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Immune-mediated ocular inflammation is a common clinical diagnosis reached for horses with keratitis and uveitis. This diagnosis is made as a diagnosis of exclusion following a thorough effort to rule out an underlying cause for the inflammation, most importantly infectious and neoplastic disease. Practically, response to ophthalmic and systemic anti-inflammatory or immunomodulatory medications is used to support a diagnosis of immune-mediated ocular inflammation; however, such medications are often contraindicated in the face of infection or neoplasia. This article will summarize our current understanding and approach to the diagnosis and management of immune-mediated keratitis and recurrent or insidious uveitis in horses., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Objective: The objective of this study was to use shotgun label-free tandem mass spectrometry (LF-MS/MS) to evaluate aqueous humor (AH) from horses with uveitis (UH) compared to ophthalmologically healthy horses (HH)., Animals Studied: Twelve horses diagnosed with uveitis based on ophthalmic examination and six ophthalmologically healthy horses (postmortem) purchased for teaching purposes., Procedures: All horses received a complete ophthalmic examination and physical exam. Aqueous paracentesis was performed on all horses and AH total protein concentrations were measured with nanodrop (TPn) and refractometry (TPr). AH samples were analyzed with shotgun LF-MS/MS and proteomic data were compared between groups using Wilcoxon rank-sum test., Results: A total of 147 proteins were detected, 11 proteins had higher abundance in UH, and 38 proteins had lower abundance in UH. Proteins with higher abundance included apolipoprotein E, alpha-2-macroglobulin (A2M), alpha-2-HS-glycoprotein, prothrombin, fibrinogen, complement component 4 (C4), joining chain for IgA and IgM, afamin, and amine oxidase. There were positive correlations between TPn (p = .003) and TPr (p = .0001) compared to flare scores., Conclusion: Differential abundance of A2M, prothrombin, fibrinogen, and C4 indicate upregulation of the complement and coagulation cascade in equine uveitis. Proinflammatory cytokines and the complement cascade have potential as therapeutic targets for equine uveitis., (© 2023 The Authors. Veterinary Ophthalmology published by Wiley Periodicals LLC on behalf of American College of Veterinary Ophthalmologists.)

Regenerative therapy and biologics have the promise to treat equine ocular surface diseases, including corneal ulceration or immune-mediated keratitis, or intraocular diseases such as uveitis. The use of blood-derived products such as serum or platelet-rich plasma, mesenchymal stem cells, or amniotic membrane grafts may be beneficial for the treatment of ulcerative and chronic keratitis in horses. Furthermore, the use of stem cells or gene therapy has promise for the treatment of Intraocular diseases such as equine recurrent uveitis by providing efficacious, practical, and long-term therapy for these blinding diseases., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Equine recurrent uveitis (ERU) is the leading cause of blindness for horses; previous research implicated the leopard complex spotting allele (LP) as a genetic risk factor for insidious uveitis in the Appaloosa. There is limited information about risk in the Knabstrupper., Objective: To evaluate clinical manifestations, disease frequency and potential risk factors for ERU in Knabstrupper horses., Study Design: Cross-sectional study., Methods: Ocular examinations were performed on 116 horses, and based on identified anomalies, horses were classified as suspect, ERU-affected or having no clinical signs. Microagglutination testing (MAT) of serum assessed exposure to Leptospira spp. Clinical signs, age, sex, base colour, coat pattern, LP and PATN1 genotypes, percent white at birth, progressive roaning and Leptospira were assessed as risk factors using multivariable exact logistic regression, accounting for clustering at the barn level. Additionally, a pedigree analysis was performed (n = 20 cases and 21 controls), and coefficients of coancestry (CC) and inbreeding were calculated., Results: Prevalence of insidious uveitis in this sample of Knabstruppers was 20.7%. Similar to findings for Appaloosas, LP homozygotes had higher odds of uveitis compared with true solid (N/N) horses (LP/LP OR = 7.64, 95% CI [0.8 to +INF], p = 0.04) and age was also identified as a risk factor. After accounting for LP, the 16-20 age group had higher odds compared with the youngest group (OR = 13.36, 95% CI [1.4-213.4], p = 0.009). The distributions of average CC were significantly different between cases and controls (p = 0.01)., Main Limitations: A relatively small sample size decreased the power for detecting additional associations. The progressive nature of insidious uveitis may have prevented identification of younger affected horses., Conclusions: Our data support genotyping for LP to assess risk of ERU in Knabstruppers. Additional studies are necessary to develop more robust risk models across LP breeds for earlier detection and improved clinical management., (© 2022 EVJ Ltd.)

Objective: The objective of the study was to determine the effect of gentamicin on CD3+ T-lymphocyte proliferation and cell viability using an in vitro cell culture model as a means of investigating the mechanism of action of low-dose intravitreal gentamicin injection., Animals Studied: Three adult horses with no evidence of ophthalmic or systemic disease., Procedure: Peripheral blood lymphocytes were treated with gentamicin at concentrations 37.5 μg/mL, 112.5 μg/mL, 187 μg/mL, 375 μg/mL, or 750 μg/mL then stimulated to proliferate with concanavalin A (ConA). 4',6-diamidino-2-phenylindole (DAPI) and carboxyfluoroscein succinimidyl ester (CSFE) were used as markers of cell viability and cell proliferation, respectively. Following 5-day culture, live cell counts and CSFE fluorescent intensity data were collected via automated cell count and flow cytometry. The experimental design was duplicated using preservative-free gentamicin and a proprietary brand formulation. Statistical analysis was performed using two-way ANOVA with Tukey's multiple comparison test., Results: No statistically significant comparisons in CD3+ T-lymphocyte live cell counts and geometric mean fluorescent intensity of CSFE were identified between gentamicin concentrations or formulations., Conclusions: Gentamicin had no effect on equine peripheral blood CD3+ T-lymphocyte cell viability and proliferation in concentrations ranging from "safe" to "retinotoxic" in relation to intravitreal injection volumes. Low-dose intravitreal gentamicin may not suppress the Th1- and Th17-mediated immune response., (© 2023 The Authors. Veterinary Ophthalmology published by Wiley Periodicals LLC on behalf of American College of Veterinary Ophthalmologists.)

Purpose: To evaluate the correlation between equine pectinate ligament descemetization and ocular disease., Methods: The pathology database of the North Carolina State University Veterinary Medical Center was searched from 2010-2021 for all equine globes. Disease status was then assigned as affected by glaucoma, uveitis, or "other" based upon clinical records. The iridocorneal angles (ICA) of each globe were evaluated for the presence of pectinate ligament descemetization, the length of descemetization, as well as for the degree of angle collapse and the extent of cellular infiltrate or proteinaceous debris. One slide from each eye was evaluated by two separate, blinded investigators (HW & TS)., Results: A total of 66 eyes from 61 horses were identified, with a total of 124 sections of ICA of sufficient quality to review. 16 horses were affected by uveitis, 8 by glaucoma, 7 by both glaucoma and uveitis, and 30 horses by other ocular disease, most commonly ocular surface disease or neoplasia, which served as controls. Pectinate ligament descemetization was most prevalent in the control group compared to the glaucoma and uveitis groups. Pectinate ligament descemetization length was positively correlated with age, with an increase of 13.5 μm per year of age (p = .016). Infiltrate scores and angle closure scores were higher in both the glaucoma and uveitis group compared to the control group (p < .001)., Conclusions: Equine pectinate ligament descemetization appears to be correlated with increased age and should not be used as a histologic marker for the presence of glaucoma., (© 2023 The Authors. Veterinary Ophthalmology published by Wiley Periodicals LLC on behalf of American College of Veterinary Ophthalmologists.)

