Evaluation of matrix metalloproteinase concentrations in precorneal tear film from dogs with Pseudomonas aeruginosa-associated keratitis

OBJECTIVE: To evaluate the changes in concentrations of matrix metalloproteinase (MMP)-2 and MMP-9 in the precorneal tear film of dogs with Pseudomonas aeruginosa-associated keratitis during corneal healing and stromal remodeling. ANIMALS: 10 dogs with unilateral P aeruginosa-associated keratitis and 10 clinically normal dogs. PROCEDURES: Precorneal tear film samples were collected from both eyes of 10 dogs with unilateral P aeruginosa-associated keratitis on the day of admission to the hospital and then at various time points until complete healing of the cornea was achieved. Precorneal tear film samples were also collected from both eyes of 10 clinically normal adult dogs (control group). Concentrations of MMP-2 and MMP-9 in precorneal tear film samples from each group were determined via gelatin zymography for comparison. RESULTS: The proteolytic processes in the ulcerated eyes decreased as corneal healing progressed. On the day of admission, concentrations of latent and active forms of MMP-2 and MMP-9 in ulcerated eyes were significantly higher than values in the contralateral unaffected eyes in dogs with P aeruginosa-associated keratitis; concentrations of latent MMP-2 and MMP-9 were also greater than control group values. Concentrations of latent and active forms of MMP-2 and MMP-9 in the healed eyes of dogs with P aeruginosa-associated keratitis were significantly lower than concentrations in the ulcerated eyes on the day of admission. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that reduction of precorneal tear film concentrations of MMPs by use of proteinase inhibitors may be effective in the treatment of dogs with P aeruginosa-associated keratitis.     

Evaluation of tear film proteinases in horses with ulcerative keratitis

Ulcerative keratitis is a common and potentially blinding ocular disease of horses, capable of progressing to corneal perforation in as little as 24 h. This rapid stromal degeneration is mediated in part by exogenous and endogenous proteinases. We measured and compared the concentrations of two matrix metalloproteinases (MMP-2 and MMP-9) and a serine proteinase (neutrophil elastase) present in the precorneal tear film of normal horses and horses with rapidly progressing ulcerative keratitis. Precorneal tear film samples were collected from 23 ulcerated and 21 unaffected eyes of 23 horses with unilateral ulcerative keratitis, and from 33 normal eyes of 17 control horses. MMP-2, MMP-9, and neutrophil elastase were identified by casein and gelatin zymography and quantified by computerized image analysis. Median MMP-9 levels were significantly higher in the precorneal tear film of young control horses vs. older control horses ( P  = 0.005). Median MMP-2, MMP-9, and neutrophil elastase levels were significantly higher in the precorneal tear film of ulcerated eyes when compared to age-matched normal controls ( P  = 0.004, P  = 0.001, and P  = 0.012, respectively). Median MMP-2 levels were also significantly higher in the precorneal tear film of contralateral eyes of affected horses when compared to age-matched normal controls ( P  = 0.004). No significant differences in median proteinase levels were detected between 'sterile' ulcers and those from which bacteria or mixed infections (bacteria and fungi) were isolated. However, median MMP-2 and neutrophil elastase levels were significantly higher in the precorneal tear film of eyes with 'sterile' ulcers when compared with ulcerated eyes from which fungi were isolated ( P < 0.05). The results of this study support the use of topical antiproteinase therapy which targets both MMPs and serine proteinases in progressive equine ulcerative keratitis.     

