Real-time PCR for detection of Brucella ovis and Histophilus somni in ovine urine and semen

Infectious ovine epididymitis affects rams worldwide, and it is considered to be one of the most important infectious diseases (Poester et al., 2013). Brucella ovis is one of the most important infectious agents associated with this disease, which may also be due to Histophilus somni infection, as well as other organisms (DíazAparicio et al., 2009; Poester et al., 2013; Moustacas et al., 2014). B. ovis infection is associated with chronic epididymitis and infertility in rams and, occasionally, abortion in ewes and birth of weak lambs. H. somni is a component of the microbiota colonizing mucosal surfaces of cattle, goats, and sheep. However, H. somni may act as an opportunistic pathogen, and the infection may result in epididymitis as well as several other clinical manifestations such as vaginitis,  placentitis, pneumonia, meningoencephalitis, mastitis, synovitis, septicemia, and other reproductive disorders (Díaz-Aparicio et al., 2009; Poester et al., 2013; Moustacas et al., 2014). The gold standard diagnosis of B. ovis infection is based on clinical examination, serology and semen bacteriology (Xavier et al, 2011; Poester et al., 2013), while for H. somni infections, only clinical examination and bacteriology are routinely performed. Semen and urine are the samples of choice for diagnosis (Xavier et al., 2010; Costa el al., 2012; Moustacas et al., 2013). These techniques are labor intensive and slow, whereas molecular techniques have been increasingly used, including conventional PCR (Xavier et al.,