Ophthalmic findings and reference values for selected ocular parameters in whooping cranes (Grus americana) and Mississippi sandhill cranes (Grus canadensis pulla).

OBJECTIVE: To establish baseline ophthalmic parameters for an endangered, semi-wild population of healthy whooping cranes (Grus americana) (WHCR) and Mississippi sandhill cranes (Grus canadensis pulla) (SACR). ANIMALS STUDIED: Eighteen WHCR and 16 SACR. PROCEDURES: Ophthalmic examination was performed by a single observer, followed by conjunctival swab collection for aerobic bacterial culture and measurement of tear production (phenol red thread test, PRTT) and corneal diameter (CD) as tolerated. Measurement of the axial globe (AG) length, anterior chamber (AC) depth, lens thickness, vitreous chamber (VC) depth, and pecten length was performed via ocular ultrasound (OUS) as tolerated. RESULTS: Eyelid cicatrization (n = 1 WHCR), keratitis (n = 2 WHCR), incipient cataracts (n = 1 WHCR, n = 4 SACR), and uveal cysts (n = 1 SACR) were identified. Twenty-one bacterial species were cultured from SACR, while 18 bacterial species were cultured from WHCR. SACR under 6 months old had increased PRTT values compared to older SACR (p = .0432). AG length and VC depth of male WHCR were greater than in female WHCR (p = .0045 and p = .0008, respectively). WHCR less than 6 months old had greater AC depth and lens thickness than WHCR over 6 months (p < .001 and p = .0013, respectively). SACR less than 6 months old had greater AC depth and lens thickness than WHCR over 6 months (p < .0001 and p < .0001, respectively). CONCLUSIONS: WHCR and SACR are amenable to complete ophthalmic examination. Age-related differences in PRTT in SACR, sexual dimorphism in WHCR, and age-related differences in AC depth and lens thickness in WHCR and SACR were identified.

Replication kinetics and cytopathic effect of feline calicivirus in feline corneal epithelial cells.

OBJECTIVE: To determine the replication kinetics and cytopathic effect (CPE) of feline calicivirus (FCV) in feline corneal epithelial cells (FCEC). ANIMALS STUDIED: Seven archived FCV isolates and one archived feline herpesvirus type 1 (FHV-1) isolate, previously obtained from eight domestic short hair cats. PROCEDURES: FCV RNA was extracted for sequencing using Illumina MiSeq, to identify three genomically diverse isolates for further testing. Following reference-based assembly, viral genomes were annotated and assessed. Superficial keratectomies were performed to isolate the corneal epithelium of cats and the cells were cultured in vitro. FCEC were infected with the three chosen FCV isolates and one FHV-1 isolate at two different multiplicity of infection ratios (MOIs, 0.1 and 0.01 PFU/cell) and virus titration was assessed at 0, 2, 6, 12, 24, and 48 h post-infection (hpi). Viral identity was confirmed by RT-qPCR. RESULTS: Three genomically diverse FCV isolates were chosen for further assessment in the FCEC model. All infections of FCEC with FCV led to visible CPE, characterized by epithelial cell rounding and detachment from the plate by 24 hpi, while FHV-1 led to visible CPE within 48 hpi. All three of the FCV isolates replicated effectively in FCEC at both 0.1 and 0.01 MOI, with a peak increase in titer approximately 12-24 hpi. CONCLUSIONS: The results support the possible role of FCV as a primary pathogen of the feline ocular surface. FCV replicates in FCEC in vitro, leading to profound CPE.

Validation of a novel rebound tonometer (Tono-Vera® Vet) in normal ex vivo rabbit eyes.

OBJECTIVE: The purpose of this study was to validate the use of the Reichert Tono-Vera® Vet tonometer rabbit setting in normal ex vivo rabbit eyes and to compare the rabbit setting to the dog, cat, and horse settings of this tonometer. PROCEDURE: Six freshly enucleated normal rabbit eyes were cannulated and connected to a fluid reservoir and physiologic monitor. Triplicate measurements were obtained with the four available settings: dog, cat, horse, and rabbit at various intraocular pressures (IOP) ranging from 5 to 80 mmHg. Bland-Altman analysis was utilized to determine bias and 95% limits of agreement for each setting. RESULTS: Linear regression equations for the dog, horse, cat, and rabbit settings were y = 0.8101x + 2.5058, y = 0.7594x - 3.4673, y = 0.6635x + 0.3021, and y = 0.8935x + 1.3295, respectively. All settings demonstrated strong positive linear trends (dog r2 = 0.9644, horse r2 = 0.9456, cat r2 = 0.9309, and rabbit r2 = 0.9558). Bland-Altman plots revealed that the average bias and 95% limits of agreement (mmHg) were -4.73, -12.65, -12.86 and -2.73 and (-15.31, 5.86), (-29.03, 3.74), (-25.67, -0.05), and (-12.21, 6.76) for the dog, horse, cat, and rabbit settings, respectively. CONCLUSION: The Tono-Vera® Vet rabbit setting provided the most accurate and precise measurements compared with the other settings, but slightly underestimated actual IOP, especially as IOP was increased. This tonometer, using the rabbit setting, is likely to be appropriate for the estimation of IOP in rabbits with the appropriate correction formula applied.