Proteinases of the cornea and preocular tear film

Maintenance and repair of corneal stromal extracellular matrix (ECM) requires a tightly coordinated balance of ECM synthesis, degradation and remodeling in which proteolytic enzymes (proteinases) perform important functions. There are natural proteinase inhibitors present in preocular tear film (PTF) and cornea simultaneously with proteinases that prevent excessive degradation of normal healthy tissue. Disorders occur when there is an imbalance between proteinases and proteinase inhibitors in favor of the proteinases, causing pathologic degradation of stromal collagen and proteoglycans in the cornea. Two matrix metalloproteinases (MMPs), MMP-2 and MMP-9, are of major importance in terms of remodeling and degradation of the corneal stromal collagen. Immunohistochemical studies have shown different origins of MMP-2 and -9. MMP-2 is synthesized by corneal keratocytes and performs a surveillance function in the normal cornea, becoming locally activated to degrade collagen molecules that occasionally become damaged. Alternatively, MMP-9 may be produced by epithelial cells and polymorphonuclear neutrophils following corneal wounding. Because the cornea is in close contact with the preocular tear film (PTF), proteinases have been evaluated in the PTF. In damaged corneas, total proteolytic activity in the tear fluid was found to be significantly increased compared to normal eyes and contralateral eyes. Studies analyzing the proteolytic activity in serial PTF samples during corneal healing led to the following conclusions: ulcerative keratitis in animals is associated with initially high levels of tear film proteolytic activity, which decrease as ulcers heal; proteinase levels in melting ulcers remain elevated leading to rapid progression of the ulcers. The success of medical and surgical treatment of the corneal ulcers is reflected by the proteolytic activity in tears. In animals, successful treatment leads to a rapid reduction in tear film proteolytic activity that corresponds with the improvement in the clinical signs of corneal ulceration. The in vitro effects of various compounds on proteolytic activity in the tear fluid of animals with ulcerative keratitis have been evaluated and their important inhibitory effects have been confirmed. Because these various compounds utilize different mechanisms to inhibit various families of proteinases, a combination of these proteinase inhibitors may be beneficial.     

Evaluation of various compounds to inhibit activity of matrix metalloproteinases in the tear film of horses with ulcerative keratitis

OBJECTIVE: To examine in vitro effects of various antiproteolytic compounds on activity of matrix metalloproteinase (MMP)-2 and -9 in the tear film of horses with active corneal ulcers. SAMPLE POPULATION: Samples of tear film obtained from the eyes of 34 horses with active ulcerative keratitis. PROCEDURE: Horses were sedated, and tear samples were collected from the lower fornix of 34 ulcerated eyes by use of capillary tubes. The protease inhibitors 0.2% EDTA, 0.1% doxycycline, 10% N-acetylcysteine (NAC), 0.1% solution of a modified dipeptide that contains hydroxamic acid (ie, ilomostat), 0.1% alpha1-proteinase inhibitor (PI), 0.5% alpha1-PI, and 100% fresh equine serum (ES) were used to treat pooled samples. Amount of latent and active MMP-2 and -9 was measured by optical density scanning of gelatin zymograms of treated and untreated tear samples. RESULTS: Pooled tear samples obtained from ulcerated eyes contained the latent and active forms of MMP-2 and -9. Compared with MMP activity in untreated samples, total MMP activity (sum of all bands detected) observed on the gelatin zymogram gels was reduced by 99.4% by EDTA, 96.3% by doxycycline, 98.8% by NAC, 98.9% by ilomostat, 52.4% by 0.1% alpha1-PI, 93.6% by 0.5% alpha1-PI, and 90.0% by ES. CONCLUSIONS AND CLINICAL RELEVANCE: We documented that EDTA, doxycycline, NAC, ilomostat, alpha1PI, and ES inhibited MMP activity in vitro. Because these compounds use different mechanisms to inhibit various families of proteases in the tear film of horses, a combination of these protease inhibitors may be beneficial for treatment of corneal ulcers in horses.     

Cochet-Bonnet aesthesiometer-determined corneal sensitivity in neonatal foals and adult horses

Corneal touch threshold (CTT) was measured in sick neonatal foals, healthy foals, and healthy adult horses with a Cochet-Bonnet aesthesiometer. The mean overall CTT for the adult horses, sick foals, and healthy foals was 4.82 +/- 0.87 cm, 3.21 +/- 0.24 cm, and 5.01 +/- 0.61 cm, respectively. The central cornea of adult horses was more sensitive than the limbal cornea. Corneal sensitivity was significantly reduced in sick neonatal foals compared to adults. The mean Schirmer I tear test values were significantly lower in foals than adults, and were 14.2 +/- 1.0 mm, 12.8 +/- 2.4 mm, and 18.3 +/- 2.1 mm wetting in sick neonatal foals, normal neonatal foals, and adult horses, respectively. Reduced corneal sensation and lower tear production may be associated with ulcerative keratitis and slow corneal healing in some foals.     

