Ocular lesions of bovine malignant catarrhal fever

The ocular lesions of bovine malignant catarrhal fever were characterized in 15 naturally occurring and eight experimentally induced cases of the disease. Consistent findings included: lymphocytic vasculitis of retinal, scleral, posterior ciliary, and uveal vessels; uveitis, especially involving ciliary processes, ciliary body, and iris; and keratitis with corneal edema, neovascularization, and epithelial and endothelial degeneration. Lymphocytic ciliary neuritis and optic meningitis were found less frequently. Ultrastructural examination of the ciliary body and iris from one experimental calf confirmed that most infiltrating mononuclear cells were lymphocytes. The uveitis, vasculitis, and keratitis of malignant catarrhal fever were probably immune-mediated.     

Depiction of the structure of the vitreous body in horses without ocular diseases and in horses with equine recurrent uveitis (ERU) using transmission electron microscopy

Neither the ultrastructure of the vitreous body from horses without ocular diseases, nor the pathomorphological changes in the vitreous body associated with equine recurrent uveitis (ERU) have been described. However, the vitreous body plays an important role in the pathogenesis of ERU. Ten vitreous body samples obtained from 5 horses without ocular disease, and 38 vitreous body samples from horses with ERU (collected during vitrectomy) were examined by transmission electron microscopy. The vitreous body samples of horses without ocular diseases were characterized by a loose network of unbranched fibrils 10-12 nm in width. In the vitreous body samples of horses with ERU numerous dense bundles of fibrils, mononuclear inflammatory cells and necrotic cells represent the destruction of the vitreous fibrillar network. In this study, equine vitreous body ultrastructure was described for the first time. Thus, demonstrating ultramorphologically, the clinically apparent changes of the vitreous body associated with ERU.     

Granulomatous uveitis associated with vaccination in the atlantic salmon

This study addressed histologic and immunopathologic changes in ocular tissues and investigated the distribution of major histocompatibility class II (MHC class II)-positive cells in Atlantic salmon (Salmo salar) suffering from severe postvaccination disease. Twenty-nine fish with generalized inflammation, probably a result of vaccination, were investigated. One individual that had escaped vaccination was included in the study. Material was investigated by cultivation methods for fungi and bacteria. Histology using conventional staining procedures and immunohistochemistry with antisera against MHC class II beta chain were performed. No growth was observed from the cultivation investigations. Histology revealed occlusion of the lumen in the larger choroid vessels and in the choriocapillaris, inflammatory infiltrations and loss of structure in the choroid rete, and, in some cases, aggregations of multinucleated giant cells (MGC) and Splendore-Hoeppli material. Immunohistochemistry demonstrated massive MHC class II+ cellular infiltrations in the uveal tract. Such infiltrations were also seen in the ventral ciliary cleft, a condition that is associated with glaucoma. Immunoreactive cells included dendritelike cells, epithelioid cells, and MGCs. The endothelia of smaller vessels were frequently MHC class II+, and immunoreactive infiltrations were seen in the optic nerve in several individuals. No pathologic changes were detected in the unvaccinated individual. In conclusion, generalized inflammatory reactions in fish may lead to severe ocular inflammation, occlusion of uveal vessels, and perivascular changes with MHC class II+ upregulation in cells in the uveal tract and optic nerve.     

Surgical management of equine recurrent uveitis with single port pars plana vitrectomy

Current information suggests that equine recurrent uveitis (ERU) is an immune-mediated reaction to infectious agents or to autologous ophthalmic tissue. Recurrences are associated with progression of irreversible ocular damage. This report describes the intraoperative technique, complications, and long-term results of 38 eyes in 35 horses with ERU that underwent pars plana vitrectomy. The majority of the horses were warm-blooded. Recurrence of ERU was prevented in 35 of the 38 eyes. Some horses, especially in patients with incipient cataracts, developed vision loss in postoperative, quiescent eyes which was usually associated with cataract formation. Vision was stable in 85% of all eyes that underwent vitrectomy. Pars plana vitrectomy in horses appears successful in interrupting the cycle of repeated episodes of ERU, and the subsequent globe destruction in the majority of eyes. Removal of uveitis-induced 'immunologic memory' in the vitreous by vitrectomy may reduce adverse interaction between the vitreous and the uveal tract, and therefore reduce the recurrence of ERU.     

