli, +20dB-white flash). The implicit times and amplitudes of PhNR and oscillatory potential (OP) were recorded. Statistical analysis was performed by using Student t-test and ANOVA with multiple comparison procedures (level of significance P < 0.05). Results: In eyes with vortex vein ligation, the amplitudes of ERG PhNR and OP had both declined progressively over time compared with the baseline. By Day 9, the mean PhNR amplitudes at +5dB decreased by an average of 50% (P = 0.0210) and by 30% at +12.5dB (P = 0.1625) after vortex veins ligation. In the eyes after intracameral injection, the mean PhNR amplitudes at +5dB decreased by an average of 38% (P = 0.1434). The ERG data demonstrated that the decreased amplitudes and prolonged implicit times of OP are more sensitive and specific than the declining PhNR amplitudes. Conclusion: The decreased amplitudes of PhNR and OP indicate the damage at the ganglion cell level & inner retinal plexiform layer respectively is caused by the slow increased IOP. The amplitudes and implicit times of OP are sensitive and specific to monitor the possible early damage of IOP changes. ABSTRACT NO.: 105 Complete reconstruction of the superior eyelid in a dog utilizing a superficial temporal axial pattern flap and nictitans-based conjunctival pedicle graft B. J. Stanley, J. T. Bartoe and K. E. Pierce College of Veterinary Medicine, Michigan State University, East Lansing, MI, USA Purpose: Reconstruction of large, full-thickness defects of the superior eyelid can be challenging, as maintenance of lid function is critical for ocular health. This case report describes a novel, two-stage reconstruction of the skin, margin, and conjunctiva of the superior eyelid. Methods: An 11-year-old dog presented with a grade II mast cell tumor involving the left superior eyelid, with no evidence of metastatic disease. Two-centimeter wide excision was performed. The first stage reconstruction involved the development of two flaps, firstly, a superficial temporal axial pattern flap to restore tension-free cutaneous palpebral tissue, and secondly, a conjunctival pedicle flap based at the nictitans margin, and reflected from the palpebral surface of the third eyelid to provide an inner conjunctival lining to the neo-palpebra. A temporary tarsorrhaphy was placed during healing. In the second surgical stage three weeks later, the base of the nictitans flap was transected. The pigmented, cartilaginous leading edge of the nictitans was reflected over and sutured to the distal margin of the neo-palpebra, creating a well-defined superior palpebral margin. Post-operatively, the eye was medicated with a topical artificial tear gel and neomycin/polymixin/bacitracin ointment. Results: Histopathology found no evidence of neoplastic cells at any margins. The dog healed without complication, and with excellent cosmesis and corneal health. A partial functional blink emanating from the lower eyelid was observed, although continued daily artificial lubrication of the cornea was recommended. Conclusions: This two-stage technique allowed wide, curative excision of the neoplasia with preservation of the globe and excellent cosmetic outcome. ABSTRACT NO.: 106 Use of rebound tonometry as a diagnostic tool to diagnose glaucoma in the captive California sea lion J. C. Mejia-Fava,* L. Ballweber,† C. M. H. Colitz,‡ C. C. ClemonsChevis,§ L. Croft,– L. Dalton,** C. Dold,– S. Gearhart,– E. Hoffman,†† S. Osborn,** S. W. Jack,‡‡ M. Renner,§§ T. Schmitt,–– M. Rodriguez,§§ T. Romano*** and A. Tuttle*** *Grand Cayman Boatswain’s Beach Turtle Farm, †Colorado State University, Fort Collins, CO, USA, ‡Aquatic Animal Eye Care, Animal Eye Specialty Clinic, The Ohio State University College of Veterinary Medicine, Columbus, OH, USA, §Institute for Marine Mammal Studies, Gulfport, MS, USA, –Seaworld Florida, Orlando, FL, USA, **SeaWorld Texas, San Antonio, TX, USA, ††CPT, Veterinary Corps, US Army. U.S. Navy Marine Mammal Program SPAWAR Systems Center, San Diego, CA, US, ‡‡Mississippi State University College of Veterinary Medicine, Mississippi State, MS, USA, §§Miami Seaquarium, Miami, FL, USA, ––Seaworld California, San Diego, CA, USA, ***Mystic Aquarium and Institute for Exploration, Mystic, CT, USA Purpose: One of the most common medical problems seen in the California sea lion (Zalophus californianus, CSL) is ocular disease. Glaucoma is a disease that has not been evaluated extensively in the CSL. Observing clinical signs and measuring intraocular pressures (IOP) is critical for early diagnosis. The objective of this project is to measure IOP in clinically normal captive CSLs without ocular pathology to establish a normal range. Methods: The Tono-Vet (Web ster Veterinary) was selected to be used in the study. The Tono-Vet uses non-invasive, rebound methods to estimate IOP. In order to record an accurate IOP, six measurements were taken and averaged resulting with the mean value. A complete ophthalmic examination has been performed on all CSLs by a veterinary ophthalmologist. Results: Currently, there are twenty sea lions in the study with no clinical ocular pathology. Overall mean in 39 healthy eyes was 32.8 mmHg with a SD± = 3.2 at a 95% CI of 26.4 to 39.1. Conclusion: We have established a normal baseline range for IOP values in captive CSLs without ocular pathology. This range is higher than the generally accepted range using other tonometers (e.g. Tono-Pen Vet). This is likely due to the increased thickness of the pinniped cornea as well as the different mechanism of the instrument itself. This range will provide a comparative measurement when evaluating a diseased eye. By measuring the IOP regularly in juvenile CSLs, earlier management of glaucoma and its underlying cause can be implemented prior to loss of vision. ABSTRACT NO.: 107 Morgan pocket technic plus ciclosporine 0.2% ointment in dogs with prolapsed third eyelid gland P. D. Galera, R. V. R. Peixoto and M. S. A. Falcão College of Veterinary Medicine, University of Brasilia, Brazil Purpose: Prolapsed third eyelid gland can occur normally with keratoconjunctivitis sicca. This work‘s purpose is to evaluate the tear Schirmer test (STT-1) of dogs (19 eyes) with prolapsed third eyelid gland that were submitted to Morgan pocket technic and received ciclosporine 0.2% ointment. Methods: Fifteen dogs (19 eyes) with prolapse of the third eyelid gland were evaluated, and submitted to STT-1 They underwent the Morgan pocket surgery, and were pre (7 days) and post medicated (15 days) with ciclosporine 0.2%, twice a day, for 21 days. Results: In the first examination before the surgery, 67% of the eyes had other ophthalmic alterations aside from the prolapse of the third eyelid gland. The use of the medication before surgery reduced the inflammation. In the post operatory period, 70% of the eyes had normal clinical aspect and normal STT values. The mean of STT was 214.68 and 20.84 mm/min before and after surgery, respectively. In 89.5% of the cases the association of ciclosporine 0.2% and surgery showed good results. Conclusions: The Morgan pocket technique plus ciclosporine 0.2% improved the STT values and restored the clinical aspects of the eyes. Supported by FINATEC; FAPDF. ABSTRACT NO.: 109 Protein measurement of aqueous humor of clinically healthy dogs through unidimensional electrophoresis gel (SDS-PAGE) with color in silver M. A. S. Falcão,* P. D. Galera,* R. V. R. Peixoto,* T. C. Ferreira† and E. G. Campos† *College of Veterinary Medicine, University of