Horses with recurrent uveitis can be treated by intravitreal injection of low dose gentamicin under sedation and after local anesthetic techniques including the retrobulbar nerve block. Since it is reported that retinal degeneration can be secondary to an acute increase of intraocular pressure (IOP), the current randomized controlled study was carried out in order to investigate the changes in IOP following retrobulbar anesthesia, with two different volumes of local anesthetic (lidocaine) solution (10 and 5 mL), and intravitreal injection of 6 mg gentamicin in two different volumes of solution (0.3 and 0.15 mL). Multivariate regression showed that IOP was significantly lower in the pathologic eye at baseline (estimated effect size -6.1 mmHg; P = .0001) and after sedation (estimated effect size -9.4 mmHg; P < .0001). The IOP was significantly higher after a 10 mL block (P .0002) but this effect was very small (+0.5 mmHg). There was no significant difference in IOP after the injection of 0.15 or 0.3 mL solution of gentamicin. There was no significant combined effect of the volume of local anesthetic used for the block and the volume of the gentamicin solution. Since the expected anesthetic effects (desensitization and akinesia) were met in all cases, the smaller volume of 5 mL of lidocaine solution would be preferable for retrobulbar block for intravitreal injections, while intravitreal injections volumes of 0.15 and 0.30 mL can be used indifferently., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Objective: To determine the prevalence of ocular disease in draft horses in the United States., Animals: Draft horses of various breeds and ages., Procedure: Nondilated ophthalmic examination was performed using slit lamp biomicroscopy and indirect ophthalmoscopy. Intraocular pressures were measured when possible., Results: One hundred sixty-five draft horses were examined. Age range: 10 days to 33 years (mean 10.8 years, median 10 years); 87 geldings (52.7%), 71 mares (43.0%), 7 stallions (4.2%); 64 Percherons (38.8%), 51 Belgians (30.9%), 29 Clydesdales (17.6%), 15 Shires (9%), and 6 other draft breed (3.6%). Intraocular pressure: mean 24.7 mmHg OD, range 13-37 mmHg; mean 25.0 mmHg OS, range 11-37 mmHg. Vision-threatening disease was present in 9 horses (5.5%): complete cataracts 1, post-traumatic optic nerve atrophy 1, uveitis and secondary glaucoma 1, retinal detachment 1, large chorioretinal scar 3, phthisis bulbi 2. Non-vision-threatening ocular disease was present in 56 horses (33.9%) involving one or more ocular structures: eyelid trauma/notch defect 14 (8.5%), SCC-type adnexal lesions 12 (7.3%), corneal scars 16 (9.7%), keratitis 6 (3.6%), corpora nigra cyst 15 (9.1%), incipient/punctate cataract 50 (30.3%), vitreous degeneration 10 (6.1%), asteroid hyalosis 1, "bullethole" chorioretinal scars 3, RPE coloboma 1. Linear keratopathy was present in 28 horses (17%) with 2/28 having concurrent vision threatening ocular disease., Conclusions: Ocular abnormalities, in particular minor cataracts, were relatively common in this population, but not typically vision-threatening. Additionally, this survey demonstrated a greater prevalence of linear keratopathy in draft horses compared with reports in other breeds; however, it does not appear to be associated with concurrent ocular disease., (© 2022 The Authors. Veterinary Ophthalmology published by Wiley Periodicals LLC on behalf of American College of Veterinary Ophthalmologists.)

Equine recurrent uveitis (ERU) is an ocular inflammatory disease that can be difficult to manage clinically. As such, it is the leading cause of bilateral blindness for horses. ERU is suspected to have a complex autoimmune etiology with both environmental and genetic risk factors contributing to onset and disease progression in some or all cases. Work in recent years has aimed at unraveling the primary triggers, such as infectious agents and inherited breed-specific risk factors, for disease onset, persistence, and progression. This review has aimed at encompassing those factors that have been associated, implicated, or substantiated as contributors to ERU, as well as identifying areas for which additional knowledge is needed to better understand risk for disease onset and progression. A greater understanding of the risk factors for ERU will enable earlier detection and better prognosis through prevention and new therapeutics., (© 2022 American College of Veterinary Ophthalmologists.)

Background: Risk factors for a first episode of primary uveitis in horses have not been determined. In addition, disease progression and the proportion of horses that develop recurrence following the original episode are not known., Objectives: To determine the risk factors for the development of a first episode of primary uveitis in horses in the UK and to document the proportion of cases that experience recurrence following this first episode., Study Design: Prospective case-control longitudinal study., Methods: Horses with a first episode of primary uveitis between July 2014 and August 2018 were recruited to the study. For each case, two controls were selected. A questionnaire was completed for each horse and multivariable logistic regression analysis was carried out to identify associations between horse and management-level variables and the risk of uveitis. Each case was then followed longitudinally to determine the rate of recurrence of uveitis., Results: Twenty-three cases and 46 controls were recruited. Being close to a pig farm (OR 27.8, CI 1.31-592.06) and a recent history of flooding of the pasture (OR 15.43, CI 2.80-84.98) was associated with increased risk of uveitis. Being in the same owner's possession for a longer amount of time had a protective effect (OR 0.79, CI 0.68-0.93). Horses recovered uneventfully following treatment and showed no evidence of recurrence in 59.1% of the cases (n = 13). In five horses (22.7%), the initial episode of uveitis could not be controlled and required surgical therapy. Recurrence was observed in four horses (18.2%)., Main Limitations: Small sample size. Recall bias from owners for some of the data. Possibility of selection bias., Conclusions: Clinicians can use this information to identify horses with increased risk of uveitis and provide advice to the owners of these horses. The proportion of cases that experience recurrence appears low in the UK., (© 2022 EVJ Ltd.)

Equine recurrent uveitis (ERU) is a blinding ocular disorder among horses, and the Appaloosa horse breed is disproportionally affected by a chronic form of this intraocular inflammatory disease known as insidious uveitis. Strong breed predisposition and previous investigations suggest that there is a genetic component to the pathology of insidious uveitis among Appaloosa horses; however, no estimates of the heritability of the disease have previously been determined. This study aimed to characterize the genetic underpinning of the disease by estimating the heritability for insidious uveitis among Appaloosas. After combining two genotyping array datasets from the Illumina Equine SNP70 BeadChip and the Axiom Equine 670 K Genotyping Array, heritability was estimated for 59 affected and 83 unaffected horses using both restricted maximum likelihood (REML) and phenotype correlation - genotype correlation solvers from the linkage disequilibrium adjusted kinship software. Based on previous research, age and sex were used as covariates, and the locus responsible for the characteristic Appaloosa coat pattern (LP), previously associated with ERU risk, was included as a fixed effect ('top predictor'). Using prevalence values from 0.05 to 0.42, the heritability estimate for insidious uveitis ranged from 0.95 (SE = 0.14) to 1.74 (SE = 0.25) with LP contributing 0.16-0.33 to the estimate. This study suggests that insidious uveitis is highly heritable (REML 95% CI, h 2  = 0.68-1.0) and additional loci outside of LP are contributing to the genetic risk for insidious uveitis for Appaloosas. Once identified, these other genetic factors may lead to new disease mitigation efforts in veterinary care and breeding practices., (© 2022 Stichting International Foundation for Animal Genetics.)

This review, which is part of the "Currents in One Health" series, describes the importance of the study of immune-mediated ocular disease in the development of innovative therapeutics, such as cell and gene therapy for the eye. Recent examples of cell and gene therapy studies from the author's laboratory are reviewed to emphasize the importance of One Health initiatives in developing innovative therapies for ocular diseases. Spontaneous immune-mediated corneal disease is common in horses, cats, dogs, and humans. Autologous bone marrow-derived mesenchymal stem cells (BM-MSCs) injected subconjunctivally resulted in the resolution of naturally occurring immune-mediated keratitis (IMMK) without adverse effects. These results support that autologous subconjunctival BM-MSC therapy may be a viable treatment alternative for IMMK. Furthermore, the use of subconjunctival MSCs may be an effective method to treat ocular surface immune-mediated diseases in humans and other species, including herpetic stromal keratitis and immunologic dry eye disease. Furthermore, the use of adeno-associated viral (AAV) vectors to deliver the immunosuppressive transgene cDNA of equine interleukin 10 (eqIL-10) or human leukocyte antigen G injected intravitreally was shown to be safe and inhibited the development of uveitis in the experimental autoimmune uveitis rat model. Efficacy and safety studies of ocular gene therapy in models will pave the way for clinical trials in animals with naturally occurring immune-mediated diseases, such as a therapeutic clinical trial for AAV-eqIL-10 in horses with equine recurrent uveitis.

In this article, which is part of the Currents in One Health series, the role of naturally occurring ocular disease in animals is reviewed with emphasis on how the understanding of these ocular diseases contributes to one health initiatives, particularly the pathogenesis and treatment of ocular diseases common to animals and humans. Animals spontaneously develop ocular diseases that closely mimic those in humans, especially dry eye disease, herpes virus infection (cats), fungal keratitis (horses), bacterial keratoconjunctivitis, uveitis, and glaucoma. Both uveitis and glaucoma are common in domestic animals and humans, and many similarities exist in pathogenesis, genetics, and response to therapy. Furthermore, the study of inherited retinal disease in animals has particularly epitomized the one health concept, specifically the collaborative efforts of multiple disciplines working to attain optimal health for people and animals. Through this study of retinal disease in dogs, innovative therapies such as gene therapy have been developed. A unique opportunity exists to study ocular disease in shared environments to better understand the interplay between the environment, genetics, and ocular disease in both animals and humans. The companion Currents in One Health by Gilger, AJVR, December 2022, addresses in more detail recent studies of noninfectious immune-mediated animal ocular disease and their role in advancing ocular health globally.

