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- Author or Editor: Dennis E. Brooks x
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SUMMARY
Objectives
To evaluate in vitro susceptibility to topical antifungal medications, as measured by minimum inhibitory concentration (MIC) and 50% inhibitory concentration (IC50%), of fungal isolates from horses with ulcerative keratomycosis in Florida; to compare results with those of other studies to identify differences in susceptibility patterns among fungi isolated from horses in different geographic regions; and to note indications of fungal resistance to drugs tested in other studies.
Sample Population
Corneal fungal cultures from client-owned horses from Florida with ulcerative keratomycosis (n = 22).
Procedure
Fungal cultures were plated on Emmons modified Sabouraud dextrose agar and mycobiotic agar, examined weekly for growth, and kept for a total of 30 days. In vitro MIC and IC50% of fluconazole, itraconazole, ketoconazole, miconazole, and natamycin were measured for each fungal isolate.
Results
Aspergillus (n = 9; 41%), Fusarium (7; 32%), Penicillium (2; 9%), Cylindrocarpon (1; 4%), Scytalidium (1; 4%), and Torulopsis (1; 4%) spp and an unidentified yeast (1; 4%) were isolated. Fungi were most susceptible to antifungal drugs in the following order: natamycin and miconazole equally, itraconazole, and ketoconazole, although no significant difference was found among drugs. Fungi were significantly less susceptible to fluconazole (P < 0.0001) than to the other 4 drugs.
Conclusions
Initial antifungal therapy with topically applied natamycin, miconazole, itraconazole, or ketoconazole is recommended for ulcerative keratomycosis in horses in the subtropical environment of Florida.
Clinical Relevance
Specific antifungal treatment of horses with ulcerative keratomycosis should be based on history, results of ophthalmic examination, cytologic findings, isolation of the pathogenic fungus, and known prevalence of unique ocular fungi in specific geographic areas. In vitro antifungal susceptibility testing may be most beneficial in aiding documentation of pharmacologic susceptibility patterns of fungi in specific geographic regions. (Am J Vet Res 1998; 59:138–142)
SUMMARY
Oscillatory potentials (op) and electroretinograms (erg) were recorded from clinically normal dogs after 5, 10, 15, 20, 30, 40, 50, and 60 minutes of dark adaptation. At the end of the adaptation period, op were characterized by 5 distinct positive peaks, 01 through 05, with mean latencies of 14.46, 20.24, 27.38, 35.31, and 44.85 ms, respectively, and with mean amplitudes ranging from 7.20 to 34.84 μV. After 60 minutes of dark adaptation, the erg had a mean a-wave latency of 12.03 ms and a mean b-wave amplitude of 109.29 μV. Peaks 03 and 04, which partially mask the summit of the b-wave, had mean latencies of 28.66 and 36.83 ms, respectively. The mean amplitude of the b-wave measured to the peak of 03 was 240.06 μV and 230.73 (μV when measured to peak 04. Changes in the op during dark adaptation consisted of significant (P < 0.05) increases in the latencies of 01, 02, and 03, and significant increases in the amplitudes of 01, 03, 04, and 05. Concurrent erg changes consisted of significant increases in the amplitudes of the a-wave and b-wave measured from 03 and 04, and significant increases in the latencies of peaks 03 and 04 on the b-wave.
Abstract
Objective—To determine density of corneal endothelial cells and corneal thickness in eyes of euthanatized horses.
Sample Population—52 normal eyes from 26 horses.
Procedure—Eyes were enucleated after horses were euthanatized. Eyes were examined to determine that they did not have visible ocular defects. Noncontact specular microscopy was used to determine density of corneal endothelial cells. Corneal thickness was measured, using ultrasonic pachymetry or specular microscopy.