Brasilia, Brazil, †Cellular Biology Laboratory, University of Brasilia, Brazil Purpose: To describe the standardization of the technique for measuring the protein profile of the aqueous humor of clinically healthy dogs, through the differentiation of proteins by molecular weight by gel-dimensional electrophoresis (SDS-PAGE) with Color in Silver and densitometry. Methods: A total of 23 clinically healthy adult mixed breed dogs were selected without ophthalmic problem, eight females and 15 males. The animals were anesthetized and subjected to paracentesis of anterior chamber and a sample of 0.3 mL of aqueous humor of each eye was obtained. The material was centrifuged and put in electrophoresis gel and stained with silver nitrate. Results: The molecular weight of proteins present in the aqueous humor averaged between 120 and 25 kDa, according to the molecular mass marker (190–15 kDa). One of the protein bands, present in all samples, has a molecular weight equivalent to the molecular weight of BSA, inferring that it is the canine albumin. This protein presented the higher molecular weight between the proteins. Conclusions: We conclude that the technique described above is valid for the analysis of proteins of the aqueous humor of healthy dogs. Supported by FINATEC; FAPDF. 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 406 a b s t r a c t s ABSTRACT NO.: 110 Quantitative analysis of aqueous humor protein of dogs with cancer P. D. Galera,* C. M. Nasser,* M. A. S. Falcão,* M. S. T. Rocha,* T. C. Ferreira† and E. G. Campos† *College of Veterinary Medicine, University of Brasilia, Brazil, †Cellular Biology Laboratory, University of Brasilia, Brazil Purpose: The aim was to quantify the aqueous humor proteins in dogs affected by cancer using the Bradford’s colorimetric test and to compare it to the quantity found in healthy dogs. Methods: Six dogs with cancer (one sarcoma histiocytic; two osteosarcoma; one squamous cells carcinoma, one hemangiosarcoma, one mast cell) and three healthy dogs (control group) had their aqueous humor collected (0.3 mL) by anterior chamber paracentesis. The amount of aqueous humor protein was measured by the Bradford method. Blood was also drawn for total protein blood quantification. Results: A reduction of the protein levels was found in the left eye of sick dogs (8.45 mg/dl) compared statistically to the amount of the protein levels found in the left eye (20.47 mg/dl) of the control group. A significantly statistic result was also found between the right eye (17.46 mg/dl) and the left eye (8.45 mg/dl) of the sick dogs, showing a reduction of the left eye’s protein levels, which was not observed in healthy animals. The values of aqueous humor protein of dogs with cancer did not correspond to the 0.5% of plasma protein values found in healthy dogs. Conclusion: The hemato-aqueous barrier was intact to systemic alterations in animals with cancer. Other researches should be done to elucidate this initial correlation. Supported by FINATEC; FAPDF. ABSTRACT NO.: 111 Neovascular proliferative vitreoretinopathy reminiscent of retinopathy of prematurity: a morphologic and immunohistochemical study in cats A. Hoffman,* K. Tawansy,† E. J. Ehrhart,‡ J. Charles,‡ M. Zarfoss,§ C. Schobert– and R. R. Dubielzig– *Eye Care for Animals, Pasadena, CA, USA, †Children’s Retina Institute, Los Angeles, CA, USA, ‡Colorado State University, Fort Collins, CO, USA, §University of Illinois, Urbana-Champaign, IL, USA, –University of Wisconsin-Madison, WI, USA Purpose: This study aims to report clinical features, histopathologic and immunohistochemical results in a population of young cats with neovascular proliferative vitreoretinopathy. Methods: Cases diagnosed as neovascular vitreoretinopathy were selected from a total of 6158 feline submissions from COPLOW. Histopathology submissions were reviewed to obtain information on age, breed, gender, and clinical features. Routine H & E slides were reviewed to describe morphologic characteristics. Immunohistochemistry included GFAP, Collagen IV, laminin, SMA, CD68 and FVIIIRA and was performed on globes available for further sectioning. Results: Most cats presented at <1 year of age, with 8/14, <6 months old. Representative breeds were Persian (2), domestic medium hair (2) and domestic short hair (10). Among these breeds, eight were female and six were male. The most common clinical presentation was glaucoma. Histopathology: Retinal detachments were noted in 11/14 eyes. The retina was entirely avascular in one case, and 80–90% avascular in the remaining 13 cases. The course of retinal vessels traveled axially within the vitreous towards the lens in all cases. The nature of the vitreous tended to be very liquid. Fibrovascular membranes were found in all 14 cases. Anterior uveal inflammation was noted in 12/14 cases. Immunohistochemical findings: GFAP: showed excessive and atypical gliosis in the retina beyond what would be expected from just retinal detachment. Collagen IV and laminin: showed + staining for basement membrane on the inner border of the retina and continuous with the inner limiting membrane of the retina. SMA: showed myofibroblastic cells in connective tissue and supported that the retinal detachment resulted from traction. CD68: the presence of histiocytic cells were variable within vessels and membranes but did not appear to correlate with the proliferative response. FVIIIRA: confirmed that vascular endothelial cells are responsible for the vasoproliferative reaction. Conclusions: Feline retinopathy of prematurity is a vasoproliferative disorder with retinal detachment, avascular retina, and excessive retinal gliosis that should be considered a cause of feline pediatric uveitis or glaucoma. ABSTRACT NO.: 113 Equine amniotic membrane transplantation for canine and feline corneal diseases K. Kim,* J. S. Sapienza* and D. E. Brooks† *Long Island Veterinary Specialists, Plainview, NY, USA, †College of Veterinary Medicine University of Florida, Gainesville, FL, USA Purpose: To report the use of equine amniotic membrane transplantation in clinical cases of canine and feline ulcerative keratitis. Methods: Six dogs and one cat (n = 7 eyes) with ulcerative corneal diseases were treated with equine amniotic membrane transplantation (AMT, [University of Florida]) following poor response to medical therapy. Medical records were retrospectively reviewed to evaluate pre-operative clinical courses, surgical methods, and postoperative outcomes. Results: Three breeds of dogs (four Shih tzus, one Miniature pinscher, one Bichon frise) and one domestic longhair cat were treated with AMT for unilateral progressive keratitis. Two spayed females, two intact females, two neutered males and one intact male were treated. Two dogs received single layer AMT, and four dogs received double layer AMT. The cat had a corneal perforation and received an underlying Biosist graft in addition to a single layer AMT. Post-operative follow-up ranged from two weeks to two months. Graft dehiscence of the second overlying AMT was always observed. One dog developed bullous keratopathy that resolved with medical therapy. At the last reevaluation, all seven patients demonstrated progressive improvement with no recurrence or persistence of the underlying corneal disease. Conclusions: In cases of canine and feline deep ulcerative keratitis or keratomalacia that are refractory to medical therapy, AMT may be considered as an adjunct or alternative surgical therapy in order to optimize corneal healing