In vitro susceptibility patterns of fungi associated with keratomycosis in horses of the northeastern United States: 68 cases (1987-2006)

OBJECTIVE: To determine in vitro susceptibility patterns of fungi associated with keratomycosis in horses in the northeastern United States and compare those patterns with results of studies from other geographic regions. DESIGN: Retrospective case series. ANIMALS: 68 horses with keratomycosis. PROCEDURES: Medical records of horses with a clinical diagnosis of keratomycosis, positive results of corneal fungal cultures, and susceptibility data were reviewed from the years 1987 to 2006. Fungal identification and in vitro antifungal susceptibility test results were recorded. The percentage of susceptible isolates was compared among antifungals for all isolates together and for the most common genera individually. RESULTS: 74 fungal isolates from 68 horses that met inclusion criteria were identified. Aspergillus, Candida, and Fusarium spp were the most frequent isolates. Grouped isolates had the highest percentage of susceptibility to nystatin (87.7%), natamycin (87.5%), and clotrimazole (80.6%). Grouped isolates had the lowest percentage of susceptibility to fluconazole (15.8%) and miconazole (27.5%). Aspergillus spp (> or = 81.0%) were most susceptible to nystatin, clotrimazole, itraconazole, and natamycin. Candida spp (100%) were most susceptible to ketaconazole, natamycin, and nystatin. Fusarium spp (100%) were only consistently susceptible to natamycin. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of in vitro susceptibility testing, nystatin, natamycin, or clotrimazole is recommended for initial topical treatment of keratomycosis in horses from the northeastern United States. Contrary to results of studies of ocular fungal isolates of horses from other regions, Candida spp were identified more frequently and miconazole had lower in vitro efficacy in the present study.     

In vitro fungistatic and fungicidal activities of silver sulfadiazine and natamycin on pathogenic fungi isolated from horses with keratomycosis

OBJECTIVE: To evaluate the in vitro antifungal properties of silver sulfadiazine (SSD) and natamycin against filamentous fungi isolated from eyes of horses with keratomycosis. SAMPLE POPULATION: Filamentous fungal isolates obtained from eyes of keratomycosis-affected horses. PROCEDURES: Fungal culture of ocular samples yielded 6 Fusarium spp; 7 Aspergillus spp; and 1 isolate each of Curvularia, Scopulariopsis, Penicillium, and Chrysosporium. For each fungal isolate, minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of SSD and natamycin were determined. RESULTS: For all 17 fungal isolates, SSD MIC distribution ranged from < or = 1 to > 64 microg/mL; MIC50 and MIC90 (MICs at which 50% and 90% of organisms were inhibited) were 4 and 32 microg/mL, respectively. The SSD MFC distribution for all isolates was < or = 1 to > 64 microg/mL; MFC50 and MFC90 (MFCs at which 50% and 90% of organisms were killed) were 8 and > 64 microg/mL, respectively. For all fungal isolates, natamycin MIC distribution ranged from 256 to > 1,000 microg/mL; MIC50 and MIC90 were 512 and > 1,000 microg/mL, respectively. The natamycin MFC distribution for all isolates ranged from 512 to > 1,000 microg/mL; MFC(50) and MFC(90) were each > 1,000 microg/mL. CONCLUSIONS AND CLINICAL RELEVANCE: These in vitro data suggest that SSD is fungicidal against the fungal isolates that were obtained from eyes of horses with keratomycosis and that natamycin is fungicidal against some of the isolates at the drug concentrations evaluated. Silver sulfadiazine may be a therapeutic option for equine keratomycosis.     

