Respiratory disease and sero-epidemiology of respiratory pathogens in the working horses of Ethiopia.

BACKGROUND: Pathogens are frequently implicated in equine respiratory disease. In Ethiopia, respiratory disease is a frequent cause for presentation at veterinary clinics and a priority concern for users of working horses. However, there is little existing literature on possible aetiologies. OBJECTIVES: Determine prevalence of respiratory signs and exposure to major respiratory pathogens through a serological survey. STUDY DESIGN: Cross-sectional. METHODS: Systematically selected horses from 19 sites in central Ethiopia were examined clinically and sampled once (August-December 2013). A face-to-face interview collected data on horses' management and history. Serological testing targeted equine influenza virus (EIV), equine herpesviruses-1 (EHV-1) and -4 (EHV-4), equine rhinitis viruses A (ERAV) and B (ERBV), equine arteritis virus (EAV) and Streptococcus equi subspecies equi (S. equi). RESULTS: Owners reported a recent history of coughing in 38% of horses and nasal discharge in 7%. No animals were observed coughing during examination but 6% had a nasal discharge. Antibodies towards S. equi, were most prevalent (8%, 33/350). Antibodies to EAV were confirmed in one animal (0.3%). Low antibody titres to EHV-1/4 and ERA/BV suggested prior exposure but antibodies to EIV were not detected. Multivariable, multilevel logistic regression analysis for risk factors associated with S. equi serostatus showed higher odds of seropositivity in younger animals and those working less frequently. MAIN LIMITATIONS: A single serological sample cannot describe dynamic changes in antibodies. Sampling horses at the place of work may result in healthy-worker bias. CONCLUSIONS: S. equi may be endemic in this population and contributing, in part, to the occurrence of respiratory disease. Low prevalence of antibodies to viruses, with the exception of EIV, indicates these pathogens are present, but unlikely a predominant cause of respiratory signs and noninfectious causes of disease should also be investigated. Working horses in this region would be vulnerable to incursion of equine influenza.

Development and Evaluation of a Molecular Diagnostic Method for Rapid Detection of Histoplasma capsulatum var. farciminosum, the Causative Agent of Epizootic Lymphangitis, in Equine Clinical Samples.

Histoplasma capsulatum var. farciminosum, the causative agent of epizootic lymphangitis (EZL), is endemic in parts of Africa. Diagnosis based on clinical signs and microscopy lacks specificity and is a barrier to further understanding this neglected disease. Here, a nested PCR method targeting the internal transcribed spacer (ITS) region of the rRNA operon was validated for application to equine clinical samples. Twenty-nine horses with signs of EZL from different climatic regions of Ethiopia were clinically examined. Blood samples and aspirates of pus from cutaneous nodules were taken, along with blood from a further 20 horses with no cutaneous EZL lesions. Among the 29 horses with suspected cases of EZL, H. capsulatum var. farciminosum was confirmed by extraction of DNA from pus and blood samples from 25 and 17 horses, respectively. Positive PCR results were also obtained with heat-inactivated pus (24 horses) and blood (23 horses) spotted onto Whatman FTA cards. Two positive results were obtained among blood samples from 20 horses that did not exhibit clinical signs of EZL. These are the first reports of the direct detection of H. capsulatum var. farciminosum in equine blood and at high frequency among horses exhibiting cutaneous lesions. The nested PCR outperformed conventional microscopic diagnosis, as characteristic yeast cells could be observed only in 14 pus samples. The presence of H. capsulatum var. farciminosum DNA was confirmed by sequencing the cloned PCR products, and while alignment of the ITS amplicons showed very little sequence variation, there was preliminary single nucleotide polymorphism-based evidence for the existence of two subgroups of H. capsulatum var. farciminosum This molecular diagnostic method now permits investigation of the epidemiology of EZL.