and to minimize corneal scarring. ABSTRACT NO.: 114 Determination of progenitor cells during canine retinogenesis M. C. Ávila,* G. A. Garcı́a† and N. A. Moreno* *Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, †Facultad de Medicina Veterinaria y Zootecnia, UNAM, Mexico Purpose: To determine the expression of specific markers for stem, ganglion and proliferation cells into canine fetuses retinas. Future cell-replacement therapy with progenitor cells represents an alternative treatment for canine glaucomatous neurodegeneration. Methods: Eyes from canine fetuses (55 days of gestation) were enucleated and processed for immunohistochemical analysis. Expression of specific markers of neural stem cells (Pax6, c-kit and nestin), ganglion cells (BDNF) and cell proliferation (H3) was performed. Results: Expression of Pax6, nestin, c-kit and BDNF was localized mainly in the inner neuroblastic layer (INL), while cells in the outer neuroblastic layer (ONL), adjacent to the pigmented epithelium, expressed H3. Colocalization of Pax6/nestin, c-kit/Pax6 and BDNF/nestin was observed in the INL, while nestin/H3 was found in the ONL. Colocalization of c-kit/Pax6/H3 and nestin/Pax6/c-kit was found in the INL. Conclusions: During retinogenesis (55 days) the cells in the INL have molecular characteristics of stem cells, moreover some cells are already differentiated into ganglion cells. The ONL begins cell proliferation that will differentiate in photoreceptor cells. In fetuses of 55 days the cells of the INL showed specific markers of stem cells (Pax6, nestin and c-kit), while some ganglion cells have affinity to BDNF. Also cells in the ONL showed characteristics of cell proliferation, which subsequently initiate their differentiation into photoreceptors. Supported by DGAPA–IN218908 (UNAM). ABSTRACT NO.: 115 Exclusion analysis reduces the list of candidate genes for the ocular melanosis causative mutation in cairn terriers J. T. Bartoe,* P. A. Winkler,* C. Quinones,* P. J. Venta† and S. M. Petersen-Jones* *Department Small Animal Clinical Sciences †Department Microbiology and Molecular Genetics, College of Veterinary Medicine, Michigan State University, East Lansing, MI, USA Purpose: To use genetic markers to screen for linkage of candidate genes to the Ocular Melanosis (OM) locus in Cairn terriers. Methods: Eleven genes: ASIP, COMT, DCT, GPNMB, GSK3B, LYST, MC1R, MITF, SILV, TYR, and TYRP1 were identified as candidate genes potentially encoding OM causative mutations. Conserved polymorphic genetic markers consisting of single nucleotide polymorphisms (SNP), insertion/deletion (In/Del) or microsatellite copy variants (MS) were identified in or within 0.5 megabases of these 11 genes. PCR primer pairs were designed to amplify two unique markers per gene. SNP allelic variation was detected by evaluating the digestion pattern of PCR amplicons containing naturally occurring or primer-engineered restriction endonuclease sites. In/Del and MS alleles were typically easily discriminated on agarose gels. MS of indeterminate gel separation were sent for high-resolution genotyping. Previous pedigree analysis has demonstrated a dominant inheritance pattern for OM. Therefore, 5–10 unaffected Cairn terrier DNA samples were screened for marker allelic variation within the breed. Once polymorphic markers were found, 10–20 OM affected samples were evaluated for a single shared marker allele, assuming linkage between the marker and the OM locus. Results: No group of OM affected dogs was found to have a single shared marker allele. Conclusion: Exclusion analysis suggests ASIP, COMT, DCT, GPNMB, GSK3B, LYST, MC1R, MITF, SILV, TYR, and TYRP1 are unlikely to contain mutations resulting in OM in Cairn terriers. Supported by AKC-Canine Health Foundation, Cairn Terrier Club of America, MSU-CVM Pure-bred Dog Endowment Fund, Glassen Foundation. ABSTRACT NO.: 116 Distichiasis causing recurrent corneal ulceration in two Friesian horses K. L. Wotman and M. E. Utter New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, PA, USA Purpose: Two cases of distichiasis with subsequent recurrent corneal ulceration are described in Friesian horses. Method: Retrospective case review. Results: Case 1 was a 9-year-old Friesian stallion with distichiae on the superior and inferior eyelids of both eyes corresponding to areas of recurrent corneal ulceration. Case 2 was a 5-year-old Friesian gelding with a single distichia on the inferior lid associated with recurrent superficial corneal ulceration in the right eye. Case 1 underwent cryotherapy and epilation twice under standing sedation without resolution of distichiasis. A split lid procedure was then performed on the inferior left lid, which was the most severely affected. Cryotherapy was repeated 4 months post-operatively due to recurrence. Cryotherapy was associated with skin depigmentation and repigmentation several months following the procedure. No recurrence has been reported up to 18 months following the third cryotherapy procedure. Case 2 was treated with epilation. Familial relationship between the two horses has not been established. Conclusions: Distichiasis should be included as a differential diagnosis when presented with a horse experiencing recurrent superficial corneal ulcerations. There has been no established breed association for Friesian horses and distichiasis but both horses in this report were Friesians imported from Friesland, the Netherlands. ABSTRACT NO.: 117 Localization of interleukin 11 in the canine eye T. R. Richards, N. C. Whelan, C. Pinard, J. Mortlock and J. LaMarre Ontario Veterinary College, University of Guelph, Guelph, ON, Canada Purpose: Interleukin 11 (IL-11) is a pleiotropic cytokine with anti-inflammatory, cytoprotective, epitheliotrophic and immune modulating properties. The purpose of this study is to characterize the expression pattern of IL-11 in normal canine eyes. Methods: Normal canine eyes were collected from eight dogs. The distribution of IL-11 expression in the different ocular layers was evaluated by immunofluorescence. Expression levels were quantified (based on fluorescence intensity) using pixel density analysis. IL-11 expression was also assessed in primary cultures of corneal endothelium and fibroblasts using quantitative RT-PCR in untreated cells 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 abstracts 407 and cells treated with TGF-b, a known inducer of IL-11 expression. Results: Cytoplasmic expression of IL-11 was observed in the cornea and iridocorneal angle. Variable levels of IL-11 expression were noted in the iris and retina, and no expression was consistently present in the choroid, ciliary body or sclera. A low level of IL-11 mRNA was constitutively expressed in corneal fibroblast and endothelial cells. TGF-b exposure caused a >100-fold increase in IL-11 expression in both cell lines 12–24 hours after treatment. Conclusion: This is the first description of IL-11 expression in the canine eye. It is primarily expressed in the cornea and iridocorneal angle, and expression in the cornea is increased in the presence of TGF-b, a cytokine important in ocular inflammation and disease. Funded by the OVC Pet Trust. ABSTRACT NO.: 118 Expression of matrix metalloproteinases in feline eyes with uveitis and lens luxation J. J. Broadwater,* C. M. H. Colitz,*,§ R. R. Dubielzig,† S. M. Carastro* and H. L. Chandler‡,§ *Animal Eye Specialty Clinics of South Florida, Miami, FL, USA, †University of Wisconsin, School of Veterinary