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Establishing an immunohistochemical approach for semi-quantitative assessment of the presence of immunoglobulin G (IgG) in equine, canine, and feline corneas.Healthy corneas of horses, dogs, and cats, euthanized because of a fatal disease or an unrecoverable trauma unrelated to and without a history of ophthalmic disease were formalin-fixed, paraffin-embedded, and determined to be pathomorphologically healthy by light microscopy. Automated immunohistochemistry was performed using primary antibodies against IgG, biotin-conjugated secondary antibodies, and streptavidin-peroxidase, as well as diaminobenzidine for visualization. After counterstaining with hematoxylin, epithelium, stroma, Descemet´s membrane (DM), and endothelium were semi-quantitatively scored for the presence of IgG on a 4-grade scale (0 = no, 1 = faint, 2 = medium, 3 = strong staining) by light microscopy.Corneal specimens of 20 horses (40 eyes) with a median age of 15.5 years (range 2-31 years), 12 dogs (21 eyes) with a median age of 10.0 years (range 4-16), and 13 cats (24 eyes) with a median age of 10.0 years (range 2-18) were included in the study. Different sexes and breeds were represented. In all corneas (100%), significant medium signal intensity in the stroma was observed. Variable immunosignal was obtained in epithelium, DM, and endothelium.This method reproducibly allows for the detection of IgG in healthy equine, canine, and feline corneas, particularly stroma. Semi-quantitative results evidence medium presence of IgG in the corneal stroma. Further research is needed to evaluate IgG presence in diseased corneas.

OBJECTIVE To evaluate the effect and safety of topical anti-human vascular endothelial growth factor bevacizumab in dogs with persistent corneal vascularization. ANIMALS STUDIED Prospective case series of 15 adult dogs (20 eyes). PROCEDURES Dogs received 0.25% bevacizumab eye drops BID for 28 days. Follow-ups were scheduled 28 days and 6-7 months after treatment start. Macroscopic findings were scored for conjunctival hyperemia, chemosis, ocular discharge, corneal edema, vascularization, and pigmentation. Vascularized area was assessed by analyzing photographs using an imaging software. RESULTS The treatment response was variable. Some cases showed a marked reduction in vascularized area and edema, while other eyes had subtle signs of improvement. Vascularization score decreased from 1.5 to 1.1 and vascularized area was reduced by 48.8% after 28 days. A thinning of vessels, consolidation of areal bleedings into fine vascular networks, decrease in distal vessel branching, and a change from blurry vascularized beds into demarcated thin vessels were observed. One dog developed a SCCED 6 months after the last bevacizumab administration. Two dogs died 4 and 4.5 months after the last bevacizumab administration, aged 16 and 12 years, respectively. In all events, a causal relationship is unlikely but cannot be ruled out with certainty. CONCLUSIONS Our findings suggest that topical 0.25% bevacizumab may be an effective treatment option for corneal vascularization in dogs. Further long-term placebo-controlled studies with larger patient cohorts are recommended to provide scientific evidence of efficacy and to investigate dosage, safety, possible use as a single treatment, and routes of administration.

Objective Transplantation of minor salivary glands (MSGs) to the conjunctiva is a treatment option for patients suffering from dry eye disease. As there is not enough information about labial and buccal MSGs in dogs, the aim of this study was to provide evidence of the presence of these glands and to investigate their spatial arrangement and excretory ducts. Methods The oral mucosa of the lower lip of 4 dogs and the whole lower jaw of 1 dog were used for histological and microCT analysis. Presence, number, volumes and the tissue depth of MSGs were assessed. Results Histological analysis showed that compact tubulo‐acinar glands were located in the submucosal connective tissue. MicroCT images revealed that 9 to 21 MSGs were arranged in a single row at the level of the dental alveolae. The volume of the MSGs increased from rostral to caudal and the total volume of glandular tissue per animal ranged from 35.01 mm3 to 549.43 mm3. The mean tissue depth of MSGs ranged from 0.57 mm to 1.37 mm (upper surface of glands) and between 1.43 mm and 3.09 mm (lower surface of the glands). Excretory ducts left the dorsal part of the glands and ran in dorso‐rostral direction. Conclusions The location, number and volume of the labial and buccal MSGs in the dog could be detected and described using microCT scans and histology. The present results can provide valuable information for future transplantation of labial MSGs as therapeutic measure against keratoconjunctivitis sicca.

