The Burnout Syndrome (BOS) is a psycho-emotional disorder generating sustained stress over time, and inability to cope with the demands in an adaptive manner. The aims of the study were (1) to describe the prevalence of BOS during the third COVID-19 pandemic wave (May-June 2021); (2) to explore the relation of BOS with physical symptoms, and (3) to determine the profile favoring the development of BOS. This cross-sectional descriptive study was conducted amongst a sample of 759 healthcare professionals (HCPs). Data on sociodemographic variables, physical symptoms, the Maslach Burnout Inventory, Cervical Disability Index, and Numeric Pain Rating Scale were collected. The prevalence of BOS was 58.9% (447 subjects); 382 subjects (50.3%) showed high levels of Emotional Exhaustion (EE), whereas 219 participants (28.9%) had high Depersonalization (DP) levels, whilst 135 individuals (17.8%) showed low levels of Personal Fulfillment (PF). Women showed higher levels of EE (Z=-3.46; p = 0.001), whilst men showed higher levels of DP (Z=-2.69; p = 0.007). A total of 579 participants (76.3%) experienced muscle pain. A young nurse working in a hospital, or an emergency department emerges as a specific vulnerable profile. The current study confirms the need to overcome BOS in HCPs, implementing multidimensional tailored intervention to decrease the symptoms.