Susceptibility testing of Aspergillus niger strains isolated from poultry to antifungal drugs--a comparative study of the disk diffusion, broth microdilution (M 38-A) and Etest methods

The aim of this study was to determine the sensitivity of Aspergillus niger strains isolated from birds to available antifungal drugs using different in vitro assays--classical disk diffusion, Etest and broth microdilution NCCLS/CLSI M 38-A. The study material consisted of about 2.000 swabs and samples from different species of birds. A. niger (n=10) was accounted for 6.81% of the total pool of strains isolated. Determinations were made for 13 antifungal drugs using the disk diffusion method. The A. niger exhibited high susceptibility to enilconazole, terbinafine, voriconazole, tioconazole and ketoconazole, low susceptibility to clotrimazole, miconazole and nystatin, and resistance to amphotericin B, itraconazole, pimaricin, fluconazole and 5-fluorocytosine. Minimum inhibitory concentration (MIC) was determined for 9 antifungal drugs using the micromethod of duplicate serial dilutions in a liquid medium. A. niger strains were most susceptible to enilconazole and voriconazole. MIC ranged from 0.0625 to 0.5 microg/ml for enilconazole, with MIC90-0.5 microg/ml and MIC50-0.125 microg/ml. The corresponding values for voriconazole were 0.25-1 microg/ml, 1 microg/ml and 0.5 microg/ml. MIC for amphotericin B and terbinafine ranged from 0.5 to 4 microg/ml, while the values for the remaining drugs were highly varied. MIC was measured by the gradient diffusion method using Etest for 5 antifungal drugs: amphotericin B, fluconazole, itraconazole, ketoconazole and voriconazole. By far the highest susceptibility was obtained in the case of voriconazole, with MIC ranging from 0.0625 to 1 microg/ml. MIC for amphotericin B ranged from 0.25 to 4 microg/ml, for itraconazole and ketoconazole ranging from 0.5 to 16 microg/ml. Methods available for this purpose are not always applicable in field conditions. The present results indicate that the Etest technique, due to its high percentage of agreement with the M 38-A microdilution method, should find application in medical and veterinary practice.     

In vitro efficacy of a buffered chelating solution as an antimicrobial potentiator for antifungal drugs against fungal pathogens obtained from horses with mycotic keratitis

OBJECTIVE: To determine whether a novel third-generation chelating agent (8 mM disodium EDTA dehydrate and 20 mM 2-amino-2-hydroxymethyl-1, 3-propanediol) would act as an antimicrobial potentiator to enhance in vitro activity of antifungal medications against fungal isolates obtained from horses with mycotic keratitis. SAMPLE POPULATION: Fungal isolates (3 Aspergillus isolates, 5 Fusarium isolates, 1 Penicillium isolate, 1 Cladosporium isolate, and 1 Curvularia isolate) obtained from horses with mycotic keratitis and 2 quality-control strains obtained from the American Type Culture Collection (ATCC; Candida albicans ATCC 90028 and Paecilomyces variotii ATCC 36257). PROCEDURE: Minimum inhibitory concentrations (MICs) against fungal isolates for 4 antifungal drugs (miconazole, ketoconazole, itraconazole, and natamycin) were compared with MICs against fungal isolates for the combinations of each of the 4 antifungal drugs and the chelating agent. The Clinical and Laboratory Standards Institute microdilution assay method was performed by use of reference-grade antifungal powders against the fungal isolates and quality-control strains of fungi. RESULTS: Values for the MIC at which the antifungal drugs decreased the growth of an organism by 50% (MIC50) and 90% (MIC90) were decreased for the control strains and ophthalmic fungal isolates by 50% to 100% when the drugs were used in combination with the chelating agent at a concentration of up to 540 microg/mL. CONCLUSIONS AND CLINICAL RELEVANCE: The chelating agent increased in vitro activity of antifungal drugs against common fungal pathogens isolated from eyes of horses with mycotic keratitis.     

