Assessment of Cidofovir for Treatment of Ocular Bovine Herpesvirus-1 Infection in Cattle Using an Ex-Vivo Model.

Bovine herpesvirus-1 (BoHV-1) infection contributes to keratoconjunctivitis, respiratory disease, and reproductive losses in cattle. The objective of this study was to determine the most appropriate ophthalmic antiviral agent for BoHV-1 inhibition using in-vitro culture and novel ex-vivo bovine corneal modeling. Half-maximal inhibitory concentrations of BoHV-1 were determined for cidofovir, ganciclovir, idoxuridine, and trifluridine via in-vitro plaque reduction assays. In-vitro cytotoxicity was compared amongst these compounds via luciferase assays. Trifluridine and cidofovir were the most potent BoHV-1 inhibitors in vitro, while trifluridine and idoxuridine were the most cytotoxic agents. Therefore, cidofovir was the most potent non-cytotoxic agent and was employed in the ex-vivo corneal assay. Corneoscleral rings (n = 36) from fresh cadaver bovine globes were harvested and equally divided into an uninfected, untreated control group; a BoHV-1-infected, untreated group; and a BoHV-1-infected, cidofovir-treated group. Virus isolation for BoHV-1 titers was performed from corneal tissue and liquid media. Histologic measurements of corneal thickness, epithelial cell density, and tissue organization were compared between groups. Substantial BoHV-1 replication was observed in infected, untreated corneas, but BoHV-1 titer was significantly reduced in cidofovir-treated (1.69 ± 0.08 × 103 PFU/mL) versus untreated (8.25 ± 0.25 × 105 PFU/mL, p < 0.0001) tissues by day 2 of culture. No significant differences in histologic criteria were observed between groups. In conclusion, cidofovir warrants further investigation as treatment for BoHV-1 keratoconjunctivitis, with future studies needed to assess in-vivo tolerability and efficacy.

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.

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.

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.

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.

Systemic humoral immunity in beef bulls following therapeutic vaccination against Tritrichomonas foetus.

The utility of therapeutic vaccination of bulls against Tritrichomonas foetus has been advocated in previous studies, but anecdotal reports suggest this practice does not clear infections and may additionally confound diagnostic testing by reducing parasite burdens below detectable limits. The objective of this study was to characterize the systemic humoral immune response to therapeutic vaccination in T. foetus-infected bulls over a period of four months using an indirect ELISA and to compare the dynamics of this response to culture and PCR results to establish the existence of a relationship (or lack thereof) between immunization and infection status. A study population of 4- to 6-year-old T. foetus-infected beef bulls (n = 20) was divided equally into a treatment group and a control group. The treatment group received two doses of commercially prepared whole cell killed vaccine 2 weeks apart while the control group received injections of vaccine diluent. Blood samples were collected at each injection and at 4 subsequent dates every 4 weeks thereafter (i.e. 0, 2, 6, 10, 14, and 18 wks) to measure IgG1 and IgG2 antibody subisotype response via an indirect ELISA. Preputial smegma samples were collected at the four monthly intervals following vaccination for diagnosis of infection via InPouch™ culture, Modified Diamond's Medium (MDM) culture, and PCR. Humoral response for both IgG isotypes from week 2 through week 18 were significantly increased in vaccinates compared to controls. No significant decrease in infection prevalence was detected in the treatment group for any of the diagnostic methods used. The apparent lack of pathogen clearance during a stimulated immune response suggests that therapeutic vaccination may not be a useful T. foetus management practice.

S100 protein in serum as a prognostic marker for cerebral injury in term newborn infants with hypoxic ischemic encephalopathy.

The astroglial protein S100 is an established biochemical marker for CNS injury in the adult. The aim was to investigate whether S100 in serum is a prognostic marker of cerebral injury in term newborn infants with hypoxic ischemic encephalopathy (HIE) after perinatal asphyxia. Serum S100 was measured on postnatal days 1-4 in 62 term infants with birth asphyxia. The infants were classified for HIE and had follow-up for at least 18 mo. Infants with moderate and severe HIE had significantly higher S100 levels on postnatal day 1 (p = 0.031) and day 2 (p = 0.008) than infants with mild or no HIE. The levels of S100 decreased on days 2 and 3 in all infants with HIE. The median S100 level on postnatal day 1 was higher in nine infants who died neonatally and in 10 infants who developed cerebral palsy (CP), compared with 43 infants with no signs of impairment at follow up, 14.0 (0.5-60.0) microg/L, 20.7 (0.2-64.0) microg/L and 5.5 (0.7-120.0) microg/L, respectively. A level of S100 above 12 microg/L the first day of life was significantly more frequent in infants who died or developed CP than in infants with no impairment at follow up (p = 0.02). Increased S100 levels were significantly inversely correlated with perinatal pH in the infants and associated with abnormal CTG at admission to the labor ward. Early determination of serum S100 may reflect the extent of brain damage in infants with HIE after asphyxia.