Results—Mean density of corneal endothelial cells was 3,155 cells/mm2. Cell density decreased with age, but sex did not affect cell density. Values did not differ significantly between right and left eyes from the same horse. Cell density of the ventral quadrant was significantly less than cell density of the medial and temporal quadrants. Mean corneal thickness was 893 µm. Sex or age did not affect corneal thickness. Dorsal and ventral quadrants were significantly thicker than the medial and temporal quadrants and central portion of the cornea. We did not detect a correlation between corneal thickness and density of endothelial cells in normal eyes of horses.
Conclusion and Clinical Relevance—Density of corneal endothelial cells decreases with age, but corneal thickness is not affected by age or sex in normal eyes of horses. The technique described here may be useful for determining density of endothelial cells in the cornea of enucleated eyes. This is clinically relevant for analyzing corneal donor tissue prior to harvest and use for corneal transplantation. (Am J Vet Res 2001;62:(479–482)
SUMMARY
Mean (± SD) value for results of the Schirmer tear test without use of topical anesthesia on 142 rabbit eyes was 5.30 (± 2.96) mm/min; range was 15 mm/min, with low value of 0 and high value of 15 mm/min. Two SD from the mean provided a normal clinical range of 0 to 11.2 mm/min for inclusion of 95% of the population. Significant differences were not evident between eye (right vs left) or gender or among breeds; interaction among eye, gender, and breed also was not observed. Although mean values for 2 breeds were significantly different from values in the other 10 breeds and from each other, they were still within the proposed normal range of 2 SD. The Schirmer tear test without topical anesthesia may be useful in rabbits for evaluation of increased values correlated with ocular irritation, rather than for determination of decreased values associated with keratoconjunctivitis sicca.
Abstract
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% α1-proteinase inhibitor (PI), 0.5% α1-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% α1-PI, 93.6% by 0.5% α1-PI, and 90.0% by ES.
Conclusions and Clinical Relevance—We documented that EDTA, doxycycline, NAC, ilomostat, α1- PI, 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. (Am J Vet Res 2003;64:1081–1087)
Abstract
Objective—To evaluate the in vitro efficacy of an ophthalmic drug combination against common corneal pathogens of horses.
Sample Population—Representative isolates of 3 bacterial and 2 fungal corneal pathogens of horses.
Procedures—Pathogens were subjected to minimum inhibitory concentration (MIC) testing of a drug combination that consisted of equal volumes of natamycin 3.33%, tobramycin 0.3%, cefazolin 5.5%, and equine serum. Proteinase inhibitory activity of the drug combination was assessed by use of a fluorescence microplate assay with gelatin and collagen I as substrates. The MICs of the drug combination were compared with those for each of the component medications and antiproteinase activity of the drug combination was compared with that of serum by use of paired t tests and a 2-way ANOVA, respectively.
Results—The drug combination was at least as effective as each medication separately for inhibiting microbial growth of all pathogens tested and was significantly more effective against B-hemolytic Streptococcus spp, Aspergillus spp, and Fusarium spp than the relevant medications separately. Serum and the drug combination both had significant antigelatinase activity, and serum had significant anticollagenase activity. Antiproteinase activity of serum was a concentration-dependent event, which enabled serum to achieve significantly greater activity than the drug combination after 3.5 and 4 hours of intubation for the gelatin and collagen I assays, respectively.
Conclusions and Clinical Relevance—Drug combinations have the attractive potential of minimizing the time, stress, and fatigue associated with topical treatment regimens consisting of multiple drugs used separately for horses with keratitis.
Abstract
Objective—To determine regional and zonal variation in sulfation patterns of chondroitin sulfate in normal equine corneal stroma.
Sample Population—22 normal eyes from 11 horses.
Procedure—Corneas were collected within 24 hours of death from equine necropsy specimens. After papain-chondroitinase digestion of corneal tissue, disaccharides ΔDi4S and ΔDi6S were quantified by use of capillary zone electrophoresis in the superficial, middle, and deep zones of central and peripheral regions of the cornea.
