Development of high throughput molecolar diagnostic tools to monitor sheep and potentially zoonotic infectious diseases on bulk milk

Ovine farming makes a considerable contribution to total milk production, particularly in Mediterranean regions, in Central and Eastern Europe, and in the Middle East. Diseases caused by viral and microbial pathogens are responsable for the economic damage in sheep production. The zoonotic diseases are transmitted from animals to humans by direct contact or by contaminated milk, water, or the environment. Milk is a suitable matrice to detect pahogens involved in these infections. In Sardinia, animal farming can be considered one of the most important productive sectors. The aim of this project is the detection of microorganisms that cause some common diseases in Sardinia. Bulk Tank Milk (BTM) analysis was used to monitor health status in herds. In this project we developed qPCR assay for detecting and quantifying DNA of Coxiella burnetii, Mycoplasma agalactiae (Mag), Maedi-visna virus (MVV) and Mycobacterium avium subspecies paratubercolosis (MAP) in BTM samples. The results showed the presence of these microorganisms in different farms in Sardinia. This project showed that qPCR is more sensitive than others diagnostic assay, suggesting that it may become the "gold standard" for the detection of several microorganisms from biological samples. qPCR presented high sensitivity, specificity and reproducibility and allowed to analyze a large number of samples in a short time. Furthermore, this molecular assay allows timely diagnosis of infections by detection of very low quantity of the microorganisms DNA. Early detection of pathogenic microorganisms is of primary importance to allow prophylactic measures aimed to contain diseases diffusion in animals, and to identify the presence of zoonotic agents that could represent a serious threat to human health.

Ovine farming makes a considerable contribution to total milk production, particularly in Mediterranean regions, in Central and Eastern Europe, and in the Middle East. Diseases caused by viral and microbial pathogens are responsable for the economic damage in sheep production. The zoonotic diseasease are transmitted from animals to humans by direct contact or by contaminated milk, water, or the environment. Milk is a suitable matrice to detect pahogens involved in these infections. In Sardinia, animal farming can be considered one of the most important productive sectors. The aim of this project is the detection of microorganisms that cause some common diseases in Sardinia. Bulk Tank Milk (BTM) analysis was used to monitor health status in herds. In this project we developed qPCR assay for detecting and quantifying DNA of Coxiella burnetii, Mycoplasma agalactiae (Mag), Maedi-visna virus (MVV) and Mycobacterium avium subspecies paratubercolosis (MAP) in BTM samples. The results showed the presence of these microorganisms in different farms in Sardinia. This project showed that qPCR is more sensitive than others diagnostic assay, suggesting that it may become the "gold standard" for the detection of several microorganisms from biological samples. qPCR presented high sensitivity, specificity and reproducibility and allowed to analyze a large number of samples in a short time. Furthermore, this molecular assay allows timely diagnosis of infections by detection of very low quantity of the microorganisms DNA. Early detection of pathogenic microorganisms is of primary importance to allow prophylactic measures aimed to contain diseases diffusion in animals, and to identify the presence of zoonotic agents that could represent a serious threat to human health.