Detection of Mycoplasma spp. and feline calicivirus in cats with ocular surface disease.

OBJECTIVE: To survey the prevalence of pathogens in shelter-housed cats with active ocular surface disease (OSD). ANIMALS STUDIED: A total of 255 shelter-housed domestic cats with evidence of active OSD. No normal, unaffected cats were sampled. PROCEDURE(S): OSD scoring was performed on cats with active OSD. Combined oropharyngeal/conjunctival swabs were submitted for rt-PCR/PCR for feline herpesvirus (FHV-1), feline calicivirus (FCV), Chlamydia spp. (CHL), Bordetella bronchiseptica (BORD), and Mycoplasma spp. (MYC). RESULTS: Pathogens were detected as follows: 76.4% (195/255) MYC, 57.6% (147/255) FHV-1, 42.7% (109/255) FCV, 26.7% (68/255) CHL, and 5.5% (14/255) BORD. Monoinfections affected 21.1% (54/255) animals, with MYC being the most common monoinfection (12.5%, 32/255), followed by FHV-1 (4.7%, 12/255), followed by CHL (2.4%, 6/255), followed by FCV (1.6%, 4/255), with no animals having a BORD monoinfection. Dual infections affected 36.4% of animals (93/255), with MYC detected in 30.1% (77/255) dual infections and FCV detected in 12.9% (33/255) dual infections. Dual infections with MYC and FCV together were detected in 9.8% (25/255) animals. Many animals (35.3%, 90/255) were found to be affected by 3 or more pathogens, and 7.1% (18/255) animals had no pathogens detected. OSD scores were not influenced by any variable assessed, including the number and type of pathogens detected. CONCLUSION: MYC, FHV-1, FCV, and CHL were commonly detected in this group of animals with OSD. Both MYC and FCV (alone or in combination with each other) were detected in multiple animals with active OSD, supporting prior evidence that either may independently act as a primary ocular surface pathogen.

Ocular examination findings and selected ophthalmic diagnostic tests in African green monkeys (Chlorocebus aethiops sabaeus).

OBJECTIVE: To document ocular lesions and establish ophthalmic diagnostic test reference values in a colony of African green monkeys (Chlorocebus aethiops sabaeus). ANIMALS STUDIED: Fifty one geriatric (GAGM, 19-30 years old), 10 adult (AAGM, 5-9 years old) and 10 juvenile (JAGM, <2 years old) African green monkeys housed in a single Caribbean research colony. PROCEDURES: Ocular biomicroscopy, indirect fundoscopy, Schirmer tear test (STT), rebound tonometry (TonoVet®) and corneal fluorescein staining were performed. Mixed ANCOVA tests were performed to compare STT and IOP between groups. RESULTS: Common ocular lesions in GAGM included vitreal degeneration (27/51, 51/102 eyes) and cataracts (21/51, 32/102 eyes). Vitreal degeneration was also common in AAGM (8/10, 16/20 eyes) and infrequent in JAGM (3/10, 6/20 eyes). Cataracts were not present in any JAGM or AAGM. All eyes in all three groups had perilimbal corneal pigmentation and faint lace-like anterior corneal stromal opacification. Median (range) STT values were 16.0 (18) mm/min in GAGM. Mean (SD) STT values were 14.2 (4.6) mm/min in AAGM, and 8.9 (3.4) mm/min in JAGM. Median (range) IOP values were 16.5 (27) mmHg in GAGM. Mean (SD) IOP values were 18.0 (2.8) mmHg in AAGM, and 14.1 (2.2) mmHg in JAGM. JAGM had significantly lower STT and IOP values compared to AAGM (p = .0449, .0057, respectively) and GAGM (p = .0002, .0130, respectively). CONCLUSIONS: Spontaneous ocular lesions were common in geriatric monkeys in this research colony. IOP and STT values were lower in juvenile African green monkeys relative to adult or geriatric animals.

Longitudinal intraocular pressure measurements in Whooping cranes (Grus americana) and Mississippi-Sandhill cranes (Grus canadensis pulla).

