Choir conductors represent a profession with an intensive vocal load. The present study aims to investigate the impact of 40 minutes vocal loading tasks on fundamental frequency (F0), jitter (Jitt), shimmer (Shim), median cepstral peak prominence (CPPs), and CPPs standard deviation in specific acoustic conditions. Method Eighteen amateur choir conductors (13 female, five male) participated in the study. The vocal loading experiment was organized in rooms where each conductor usually had choir rehearsals. The vocal loading experiment included vocal warm-up, singing, and loud reading. The STIPA test signal provided the background noise with 60 dBA @ 1m during the singing and reading tasks. The F0, Jitt, Shim, CPPs, and CPPs SD were extracted before and after vocal loading by the Vocal Holter Med (VHM) device. Additionally, Acoustic Voice Quality Index (AVQI) was estimated for participants, and all of them completed the Voice Handicap Index-30 (VHI-30) and the Singing Voice Handicap Index (SVHI). Finally, reverberation time (T30) was measured in each rehearsal room. Results A statistically significant decrease of CPPs SD by 0.1 dB was observed in male conductors. Associations between CPPs SD and VHI-30 (r = 0.789, P = 0.001) were found in female conductors, and between Jitt and SVHI (r = 0.9, P = 0.037) in male conductors. Female conductors with higher total score of SVHI demonstrated increase of postloading F0 (r = 0.612, P = 0.026). Conclusions Forty-minute vocal loading tasks that included warm-up, intensive singing, and loud reading did not significantly impact vocal parameters in female and male conductors. In addition, the rehearsal room volume and length of reverberation time did not affect obtained vocal parameters after vocal loading.