Feline corneal disease

The cornea is naturally transparent. Anything that interferes with the cornea's stromal architecture, contributes to blood vessel migration, increases corneal pigmentation, or predisposes to corneal edema, disrupts the corneas transparency and indicates corneal disease. The color, location, and shape and pattern of a corneal lesion can help in determining the underlying cause for the disease. Corneal disease is typically divided into congenital or acquired disorders. Congenital disorders, such as corneal dermoids are rare in cats, whereas acquired corneal disease associated with nonulcerative or ulcerative keratitis is common. Primary ocular disease, such as tear film instability, adenexal disease (medial canthal entropion, lagophthalmus, eyelid agenesis), and herpes keratitis are associated with the majority of acquired corneal disease in cats. Proliferative/eosinophilic keratitis, acute bullous keratopathy, and Florida keratopathy are common feline nonulcerative disorders. Nonprogressive ulcerative disease in cats, such as chronic corneal epithelial defects and corneal sequestration are more common than progressive corneal ulcerations.     

Doxycycline levels in preocular tear film of horses following oral administration

Objective To determine the concentration of doxycycline in preocular tear film following oral administration in horses as a possible therapeutic modality for infectious and keratomalacic equine keratitis. Procedure Eight broodmares without ocular disease from a Thoroughbred breeding facility were included in this study. Each mare received 20 mg/kg of doxycycline by mouth once daily in the morning for five consecutive days. Tears were collected 1 h after doxycycline administration starting on day one of administration and continuing for 10 consecutive days. Doxycycline levels in the tears were measured using liquid chromatography with tandem mass spectrometric detection (LC-MS/MS). Results Doxycycline was present in the tears of each mare at low µg/mL levels with the highest concentration appearing on the third to fifth days (8.21–9.83 µg/mL). Doxycycline levels had fallen below quantifiable ranges by day 10. No systemic side-effects were noted in any of the horses included in this study. Conclusions Oral doxycycline is present in preocular tear film of normal horses with noninflamed eyes and may be useful as treatment in equine ulcerative keratomalacia. The oral dose listed was tolerated well by the horses in this study. The drug levels attained at 20 mg/kg once daily orally of doxycycline may aid in the treatment of corneal ulceration in horses, but further study is warranted.     

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.     

Matrix metalloproteinase inhibition in corneal ulceration.

The primary objective of current treatment strategies for infectious keratitis is to sterilize the ulcer as rapidly as possible with topically administered antibiotics. Ulcerative processes can proceed in some cases, despite the absence of microbes, as a result of remaining corneal and tear film MMPs. Combining antibiotic therapy with MMP inhibitors can speed corneal healing, because MMPs play an important role in corneal ulceration and stromal liquefaction. MMPs from the rabbit, horse, and human being are inhibited by metal-binding agents EDTA, NAC, and doxycycline as well as by the serum antiprotease alpha2-macroglobulin. It is not yet certain which proteinase inhibitor has the most favorable therapeutic index for clinical use, although we prefer serum because of its effects on multiple types of proteinases. The MMP inhibitors do have significant therapeutic promise in the treatment of corneal ulceration.     

Efficacy of two chondroitin sulfate ophthalmic solutions in the therapy of spontaneous chronic corneal epithelial defects and ulcerative keratitis associated with bullous keratopathy in dogs

OBJECTIVE: To determine the efficacy of two antimicrobial-chondroitin sulfate ophthalmic solutions in the therapy of spontaneous chronic corneal epithelial defects (SCCED) and ulcerative keratitis associated with bullous keratopathy in dogs. ANIMALS STUDIED: Eighty dogs with SCCED and 14 dogs with ulcerative keratitis associated with bullous keratopathy. PROCEDURE: Following manual debridement of nonadherent epithelium, dogs were treated topically with a chondroitin sulfate ophthalmic solution containing either tobramycin or ciprofloxacin. Patients were re-evaluated at 2-week intervals for 4 weeks. RESULTS: After 2 weeks of treatment, 53.6% of eyes with SCCED and 17.6% of eyes with ulcerative keratitis associated with bullous keratopathy had healed. After 4 weeks of treatment, 81.0% of eyes with SCCED and 23.5% of eyes with ulcerative keratitis associated with bullous keratopathy had healed. There were no statistically significant differences in healing percentages between the tobramycin-chondroitin sulfate solution treatment groups and the ciprofloxacin-chondroitin sulfate solution treatment groups. Two dogs with SCCED, one treated with the tobramycin-chondroitin sulfate solution and the other treated with the ciprofloxacin-chondroitin sulfate solution, developed sterile corneal stromal abscesses during the study. CONCLUSIONS: Topical therapy with an antimicrobial-chondroitin sulfate ophthalmic solution combined with manual debridement of nonadherent epithelium compares favorably with other published medical and surgical therapies for SCCED; however, these compounds are only equivocally more effective than therapy with manual debridement alone. These solutions appear to be ineffective in the treatment of ulcerative keratitis associated with bullous keratopathy. The significance of the two cases of corneal stromal abscessation is unknown at this time and warrants further investigation.     