OBJECTIVE: To assess intraocular pressure (IOP) development in cranes and determine the impact of age, weight, species, head position, and sex. ANIMALS STUDIED: Whooping cranes (WC) (Grus americana), and Mississippi-sandhill cranes (MSC) (Grus canadensis pulla). PROCEDURES: Chicks were manually restrained on days 1-3, 7, 21, 35, 60, 75, and 120 for routine examinations. IOP was opportunistically measured utilizing the Tonovet Plus® in D setting with the head above the heart (AH) and below the heart (BH). Values were also obtained longitudinally in adults (>120 days old) upon presentation in 1 year. RESULTS: Intraocular pressure was highly correlated with age and weight in chicks. For every kilogram gained, IOP increased 2.46 ± 0.08 mmHg in WC and 2.66 ± 0.11 mmHg in MSC. Once hatched, IOP increased 1.13 ± 0.04 mmHg in WC and 0.87 ± 0.04 mmHg in MSC every 10 days. IOP was similar to adults at 120 days of age. In adult WC, mean IOP AH was 24.0 ± 0.4 mmHg, and BH was 27.9 ± 0.4 mmHg, there was a significant difference regarding head positioning and sex, females (25.3 ± 0.4 mm Hg) had lower IOP than males (26.5 ± 0.4 mmHg). In adult MSC, mean IOP AH was 20.7 ± 0.4 mmHg, and BH was 24.6 ± 0.4 mmHg. The difference between head positioning was significant. CONCLUSIONS: This study documents the correlation between IOP and weight or age during early development in cranes, as well as the importance of head positioning.

Risk factors for corneal conjunctival grafting failure in dogs at four referral centers in the southern United States (203 patients): 2015-2021.

OBJECTIVE: To report risk factors associated with conjunctival graft failure in dogs at four referral specialty centers. PROCEDURES: Records of 203 dogs (229 eyes) undergoing conjunctival graft repair of ulcerative keratitis at four hospitals from 2015 to 2021 were reviewed. Success was defined as full graft integration with globe retention at the last postoperative evaluation; vision status was reported separately. Factors assessed included patient signalment, ophthalmic examination findings, surgical factors, and follow-up information. RESULTS: Conjunctival graft failure occurred in 11% (25/229) of eyes and was significantly associated with ulcer depth, with corneal perforations having increased odds of graft failure compared with descemetoceles (odds ratio [OR] = 3.22, 95% confidence interval [CI] 1.11-9.32; p = .03) and stromal ulcers (OR = 10.89 [95% CI 1.38-86.18], p = .02). Brachycephalic dogs were significantly more likely than non-brachycephalic dogs to experience graft failure (OR = 5.02 [95% CI 1.42-17.74], p < .01). Surgery on the opposite eye relative to surgeon handedness was significantly associated with an increased risk of graft failure (OR 4.28 [95% CI 1.53-11.94], p < .01). The use of 7-0 and 8-0 suture versus 9-0 (p = .03) and the use of a combined simple continuous and interrupted pattern (p = .03) were significantly associated with an increased risk of graft failure. At the last follow-up, (median 61.5 days), 87% of dogs were visual. CONCLUSIONS: Corneal perforations, surgery on the opposite eye relative to surgeon handedness, suture size and pattern, and brachycephalic conformation were significantly associated with an increased risk of graft failure in this study. These characteristics can be considered when determining prognosis for ulcerative keratitis requiring surgical intervention.

Phylogenomic assessment of 23 equid alphaherpesvirus 1 isolates obtained from USA-based equids.