Objective To determine efficacy of contrast‐enhanced ultrasonography (CEUS) using different sulfur hexafluoride (SF6) doses to assess blood flow and perfusion in equine eyes and to evaluate safety of SF6 in horses. Procedures Ocular B‐mode and contrast‐enhanced ultrasonography were performed bilaterally in nine sedated university‐owned horses. Intravenous SonoVue® bolus injections of 5, 10, 15, 20, 25, and 30 mL were administered for 2/18, 5/18, 6/18, 3/18, 1/18, and 1/18 eyes, respectively. Doses were increased based on ascending bodyweight. Each eye within one horse was examined utilizing a different dose. Qualitative blood flow and quantitative perfusion were analyzed. Heart and respiratory rates were monitored nonsedated, sedated, and during first and second minutes of CEUS. Results Qualitative contrast enhancement (CE) was visible in 7/9 animals. Quantitative CE was measurable bilaterally in four horses, unilaterally in three individuals, and not detected in two animals. In all horses with unilateral CE, the positive eye received the higher dose. Fifteen mL dose resulted in significantly shorter time to peak than 10 mL (P

Objective To assess quantitative perfusion of intra‐ and extraocular regions of interest (ROIs) in conscious, healthy dogs utilizing contrast‐enhanced ultrasonography (CEUS); to compare varying enhancement with the first and second bolus injection and in the right and left eye; and to determine the most appropriate examination time. Procedures Gray scale ultrasonography and contrast harmonic imaging using sulfur hexafluoride were performed randomly assigned in both eyes in 10 university‐owned beagles. Perfusion parameters including slope time, time to peak (TTP), peak intensity (PI), and area under the curve (AUC) were measured at individually drawn ROIs (retrobulbar cone = ROI 1, choroid‐retina complex = ROI 2, medial = ROI 3, and lateral anterior uvea = ROI 4). Results Time‐intensity curve parameters revealed no significant differences in eyes examined by the first or second bolus injection (P > 0.05) or in the right or left eye (P > 0.05). Pooled data from all eyes were analyzed. Peak intensity of ROI 2 was significantly higher compared to all other ROIs (P 0.05). Ratios relative to different ROI sizes showed fastest enhancement in the retrobulbar cone and most intense perfusion in the anterior uveal regions. The first minute after contrast injection provided the highest diagnostic value. Conclusion Quantitative perfusion in nondiseased canine eyes revealed consistent parameters. Application of standardized CEUS protocols may be a promising diagnostic tool to differentiate ocular lesions.

Objective To evaluate ocular and general safety of topical anti‐human VEGF bevacizumab and the effect on serum vascular endothelial growth factor (VEGF) values in healthy dogs. Procedures Nine university‐owned beagles received 0.05 mL of 0.25% bevacizumab eyedrops (Avastin®, Roche) in one eye and 0.05 mL of 0.9% saline solution in the other eye as a control, administered at 12 hours intervals over a period of 28 days. Continuous monitoring for vital parameters and ocular examinations were conducted. Complete blood counts including hematology and coagulation parameters were performed before trial start as well as 24 hours, 7 days, and 28 days after trial start. Measurements of serum VEGF values were obtained using an ELISA‐based approach at days 0, 7, and 28. The experiment was designed as a masked placebo‐controlled study. Results No clinical signs of ocular toxicity or systemic incompatibility were noted in any dog at any time point of the study. No signs of pain were present in any dog at any time point. All blood count values remained in normal clinical ranges without relevant variation. There was no significant change in mean serum VEGF values between day 0 and day 7 and between day 0 and day 28. Conclusions The results indicate that topical bevacizumab treatment is safe in healthy dogs. However, further studies are needed to assess safety and efficacy in diseased dogs with naturally occurring corneal neovascularization.

Objective The morphology of the corneal epithelium in two age groups of horses is described. Distribution patterns of proliferation‐, differentiation‐, stem cell‐associated markers and cell junction proteins were assessed. Methods Corneal samples from 12 horses (six foals and six adult horses) were analyzed after H&E staining and immunohistochemistry using the following antibodies: E‐cadherin, β‐catenin, Connexin 43 (Cx43), tight junction protein 1 (TJP1), cytokeratin (CK) 14, CK 19, CK 3, CK 10, vimentin, Ki67, p63, nerve growth factor (NGF), ABCG2, and epithelial growth factor receptor. Semiquantitative analysis of crypt, limbal, peripheral, and central zone was performed. Semithin and ultrathin sections were used for ultrastructural evaluation of the epithelium. Results The height of the epithelium varied between age groups and crypts were consistently present. In the peripheral and central epithelium, three types of basal cells resembling a pseudostratified epithelium were characterized. Potential stem cell markers (CK 14, p63, NGF, and ABCG2) were present in all zones with decreasing frequency toward the center. Cornea‐specific differentiation marker CK 3 was not expressed in the most basal cell layer of the limbal epithelium. E‐cadherin, β‐catenin, and Cx43 revealed a similar apico‐lateral signal pattern throughout the entire epithelium; only TJP1 was additionally seen at the basal surface. Conclusions This study presents a systematic semiquantitative evaluation of the equine corneal epithelium, showing the presence of crypts as potential stem cell niche with CK 14, p63, NGF, and ABCG2 as relevant markers for cells with regenerative capacity. The pseudostratified arrangement of the basal layer was a unique finding.

