The genetic diversity of Coxiella burnetii from 36 dairy cattle herds was determined by Multiple-Locus Variable number tandem repeats Analysis (MLVA), and genotypes from different sources (bulk-tank milk – BTM and surface dust) and sampling time (2009/10 and 2011/12) were compared. A total of 15 different genotypes were identified from 60 BTM and seven dust samples, including seven genotypes reported here for the first time (BN, BO, BP, BQ, BR, BS, BT). The two most prevalent genotypes (J and I), detected both in BTM and dust, accounted for 44.5% of the C. burnetii typed and have been reported infecting cattle worldwide. In 52% of herds more than one genotype was found, and mixed infection with two genotypes was observed in seven BTM samples. Comparison of C. burnetii genotypes at different samplings within each herd detected a change in genotype in 32% of herds, while a persistent genotype was identified in the remaining 68%. In addition, the genotype obtained from dust samples was always identical to that present in the BTM sample. Often persistent genotypes were among the most prevalent types. Clustering of the MLVA genotypes from this and other studies using the minimum spanning tree method separated our C. burnetii strains into two clusters, 10 genotypes clustered within genomic group (GG) III, and the remaining five types (AE, BQ, BR, BS and BT) grouped with GG II, which includes strains implicated in human outbreaks. Although presence in cattle of genotypes closely related to those identified in humans does not seem to be common event, it cannot be neglected and surveillance of genotype distribution is needed to fully understand the epidemiology of Q fever.