Medicine, Madison, WI, USA, ‡The Ohio State University, College of Optometry, §College of Veterinary Medicine, Columbus, OH, USA Purpose: To determine the expression of matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9) in normal feline eyes (N), feline eyes with uveitis (U), and feline eyes with uveitis and lens luxation (ULL). Methods: Zymography, followed by densitometry, was performed on aqueous humor samples from 4 N, 3 U, and 3 ULL to determine expression of MMP-2 and MMP-9. Immunohistochemistry was performed on 10 N, 10 U, and 10 ULL to evaluate the presence of MMP-2 and MMP-9 in the ciliary body and iris. Results: MMP-2 and MMP-9 were present in aqueous humor from both normal and diseased eyes. U samples had an average of 1.44 fold increase in MMP-2 expression and an average of 1.25 fold increase in MMP-9 expression compared to N. ULL samples had an average 1.60 fold increase in MMP-2 expression and an average of 1.42 fold increase in MMP-9 expression compared to N. Immunohistochemistry revealed subjectively increased staining for both MMP-2 and MMP-9 in ULL compared to both N and U samples. Conclusions: Increased levels of MMP-2 and MMP-9 were found in aqueous humor samples of cats with uveitis and cats with uveitis and lens luxation as compared to normal samples. Increased expression of MMP-2 and MMP-9 was also found in the iris and ciliary body of cats with uveitis and lens luxation, as determined by immunohistochemistry. MMP’s may be involved with the degradation of ciliary zonules and contribute to the development of lens luxation in cats with uveitis. ABSTRACT NO.: 119 A retrospective review of the histological and clinical characteristics of canine ocular gliovascular syndrome (cogs): a syndrome of intravitreal glial aggregates, neovascular membrane proliferation, intraocular hemorrhage, and neovascular glaucoma in the dog A. N. Treadwell,* C. Naranjo,† M. K. Zarfoss,‡ T. Blocker* and R. R. Dubielzig† *Eye Care for Animals, †University of Wisconsin College of Veterinary Medicine, Madison, WI, USA, ‡University of Illinois College of Veterinary Medicine, Urbana-Champaign, IL, USA Purpose: This study aims to characterize the histopathologic findings, clinical behavior, diagnostic findings, and treatment outcomes in canine globes affected by a syndrome associated with neovascular glaucoma of which we have proposed the name Canine Ocular Gliovascular Syndrome (COGS). Methods: The archives at COPLOW were used to identify 36 eyes with COGS. The inclusion criteria included: clusters of glial cells within the vitreous, a neovascular membrane extending from the optic nerve head or retina, and two or more histological criteria of glaucoma. Cases were randomly selected for special stain evaluations (iron, Masson trichrome, Alcian Blue, and PAS) and immunohistochemical staining for GFAP and VEGF. Clinical data, treatments, and outcomes were obtained from case records and referring veterinarian follow-up. Results: Thirty-six eyes of 35 dogs were identified with COGS. The Labrador retriever was the most predominant breed affected (20). The average age at diagnosis was 8.78 years. Average IOP at presentation was 38.7 mmHg and average time to enucleation or evisceration was 26.8 days. Most cases presented with hyphema (25) and symptoms consistent with glaucoma. The outcome in all cases was uncontrolled glaucoma and blindness in the affected eye. Bilateral COGS was confirmed histologically in one dog and suspected clinically in two additional dogs. Positive GFAP staining was found in 14 out of 16 globes. Positive VEGF staining was found in five out of six globes. Conclusions: The etiology of COGS remains unknown. It appears to have a predilection for the Labrador retriever breed. GFAP positive staining suggests astrocytic glial origin. ABSTRACT NO.: 121 Histopathology and immunohistochemistry evaluation of ocular tissues from horses (Equus caballus, Linnaeus, 1758) seropositive or seronegative for leptospirosis A. L. G. Souza,* F. Montiani-Ferreira,† B. C. Martins,* M. A. G. Silva,* R. J. S. Girio,* J. L. Laus* and L. Noronha‡ *Universidade Estadual Paulista (UNESP), Brazil, †Universidade Federal do Paraná (UFPR), Brazil, ‡Pontifı́cia Universidade Católica do Paraná (PUC), Brazil Purpose: Evaluate morphological and immune changes in ocular tissues (cornea, iris process, choroid and retina) from horses seropositive and seronegative for leptospirosis. Methods: Fifty eight globes of 29 horses from an abattoir, randomly selected, were studied using histopathology (HE) and immunohistochemistry (IHC) techniques. For all eye tissues anti-Leptospira antibodies IHC was performed. Specific antibodies were used for cornea (anti-MMP1 and anti-MMP9), ciliary process (CD74), choroid and retina (GFAP and calretinin). Serum from 29 animals as well as aqueous humor and vitreous body from 58 eyes were collected and submitted to Leptospira sp. microscopic agglutination test (MAT). Results: Fourteen animals were con- sidered positive (MAT titer higher than 80) and 15 negative). Five serovars from 26 studied from Leptospira interrogans were found: icterohaemorrhagiae (34.5%), autumalis, patoc, sentot, hebdomadis. One seronegative animal was MAT-positive for vitreous body. Analysis of HE-stained sections showed that seropositive animals for leptospirosis had corneal thickness significantly higher than the seronegative (P = 0.0347). The immunohistochemistry of corneal samples for Leptospira sp. showed higher positivity in eyes from seropositive animals but also revealed Leptospira sp. organisms even in seronegative animals (five animals). Anti-MMP9 showed significant larger area of reactivity (P = 0.0013) in sections positive for Leptospira sp. on IHC. The antibodies studied MMP1, CD74, GFAP and calretinin showed no statistically significant differences between eye samples from animals seropositive or seronegative for leptospirosis. Conclusions: Infection with Leptospira sp. may be organ-specific, since there were seronegative horses with ocular tissues positive for anti-Leptospira immunohistochemistry and a seronegative animal was MAT-positive for vitreous body. Early stages of uveitis may cause clinically imperceptible ocular signs. Leptospira sp. can cause early changes in the corneal tissues. IHC changes confirm that corneal tissues are important in the pathogenesis of the horse eye infection by Leptospira sp. and possibly equine recurrent uveitis. Supported by FAPESP – 06/54389-2. ABSTRACT NO.: 124 Diode endoscopic cyclophotocoagulation in feline glaucoma I. D. Bras and T. E. Webb Medvet Medical and Cancer Center for Pets, Worthington, OH, USA Purpose: To evaluate endoscopic diode laser system for glaucoma therapy in feline patients. Methods: Patients were selected for endoscopic cyclophotocoagulation (ECP) when intraocular pressure (IOP) was >30 mmHg, or when glaucoma was considered uncontrolled (>25 mmHg) on medical therapy. Limbal approach was used; 6/12 combined with cataract surgery, and 6/12 combined with intracapsular lens extraction. The average setting was 0.3 mW (0.25–0.325) with continuous duration; the average treated area was 230 degrees (120–320). Results: Eleven cats underwent ECP (12 eyes). 9/12 eyes had secondary glaucoma. 3/12 eyes had primary congenital glaucoma. 