Pseudomonas spp. isolated in diary herds. A new protocol with PCR real time by using “DUAL FRET” probes for ovine mastitis

Pseudomonas has been identified as an animals pathogen in particular it is reported occasionally as cause of bovine/ovine mastitis. Infections by this opportunistic pathogen are the leading cause of morbidity and mortality in dairy subjects. These bacteria are ubiquitous in soil and water, and on surfaces in contact with soil or water. Pseudomonas spp. has very simple nutritional requirements, in fact it is often observed growing in distilled water, which is evidence of its minimal nutritional needs. In veterinary field Pseudomonas mammary infection (in particular P. aeruginosa) may be peracute and severe due to endotoxin production, or it may be subacute or chronic, it’s tendency to colonize surfaces in a biofilm form makes the cells impervious to therapeutic concentrations antibiotics and biocides. In this work, we describe a novel and fast molecular approach for detect the initial pathogen biofilm core in dairy farms due at P. aeruginosa, P. fluorescens and P. putida. This strategy is based on early detection of the subsequent rate: Psudomonas vs total bacteria in biofilm specimens. This procedure is based on particular bi-functional FRET oligonucleotide probes named DUAL-FRET.