Data from the registration of flora and fauna species that took place within the Kharkiv as well as from Donetsk, Zaporizhzhia, Cherkasy administrative regions of Ukraine and Autonomous Republic of Crimea are presented in this dataset. A detailed description of biotopes is given, for migratory and introduced species the relevant status is mentioned. The dataset contains information on the number and density of the local populations or the number of individuals that were observed. Information on the latest and first registrations of migratory birds, active individuals of amphibians and reptiles, as well as imago stages of insects within the Kharkiv region is provided. Where possible, information subspecies taxonomic status is given. For particular records information about certain color variations (morphs), which is relevant for the sand lizard (Lacerta agilis), some diurnal butterflies, birds (owls (Strigiformes), diurnal birds of prey (Falconiformes, Accipitriformes)) is given.