Isolation of Mycobacterium avium Subsp. Paratuberculosis in the Feces and Tissue of Small Ruminants Using a Non-Automated Liquid Culture Method

摘要

In Italy, the field isolates of subsp. in bovine and goats (type II or C) are frequently found while the isolation of MAP in sheep (type I or S) is very rare. The aim of this study was to detect field isolates of MAP in ovine and goats using a non-automated method with liquid media culture. For this purpose, five flocks were investigated by ELISA and some feces of positive animals were tested by quantitative polymerase chain reaction (qPCR). All direct PCR positive samples were tested using a non-automated culture method using liquid media. Positive samples were analyzed by typing specific-PCR in order to determine the strain type (I or II). Twenty-eight samples, feces and tissues, were cultivated and 12 of these were positive by qPCR. From 12 positive samples, six strains resulted type I (S), from two flocks and four sheep respectively. We have evaluated a rapid and inexpensive culture method for the recovering of MAP type I, which are mycobacteria extremely growth demanding.