BACKGROUND: Equid alphaherpesvirus 1 (EHV-1) is a global viral pathogen of domestic equids which causes reproductive, respiratory and neurological disease. Few isolates acquired from naturally infected USA-based hosts have been fully sequenced and analyzed to date. An ORF 30 (DNA polymerase) variant (A2254G) has previously been associated with neurological disease in host animals. The purpose of this study was to perform phylogenomic analysis of EHV-1 isolates acquired from USA-based hosts and compare these isolates to previously sequenced global isolates. METHODS: EHV-1 was isolated from 23 naturally infected USA-based equids (6 different states, 15 disease outbreaks) with reproductive (22/23) or neurological disease (1/23). Following virus isolation, EHV-1 DNA was extracted for sequencing using Illumina MiSeq. Following reference-based assembly, whole viral genomes were annotated and assessed. Previously sequenced EHV-1 isolates (n = 114) obtained from global host equids were included in phylogenomic analyses. RESULTS: The overall average genomic distance was 0.0828% (SE 0.004%) for the 23 newly sequenced USA isolates and 0.0705% (SE 0.003%) when all 137 isolates were included. Clade structure was predominantly based on geographic origin. Numerous nucleotide substitutions (mean [range], 179 [114-297] synonymous and 81 [38-120] non-synonymous substitutions per isolate) were identified throughout the genome of the newly sequenced USA isolates. The previously described ORF 30 A2254G substitution (associated with neurological disease) was found in only one isolate obtained from a host with non-neurological clinical signs (reproductive disease), six additional, unique, non-synonymous ORF 30 substitutions were detected in 22/23 USA isolates. Evidence of recombination was present in most (22/23) of the newly sequenced USA isolates. CONCLUSIONS: Overall, the genomes of the 23 newly sequenced EHV-1 isolates obtained from USA-based hosts were broadly similar to global isolates. The previously described ORF 30 A2254G neurological substitution was infrequently detected in the newly sequenced USA isolates, most of which were obtained from host animals with reproductive disease. Recombination was likely to be partially responsible for genomic diversity in the newly sequenced USA isolates.

A review of evidence-based management of infectious ocular surface disease in shelter-housed domestic cats.

Infectious ocular surface disease (IOSD) is a common problem in shelter-housed domestic cats and has a widespread negative impact on animal welfare. While the common etiological agents are well-described, addressing IOSD in large groups of animals presents a management challenge to the clinician and logistical challenges to shelter employees. Treatments, diagnostics, and prevention strategies that are effective in privately owned or experimental animals may be impractical or ineffective in the shelter environment. This review article focuses on the relative prevalence of etiological agents in feline IOSD, practical diagnostic testing protocols, prevention strategies, and treatment of IOSD in shelter-housed cats. Discrepancies between experimental laboratory-based studies and clinical trials assessing therapeutics for treatment of feline herpes virus are highlighted. Further high-quality clinical trials are necessary to determine optimal preventative and therapeutic protocols for IOSD in shelter-housed cats.

Efficacy of perioperative 2% dorzolamide ophthalmic solution for reduction of postoperative ocular hypertension in dogs following routine phacoemulsification surgery: A prospective clinical trial.

OBJECTIVE: To evaluate the efficacy of 2% dorzolamide ophthalmic solution for reduction of postoperative ocular hypertension (POH) following routine phacoemulsification surgery in dogs. ANIMALS STUDIED: Thirty one dogs (53 eyes) with naturally occurring cataracts undergoing routine phacoemulsification surgery. PROCEDURE(S): A prospective, double-masked, randomized, placebo-controlled study design was utilized. Dogs received 2% dorzolamide ophthalmic solution or saline 1 h prior to surgery then three times daily for 21 days postoperatively in the operated eye(s). Intraocular pressure (IOP) was recorded 1 h prior to surgery and 3 h, 7 h, 22 h, 1 week and 3 weeks postoperatively. Statistical analyses were performed using chi-squared and Mann-Whitney U test with a significance level of p < .05. RESULTS: Postoperative ocular hypertension (IOP ≥25 mmHg, <24 h after surgery) occurred in 28/53 (52.8%) eyes. There was significant reduction in the incidence of POH for eyes receiving dorzolamide (10/26 (38.4%) eyes) versus eyes receiving placebo (18/27 (66.7%) eyes) (p = .0384). Animals were followed for a median of 163 days after surgery. Thirty-seven (37/53 (69.8%)) eyes were visual at final examination and 3/53 (5.7%) globes were enucleated postoperatively. At last follow-up, there was no difference in visual status (p = .9280), need for topical IOP lowering medication (p = .8319) or incidence of glaucoma (p = .5880) based on treatment group. CONCLUSIONS: Perioperative administration of topical 2% dorzolamide reduced the incidence of POH after phacoemulsification in the dogs studied. However, this was not associated with differences in visual outcome, incidence of glaucoma or need for IOP-lowering medications.