In vitro antifungal susceptibility of Malassezia pachydermatis from dogs with and without skin lesions

Canine Malassezia dermatitis is frequently treated with systemic ketoconazole (KTZ) and itraconazole (ITZ). However, no information is available on the antifungal susceptibility to azoles and allilamine of Malassezia pachydermatis isolates from dogs with or without skin lesions. The present study was designed to evaluate the in vitro antifungal susceptibility of M. pachydermatis strains from dogs with or without skin lesions to KTZ, ITZ, miconazole (MICO), fluconazole (FLZ), posaconazole (POS), voriconazole (VOR) and terbinafine (TER) using the Clinical and Laboratory Standards Institute reference Broth Microdilution Method (CLSI M27-A2). The association between the susceptibility to antifungal compounds and the origin of M. pachydermatis, from skin with or without lesions has been also assessed. A total of 62 M. pachydermatis strains from healthy dogs (i.e., Group A=30) or with skin lesions (i.e., Group B=32) were tested. ITZ, KTZ and POS showed the highest activity against M. pachydermatis strains, whereas MICO TER and FLZ the lowest. A higher number of Malassezia resistant strains were registered among isolates from Group B than those from Group A. This study indicates that M. pachydermatis strains were susceptible to ITZ, KTZ, and POS. However, dogs with lesions may harbour strains with low susceptibility to antifungal agents and displaying cross-resistance phenomena to azole. The antifungal therapy in Malassezia infections requires careful appraisal of choice of drugs especially in cases of unresponsiveness to antifungal treatment or recurrent infections.     

The antifungal activity of natamycin toward molds isolated from commercially manufactured poultry feed

The antifungal activity of natamycin, a polyene antifungal compound, was evaluated on molds isolated from commercial poultry feed. The antifungal activity was measured by determination of the minimal inhibitory concentration (MIC) for natamycin on molds growing on semisolid microbiological medium (potato dextrose agar) containing pure natamycin at concentrations ranging from 0 to 200 mg/liter. Natamycin exhibited a high degree of antifungal activity against the 191 isolates of aspergilli used in this study, with average MIC values ranging from 5.08 to 40.1 mg/liter for Aspergillus fumigatus and Aspergillus parasiticus, respectively. Natamycin was also equally effective in inhibiting the growth of nonaflatoxigenic compared with aflatoxigenic isolates of Aspergillus flavus and A. parasiticus. Natamycin was also efficacious against molds other than aspergilli, with MIC values ranging from 2.15 to 5.80 mg/liter for Paecilomyces and Rhizopus spp., respectively. Natamycin exhibited apparent sporicidal activity against spores of toxigenic strains of Fusarium moniliforme and A. parasiticus but not Penicillium rubrum. This sporicidal activity was evident only when spores were exposed to an in vitro concentration of natamycin of 25 mg/liter or higher for a period of time of at least 12 hr. The growth inhibiting activity of natamycin was more pronounced compared with the sporicidal activity.     

Pseudallescheria boydii keratomycosis in a horse

The fungal organism Pseudallescheria boydii was isolated from the cornea of a Quarter Horse with ulcerative keratitis. Despite aggressive hourly medication through a subpalpebral lavage system, with drugs including miconazole and natamycin, the cornea developed a stromal abscess. Orbital exenteration was performed after 3 weeks. The fungal isolate was later determined to be resistant to all 8 antifungal drugs tested. Microscopic examination of the cornea revealed fungal hyphae throughout the corneal stroma and penetrating the Descemet membrane. Pseudallescheria boydii has not been implicated previously as a cause of keratomycosis in horses or in other domestic animals, although cases in human beings have been described.     

Equine keratomycosis in Japan

Objective To describe the incidence, clinical progress, visual outcome, and laboratory findings of equine keratomycosis in Japan. Procedure Retrospective study of the medical records of horses clinically and mycologically diagnosed with keratomycosis at the Equine Hospitals of the Japan Racing Association from 2005 to 2011. Results The diagnosis of keratomycosis was confirmed in eight horses (40.0% of the 20 horses with infectious keratitis from which fungi and/or bacteria were isolated). Fungi recovered from corneal swabs were identified as Aspergillus flavus (4), Aspergillus niger (1), Fusarium solani (1), and Mortierella wolfii (2). All horses were treated medically with topical antifungals, and one horse was also treated surgically. The median of treatment period was 40 days. Two horses were rendered blind in the affected eye and the others retained vision. Conclusions Equine keratomycosis comprises a considerable portion of infectious keratitis in Japan, and the causative fungi that we isolated had been isolated previously from horses with keratomycosis in other regions with the exception of M. wolfii. Culture and cytological examination of corneal lesions should be immediately performed on eyes with signs of keratitis, particularly on those not improving with antibacterial medication, as early initiation of aggressive antifungal treatment tended to result in better outcome and shorter treatment period.     

