Pharmacokinetics of topically applied ciprofloxacin in equine tears

OBJECTIVE: To evaluate the pharmacokinetics of topically applied ciprofloxacin 0.3% ophthalmic solution in tears of healthy horses. ANIMAL STUDIED: Twenty healthy, adult, mixed-breed horses. PROCEDURES: Twenty study horses were confirmed free of ophthalmic disease by complete ophthalmic examination. Seventy microliters of 0.3% ciprofloxacin (Ciloxan) was placed in the ventral cul-de-sac of each eye using a microliter syringe and 19-g cannula. Population kinetics were carried out by sampling the tear film from the lower cul-de-sac of each eye with tear test strips at 5, 10, 15 and 30 min and 1, 2, 4 and 6 h post administration for a total of five samples at each time-point. Sample collection time was 15 s. Concentrations of ciprofloxacin were determined using high performance liquid chromatography. RESULTS: Mean (+/-SD) of the Schirmer tear test results from all eyes was 23.4 +/- 4.8 mm wetting in 1 min. Mean concentration of ciprofloxacin in the tears at 5 min post administration was 498.4 +/- 266.8 microg/g. Mean concentration rapidly declined and began to plateau at 30 min. The mean tear concentrations of ciprofloxacin at 30 min and at 1, 2, 4 and 6 h were 66.6 +/- 56.0, 60.25 +/- 55.7, 42.25 +/- 30.9, 36.25 +/- 32.0, and 45.5 +/- 46.5 microg/g, respectively. CONCLUSIONS: The pharmacokinetics of ciprofloxacin in normal horses are similar to those in rabbits and humans. Topical application of ciprofloxacin resulted in a mean tear concentration of ciprofloxacin that remained above the MIC(90) levels for most pathogenic bacteria for 6 h post administration.     

Determination of tear break-up time reference values and ocular tolerance of tetracaine hydrochloride eyedops in healthy horses

Reasons for performing study: Tetracaine hydrochloride (THCl) has been reported to cause irritation in dogs. In man, some topical anaesthetics have been shown to disrupt the tear film. Tear break‐up time (TBUT) is a useful test allowing an assessment of the quality of the precorneal tear film. Only one TBUT value has been reported in horses with no information on the technique used. Objectives: To provide a method for performing the TBUT in horses and to report any side effects of a single application of THCl in clinically normal horses, particularly on the stability of the tear film. Methods: In Study 1, one drop of 0.5 or 1% THCl was applied to one eye of 20 horses divided in 2 groups. Treated eyes were assessed for the development of side effects 2.5 and 5 min after treatment. In Study 2, the TBUT was measured in both eyes of 2 groups of 10 horses, before and 2.5 and 5 min after, instillation of one drop of either 0.5 or 1% THCl. Results: No animals developed any ocular side effect after instillation. Basal TBUT was 8.3 ± 1.3 s. TBUT decreased from baseline 5 and 2.5 min after application of one drop of 0.5% THCl and one drop of 1% THCl, respectively. Conclusions: A technique to measure the TBUT in healthy horses is described and normal range values that could be used as a reference were obtained. Potential relevance: THCl is well tolerated in horses but lowers the TBUT.     

Evaluation of concentration of voriconazole in aqueous humor after topical and oral administration in horses

OBJECTIVE: To determine penetration of topically and orally administered voriconazole into ocular tissues and evaluate concentrations of the drug in blood and signs of toxicosis after topical application in horses. ANIMALS: 11 healthy adult horses. PROCEDURE: Each eye in 6 horses was treated with a single concentration (0.5%, 1.0%, or 3.0%) of a topically administered voriconazole solution every 4 hours for 7 doses. Anterior chamber paracentesis was performed and plasma samples were collected after application of the final dose. Voriconazole concentrations in aqueous humor (AH) and plasma were measured via high-performance liquid chromatography. Five horses received a single orally administered dose of voriconazole (4 mg/kg); anterior chamber paracentesis was performed, and voriconazole concentrations in AH were measured. RESULTS: Mean +/- SD voriconazole concentrations in AH after topical administration of 0.5%, 1.0%, and 3.0% solutions (n = 4 eyes for each concentration) were 1.43 +/- 0.37 microg/mL, 2.35 +/- 0.78 microg/mL, and 2.40 +/- 0.29 microg/mL, respectively. The 1.0% and 3.0% solutions resulted in significantly higher AH concentrations than the 0.5% solution, and only the 3.0% solution induced signs of ocular toxicosis. Voriconazole was detected in the plasma for 1 hour after the final topically administered dose of all solutions. Mean +/- SD voriconazole concentration in AH after a single orally administered dose was 0.86 +/- 0.22 microg/mL. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that voriconazole effectively penetrated the cornea in clinically normal eyes and reached detectable concentrations in the AH after topical administration. The drug also penetrated noninflamed equine eyes after oral administration. Low plasma concentrations of voriconazole were detected after topical administration.     

