Vincent Gerber

Mutations in MITF and PAX3 cause "splashed white" and other white spotting phenotypes in horses.

During fetal development neural-crest-derived melanoblasts migrate across the entire body surface and differentiate into melanocytes, the pigment-producing cells. Alterations in this precisely regulated process can lead to white spotting patterns. White spotting patterns in horses are a complex trait with a large phenotypic variance ranging from minimal white markings up to completely white horses. The “splashed white” pattern is primarily characterized by an extremely large blaze, often accompanied by extended white markings at the distal limbs and blue eyes. Some, but not all, splashed white horses are deaf. We analyzed a Quarter Horse family segregating for the splashed white coat color. Genome-wide linkage analysis in 31 horses gave a positive LOD score of 1.6 in a region on chromosome 6 containing the PAX3 gene. However, the linkage data were not in agreement with a monogenic inheritance of a single fully penetrant mutation. We sequenced the PAX3 gene and identified a missense mutation in some, but not all, splashed white Quarter Horses. Genome-wide association analysis indicated a potential second signal near MITF. We therefore sequenced the MITF gene and found a 10 bp insertion in the melanocyte-specific promoter. The MITF promoter variant was present in some splashed white Quarter Horses from the studied family, but also in splashed white horses from other horse breeds. Finally, we identified two additional non-synonymous mutations in the MITF gene in unrelated horses with white spotting phenotypes. Thus, several independent mutations in MITF and PAX3 together with known variants in the EDNRB and KIT genes explain a large proportion of horses with the more extreme white spotting phenotypes.

Effects of genetic and environmental factors on chronic lower airway disease in horses

BACKGROUND Environment and genetics influence the manifestation of recurrent airway obstruction (RAO), but the associations of specific factors with mild, moderate, and severe clinical signs are unknown. HYPOTHESIS We hypothesized that sire, feed, bedding, time outdoors, sex, and age are associated with clinical manifestations of mild, moderate, and severe lower airway disease. ANIMALS Direct offspring of 2 RAO-affected Warmblood stallions (F1S1, n = 172; F1S2, n = 135); maternal half-siblings of F1S1 (mHSS1, n = 66); and an age-matched, randomly chosen control group (CG, n = 33). METHODS A standardized questionnaire was used to assess potential risk factors and to establish a horse owner assessed respiratory signs index (HOARSI 1-4, from healthy to severe) according to clinical signs of lower airway disease. RESULTS More F1S1 and F1S2 horses showed moderate to severe clinical signs (HOARSI 3 and HOARSI 4 combined, 29.6 and 27.3%, respectively) compared with CG and mHSS1 horses (9.1 and 6.2%, respectively; contingency table overall test, P < .001). Sire, hay feeding, and age (in decreasing order of strength) were associated with more severe clinical signs (higher HOARSI), more frequent coughing, and nasal discharge. CONCLUSIONS AND CLINICAL RELEVANCE There is a genetic predisposition and lesser but also marked effects of hay feeding and age on the manifestation of moderate to severe clinical signs, most markedly on coughing frequency. In contrast, mild clinical signs were not associated with sire or hay feeding in our populations.

The Transcriptome of Equine Peripheral Blood Mononuclear Cells

Complete transcriptomic data at high resolution are available only for a few model organisms with medical importance. The gene structures of non-model organisms are mostly computationally predicted based on comparative genomics with other species. As a result, more than half of the horse gene models are known only by projection. Experimental data supporting these gene models are scarce. Moreover, most of the annotated equine genes are single-transcript genes. Utilizing RNA sequencing (RNA-seq) the experimental validation of predicted transcriptomes has become accessible at reasonable costs. To improve the horse genome annotation we performed RNA-seq on 561 samples of peripheral blood mononuclear cells (PBMCs) derived from 85 Warmblood horses. The mapped sequencing reads were used to build a new transcriptome assembly. The new assembly revealed many alternative isoforms associated to known genes or to those predicted by the Ensembl and/or Gnomon pipelines. We also identified 7,531 transcripts not associated with any horse gene annotated in public databases. Of these, 3,280 transcripts did not have a homologous match to any sequence deposited in the NCBI EST database suggesting horse specificity. The unknown transcripts were categorized as coding and noncoding based on predicted coding potential scores. Among them 230 transcripts had high coding potential score, at least 2 exons, and an open reading frame of at least 300 nt. We experimentally validated 9 new equine coding transcripts using RT-PCR and Sanger sequencing. Our results provide valuable detailed information on many transcripts yet to be annotated in the horse genome.

