Corneal transplantation for inflammatory keratopathies in the horse: visual outcome in 206 cases (1993-2007)

Objective To evaluate the visual outcome of three techniques of corneal transplantation surgery in treating severe inflammatory keratopathies in the horse. Design Retrospective medical records study. Animals studied Medical records of 206 horses that received corneal transplantation surgery at the University of Florida Veterinary Medical Center from 1993 to 2007 were reviewed. Procedure Data collected from the medical records included signalment, types of ocular lesions, type of transplant surgery performed, length of follow‐up, complications, and visual outcomes. Results Full thickness penetrating keratoplasty (PK) was performed in 86 horses for melting ulcers, iris prolapse/descemetoceles, and medically nonresponsive full thickness stromal abscesses (SA). Posterior lamellar keratoplasty (PLK) and deep lamellar endothelial keratoplasty (DLEK) are split thickness penetrating keratoplasties that were utilized for medically nonresponsive deep stromal abscesses (DSA) in 54 and 66 eyes, respectively. The most common postoperative surgical complication was graft rejection and varying degrees of graft opacification. Wound dehiscence and aqueous humor leakage was also a common postoperative problem. A positive visual outcome was achieved for PK, PLK, and DLEK in 77.9%, 98.1%, and 89.4%, respectively. Conclusions Corneal transplantation is a tectonically viable surgery in the horse with an overall success rate of 88.5% in maintaining vision when treating vascularized and infected corneal disease in the horse.     

Deep lamellar endothelial keratoplasty in 10 horses

Objective  To describe and evaluate a surgical technique utilized for the therapy of deep corneal stromal abscesses (DSA) in horses. The DSA is excised and replaced with a partial thickness corneal lamellar allograft.
Methods  A retrospective clinical study describing the indications for the surgical technique utilized and the outcomes of this procedure in 10 eyes of 10 horses.
Results  Each affected eye had a discrete DSA within the posterior stroma. An initial partial thickness semicircular corneal incision was made at the limbus, followed by anterior stromal lamellar dissection over the lesion. After excision of the DSA and replacement with a larger diameter split-thickness donor button, the anterior stroma was replaced into its original position and the initial corneal incision was repaired. All of the animals that underwent deep lamellar endothelial keratoplasty (DLEK) procedure healed appropriately and with subjectively less postoperative scarring and complications than previously described surgical approaches to DSA.
Conclusions  This procedure is an effective technique for surgical removal of DSA in horses and, in most cases, results in a visual and cosmetically acceptable globe. The advantages of this technique compared to other surgical approaches to DSA are the peripheral location of the incision, shortened anesthesia times, the resultant minimal scarring and shorter healing times associated with DLEK.     

Carbon Dioxide Laser Keratectomy as a Treatment Option for Equine Corneal Stromal Abscessation: A Comparison to Lamellar Keratectomy and Penetrating Keratoplasty

Carbon dioxide (CO₂) laser keratectomy was compared to lamellar keratectomy (LK) and penetrating keratoplasty (PK) as a potential treatment option for equine corneal stromal abscessation. The medical records of 35 client owned horses undergoing surgical management of stromal abscessation were reviewed. Among those 17 underwent LK, 3 PK and 15 CO₂ laser keratectomy. Healing of the stromal abscess was achieved in all treated eyes. Those horses undergoing CO₂ laser keratectomy demonstrated few complications, and the time to healing and cosmetic outcome was comparable to those horses undergoing LK and PK.     

