Strategies to eradicate paratuberculosis from cattle herds are based on the effective detection and removal of infected animals and improvement of animal husbandry to prevent new infections. Bacteriologic culture of Mycobacterium avium subsp. paratuberculosis (Map) from fecal samples is the most definitive method to detect infectious animals. Also in the Dutch program for Map unsuspected herds, culture of fecal samples is a key element. A disadvantage of fecal culturing is that the method is time consuming, requiring up to 16 weeks of incubation, labour intensive and thus expensive. Besides contamination by other bacteria and fungi is another problem with fecal cultures of Map. Contamination rates may be as high as 30%. As a result of these contaminated cultures, sampling and culturing have to be repeated, which also means loss of time and more costs. This high contamination rate undermined the motivation of farmers to participate in the Dutch paratuberculosis program. Therefore a project was started with the aim to reduce the contamination rate to below 10 %.