RESUMEN
The effects of radiofrequency electromagnetic radiation (RF-EMR) on semen quality have been in the spotlight in recent years, though research results to date have been contradictory. The effects of RF-EMR amongst others depend upon frequency, and there is currently no literature concerning the influence of 5G frequencies on both DNA integrity and spermatozoa vitality in males. The aim of this study was to investigate the effect of 5G RF-EMR on sperm membrane integrity, mitochondrial potential, and DNA integrity of in vitro exposed semen of breeding boars. The study included semen samples of eight breeding boars of the Pietren breed and four breeding boars of the German Landrace breed, from 1.5 to 3.5 years in age. Freshly diluted semen of each boar was divided into a control (n = 12) and experimental group (n = 12). The samples of the experimental group were exposed for 2 hours to continuous RF-EMR at a single frequency (700 MHz, 2500 MHz and 3500 MHz) and an electromagnetic field strength of 10 V/m using a transverse gigahertz electromagnetic cell. Sperm DNA fragmentation was assessed using a Halomax® kit and sperm membrane integrity and mitochondrial potential was assessed using a PI/SYBR-14 LIVE/DEAD viability kit with JC-1. A significantly higher proportion of spermatozoa with DNA fragmentation was found in exposed semen samples for all frequencies compared to the control group. The highest DNA damage was recorded in semen samples exposed to 5G RF-EMR at 2500 MHz (p < 0.01) and 3500 MHz (p < 0.05) vs. control semen samples. A significantly higher proportion of spermatozoa with damaged cell membrane and good mitochondrial potential was recorded in semen samples exposed with 3500 MHz. In vitro exposure of breading boar semen to 5G RF-EMR significantly increases the proportion of DNA fragmentation. The harmful effect of 5G RF-EMR on the proportion of spermatozoa with damaged DNA was frequency dependent. The 3500 MHz frequency displayed the most harmful effects due to significant impacts on DNA integrity and spermatozoa vitality indicators.