Ulcerative keratitis caused by beta-hemolytic Streptococcus equi in 11 horses

Purpose To describe 11 clinical cases of ulcerative keratitis in horses associated with beta-hemolytic Streptococcus equi in Florida, USA. METHODS: Retrospective clinical study (1996-99). RESULTS: Beta-hemolytic Streptococcus equi was cultured from 11 horses with deep ulcers, descemetoceles or iris prolapse (n = 8), a suture abscess found with a penetrating keratoplasty for a stromal abscess (n = 1), and ulceration that developed following keratectomy/irradiation for corneal squamous cell carcinoma (n = 2). Beta-hemolytic Streptococcus equi subspecies zooepidemicus was found in 10 eyes and subspecies equi in one. Marked signs of uveitis including miosis and hypopyon were present in 8/11 (72.7%) eyes. Keratomalacia was severe in all eyes. The mean diameter of the ulcers associated with beta-hemolytic Streptococcus was 10.2 +/- 6.1 mm. Eight of the eyes required conjunctival flap surgery (four grafts dehisced) and one eye corneal transplantation. Two eyes were treated with medication only. Isolate sensitivity to antibiotics included ampicillin (6/11), bacitracin (11/11), cephalothin (11/11), chloramphenicol (11/11), gentamicin (5/11), polymyxin B (2/11), and tobramycin (1/11). All isolates were resistant to neomycin. The average healing time was 44.7 +/- 26.7 days. The visual outcome was positive in 8/11 eyes, and the globe retained in 9/11 eyes. CONCLUSIONS: Although Gram-positive bacteria predominate in the normal conjunctival microflora of horses throughout the world, Gram-negative bacteria and fungi are more often isolated from equine ulcers. Beta-hemolytic Streptococcus spp. are associated with a very aggressive ulcerative keratitis with the capability to digest conjunctival graft tissue. Clinical signs are pronounced. Aggressive surgical and intensive medical therapy with topical antibiotics and protease inhibitors is indicated.     

Equine corneal stromal abscesses: An evolution in the understanding of pathogenesis and treatment during the past 30 years

The last 30 years have seen many changes in the understanding of the pathogenesis and treatment of equine corneal stromal abscesses (SAs). Stromal abscesses were previously considered an eye problem related to corneal bacterial infection, equine recurrent uveitis, corneal microtrauma and corneal foreign bodies in horses. They were more commonly diagnosed in horses living in subtropical climatic areas of the world. Therapeutic recommendations to treat equine SAs were historically nearly always a medical approach directed at bacteria and the often associated severe iridocyclitis. Today the pathogenesis of most equine SAs appears to be more often related to fungal inoculation of the anterior corneal stroma followed by posterior migration of the fungi deeper into the corneal stroma. There is also now an increased incidence of diagnosis of corneal SAs in horses living in more temperate climates. Medical and surgical treatments are now directed towards elimination of fungal and bacterial infections, reduction and replacement of diseased corneal stroma, and suppression of iridocyclitis. If the abscess and anterior uveitis do not respond satisfactorily to medical therapy, full thickness or split thickness lamellar keratectomy to remove the fungal hyphae and diseased stroma, followed by transplantation of healthy corneal allografts has a high rate of success in speeding healing and preserving sight. This paradigm shift in the ability to diagnose and institute therapy for corneal SAs in horses has evolved over the last 30 years, and is the focus of this paper.     

Corneal stromal abscess in a horse

A thoroughbred yearling presented with a focal, yellow, midstromal corneal opacity with concurrent iridocyclitis which was consistent with a corneal stromal abscess. When continued, appropriate, medical therapy failed to improve the patient's condition, penetrating keratoplasty was performed for diagnosis and therapy. Histopathology showed that the deep corneal stroma and Descemet's membrane were severely infiltrated with necrotic neutrophils and numerous, intralesional fungal hyphae. Culture was negative. Bacteria were not isolated, consistent with a fungal corneal stromal abscess. The resultant corneal scar did not interfere with the horse's racing career.     

Profiles of matrix metalloproteinase activity in equine tear fluid during corneal healing in 10 horses with ulcerative keratitis

