Radiofrequency electromagnetic radiation (RF-EMZ) can affect semen quality and fertility in humans. However, the results of RF-EMZ research on the reproductive system of different species of domestic animals, especially boars, are very scarce. Therefore, the aim of this study was to investigate the influence of electromagnetic radiofrequency radiation at 2500 MHz on in vitro exposed semen samples of breeding boars, ie on spermatozoa motility and on the share of spermatozoa subpopulations using principal component and cluster analysis according to morphometric parameters of head and tail. Ejaculation, ie collection of samples, was performed by the method of manual fixation of the penis from a total of 12 boars, and semen samples of each boar were then distributed in Petri dishes to the experimental and control groups. Boar semens were irradiated in a gigahertz transverse electromagnetic (GTEM) chamber at a frequency of 2500 MHz and an electric field strength of 10 V / m for two hours. After irradiation, semen smears were stained with the Spermac method for the purpose of clearer visualization of morphological parts of spermatozoa. Morphometric analysis was performed with the computer program SFORM. The difference between individual spermatozoa morphometric parameters as well as spermatozoa motility of the experimental and control groups were not statistically significant. However, reduced motility of the experimental group (74.7%) compared to the control group of spermatozoa (85.7%) was observed. Progressive spermatozoa motility was statistically significantly reduced (p<0.001) in the experimental group of spermatozoa (35.0%) compared to the control group (60.1%). The analysis of the principal component revealed 4 components (head length and width, head regularity and middle circumference), while cluster analysis revealed 2 spermatozoa subpopulations (S1 subpopulation) characterized by spermatozoa of smaller length and more regular head shape and smaller midpiece circumference and (S2 subpopulation) characterized by spermatozoa of longer and more elongated head shape and higher circumference of the spermatozoa midpiece. The experimental group had a higher share of S1 subpopulation of spermatozoa (51.83%) compared to the control group (48.7%), as well as a lower share of S2 subpopulation (47.54%) compared to the control group (52.47%). The effect of radiofrequency 50 radiation at a frequency of 2500 MHz can negatively affect spermatozoa motility as well as the success of fertilization.