Calibration of a novel rebound tonometer (Tono-Vera® vet) in the porcine eye.

OBJECTIVE: Porcine models of ocular disease are becoming increasingly utilized. A recently commercialized ocular tonometer, the Reichert Tono-Vera® Vet, has not been evaluated for use in pigs. The purpose of this study was to calibrate this device for use in porcine eyes and to determine which settings are most appropriate for use in pigs. PROCEDURE: The anterior chambers of five freshly enucleated normal porcine eyes were cannulated then connected to a reservoir of balanced salt solution and a physiologic monitor. Triplicate measurements were obtained with the four available settings: dog, cat, horse, and rabbit at intraocular pressures ranging from 5- to 80 mmHg. Bland-Altman analysis was utilized to determine bias and 95% limits of agreement for each setting. RESULTS: There was a strong positive linear regression trend for all settings (dog r2  = 0.986, horse r2  = 0.947, cat r2  = 0.977, and rabbit r2  = 0.982). The linear regression equations for the dog, horse, cat, and rabbit setting were y = 1.0168x - 2.6128, y = 0.8743x - 3.4959, y = 0.9394x - 7.3188, and y = 1.1082x - 3.4077. The average bias and 95% limits of agreement for dog, horse, cat, and rabbit settings were - 2.00, -8.32, -9.58, and 0.57 mmHg, and (-7.52, 3.53), (-19.00, 2.37), (-16.66, -2.50), and (-7.79, 8.93), in mmHg. CONCLUSION: The Tono-Vera® Vet dog setting was most accurate and precise setting compared to true intraocular pressures. This setting is likely to be appropriate for in vivo use in pigs, with the appropriate correction formula applied.

Relationship between the bacterial ocular surface microbiota and outcomes for cats with feline herpesvirus type 1 ocular surface disease.

OBJECTIVE: Feline herpesvirus 1 (FHV-1) causes ocular surface disease in domestic cats. The purpose of this study was to assess the relationship between bacterial ocular surface microbiota and outcomes for cats with FHV-1 ocular surface disease. ANIMALS STUDIED: Twenty-two shelter-housed cats with confirmed FHV-1 ocular surface disease. PROCEDURES: Animals were grouped according to FHV-1 shedding and ocular clinical scores following intervention: worsened outcome (WorOut, n = 11) or improved outcome (ImpOut, n = 11). Scoring and conjunctival sampling were completed on Days 1 and 8 of twice daily antiviral treatment. Bacterial DNA was extracted and submitted for 16S rRNA gene sequencing. Real-time polymerase chain reaction was performed for selected bacterial species. Overall DNA concentration between groups was assessed. RESULTS: Bacterial microbiota relative abundance composition was significantly different between ImpOut and WorOut groups (weighted UniFrac p = .006). Alpha diversity was significantly higher in the ImpOut group compared with the WorOut group (Shannon p = .042, Simpson's p = .022, Pielou's p = .037). Differences in the relative abundance of various phyla and species were detected between groups. Total DNA concentration was higher in the WorOut group compared with the ImpOut group (p = .04). Feline GAPDH (p = .001) and Bilophila wadsworthia (p = .024) copy number was significantly higher in the ImpOut group compared with the WorOut group. CONCLUSIONS: The results highlight the important relationship between the bacterial ocular surface microbiota and FHV-1 infection outcomes in cats treated with antiviral medications. Low bacterial species diversity, higher overall DNA (presumed predominantly bacterial) load, and certain bacterial phyla/species were associated with poor outcomes for cats with FHV-1 ocular disease.

Effect of multiple head positions on intraocular pressure in healthy, anesthetized horses during hoisting.