OBJECTIVE: Levels of tear film matrix metalloproteinases (MMPs) activity are significantly elevated in horses with ulcerative keratitis and contribute to the excessive breakdown of stromal collagen. Changes in the amount of proteolytic activity in horse tear film during corneal healing and stromal remodeling have not yet been reported, but we hypothesize they should decrease. In the present study we analyzed serial tear fluid from horses with ulcerative keratitis to identify any changes in MMP activity during corneal healing and stromal remodeling. PROCEDURES: Samples of tear fluid were obtained from both eyes of 10 horses with ulcerative keratitis on the day of admission (day 1) at the hospital and then at various time points until complete healing of the cornea. Tear film MMP2 and MMP9 activity was determined by quantitative gelatin zymography. In all cases medical treatment included topical applications of equine serum, antibiotics, atropine and systemic administration of anti-inflammatory drugs. Surgical procedures were performed in several cases on day 2 in addition to the medical treatment. RESULTS: The mean total MMP activity (+/- SD) measured in relative standard units (RSU) in the tear fluid of the ulcerated eye (2.44 +/- 1.44) of the 10 horses was significantly higher than the mean in the contralateral eye (0.81 +/- 0.68) (P = 0.006), on the day of admission at the VMTH. The mean MMP activity in these ulcerated eyes significantly decreased (-82.4%) between the first day of admission and the day when the ulcer had completely healed (P = 0.0002). The activity level in the healed eye (0.43 +/- 0.17) was not significantly different to the one in the contralateral eye (0.36 +/- 0.18) on the day of complete corneal healing (P = 0.374). The level of MMP activity in the contralateral eye also decreased from 0.81 +/- 0.68-0.36 +/- 0.18 but this decrease (56%) was not significant (P = 0.069). CONCLUSIONS: Ulcerative keratitis in horses is associated with initially high levels of tear film proteolytic activity that decrease as the ulcers heal. The success of medical and surgical treatment of the corneal ulcers is reflected by the enzyme activity in tears. In horses successful treatment does lead to a rapid reduction in tear film proteolytic activity that corresponded with the improvement in the clinical signs of corneal ulceration. Measurement of MMP activity in the tear film might represent a way to monitor the progression of corneal healing in horses with ulcerative keratitis.     

The use of porcine small intestinal submucosa for the repair of full-thickness corneal defects in dogs, cats and horses

OBJECTIVE: To evaluate the efficacy of using a porcine small intestinal submucosa (SIS) graft covered by a conjunctival flap for the surgical repair of full-thickness corneal wounds in dogs, cats and horses. PROCEDURE: All records dating from August 1999 to February 2003 from Purdue University Veterinary Teaching Hospital of patients that had undergone ophthalmic surgical procedures and received a SIS corneal graft for a full-thickness lesion were reviewed. Fifteen cases were identified including six dogs, two cats and seven horses. Requirements for inclusion in this study were that SIS was used as a corneal graft in a full-thickness corneal defect and that the graft was completely covered with a conjunctival flap. RESULTS: Of the 15 cases, one canine patient had received SIS following removal of an epibulbar melanocytoma. The remaining five canine patients had undergone this surgical procedure for the repair of corneal perforation. The two feline patients had been presented for corneal perforation following chronic ulceration. One equine patient had been presented for a deep melting ulcer, three for stromal corneal abscesses, and three for corneal perforations. Complications encountered postoperatively included aqueous leakage, conjunctival flap dehiscence, synechia, cataract and fibrin in the anterior chamber. Fourteen out of 15 patients were visual at the final re-evaluation. CONCLUSION: SIS is an inexpensive, easy-to-handle biomaterial that appears to be suitable for the repair of full-thickness corneal wounds in dogs, cats and horses. Results of our study support the conclusion that this relatively new product is an effective alternative to traditional implantation materials utilized in veterinary ophthalmology.     

Clinical findings and outcomes of ulcerative keratomycosis in 30 horses in the mid‐Atlantic United States (2006–2007)

The purpose of this study was to determine the clinical course and outcome associated with keratomycosis in horses in the mid‐Atlantic USA. Records of horses diagnosed with keratomycosis at New Bolton Center from November 2006 to November 2007 with positive fungal culture were retrospectively studied. Neither horses with ulcerative keratitis and a negative fungal culture nor those with stromal abscesses were included. Subject details, history, clinical findings, therapy and outcome were recorded. Thirty horses fitted both inclusion criteria (diagnosis of keratomycosis and positive corneal fungal culture). Fourteen of 30 cases occurred during summer. Aspergillus was the most commonly cultured fungal genus (17/30, or 57%) followed by Alternaria (4/30). Seventeen horses had positive bacterial and fungal cultures. Fifteen of 30 horses were treated surgically by a keratectomy and amnion (8) or conjunctival (7) graft. Itraconazole was the most common topical anti‐fungal therapy and was utilised in 25/30 horses. Globe survival was 97% (29/30). All surviving globes had a positive menace response and were visual at the last examination. It was concluded that in the mid‐Atlantic USA, fungal keratitis is common, has the highest incidence in summer, and is usually associated with a positive outcome. Aspergillus may be a relatively more common corneal pathogen in this region than elsewhere in the USA. Surgical cases were more likely to have fungal hyphae identified on cytology and tended to be hospitalised longer than medical cases. There was no apparent association between surgical disease and all other patient, organism and treatment variables.     