Objective Impression cytology is a noninvasive investigation of the ocular surface. It uses the adhesive features of different filter papers to collect a monolayer of epithelial cells from the cornea and/or conjunctiva. Samples obtained by impression cytology exhibit all characteristics of an ideal cytology specimen. The aim of this study was to test the feasibility of impression cytology and determine the most appropriate filter paper to achieve maximum diagnostic value of the feline eye. Animals Studied Ten healthy cats. Procedures The study was conducted in two phases. In the first phase, eight different filter papers (FPs) with various pore sizes were tested: 3.0-, 1.2-, 0.8-, 0.45-, 0.22-, 0.05- and 0.025-μm cellulose acetate papers and a 0.4-μm Biopore membrane (BM). Samples were obtained from the superior bulbar and from the inferior palpebral conjunctiva. In the second phase, three different sampling methods – with and without topical anesthesia, and with topical anesthesia and drying of the conjunctiva – were compared employing the BM encased in the intended BM device (BMD). Samples were evaluated for cellularity and quality of cells. Results In the first phase, samples obtained from the superior bulbar conjunctiva with the BM had the most sufficient cellularity and quality. In the second phase, BMD with topical anesthesia and additional drying of the conjunctiva was the most ideal method. Conclusion The BMD may prove to be a suitable diagnostic tool for clinicians. Sampling is quick, processing is simple, and a large area of intact cells can be harvested.

Bilateral phacoclastic uveitis caused by lenticular infection with Encephalitozoon cuniculi is described in a snow leopard. The diagnosis was made on histopathological and immunohistological examination of both eyes submitted after postmortem examination. There was a positive antibody titer for E. cuniculi (1:320). Polymerase chain reaction (PCR) and sequence analysis of formalin-fixed, paraffin-embedded ocular tissue detected the DNA of E. cuniculi, strain III. No other systemic lesions attributable to the E. cuniculi infection were identified.

Objective Guinea pigs have a very low threshold of corneal sensitivity and at the same time nearly no reflex tearing compared to dogs, cats, and horses. The question arose whether there is a general correlation between corneal sensitivity and the quantity of reflex tearing. Animals studied Totally 160 animals of 8 different species (20 animals per species) were investigated. Procedures The corneal touch threshold (CTT) was measured with a Cochet–Bonnet esthesiometer. The palpebral fissure length (PFL) was measured with a calliper ruler. The Schirmer tear test (STT) was modified by adapting the width of the STT strip to the PFL of every species. For the STT II, 0.4% oxybuprocaine was applied. Results Corneal touch threshold: Cows (1.67 g/mm2), horses (1.23 g/mm2), sheep (1.13 g/mm2), goats (1.44 g/mm2), dogs (2.16 g/mm2), and cats (1.33 g/mm2) show similar CTT values. In contrast, rabbits (6.21 g/mm2) and guinea pigs (7.75 g/mm2) show a significantly lower CTT. Tear Production Difference STT I − STT II: Rabbits have the greatest decline in tear production with 38.4%, followed by sheep (33.3%), dogs (31.1%), cats (24.7%), cows (23.7%), horses (18.0%), and goats (14.0%). Guinea pigs have no decline, but a slight increase of −16.0%. Correlation CTT and STT II − STT I Difference: Pearson’s correlation coefficient shows a small, but significant correlation. The coefficient of determination can only forecast a value with 7.1% certainty. Conclusions The high variance and low reproducibility of results suggest that the measuring devices are inappropriate to assess the evaluated parameters. Therefore, no assured correlation between the corneal sensitivity and the quantity of reflex tearing could be found.

Objective To investigate the density and distribution of conjunctival goblet cells (GC) and study the anatomy and microscopic characteristics of glands associated with the eye in chinchillas (Chinchilla Laniger). Procedure 12 chinchillas were included in the study. Conjunctiva (divided into four regions), eyelids, and glands were embedded in paraffin wax, sectioned, stained, and analyzed. Results Highest GC densities were found in the palpebral region of the nasal and temporal conjunctiva of both eyelids (GC index: 25.1–18.2%), and lowest densities, in the bulbar and marginal region of the nasal and temporal conjunctiva of both eyelids (GC index: 1.5–0.0%). Meibomian glands extend along the entire length of both eyelids, and the whole glandular complex broadens toward the temporal canthus. This is macroscopically visible through the conjunctiva. The openings of the Meibomian glands are macroscopically not discernible. The light pink, smooth, and crescent-shaped lacrimal gland lies next to the aforementioned broadened part of the Meibomian glands in the temporal canthus. The whitish, 0.9-cm-long, smooth Harderian gland is firmly attached to the posterior part of the globe and extends nasally from the optic nerve to the equator. Furthermore, chinchillas possess two lacrimal puncta, situated on the inner conjunctival surface of both eyelids near the medial canthus. A pigmented lacrimal canaliculus originates from each punctum. The vestigial nictitating membrane is supported by a hyaline cartilage and is pigmented at its free margin. Conclusions Chinchillas possess a Harderian gland, a lacrimal gland, and Meibomian glands. The GC density in the nasal and temporal palpebral conjunctiva is higher than in guinea pigs.