Effect of topical 1% tropicamide on Schirmer tear test results in clinically normal horses

Objective  To observe the effect of topical 1% tropicamide on equine tear production as measured by Schirmer I tear test.
Materials and methods  Fourteen adult horses received one drop of 1% tropicamide ophthalmic solution in one eye and the opposite eye served as the control. The tear production in both eyes was tested at 1, 2, 4, 6, and 24 h after 1% tropicamide administration.
Results  Measurements made 1 h after treatment revealed a significant reduction in Schirmer tear test values in tropicamide treated eyes ( P  = 0.002). The observed decrease in tear production was maintained up to 4 h after treatment ( P  = 0.002). Although tropicamide-induced decrease in STT values was observed in the treated eyes, the contralateral eyes did not show significant changes in Schirmer tear test results.
Conclusion  Single dose of topical 1% tropicamide resulted in statistically significant reduction in Schirmer tear test values in clinically normal horses.     

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.     

Inferomedial placement of a single-entry subpalpebral lavage tube for treatment of equine eye disease

The objective of this study was to describe method of placement, and frequency and severity of complications associated with a subpalpebral lavage system placed in the medial aspect of the equine inferior eyelid. The inferomedial subpalpebral lavage (ISPL) tube is positioned deep in the medial aspect of the inferior conjunctival fornix so that the footplate lies flat between the lower eyelid and the anterior surface of the nictitans. Retrospective data from the placement of 92 ISPL systems placed in 86 horses during a 31-month period were examined. Tube placement was performed using sedation and regional anesthesia only in 59% of horses. The median duration of tube placement was 19 days (range: 1-61 days). Seventy-one horses were treated for up to 55 days following discharge from hospital with an ISPL tube in place. No complications were reported with 59% of ISPL systems. Non-ocular complications were found in 38% of ISPL systems and included tube displacement from the conjunctival fornix (18%), suture loss requiring resuturing of the system to the horse's head (14%), and damage necessitating replacement of the injection port (6%). Ocular complications were recorded in 3% of horses and were limited to inferior eyelid swelling. Vision was retained in 88% of horses. The ISPL system is easily and safely placed, and well tolerated for extended periods. It appears to be associated with infrequent and minor complications when compared with placement of subpalpebral lavage tubes in the superior eyelid.     

In Vitro Evaluation of the Antibiotic Activity of Combinations of Ophthalmic Drugs Against Common Equine Ocular Pathogens

A subpalpebral lavage catheter is commonly used to administer medications in the treatment of infectious keratitis in horses. Drugs may mix in the subpalpebral lavage catheter, potentially affecting antimicrobial efficacy. The objective of this study was to determine the in vitro antimicrobial efficacy of combinations of commonly used ophthalmic preparations. An agar gel diffusion bioassay was used to determine the antibiotic activity of a number of antimicrobial, antifungal, mydriatic, and antiproteinase drug combinations. Drug mixtures were allowed to sit in a syringe at ambient temperatures and room lighting, for 1 (T1) and 6 hours (T2). The antibacterial efficacy was determined by measuring the zone of inhibition around wells containing the drug combinations after overnight incubation at 37°C and comparing them with the zone of inhibition around the antimicrobial alone. Combinations were tested against Pseudomonas aeruginosa and Streptococcus zooepidemicus. There was no significant decrease in the zone of inhibition, compared with controls, for the majority of combinations tested. Significantly smaller zones of inhibition were identified for the following combinations: (1) tobramycin, cefazolin, natamycin, and serum (T1) and (2) tobramycin, cefazolin, natamycin, serum, and atropine (T1) compared with tobramycin alone; (3) ciprofloxacin, cefazolin, natamycin (T1) compared with ciprofloxacin alone; (4) triple antibiotic ophthalmic solution (TAB) and natamycin (T6); (5) TAB, natamycin, and serum (T6); and (6) TAB, natamycin, serum, and atropine (T6) compared with TAB alone. Admixture does not affect in vitro antibacterial efficacy for the majority of ophthalmic preparations, although efficacy of TAB may be decreased compared with the drug alone.     