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.

Evaluation of compounded cidofovir, famciclovir, and ganciclovir for the treatment of feline herpesvirus ocular surface disease in shelter-housed cats.

OBJECTIVE: To assess the efficacy of compounded cidofovir, famciclovir, and ganciclovir for the treatment of feline herpesvirus type 1 (FHV-1) ocular surface disease. ANIMALS STUDIED: 132 shelter-housed cats qPCR positive for FHV-1. PROCEDURES: A masked placebo-controlled study design was utilized. Animals were enrolled in one of four treatment groups: topical ocular placebo + oral placebo (n = 32), compounded cidofovir 0.5% ophthalmic solution + oral placebo (n = 32), compounded famciclovir oral solution (90 mg/kg) + topical ocular placebo (n = 32), and compounded ganciclovir 0.15% ophthalmic solution + oral placebo (n = 36). Cats were treated with each medication twice daily for 7 days and were evaluated on Day 1 and Day 8 using an ocular scoring system, body weight, and qPCR for FHV-1 viral load. RESULTS: Cidofovir significantly decreased viral load from Day 1 to Day 8 compared with placebo (p = .024). Neither famciclovir nor ganciclovir decreased viral load compared with placebo (p = .14, p = .41). There was no significant improvement of ocular scores for any drug group compared with placebo (p = .62). In all groups, 65%-75% of cats improved from Day 1 to Day 8. Juvenile cats had a significant increase in weight gain compared with placebo for cidofovir (p = .025) and ganciclovir (p = .023). All corneal ulcers in placebo animals failed to heal whereas 77% of ulcers in antiviral group animals healed. CONCLUSIONS: Topical ophthalmic cidofovir significantly decreased ocular FHV-1 viral shedding and increased weight gain in juvenile cats. Ganciclovir increased weight gain in juvenile cats. Compounded famciclovir demonstrated limited efficacy for the treatment of FHV-1 ocular surface disease in shelter-housed cats.

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.

Successful Surgical Removal of the Crystalline Lenses in a Black-Necked Swan (Cygnus melancoryphus).

A captive-bred, adult, male, black-necked swan (Cygnus melancoryphus) was presented for evaluation of apparent vision loss due to cataract formation of an unknown duration. The animal was having difficulty navigating its enclosure, and lenticular opacities had been previously noted in both eyes. On examination, bilateral hypermature cataracts were diagnosed. Following preoperative diagnostic testing, surgical removal of the crystalline lenses in both eyes was performed using minor modifications of standard techniques. Follow-up examination and behavioral observation at 60 days postsurgery indicated that vision had been successfully restored without complications. We conclude that successful surgical removal of cataracts is possible in this species using modifications of standard techniques.

The effect of topical ophthalmic proparacaine, fluorescein, and tropicamide on subsequent bacterial cultures in healthy dogs.

OBJECTIVE: To determine whether tropicamide, fluorescein, and proparacaine applied topically before sample collection affect the quantity or species of bacteria isolated via aerobic culture. ANIMALS STUDIED: 12 female adult research beagle cross-breed dogs. PROCEDURES: A conjunctival swab was taken before and after the sequential application of proparacaine, tropicamide, and fluorescein to the same eye (P/T/F) with a five-minute gap between medications. Paired swabs were submitted for aerobic culture. Bacterial enumeration was performed using the spread plate method. Following a one-week washout period, the procedure was repeated using balanced salt solution (BSS). Following a second one-week washout period, the experiment was repeated using ofloxacin 0.3% solution. Colony counts were compared using one-way ANOVA and Tukey post hoc comparison. Bacterial species reduction was compared using a Friedman rank test and Dunn's method. RESULTS: The bacterial colony count for P/T/F and BSS was significantly higher than the ofloxacin group (p = 0.0052, p = 0.0022). There was no significant difference for colony counts between P/T/F and BSS (p = 0.9295). The most frequently isolated bacteria included: Psychrobacter spp., Staphylococcus spp., Corynebacterium spp., and Streptococcus spp. The bacterial species reduction for P/T/F and BSS was significantly lower than for ofloxacin (p < 0.0001, p = 0.0160). There was no significant difference for species reduction between P/T/F and BSS (p = 0.3749). CONCLUSIONS: The application of proparacaine, tropicamide, and fluorescein did not significantly decrease the amount or species of bacteria isolated from the conjunctiva in this canine population. The application of these solutions prior to ocular swab collection in healthy dogs is unlikely to affect subsequent culture results.

