Identification of Prototheca species responsible for bovine mastitis by new multiplex PCR method

Within the known Prototheca spp., P. zopfii genotype 2, P. wickerhamii and P. blaschkeae are algal pathogens associated with pathology in humans and animals. Different molecular techniques have been proposed to identify these pathogens, however such of these methods are unable to differentiate among different Prototheca species or are time consuming, demanding more than a PCR reaction. The purpose of the present study was to developed a multiplex PCR reaction, based on specific amplification of mithocondrial and chloroplastic sequences, able to identify in a single reaction the three Prototheca spp. causing bovine mastitis. Between the 30 strains collected from IMI milk samples and bulk tank milk, our preliminary results, obtained by this rapid and specific assay, showed an high prevalence of P. zopfii genotype 2 in Italian herd.