This report describes three cases of bilateral macrophakia in 3-, 5-, and 9-year-old cats, respectively. All cats were presented because of visual deficits. Ophthalmic examination revealed macrophakia (the vitreous was replaced by the lens) and retinal changes (tapetal hyper-reflectivity, attenuation of retinal vessels, and retinal folds) in all cats. In addition, bilateral subconjunctival orbital fat prolapse in one cat and microphthalmos in another cat were present. To confirm the ophthalmologic diagnosis of macrophakia, gross pathology examination in one cat and ultrasound examination in another cat were performed.

Purpose Identification of Encephalitozoon cuniculi (E. cuniculi) as a possible causative agent for cataracts and uveitis in cats. Methods Within a 12-month study period, cats that were presented with focal anterior cortical or mature cataract and secondary uveitis underwent a complete ophthalmic examination, complete blood count, serum biochemistry, serologic tests for E. cuniculi and tests for feline immunodeficiency virus (FIV), feline infectious peritonitis (FIP), feline leukemia virus (FeLV) and Toxoplasma gondii (T. gondii). PCR for DNA detection of E. cuniculi and T. gondii as well as cytologic examination of aqueous humor after paracentesis and phacoemulsified lens material were also performed. In addition histopathologic examination of the resected anterior lens capsule and attached lens epithelial cells was performed. Serologic testing for antibodies against E. cuniculi was also performed in 100 ophthalmologically healthy cats. Results Eleven (19 eyes) European shorthair cats with a median age of 3.5 years were included. Nine of 11 cats had bilateral cataracts, with 12/19 eyes having focal anterior cortical cataracts and 7/19 eyes having mature cataracts. In 14/19 eyes anterior uveitis was present. All cats had a positive antibody titer (1:80–1:10 000) for E. cuniculi. Encephalitozoon cuniculi DNA was detected by PCR and sequencing in 18/19 lenses and in 10/19 aqueous samples. Five tentative positive results were detected by cytologic examination. Spores were detected in 15/19 samples of lens material with histopathologic staining. Only 2/100 ophthalmologically healthy cats showed a positive antibody titer for E. cuniculi. Conclusion Encephalitozoon cuniculi is a cause of focal anterior cortical cataract and anterior uveitis in cats.

Objective To determine the presence of chlamydial species including recently described chlamydial agents as well as the human pathogen Chlamydophila pneumoniae in feline conjunctivitis. Animal studied Twenty five cats without and 49 cats with conjunctivitis were tested for chlamydia using a Chlamydiaceae real time (RT) PCR (targeting the 23S rRNA gene sequence), a Chlamydiales PCR (targeting the 16S rRNA gene sequence), and cell culture. The PCR products of all positive samples were sequenced and subsequently analyzed using a basic local alignment search tool search. Results Chlamydiaceae RT PCR and subsequent sequence analyses identified C. pneumoniae in five cats in the conjunctivitis group. The presence of Chlamydophila felis was shown in two cats with conjunctivitis. Chlamydiae related to uncultured members of Chlamydiales were detected in three conjunctivitis cases and in one cat without clinical symptoms. Conclusion This study detects for the first time, the known human pathogen C. pneumoniae in feline conjunctivitis cases using Chlamydiaceae RT PCR and sequence analyses.

Objective To investigate the distribution and density of conjunctival goblet cells (GC) and to study the anatomy and microscopic characteristics of glands associated with the eye in the guinea pig. Procedures Twenty-five guinea pigs were used. Meibomian gland openings were counted using biomicroscopy. Conjunctiva, eyelids and glands were embedded in glycol methacrylate and paraffin. Sections were stained with hematoxylin and eosin (H&E), periodic acid Schiff’s reaction (PAS) and Alcian blue (AB). Results Highest GC densities were found in the bulbar and palpebral region of the nasal conjunctiva (GC index: 13.7–16.4%). Lowest GC densities (GC index: 0.0–1.0%) were found in 3/4 limbal regions (nasal and temporal upper eyelid, temporal lower eyelid). Guinea pigs have 27.1 ± 3.0 (mean ± SD) meibomian gland openings in the upper lid and 25.7 ± 2.3 in the lower lid. Difference between upper and lower lid was significant (P = 0.037). Two subconjunctival sebaceous glands occur temporal to each eye. The Harderian gland is very large. In the lacrimal gland three different cell types were distinguished both according to the cell structure and histochemical staining. Conclusions Goblet cell densities are lower in guinea pigs than in dogs and horses. Positive staining with PAS and AB could be an indication that mucins are produced in the lacrimal gland. If so, they may contribute to the mucin layer of the tear film. Both the extraordinarily large Harderian gland and the subconjunctival sebaceous glands produce lipids and may contribute to the lipid layer of the tear film.