Immunophenotypic analysis of the distribution of hepatic macrophages,lymphocytes and hepatic stellate cells in the adult rat liver

The liver consists of parenchymal hepatocytes and non-parenchymal cells. Non-parenchymal cells, Kupffer cells, hepatic stellate cells and cholangiocytes have crucial roles in liver homeostasis and liver pathology. To establish baseline data, this study investigated immunohistochemically the distribution of non-parenchymal cells in perivenular areas (PV), periportal areas (PP) and Glisson's sheath (GS) of adult rat liver. Liver tissues were collected from the left lateral lobe of rats. CD163-positive macrophages were seen along the sinusoid of PV and PP areas, indicating Kupffer cells. Double immunofluorescence showed, Kupffer cells partly co-expressed CD68 and MHC class II antigens in the liver. The numbers of Kupffer cells were significantly high in PP areas as compared with PV or GS areas. CD68-positive exudative macrophages were highly localized in PP and GS areas and a comparatively low PV area. MHC class II-positive dendritic cells (activated macrophages) were localized mainly in GS. Granzyme B-positive NK cells were mainly localized in the Glisson's sheath. CD3-positive T cells and CD20-positive B cells were distributed along the sinusoids of the PP and PV areas of hepatic lobules. Vimentin and glial fibrillary acidic protein (GFAP)-positive hepatic stellate cells were localized along sinusoids in the hepatic lobules of the liver. Cholangiocytes reacting to cytokeratin 19 were seen on interlobular bile ducts in Glisson's sheath of the liver. This study shows that heterogeneous macrophage populations, liver-resident lymphocytes and hepatic stellate cells localized in PP and PV areas or GS areas of the liver with cells specific patterns.     

Characterization of cytokines associated with Th17 cells in the eyes of horses with recurrent uveitis

Objective Equine recurrent uveitis (ERU) is a spontaneous disease that is the most common cause of blindness in horses, affecting up to 15% of the horse population. Th17 cells are a major cell population driving the pathogenesis in several mouse models of autoimmune inflammation, including experimental autoimmune uveitis. The purpose of this study is to investigate the role a Th17 cell‐mediated response plays in the pathogenesis of ERU. Procedure Banked, Davidson’s‐fixed equine globes histopathologically diagnosed with ERU (n = 7) were compared immunohistochemically with healthy control globes (n = 7). Immunohistochemical staining was performed using a pan‐Leptospira antibody and antibodies against IL‐6, IL‐17, and IL‐23. Additionally, immunostaining was performed for T‐cell (CD3) and B‐cell (CD79α) markers. Specificity of immunoreactivity was confirmed by western blot analysis. Results Immunohistochemical staining was positive for IL‐6, IL‐17, and IL‐23 within the cytoplasm of nonpigmented ciliary epithelial cells and mononuclear inflammatory cells infiltrating the iris, and ciliary body of ERU horses (n = 7) but negative in controls (n = 7). ERU‐affected eyes were CD3 positive (n = 7) and CD79α negative (n = 7). Staining for Leptospira was negative in all ERU and control globes. Conclusions Strong immunoreactivity for IL‐6, IL‐17, and IL‐23, in conjunction with the fact that T lymphocytes are the predominating inflammatory cells present in ERU, suggests that IL‐17‐secreting helper T‐cells play a role in the pathogenesis of ERU. These findings suggest that horses with ERU may serve as a naturally occurring animal model for autoimmune uveitis.     

Detection of Leptospira interrogans DNA and antigen in fixed equine eyes affected with end-stage equine recurrent uveitis.

