Diagnosis of exercise-induced bronchoconstriction (EIB) requires the use of bronchial provocation tests (BPTs). We assessed exercise-induced respiratory symptoms (EIRS), EIB and asthma in athletes and evaluated the validity of BPTs in the diagnosis of EIB. Rhinitis and atopy were also assessed. Athletes with (n = 55) and without previous asthma diagnosis (n = 145) were tested by skin prick tests, lung function and eNO measurements. EIRS were recorded and EIB was assessed by methacholine (Mch), eucapnic voluntary hyperpnoea (EVH), mannitol and exercise test. EIRS were highly reported and history of asthma was common among athletes. A high prevalence of atopy (48.7%) and allergic rhinitis (30.5%) was found. Athletes with asthma had a higher response rate to Mch and to EVH, as compared with athletes without a previous asthma diagnosis (P = 0.012 and P = 0.017 respectively). Report of EIRS, rhinitis and atopy were not associated with a positive BPT response. Screening athletes for EIB using BPTs is suggested, irrespective of reported EIRS or a previous asthma diagnosis.