Susceptibility of fungi isolated from the respiratory tract of falcons to amphotericin B, itraconazole and voriconazole

The minimum inhibitory concentrations (mics) of fungi isolated from the air sacs of falcons before (group 1), and during antifungal treatment with amphotericin B nebulisation and oral itraconazole or voriconazole (group 2), or with itraconazole alone (group 3) or voriconazole alone (group 4) were determined. Before treatment, 95 per cent of the isolates, including Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger and Aspergillus terreus, were susceptible to voriconazole at mics up to 0.38 microg/ml, and all the isolates were susceptible at mics up to 1 microg/ml. Before treatment, 21 per cent of the isolates, including A fumigatus (27.6 per cent), A flavus (16.6 per cent), A niger (100 per cent) and A terreus (23 per cent), were resistant (mic > or =1 microg/ml) to itraconazole; 51 per cent of the isolates, including A fumigatus (31 per cent), A flavus (78 per cent), A niger (14 per cent) and A terreus (77 per cent), had mics of over 1 microg/ml to amphotericin B, and after treatment their mics increased significantly. In contrast, there were no significant differences between the mics of voriconazole and itraconazole for the different Aspergillus species before and during treatment with these antifungal agents.     

The anatomical distribution and antimicrobial susceptibility of yeast species isolated from healthy dogs

The aim of this work was to identify the predominant yeast species present at different anatomical sites in healthy dogs and to determine their in vitro antimicrobial susceptibility using a broth microdilution assay. Samples were collected from the preputial, vaginal, oral and perianal mucosae and the isolates cultured were identified according to their morphological characteristics and biochemical profile. Malassezia pachydermatis was the most commonly isolated yeast, followed by Candida parapsilosis, Candida tropicalis, Candida albicans, Saccharomyces cerevisiae and Rhodotorula spp.Minimum inhibitory concentrations of the azole derivatives ketoconazole, itraconazole and fluconazole against Candida spp. were 0.03–16 μg/mL, 0.06 to >16 μg/mL and 0.5–64 μg/mL, respectively and Candida isolates were sensitive to caspofungin and amphotericin B. Although all isolates of M. pachydermatis were sensitive to itraconazole, fluconazole, ketoconazole and amphotericin B, they were found to be resistant to caspofungin. The study has highlighted that Candida spp., M. pachydermatis, S. cerevisiae and Rhodotorula spp. are part of the normal canine surface microbiota and some of these organisms exhibit in vitro resistance to commonly used antimicrobials.     

Antifungal susceptibility of yeasts isolated from secrection of inflamed mammary glands in cows

Antifungal susceptibility of 150 strains of yeasts isolated from the infected mammary secretion of cows was determined. Their susceptibility to amphoterycin B, nystatin, flucytosine, tioconazole, ketoconazole, miconazole, fluconazole and clotrimazole was evaluated using the disc diffusion method on Yeast Nitrogen Base (YNB) and filter paper discs. The most active antifungal agents in vitro were found to be: tioconazole--96.7%, nystatin--95.4%, amphoterycin--94.0%, and flucytosine--92.7% of susceptible strains; the least active were fluconazole and clotrimazole--39.3% and 60.0% of susceptible strains, respectively. The individual species of yeasts were characterized by varied susceptibility to individual therapeutics.     