Background: Due to recent developments in artificial intelligence, deep learning, and smart-device-technology, diagnostic software may be developed which can be executed offline as an app on smartphones using their high-resolution cameras and increasing processing power to directly analyse photos taken on the device., Objectives: A software tool was developed to aid in the diagnosis of equine ophthalmic diseases, especially uveitis., Study Design: Prospective comparison of software and clinical diagnoses., Methods: A deep learning approach for image classification was used to train software by analysing photographs of equine eyes to make a statement on whether the horse was displaying signs of uveitis or other ophthalmic diseases. Four basis networks of different sizes (MobileNetV2, InceptionV3, VGG16, VGG19) with modified top-layers were evaluated. Convolutional Neural Networks (CNN) were trained on 2346 pictures of equine eyes, which were augmented to 9384 images. 261 separate unmodified images were used to evaluate the performance of the trained network., Results: Cross validation showed accuracy of 99.82% on training data and 96.66% on validation data when distinguishing between three categories (uveitis, other ophthalmic diseases, healthy)., Main Limitations: One source of selection bias for the artificial intelligence presumably was the increased pupil size, which was mainly present in horses with ophthalmic diseases due to the use of mydriatics, and was not homogeneously dispersed in all categories of the dataset., Conclusions: Our system for detection of equine uveitis is unique and novel and can differentiate between uveitis and other equine ophthalmic diseases. Its development also serves as a proof-of-concept for image-based detection of ophthalmic diseases in general and as a basis for its further use and expansion., (© 2021 The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.)

Equine recurrent uveitis (ERU) is an autoimmune disease defined by inflammation of the uveal tract of the eye. The cause of ERU is thought to be complex, involving both genetic and environmental factors. The purpose of this study was to investigate potential genetic risk factors for ERU in the Icelandic horse. Fifty-six Icelandic horses (11 affected with ERU and 45 controls) living in Denmark and the USA, eight years or older, were included in the study. A case-control GWAS was performed using the GGP Equine 80K array on the Illumina Infinium HD Beadchip using 40 horses. A mixed linear model analysis identified a single SNP on ECA 11 (BIEC2&#95;141650; NC&#95;009154.3:g.3817009A>G) that reached genome-wide significance (p = 1.79 × 10 -7 ). This variant was within an intron of tissue inhibitor of metalloproteinase 2 (TIMP2), a gene previously implicated in ERU. Sanger sequencing identified a single coding variant in this gene; however it was a synonymous mutation (NC&#95;009154.3:g.3858193C>T) and was not perfectly concordant with ERU phenotype (p = 0.68). Further investigation of TIMP2 is warranted. Additional horses and markers are needed to identify other potential loci worthy of further investigation as contributors to ERU risk in Icelandic horses., (© 2022 Stichting International Foundation for Animal Genetics.)

Equine recurrent uveitis (ERU) is a painful and debilitating autoimmune disease and represents the only spontaneous model of human recurrent uveitis (RU). Despite the efficacy of existing treatments, RU remains a leading cause of visual handicap in horses and humans. Cytokines, which utilize Janus kinase 2 (Jak2) for signaling, drive the inflammatory processes in ERU that promote blindness. Notably, suppressor of cytokine signaling 1 (SOCS1), which naturally limits the activation of Jak2 through binding interactions, is often deficient in autoimmune disease patients. Significantly, we previously showed that topical administration of a SOCS1 peptide mimic (SOCS1-KIR) mitigated induced rodent uveitis. In this pilot study, we test the potential to translate the therapeutic efficacy observed in experimental rodent uveitis to equine patient disease. Through bioinformatics and peptide binding assays we demonstrate putative binding of the SOCS1-KIR peptide to equine Jak2. We also show that topical, or intravitreal injection of SOCS1-KIR was well tolerated within the equine eye through physical and ophthalmic examinations. Finally, we show that topical SOCS1-KIR administration was associated with significant clinical ERU improvement. Together, these results provide a scientific rationale, and supporting experimental evidence for the therapeutic use of a SOCS1 mimetic peptide in RU., (© 2022. The Author(s).)

Purpose: To describe the most common ocular abnormalities in the Icelandic horse with focus on equine recurrent uveitis (ERU) and association between ocular abnormalities and summer eczema and coat colors., Methods: A descriptive cross-sectional study. A complete physical and ophthalmic examination as well as measurement of serum titers for Leptospira serovariants was performed on Icelandic horses from Denmark (DK) and the United States (USA)., Results: One hundred and twelve Icelandic horses living in DK and 26 Icelandic horses living in the United States were included in this study (total of 138 horses, 274 eyes). The three most common ocular abnormalities were follicular conjunctivitis (55.8%, 153/274 eyes), cataracts (27.4%, 75/274 eyes), and multifocal chorioretinopathy ("bullet holes"; 19%, 52/274 eyes). The prevalence for ERU among Icelandic horses aged "eight years and older" was 8% (6/75 horses). The "insidious ERU" type was found in 83% of the ERU cohort (5/6 horses), while "classic ERU" was found in 17% (1/6 horses). A correlation between ERU and cataracts was found in the population (p < .01). Leptospiral serology results were not associated with ERU in either DK or US-Icelandic horses (p = 1.00, p = 1.00, respectively). ERU was not associated with summer eczema (p = .49), and no coat colors were associated with the ocular abnormalities found in the Icelandic horse (all p-values > .05)., Conclusion: Follicular conjunctivitis, cataracts, and multifocal chorioretinopathy were the three most common ocular abnormalities findings. Icelandic horses who were 8 years or older had an 8% prevalence for ERU. Summer eczema and coat color were not associated with evidence of ERU or other ocular abnormalities., (© 2021 American College of Veterinary Ophthalmologists.)

Equine recurrent uveitis (ERU) is a common ocular disease of horses and described as a model for human autoimmune uveitis. This immune-mediated, inflammatory condition progressively destroys the eye, ultimately leading to blindness. Genetic and autoimmune factors, next to infections with Leptospira , are discussed as key factors in the pathogenesis. Furthermore, a release of neutrophil extracellular traps (NETs) by activated neutrophils is involved. NETs are composed of decondensed chromatin and proteins that can immobilize invading pathogens. However, if NETs accumulate, they can contribute to detrimental autoimmune processes. Thus, we aimed to investigate the impact of NETs in ERU patients. Therefore, we quantified several NET-markers (cell-free DNA, nucleosomes, citrullinated histone H3, histone-myeloperoxidase complexes, interleukin-17, equine cathelicidin 1 and DNase I activity) and NET-autoantibodies in sera and vitreous body fluids (VBF) of ERU-diseased horses and correlated the data with the disease status (signalment, ERU scores and Leptospira infection status). NET markers were detected to varying degrees in VBF of diseased horses, and partially correlated to disease severity and the presence of Leptospira spp. Cell-free DNA and nucleosomes as NET markers correlate with ERU severity in total and VBF scores, despite the presence of active DNases. Additionally, a significant correlation between fundus affection in the eye and NET autoantibodies was detectable. Therefore, we further investigated the influence of VBF samples from equine patients and isolated NETs on the blood-retina barrier in a cell culture model. VBF of diseased horses significantly induced cytotoxicity in retinal pigment epithelial cells. Moreover, partially digested NETs also resulted in cytotoxic effects. In the presence of lipopolysaccharide (LPS), the main component of the leptospiral surface, both undigested and completely digested NETs were cytotoxic. Correlations between the ERU-scores and Leptospira were also calculated. Detection of leptospiral DNA, and antibody titers of the serovar Grippotyphosa correlated with disease severity. In addition, a correlation between Leptospira and several NET markers was observed in VBF. Altogether, our findings suggest a positive correlation between NET markers with disease severity and involvement of Leptospira in the VBF of ERU-diseased horses, as well as a cytotoxic effect of NETs in eyes., Competing Interests: Author KS-M is employed by IVD Innovative Veterinary Diagnostics (IVD GmbH), Seelze, Germany. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Fingerhut, Yücel, Strutzberg-Minder, von Köckritz-Blickwede, Ohnesorge and de Buhr.)

Ophthalmic problems account for up to 20% of emergencies in equine practice. Presenting problems may involve the periocular region, adnexa, or globe. Practitioners must have the experience to restrain horses with painful ocular conditions and the knowledge of how to perform a thorough eye examination. A range of clinical skills is required for the necessary diagnostic tests such as corneal cytology, ultrasound and tonometry, and common standing surgical procedures such as eyelid repair or instillation of a subpalpebral lavage system. Therapy, which may involve frequent administration of multiple medications, must be targeted to the specific diagnosis., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Objective: Equine recurrent uveitis (ERU), a chronic, immune-mediated intraocular inflammatory disease, is a common cause of blindness in horses. The severity and recurrent nature of ERU makes it difficult to treat with current therapeutics leading to a poor visual prognosis. The suprachoroidal space (SCS), a potential space between the choroid and sclera surrounding the ocular posterior segment, offers a promising alternative site for drug application to the eye. Corticosteroid administration within this space is hypothesized to be safe and effective at controlling intraocular inflammation, especially in horses with poorly responsive ERU., Animal Studied: Horses with active, poorly responsive ERU., Procedure(s): A retrospective study was performed with 29 horses (36 total eyes) that received SCS injection of triamcinolone acetonide (TA) with ERU not well controlled with standard uveitis treatment. A standardized ocular inflammation score (OIS) was used to assess inflammation at the time of injection and at follow-ups., Results: Standardized OIS revealed a significant decrease in ocular inflammation over time after SCS TA administration (p < .004). Adverse effects after injections occurred in <20% of the horses at follow-up, but some of these effects were attributed to chronic inflammation prior to effective treatment, long-term topical corticosteroid use, or complications from hospitalization rather than the SCS injections. Most horses (86.7%) in this study remained visual greater than 3 months after SCS injection., Conclusions: Based on these results, SCS TA injections appear to be a safe and possible effective treatment modality for managing poorly responsive ERU; further clinical study is warranted., (© 2021 American College of Veterinary Ophthalmologists.)