Equine recurrent uveitis (ERU) is the most frequent cause of blindness in horses worldwide. Leptospira has been implicated as an etiologic agent in some cases of ERU and has been detected in fresh ocular tissues of affected horses. The objective of this study was to determine the presence of Leptospira antigen and DNA in fixed equine ocular tissues affected with end-stage ERU. Sections of eyes from 30 horses were obtained. Controls included 1) 10 normal equine eyes and 2) 10 equine eyes with a nonrecurrent form of uveitis. The experimental group consisted of 10 eyes diagnosed with ERU based on clinical signs and histologic lesions. Sections were subjected to immunohistochemical staining with an array of rabbit anti-Leptospira polyclonal antibodies. DNA extractions were performed by using a commercial kit designed for fixed tissue. Real-time PCR analysis was completed on extracted DNA. The target sequence for PCR was designed from alignments of available Leptospira 16S rDNA partial sequences obtained from GenBank. Two of 10 test samples were positive for Leptospira antigen by immunohistochemical assay. Zero of 20 controls were positive for Leptospira antigen. All test samples and controls were negative for Leptospira DNA by real-time PCR analysis. Leptospira was detected at a lower frequency than that previously reported for fresh ERU-affected aqueous humor and vitreous samples. Leptospira is not frequently detectable in fixed ocular tissues of horses affected with ERU when using traditional immunohistochemical and real-time PCR techniques.     

Diagnosis of Borrelia‐associated uveitis in two horses

Borrelia burgdorferi, the etiologic agent of Lyme disease is a tick born spirochetal infection. Clinical signs of Lyme borreliosis are uncommon in horses, but when present they are often vague and nonspecific. In horses, Lyme borreliosis has been implicated in musculoskeletal, neurological, reproductive, and ocular disorders, including uveitis, but definitive diagnosis can be challenging as the causative agent is rarely isolated and serologic tests can be unreliable and do not confirm active disease. Here, we report two cases of equine uveitis associated with B. burgdorferi based on the identification of spirochetes within ocular fluids and confirmed with PCR testing. The two cases illustrate some of the challenges encountered in the recognition and diagnosis of equine Lyme borreliosis. Although only one of many possible causes of equine uveitis, Lyme disease should be considered a differential diagnosis, especially in endemic areas. Given the possibility for false negative results of serum tests during uveitis associated with B. burgdorferi and the failure of such tests to confirm active infection, a combination of cytologic assessment, antibody, and/or PCR testing of ocular fluids may be worthwhile if the clinical suspicion for Lyme uveitis is high.     

The pathogenesis and significance of pre-iridal fibrovascular membrane in domestic animals

Histologic examination was made of 1,419 globes from domestic animals (964 dogs, 374 cats, 41 horses, and 40 cattle) with ocular disease; pre-iridal membranes (rubeosis iridis) were found in 98. The membranes originated as endothelial budding from the anterior iridal stroma and seemed to mature into fibrous or fibrovascular membranes that were often followed by hyphema or, occasionally, glaucoma. Pre-existent disease in the 98 affected globes included chronic endophthalmitis (27/98), chronic glaucoma (24/98), anterior uveal melanoma (15/98), ciliary body adenoma (14/98), neoplasms metastatic to the eye (8/98), and chronic retinal detachment (6/98). In terms of likelihood of occurrence, pre-iridal membranes seen in 21% (6/21) of globes with retinal detachment, 19% (14/75) of those with ciliary body adenomas, 14% (24/167) of those with chronic glaucoma, and 10% (15/158) of those with anterior uveal melanoma. They were detected with greatest relative frequency in horses (9/41) followed by dogs (83/964), cats (5/374) and cattle (1/40). These membranes, which are rarely detected by clinical examination, probably form in response to angiogenic factors released by ischemic retina, by neoplasms, or by leukocytes involved in ocular inflammation.     

Serological study of leptospiral infections and endogenous uveitis among horses and ponies in the United Kingdom

The prevalence of antibody titres to a range of 20 leptospira antigens in the serum of horses and ponies with no ophthalmic abnormalities and with ophthalmoscopic evidence of endogenous uveal inflammatory disease was determined using a microscopic agglutination technique. Titres against leptospira antigens were observed in 13 out of 138 (9.1 per cent) animals with no ophthalmic abnormalities, and in three out of 27 (11.1 per cent) animals with anterior uveitis. Serovar sejroe was common to all seropositive animals with anterior uveitis. The results show that leptospira infection is not a major factor in the aetiology of equine anterior uveitis in the UK, but suggests that the organism may be associated with some cases of recurrent anterior uveitis.     