Corneal stromal sequestration and keratoconjunctivitis sicca in a horse

A 19-year-old Shetland pony presented with unilateral ocular discomfort and abnormal ocular appearance. Keratoconjunctivitis sicca, ulcerative keratitis and brown discoloration of the corneal stroma were identified on ophthalmic examination. The etiology of keratoconjunctivitis sicca was not determined in this case. For practical and financial reasons, the owners requested enucleation of the affected eye. Histopathologic examination revealed extensive loss of corneal epithelium overlying a zone of hypereosinophilic, degenerate, and necrotic corneal stroma. This well-circumscribed region of corneal stromal sequestration was surrounded by stromal vascularization, and an intense inflammatory, predominantly polymorphonuclear, cellular infiltrate. The clinical and histopathologic features of this case were considered remarkably similar to those observed in feline corneal stromal sequestration.     

Schirmer tear test, phenol red thread tear test, eye blink frequency and corneal sensitivity in the guinea pig

OBJECTIVE: To establish reference values for Schirmer tear tests (STT) I and II, phenol red thread (PRT) tear test and eye blink frequency, and to determine corneal sensitivity for normal guinea pigs. ANIMALS STUDIED: One hundred and eight eyes of 54 adult Duncan-Hartley guinea pigs. PROCEDURE: Schirmer tear test (STT) I and then STT II were performed in 36 guinea pigs. PRT and STT I were compared in 18 adult Duncan-Hartley guinea pigs. Corneal sensitivity was determined in 23 guinea pigs by evaluating the corneal touch threshold (CTT) of five different regions using a Cochet-Bonnet esthesiometer. Eye blink frequency was measured in 10 guinea pigs over a period of 20 min and in 17 guinea pigs over a period of 10 min. RESULTS: Mean STT I was 0.36 mm +/- 1.09 mm (wetting/min) and mean STT II was 0.43 mm +/- 1.29 mm (wetting/min). There was no significant difference between mean STT I and mean STT II (P = 0.79). The mean PRT-value was 16 +/- 4.7 mm (wetting/15 s), and the mean STT I-value in the same guinea pigs was 0.6 +/- 1.83 mm (wetting/min). Corneal sensitivity was significantly higher in the center than in the four limbal regions. The mean CTT for central, ventral, nasal, temporal and dorsal regions was 2, 1.7, 1.7, 1.7 and 1.6 cm or 3.7, 5.2, 5.6, 5.7 and 6.4 g/mm(2), respectively. Eye blink frequency was between two to five (mean 3.4 +/- 1.04) blinks per eye over 20 min in guinea pigs in their home environment, while in handheld and restrained guinea pigs eye blink frequency showed a variation between 0 and 17 blinks per eye (mean 3.24 +/- 3.64 blinks per eye) over 10 min. CONCLUSIONS: As there were no significant differences between STT I and STT II results, reflex tear secretion in the guinea pig may not exist. The most likely explanation is a lower corneal sensitivity in the guinea pig than in other species, such as cats, dogs and horses. Because of the small amount of tears, PRT is the preferred test for tear measurement in the guinea pig.     