Immune-mediated keratitis in horses: 19 cases (1998-2004)

OBJECTIVE: The purpose of this study is to describe clinical and histologic findings, treatment, and outcome of horses with suspected immune-mediated keratitis (IMMK). DESIGN: Retrospective study. ANIMALS: Nineteen horses that presented to NCSU-VTH from 1998 to 2004 with IMMK. Procedures Information retrieved from the medical records included signalment, duration of clinical signs, therapy prior to initial examination, ophthalmic abnormalities, diagnostics performed, therapy instituted, and long-term vision. RESULTS: Nineteen horses (22 eyes) were diagnosed with IMMK. Three distinct clinical groups were identified based on the depth of the lesion in the cornea: superficial stromal (n = 11 eyes), midstromal (n = 6 eyes), or endothelial (n = 5 eyes). Horses ranged from 5 to 19 years of age, with a mean age +/- SD of 11.9 +/- 3.6 years. Eleven horses had 12 months or greater duration of clinical signs of corneal disease prior to referral. Overall there was a mean duration of 11.8 +/- SD 8.3 months. Superficial stromal keratitis appeared as a superficial stromal cellular infiltrate with diffuse vascularization. Midstromal keratitis appeared as midstromal cellular infiltrate with mild, surrounding corneal edema and vascularization. Endothelial disease appeared as endothelial cellular infiltrate with diffuse corneal edema. In all types of IMMK, signs of uveitis or severe discomfort were not observed. CONCLUSIONS AND CLINICAL RELEVANCE: Horses with superficial IMMK responded to topical medical therapy, but responded best to surgical removal of the lesion. Horses with midstromal keratitis responded to topical cyclosporine therapy. Endothelial disease was the least amenable to therapy.     

Rose bengal positive epithelial microerosions as a manifestation of equine keratomycosis

Purpose To describe the clinical appearance of corneal epithelial cell microerosions associated with keratomycosis in the horse. METHODS: Retrospective clinical study. RESULTS: Multifocal, punctate, superficial corneal opacities with positive rose bengal retention were noted in six horses with presumed 'viral keratitis'. Faint fluorescein staining was also present in three cases. Equine herpesvirus tissue culture inoculation was negative for a cytopathic effect in three cases. Aspergillus (n = 3), Curvularia (n = 1), and an unidentified fungus (n = 1) were cultured in five horses, and hyphae found on corneal cytology from the sixth. Mixed bacterial infections were present in three eyes. The eyes of two horses with Aspergillus progressed to deep melting corneal ulcers that required surgical therapy. The microerosions remained superficial, but persistent in the other four eyes. Natamycin was utilized topically in all six horses. Transmission electron microscopy from case 6 revealed mucin layer disruption, an intact corneal epithelial cell layer, and fungal attachment to degenerating epithelial cells. The visual outcome was positive in all six horses, although healing was prolonged (48.5 +/- 14.5 days on average in the horses with no surgery; 62 days on average in the two horses that required surgery). CONCLUSIONS: Complete removal or full-thickness penetration of the corneal epithelial cell barrier may not be necessary to allow fungal adherence and initiation of keratomycosis in the horse. Prior to colonization and invasion of the horse cornea, fungi may induce changes in the mucin layer of the tear film that result in or are associated with rose bengal positive microerosions of the superficial corneal epithelium. Horses with painful eyes, and eyes with superficial, multifocal corneal opacities should have their corneas stained with both fluorescein and rose bengal as fungal microerosions may stain weakly, or not at all, with fluorescein, and may thus be mistaken for presumed 'viral keratitis' of the horse.     