Clinical and echocardiographic features of mild mitral valve regurgitation in 108 horses

The clinical and echocardiographic characteristics of 108 horses with echocardiographically confirmed mild mitral valve regurgitation (MR) were investigated along with its clinical progression. Follow-up consisted of a re-examination of 28 horses and questionnaires were used to obtain information on a further 43 cases. Thirty-seven horses with mild MR were lost to follow-up. Horses with mild MR were re-examined between 2 and 9 years (3.8+/-1.8 years) following first presentation, with mild MR still present and a small, but statistically significant (P=0.049) increase of left ventricular diameter in end-diastole. These results suggested that mild MR has a good mid-term prognosis in sport and pleasure horses.

European outbreak of atypical myopathy in the autumn 2009.

BACKGROUND Atypical myopathy is an acute, severe rhabdomyolysis occurring in grazing horses. In the beginning of October 2009, a new outbreak occurred in several European countries. Geographic, demographic and clinical data of the reported cases in the month October 2009 are described. KEY FINDINGS The survival rate in this outbreak was 25%. The most frequently observed clinical signs were congested mucous membranes, dyspnea, tachycardia, depression, weakness, stiffness, recumbency, trembling, sweating, and myoglobinuria. Nonsurvivors were significantly more likely to be recumbent than survivors. Prognostic factors, symptomatic treatment, and preventive measures are discussed. SIGNIFICANCE Differences were encountered during the described outbreak of atypical myopathy in October 2009 compared with previous outbreaks reported. Equine practitioners should be aware that previous epidemiological studies have shown that after a high prevalence in the autumn, new cases are likely to occur in the following spring.

Haematological parameters are normal in dominant white Franches-Montagnes horses carrying a KIT mutation.

The KIT receptor protein-tyrosine kinase plays an important role during embryonic development. Activation of KIT is crucial for the development of various cell lineages such as melanoblasts, stem cells of the haematopoietic system, spermatogonia and intestinal cells of Cajal. In mice, many mutations in the Kit gene cause pigmentation disorders accompanied by pleiotropic effects on blood cells and male fertility. Previous work has demonstrated that dominant white Franches-Montagnes horses carry one copy of the KIT gene with the p.Y717X mutation. The targeted breeding of white horses would be ethically questionable if white horses were known to suffer from anaemia or leukopenia. The present study demonstrates that no statistically significant differences in peripheral blood parameters are detectable between dominant white and solid-coloured Franches-Montagnes horses. The data indicate that KIT mutations may have different effects in mice, pigs, and horses. The KIT p.Y717X mutation does not have a major negative effect on the haematopoietic system of dominant white horses.

Serological and clinical proof of freedom from Equine Infectious Anemia (EIA) in imported and domestic horses in Switzerland

Since 1991, no cases of Equine Infectious Anemia (EIA) have been reported in Switzerland. Risk factors for introduction of the virus into Switzerland are still present or have even increased as frequent inapparent infections, large numbers of imported horses, (since 2003) absence of compulsory testing prior to importation, EIA cases in surrounding Europe, possible illegal importation of horses, frequent short-term stays, poor knowledge of the disease among horse owners and even veterinarians. The aim of this study was to provide evidence of freedom from EIA in imported and domestic horses in Switzerland. The serum samples from 434 horses imported since 2003 as well as from 232 domestic horses fifteen years of age or older (since older horses have naturally had a longer time of being exposed to the risk of infection) were analysed using a commercially available ELISA test. All samples were seronegative, indicating that the maximum possible prevalence that could have been missed with this sample was 0.5% (95% confidence).