In vitro efficacy of lufenuron against filamentous fungi and blood concentrations after PO administration in horses

Lufenuron is a benzoylphenyl urea-derived insecticide that has been recently introduced as a novel treatment for fungal infections in horses. The purposes of this study were to determine (1) the in vitro efficacy of lufenuron against Aspergillus and Fusarium spp. and (2) the ability of lufenuron to reach efficacious blood concentrations after PO administration in horses. Fungal colonies isolated from diseased equine corneas were tested against lufenuron solutions up to 700 microg/mL. Twenty-one adult horses received 1 of 3 PO lufenuron treatment regimens: 5 mg/kg body weight (BW) q24h for 3 days, 20 mg/kg BW q24h for 3 days, or 60 mg/ kg BW q24h for 1 day. Blood samples were collected up to 96 hours after drug administration and analyzed by high-performance liquid chromatography. Statistical analyses of lufenuron blood concentrations were performed by analysis of variance and Fischer's Least Significant Difference test, with statistical significance set at P < .05. Lufenuron showed no effect on the in vitro growth of Aspergillus or Fusarium spp. Lufenuron was detected in the blood of all but 1 horse and showed no adverse effects. The maximum blood lufenuron concentration (83.5 +/- 58.7 microg/L) was lower than the concentrations proven to be ineffective in vitro in this study. Further therapeutic use of lufenuron as an antifungal agent in horses should be based on proven efficacy against specific strains of clinically relevant fungi with pharmacokinetic data demonstrating sufficient lufenuron concentrations in target tissues.     

Prevalence of ocular microorganisms in hospitalized and stabled horses

Microorganisms from normal eyes of hospitalized and stabled horses were identified, and the frequency of isolation was compared between the 2 groups. Using standard techniques, swab specimens from both eyes of 22 hospitalized horses and both eyes of 18 stabled horses were cultured for aerobic bacteria and fungi. Ninety-six aerobic bacteria and 57 fungi were isolated. The predominant bacterial isolates were gram-positive organisms, most of which belonged to the genera Corynebacterium, Bacillus, Staphylococcus, and Streptomyces. Gram-negative organisms comprised less than one-fourth of the bacterial isolates, with the genera Neisseria, Moraxella, and Acinetobacter being the most commonly isolated. Environmental fungi Cladosporium and Alternaria accounted for half of all fungal isolates. In only 5 horses were fungi isolated without accompanying isolation of bacteria. The frequency of isolation of fungi was higher (P less than 0.01) in stabled horses. For bacteria, the frequency of isolation was higher (P less than 0.08) in male horses. Results of susceptibility testing were recorded as the percentage of all isolates susceptible to a given antimicrobic drug. Bacterial isolates were highly susceptible (greater than or equal to 90%) to neomycin, polymixin B, gentamicin, and chloramphenicol. Overall, filamentous fungi had highest susceptibility to natamycin (97%). Miconazole was highly efficacious (100% susceptibility) against Fusarium and Aspergillus.     

In vitro efficacy of an ophthalmic drug combination against corneal pathogens of horses

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 incubation [corrected] 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.     

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.     

Comparison of susceptibility of fungal isolates to lufenuron and nikkomycin Z alone or in combination with itraconazole

OBJECTIVE: To evaluate and compare the in vitro antifungal properties of lufenuron and nikkomycin Z against isolates of Coccidioides immitis and Aspergillus fumigatus when used singly and in combination with the azole antifungal agent itraconazole. SAMPLE POPULATION: 3 clinical isolates of A fumigatus and the Silveira strain of C immitis. PROCEDURE: The fungal isolates were tested in vitro for susceptibility to the single and combination of compounds by use of microtiter-format susceptibility methods. Minimum inhibitory concentration end points were determined visually, and the contents of representative wells were examined microscopically for evidence of morphologic effects on fungi. RESULTS: No evidence of inhibition, either by susceptibility testing or direct microscopic examination of treated cells, was obtained with lufenuron under experimental conditions. In contrast, nikkomycin Z, a known inhibitor of fungal chitin synthesis, had potent activity against C immitis when used singly. A synergistic interaction between nikkomycin Z and itraconazole was found against isolates of both species tested. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of our in vitro data, lufenuron does not appear to possess antifungal properties.     

