Objective: The purpose of this retrospective study was to determine the vocal characteristics of a treatment-seeking population with the primary complaint of vocal fatigue (VF). Methods: Forty-three men (mean age 42 years, range 19-69) and 145 women (mean age 34 years, range 18-68) were included. None of the subjects had received voice therapy or previous laryngeal surgery. A questionnaire, laryngeal and perceptual evaluations, aerodynamic and acoustic parameters, and the Dysphonia Severity Index (DSI) were used to determine vocal characteristics. Results: In 74% of the subjects, flexible laryngeal videostroboscopic evaluation revealed a vocal pathology, with vocal nodules and muscle tension dysphonia as the most frequently diagnosed pathologies. Vocal abuse/misuse was present in 65% of the subjects. A median DSI value of -0.4 and -0.8 was found in female and male patients, respectively. Aerodynamic and acoustic parameters and DSI scores were significantly different from normative data. Conclusion: VF is a vocal sign with a significant need for medical consultation, especially in future professional voice users. Understanding the occurrence and the influencing variables of VF may help to close the gap between early stages of a vocal problem and the starting point of a well-established disorder.