ABSTRACT The present investigation aimed to evaluate the population structure and inbreeding of Holstein herds in southern Brazil. To carry out the analysis, the Associação Paranaense de Criadores de Bovinos da Raça Holandesa (APCBRH) in Brazil provided the data, which consisted of a pedigree file of 206,796 animals born between 1970 and 2014. Results regarding the following parameters were determined: pedigree integrity, effective number of founders, effective number of ancestors, generation interval, inbreeding coefficient, realized effective population size, and average relatedness coefficient. POPREP and ENDOG v.4.5 software packages were employed to estimate these parameters. Based on the data set, the mean generation interval was found to be 6.3 years, and the average inbreeding coefficient, related to inbred animals, was 4.99%. Furthermore, the realized effective population size varied throughout the time period, ranging from 22 to 114, whereas the rate of inbreeding in this same period showed a decreasing trend towards the later years in the period until 2014. Upon evaluation, average relatedness coefficient was estimated to be 0.71%. Moreover, the effective number of founders and ancestors were estimated as 418 and 400 animals, respectively. According to the level of inbreeding observed, it could be noticed that genetic diversity remains elevated, which will be important to the genetic progress in the Holstein breeding program in Southern Brazil.