4/12 eyes had cataracts, 6/12 had anterior lens luxation, 1/12 had aqueous misdirection syndrome. Results: Follow-up ranged from 6 weeks to 4 years. IOP was <25 mmHg at 1h, 2h, 3h, 24h postoperatively in 83% of patients. Two weeks post-operatively, IOP was <25 mmHg in 100% of the patients. The average IOP at 6 months, 1 and 2 years was 12.38 mmHg (4–23 mmHg), 10.43 mmHg (9–19 mmHg), and 14.4 mmHg (12–18 mmHg). All patients had mild to moderate aqueous flare post-operatively, that was well controlled 4 weeks post-operatively. Complications included corneal ulcers (41.67%) and sequestrum (25%). All patients were sighted at the last examination. Glaucoma medications were eventually discontinued in 67% of patients. Conclusions: Diode ECP appears to be an effective tool in the management of glaucoma in feline patients, IOP can be controlled long term, and sight can be preserved. Incidence of ulcers/sequestra decreased when IOPs were monitored with a rebound tonometry rather than applanation tonometry. ABSTRACT NO.: 125 GNB3 is involved in cone on-bipolar cell function S. M. Petersen-Jones,* G. C. Shaw,* E. R. Ritchey,† F. Montiani-Ferreira* and A. J. Fischer† *Department of Small Animal Clinical Sciences, Michigan State University, East Lansing, MI, USA, †Department of Neuroscience, The Ohio State, Columbu, OH, USA Purpose: The RGE chick has a functional null mutation in GNB3, the gene known to encode Gb3, cone beta transducin. RGE chicks have characteristic ERG changes with elevated response thresholds and a supernormal b-wave which lacks oscillatory potentials. These abnormalities suggest that Gb3 may play a role in inner retinal function in addition to cone function. This study was undertaken to further investigate this possibility. Methods: A variety of antibodies were used in an immunohistochemical study to identify GNB3 expressing retinal neurons. The ERG of RGE chicks was dissected using the following drugs injected intravitreally: Aspartate, APB, PDA and NMDA. Dark and light adapted flash ERG and long flash ERGs were recorded pre and post injection. Results: GNB3 was found to be expressed in cones and a subset of bipolar cells in normal chicks. The Gb3 positive bipolar cells were also Islet1 positive but PKCalpha (a marker for rod bipolar cells) negative. RGE chicks lacked detectable retinal Gb3. Aspartate (blocks transmission from photoreceptor to second order neurons) eliminated the b-wave from both RGE and control chicks leaving the PIII response. In RGE chicks the PIII response had an elevated threshold but was of normal shape. APB (blocks photoreceptor to ON-bipolar cell transmission) suppressed the b-wave in control chicks. In RGE chicks it only removed an early portion of the b-wave. PDA (blocks transmission to OFF bipolar cells, horizontal cells and third order neurons) in normal chicks reduced the a-wave, enhanced the b-wave and suppressed the d-wave. In RGE chicks it removed an early negative component of the a-wave leaving an a-wave with a shallower slope and increased implicit time. It also suppressed the supernormal b-wave but enhanced the d-wave. NMDA (blocks transmission from bipolar to third order neurons) had a similar effect on the RGE chick ERG to that of PDA. Conclusions: GNB3 is expressed in cones and cone ON-bipolar cells. It appears that despite the lack of Gb3, RGE chicks still have some cone-driven responses. The function of Gb3 in cone bipolar cells is unknown but the ERG changes that result from a lack of Gb3 suggests that it may act to enhance and lengthen the duration of signal transduction within the bipolar cell. Altered cone and cone ON-bipolar cell function does not fully explain all the differences in the ERG waveform of RGE chicks. The ERG contributions from other retinal neurons are also abnormal and further studies are required to explain these changes. Supported by MSU Genetic Research Fund. ABSTRACT NO.: 126 Diagnosing Blastomycosis dermatitidis using a commercial urine antigen test Jo-Ann McKinnon, David Whitley and Dan Betts Department of Veterinary Clinical Sciences, Iowa State University, Ames, Iowa Purpose: To evaluate the clinical effectiveness of a commercial urine antigen testa as a diagnostic tool in suspected blastomycosis infection in dogs. Methods: Case information of all canine patients that were presented to the Lloyd Veterinary Medical Center College of Veterinary Medicine, Iowa State University from October 2006 to July 2009 for suspected blastomycosis were reviewed retrospectively, including the results of the urine antigen testa, cytology, urinalysis and histopathology. Results: Thirty patients were positive for blastomycosis from 113 cases. Twenty-six of those cases had a positive urine antigen testa. Four cases had false negative results; three cases were positive either by cytology or histology but had a negative urine antigen testa. One case originally reported as negative, was considered positive on a second urine anti- 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 408 a b s t r a c t s gen testa following ultrafiltration of the urine. One of the twenty-six positive cases was the only case positive on urine antigen testa and negative for all other diagnostic tests. Reviewing a subset of the tested cases, where the ocular system was the primary organ affected; twenty-seven cases were tested with the urine antigen test, six cases were positive and twenty-one were negative. Conclusions: In the population studied the sensitivity of the urine antigen testa was not the same as previously reported. There are patients for whom the test does not provide an accurate diagnosis. a Miravista Urine Antigen Test, Miravista Diagnostics, Indianapolis, IN, USA ABSTRACT NO.: 127 Lens – induced endophthalmitis. Histological findings in an animal model S. Iwabe,* F. Gil-Carrasco,† A. A. Rodriguez-Reyes† and F. Pineda‡ *Department of Pharmacobiology, Centro de Investigación y de Estudios Avanzados del IPN (Cinvestav), Mexico City, Mexico, †Glaucoma Service and Pathology Service from Asociación Para Evitar la Ceguera en México, Hospital Dr. Luis Sanchez Bulnes, IAP, Mexico City, Mexico, ‡Veterinary Ophthalmology Service, Mexico City, Mexico Purpose: To report the histological and inflammatory findings in an animal model of retinal induced damage using N-methyl-D-Aspartate (NMDA). Material and Methods: Forty rats (Long Evans) were treated with different substances. All rats received NMDA intravitreally to induce retinal damage, while 30 rats were divided in three groups that received BDNF intravitreally and/or phenytoin intraperitoneally to evaluate retinal ganglion cells (RGC) neuroprotection. Right eye had different treatments while left eye had saline solution and was used as control. Animals were euthanized after one and four weeks, eyes were enucleated and stained using Nissl staining (0.1% cresyl violet) to count RGC and HE to evaluate cells type. Results: During microscopical assessment of the 80 eyes, we unintentionally found eight eyes with acute endophthalmitis (1 week) and seven eyes with chronic granulomatous endophthalmitis (4 weeks), within these seven cases five eyes presented also some components of acute inflammation. Both conditions were observed in treated and untreated eyes. Conclusion: Accidental posterior lens capsule rupture during intravitreal injection exposes the lens proteins, probably causing the non-septic endophthalmitis. ABSTRACT NO.: 128 Ultra-sonographic aspects and biometry of striped owl’s eyes R. Squarzoni,* A. Antunes,* E. Perlmann,* L. Milanelo,† L. F. M. Barros* and P. S. M. Barros* *Veterinary Medical School, University of Sao Paulo, Brazil, †Parque Ecológico do Tiete’s Veterinary Ambulatory, Sao Paulo, Brazil Purpose: The Striped owl is