Qualitative tear film and conjunctival goblet cell assessment of cats with corneal sequestra

OBJECTIVES: To evaluate the tear film qualitatively and conjunctival goblet cell numbers in cats with and without corneal sequestra. ANIMALS STUDIED AND PROCEDURES: This was a prospective evaluation of 11 cats with corneal sequestra and 14 control eyes that were either the contralateral normal eye when the sequestrum was unilateral or from control cats of similar age with no ocular disease. All cats in this study were examined by a veterinary ophthalmologist. The ophthalmic examinations included a neuro-ophthalmic evaluation, Schirmer tear tests, fluorescein staining, tear film break-up times, applanation tonometry, biomicroscopy, and indirect ophthalmoscopy. The palpebral conjunctiva at the dorsal nasal, ventral nasal, dorsal temporal and ventral temporal fornices were biopsied after topical anesthetic was applied to the cornea and conjunctiva. The conjunctival biopsies were fixed in formalin and sectioned routinely and stained with hematoxylin and eosin, and periodic acid-Schiff. These slides were examined by light microscopy by a blinded examiner. Goblet cell numbers were compared to conjunctival basal epithelial cell numbers by region. The goblet cell numbers by region from the eyes with sequestra was statistically compared to those from eyes without sequestra, with a student's paired t-test. Conjunctival swabs were collected from the cats with corneal sequestra and submitted for polymerase chain reaction for Herpes felis, Chlamydia psiitticia, and Mycoplasma felis. The corneal sequestra were removed by surgical keratectomy and fixed and stained routinely, and examined by light microscopy. RESULTS: No neurologic abnormalities were detected in any of the cats. The Schirmer tear tests (eyes with sequestra 14+/-5.1 mm/min; normal eyes 15+/-6.8 mm/min) and intraocular pressures (eyes with sequestra 21+/-6.6; normal eyes 22+/-5.8) were within normal reference ranges for cats. Biomicroscopic examinations revealed varied sizes and depths of brown- and amber-colored corneal sequestra. No abnormalities were noted on indirect ophthalmoscopic examinations. The tear film break-up time was 21 s (+/-12) for the normal eyes (n=14) and 14 s (+/-13) in eyes with corneal sequestra (n=11). The average goblet/epithelial cell ratios by region for the normal eyes and the eyes with sequestra respectively were 0.66, 0.56 for the dorsal nasal fornix, 0.68, 0.57 for the ventral nasal fornix, 0.63, 0.48 for the temporal dorsal fornix, and 0.55, 0.49 for the temporal ventral fornix. There were no significant differences in tear film break-up times and goblet cell numbers in eyes with corneal sequestra and those without sequestra. Three conjunctival swabs from two of 11 cats with sequestra were positive with PCR for Herpes felis virus. These included one cat with bilateral sequestra and one cat with unilateral corneal sequestrum. CONCLUSIONS: The pathogenesis of feline corneal sequestra does not appear to be linked primarily to abnormal goblet cell numbers, qualitative tear film abnormalities, and accelerated tear film break-up time.     

Ulcerative keratitis in thoroughbred racehorses in Japan from 1997 to 2008

Objective To describe the number of cases, etiologies, healing times, and results of microbial culture and susceptibility testing of corneal ulcers in thoroughbred racehorses in Japan. Procedure Retrospective study of the medical records of racehorses belonging to the Japan Racing Association (JRA) from 1997 to 2008. Results A total of 2846 horses suffered ulcerative keratitis. These horses accounted for 90.5% of all the corneal problems and 54.9% of the entire number of horses with ocular diseases. Running in races was the cause in 64.3% of the cases. The mean healing time of the ulcers developed in races was 5.5 ± 9.6 days, which was shorter than that of the ulcers unrelated to racing (8.6 ± 11.7 days). In horses presented for examination by the next day after the race, healing was even more rapid (4.1 ± 7.5 days) than in horses presented later (12.4 ± 14.7 days). Microbial culture was performed on 74 samples of 58 horses. Forty‐four bacterial and four fungal organisms were isolated from 35 samples of 29 horses. Pseudomonas aeruginosa (10) and Streptococcus equi subspecies zooepidemicus (5) were the bacteria most frequently isolated. There was no tendency of increasing antibiotic resistance of these organisms. Conclusions Ulcerative keratitis is the most frequent corneal and ocular disease of the racehorse in Japan. Careful observation of the eyes after racing is necessary as early diagnosis and treatment of corneal ulcers speeds healing. Many antibiotic have efficacy against the bacterial isolates, however, potent antibiotics should be reserved for the most severe corneal ulcers.     