Equine recurrent uveitis (ERU) is a spontaneous, remitting-relapsing autoimmune disease driven by the adaptive immune system. Although T cells are described as the main effector cells in pathogenesis, granulocytes have also emerged as possible disease mediators. To explore the role of these innate immune cells, we investigated the whole cell proteome of granulocytes from equine recurrent uveitis cases and healthy controls. Among the 2362 proteins identified by mass spectrometry, we found 96 proteins with significantly changed abundance between groups (p < 0.05, fold change >1.2), representing 4.1% of total granulocyte proteome. Within these differential identifications, calgranulin B, a protein associated with pathogenesis in other autoimmune diseases, showed highest abundance in equine recurrent uveitis (18 fold). For a better interpretation of the results from our hypothesis-generating approach, we added a threshold for biological significance (ratio ERU/controls >2: 36 proteins) to the proteins with increased abundance in equine recurrent uveitis and analyzed their allocation to the subsets within the Immune System superpathway. The 36 differentially abundant proteins predominantly associated to RAF/MAP kinase cascade, MHC-I-mediated antigen presentation and neutrophil degranulation, suggesting a latently activated phenotype of these innate immune cells in disease. Raw data are available via ProteomeXchange with identifier PXD013648. SIGNIFICANCE: Our study provides new insights into the protein repertoire of primary equine granulocytes and identifies protein abundance changes associated to equine recurrent uveitis (ERU), an organ specific, spontaneously occurring autoimmune disease. We show that granulocyte proteins with increased abundance in ERU strongly associate to RAF/MAP kinase signaling, MHC-I antigen presentation and neutrophil degranulation, pointing to a more activated state of these cells in ERU cases. Since cells were obtained in quiescent stage of disease, latent activation of granulocytes underlines the role of these innate immune cells in ERU. These findings are highly relevant for veterinary medicine, further establishing the importance of granulocytes in this T cell-driven autoimmune disease. Moreover, they have translational quality for autoimmune uveitis in man, due to strong similarity in disease occurrence, progression and pathogenesis., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Uveitis is reported to be a leading cause of blindness in horses. Little work has been undertaken to date to investigate whether donkeys are affected by a similar disease prevalence., Objectives: To investigate the disease prevalence of uveitis in a population of donkeys in the UK., Study Design: Descriptive observational study., Methods: An ophthalmic examination was performed on each donkey, in a darkened stable. Each donkey underwent slit lamp biomicroscopy, and direct and/or indirect ophthalmoscopy. Fluorescein staining, STT1 and IOPs were measured when deemed clinically necessary. Pharmacological pupillary dilation was achieved using 1% tropicamide., Results: A total of 207 donkeys were examined: 139 males (67.1%) and 68 females (32.9%). Age range was 2-37 years (median: 17 years, interquartile range: 9-25 years). Three donkeys (1.5%) were blind in one eye, and one was monocular at the time of examination. Signs consistent with either previous or current uveitis were identified in eight eyes of six animals (2.9%). Clinical signs included are as follows: miosis (n = 1), corpora nigra atrophy (n = 6), anterior lens capsule pigment (n = 2), cataract (n = 8), posterior synechiae (n = 3), lens subluxation (n = 1), vitreal changes (n = 2), peripapillary scarring (n = 3) and phthisis bulbi (n = 1). Significant ocular pathology precluded fundic examination in three eyes. Three out of eight eyes (37.5%) were blinded by the pathology. The risk of uveitis was found to increase with age (OR 1.1, CI: 1.01-1.25) on univariable analysis (P = .046.)., Main Limitations: Fluorescein staining, tonometry and Schirmer tear test measurements were not performed on all donkeys., Conclusions: The disease prevalence reported here is comparable to that reported for horses in the UK. Larger prospective studies are necessary to determine whether there is a similar trend in disease prevalence in the donkey population worldwide. In addition, further investigations should aim to establish whether uveitis in donkeys is similar to the ERU syndrome seen in horses., (© 2020 EVJ Ltd.)

Objective: To evaluate leptospiral antibody prevalence in 65 horses with ERU and compare outcome in 36 surgically treated eyes (2010-2015)., Procedures: Retrospective data analysis of horses with ERU (n = 65). C-value calculation with microagglutination assay titer (MAT) results for Leptospira spp. Evaluation of follow-up data after pars plana vitrectomy (PPV, n = 21 eyes) and suprachoroidal cyclosporine device implantation (SCDI, n = 15 eyes). Differences between groups were statistically analyzed using Fishers exact test, significance set at P < .05., Results: Positive leptospiral titers were found in 28/65 blood, 31/65 aqueous humor (AH), and 19/20 vitreal (post-PPV) samples. The most common intraocular serovars were Leptospira interrogans grippotyphosa, pomona, and bratislava. Intraocular antibody production was suspected in samples of 22 horses (c-values > 1). Mean follow-up of surgical cases was 3.8 years (PPV) and 3.4 years (SCDI). PPV was performed in 21 eyes with positive, SCDI in 15 eyes with negative leptospiral test results. Uveitis recurred less often after PPV (2/21) compared to SCDI (6/15, P = .04). Retinal detachment occurred after PPV only (5/21, SCDI 0/15, P = .06), whereas only SCDI-treated eyes were enucleated (PPV 0/21, SCDI 3/15, P = .06). Blindness or visual impairment was equally likely to occur in both treatment groups after surgery (PPV 7/21, SCDI 7/15, P = .5)., Conclusions: Leptospiral antibody prevalence is high in horses with ERU in Switzerland. Recurrence of uveitis is uncommon following PPV in the present study; an increased risk of retinal detachment exists. Enucleation is more often warranted in horses after SCDI in this study due to a higher uveitis recurrence., (© 2020 American College of Veterinary Ophthalmologists.)

Objective: To characterize clinical manifestations, measure frequency, and evaluate risk factors for equine recurrent uveitis (ERU) in Appaloosa horses in western Canada., Animals: 145 Appaloosa horses., Procedures: Ophthalmic examinations were completed and eyes were classified as having no or mild clinical signs, or moderate, or severe damage from ERU. Clinical signs, age, sex, base coat color, and pattern were recorded. Whole blood and/or mane hair follicles were collected for DNA extraction, and all horses were tested for the leopard complex (LP) spotting pattern allele. Pedigree analysis was completed on affected and unaffected horses, and coefficients of coancestry (CC) and inbreeding (COI) were determined., Results: Equine recurrent uveitis was confirmed in 20 (14%) horses. The mean age of affected horses was 12.3 years (±5.3; range 3-25). Age was a significant risk factor for ERU diagnosis (OR year  = 1.15) and classification (OR year  = 1.19). The fewspot coat pattern was significantly associated with increased risk for ERU compared to horses that were minimally patterned or true solids. The LP/LP genotype was at a significantly greater risk for ERU compared to lp/lp (OR = 19.4) and LP/lp (OR = 6.37). Classification of ERU was greater in the LP/LP genotype compared to LP/lp. Affected horses had an average CC of 0.066, and there was a significant difference in the distribution of CC for affected horses versus the control group (P = .021). One affected horse was the sire or grandsire of nine other affected., Conclusions: Age, coat pattern, and genetics are major risk factors for the diagnosis and classification of ERU in the Appaloosa., (© 2020 American College of Veterinary Ophthalmologists.)