Antifungal sensitivity testing for equine keratomycosis

We evaluated 31 fungal specimens obtained from equine corneas over a 10-year period, 1973 to 1983. More than half were received in late summer and early autumn, and the number tended to increase in frequency during the 1980s. These isolates included 13 different genera and 20 different species. The prevalent genus was Aspergillus (35%). On the basis of examinations for tube-dilution minimal inhibitory concentrations and minimal fungicidal concentrations of 16 fungal isolates, the imidazole antibiotics such as miconazole and ketoconazole consistently showed the lowest geometric mean titers for filamentous fungi. Because fungal sensitivities varied considerably among different species in vitro sensitivity testing is suggested as an aid in the determination of optimal therapy.     

Keratomycosis in five dogs

Five cases of canine keratomycosis were diagnosed and treated at a private Veterinary Ophthalmology Practice in Melbourne, Australia. Clinical presentations varied between dogs. Predisposing factors were identified in 4 of 5 cases. Diagnostic modalities utilized were corneal cytology and fungal culture. Corneal cytology confirmed the presence of fungal organisms in all five cases. Aspergillus, Scedosporium, and Candida were cultured from three cases, respectively. Specific antifungal treatment included 1% voriconazole solution or 1% itraconazole ointment. Keratectomy and conjunctival grafting surgery was performed in two patients. Resolution of infection and preservation of vision were achieved in 4 of 5 patients.     

Bacterial isolates and antimicrobial susceptibilities in equine bacterial ulcerative keratitis (1993--2004)

REASONS FOR PERFORMING STUDY: Bacterial ulcerative keratitis is a common and often vision-threatening problem in horses. Emerging bacterial resistance to commonly used topical antibiotics has been demonstrated. Previous antibiotic use may alter the antimicrobial susceptibility of bacterial isolates. OBJECTIVES: To document aerobic bacterial isolates and associated bacterial susceptibilities from horses with ulcerative keratitis treated at the University of Tennessee between January 1993 and May 2004 and determine whether prior antibiotic therapy affected antimicrobial susceptibility of the isolates. METHODS: Medical records from horses with ulcerative keratitis and positive aerobic bacterial cultures and antimicrobial susceptibility were evaluated. Clinical history regarding antibiotic therapy prior to culture was documented. RESULTS: Fifty-one aerobic bacterial isolates from 43 horses were identified. Streptococcus equi subspecies zooepidemicus was the most commonly isolated organism, accounting for 33.3% of all isolates, followed by Pseudomonas aeruginosa (11.8%), Staphylococcus spp. (11.8%) and Gram-negative nonfermenting rods (7.8 %). No resistance was noted amongst S. equi ssp. zooepidemicus to cephalothin, chloramphenicol or ciprofloxacin. Only 64 % of S. equi ssp. zooepidemicus isolates were sensitive to bacitracin. No resistance was noted among P. aeruginosa to gentamicin, tobramycin or ciprofloxacin. Antibiotic therapy with neomycin-polymixin B-bacitracin prior to presentation and culture was documented in 11/17 horses in which S. equi ssp. zooepidemicus was isolated and in 4/6 horses in which P. aeruginosa was isolated. Three horses received topical corticosteroids prior to culture, of which 2 had polymicrobial infections. CONCLUSIONS: S. equi ssp. zooepidemicus and P. aeruginosa were the most frequently isolated bacterial organisms in equine ulcerative keratitis. No significant trends in aminoglycoside or fluoroquinolone resistance were noted among these organisms. However, the resistance of S. equi ssp. zooepidemicus to bacitracin with common use of this antibiotic suggests that previous antibiotic therapy probably affects antimicrobial resistance. POTENTIAL RELEVANCE: Therapy prior to culture may play an important role in antimicrobial susceptibility of corneal bacterial isolates. Corticosteroid use may increase the risk of polymicrobial infections of corneal ulcers, leading to a worse prognosis. Although significant fluoroquinolone resistance has not been documented in the veterinary literature, these antimicrobials should be reserved for known infected corneal ulcers and not used for prophylaxis. Empirical antibiotic therapy should not only be guided by clinical signs, but also take into consideration previous antimicrobial and corticosteroid therapy.