Systemic dexamethasone concentration in horses after continued topical treatment with an ophthalmic preparation of dexamethasone.

OBJECTIVE: To determine concentrations of dexamethasone in serum and urine of horses treated repeatedly with a topically administered ophthalmic dexamethasone preparation. ANIMALS: 4 clinically normal horses (2 mares, 2 geldings). PROCEDURE: 0.1% dexamethasone ophthalmic ointment was administered to the left eye of each horse every 5 to 9 hours for 8 consecutive days, yielding an estimated cumulative dexamethasone dose of 6.4 microg/kg of body weight. Serum and urine samples were obtained before the first dexamethasone treatment, on days 4 and 8 of treatment, and 24, 48, and 96 hours after cessation of treatment. To detect small concentrations of dexamethasone, serum and urine samples were analyzed by use of a competitive enzyme immunoassay. RESULTS: During the period of continued topical treatment, serum dexamethasone concentrations increased to between 0.10 and 0.49 ng/ml, then decreased below the limit of detection (0.06 ng/ml) within 24 hours after cessation of treatment. Dexamethasone also was detected in urine samples at concentrations of up to 0.98 ng/ml. CONCLUSIONS: Repeated topical administration of dexamethasone ophthalmic ointment generated low, but detectable glucocorticoid concentrations in serum and urine. CLINICAL RELEVANCE: Because treatment of performance horses with dexamethasone is prohibited for most types of competitions and because enhanced glucocorticoid detection methods may result in positive test results, owners and trainers may wish to reconsider entering horses in competitions during periods of treatment with ophthalmic dexamethasone preparations.     

Effect of lacrimal punctal occlusion on tear production and tear fluorescein dilution in normal dogs

OBJECTIVE: To evaluate effects of lacrimal punctal plugs positioned in either the upper, lower, or combination of upper and lower lacrimal canaliculi on plug retention and tolerance; tear production, as measured by the Schirmer tear test; and the dilution of fluorescein within the tear film in normal dogs. MATERIAL AND METHODS: Lacrimal punctal plugs were positioned in the lower, upper, or combination of lower and upper plugs in six laboratory-quality Beagles under topical anesthesia. Retention of plugs was evaluated daily from 8 to 23 days by visual inspection and slit-lamp biomicroscopy. Schirmer tear tests (STT 1 without topical anesthesia) were performed at 48-h intervals. Dilution of fluorescein was determined at 5- and 45-min post-fluorescein instillations once weekly. RESULTS: Lacrimal punctal plugs of 0.4 and 0.6 mm in diameter were retained for 14 (lower plugs: 100%) and 23 days (75%), and for the upper plugs at 8 days less often (75%), and were infrequently locally nonirritating. Combination of lower and upper plugs seemed to adversely affect retention of either plug. When loss of the plugs occurred, a next larger size plug was necessary suggesting some stretching of the lacrimal canaliculi occurred. Pre- and postplug placement STT results indicated no change with lower and combination lacrimal punctal plugs, but decreased levels following upper lacrimal punctal plugs. Tear fluorescein levels at 5 and 45 min in control eye (no punctum plugs) were 3.39% and 0.14%, respectively. With lower, upper, and the combination of lower and upper lacrimal puncta plugs, tear fluorescein levels at 45 min were higher than the controls (lower: 0.76%; upper: 0.45%, and combination 0.56%). CONCLUSION: Lacrimal punctal silicone plugs are retained for 8-23 days in the lower, upper, and combined lower and upper canaliculi at high rates. Effects on STT levels appear limited. Fluorescein within the tear film persists longer with all different positioned lacrimal punctum plugs than in the control eyes.     

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.     

Subconjunctivally implanted micro-osmotic pumps for continuous ocular treatment in horses.

