The refractive state of the eye in Icelandic horses with the Silver mutation.

BACKGROUND: The syndrome Multiple Congenital Ocular Anomalies (MCOA) is a congenital eye disorder in horses. Both the MCOA syndrome and the Silver coat colour in horses are caused by the same missense mutation in the premelanosome protein (PMEL) gene. Horses homozygous for the Silver mutation (TT) are affected by multiple ocular defects causing visual impairment or blindness. Horses heterozygous for the Silver mutation (CT) have less severe clinical signs, usually cysts arising from the ciliary body iris or retina temporally. It is still unknown if the vision is impaired in horses heterozygous for the Silver mutation. A recent study reported that Comtois horses carrying the Silver mutation had significantly deeper anterior chambers of the eye compared to wild-type horses. This could potentially cause refractive errors. The purpose of the present study was to investigate if Icelandic horses with the Silver mutation have refractive errors compared to wild-type horses. One hundred and fifty-two Icelandic horses were included in the study, 71 CT horses and five TT horses. All horses were genotyped for the missense mutation in PMEL. Each CT and TT horse was matched by a wild-type (CC) horse of the same age ± 1 year. Skiascopy and a brief ophthalmic examination were performed in all horses. Association between refraction and age, eye, genotype and sex was tested by linear mixed-effect model analysis. TT horses with controls were not included in the statistical analyses as they were too few. RESULTS: The interaction between age and genotype had a significant impact on the refractive state (P = 0.0001). CT horses older than 16 years were on average more myopic than wild-type horses of the same age. No difference in the refractive state could be observed between genotypes (CT and CC) in horses younger than 16 years. TT horses were myopic (-2 D or more) in one or both eyes regardless of age. CONCLUSION: Our results indicate that an elderly Icelandic horse (older than 16 years) carrying the Silver mutation is more likely to be myopic than a wild-type horse of the same age.

Initial Impact of Different Feeding Methods on Feed Intake Time in Stabled Icelandic Horses.

The natural behaviour of horses is to spend the majority of their time on feed intake The feeding of stabled horses is, however, often far from that, as their feed intake is limited to their nutritional requirements. In order to approach their natural foraging time, it is important to extend the feed intake time of stabled horses. The aim of this study was to estimate if the feed intake time differs when feeding haylage in a haynet, hayball, metal corner manger, or from the box floor. The experimental design consisted of a Latin square, occurred across four days with four adult Icelandic horses and four treatments. Horses were stabled in individual boxes and fed 7 kg of high-energy haylage in two even meals while the intake time was recorded. The feed intake time per kg DM was shorter from the manger or the box floor than from a haynet or hayball (81 or 85 min versus 94 or 96 min; p < 0.05). It can be concluded that feeding haylage in a hayball or in a haynet can increase the feed intake time by 13% per day (12 min/kg DM/day) when compared to the more traditional methods. Thus, with simple alternatives, it is possible to extend the feed intake time of stabled horses.

In vitro induction of functional allergen-specific CD4+ CD25high Treg cells in horses affected with insect bite hypersensitivity.

BACKGROUND: Insect bite hypersensitivity (IBH) is a recurrent allergic dermatitis of horses with similarities to human atopic eczema, caused by bites of insects of the genus Culicoides. Previous studies suggested a dysregulated T cell tolerance to Culicoides allergen in IBH-affected horses. OBJECTIVE: We have investigated whether the suppressive function of CD4(+) CD25(high) cells is impaired in IBH-affected horses and possible ways to restore it. METHODS: CD4(+) CD25(-) cells sorted from peripheral blood mononuclear cells (PBMC) were stimulated with irradiated autologous PBMC pulsed with Culicoides or tetanus toxoid as control antigen, in the presence of CD4(+) CD25(high) cells. Furthermore, Culicoides-specific CD4(+) CD25(high) regulatory cells were expanded or induced from CD4(+) CD25(-) cells in vitro in the presence of a combination of rIL-2 and rTGF-ß1 (rIL-2/rTGF-ß1) or of retinoic acid and rapamycin (RetA/Rapa). Proliferation was determined by [(3) H] thymidine incorporation and cytokine production measured by flow cytometry. RESULTS: The ability of Culicoides- but not tetanus-stimulated CD4(+) CD25(high) cells to suppress proliferation of CD4(+) CD25(-) cells was significantly lower in IBH-affected horses (28%) than in healthy controls (86%). The decreased suppression in IBH-affected horses was associated with a significantly higher proportion of IL-4(+) cells and a lower percentage of FoxP3(+) IL-10(+) compared to controls. Addition of rIL-2/rTGF-ß1 or of RetA/Rapa to Culicoides-stimulated CD4(+) CD25(high) cells from IBH-affected horses significantly increased the proportion of FoxP3(+) IL-10(+) cells. We also found that RetA/Rapa induced a more significant decrease in the frequency of IL-4(+) cells than rIL-2/rTGF-ß1. Moreover, the suppressive activity of Culicoides-stimulated CD4(+) CD25(high) cells was significantly restored by both rIL-2/rTGF-ß1and RetA/Rapa, albeit in an antigen-unspecific manner. In contrast, in vitro induced Culicoides-specific CD4(+) CD25(high) cells suppressed proliferation of CD4(+) CD25(-) cells in an antigen-specific manner. CONCLUSION AND CLINICAL RELEVANCE: The in vitro induction of functional allergen-specific Treg cells in IBH-affected horses suggests a potential therapeutic use of these cells in allergy.