Normal structure and age-related changes of the equine retina

Investigations of the pathophysiology of ocular diseases require a detailed knowledge of the microanatomy of the eye. The available information is still inadequate for the equine retina despite the importance of eye diseases in equine medicine. Here we provide a comprehensive analysis of the histologic features of the horse eye as a reference for future studies. Thirty normal eyes of 15 healthy horses were examined immediately after slaughter. The retina of the horse differs considerably in the degree and quantity of neurons and glial elements as well as in vascular patterns compared to the retina of other domestic animals. Morphometric analysis revealed that the thickness of the retina varies between 80 microm at the ora serrata and 250 microm medial to the optic disc. Approximately 90% of the equine retina is comparatively thin (< 130 microm). This is a physiologic response to the distance that oxygen can diffuse in avascular retina. Ganglion cells form a single layer in all parts of the retina. The majority of ganglion cells are very large Nissl-positive cells. Small Nissl-negative ganglion cells are less abundant. A high ganglion cell density is found only in the central area. Vascularization is virtually absent from the retina with the exception of a narrow strip around the disc of the optic nerve, as revealed by lectin histochemistry. Light microscopy of the eyes of older horses repeatedly revealed cystoid degenerations in the retina adjacent to the pars plana of the ciliary body, as well as a destruction of the regular layering of the peripheral region of the retina.     

Flow cytometric analysis of blood lymphocyte phenotypes in horses infected with the equine infectious anemia virus

Lymphocyte phenotypes were evaluated in bloodsamples taken from horses in the persistent phase EIA virus infection (n=10), from diseased controls (n=5) and from normal controls (n=10). A single animal in the acute phase of EIA was also studied. Cells were identified using flow cytometry after labelling with polyclonal antibodies to horses immunoglobulins for B-lymphocytes, or monoclonal antibodies (MAbs) to CD4, CD5, CD8 and MHC Class-II antigens. In horses persistently infected with EIA virus, the percentage of CD4+T-lymphocytes is systematically reduced and the percentage of CD8+T-lymphocytes is irregular, ranging from normal to severely reduced. Most of them have low values for cells expressing class-II antigens, but high B-cell percentages. Total CD5+ cell percentages are low in all diseased horses examined compared to normal controls. The acutely-infected foal differed from the persistently infected animals in having an elevated percentage of CD8+ T-lymphocytes, and a severely reduced percentage of B-cells.BSA, bovine serum albumin; EIA, equine infectiousanemia; FITC, fluorescein isothiocyanate; MAb, monoclonal antibody; PBL, peripheral blood lymphocytes; PBMC, peripheral blood mononuclear cells, PBS, phosphate buffered saline.     

Concurrent clinical intraocular findings in horses with depigmented punctate chorioretinal foci

Objective To report concurrent clinical intraocular findings in horses with depigmented punctate chorioretinal foci and to document any correlation with equine recurrent uveitis (ERU). Procedure Records of 131 horses (241 eyes) examined at the University of Georgia Veterinary Teaching hospital from 2001 to 2010 were reviewed with either clinically normal fundi or depigmented punctate chorioretinal foci in the absence of other fundic pathology. Data collected included patient signalment, concurrent clinical ocular findings and follow‐up information. Sex, presence of no other intraocular findings, presence of ERU, presence of cataracts, and presence of vitreal disease were compared between normal and foci groups using chi‐squared analysis. Age and length of follow‐up time were compared using a student’s t‐test. Results Ninety‐one horses (167 eyes) with chorioretinal foci and forty horses (74 eyes) with clinically normal ocular fundi were examined. Fifty‐eight (64%) horses with chorioretinal foci and 20 (50%) horses with clinically normal fundi had a normal intraocular examination. There was no significant difference in any of the criteria examined between groups. Conclusions Horses with depigmented punctate chorioretinal foci, in the absence of other fundic pathology, are not more likely to have intraocular disease or ERU than horses with clinically normal ocular fundi. These findings suggest that depigmented punctate fundic foci in horses are not indicative of or associated with ERU.     

