Serpentinites are metamorphic rocks with good technological properties and valuable ornamental characteristics, which have been exploited since ancient times. Actually, their use is limited and monitored in several countries worldwide because they can contain fibrous asbestos minerals that may be carcinogenic. Furthermore, certain types of fibrous minerals can be confused with asbestos, and must therefore be carefully investigated. We have investigated the possible presence of the asbestos and non-asbestos fibrous phases contained in serpentinitic rocks in a meta-ophiolitic sequence from the Gimigliano-Mount Reventino Unit (Southern Italy), which had not been previously assessed. The detection and quantification of asbestos and the correct distinction of the fibrous non-asbestos minerals are very important not only from a scientific point of view, but also from a legislative one. This is especially the case for the administrative agencies that have to take decisions with regards to the implementation of public and occupational health protection measures (e.g., in road yards and quarry excavations). As a consequence of this, serpentinitic rock samples have been characterized in detail through X-ray powder diffraction, scanning and transmission electron microscopy combined with energy-dispersive spectrometry, analytical electron microscopy (SEM–EDS and TEM–AEM), differential scanning calorimetry, thermogravimetry and micro-Raman spectroscopy. Two kinds of asbestos and four kinds of non-asbestos fibrous silicates have been detected in the examined samples. In order of decreasing abundance these are polygonal serpentine, chrysotile, fibrous antigorite, tremolite, gedrite and magnesiohornblende. The size, morphology, crystallinity and chemical composition of the fibres were also discussed, in the light of the possible role these properties could play in the carcinogenic effect on human health.