The Frequency of Errors in Determining Age Based on Selected Features of the Incisors of Icelandic Horses.

The structure and changes occurring to horses' teeth during ontogeny are not only used to assess the degree of somatic maturity but also the development of universal patterns and is therefore used to determine the age of horses. Research shows that methods of assessing the age of horses based on the appearance of teeth tend to suffer from relatively large errors. This is probably influenced by the results of intensive selection and being kept in living conditions that differ substantially from their natural environment. This study aimed to assess the suitability of selected features of the incisors to determine the age of Icelandic horses. One hundred and twenty-six Icelandic horses (78 mares and 48 stallions) of different ages (range: 0-24 years; groups: 0-2 years, >2-5 years, >5-11 years, and >11 years) were examined by an experienced horse person who was blinded to the actual age of the horse and did not know which age group horses were in. Age was determined by the inspection of each horse's teeth and was compared to the actual age of the horse recorded in the breeding documentation, and the percentage of mistakes made regarding the age group was calculated. The estimated age did not match the real age in 36.5% of the horses. The age was more often underestimated (19.0%) by, on average, 0.9 ± 1.0 years than overestimated (17.5%) by, on average, 1.3 ± 1.4 years. Within age groups, the least number of errors in determining age were made in young horses aged 0-2 years, when the eruption and growing of the deciduous incisors and the disappearance of the cups was considered. The average percentage of errors in this group (2.1%) was significantly lower (p < 0.01) than for older horses, whose age was estimated based on the exchange of deciduous to permanent teeth (55.8%), disappearance of the cups (68.0%), and shape changes on the grinding surface (40.0%). Significantly more frequent underestimation of age based on replacing deciduous for permanent incisors and significantly more frequent overestimation of age on the basis of the disappearance of the cup may indicate that Icelandic horses up to 5 years of age are characterized by a slower rate of growth than horses of other breeds, especially warmblood horses. These results suggest that patterns used to determine the real age of horses based on changes occurring on the incisors should be modified in order to consider the specificity of the course of growth and maturation processes of horses of various types and breeds.

A prospective study on insect bite hypersensitivity in horses exported from Iceland into Switzerland.

BACKGROUND: Insect bite hypersensitivity (IBH) is an IgE-mediated dermatitis caused by bites of Culicoides spp., which occurs frequently in horses imported from Iceland to continental Europe. IBH does not occur in Iceland because Culicoides species that bite horses are not present. However, Simulium vittatum (S. vittatum) are found in Iceland. In Europe, blood basophils from IBH-affected horses release significantly more sulfidoleukotrienes (sLT) than those from healthy controls after in vitro stimulation with Culicoides nubeculosus (C. nubeculosus) and S. vittatum. Aims of the study were: (I) using the sLT release assay, to test if horses living in Iceland were sensitized to S. vittatum and (II) to determine in a longitudinal study in horses imported from Iceland to Switzerland whether the sLT release assay would allow to predict which horses would develop IBH. RESULTS: Horses in Iceland, even when living in high S. vittatum areas, were usually not sensitized to S. vittatum or C. nubeculosus. Incidence of IBH in the 145 horses from the longitudinal study was 51% and mean time until IBH developed was 2.5 ± 1 year. Before import and after the first summer following import, there were no significant differences in sLT release between the endpoint healthy (H) and IBH groups. After the 2nd summer, when the number of clinically affected horses increased in the endpoint IBH group, a significantly higher sLT release after stimulation with C. nubeculosus but not with S. vittatum was observed. After the 3rd and 4th summer, the endpoint IBH group had a significantly higher sLT release with C. nubeculosus and S. vittatum than the endpoint H group. Some of the horses that remained healthy became transiently positive in the sLT release assay upon stimulation of their peripheral blood leucocytes with C. nubeculosus. CONCLUSIONS: Horses in Iceland are not sensitized to S. vittatum. In horses that develop IBH, sensitization to S. vittatum is secondary to sensitization to C. nubeculosus and probably a result of an immunological cross-reactivity. A sLT release assay cannot be used to predict which horses will develop IBH. A transient positive reaction in the sLT release assay observed in horses that remained healthy suggests that immunoregulatory mechanisms may control an initial sensitization of the healthy horses.