Feline uveitis: diagnosis and treatment

Uveitis is the inflammation of any or all parts of the vascular tunic of the eye; the vascular tunic includes the iris, the ciliary body, and choroid. A good knowledge base, up-to-date reference materials, and good instruments will improve the diagnosis of uveitis. Feline uveitis can be caused by numerous infectious agents in addition to neoplasia and less likely trauma. The infectious causes most commonly associated with feline uveitis include feline leukemia virus, feline immunodeficiency virus, feline infectious peritonitis, systemic fungal infections, toxoplasmosis, and bartonellosis. Neoplastic causes of uveitis can be primary or secondary. Iris melanoma is the most common primary uveal neoplasia and trauma-associated sarcoma is the second most common primary uveal neoplasia. Treatment for the clinical signs of anterior uveitis include topical steroidal or non-steroidal anti-inflammatory agents, parasympatholytic agents for ciliary spasm, to keep the pupil dilated, and to prevent posterior synechia. Posterior uveitis should be treated with systemic medications that will address the underlying cause. Enucleation of blind, painful eyes not responsive to medications is a means to alleviate the animal's discomfort and to further diagnose the underlying cause.     

Immunohistochemical investigation of the distribution of immunoglobulins G, A and M within the anterior uvea of the normal equine eye

Distribution of the immunoglobulin (Ig) classes G, A and M within the anterior uvea of eight clinically normal equine eyes was examined using indirect immunoperoxidase labelling. Increased staining intensity of stromal IgG and IgA was observed within the ciliary processes, the iris stroma being relatively free of immunoglobulin. This may reflect anatomical variation in the permeability of the uveal microvasculature to lipid insoluble plasma macro-molecules. Intracellular IgG and IgA were observed within the non-pigmented ciliary epithelium in seven and four of the eight eyes respectively, although wide variation between numbers of Ig bearing cells in each eye was noted. It was suggested that these cells may have a role in the removal of Ig from the posterior chamber aqueous. Plasma cells, of IgG isotype, were observed in only one eye, suggesting that intraocular production of Ig is not a feature of the normal equine eye.     

The long‐term effects of semiconductor diode laser transscleral cyclophotocoagulation on the normal equine eye and intraocular pressurea

Objective To describe the long‐term histologic and intraocular pressure (IOP) lowering effects of diode laser transscleral cyclophotocoagulation (TSCP) on the normal equine eye. Animals Eight normal adult horses. Procedures TSCP was performed in one randomly assigned eye. Sixty spots were treated at settings of 1500 ms and 1500 mW. Two horses were randomly selected for euthanasia at 2, 4, 12, or 24 weeks post‐TSCP. Both eyes were enucleated and histologically evaluated. Intraocular pressure was measured by applanation tonometry prior to TSCP, immediately post‐TSCP, twice daily for 7 days post‐TSCP and then monthly until study conclusion. A longitudinal model estimated the average IOP values for the treated and untreated eyes at 1 week, 1, 3, and 6 months post‐TSCP. Results All treated eyes at all time periods exhibited four characteristic histologic lesions: scleral collagen hyalinization, ciliary body pigment dispersion and clumping, focal disruption of the ciliary body epithelium, and focal ciliary process atrophy. After TSCP, there were no significant changes in IOP from baseline for the control eyes, while the IOP in treated eyes was significantly decreased from baseline (P < 0.05) at all time periods. The estimated decrease in IOP in the treated eyes compared to baseline IOP at 6 months was ‐3.76 mmHg for an average decrease in IOP of 20% from baseline. Conclusion  Diode laser TSCP produces histologic lesions in the equine ciliary body that result in a significant and sustained decrease in IOP. TSCP may be an effective management for equine glaucoma.