No evidence of bovine papillomavirus type 1 or 2 infection in healthy equids.

BACKGROUND: There is a large body of evidence supporting bovine papillomavirus types 1 and 2 (BPV1; BPV2) as aetiological agents of equine sarcoids. However, there is conflicting data regarding BPV1/2 infection in sarcoid-free equids. OBJECTIVES: Data obtained between 2007 and 2017 by BPV1/2 screening of sarcoids and nonsarcoid tumours vs. samples from healthy equids are presented to help clarify this issue. STUDY DESIGN: Cross-sectional study. METHODS: Tumour material obtained from horses, donkeys and mules with confirmed sarcoids (n = 130), suspected sarcoids (n = 120), or nonsarcoid lesions (n = 70), skin biopsies from 102 tumour-free horses and dandruff/hair roots from 35 tumour-free donkeys and mules were screened for BPV1/2 infection. Sample DNA was extracted and validated by equine ß-actin PCR. BPV1/2 screening was performed by BPV1/2 E5-specific PCR allowing for the detection of less than 10 viral DNA molecules. Twenty-six amplicons were bidirectionally sequenced and compared to known E5 variants using BLAST program. RESULTS: BPV1/2 E5 PCR scored positive for 130/130 diagnosed sarcoids, 63/120 suspected sarcoids and 13/70 nonsarcoid lesions, whereas 137/137 DNA aliquots derived from tumour-free equids tested negative. On predicted E5 protein level, six different BPV1 E5 variants were identified. MAIN LIMITATIONS: The diagnosis of equine sarcoid was not confirmed in 120 lesions. CONCLUSIONS: Lack of BPV1/2 E5 DNA in tumour-free equids and the prevalence of sarcoid disease in young adult individuals suggest that the time span between initial infection and sarcoid development is short. This contrasts with the long phase of virus latency characterising infection of humans by carcinogenic papillomaviruses. Presence of BPV1/2 DNA in several cases of poor wound healing/hypergranulation and dermatitis points to these skin disorders being possibly co-induced by BPV1/2. PCR screening of tumour tissue/scrapings for BPV1/2 DNA represents a reliable tool for the rapid validation of a clinical diagnosis of equine sarcoid.

The clinical diagnosis of equine sarcoids - Part 1: Assessment of sensitivity and specificity using a multicentre case-based online examination.

Equine clinicians and researchers often make the diagnosis of equine sarcoids (ES) on clinical assessment alone, without histopathological confirmation. However, the accuracy of the clinical diagnosis of ES has not been critically assessed. To assess sensitivity, specificity, positive and negative predictive values of the clinical diagnosis of ES, 40 clinical cases with histologically confirmed equine skin lesions (26 ES and 14 non-ES) were compiled in a representative online examination. For each case and lesion, at least one photograph and all relevant information were presented in a standardised format. This included the horse's signalment, lesion localisation, lesion progression, presence of other skin lesions, earlier treatments and response to treatment. No information relevant for the assessment of the lesion was intentionally withheld. Fourteen ES experts, 39 board-certified equine specialists, 103 equine practitioners and 25 novices categorised the cases into ES or non-ES and graded their level of diagnostic confidence on a scale from 1 to 6 for each case. The overall success rate was 82.0% while sensitivity and specificity were 83.3% and 79.6%, respectively. The positive and negative predictive values were 88.4% and 72.0%, respectively, in the tested population with a 66% prevalence of ES. However, less experienced veterinarians were frequently wrong in their clinical judgement despite a high level of diagnostic confidence. Therefore, the authors propose to develop a diagnostic tool to help clinicians optimise their selection of lesions requiring a biopsy. Ultimately, this will help reduce costs and morbidity generated by unnecessary diagnostic and therapeutic efforts.

Molecular allergen profiling in horses by microarray reveals Fag e 2 from buckwheat as a frequent sensitizer.

BACKGROUND: Companion animals are also affected by IgE-mediated allergies, but the eliciting molecules are largely unknown. We aimed at refining an allergen microarray to explore sensitization in horses and compare it to the human IgE reactivity profiles. METHODS: Custom-designed allergen microarray was produced on the basis of the ImmunoCAP ISAC technology containing 131 allergens. Sera from 51 horses derived from Europe or Japan were tested for specific IgE reactivity. The included horse patients were diagnosed for eczema due to insect bite hypersensitivity, chronic coughing, recurrent airway obstruction and urticaria or were clinically asymptomatic. RESULTS: Horses showed individual IgE-binding patterns irrespective of their health status, indicating sensitization. In contrast to European and Japanese human sensitization patterns, frequently recognized allergens were Aln g 1 from alder and Cyn d 1 from Bermuda grass, likely due to specific respiratory exposure around paddocks and near the ground. The most prevalent allergen for 72.5% of the tested horses (37/51) was the 2S-albumin Fag e 2 from buckwheat, which recently gained importance not only in human but also in horse diet. CONCLUSION: In line with the One Health concept, covering human health, animal health and environmental health, allergen microarrays provide novel information on the allergen sensitization patterns of the companion animals around us, which may form a basis for allergen-specific preventive and therapeutic concepts.

Safety and immunogenicity of BPV-1 L1 virus-like particles in a dose-escalation vaccination trial in horses.

REASONS FOR PERFORMING STUDY: Infection with bovine papillomaviruses types 1 and 2 (BPV-1, BPV-2) can lead to the development of therapy-resistant skin tumours termed sarcoids and possibly other skin diseases in equids. Although sarcoids seriously compromise the welfare of affected animals and cause considerable economic losses, no prophylactic vaccine is available to prevent this common disease. In several animal species and man, immunisation with papillomavirus-like particles (VLP) has been shown to protect efficiently from papillomaviral infection. HYPOTHESIS: BPV-1 L1 VLPs may constitute a safe and highly immunogenic vaccine candidate for protection of horses against BPV-1/-2-induced disease. METHODS: Three groups of 4 horses each received 50, 100 or 150 µg of BPV-1 L1 VLPs, respectively, on Days 0, 28 and 168. Three control horses received adjuvant only. Horses were monitored on a daily basis for one week after each immunisation and then in 2 week intervals. Sera were collected immediately before, 2 weeks after each vaccination and one and 2 years after the final boost and analysed by pseudovirion neutralisation assay. RESULTS: None of the horses showed adverse reactions upon vaccination apart from mild and transient swelling in 2 individuals. Irrespective of the VLP dose, all VLP-immunised horses had developed a BPV-1-neutralising antibody titre of ≥ 1600 plaque forming units (pfu)/ml 2 weeks after the third vaccination. Eight of 10 trial horses still available for follow-up had neutralising antibody titres ≥ 1600 pfu/ml one year and ≥ 800 pfu/ml 2 years after the last immunisation. CONCLUSION: Intramuscular BPV-1 L1 VLP vaccination in horses is safe and results in a long-lasting antibody response against BPV-1. Neutralisation titres were induced at levels that correlate with protection in experimental animals and man. POTENTIAL RELEVANCE: BPV-1 L1 VLPs constitute a promising vaccine candidate for prevention of BPV-1/-2-induced disease in equids.

Endoscopic electrosurgical excision of laryngeal granulation tissue in a standing pregnant mare.

An 11-year-old, eight-months pregnant mare suddenly developed severe respiratory distress. Endoscopic examination identified intralaryngeal granulation tissue obstructing the airway. In order to avoid general anaesthesia and the problems associated with a permanent tracheostomy, the mass was resected electrosurgically while the mare was standing. Patency was restored to the airway and the mare foaled without complication.