found throughout most of South America, from Venezuela to Bolivia, Paraguay, Argentina, Uruguay and Brazil. The purpose of this study is to describe the normal eye anatomy of the Striped owl (Rhinoptynx clamator) and determine the normal values for axial intraocular measurements with B-mode ultrasonography. Materials and Methods: Twenty eight eyes of 14 apparently healthy young adult Striped owls from Parque Ecológico do Tiete’s Veterinary Ambulatory were examined. Ultrasonographic exams were performed with an Accutome ocular ultrasound unit, with a 12.5 to 15 MHz transducer. The owls were fixed unsedated in a lying position using common restrain techniques for raptors. The eyes were anesthesized with proximetacaine 0.5% drops (Anestalcon) and scanned in B-mode. Biometric measurements of width of the lens (WL), depth of the anterior (AC) and vitreous (VC) chambers, the length of the bulbus (LB) and the pecten oculi (LP) of both eyes were performed, in addition to routine sonographic imaging. The exams were performed by the same investigator (R.S.) to minimize measurement errors. Statistical analysis of data was performed using a software program. Results: All 27 eyes and its structures had a very similar appearance by ultrasound examination. The biometric and statistical findings were as follows: in the left eye, mean of LB was 23.11 ± 0.53 mm, mean of WL was 7.79 ± 0.27 mm, mean of AC was 4.13 ± 0.14 mm, mean of VC was 11.25 ± 0.22 mm and mean of LP was 5.59 ± 0.55 mm; in the right eye, the means were: LB = 23.77 ± 0.68 mm, WL = 8.14 ± 0.42 mm, AC = 4.43 ± 0.5 mm, VC = 11.31 ± 0.24 mm and LP = 5.56 ± 0.41 mm. One eye was excluded from biometry of anterior and vitreous chambers, and biometry of the lens because of cataracts. Correlation of measurements for LB and WL of the left and the right eyes was significantly different (P < 0.05). Conclusions: Striped owls were used on this study because according to the authors concern, there is no publication about this specie’s eyes until the moment. With a well trained staff, the restrain of the owls is safe and they don’t need to be sedated or anesthesized. The ultrasound is a non invasive exam to be performed on Striped owl’s eyes and provides very good data about morphology and biometry. Striped owl’s eyes have very similiar ultrasonographic aspects to other raptors eyes, but it is important to have its own specie biometric values. ABSTRACT NO.: 129 Oxidative stress and glutamate distribution in diabetic retinopathy in rats J. E. Madl, J. R. Gionfriddo, P. Serbedzija, A. Novakowski and D. N. Ishii Colorado State University, Ft Collins, CO, USA Purpose: Previous research has indicated that glutamine synthetase (GS) levels are decreased in diabetic retinas (Zeng, 2009). Similar decreases of GS in glaucoma may contribute to neuronal damage via altered glutamate distribution and oxidative stress (Chen, 2008; Gionfriddo, in press). Our current study tests if a similar relationship of GS depletion to altered glutamate distribution and oxidative stress occurs in diabetic retinopathy. Methods: Diabetes was induced in x rats by treatment with streptozotocin. Immunogold staining of retinas was used to evaluate amounts and distribution of GS, glutathione and glutamate. The density of staining for each antigen was measured in retinas of five rats that had been diabetic for 12 weeks and in five control rats. Results: Staining densities for GS in diabetic retinas were significantly decreased (P < 0.01) to 66.4 ± 2.7% of control retinas (n = 8). Total glutathione immunoreactivity, a marker for oxidative stress, was also significantly reduced (P < 0.01) to 64% ± 3.1 of control retinas. The losses of glutathione were especially prominent in neurons of the ganglion cell layer and inner nuclear layers rather than in the Muller cells. In initial studies, glutamate was increased in Muller cell processes but was not significantly reduced in neurons in diabetic retinas. Conclusions: GS loss from Muller cells and decreased levels of total glutathione were seen in 12 week diabetic rats. These changes appear similar to those seen in glaucoma in DBA/2J mice (Gionfriddo, in press) and may contribute to retinal damage. However, in contrast to glaucoma in dogs (Chen, 2008) and mice, a substantial loss of glutamate from neurons was not detected. Supported in part by CDC grant R49/CLR811509. ABSTRACT NO.: 130 Sildenafil reversibly suppresses rod-mediated ERG responses in dogs K. E. Pierce, J. T. Bartoe, J. G. Hauptman and S. M. Petersen-Jones Department of Small Animal Clinical Sciences, Michigan State University, East Lansing, MI, USA Purpose: To investigate if the suppressive effect of sildenafil on PDE6 activity causes permanent alterations of retinal function in carriers of the PRA causing PDE6A mutation (PDE6A+/)) in dogs. Methods: Three PDE6A+/) dogs were administered sildenafil citrate (Viagra; Pfizer, Inc., New York, NY, USA) at 14.3 mg/kg per os once daily for four months. Three wild-type dogs were administered an equivalent sildenafil dose for one week. Two PDE6A+/) dogs received placebo for four months. Ophthalmic examinations, fundus photography, electroretinography, and vision testing were performed regularly on all dogs during the pretreatment, treatment, and washout phases. Dark-adapted ERG intensity:response series and rod-mediated flicker responses were recorded followed by light adaptation and light-adapted intensity:response series and cone flicker responses. Naka-Rushton fitting was performed on b-wave dark-adapted intensity:response curves. Intensity required to elicit a dark-adapted b-wave criterion threshold of 20 lV was calculated. Vision testing was conducted at light intensities ranging from 0.002 to 20 cd/m2 using a device consisting of a junction box with four exit tunnels. The first-choice tunnel and exit time were recorded during repeated box exit attempts. Results: No ophthalmoscopic abnormalities were identified at any time point. Significantly decreased dark-adapted b-wave amplitudes (P = 0.037) and prolonged implicit times (P < 0.001) were noted during the treatment phase in all dogs administered sildenafil compared to dogs receiving placebo. The b-wave criterion threshold response was significantly elevated during the treatment phase in PDE6A+/) (P < 0.001) and wild-type (P < 0.001) dogs resulting in an ‘‘unmasking’’ of the scotopic threshold response (STR). Following washout all recorded ERG responses were comparable to pre-study values. No significant differences in initial tunnel choice or exit time were observed between groups in the vision-testing device. Conclusion: Sildenafil unmasked the STR and elevated the b-wave threshold in both PDE6A+/) and wild-type dogs during the treatment phase. Sildenafil administration at the dose and duration used in this study does not appear to cause permanent alteration of retinal function in PDE6A+/) dogs. Supported by The Midwest Eye Banks. ABSTRACT NO.: 131 Evaluation of rebound and applanation tonometry in normal and glaucomatous cats G. J. McLellan,*,† J. P. Kemmerling* and J. A. Kiland* *Department of Ophthalmology & Visual Sciences, †Department of Surgical Sciences, University of Wisconsin-Madison, WI, USA Purpose: To determine the accuracy and reproducibility of intraocular pressure (IOP) measurements obtained with the TonoVet and Tono-Pen XL tonometers in normal and glaucomatous cats. Methods: The anterior chambers of both eyes of three anesthetized adult cats (two glaucomatous, one normal) were