A keratoprosthesis prototype for the dog

Objective  To describe the technique for implantation of a novel keratoprosthesis (KP) prototype and evaluate its application for the treatment of corneal blindness in dogs.
Animals studied  Seven dogs, all of them being clinically blind before surgery as a result of severe corneal endothelial disease (5/7) or chronic superficial keratitis (2/7) that were unresponsive to prior therapy.
Procedures  A silicone KP was implanted unilaterally, just anterior to Descemet's membrane, after creating a stromal pocket by deep stromal lamellar dissection.
Results  Implantation of the KP was accomplished without complication in six of seven operated dogs. In the remaining case, an intra-operative complication (perforation of Descemet's membrane) was associated with extrusion of the KP 8 weeks postoperatively. All operated eyes regained limited vision after surgery. Three to six months after implantation purulent keratitis occurred in all five eyes with endothelial disease, necessitating surgical removal of the KP 6 months postoperatively in 5/7 eyes.
Conclusions  This KP prototype shows promise as a treatment for certain blinding corneal diseases. However, changes in the design of this KP, allowing improved stromal integration, will be necessary before its clinical application can be approved.     

Effect of topical administration of epidermal growth factor on healing of corneal epithelial defects in horses

OBJECTIVE-To characterize healing of corneal epithelial defects in horses and to evaluate the ability of epidermal growth factor (EGF) to modulate rate of corneal epithelial healing in horses. SAMPLE POPULATION: 20 eyes in 12 adult horses. PROCEDURE: Corneal epithelial wounds were created by mechanically debriding the limbus. Corneal healing was recorded for 3 treatment groups: 50 microg of EGF/ml (n = 5 eyes), 5 microg of EGF/ml (7), and PBS solution (8). Corneal healing was recorded once daily after instillation of fluorescein stain by use of photography and calculating the area of the wound, using imaging software. RESULTS: After corneal debridement, re-epithelialization was rapid and progressed in a linear fashion for the first 5 to 7 days after surgery in all groups. After that period, rates of healing decreased. A profound increase in the degree of inflammation, neovascularization, melanosis, and scarring was observed in eyes treated with the high dose of EGF (50 microg/ml), but there was not a statistical difference in mean healing time or in mean decrease in radius during the linear phase between the control and either EGF treatment groups. However, for all 8 horses in which both eyes were debrided, the first eye healed significantly faster than the second eye, regardless of treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Beneficial effects of topical administration of a high dose of EGF for acceleration of healing of corneal defects in eyes of horses are outweighed by the intensity of the associated inflammatory response.     

Grid keratotomy as a treatment for superficial nonhealing corneal ulcers in 10 horses

OBJECTIVE: To describe the clinical symptoms of 10 cases of superficial nonhealing corneal ulcers in horses and to evaluate the results of grid keratotomy in these patients. DESIGN: Retrospective study. ANIMALS: Ten horses with superficial nonhealing corneal ulceration in one eye. PROCEDURE: The signalment, history and clinical symptoms are reported of 10 patients with superficial nonhealing corneal ulcers during the period from August 2003 to February 2005. Grid keratotomy was performed in all cases. In addition, the surgical procedure of grid keratotomy and response to therapy are described. RESULTS: Horses generally responded well to grid keratotomy (eight cases healed after one grid keratotomy and one horse healed after a second grid keratotomy; one case was not available for follow-up). Only 2/10 had discomfort after treatment and only 2/9 had some degree of scarring after treatment. The healing time, which was known in seven cases, averaged 8.4 days (+/- SD 4). CONCLUSIONS AND CLINICAL RELEVANCE: Grid keratotomy is an appropriate option for treatment of superficial nonhealing corneal ulcers in horses. The procedure is simple to perform. It generally induces a rapid and uncomplicated healing of the cornea. Only in a limited number of cases does a small amount of scarring occur.     

Amniotic membrane transplantation for corneal surface reconstruction after excision of corneolimbal squamous cell carcinomas in nine horses

