Abstract
BackgroundInfectious ovine epididymitis results in substantial economic losses worldwide due to reproductive failure and culling of breeders. The most common causative agents of these infections are Brucella ovis, Actinobacillus seminis, and Histophilus somni. The aim of this study was to develop a multiplex PCR assay for simultaneous detection of Brucella ovis, Actinobacillus seminis, and Histophilus somni with species-specific primers applied to biological samples for molecular diagnosis of these infections.ResultsThe multiplex assay was capable of detecting B. ovis, A. seminis, and H. somni DNA simultaneously from genomic bacterial DNA samples and pool of semen samples from experimentally infected rams. The method was highly specific since it did not amplify DNA from other bacterial species that can potentially cause epididymitis in rams as well as species phylogenetically related to B. ovis. All negative control samples were negative in PCR multiplex assay. Urine can be used as an alternative to semen samples.ConclusionsThe species-specific multiplex PCR assay developed in this study can be successfully used for the detection of three of the most common bacterial causes of ovine epididymitis.
Highlights
Infectious ovine epididymitis results in substantial economic losses worldwide due to reproductive failure and culling of breeders
A. seminis is a component of the normal flora of the prepucial mucosa, but it can act as opportunistic pathogen causing ascending infection which may lead to epididymitis and orchitis, in young rams [1,2]
Serologic tests are not widely available for diagnosis of A. seminis and H. somni infections and, the diagnosis is commonly based on clinical evaluation and semen bacteriology, PCR has been proposed as an alternative diagnostic method [10]
Summary
Infectious ovine epididymitis results in substantial economic losses worldwide due to reproductive failure and culling of breeders. The aim of this study was to develop a multiplex PCR assay for simultaneous detection of Brucella ovis, Actinobacillus seminis, and Histophilus somni with species-specific primers applied to biological samples for molecular diagnosis of these infections. Serologic tests are not widely available for diagnosis of A. seminis and H. somni infections and, the diagnosis is commonly based on clinical evaluation and semen bacteriology, PCR has been proposed as an alternative diagnostic method [10]. These infections are usually unresponsive to antibiotic treatment [11], resulting in considerable economic losses due to reproductive failure and culling of breeders [12]
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