Current ocular microbiome investigations limit reproducibility and reliability: Critical review and opportunities.

Enthusiasm for research describing microbial communities using next-generation sequencing (NGS) has outpaced efforts to standardize methodology. Without consistency in the way research is carried out in this field, the comparison of data between studies is near impossible and the utility of results remains limited. This holds true for bacterial microbiome research of the ocular surface, and other sites, in both humans and animals. In addition, the ocular surface remains under-explored when compared to other mucosal sites. Low bacterial biomass samples from the ocular surface lead to further technical challenges. Taken together, two major problems were identified: (1) Normalization of the workflow in studies utilizing NGS to investigate the ocular surface bacteriome is necessary in order to propel the field forward and improve research impact through cross-study comparisons. (2) Current microbiome profiling technology was developed for high bacterial biomass samples (such as feces or soil), posing a challenge for analyses of samples with low bacterial load such as the ocular surface. This article reviews the challenges and limitations currently facing ocular microbiome research and provides recommendations for minimum reporting standards for veterinary ophthalmologists and clinician scientists to limit inter-study variation, improve reproducibility, and ultimately render results from these studies more impactful. The move toward normalization of methodology will expedite and maximize the potential for microbiome research to translate into meaningful discovery and tangible clinical applications.

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.

Development of and Validity Evidence for a Canine Ocular Model for Training Novice Veterinary Students to Perform a Fundic Examination.

Indirect fundoscopy is challenging for novice learners, as patients are often intolerant of the procedure, impeding development of proficiency. To address this, we developed a canine ocular simulator that we hypothesized would improve student learning compared to live dogs. Six board-certified veterinary ophthalmologists and 19 second-year veterinary students (novices) performed an indirect fundic examination on the model and live dog. Prior to assessment, novices were introduced to the skill with a standardized teaching protocol and practiced (without feedback) with either the model (n = 10) or live dog (n = 9) for 30 minutes. All participants evaluated realism and usefulness of the model using a Likert-type scale. Performance on the live dog and model was evaluated in all participants using time to completion of task, performance of fundic examination using a checklist and global score, identification of objects in the fundus of the model, and evaluation of time spent looking at the fundus of the model using eye tracking. Novices (trained on simulator or live dogs) were compared in fundic examination performance on the live dog and identification of shapes in the model. In general, experts performed the fundic examination faster (p ≤ .0003) and more proficiently than the novices, although there were no differences in eye tracking behavior between groups (p ≥ .06). No differences were detected between training on simulator versus live dog in development of fundoscopy skills in novices (p ≥ .20). These findings suggest that this canine model may be an effective tool to train students to perform fundoscopy.

Ophthalmic Evaluation of Raptors Suffering From Ocular Trauma.

Ocular problems are often associated with traumatic injury in raptors. A comprehensive evaluation, including a complete ophthalmic examination, is vital in determining the patient's overall health and suitability for release. Steps for conducting ocular examination and diagnostic testing in raptors is discussed. Additionally, common clinical findings after trauma, as well as the mechanisms by which ocular injury occurs, are outlined. An overview of medical treatments recommended for commonly diagnosed ocular diseases and the utility of ancillary diagnostic procedures is also presented.

Genomic analysis for virulence determinants in feline herpesvirus type-1 isolates.

