Introduction: Working tasks in the cleaning sector demand some high physical effort, affecting the heart rate variability (HRV) which is the measurement in the RR interval frequency between heartbeats. The purpose of this study was to describe the heart rate variability of non-athlete workers while performing a number of cleaning tasks. Material and methods: Observational cross-sectional pilot study. Working tasks from 20 cleaning workers were identified, followed by registering of their HRV by Holter monitor. Results: Mean age was 29.5 ± 6.56 years. Mean heart rate (HR) ranged from 85 to 92 bpm, remaining stable during cleaning tasks. High HR ranged from 119 to 225, showing negative asymmetry. Rotary floor machine cleaning, cleaning trolley pushing and wall washing produced higher mean RH, while window cleaning and vacuuming produced lower mean HR. Tasks like vacuuming and trolley pushing showed higher ranges in the RR interval, while toilet cleaning showed the lowest range. Conclusions: It was identified that vacuuming, rotary machine cleaning and cleaning trolley use produced higher heart rate variability. HRV is a parameter that requires further attention in workers’ health surveillance.