OBJECTIVE: The purpose of this study was to evaluate the usefulness and effectiveness of permanent amniotic membrane transplantation as an adjunctive treatment to superficial keratectomy alone or combined with strontium-90 irradiation for treatment of equine corneolimbal squamous cell carcinoma (SCC) to decrease corneal scarring and recurrence rate. STUDY: The retrospective case study included 11 horses (n = 12 eyes) diagnosed and treated for ocular SCC that involved the limbus and cornea. Nine of those horses (n = 9 eyes) were treated between 2002 and 2006, with superficial lamellar keratectomy alone or combined with strontium-90 irradiation and followed by placement of a permanent amniotic membrane graft in the surgical defect. The level of scarring (i.e. the clarity of the cornea) resulting with the use of amniotic membrane was subjectively compared to cases where a permanent bulbar conjunctival graft was performed following keratectomy combined with strontium-90 irradiation or cryotherapy (n = 3 eyes). Recurrence was defined as the postoperative and postirradiation regrowth of SCC in the same site and globe. RESULTS: The nine horses that received an amniotic membrane graft after keratectomy alone or combined with irradiation showed a minimal level of scarring in a cornea that regained a greater transparency in comparison to the horses that were treated with a bulbar conjunctival graft. All of the horses that received an amniotic membrane graft had 226 +/- 218 days of follow-up without tumor recurrence (mean +/- SD), ranging from 21 days to 778 days. CONCLUSIONS: The combination of superficial keratectomy alone or associated with beta-irradiation and permanent amniotic membrane transplantation is an effective treatment of corneal or corneolimbal SCC in horses. The placement of an amniotic membrane material represents an alternative surgical procedure to bulbar conjunctival grafts, especially if there is a lack of bulbar conjunctiva tissue available after tumor resection or if a particularly large corneal resection is necessary. The amniotic membrane is incorporated into the corneal defect and seems to create noticeably much less scarring than a corneal defect covered by bulbar conjunctiva.     

Efficacy of a long-acting formulation of ceftiofur crystalline-free acid for the treatment of naturally occurring infectious bovine keratoconjunctivitis

OBJECTIVE: To evaluate the efficacy of ceftiofur crystalline-free acid (CCFA) administered into the posterior aspect of an ear for treatment of corneal ulceration associated with naturally occurring infectious bovine keratoconjunctivitis (IBK). ANIMALS: 78 beef calves located at Sierra Foothills Field Station (SFS) and 52 calves located at a commercial dairy (CD). All calves were from 3 to 9 months old. PROCEDURE: At each site, calves were randomly allocated to 1 of 2 treatment groups by use of a block design determined by corneal ulcer size. A single dose of CCFA (6.6 mg of ceftiofur equivalents/kg, s.c.) was administered into the posterior aspect of a pinna. A second group of calves received a single dose of vehicle (0.03 mL/kg, s.c.; controls). Corneal ulcers were photographed, and clinical signs were assessed in calves every 3 to 4 days for 21 days. RESULTS: A positive treatment effect was detected at SFS. Results at the CD were inconclusive because ulcer healing occurred rapidly in control and CCFA-treated calves. At SFS, treatment with CCFA resulted in shorter mean healing times, smaller corneal ulcer surface area measurements, amelioration of ocular discharge and photophobia, and a 50% increase in the percentage of calves healed by day 14. After adjustment for initial corneal ulcer size, treatment with CCFA resulted in a 4-fold increase in the odds of corneal ulcer healing by day 14, compared with controls. CONCLUSIONS AND CLINICAL RELEVANCE: A single dose of CCFA administered into the posterior aspect of a pinna had a positive treatment effect against naturally occurring IBK in calves with corneal ulcerations.     

Isolation and cultivation of equine corneal keratocytes, fibroblasts and myofibroblasts

Objective  To establish an in vitro model for the investigation of equine corneal wound healing. To accomplish this goal, a protocol to isolate and culture equine corneal keratocytes, fibroblasts and myofibroblasts was developed.
Animal material  Equine corneal buttons were aseptically harvested from healthy research horses undergoing humane euthanasia for reasons unrelated to this study. Slit-lamp biomicroscopy was performed prior to euthanasia by a board-certified veterinary ophthalmologist to ensure that all samples were harvested from horses free of anterior segment disease.
Procedure  Equine corneal stroma was isolated using mechanical techniques and stromal sub-sections were then cultured. Customized media at different culture conditions was used to promote growth and differentiation of corneal stromal cells into keratocytes, fibroblasts and myofibroblasts.
Results  Cell culture techniques were successfully used to establish a method for the isolation and culture of equine corneal keratocytes, fibroblasts and myofibroblasts. Immunohistochemical staining for alpha-smooth muscle and F-actin was used to definitively differentiate the three cell types.
Conclusion  Equine corneal stromal keratocytes, fibroblasts and myofibroblasts can be predictably isolated and cultured in vitro using this protocol.