Results—For the 2 regions combined,ΔDi6S/ΔDi4S values were significantly lower in the deep and middle zones, compared with that of the superficial zone. In the central region, deep and middle zones had significantly lower ΔDi6S/ΔDi4S values than the superficial zone did. In the peripheral region, the deep zone had significantly lower ΔDi6S/ΔDi4S values, compared with superficial and middle zones. In the deep zone, the peripheral region had significantly lower ΔDi6S/ΔDi4S values than the central region did.
Conclusion and Clinical Relevance—Distribution of ΔDi6S/ΔDi4S values follows a gradient across the healthy equine cornea, being smallest in the deep and middle zones of the central region and the deep zone of the peripheral region. Regional and zonal differences in the distribution of stromal ΔDi6S and ΔDi4S may influence the role of glycosaminoglycans in health, disease, and wound repair of the equine cornea. (Am J Vet Res 2002;63:143–147)
Abstract
Objective—To examine postoperative ocular hypertension (POH) and other variables as predictors of the risk of developing glaucoma after cataract surgery in dogs.
Design—Retrospective study.
Animals—220 dogs that had cataract surgery.
Procedure—Medical records of 220 dogs (346 eyes) that had extracapsular cataract removal or phacoemulsification of cataracts were reviewed. With respect to glaucoma development, 8 variables were analyzed, which included development of POH, breed, sex, age at time of surgery, eye (right vs left), phacoemulsification time, intraocular lens (IOL) placement (yes or no), and stage of cataract development. Eyes developed glaucoma within 6 or 12 months of surgery or did not have signs of glaucoma at least 6 or 12 months after cataract surgery.
Results—Of 346 eyes, 58 (16.8%) developed glaucoma after surgery. At 6 months, 32 of 206 (15.5%) eyes examined had glaucoma; at 12 months, 44 of 153 (28.8%) eyes examined had glaucoma. Median follow-up time was 5.8 months (range, 0.1 to 48 months). Mixed-breed dogs were at a significantly lower risk for glaucoma, compared with other breeds. Eyes with IOL placement were at a significantly lower risk for glaucoma, compared with eyes without IOL placement. Eyes with hypermature cataracts were at a significantly higher risk for glaucoma, compared with eyes with mature or immature cataracts.
Conclusions and Clinical Relevance—Multiple factors appear to contribute to the onset of glaucoma in dogs after cataract surgery. Complications prohibiting IOL placement during cataract surgery may lead to a high risk of glaucoma development. (J Am Vet Med Assoc 2000;216:1780–1786)
Abstract
Objective—To evaluate clinical characteristics and breeds affected with bacterial keratitis and compare patterns of resistance in bacterial isolates over time in dogs.
Design—Retrospective cross-sectional study.
Animals—97 dogs with bacterial keratitis.
Procedure—Dogs with bacterial keratitis were identified from teaching hospital medical records at the Universities of Tennessee and Florida during the years 1993 to 2003. Data were collected pertaining to breed, Schirmer tear test results, treatments administered at the time of initial examination, bacterial species isolated, and resistance to selected antimicrobials.
Results—66% of the dogs were brachycephalic, 54% had tear production < 15 mm/min, and 29% were receiving a corticosteroid at the time of initial examination. The most common bacteria isolated were Staphylococcus intermedius (29%), β-hemolytic Streptococcus spp (17%), and Pseudomonas aeruginosa (21%). Staphylococcus intermedius isolates had limited resistance to certain antimicrobials. More than 80% of β-hemolytic Streptococcus spp isolates were resistant to neomycin, polymyxin B, and tobramycin. Isolates of P aeruginosa were susceptible to tobramycin and gentamicin and had limited resistance to ciprofloxacin and enrofloxacin. Among bacterial species isolated, there was no evidence of development of antimicrobial resistance over time.
Conclusions and Clinical Relevance—Data suggested that administration of ciprofloxacin or a combination of a first-generation cephalosporin and tobramycin may be used in the treatment of bacterial keratitis while awaiting results of bacterial culture and susceptibility testing. Evidence suggests that current methods of medical management of bacterial keratitis are not associated with increased antimicrobial resistance.