OBJECTIVE: To evaluate changes in intraocular pressure (IOP) with variable head position in healthy, anesthetized horses in hoisted inversion and to assess the influence of various cofactors (age, sex, body weight, body condition score, and neck length) on IOP changes during hoisting. ANIMALS STUDIED: Seventeen healthy adult horses without significant ocular abnormalities. PROCEDURES: Subjects were administered intravenous xylazine/butorphanol premedication and ketamine/midazolam induction with xylazine/ketamine boluses for anesthetic maintenance. While hoisted, IOP was measured in triplicate for each eye via rebound tonometry (TonoVet) at neutral head position (ie, eyes level with the withers), at multiple 5 cm increments above and below neutral (-20 cm through +20 cm) using foam pads for head support, and with eyes above heart level via manual support. RESULTS: In hoisted positions, IOP ranged from 18 to 51 mmHg. Intraocular pressure significantly decreased with head position elevated ≥+15 cm from neutral and significantly increased when lowered ≤-5 cm from neutral. Neck length significantly influenced IOP (P = .0328) with linear regression indicating a median (range) increase of 0.244 (0.034-0.425) mmHg in IOP for every 1 cm increase in neck length. Age, sex, breed, body weight, body condition score, and eye (OD vs OS) did not significantly influence IOP. Intraocular pressure only varied significantly between eyes at +10 cm above neutral (OS > OD, 1.7 ± 0.6 mm Hg, P = .0044). CONCLUSIONS: Intraocular pressure in healthy, anesthetized horses varies with head position during hoisting; increased neck length may be associated with larger changes in IOP during hoisting.

Ocular findings in a group of healthy captive leopard geckos.

OBJECTIVE: Leopard Geckos (Eublepharis macularius) are popular pets and can be affected by a range of ocular disorders. Our objective was to report ocular findings in a group of healthy captive leopard geckos and to establish reference ranges for commonly performed ocular diagnostic tests. ANIMALS STUDIED: Twenty-six healthy male geckos aged 1 year old (n = 4) and >2 years old (n = 22). PROCEDURES: All animals underwent ophthalmic examination, corneal esthesiometry, modified Schirmer tear test (mSTT), rebound tonometry, conjunctival bacterial aerobic and fungal culture, and measurement of ocular dimensions. Student's t test was used to compare values of corneal esthesiometry, tonometry and mSTT between groups. Multiple correlations were assessed by Pearson correlation coefficient. RESULTS: All animals had a normal ocular examination. Tear production as measured with a mSTT (mean ± SD) technique was 3.1 ± 1.3 mm/min and tonometry values (mean ± SD) were 8.2 ± 1.7 mm Hg. Corneal touch threshold (median, range) was 4.4 cm, 2.5-5.0. Younger animals had a significantly increased corneal sensitivity compared to older animals (P = .0383). Results of culture showed no growth for fungal organism in any animals. Conjunctival bacterial isolation rates were low, with only 7/26 samples positive for nine bacterial species. CONCLUSIONS: Leopard geckos are amenable to ophthalmic examination and ocular diagnostic database testing with minimal manual restraint.

Advances in Diagnostics and Treatments in Horses and Foals with Gastric and Duodenal Ulcers.

Equine gastric ulcer syndrome (EGUS) primarily describes ulceration in the terminal esophagus, nonglandular squamous mucosa, glandular mucosa of the stomach, and proximal duodenum. EGUS is common in all breeds and ages of horses and foals. This article focuses on the current terminology for EGUS, etiologies and pathogenesis for lesions in the nonglandular and glandular stomach, diagnosis, and a comprehensive approach to the treatment and prevention of EGUS in adult horses and foals.

Surveillance for feline herpesvirus type 1 mutation and development of resistance in cats treated with antiviral medications.

Feline herpesvirus type 1 (FHV-1) commonly causes ocular surface disease in cats and is treated with antiviral medications targeting viral DNA polymerase (UL30/42). Herein, we describe a method to assess the FHV-1 genome for mutation development and to assess the functional impact of mutations, if present. Fourteen shelter-housed domestic cats with FHV-1 ocular surface disease were assigned to one of four treatment groups: placebo (n = 3), cidofovir 0.5% ophthalmic solution (n = 3), famciclovir oral solution (n = 5), or ganciclovir 0.15% ophthalmic solution (n = 3). Swabs were collected before (day 1) and after (day 8) 1 week of twice-daily treatments to isolate viable FHV-1. Viral DNA was extracted for sequencing using Illumina MiSeq with subsequent genomic variant detection between paired day 1 and day 8 isolates. Plaque reduction assay was performed on paired isolates demonstrating non-synonymous variants. A total of 171 synonymous and 3 non-synonymous variants were identified in day 8 isolates. No variants were detected in viral UL23, UL30, or UL42 genes. Variant totals were not statistically different in animals receiving antiviral or placebo (p = 0.4997). A day 8 isolate from each antiviral treatment group contained a single non-synonymous variant in ICP4 (transcriptional regulator). These 3 isolates demonstrated no evidence of functional antiviral resistance when IC50 was assessed. Most (10/14 pairs) day 1 and 8 viral isolate pairs from the same host animal were near-identical. While functional variants were not detected in this small sample, these techniques can be replicated to assess FHV-1 isolates suspected of having developed resistance to antiviral medications.