Equine recurrent uveitis (ERU) is characterized by intraocular inflammation that often leads to blindness in horses. Appaloosas are more likely than any other breed to develop insidious ERU, distinguished by low-grade chronic intraocular inflammation, suggesting a genetic predisposition. Appaloosas are known for their white coat spotting patterns caused by the leopard complex spotting allele (LP) and the modifier PATN1. A marker linked to LP on ECA1 and markers near MHC on ECA20 were previously associated with increased ERU risk. This study aims to further investigate these loci and identify additional genetic risk factors. A GWAS was performed using the Illumina Equine SNP70 BeadChip in 91 horses. Additive mixed model approaches were used to correct for relatedness. Although they do not reach a strict Bonferroni genome-wide significance threshold, two SNPs on ECA1 and one SNP each on ECA12 and ECA29 were among the highest ranking SNPs and thus warranted further analysis (P = 1.20 × 10 -5 , P = 5.91 × 10 -6 , P = 4.91 × 10 -5 , P = 6.46 × 10 -5 ). In a second cohort (n = 98), only an association with the LP allele on ECA1 was replicated (P = 5.33 × 10 -5 ). Modeling disease risk with LP, age and additional depigmentation factors (PATN1 genotype and extent of roaning) supports an additive role for LP and suggests an additive role for PATN1. Genotyping for LP and PATN1 may help predict ERU risk (AUC = 0.83). The functional role of LP and PATN1 in ERU development requires further investigation. Testing samples across breeds with leopard complex spotting patterns and a denser set of markers is warranted to further refine the genetic components of ERU., (© 2019 Stichting International Foundation for Animal Genetics.)

Background: Uveitis appears to be less prevalent in the UK compared with other parts of the world and studies characterising the disease in the UK are lacking. The objectives of this retrospective study were to describe acute and recurrent cases presenting for management of uveitis in a referral hospital on the North West of England and compare the signalment of horses presenting with uveitis with the equine hospital population during the same period., Methods: Medical records of horses presented to the referral Equine Hospital, University of Liverpool with signs of uveitis between 2008 and 2018 were reviewed and clinical details extracted., Results: Seventy horses presented with uveitis; 33 were classified as acute and 37 as recurrent cases. Sixteen of the horses were affected bilaterally. More bilateral cases were classified as recurrent than acute (P=0.04). No differences in age or sex were noted between acute and recurrent cases, or between cases and the general hospital population. Warmbloods and Appaloosas were over-represented when compared with the general hospital population (P<0.001). Twenty-one horses (30 per cent, 95 per cent CI 20.5 to 41.4) underwent surgery for the control of the uveitis. Fourteen of the 70 horses (20.0 per cent, 95 per cent CI 12.3 to 30.8) underwent enucleation., Conclusion: While relatively uncommon in the UK, uveitis can affect horses from a relatively young age. The disease appears to have a relatively higher frequency than expected in Warmbloods and Appaloosas. It is more likely that a recurrent case will have both eyes affected., Competing Interests: Competing interests: None declared., (© British Veterinary Association 2019. No commercial re-use. See rights and permissions. Published by BMJ.)

Equine recurrent uveitis (ERU) is an immune-mediated disease causing repeated or persistent inflammatory episodes which can lead to blindness. Currently, there is no cure for horses with this disease. Mesenchymal stem cells (MSCs) are effective at reducing immune cell activation in vitro in many species, making them a potential therapeutic option for ERU. The objectives of this study were to define the lymphocyte phenotype of horses with ERU and to determine how MSCs alter T-cell phenotype in vitro. Whole blood was taken from 7 horses with ERU and 10 healthy horses and peripheral blood mononuclear cells were isolated. The markers CD21, CD3, CD4, and CD8 were used to identify lymphocyte subsets while CD25, CD62L, Foxp3, IFNγ, and IL10 were used to identify T-cell phenotype. Adipose-derived MSCs were expanded, irradiated (to control proliferation), and incubated with CD4 + T-cells from healthy horses, after which lymphocytes were collected and analyzed via flow cytometry. The percentages of T-cells and B-cells in horses with ERU were similar to normal horses. However, CD4 + T-cells from horses with ERU expressed higher amounts of IFNγ indicating a pro-inflammatory Th1 phenotype. When co-incubated with MSCs, activated CD4 + T-cells reduced expression of CD25, CD62L, Foxp3, and IFNγ. MSCs had a lesser ability to decrease activation when cell-cell contact or prostaglandin signaling was blocked. MSCs continue to show promise as a treatment for ERU as they decreased the CD4 + T-cell activation phenotype through a combination of cell-cell contact and prostaglandin signaling., (© 2019 The Authors. Veterinary Ophthalmology published by Wiley Periodicals, Inc. on behalf of American College of Veterinary Ophthalmologists.)

We report a retrospective evaluation of intravitreal gentamicin (6 mg in 0.3 mL solution) injection in 71 horses with equine recurrent uveitis between 2010 and 2016. Aqueous flare and hyalitis were scored at the baseline in all horses, then at 15, 30, 60, 90, 180, 365, and 730 in 37, 46, 30, 25, 22, 18, and 11 horses, respectively. Forty-two animals had a global score of inflammation (sum of scores for aqueous flare and hyalitis) of zero at the last examination (number of re-examinations per horse ranged from 1 to 5). Among the 29 remaining horses, four were not reassessed after the intervention. The individual inflammation scores of the remaining 25 horses improved in 21 cases, whereas it slightly deteriorated in four cases (number of re-examinations ranged from 0 to 6). Ocular hypotension, lens opacities, and retinal folds were observed, respectively, in 92%, 78%, and 28% of the 71 eyes at the baseline, and in 80%, 90%, and 36% of the 11 eyes re-evaluated at 2 years. A telephone inquiry about all horses among owners and referring veterinarians conducted in 2016 at least 6 months after the more recent injections indicated that 70 horses did not develop a new observable episode of inflammation and did not need ocular medication. Although the results of this study should be discussed with reservation, it indicated that low-dose intravitreal injection of gentamicin could be a useful treatment to maintain ocular quiescence in equine recurrent uveitis., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: Equine recurrent uveitis (ERU) has been associated with Leptospira spp. infection. No information exists concerning the prevalence of Leptospira-associated ERU in Belgium and about the sensitivity of detection of Leptospira in different ocular media., Objectives: To establish the prevalence of intraocular Leptospira spp. in ERU-affected and healthy eyes of horses examined at the Equine Clinic of the University of Liège by real-time PCR and to compare the results of the aqueous and vitreous humour of the same eye., Study Design: Cross-sectional., Methods: Sixty-six eyes from 59 client-owned horses with a diagnosis of equine recurrent uveitis (ERU-group) were studied from May 2015 to December 2017. Fifty healthy eyes from 28 euthanised horses for unrelated reasons examined during the same period were included in the control group. Intraocular fluids (aqueous and/or vitreous humours) from ERU-affected eyes were sampled and analysed by real-time PCR for Leptospira spp. Aqueous and vitreous humours from the control group were processed in the same way., Results: Both groups were comparable regarding age, sex, eye sampled (OS/OD), humours sampled (aqueous/vitreous humour) but not regarding breeds, with an over-representation of Warmbloods and Appaloosas in the ERU-group. The prevalence of Leptospira spp. was 30.3% (20/66 eyes) in the ERU-group. Leptospira spp. DNA was identified in 11 aqueous and 17 vitreous humours with eight horses testing positive in both humours, nine horses testing positive for vitreous humour alone and third horses for aqueous humour alone. The phi-correlation between aqueous and vitreous humour Leptospira-PCR results is 0.47 suggesting a low association. All the control eyes were negative., Main Limitations: The diagnostic method selected for this study (lipL32 qPCR) did not allow identification of the serovars., Conclusions: Leptospirosis is a potential cause of ERU in Belgium. Testing both intraocular media is advised whenever possible. The Summary is available in Spanish - see Supporting Information., (© 2018 EVJ Ltd.)

Objective: Aim of the study was the retrospective evaluation regarding vision and recurrence in equine recurrent uveitis (ERU)-affected horses treated by intravitreal low-dose gentamicin injection in Tierklinik Alt Sammit during the years 2006-2013., Material and Methods: The study included 50 equine patients (61 ERU-affected eyes) of different age and breed with different stages of ERU. The solution for the injection contained 4.0 mg sterile gentamicin per eye. The injection was performed according to the procedure described by Pinard et al. in 2005 under general anaesthesia in lateral recumbency of the patient and within a non-inflammatory period. The patients were monitored over a postoperative period of 2-96 months (mean 25.8 months, median 15 months)., Results: A total of 56 out of 61 treated eyes displayed no recurrence of ERU during the follow-up period. Recurrence of a single uveitic attack occurred in five eyes (8.2 %) within a period of 2 weeks to 28 months after surgery. Long-term stable vision was achieved in 43 out of 61 eyes (70.5 %). Postoperative blindness developed in 18 eyes, the majority hereof had already preoperatively displayed obvious pathologic ophthalmic findings with partly highly impaired vision. The main reason for a vision loss was the progression of a preoperatively existing cataract formation. Perioperative complications were limited to vitreous reflux into the subconjunctival space (two cases), subconjunctival haemorrhage in the area of the injection site (two cases), vitreous haemorrhage (one case) and two cases of immediate postoperative pain reaction associated with the intravitreal injection., Conclusion and Clinical Relevance: The results of the therapy regarding the reduction of the recurrence rate and preservation of vision are comparable to those of vitrectomy. The complication rate is extremely low when using both the correct injection technique and amount of gentamicin solution. Therefore, the intravitreal injection of low-dose gentamicin is an inexpensive and uncomplicated alternative to vitrectomy., Competing Interests: Die Autoren bestätigen, dass zu den Inhalten dieser Arbeit kein Interessenkonflikt besteht., (© Georg Thieme Verlag KG Stuttgart · New York.)