Objective A meibometer is a device to measure the delivery rate of lipids on the eyelid margin. The aim of this study is to determine the measuring precision of the new Meibometer® MB550 (Courage-Khazaka electronic GmbH, 50829, Cologne, Germany), linked to a computer, by means of repeated measurements in dogs by different examiners. Procedure Two investigators measured the lipid rate on the eyelid margin in 10 healthy dogs for 10 days. One examiner measured the right eye (OD) and the other measured the left eye (OS) for 5 days. After 5 days, the eyes to be measured were switched between the examiners. The new device was able to record all measurement values as charts and curves in comparison to the previous Meibometer, which displayed only one value. Results Mean meibomium lipid level ± SD in the OD and OS of 10 dogs in 10 days was 211 ± 48 MU and 205 ± 41 MU (meibometer units), respectively. There was no significant difference between OD and OS, but a broad distribution of the values could be found. Gender did not influence the measurements. A significant difference was determined between the measurements in the first 5 days to those in the following 5 days. There was, therefore, also a significant difference in the measurements of the 2 examiners. Conclusion Meibometry is a minimally invasive lipid measuring method on the eyelid margin. Repeated measurement results obtained by two examiners, with the new device Meibometer MB550 linked to a computer, showed a wide range of values. The measuring precision of the new Meibometer MB550 is therefore questionable.

Objective To describe the effects of long-term treatment with four different eye ointment bases (OBs) in cats. Animals studied Ten healthy cats. Procedures The study was performed in two periods. Four different OBs were tested. Hundred grams of OB contained the following: OB-A: 35.17 g liquid paraffin (lp), 64.83 g white petrolatum (wp); OB-B: 10.03 g lp, 84.95 g wp 5.02 g lanolin; OB-C: 18.34 g lp, 51.40 g wp, 25.00 mg KH2PO4, 57.00 mg K2HPO4, 18.90 g eucerinum anhydricum, 11.28 g water for injections; and OB-D: 70 g unguentum lanalcoli, 20 g lp, 10 g aqua conservans. One eye was treated, and the other served as a negative control. Cats received the OBs TID for 28 days. The two study periods were separated by a 4-month washout phase. Samples for conjunctival impression cytology, swabs for bacteriologic and mycologic examination, and cytobrush samples for FHV-1 and Chlamydophila felis PCR detection were obtained. Both eyes were examined daily. Severity of ocular symptoms was scored using a modified Draize eye irritation test. A total of five eyes were treated with OB-A, five with OB-B, four with OB-C, and five with OB-D. Results Treated eyes had significantly higher clinical scores. Eyes receiving OB-A had the highest overall clinical score. The results of bacteriologic and mycologic examination concur with the previously published data. All samples tested were negative for FHV-1 and Chlamydophila felis. There was no significant difference between treated and control eyes upon cytological examination. Conclusion The application of OBs resulted in clinical symptoms in treated eyes. The long-term use of ointments is not well tolerated in cats and may lead to ocular irritation.

A 22-month-old, female rabbit was presented with a 1-day history of acute unilateral exophthalmos. Ultrasonography and computed tomography (CT) of the orbit revealed an orbital mass. Retrobulbar lymphoma was diagnosed following fine-needle aspiration biopsy (FNAB). Thoracic radiographs were normal, and ultrasonography of the abdomen showed focal hypoechoic thickening of the bowel wall and hypoechoic enlarged lymph nodes. The rabbit was euthanized and histopathology identified the retrobulbar mass as B-cell malignant lymphoma of the Harder's gland. Mesenteric lymph nodes, caecum, and both kidneys were also affected. This is the first documented case of malignant lymphoma of the Harder's gland in a rabbit.

Objective Pimecrolimus is an ascomycin derivative that interferes selectively with the activation of T cells and mast cells and inhibits the production of inflammatory cytokines. This study evaluated the efficacy of an experimental ophthalmic formulation of pimecrolimus in treating keratoconjunctivitis sicca (KCS) and chronic superficial keratitis (CSK) in dogs. Animals and procedures Eight dogs with KCS and six with CSK were included. The dogs were of various breeds, suffered from chronic conditions, and had been pretreated unsuccessfully. The affected eyes were treated with 1 drop of an experimental, corn oil-based pimecrolimus 1% formulation three times a day. Parameters evaluated included Schirmer tear test (STT), ocular discharge, conjunctival inflammation, corneal inflammatory cell infiltrate and scarring, and comfort level. Results The effect of pimecrolimus 1% was pronounced (increase in STT values to higher than 4 mm/min, no signs of inflammation) or moderate (increase in STT values of 3–4 mm/min, mild signs of corneal/conjunctival inflammation) in a total of 6/8 animals with KCS. In 4/6 animals with CSK, the effect was either pronounced (total regression of fibrovascular infiltration into the cornea, no corneal scarring) or moderate (distinct regression of pannus, mild corneal scarring). The response to treatment was unsatisfactory in four of 14 animals. Conclusion Results of this exploratory study suggest that topical 1% pimecrolimus may be a new effective treatment for keratoconjunctivitis sicca and chronic superficial keratitis in dogs.