cannulated with branched, 23-gauge needles. One branch of each needle was attached to a vertically adjustable reservoir, while the other branch allowed measurement of manometric IOP. IOP was increased by 5 mmHg increments from 5 to 70 mmHg, then decreased back to 10 mmHg, by raising and lowering the reservoir. At each increment/decrement, three TonoVet and three Tono-Pen XL readings were obtained for each eye by two independent observers. Triplicate IOP readings were also obtained with each tonometer, from both eyes of five normal and seven glaucomatous, unsedated cats, weekly for 8 weeks. Results: In both normal and glaucomatous cats, TonoVet readings were accurate, with acceptable precision (mean slope = 1.032 ± 0.036, r2 = 0.933 ± 0.045, y-intercept = 2.23 ± 2.19) and minimal inter-observer variability. Measurements obtained with the TonoPen XL were reproducible but inaccurate, as this tonometer consistently underestimated IOP at all but the lowest IOPs (mean slope = 0.622 ± 0.044, r2= 0.951 ± 0.036, y-intercept = 1.41 ± 0.65). Application of a manometrically-derived corrective equation dramatically improved the performance of the Tono-Pen XL relative to the TonoVet. Conclusions: The TonoVet tonometer allows accurate measurement of IOP in both glaucomatous and normal cats. Readings obtained with the TonoPen-XL are inaccurate but an estimate of actual IOP can be obtained in a clinical setting by use of an appropriate corrective equation. Support: NIH Grants K08 EY018609 and P30 EY0016665. ABSTRACT NO.: 132 A-luminol prevents decreases in glutamate, glutathione and glutamine synthetase in the retinas of glaucomatous DBA/2J mice K. S. Freeman,* J. R. Gionfriddo,† A. Groth,* V. L. Scofield,‡ K. Alyahya§ and J. E. Madl§ *Veterinary Medical Teaching Hospital, School of Veterinary Medicine, University of California, Davis, CA 95616, USA, †Department of Clinical Sciences, Colorado State University, 300 West Drake, Fort Collins, CO 80523, USA, ‡MD Anderson Cancer Research Center, Smithville, TX 78957, USA, §Biomedical Sciences, Colorado State University, 300 West Drake, Fort Collins, CO 80523, USA Purpose: To test the hypotheses that in DBA/2J mice, oxidative stress decreases glutamine synthetase (GS) levels resulting in a loss of neuronal glutamate and that the antioxidant A-luminol (GVT) decreases both oxidative stress and glutamate loss in some types of glaucoma. Methods: DBA/2J mice were separated into two groups: one not treated, and the other treated with 1% w/v GVT in the drinking water. At seven months of age, retinas were examined from five untreated DBA/2J mice, seven GVT-treated mice, and five C57BL/6 mice (negative controls). Serial 0.5 lm plastic sections were immunogold stained for glutamate, GS, and total glutathione, followed by image analysis for staining patterns and density. Results: Focal decreases in glutamate immunostaining were common in the inner nuclear layer (INL) of DBA/2J retinas, but not in C57BL/6, nor GVT-treated DBA/2J retinas. Decreases in glutathione and GS immunostaining were found in DBA/2J retinal regions where neuronal glutamate immunostaining 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409 abstracts 409 was reduced. Retinas from GVT-treated DBA/2J had no significant decreases in INL levels of glutamate, glutathione, or GS. Conclusions: Retinas of dogs with primary glaucoma are reported to have focal depletion of neuronal glutamate. In DBA/2J mice, similar changes occur prior to development of clinical disease. In these focal glutamate-depleted regions, levels of glutathione and GS are also reduced, consistent with the hypothesis that oxidative stress contributes to retinal changes in glaucoma. The ability of GVT, an antioxidant, to inhibit retinal abnormalities in DBA/2J mice provides further support for this hypothesis. Supported by Colorado State University College Research Council Grant R49/CLR 811509. GVT supplied by Bach Pharma, North Andover, MA. ABSTRACT NO.: 133 Gene therapy in the second eye of RPE65 deficient dogs also improves retinal function M. J. Annear,* J. T. Bartoe,* S. E. Barker,‡ A. J. Smith,‡ P. G. Curran,† J. W. Bainbridge,‡ R. R. Ali‡ and S. M. Petersen-Jones* *Department of Small Animal Clinical Sciences, †Center for Statistical Consulting, Michigan State University, East Lansing, MI, USA, ‡Division of Molecular Therapy, UCL Institute of Ophthalmology, London Purpose: To evaluate whether prior subretinal gene therapy in the RPE65-/- dog using an AAV2/2 construct containing the human RPE65 gene results in immune responses that interfere with rescue when the contralateral eye is similarly treated. Methods: Nine RPE65-/- dogs underwent subretinal injection of an AAV2/2 vector containing the human RPE65 gene coding region driven by the human RPE65 promoter. Subretinal injection of the same vector construct was performed in the contralateral eye 90–180 days after the first injection. Rescue of retinal function was assessed by electroretinography and vision testing. A dark-adapted intensity:response series and rod flicker responses were recorded. Following light-adaptation single flash and flicker responses were recorded. To assess rod and cone rescue ERG responses at 1 cdS/m2 (below threshold for an untreated RPE65-/- dog) and 33 Hz flicker responses respectively were measured. A vision testing apparatus that assessed a subject’s ability to see an open exit tunnel was used to quantitatively measure visual function under differing light levels. Serum was collected to assess immune response to AAV2 (serum neutralizing antibodies) and RPE65 protein (IgM and IgG levels by ELISA). Statistical analysis was performed with significance set at P £ 0.05. Results: Rod and cone rescue was present in 16 of 18 injected eyes, and remained static up to 2 years post treatment. There was no significant difference in rod or cone response between the first or second injected eyes at any time point. Vision testing found significant improvement in both ‘time to exit’ and ‘first correct choice of exit’ relative to pre-treatment values. There was no significant difference between first or second treated eyes. A circulating immune response to the vector and RPE65 protein was detected but the level of response did not correlate with degree of rescue in the second eye. Conclusion: Successful rescue using AAV2 gene therapy in the second eye is possible in the RPE65-/- dog. This has important implications for treatment of human Leber congenital amaurosis type II patients. Supported by the British Retinitis Pigmentosa Society. ABSTRACT NO.: 134 Prevalence of Leptospira infections and positive PCR tests in horses with chronic or recurrent uveitis in Louisiana M. A. Kubai, R. T. Carter and E. S. Storey School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA, USA Purpose: To investigate the prevalence of Leptospira infections within the eyes of horses presented to Louisiana State University for chronic or severe uveitis. Methods: Horses were sedated to allow the aqueous humor to be sampled aseptically for diagnostic purposes at presentation, or vitreous was harvested for Leptospira PCR during a transcleral vitrectomy for the treatment of unresponsive equine uveitis. In addition to Leptospira PCR, samples of the same intraocular fluids were submitted to a diagnostic laboratory for Leptospira cultures, and titers as sample volume permitted. Leptospira culture and transport media were obtained in advance from the laboratories. Results: Six out of eleven (55%) horses