OBJECTIVE: To evaluate the feasibility of using a subconjunctivally implanted micro-osmotic pump for continuous delivery of medication to the eyes of horses- during a 7-day period. ANIMALS: 4 healthy adult horses. PROCEDURE: With horses restrained in a standing position, micro-osmotic pumps were implanted subconjunctivally in each eye for 7 days. The treatment eye received an atropine-loaded micro-osmotic pump (100 microl of 1.5% atropine), and the contralateral eye received a sterile saline-loaded pump (100 microl of 0.9% NaCl) as a control treatment. Pupil size was measured at 12-hour intervals until values returned to baseline. RESULTS: The micro-osmotic pumps were tolerated and did not migrate or become dislodged. During the 7-day treatment period, pupils were significantly larger in the eyes implanted with atropine-loaded pumps, compared with saline-implanted control eyes. CONCLUSIONS AND CLINICAL RELEVANCE: Micro-osmotic pumps were implanted and removed easily from standing horses and were not associated with complications during the 7-day treatment period. Therefore, subconjunctivally implanted micro-osmotic pumps can potentially be used when treating ophthalmic disease in horses.     

Serum concentrations of lidocaine and its metabolites after prolonged infusion in healthy horses

REASONS FOR PERFORMING STUDY: Continuous-rate infusions (CRI) of lidocaine are often used for prolonged duration but, to date, only limited time/concentration relationships administered as a short term (24 h) CRI have been reported. OBJECTIVE: To determine the time/concentration profile of lidocaine and its active metabolites glycinexylidide (GX) and monoethylglycinexylidide (MEGX) during a 96 h lidocaine infusion. METHODS: Lidocaine was administered to 8 mature healthy horses as a continuous rate infusion (0.05 mg/kg bwt/min) for 96 h. Blood concentrations of lidocaine, GX and MEGX were determined using high performance liquid chromatography during and after discontinuation of the infusion. RESULTS: Serum lidocaine concentrations reached steady state by 3 h and did not accumulate thereafter. Concentrations were above the target therapeutic concentration (980 ng/ml) only at 6 and 48 h, and did not reach the range described as potentially causing toxicity (>1850 ng/ml) at any time. MEGX did not accumulate over time, while the GX accumulated significantly up to 48 h and then remained constant. The serum concentrations of lidocaine, MEGX and GX were below the limit of detection within 24 h of discontinuation of the infusion. None of the horses developed any signs of lidocaine toxicity during the study. CONCLUSIONS: The metabolism of lidocaine was not significantly impaired by prolonged infusion and no adverse effects were observed. Prolonged infusions appear to be safe in normal horses but the accumulation of GX, a potentially toxic active metabolite, is cause for concern.     

Continuous infusion of gentamicin into the tarsocrural joint of horses

OBJECTIVE: To develop a method for continuous infusion of gentamicin into the tarsocrural joint of horses, to determine pharmacokinetics of gentamicin in synovial fluid of the tarsocrural joint during continuous infusion, and to evaluate effects of continuous infusion of gentamicin on characteristics of the synovial fluid. ANIMALS: 12 healthy adult horses. PROCEDURE: An infusion catheter consisting of flow control tubing connected to a balloon infuser was used. Gentamicin solution (100 mg/ml) was infused in the right tarsocrural joint and balanced electrolyte solution was infused in the left tarsocrural joint for 5 days. Synovial fluid and serum gentamicin concentrations were measured by use of a fluorescence polarization immunoassay. RESULTS: 17 of the 24 (71%) infusion catheters initially placed functioned without complications for the entire 5-day infusion period. Median gentamicin concentration in synovial fluid from treated joints during the 5-day infusion period ranged from 2875 to 982 microg/ml. Median serum gentamicin concentration during this period ranged from 2.31 to 2.59 microg/ml. Mean (+/- SD) elimination half-life and total clearance of gentamicin from the synovial fluid were 6.25+/-1.01 hours and 1.52+/-0.96 ml/min, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: An infusion catheter can be used for continuous infusion of gentamicin into the tarsocrural joints of horses for up to 5 days. At a gentamicin dosage of 0.17+/-0.02 mg/kg/h, continuous intra-articular infusion results in synovial fluid gentamicin concentrations greater than 100 times the minimal inhibitory concentration reported for common equine pathogens.     

Increased drug concentration and repeated eye drop administration as strategies to optimize topical drug delivery: A fluorophotometric study in healthy dogs

Objectives

Determine tear film kinetics with different fluorescein concentrations and repeated eye drop administration at various time intervals.

Animals Studied

Six healthy Beagles.

Procedures

Six experiments were conducted on separate days: single eye drop administration (control) or two separate eye drops administered at 30 s, 1, 2, 5, and 10 min intervals. For each experiment, one eye received 0.3% fluorescein solution while the other eye received 1% fluorescein solution, and tear fluid was collected with capillary tubes at 0, 1, 5, 10, 20, 30, 40, 50, 60, 90, 120, and 180 min. Fluorescein concentrations were measured using automated fluorophotometry.