Equine Infektiöse Anämie (EIA)

Equine Infectious Anemia (EIA) is a reportable, eradicable epizootic disease caused by the equine lentivirus of the retrovirus family which affects equids only and occurs worldwide. The virus is transmitted by blood, mainly by sanguivorous insects. The main symptoms of the disease are pyrexia, apathy, loss of body condition and weight, anemia, edema and petechia. However, infected horses can also be inapparent carriers without any overt signs. The disease is diagnosed by serological tests like the Coggins test and ELISA tests. Presently, Switzerland is offi cially free from EIA. However, Switzerland is permanently at risk of introducing the virus as cases of EIA have recently been reported in different European countries.

Safety of Intra-Articular Gold Microimplants in Horses–A Randomized, Blinded, Controlled Experimental Study

Abstract Arthrogenic pain is a common problem in equids. Frequently used treatments such as systemic non‐steroidal anti‐inflammatory drugs or intra‐articular steroids can lead to severe side effects if used repeatedly. Gold has been used since ancient times to treat a variety of conditions and has anti‐inflammatory and immunomodulating properties. Results from clinical and in vitro studies suggest that local gold might provide a safe and effective alternative to alleviate articular pain. In particular, gold microimplants have been proposed to this end, but it is unknown if and how healthy joints react to this treatment. The aim of this study was to evaluate the safety of gold microimplants (Berlock‐Micro‐Implants) mixed with hyaluronic acid and injected into the middle carpal joint of nine healthy horses. Each horse was treated in one carpus and sham‐treated on the contralateral. Lameness, carpal temperature increase, swelling, and reactions to joint palpation were observed in both treated and sham‐treated carpi in the first week after needle arthroscopy and treatment. Significant differences between treated and sham‐treated carpi were found only for mechanical nociceptive thresholds 4 days after treatment. Higher thresholds were found in treated joints compared with sham‐treated joints. None of the outcome measures selected in the present study indicated systemic or local adverse effects specifically attributed to gold microimplants. In the absence of systemic and local adverse reactions to gold microimplants, the results of the present study support future clinical trials to test the pain‐relieving efficacy of this treatment modality in subacute or chronic articular inflammatory processes in equine patients. HighlightsThe safety of intra‐articular gold microimplants was evaluated in healthy horses.No systemic or local adverse effects specifically attributed to gold microimplants were found.The pain‐relieving efficacy of gold microimplants can now be evaluated in equine patients.

Evaluation of the effects of performance dentistry on equine rideability: a randomized, blinded, controlled trial

ABSTRACT Objective: This study attempted to determine: (1) if degree of dental malocclusion assigned prior to dental treatment was associated with equine rideability, assessed using a standardized score and (2) if performance dentistry improved this score. Animals: Thirty-eight Franches-Montagnes stallions. Methods: All horses were examined and assigned a dental malocclusion score by a veterinary dentist and randomized into two groups: sham treatment (Group S) and performance dentistry including occlusal equilibration (Group D). The horses were ridden twice before and three times after treatment by a professional dressage rider (unaware of treatment allocation). The horses were assigned a rideability score using a 27-point scale. The malocclusion score was compared to the average of the first two rideability scores using Spearmans coefficient of rank. Change in rideability scores over time was assessed by repeated measures ANOVA. Statistical significance was set at P < 0.05. Results: There was no correlation between dental score and rideability score prior to treatment (rs = 0.06, P = 0.73). In addition, there were no differences in the rideability score between treatment groups or as an interaction of treatment group and time (P = 0.93, P = 0.83, respectively). Conclusions: In conclusion, we were unable to show that performance dentistry improved equine rideability assessed by rider scoring. The addition of more objective measurement tools and a longer assessment period may help to scientifically prove what is anecdotally believed.