diagnosed with uveitis had positive PCR results for Leptospira. Two of the horses with positive Leptospira PCR tests (18%) also had positive cultures for Leptospira. Serology was submitted for 10 of the 11 horses and all horses tested had positive serum, vitreous and/or aqueous titers for Leptospira. Conclusions: The population of horses that presented to LSU with uveitis had a high prevalence of Leptospira based on PCR results obtained from intraocular fluids (aqueous and vitreous). Infectious causes of equine uveitis including Leptospira should be considered in the etiologic differentials for equine uveitis, but the diagnosis of these intraocular infections was not aided by serology and required specific, invasive sampling of the ocular fluids. ABSTRACT NO.: 135 Keratomycosis caused by Curvularia sp. in an Asian elephant (Elephas maximus): diagnosis and treatment A. B. Marlar, S. T. Ferrell and N. P. Lung Fort Worth Zoological Association, Forth Worth, TX, USA A 35-year old, female Asian elephant (Elephas maximus) presented acutely with ocular pain and a focal, perilimbal corneal opacity. Initial examination findings included epiphora, marked conjunctival hyperemia, inferior corneal edema and limbal vascularisation with pigmentation. Inflammatory infiltrates were present in the central portion of the lesion. At presentation, the corneal lesion stained negative with fluorescein dye. Initial corneal bacterial culture yielded S. hyicus and C. striatum. Corneal scrapings were achieved with the animal in lateral recumbency. Corneal cytology was suggestive of keratomycosis. Fungal culture from the lesion was positive for Curvularia sp. Medical management included: topical EDTA plasma, antibiotics, antifungal agents, intermittent debridement and oral anti-inflammatory therapy. Treatment continued for eight weeks at declining frequencies (from q 2 h to q 12 h) until the lesion appeared completely resolved. This case illustrates that while there are challenges associated with performing ophthalmic exams and diagnostics in megavertebrate species, efforts to do so are important to help obtain accurate diagnosis and aid in directing therapy. The challenges and techniques facilitating successful medical management in this case will be discussed. The authors would like to acknowledge the generous support of Alcon Laboratories in assisting with the management of this case and the help and dedication of the elephant staff at the Fort Worth Zoo. ABSTRACT NO.: 136 The dark adaptation curve of the horse G. Ben-Shlomo, C. Plummer, K. Barrie and D. Brooks College of Veterinary Medicine, Department of Small and Large Animal Clinical Sciences, University of Florida, FL, USA Purpose: There is a lack of information in the literature regarding the electroretinogram (ERG) of the horse. Despite a clinical need, the dark adaptation curve of the horse has not been investigated to date. The goal of this work is to study the dark adaptation curve of the normal horse. Methods: The ERG responses were recorded from five healthy female ponies using a contact lens electrode and the mini Ganzfeld ElectroRetinoGraph. The horses were sedated intravenously with detomidine (0.015 mg/kg), an auriculopalpebral nerve block was then performed, and the pupil was fully dilated. The ERG was recorded in response to a low intensity stimulus (10 mcd.s/m2) that was given at times (T) T = 5, 10, 15, 20, 25, 30, 40, 50 and 60 minutes of dark adaptation, and off-line analysis was then performed. Results: Mean b-wave amplitude of the full-field ERG did not changed significantly between T = 5 and T = 15 (increase of 8.7%, P = 0.63). A significant increase (26.3%) of b-wave amplitude was noted between T = 15 to T = 20 (P < 0.05). The b-wave amplitude was peaked at T=25, however there was no statistical significance between T = 20 and T = 25. The b-wave amplitude remained with no significant changes until the end of the study at T = 60 (P > 0.49). Conclusions: Evaluation of horse rod function or combined rod/cone function by means of full field ERG should be performed after a minimum 20 minutes of dark adaptation. ABSTRACT NO.: 137 Topical Kinostat ameliorates the development and progression of cataracts in dogs with diabetes mellitus I. D. Bras,* T. R Webb,* K. Ketring,† M. Wyman*,‡,§ and P. F. Kador‡,– *MedVet Medical Center for Pets, Worthington, OH, USA, †All Animal Eye Clinic, Cincinnati, OH, USA, ‡Therapeutic Vision Inc., Omaha, NE, USA, §College of Veterinary Medicine The Ohio State University, Columbus, OH, USA, –College of Pharmacy University of Nebraska Medical Center, Omaha, NE, USA Purpose: Cataract formation is a hallmark of diabetes mellitus (DM) in dogs. Approximately 75% of diabetic dogs develop cataracts within 1 year of the time of diagnosis. Since the aldose reductase catalyzed formation of sorbitol in the lens has been experimentally demonstrated to initiate cataracts in diabetic animals, the purpose of this study was to investigate whether the topical administration of the aldose reductase inhibitor Kinostat can ameliorate the onset or progression of cataracts in naturally occurring diabetic dogs. Methods: Forty dogs, newly diagnosed with DM and with minimal lens changes were enrolled in a prospective, masked pilot study. Dogs were randomly assigned a coded vial containing either Kinostat or vehicle (placebo), with the contents of the vial (drug or placebo) masked from the examiner. Twenty-eight dogs received Kinostat, and 12 dogs received placebo. Owners were instructed to administer the agent OU TID for 1 year, and were instructed to record each time of administration in order to ensure compliance. Complete ophthalmic examinations were performed prior to enrolling in the study, and then at 1, 2, 3, 6, 9, and 12 months after. Digital images were taken after dilation at each time interval. Complete blood chemistries, HbA1C, and Kinostat blood levels were obtained at 1 year. Results: Cataract formation and progression were observed in 10/12 (83%) of the dogs receiving placebo with seven dogs developing mature cataracts, two developing cortical opacities and one developing equatorial vacuoles. Two dogs (16%) in the placebo group did not show evidence of cataract formation at the last follow up. In contrast, cataract formation in the Kinostat group was significantly (P = 0.0016) inhibited with 15/28 (53.6%) of dogs receiving Kinostat not showing evidence of cataract development at the last follow up. Of the Kinostat treated dogs, eight developed anterior equatorial vacuoles, two developed incipient anterior cortical cataracts, and four developed mature cataracts by 12 months. Of the 20 KinostatTM treated dogs remaining on study at time periods ranging from 14 to 22 months, 13 have no lens changes, six have cortical vacuoles and one has a cortical opacity. This is significantly different (P = 0.0001) from the seven remaining placebo dogs where one has no lens changes, one has cortical vacuoles, two have cortical opacities and three have mature cataracts. HbA1C values were not significantly different (P = 0.369) between the Placebo (6.7 ± 0.95) and KinostatTM (7.7 ± 1.8) groups. Conclusion: Topical Kinostat is beneficial in arresting the onset and/or progression of cataracts in dogs with diabetes mellitus. Drs. Wyman and Kador have financial interest in this study and are employees of Therapeutic Vision. Supported by NIH SBIR 1R43EY018013-01A1. 2009 American College of Veterinary Ophthalmologists, Veterinary Ophthalmology, 12, 390–409