Results

Compared with 0.3% solution, eyes receiving 1% fluorescein solution had significantly higher tear film concentrations (p ≤ .046) and the area-under-the-fluorescein-time curve was twofold greater (p = .005). Compared with control: (i) Tear film concentrations were significantly higher for up to 20 min when repeating administration 30 s to 5 min after the first drop (p ≤ .006); (ii) The highest increase in area-under-the-curve was obtained with 2 and 5 min intervals for 0.3% (+109%–130%) and 1% solutions (+153%–157%); (iii) The highest increase in median precorneal retention time (defined as tear film concentration < 5% from baseline values) was obtained with 5 min intervals for 0.3% (55 min vs. 15 min in control) and 2–5 min intervals for 1% solutions (50 min vs. 25 min in control).

Conclusions

Drug delivery to the ocular surface can be enhanced by using more concentrated formulations and/or by repeating eye drop administration 2–5 min after the first dose.     

Pharmacokinetics and pharmacodynamics of terbutaline in healthy horses

OBJECTIVE: To determine pharmacokinetics of terbutaline in healthy horses and to relate serum terbutaline concentrations with the drug's pharmacodynamic effects. ANIMALS: 6 healthy horses. PROCEDURE: Horses were given terbutaline i.v. (10 microg/kg of body weight) and, 1 week later, p.o. (100 microg/kg). Responses to drug administration (eg, heart rate and serum lactate concentration) were measured. Serum terbutaline concentration was measured by means of gas chromatography with mass spectrometry. Protein binding was determined in vitro. RESULTS: Following i.v. administration, median maximum serum terbutaline concentration and mean residence time were 9.3 ng/ml and 30 minutes, respectively. Bioavailability following oral administration was < 1%. All horses developed sweating, trembling, excitement, and tachycardia during i.v. infusion. The 2 horses with the highest serum terbutaline concentrations developed severe tachycardia and CNS stimulation; 30 minutes after the i.v. infusion was completed, they were hyperventilating and lethargic. Heart rate and serum lactate concentration increased as serum terbutaline concentration increased. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that terbutaline is rapidly cleared from the bloodstream following i.v. administration to horses, suggesting that continuous i.v. infusion would be needed to maintain therapeutic serum concentrations. Oral administration of terbutaline to horses is not practical because of the low bioavailability.     

Use of free conjunctival grafts in horses: ten cases

OBJECTIVE: To assess the effectiveness of free conjunctival grafts in the treatment of horses with a range of keratopathies. DESIGN: A retrospective clinical study of ten client-owned horses treated at Murdoch University Veterinary Hospital from May 1996 to September 2001. PROCEDURE: The suitability of patients for the surgical procedure was assessed using a slit lamp biomicroscope and by direct and indirect ophthalmoscopy. Surgery was performed with the aid of an operating microscope, under general anaesthesia. A subpalpebral ocular lavage catheter was used for administration of topical atropine and antibiotics postoperatively. RESULTS: In all ten horses the affected globe was saved. In nine of the horses vision in the eye was satisfactory 6 months after surgery, and in one horse the eye was blind. Complications included further corneal ulceration or eyelid abscessation and some loss of sutures, although these did not preclude a successful outcome. CONCLUSION: Free conjunctival grafts were successful in treating a range of keratopathies in the horse, and the technique offers a number of advantages over other forms of surgical intervention.     

Pharmacokinetics of voriconazole following intravenous and oral administration and body fluid concentrations of voriconazole following repeated oral administration in horses