Samples were collected from 36 cats with feline herpesvirus (FHV-1)-related ocular disease (conjunctivitis, epithelial or stromal keratitis, or corneal sequestration), and 17 cats without ocular changes. Corneoconjunctival swabs, scrapings and biopsies were tested in various combinations for presence of FHV-1 DNA using single round (sr) polymerase chain reaction (PCR) and nested PCR (nPCR). Additional swabs from the inferior conjunctival fornix were tested by enzyme-linked immunosorbent assay for Chlamydophila felis antigen. Cytologic evaluation was carried out on conjunctival (cats with conjunctivitis) and corneal (cats with keratitis) cytobrush preparations. FHV-1 DNA was detected by PCR in 14 (39%) cats with ocular disease and 1 (6%) of the control group. Agreement between srPCR and nPCR results was significant (P < 0.01). FHV-1 DNA was detected in 3/7 cats with conjunctivitis, 5/6 cats with epithelial keratitis, 3/11 cats with stromal keratitis, and 3/12 cats with corneal sequestration. There was a significant association (P = 0.0027) between viral presence and epithelial keratitis. However, no significant association was found between viral presence and conjunctivitis (P = 0.059), stromal keratitis (P = 0.15), or corneal sequestration (P = 0.18). With respect to FHV-1 DNA detection, intersample agreement was significant (P < 0.03). No sampling technique seemed more likely than another to harvest detectable viral DNA, except for cats with corneal sequestrum in which viral DNA was not detected using corneoconjunctival swabs. FHV-1 DNA was detected in 6/9 samples with intranuclear inclusion bodies and in 6/7 cats with eosinophils on cytologic examination. All samples tested negative for C. felis antigen.

A 3-year-old spayed female Siberian Husky was presented due to acute vision loss. Examination revealed bilateral optic neuritis and lymphocytic meningoencephalitis. The serum (1:800) and cerebrospinal fluid (CSF; 1:200) immunoglobulin (Ig)G titers for tick-borne encephalitis virus (TBEV) were elevated as were the serum IgG titer for Anaplasma phagocytophilum (1:640) and serum IgM titer for Toxoplasma gondii (1:20). Intracytoplasmic inclusion bodies such as ehrlichial or anaplasmal morulae were not observed in the CSF or blood smear. The dog was treated with methylprednisone and doxycycline. The left eye regained vision; the right eye remained blind. Anti-inflammatory therapy was stopped on day 18 after diagnosis. Four days later the dog showed evidence of hyperesthesia in the cervical region. Analysis of CSF showed no abnormalities and CSF IgG titers for TBEV and A. phagocytophilum were negative. Funduscopic evidence of active papillitis was absent on day 22 in the left eye and on day 86 in the right eye. On day 243, the dog was presented again with lethargy, ataxia, disorientation and temporary head tilt. The IgG titer for TBEV was again elevated in the CSF (1:800) and in serum (1:400). After interpretation of all findings, we assume that meningoencephalitis and optic neuritis in this patient was caused by TBEV and associated immune-mediated inflammation. In endemic areas, TBEV should be considered as cause of optic neuritis in dogs.

To investigate the density and distribution of conjunctival goblet cells (GC) and study the anatomy and microscopic characteristics of glands associated with the eye in chinchillas (Chinchilla Laniger).12 chinchillas were included in the study. Conjunctiva (divided into four regions), eyelids, and glands were embedded in paraffin wax, sectioned, stained, and analyzed.Highest GC densities were found in the palpebral region of the nasal and temporal conjunctiva of both eyelids (GC index: 25.1-18.2%), and lowest densities, in the bulbar and marginal region of the nasal and temporal conjunctiva of both eyelids (GC index: 1.5-0.0%). Meibomian glands extend along the entire length of both eyelids, and the whole glandular complex broadens toward the temporal canthus. This is macroscopically visible through the conjunctiva. The openings of the Meibomian glands are macroscopically not discernible. The light pink, smooth, and crescent-shaped lacrimal gland lies next to the aforementioned broadened part of the Meibomian glands in the temporal canthus. The whitish, 0.9-cm-long, smooth Harderian gland is firmly attached to the posterior part of the globe and extends nasally from the optic nerve to the equator. Furthermore, chinchillas possess two lacrimal puncta, situated on the inner conjunctival surface of both eyelids near the medial canthus. A pigmented lacrimal canaliculus originates from each punctum. The vestigial nictitating membrane is supported by a hyaline cartilage and is pigmented at its free margin.Chinchillas possess a Harderian gland, a lacrimal gland, and Meibomian glands. The GC density in the nasal and temporal palpebral conjunctiva is higher than in guinea pigs.