Feline herpesvirus type 1 (FHV-1) is a widespread cause of respiratory and ocular disease in domestic cats. A spectrum of disease severity is observed in host animals, but there has been limited prior investigation into viral genome factors which could be responsible. Stocks of FHV-1 were established from oropharyngeal swabs obtained from twenty-five cats with signs of infection housed in eight animal shelters around the USA. A standardized numerical host clinical disease severity scoring scheme was used for each cat from which an isolate was obtained. Illumina MiSeq was used to sequence the genome of each isolate. Genomic homogeneity among isolates was relatively high. A general linear model for fixed effects determined that only two synonymous single nucleotide polymorphisms across two genes (UL37/39) in the same isolate (from one host animal with a low disease severity score) were significantly associated (p ≤ 0.05) with assigned host respiratory and total disease severity score. No variants in any isolate were found to be significantly associated with assigned host ocular disease severity score. A concurrent analysis of missense mutations among the viral isolates identified three genes as being primarily involved in the observed genomic variation, but none were significantly associated with host disease severity scores. An ancestral state likelihood reconstruction was performed and determined that there was no evidence of a connection between host disease severity score and viral evolutionary state. We conclude from our results that the spectrum of host disease severity observed with FHV-1 is unlikely to be primarily related to viral genomic variations, and is instead due to host response and/or other factors.

Environmental relative humidity affects Schirmer tear test results in normal dogs.

OBJECTIVE: To investigate the effects of environmental temperature and relative humidity on tear test results in dogs. ANIMALS STUDIED: 22 normal dogs. PROCEDURES: Routine Schirmer tear tests (STT1) and phenol red thread tests (PRTT) were administered to normal dogs in four environmental conditions with different relative humidity (RH) and temperature (T). Environmental conditions (mean ± SD) included the following: normal indoor (RH = 49 ± 4%, T = 72 ± 2°F), outdoor (RH = 62 ± 11%, T = 85 ± 6°F), indoor high humidity (RH = 85 ± 6%, T = 71 ± 1°F), and indoor low humidity (RH = 39 ± 2%, T = 76 ± 1°F). ANCOVA models were used to assess for significant associations between tear test readings and RH/T (P < .05). RESULTS: A significant, direct positive relationship between RH and STT1 results was detected (y = 0.04909*X + 19.11, P = .006) but not between T and STT1. There was no significant relationship between PRTT and RH or T. CONCLUSIONS: These results suggest that STT1 but not PRTT is affected by RH and that T does not influence STT1 or PRTT results in normal dogs. RH is unlikely, however, to cause clinically significant effects on STT1 readings in normal dogs when performed in average indoor conditions. Future studies should investigate the influence of RH on STT1 readings obtained in dogs with keratoconjunctivitis sicca.

Schirmer strip wetting length varies by commercial manufacturer.

OBJECTIVE: To determine whether wetting length of Schirmer tear test (STT) strips varies by commercial manufacturer of test. ANIMALS STUDIED: Ten normal female beagle cross-breed dogs. PROCEDURES: Schirmer tear test strips from four commercial manufacturers were used to assess wetting lengths in-vitro when exposed to a set volume of water over 1 minute. Digital photography was used to assess the total surface area of Schirmer strips from each manufacturer. Schirmer tear test type 1 was performed on normal dogs over 1 minute with STT strips from the same four commercial manufacturers in a randomized crossover design. ANOVA was used to detect differences between strips from different manufacturers. RESULTS: Significant differences in wetting length were found between STT strips from different manufacturers in-vitro (P < .0001) and in-vivo (P < .0001). STT wetting lengths for the in-vitro experiments (mean ± SD) were 21.2 ± 0.8 mm (Amcon Laboratories), 27.8 ± 2.7 mm (Merck Animal Health), 30.0 ± 1.7 mm (HUB pharmaceuticals), and 31.5 ± 2.3 mm (Gulden Ophthalmics). STT wetting lengths for the experiments in live dogs (mean ± SD) were 17.5 ± 3.4 mm (Amcon Laboratories), 24.8 ± 3.9 mm (Merck Animal Health), 26.5 ± 3.7 mm (HUB pharmaceuticals), and 24.0 ± 3.9 mm (Gulden Ophthalmics). The surface area of STT strips were found to vary between different manufacturers. CONCLUSIONS: Schirmer tear test type 1 results in dogs are affected by choice of STT strip commercial manufacturer. It is proposed that guidelines are created for the standardization of future STT strip production.

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.