In vitro efficacy of ganciclovir against feline herpesvirus type 1 and assessment of ocular tolerability in healthy cats.

OBJECTIVES: Feline herpesvirus-1 (FHV-1) is a prevalent cause of ocular disease in cats and limited topical options for treatment currently exist. The first objective of this study was to confirm the efficacy of ganciclovir against FHV-1 in vitro. The second objective was to assess the safety and ocular tolerability of topically applied ganciclovir eye gel (GEG) in healthy cats. METHODS: FHV-1 was used to infect tissue culture wells covered in maximally confluent Crandall-Rees feline kidney cells prior to the addition of three molarities of ganciclovir (8.9 µM, 17.8 µM and 89 µM) before being incubated for 48 h. Ganciclovir efficacy in vitro was then assessed using standard plaque reduction assay. Commercially available GEG (0.15%) was applied q8h to one randomly chosen eye of four healthy cats for 7 days. Commercially available lubricating eye gel (LEG) was applied to the opposite eye q8h. Complete blood counts (CBCs), blood chemistry panels (CHEM) and urinalysis (UA) were performed on all cats before and after the study period. Ocular lesions were assessed daily using a standardized scheme. RESULTS: Ganciclovir led to a significant reduction in FHV-1 plaque number, area and diameter at all tested molarities in vitro. The highest molarity assessed (89 µM) caused a 100% reduction in viral plaque number. There was no significant difference in ocular lesion scores between eyes receiving GEG and LEG. Animals remained healthy throughout the study period with CBC, CHEM and UA showing no clinically significant alterations. CONCLUSIONS AND RELEVANCE: Based on the in vitro results, ganciclovir appears to be effective against FHV-1 in vitro. When applied q8h as a commercial 0.15% gel to a small group of cats with normal eyes, this medication was well tolerated. Taken together, these data suggest this medication warrants further investigation in cats with ocular disease caused by FHV-1.

Inter-user and intra-user variation of two tonometers in horses.

BACKGROUND: It is currently unknown which of the two devices most commonly used in equine ophthalmology for intraocular pressure (IOP) estimation demonstrates the lowest inter-user and intra-user variation. OBJECTIVES: To assess the inter-user and intra-user variation of two tonometers in sedated and unsedated horses. STUDY DESIGN: Randomised masked cross-over trial. METHODS: Four examiners used the rebound (ICare® TonoVet) and applanation (TonoPen® ) tonometers to measure the intraocular pressure (IOP) in triplicate in 10 normal horses before and after sedation with xylazine. For inter-user variation, coefficient of variation (CV) values were calculated from the mean of each examiner for each condition combination. For intra-user variation, CV values were calculated from the individual measurements of each examiner for each condition combination. CV values were also assessed in relation to other variables using ANOVA. RESULTS: The rebound tonometer was found to have lower inter-user (15.4% vs 21.7%, P = .01) and intra-user (9.1% vs 16.1%, P < .0001) variation in unsedated horses and lower intra-user (8.4% vs 14.7%, P < .0001) variation in sedated horses than the applanation tonometer. Both instruments had similar inter-user variation in sedated horses. For the rebound tonometer, sedation did not affect inter-user or intra-user variation, but for the applanation tonometer inter-user variation was lowest while horses were sedated (16.0% vs 21.7%, P = .03). No other variable assessed was found to have an effect on IOP. MAIN LIMITATIONS: No animals with ocular disease were included in this study. CONCLUSIONS: The rebound tonometer may be the preferred instrument to minimise intra-user and inter-user variation for IOP measurement in unsedated horses. The rebound tonometer is also likely to be the preferred instrument to minimise intra-user variation in sedated horses. If the applanation tonometer is used to perform IOP measurement in horses, it is recommended that this is performed while horses are sedated to minimise inter-user variation for this instrument.