Identification of Encephalitozoon cuniculi (E. cuniculi) as a possible causative agent for cataracts and uveitis in cats.Within a 12-month study period, cats that were presented with focal anterior cortical or mature cataract and secondary uveitis underwent a complete ophthalmic examination, complete blood count, serum biochemistry, serologic tests for E. cuniculi and tests for feline immunodeficiency virus (FIV), feline infectious peritonitis (FIP), feline leukemia virus (FeLV) and Toxoplasma gondii (T. gondii). PCR for DNA detection of E. cuniculi and T. gondii as well as cytologic examination of aqueous humor after paracentesis and phacoemulsified lens material were also performed. In addition histopathologic examination of the resected anterior lens capsule and attached lens epithelial cells was performed. Serologic testing for antibodies against E. cuniculi was also performed in 100 ophthalmologically healthy cats.Eleven (19 eyes) European shorthair cats with a median age of 3.5 years were included. Nine of 11 cats had bilateral cataracts, with 12/19 eyes having focal anterior cortical cataracts and 7/19 eyes having mature cataracts. In 14/19 eyes anterior uveitis was present. All cats had a positive antibody titer (1:80-1:10,000) for E. cuniculi. Encephalitozoon cuniculi DNA was detected by PCR and sequencing in 18/19 lenses and in 10/19 aqueous samples. Five tentative positive results were detected by cytologic examination. Spores were detected in 15/19 samples of lens material with histopathologic staining. Only 2/100 ophthalmologically healthy cats showed a positive antibody titer for E. cuniculi.Encephalitozoon cuniculi is a cause of focal anterior cortical cataract and anterior uveitis in cats.

To determine the presence of chlamydial species including recently described chlamydial agents as well as the human pathogen Chlamydophila pneumoniae in feline conjunctivitis.Twenty five cats without and 49 cats with conjunctivitis were tested for chlamydia using a Chlamydiaceae real time (RT) PCR (targeting the 23S rRNA gene sequence), a Chlamydiales PCR (targeting the 16S rRNA gene sequence), and cell culture. The PCR products of all positive samples were sequenced and subsequently analyzed using a basic local alignment search tool search.Chlamydiaceae RT PCR and subsequent sequence analyses identified C. pneumoniae in five cats in the conjunctivitis group. The presence of Chlamydophila felis was shown in two cats with conjunctivitis. Chlamydiae related to uncultured members of Chlamydiales were detected in three conjunctivitis cases and in one cat without clinical symptoms.This study detects for the first time, the known human pathogen C. pneumoniae in feline conjunctivitis cases using Chlamydiaceae RT PCR and sequence analyses.

To investigate the distribution and density of conjunctival goblet cells (GC) and to study the anatomy and microscopic characteristics of glands associated with the eye in the guinea pig.Twenty-five guinea pigs were used. Meibomian gland openings were counted using biomicroscopy. Conjunctiva, eyelids and glands were embedded in glycol methacrylate and paraffin. Sections were stained with hematoxylin and eosin (HE), periodic acid Schiff's reaction (PAS) and Alcian blue (AB).Highest GC densities were found in the bulbar and palpebral region of the nasal conjunctiva (GC index: 13.7-16.4%). Lowest GC densities (GC index: 0.0-1.0%) were found in 3/4 limbal regions (nasal and temporal upper eyelid, temporal lower eyelid). Guinea pigs have 27.1±3.0 (mean±SD) meibomian gland openings in the upper lid and 25.7±2.3 in the lower lid. Difference between upper and lower lid was significant (P=0.037). Two subconjunctival sebaceous glands occur temporal to each eye. The Harderian gland is very large. In the lacrimal gland three different cell types were distinguished both according to the cell structure and histochemical staining.Goblet cell densities are lower in guinea pigs than in dogs and horses. Positive staining with PAS and AB could be an indication that mucins are produced in the lacrimal gland. If so, they may contribute to the mucin layer of the tear film. Both the extraordinarily large Harderian gland and the subconjunctival sebaceous glands produce lipids and may contribute to the lipid layer of the tear film.

Objective To establish reference values for Schirmer tear tests (STT) I and II, phenol red thread (PRT) tear test and eye blink frequency, and to determine corneal sensitivity for normal guinea pigs. Animals studied One hundred and eight eyes of 54 adult Duncan-Hartley guinea pigs. Procedure Schirmer tear test (STT) I and then STT II were performed in 36 guinea pigs. PRT and STT I were compared in 18 adult Duncan-Hartley guinea pigs. Corneal sensitivity was determined in 23 guinea pigs by evaluating the corneal touch threshold (CTT) of five different regions using a Cochet-Bonnet esthesiometer. Eye blink frequency was measured in 10 guinea pigs over a period of 20 min and in 17 guinea pigs over a period of 10 min. Results Mean STT I was 0.36 mm ± 1.09 mm (wetting/min) and mean STT II was 0.43 mm ± 1.29 mm (wetting/min). There was no significant difference between mean STT I and mean STT II (P = 0.79). The mean PRT-value was 16 ± 4.7 mm (wetting/15 s), and the mean STT I-value in the same guinea pigs was 0.6 ± 1.83 mm (wetting/min). Corneal sensitivity was significantly higher in the center than in the four limbal regions. The mean CTT for central, ventral, nasal, temporal and dorsal regions was 2, 1.7, 1.7, 1.7 and 1.6 cm or 3.7, 5.2, 5.6, 5.7 and 6.4 g/mm2, respectively. Eye blink frequency was between two to five (mean 3.4 ± 1.04) blinks per eye over 20 min in guinea pigs in their home environment, while in handheld and restrained guinea pigs eye blink frequency showed a variation between 0 and 17 blinks per eye (mean 3.24 ± 3.64 blinks per eye) over 10 min. Conclusions As there were no significant differences between STT I and STT II results, reflex tear secretion in the guinea pig may not exist. The most likely explanation is a lower corneal sensitivity in the guinea pig than in other species, such as cats, dogs and horses. Because of the small amount of tears, PRT is the preferred test for tear measurement in the guinea pig.