Background: Despite appropriate medical therapy, many horses with equine recurrent uveitis continue to suffer from recurrent bouts of inflammation. Surgical intervention via the pars plana vitrectomy or suprachoroidal cyclosporine implant placement may control and/or prevent recurrences, however, these procedures may be contraindicated, unavailable, or declined by an owner. Thus, an effective adjunctive treatment option may help to improve the clinical outcomes in those situations. There are several anecdotal reports on the use of intravitreal gentamicin injections, but to date, no data evaluating the complication rate and/or treatment effect following this treatment have been published. Thus, the aim of this prospective study was to describe the intravitreal gentamicin injection technique, describe the associated peri-injection (within 24 h) and post-injection (30 to 780 days) complications, and to report the effects of the injection on the clinical signs of uveitis. Additionally, evaluation of the systemic and ocular Leptospira-status, and its effect on the treatment outcome was performed. A total of 86 horses of various ages, breeds, and gender presenting with recurrent or persistent uveitis were treated via intravitreal injection of 4 mg of undiluted gentamicin (0.04 ml, Genta 100, 100 mg/ml in 35 horses) or preservative-free gentamicin (0.05 ml, 80 mg/ml in 52 horses) under sedation and local anesthesia. All 86 horses were observed for immediate peri-injection and post-injection complications. Response to therapy was evaluated in 59 of the 86 horses (follow-up: 30 to 780 days)., Results: Peri-injection complications consisted of subconjunctival (26/86; 30.2%) or intracameral hemorrhage (4/86; 4.7%); both of which completely resolved within 5 days. Post-injection complications consisted of cataract formation/maturation (5/59 horses, 8.5%) and diffuse retinal degeneration (3/59 eyes 5.1%). The majority of horses 52/59 (88.1%) with a minimum follow-up period of 30 days were controlled (absence of recurrent or persistent inflammation) at their last recheck examination. Recurrent inflammation was documented in 5/59 (8.5%) horses and persistent inflammation was diagnosed in 2/59 (3.4%) horses., Conclusions: Intravitreal injection of low-dose gentamicin shows promise at controlling different types and stages of uveitis. The ability of intravitreal injections of low-dose gentamicin (4 mg) to control persistent and recurrent inflammation warrants further investigation.

Rhodococcus equi is one of the most important causes of disease in foals. Infection is typically characterized by pyogranulomatous pneumonia although extrapulmonary infections occur occasionally. Uveitis and polysynovitis have been reported in foals naturally infected with R. equi and are thought to be the result of an immune-mediated process. However, the pathogenesis of these conditions is poorly understood. The objectives of this study were to document the occurrence of uveitis and polysynovitis after experimental infection with R. equi and to determine if these disorders are the direct result of infection at these sites. Foals between 3 and 4 weeks of age were infected intratracheally with virulent R. equi using inocula of 1×108 CFU (high inoculum; n = 16) or 1×107 CFU (low inoculum; n = 12). Foals were monitored twice daily and necropsy was performed 14 days post-infection. Aqueous humor and synovial fluid were collected aseptically and the percentage of affected lung was calculated. The mean (± SD) percentage of affected lung was significantly higher with the high inoculum (31.8 ± 14.6%) than with the low inoculum (14.4 ± 11.4%). Fourteen of 25 foals developed uveitis and 20 of 28 foals developed polysynovitis. R. equi was cultured from the aqueous humor of 11 foals and from the synovial fluid of 14 foals. The risk of development of polysynovitis and protein concentration in the aqueous humor were significantly higher in foals that received the high inoculum. These results indicate that polysynovitis and uveitis are septic complications associated with the severity of lung disease.

Equine recurrent uveitis (ERU) is characterized by recurrent bouts of inflammation interrupted by periods of quiescence that vary in duration. There is little consensus on the clinical manifestations, the underlying causes, or the management. The 3 commonly recognized syndromes of ERU (classic, insidious, and posterior) do not accurately separate the clinical manifestations of disease into distinct categories. An accurate diagnosis and early intervention are essential to minimizing the effects of disease and preserving vision. There are multiple medical and surgical options for controlling ERU as long as the disease is recognized early and targeted treatment is initiated immediately., (Copyright © 2017 Elsevier Inc. All rights reserved.)

A recently purchased 16-year-old Thoroughbred mare with a history of recurrent colic and low body weight, also had lameness in 3 limbs, pain at the sacro-iliac joint, and increased corneal opacity of the right eye. Response to supportive therapy was poor and euthanasia was elected. A postmortem examination and histopathology confirmed cyathostomiasis within the large colon and cecum and chronic anterior uveitis of the right eye., Competing Interests: Use of this article is limited to a single copy for personal study. Anyone interested in obtaining reprints should contact the CVMA office (hbroughton@cvma-acmv.org) for additional copies or permission to use this material elsewhere.

Background: Equine recurrent uveitis (ERU) is a common cause of ocular pain and blindness in horses. Leptospira spp. have been commonly implicated in the pathophysiology of ERU in mainland Europe and the USA. No recent studies have been carried out in the UK, but Leptospira is reported not to be a major factor in the aetiology of ERU in the UK., Objectives: To establish the prevalence of Leptospira-associated ERU in the UK and to identify the serovars involved in these cases; to compare serum vs. aqueous humour antibody levels in cases and controls in order to confirm the diagnosis of Leptospira-associated ERU, and to assess the usefulness of serology alone as a confirmatory test for Leptospira-associated ERU in the UK., Study Design: Case-control study., Methods: Eyes enucleated for clinical reasons in ERU-affected horses were collected. Blood and aqueous humour were obtained to determine antibody levels against a variety of Leptospira serovars and C-values (aqueous humour value/serum value) were calculated. In addition, eyes, blood and aqueous humour were obtained from control cases for comparison. Histopathology was performed in all eyes to confirm uveitis in each case. Differences in seroprevalences between ERU and control cases and between Leptospira- and non-Leptospira-associated ERU cases were calculated., Results: A total of 30 ERU and 43 control eyes were analysed. Of the ERU eyes, only two had a C-value of >4 (prevalence of Leptospira-associated uveitis: 6.7%). Serovars hardjo and javanica were detected. There was no difference in seroprevalence between horses with uveitis and control cases (65.5% and 41.9%, respectively; P = 0.11) or between Leptospira- and non-Leptospira-associated uveitis cases (100% and 63.0%, respectively; P = 0.52)., Main Limitations: The study was limited by low case numbers. Eyes were presented at different stages of disease. The only test used to detect Leptospira was the microscopic agglutination test., Conclusions: Leptospira-associated ERU is uncommon in the UK. Serology alone may not help to definitively diagnose Leptospira-associated uveitis in this country., (© 2017 The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.)

Equine recurrent uveitis is the only spontaneous model for recurrent autoimmune uveitis in humans, where T cells target retinal proteins. Differences between normal and autoaggressive lymphocytes were identified in this study by analyzing peripheral blood derived lymphocytes (PBL) proteomes from the same case with interphotoreceptor retinoid binding protein induced uveitis sampled before (Day 0), during (Day 15), and after uveitic attack (Day 23). Relative protein abundances of PBL were investigated in a quantitative, label-free differential proteome analysis in cells that were kept frozen for 14 years since the initial experiment. Quantitative data could be acquired for 2632 proteins at all three time points. Profound changes (≥2-fold change) in PBL protein abundance were observed when comparing Day 0 with 15, representing acute inflammation (1070 regulated proteins) and Day 0 with 23 (cessation; 1571 regulated). Significant differences applied to proteins with functions in integrin signaling during active uveitis, involving "Erk and pi-3 kinase are necessary for collagen binding in corneal epithelia," "integrins in angiogenesis," and "integrin-linked kinase signaling" pathways. In contrast, at cessation of uveitic attack, significantly changed proteins belonged to pathways of "nongenotropic androgen signaling," "classical complement pathway," and "Amb2 integrin signaling." Several members of respective pathways were earlier shown to be changed in naturally occurring uveitis, underscoring the significance of these findings here and proofing the value of the induced model in mimicking spontaneous autoimmune uveitis. All MS data have been deposited to the ProteomeXchange consortium via the PRIDE partner repository (dataset identifier PXD005580)., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The objectives of this study were to determine the demographics of horses with equine recurrent uveitis (ERU) presenting to the Western College of Veterinary Medicine and to describe and compare the prognosis of ERU in the Appaloosa with that in other breeds. Horses diagnosed with ERU by a veterinary ophthalmologist between 2002 and 2015 were included. Eye lesions were classified as mild, moderate, or severe based on clinical manifestations. Breed, age, severity, blindness, and final outcome were evaluated. Thirty-two horses fit the inclusion criteria; 62.5% were Appaloosas. Mean age at presentation was 12.13 ± 4.6 years. Equine recurrent uveitis was bilateral in 93.6% of horses and was severe in 59.4% of eyes at presentation. Bilateral blindness was present in 59.4% of horses at last follow-up. Of 27 horses available for follow-up, 63% were euthanized due to ERU. No significant differences in age, severity, blindness, or rate of euthanasia were noted between Appaloosas and other breeds. The Appaloosa is at increased risk for ERU, which is a devastating ocular disease.