Prevalence of equine ulcerative keratomycosis in Colorado and association of environmental factors: A retrospective and descriptive study (2002–2017)

Equine ulcerative keratomycosis is relatively commonly presented to teaching hospitals in North America, with a prevalence from 24 to 86%. Horses in Colorado may have lower risk due to the dry mountain climate, but data are lacking. This study aimed to determine the prevalence of ulcerative keratomycosis amongst horses with ulcerative keratitis presented to Colorado State University Veterinary Teaching Hospital (CSU-VTH) ophthalmology service and to evaluate environmental factors (season, temperature, humidity, wind speed and elevation) associated with ulcerative keratomycosis in Colorado. A database search identified horses with ulcerative keratitis presented to the ophthalmology service at CSU-VTH from January 2002 to August 2017. Sixty-one horses met the inclusion criteria of a corneal cytology and/or culture or histopathology at the time of diagnosis; cases lacking that were excluded. Environmental factors at the boarding sites, including season, temperature, humidity, wind speed and elevation and clinical outcomes for fungal cases, were recorded. Prevalence of fungal infection amongst equine ulcerative keratitis was 16.4% (10/61), suggesting that CSU-VTH has considerably fewer ulcerative keratomycosis cases than other veterinary teaching hospitals in North America. Spring (50%, 5/10) and fall (40%, 4/10) had the highest prevalence of fungal ulcerative keratitis in Colorado. Only one case was reported in summer (10%, 1/10); no horses were positive (0%, 0/10) in winter. Only wind speed seemed to influence the development of ulcerative keratomycosis with higher wind speeds associated with greater rates of fungal involvement (P = 0.047). Other environmental factors did not show a detectable association (all P-values >0.05). Outcomes were variable. It was concluded that horses from the area of CSU-VTH appear to be at lower risk for ulcerative keratomycosis than from the areas around other North American veterinary teaching hospitals that have reported data. Most horses with keratomycosis in this area present in the spring and fall.     

Effects of cyclophotocoagulation with a neodymium:yttrium-aluminum-garnet laser on corneal sensitivity,intraocular pressure,aqueous tear production,and corneal nerve morphology in eyes of dogs

OBJECTIVE: To determine effects of cyclophotocoagulation via administration of 100 J with a neodymium:yttrium aluminum garnet (Nd:YAG) laser on corneal touch threshold (CTT), intraocular pressure (IOP), aqueous tear production, and corneal nerve morphology in eyes of dogs. ANIMALS: 15 dogs. PROCEDURE: Noncontact Nd:YAG laser was transsclerally applied (10 applications; 25 W for 0.1 seconds for each application to each of 4 quadrants) to the ciliary body of the left eye of 15 dogs; the right eye was the control eye. Corneal integrity, CTT, tear production as measured by the Schirmer tear test (STT), and IOP were evaluated for 14 days following laser treatment. On day 14, dogs were euthanatized, eyes harvested, and corneas stained with gold chloride. Major nerve bundles were analyzed by use of a drawing tube attached to a light microscope, and maximum diameters were measured by use of image analysis software. RESULTS: All laser-treated eyes had significantly higher CTT values, compared with control eyes. Six of 15 laser-treated eyes developed ulcerative keratitis. On most days, IOP was significantly lower in laser-treated eyes in both morning and evening. Laser-treated eyes had a significant decrease of approximately 1 nerve bundle/corneal quadrant. Values for STT or nerve bundle diameters did not differ significantly. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of 100 J with a Nd:YAG laser effectively reduced IOP while increasing CTT and caused a significant decrease in number, but not diameter, of major corneal nerve bundles. Nerve damage and corneal hypoesthesia are etiologic factors in ulcerative keratitis following Nd:YAG cyclophotocoagulation.     

The search for causes of nonhealing or recurrent ulcerative keratitis in horses

Chronic or recurrent superficial ulcerative keratitis in horses is common and the causes of these lesions must always be searched for and identified by the clinician. Without appropriate elimination of the underlying cause, the ulcerative keratitis may not resolve, resulting in corneal scarring, decreased vision and possibly loss of the eye. Common causes of chronic or recurrent superficial ulcerative keratitis, such as infectious keratitis, should always be ruled out; however, other possible causes such as single or multiple distichiasis, should be considered.     

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.     

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.