To determine the effect of various mydriatics (1% atropine, 1% cyclopentolate, 0.5% tropicamide, 10% phenylephrine) on intraocular pressure (IOP) and pupil size (PS) in normal cats.The mydriatics were tested in 10 adult ophthalmoscopically normal European Domestic Short-haired cats. Procedure Single-dose drug studies were divided into placebo (vehicle of phenylephrine), 10% phenylephrine, 0.5% tropicamide, 1% cyclopentolate and 1% atropine. After measurement of IOP and pupil size (PS) at 8 a.m. on the first day, one drop of the tested drug was applied to one randomly selected eye. The IOP and PS were measured for a minimum of 36 h until the pupil returned to pretest size.Ten per cent phenylephrine had no significant effect on IOP, and the effect on the pupil size was minimal (or= 1 mm difference). One per cent atropine showed a maximal mean difference of 4.3 mmHg +/- 4.1 mmHg in IOP compared to the untreated eye. These differences were statistically significant at 1, 3, 6, 12 and 16 h post-treatment. At 1, 1.5 and 2 h after unilateral application of 1% cyclopentolate, the IOP in the treated eye was significantly higher than the IOP in the untreated eye with a maximal mean difference of 4.1 mmHg +/- 4.5 mmHg. With 0.5% tropicamide, IOP of the treated eye was significantly higher than IOP of the untreated eye at 1 and 1.5 h after treatment with a maximal mean difference of 3.5 mmHg +/- 3.3 mmHg. One per cent atropine caused the greatest increase in IOP, followed by 1% cyclopentolate and 0.5% tropicamide. The dilating effect of atropine, cyclopentolate and tropicamide lasted longer than the effect on intraocular pressure.The parasympatholytics 1% atropine, 1% cyclopentolate and 0.5% tropicamide caused a statistically significant elevation of IOP and produced mydriasis in the treated eye in normal cats, while the sympathomimetic 10% phenylephrine had no significant effect on IOP or pupil size.

Purpose The purpose of this study was to determine if intravenous hypertonic hydroxyethyl starch (7.5%/6%) (HES) could decrease the intraocular pressure (IOP) in healthy normotensive dogs, and compare its effect with that of mannitol (20%) (experimental study). In addition, the potential IOP-lowering effect of hypertonic HES was evaluated in six dogs with primary glaucoma (clinical study). Material and methods Experimental study: eight male ophthalmoscopically and clinically healthy Beagles were included in this study. The IOP of each dog was measured by applanation tonometry in both eyes to obtain control values at 10:00, 10:15, 10:30, 10:45, 11:00 a.m., and then every hour until 6:00 p.m. prior to the first treatment (control period). Each dog received, with at least 2-week intervals and in a random order, an intravenous (IV) infusion of 4 mL/kg hypertonic HES (1.2 g/kg NaCl; 0.96 g/kg HES) and 4 mL/kg mannitol 20% (1 g/kg) over a period of 15 min starting at 10:00 a.m. IOP was measured oculus uterque (OU) at the same time intervals as in the control study. The differences in IOP between the treatment groups and the baseline IOP (before the start of infusion), between oculus sinister (OS) and oculus dexter (OD) and between the same time points of all groups were determined with a Student's t-test for paired samples (P = 0.05). Clinical study: six dogs with primary glaucoma (representing seven eyes) received an IV infusion of 4 mL/kg hypertonic HES over a period of 15 min. IOP was measured before and 15 and 30 min after starting the infusion. Results Experimental study: no significant difference between IOP of both eyes was found. A significant decrease in IOP from baseline value was recorded at 15, 30, 45, and 60 min after the start of mannitol infusion (mean amplitude in IOP decrease 3.21 mmHg; P

A 7-year-old female spayed European shorthair cat was presented because of visual reduction and a green reflex in the right eye. Examination revealed a 'D-shaped' pupil in the right eye and a 'reverse-D-shaped' pupil in the left eye. Both eyes showed fibrinous anterior uveitis with exudates in the anterior chamber and precipitates on the posterior surface of the cornea. In both eyes a chorioretinitis was diagnosed which caused partial retinal detachment and peripapillary oedema in the left eye. Histopathology revealed intestinal and ocular lymphosarcoma. The aberrant pupil shapes which persisted after death were due to infiltration of the iris stroma and were not caused neurologically. The observation that in both eyes only the medial parts of the iris were affected could not be explained.