The membrane protein expression repertoire of cells changes in course of activation. In equine recurrent uveitis (ERU), a spontaneous autoimmune disease in horses with relapsing and ultimately blinding inner eye inflammation, CD4+ T lymphocytes are the crucial pathogenic cells activated in the periphery directly prior to an inflammatory episode. In order to find relevant changes in the membrane proteome associated to disease, we sorted CD4+ lymphocytes and compared protein abundance from the generated proteome datasets of both healthy horses and ERU cases. We detected formin like 1, a key player in actin dependent cellular processes such as phagocytosis, cell adhesion and cell migration, with significantly higher abundance in the CD4+ cell membrane proteome of horses with ERU. In transmigration experiments, we demonstrated higher migration rate of cells originating from diseased animals connecting formin like 1 to the migratory ability of cells. These findings are the first description of formin like 1 in association to processes involved in migration of inflammatory CD4+ T cells across the blood-retinal barrier in a spontaneous ocular autoimmune disease and suggest formin like 1 to play a role in the molecular mechanisms of ERU disease pathogenesis. Data are available via ProteomeXchange with identifier PXD005384., Biological Significance: This comparative proteomic study of membrane proteins of CD4+ T cells identified a novel protein, formin like 1, with expression on the plasma cell membrane of equine CD4+ T cells and a significant change of abundance on CD4+ T cells of horses with a spontaneous autoimmune disease. Functional studies in a cell culture model for transmigration at the blood-retinal barrier (BRB) unraveled a strong impact of formin like 1 on migratory processes of CD4+ T cells across the BRB, a key event in uveitis pathogenesis. These findings provide novel knowledge about changes in the CD4+ immune cell membrane proteome in a spontaneously and naturally occurring autoimmune disease in horses with high relevance for veterinary medicine. Additionally, this model has proven translational quality for human medicine and provides novel proteomic information on autoimmune uveitis in man., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: The pathogenic mechanism of equine recurrent uveitis (ERU) is still poorly defined and many variations between experimental animal models and spontaneous disease exist., Objectives: The aim of our study was to investigate if Th17 cell-mediated response plays role in the pathogenesis of the used experimental model in horses and to reveal its pathological findings., Methods: Experimental uveitis was induced in 6 healthy horses. The concentrations of retinal autoantigen CRALBP and IL-17 were measured using ELISA in aqueous humor and vitreous body of the 12 inflamed eyes as well as in 12 control non-inflamed eyes taken from 6 horses in slaughter house. After centrifugation of the two eye media, smears were prepared and cytological investigation was performed. Tissue specimens were taken from all eye globes and were submitted to histopathological investigation., Results: CRALBP and IL-17 concentrations were significantly elevated in eye media of horses with experimental uveitis in comparison with controls. Cytological and histopathological findings corresponded to the changes characteristic of chronic immune-mediated inflammation with mononuclear cell infiltration of uvea, choroid, retina, and eye media as well as severe retinal destruction., Conclusions: Our study demonstrated the involvement of the retinal autoantigen CRALBP as well as IL-17 in the pathogenesis of experimental uveitis in horses. These findings suggests that this experimental uveitis in horses may serve as a suitable animal model for investigation of IL-17- mediated immune response during spontaneous autoimmune uveitis in horses as well as in humans.

Objective: To determine whether horses with clinically diagnosed Equine Recurrent Uveitis (ERU) and those with Leptospirosis infection have a specific cytokine profile in their aqueous humor (AH) and serum that differs from horses with uveitis secondary to other ocular inflammatory processes and from horses with normal eyes., Animals Studied: Twenty-five client-owned horses with uveitis that were presented to the North Carolina State University Ophthalmology Service, and four University-owned horses without history or clinical signs of ocular disease., Procedure: Samples of AH and serum were obtained from horses with ERU (n=13), acute or non-recurrent uveitis (UV; n=7), uveitis secondary to infectious keratitis (IK; n=5), and normal eyes (N; n=4). Cytokine levels in AH and serum were quantified using a multiplex bead immunoassay. Leptospiral antibody titers in serum and AH and PCR for Leptospiral DNA in AH were performed., Results: In the AH of horses with ERU, increased levels of IL-1a, IL-4, IL-6, IL-8, IL-12p70, FGF-2, G-CSF, and RANTES were measured compared to UV, IK and N eyes, but the differences were not significant. However, IL-10 was significantly higher in ERU eyes compared to IK and N (P=0.029; 0.013), and IP-10 in ERU eyes was significantly higher than in UV and N (P=0.004). Furthermore, MCP-1 was significantly higher in ERU than N (P=0.04). In the serum, increased levels of IL-1a, IL-4, IL-6, IL-8, IL-12p70, fractalkine, and G-CSF were measured in horses with ERU, but the levels were not significantly higher than those observed in UV, IK, or N horses. However, serum IP-10 levels in horses with ERU were significantly higher than in UV and N horses (P=0.005) and MCP-1 levels were significantly higher in ERU than N (P=0.03). Horses with marked ocular inflammation had significantly higher serum levels of G-CSF, IL-1a, fractalkine, IL-13, IL-4, IL-17a, IL-12p70, IFN-γ, and MCP-1. Elevated IL-10 in AH was significantly associated with disease chronicity, both overall and in ERU eyes (P=0.049), and in horses with positive ocular leptospiral titers or leptospiral PCR, significant elevations of IL-10 (P=0.0018; 0.0032) and IP-10 (P=0.0342; 0.043) were detected in the AH compared to leptospiral negative eyes., Conclusions: The anti-inflammatory cytokine IL-10 and the pro-inflammatory cytokine IP-10 appear to play an important role in ERU. Further studies are needed to further clarify and characterize cytokine profiles of specific ocular inflammatory diseases, but multiplex bead immunoassay technology shows promise as a diagnostically valuable tool., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Reasons for Performing Study: Equine recurrent uveitis (ERU) is a leading cause of vision loss in horses., Objectives: To assess the prognosis and impact of ERU on affected horses and their owners by evaluating the signalment, treatment and outcome (including the loss of use, vision assessment and economic loss)., Study Design: Retrospective impact study., Methods: Medical records of horses presenting to the North Carolina State University Veterinary Health Complex (NCSU-VHC) with ERU between 1999 and 2014 were reviewed. Signalment, clinical signs, ophthalmic examination findings, treatments and outcomes were evaluated. Owner questionnaires were completed regarding vision, job/role, monetary value, diagnostic and treatment costs, concurrent illness and outcomes., Results: Records of 224 horses (338 eyes) with ERU were reviewed. There was an overrepresentation of Appaloosas (54; 24.1%), Hanoverians (11; 4.9%) and other Warmbloods (13; 5.8%). Ninety-six eyes (28.4%) were diagnosed as blind and 38 eyes (11.2%) with glaucoma on initial evaluation. Leptospirosis titres of serum and/or aqueous humour were obtained in 88 horses and were positive in 40 horses (45.5%), with L. pomona being the most frequently isolated serovar. Globe loss at the NCSU-VHC occurred in 41 ERU eyes (12.1% of total). Owner questionnaires were evaluated in 194 horses (86.6%) and 91 horses (46.9%) were reported blind in the affected eye(s). Fifty-seven horses (29.4%) did not return to their previous role, while 61 (31.4%) performed at a reduced level. Equine recurrent uveitis decreased the monetary value of 164 horses. Twenty-nine horses (14.9%) were euthanised and 37 (19.1%) underwent change in ownership as a direct result of ERU., Conclusions: The impact of ERU is attributed to the high frequency of blindness, globe loss and loss of function. Euthanasia and change of ownership are common sequelae to the progressive nature of ERU. These factors, along with financial costs of the disease, have a significant impact on affected horses and their owners., (© 2015 EVJ Ltd.)