OBJECTIVE: To determine the pharmacokinetics of voriconazole following IV and PO administration and assess the distribution of voriconazole into body fluids following repeated PO administration in horses. ANIMALS: 6 clinically normal adult horses. PROCEDURES: All horses received voriconazole (10 mg/kg) IV and PO (2-week interval between treatments). Plasma voriconazole concentrations were determined prior to and at intervals following administration. Subsequently, voriconazole was administered PO (3 mg/kg) twice daily for 10 days to all horses; plasma, synovial fluid, CSF, urine, and preocular tear film concentrations of voriconazole were then assessed. RESULTS: Mean +/- SD volume of distribution at steady state was 1,604.9 +/- 406.4 mL/kg. Systemic bioavailability of voriconazole following PO administration was 95 +/- 19%; the highest plasma concentration of 6.1 +/- 1.4 microg/mL was attained at 0.6 to 2.3 hours. Mean peak plasma concentration was 2.57 microg/mL, and mean trough plasma concentration was 1.32 microg/mL. Mean plasma, CSF, synovial fluid, urine, and preocular tear film concentrations of voriconazole after long-term PO administration were 5.163 +/- 1.594 microg/mL, 2.508 +/- 1.616 microg/mL, 3.073 +/- 2.093 microg/mL, 4.422 +/- 0.8095 microg/mL, and 3.376 +/- 1.297 microg/mL, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that voriconazole distributed quickly and widely in the body; following a single IV dose, initial plasma concentrations were high with a steady and early decrease in plasma concentration. Absorption of voriconazole after PO administration was excellent, compared with absorption after IV administration. Voriconazole appears to be another option for the treatment of fungal infections in horses.     

Gentamicin concentrations in synovial fluid and joint tissues during intravenous administration or continuous intra-articular infusion of the tarsocrural joint of clinically normal horses

OBJECTIVE: To compare gentamicin concentrations achieved in synovial fluid and joint tissues during IV administration and continuous intra-articular (IA) infusion of the tarsocrural joint in horses. ANIMALS: 18 horses with clinically normal tarsocrural joints. PROCEDURE: Horses were assigned to 3 groups (6 horses/group) and administered gentamicin (6.6 mg/kg, IV, q 24 h for 4 days; group 1), a continuous IA infusion of gentamicin into the tarsocrural joint (50 mg/h for 73 hours; group 2), or both treatments (group 3). Serum, synovial fluid, and joint tissue samples were collected for measurement of gentamicin at various time points during and 73 hours after initiation of treatment. Gentamicin concentrations were compared by use of a Kruskal-Wallis ANOVA. RESULTS: At 73 hours, mean +/- SE gentamicin concentrations in synovial fluid, synovial membrane, joint capsule, subchondral bone, and collateral ligament of group 1 horses were 11.5 +/- 1.5 microg/mL, 21.1 +/- 3.0 microg/g, 17.1 +/- 1.4 microg/g, 9.8 +/- 2.0 microg/g, and 5.9 +/- 0.7 microg/g, respectively. Corresponding concentrations in group 2 horses were 458.7 +/- 130.3 microg/mL, 496.8 +/- 126.5 microg/g, 128.5 +/- 74.2 microg/g, 99.4 +/- 47.3 microg/g, and 13.5 +/- 7.6 microg/g, respectively. Gentamicin concentrations in synovial fluid, synovial membrane, and joint capsule of group 1 horses were significantly lower than concentrations in those samples for horses in groups 2 and 3. CONCLUSIONS AND CLINICAL RELEVANCE: Continuous IA infusion of gentamicin achieves higher drug concentrations in joint tissues of normal tarsocrural joints of horses, compared with concentrations after IV administration.     

Adaptations of subpalpebral lavage systems used for llamas (Lama glama) and a harbor seal (Phoca vitulina).

Subpalpebral lavage systems (SPLSs) were adapted for use in zoo llamas (Lama glama) and a wild harbor seal (Phoca vitulina) during therapy for severe ulcerative keratitis or corneal perforation. One llama presented with a melting corneal ulcer caused by Pseudomonas aeruginosa, which necessitated frequent application of a topical ophthalmic antibiotic. The lavage system was used routinely during the day and was connected to a balloon infusion system at night to allow for continuous medication administration. The ulcer healed soon after therapy was extended to include overnight treatment with the infusion system. A SPLS system was also combined with a balloon infusor during postoperative treatment of a second llama that had sustained a corneal perforation. Both llamas tolerated the infusor/lavage systems well and regained vision. One llama had minor conjunctival irritation from the SPLS that resolved quickly without treatment. Bilateral SPLS were placed in a wild harbor seal for treatment of severe ulcerative keratitis associated with Candida albicans infection. The seal tolerated the lavage systems well throughout 14 wk of their use in an aquatic environment with other seals. Partial detachment of the lavage systems from the skin of the seal occurred a few times during treatment and was easily corrected. Severe keratitis resolved with administration of antimicrobials through the lavage systems, and the seal was returned to the wild. The use of SPLSs alone or in ombination with balloon infusion systems warrants consideration for exotic, wild, and aquatic animals that cannot tolerate repetitive manual applications of topical ophthalmic medication.     

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.