In The Netherlands, experimental vaccination against paratuberculosis with a killed vaccine proved to be successful in prevention of clinical paratuberculosis in the early sixties but was prohibited at that time due to cross-reactions with tuberculosis tests. This experiment was repeated in the eighties after The Netherlands were officially free of bovine tuberculosis with comparable results: a positive cost-benefit of vaccination due to a decrease in the rate of clinical paratuberculosis (Kalis CHJ et al.). Nowadays attention is not only focused on the reduction of the economic damage due to clinical paratuberculosis but also on the eradication of the disease from the herds. So, the question arises if vaccination can lead to a decrease or even elimination of Mycobacterium paratuberculosis (Mptb) in a herd. It has been suggested that freedom of infection might occur after 4 to 6 years of vaccination (Argente G.). The purposes of the present study were: (1) to study if vaccination against paratuberculosis during at least 10 years prevents faecal shedding of Mptb; (2) to compare the effect of a culture and cull eradication program on faecal shedding of Mptb between vaccinated and not vaccinated herds. The total number of faecal samples in ten herds with a history of more then ten years of vaccination was 750. Of these samples, 37 (4.9%) were Mptb positive. In three herds (30%) no Mptb positive faecal sample was cultured. In the seven positive herds the prevalence varied from 1% to 29 %. After a vaccination program of 12 years (5 of the 7 herds) the prevalence was 5.1%. In 39 herds with clinical paratuberculosis that never vaccinated against paratuberculosis a total number of 3045 faecal samples was cultured in the same period. Of these samples 182 were Mptb positive (6.0%). In seven herds (18%) no positive faecal sample was cultured. There was no significant difference between the vaccinated and the non-vaccinated herds in the rate of decline of culture-positives in the course of a two years culture and cull eradication program. In both groups the rate of faecal shedders declined over time with a sharp decline after the onset of the culture and cull program followed by a weak decline in the following period. This study demonstrates that a 10 years vaccination program is no guarantee that shedding of Mptb is prevented. Therefore a vaccination program has to be combined with, or replaced by a culture and cull program. The effect of a two year culture and cull program was not different between vaccinated and not vaccinated cows.