In the ongoing discussion regarding the aetiopathogenesis of equine recurrent uveitis (ERU) it was the aim of the present study to elucidate the relationship of leptospira infection and ERU. In a population of 225 horses leptospira were examined in vitreous humor by culture and leptospira antibody were detected in vitreous humor and serum samples. Preoperative serum samples were collected from 221/225 ERU patients of different age, gender and breed. Undiluted vitreous humor was aseptically taken from 198/225 patients that underwent pars plana vitrectomy at the beginning of surgery and from 27/225 patients' eyeball after enucleation: Serum and vitreous humor were tested for specific leptospiral antibodies by microscopic agglutination test (MAT). Furthermore, vitreous humor was examined by culture. 20 patients which were euthanized due to a live-threatening disease other than ERU served as a control group. A total of 127/221 (57.5%) horses had serum antibodies (≥ 1:100). Most frequently antibodies against L. interrogans serovar Grippotyphosa were detected (79/127), followed by L. interrogans serovar lcterohaemorrhagiae (34/127) and L. interrogans serovar Bratislava (29/127). Only 79/225 horses (35.1%) had leptospiral antibodies in vitreous humor, in which L. interrogans serovar Grippotyphosa (67/79) was identified most frequently followed by L. interrogans serovar Pomona (18/79) and L. interrogans serovar lcterohaemorrhagiae (8/79) which was identified as single or multiple reaction. Isolation of leptospira from vitreous humor was positive in 34/212 horses (16%). 10/20 control horses had a positive antibody titer against leptospira in serum and 2/20 horses in vitreous humor, whereas there was no leptospira detected in culture. The result of 84% negative cultures from vitreous humor of 212 ERU patients is decisive for the diagnosis and therapy of ERU.

Equine recurrent uveitis (ERU) has various etiologies, with Leptospira infection and genetic predisposition being the leading risk factors. Regardless of etiology, expression of ocular proteins associated with maintenance of the blood-ocular barrier is impaired in ERU. The recurring-remitting cycle of ERU repeatedly disrupts the blood-ocular barrier, allowing the previously immune-privileged ocular environment to become the site of a progressive local autoimmune pathology that ultimately results in tissue destruction and vision loss. The immune-mediated process involves humoral and cellular mechanisms. Intraocular antibodies either produced in the eye or that leak through the blood-ocular barrier, are often present at higher levels than in serum and react with antigens in ocular tissue of horses with ERU. Ocular infiltration of auto-aggressive lymphocytes occurs with each uveitis episode and is the most crucial contributor to inflammation and eye damage. Recurring uveitis episodes may be initiated when epitopes of an ocular antigen become visible to the immune system (intramolecular spreading) or another autoantigen (intermolecular spreading), resulting in a new inflammatory reaction., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Equine recurrent uveitis (ERU) is a spontaneous disease characterised by repeated episodes of intraocular inflammation. The epidemiology of ERU has not been fully elucidated, but the condition appears to be much more common in horses than is recurrent uveitis in humans, especially in certain breeds and geographical regions. Both humans and horses show a similarly altered immune response and a marked autoimmune response as the primary disease pathophysiology. However, an inciting cause is not always clear. Potential inciting factors in horses include microbial agents such as Leptospira spp. Microbial factors and genetic predisposition to the disease may provide clues as to why the horse appears so susceptible to this disease. The aim of this review is to discuss the immunology and genetics of ERU, compare the disease in horses with autoimmune anterior uveitis in humans, and discuss potential reasons for the increased prevalence in the horse., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Leptospirosis is a zoonotic disease of global importance, and has a worldwide distribution. Equine leptospirosis is commonly manifested by recurrent uveitis, reproductive disorders, as abortions, embryonic absorption, stillbirth and the birth of weak foals. The aim of this study was to verify the presence of Leptospira sp or its DNA in genital tract of mares with reproductive problems. A total of 38 mares with reproductive problems were studied. All the mares were sampled for blood (for serology), urine (for culturing and qPCR), vaginal fluid-VF and endometrial biopsy-EB (for culturing, qPCR and indirect immunofluorescence). PCRs products were sequenced for secY gene. Seventeen (44.7%) serum samples were reactive, predominantly against serogroups Australis (76.4%) and Pomona (23.6%). No positive culture was obtained, but DNA was detected by qPCR on urine samples (26.3%), VF (44.7%) and EB (18.4%) collected 2 months or longer following diagnosis of early fetal death and endometritis. Leptospira cell aggregations were visible by indirect immunofluorescence on 57.1% (4/7) EBs and 17.6% (3/17) VFs. A total of 18 amplicons showed interpretable sequences. Out of those 18 amplicons, 15 presented 100% of identity with the species L. interrogans (sv Bratislava and Pomona), while three were L. borgpertersenii. This study suggests the presence of leptospires in the uterus of mares with reproductive problems. Moreover, serology was shown not to be indicated for the diagnosis of presumptive Leptospira infection in early gestation. The most common agent of the genital infection in those mares was L. interrogans, most probably sg Australis., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Retinal pigment epithelium (RPE) builds the outer blood-retinal barrier of the eye and plays an important role in pathogenesis of the sight threatening disease equine recurrent uveitis (ERU). ERU is a spontaneous autoimmune mediated inflammatory disease characterised by the breakdown of the outer blood-retinal barrier and an influx of autoaggressive T-cells into the inner eye. Therefore, identification of molecular mechanisms contributing to changed function of blood-retinal barrier in ERU is important for the understanding of pathophysiology. Cell surface proteins of RPE collected from healthy horses and horses with ERU were captured by in situ biotinylation and analysed with high resolution mass spectrometry coupled to liquid chromatography (LC-MS/MS) to identify differentially expressed proteins. With label free differential proteomics, a total of 27 differently expressed cell surface proteins in diseased RPE could be detected. Significant down-regulation of three very interesting proteins, synaptotagmin 1, basigin and collectrin was verified and further characterised., Biological Significance: We applied an innovative and successful method to detect changes in the plasma cell surface proteome of RPE cells in a spontaneous inflammatory eye disease, serving as a valuable model for human autoimmune uveitis. We were able to identify 27 differentially expressed plasma cell membrane proteins, including synaptotagmin 1, basigin and collectrin, which play important roles in cell adhesion, transport and cell communication., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Two horses with chronic uveitis and histological lesions consistent with equine recurrent uveitis (ERU) were examined. Microscopical findings in the ciliary body included deposits of amyloid lining the non-pigmented epithelium, intracytoplasmic, rod-shaped, eosinophilic inclusions and intraepithelial infiltration of T lymphocytes. Ultrastructural examination of the ciliary body of one horse confirmed the presence of abundant extracellular deposits of non-branching fibrils (9-11 nm in diameter) consistent with amyloid. Immunohistochemistry revealed strong positive labelling for AA amyloid and mass spectrometry showed the amyloid to consist primarily of serum amyloid A1 in both cases. The findings suggest that localized, intraocular AA amyloidosis may occur in horses with ERU., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Appaloosa horses are predisposed to equine recurrent uveitis (ERU), an immune-mediated disease characterized by recurring inflammation of the uveal tract in the eye, which is the leading cause of blindness in horses. Nine genetic markers from the ECA1 region responsible for the spotted coat color of Appaloosa horses, and 13 microsatellites spanning the equine major histocompatibility complex (ELA) on ECA20, were evaluated for association with ERU in a group of 53 Appaloosa ERU cases and 43 healthy Appaloosa controls. Three markers were significantly associated (corrected P-value <0.05): a SNP within intron 11 of the TRPM1 gene on ECA1, an ELA class I microsatellite located near the boundary of the ELA class III and class II regions and an ELA class II microsatellite located in intron 1 of the DRA gene. Association between these three genetic markers and the ERU phenotype was confirmed in a second population of 24 insidious ERU Appaloosa cases and 16 Appaloosa controls. The relative odds of being an ERU case for each allele of these three markers were estimated by fitting a logistic mixed model with each of the associated markers independently and with all three markers simultaneously. The risk model using these markers classified ~80% of ERU cases and 75% of controls in the second population as moderate or high risk, and low risk respectively. Future studies to refine the associations at ECA1 and ELA loci and identify functional variants could uncover alleles conferring susceptibility to ERU in Appaloosa horses., (© 2014 Stichting International Foundation for Animal Genetics.)

Equine recurrent uveitis is a spontaneous, lymphocyte-driven autoimmune disease. It affects horses worldwide and presents with painful remitting-relapsing inflammatory attacks of inner eye structures eventually leading to blindness. Since lymphocytes are the key players in equine recurrent uveitis, we were interested in potential changes of their protein repertoire which may be involved in disease pathogenesis. To create a reference for differential proteome analysis, we first unraveled the equine lymphocyte proteome by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subsequently identified 352 protein spots. Next, we compared lymphocytes from ERU cases and healthy horses with a two-dimensional fluorescence difference in gel electrophoresis approach. With this technique, we identified seven differentially expressed proteins between conditions. One of the significantly lower expressed candidates, septin 7, plays a role in regulation of cell shape, motility and migration. Further analyses revealed T cells as the main cell type with decreased septin 7 abundance in equine recurrent uveitis. These findings point to a possible pathogenetic role of septin 7 in this sight-threatening disease.