To date, there is very little information about the effects of ageing and sport selection on the characteristics of former athletes. The aim of this study was to analyse anthropometric and physiological characteristics of 146 former elite volleyball players in Italy. The possible effect of an active or inactive lifestyle on ageing was assessed by comparing the biological profiles of subsamples of former athletes with and without regular sport activity. All the former athletes underwent measurements of standard anthropometry (height, sitting height and body mass; skinfold thicknesses; bi-acromial and bi-cristal breadths; upper limb length; upper arm, calf and chest girths), cardio-respiratory function (vital capacity, forced vital capacity, forced expiratory volume in 1s; systolic and diastolic blood pressures and heart rate), muscle strength (handgrip strength), and cognitive function (visual and auditory simple reaction times; Digit-Symbol subtest). Body composition parameters were estimated from anthropometric measurements. Data on lifestyle were collected by questionnaires. The results show that the former players differ from current players in several anthropometric and physiological traits. Selection pressure seems to be more responsible for these differences than ageing. This study underlines the particular performance of former players in cognitive functions. Moreover, it confirms that an active lifestyle has beneficial effects on the biological age profiles of former athletes.