Species identification of archived fish bones collected from the Mediterranean Sea 100 years ago using molecular techniques

With the discovery that DNA can be successfully recovered from museum collections, a new source of genetic information has been provided to extend our comprehension of the evolutionary history of species. However, historical specimens are often mislabeled or report incorrect information of origin, thus accurate identification of specimens is essential. Due to the highly damaged nature of ancient DNA many pitfalls exist and particular precautions need to be considered in order to perform genetic analysis. In this study we analyze 208 historical remains of pelagic fishes collected in the beginning of the 20th century. Through the adaptation of existing protocols, usually applied to human remains, we manage to successfully retrieve valuable genetic material from almost all of the examined samples using a guanidine and silica column-based approach. The combined use of two mitochondrial markers cytochrome-oxidase-1(mtDNA COI) and Control Region (mtDNA CR), and the nuclear marker first internal transcriber space (ITS1) allowed us to identify the majority of the examined specimens using traditional PCR and Sanger sequencing techniques. The creation of primers capable of amplifying heavily degraded DNA have great potential for future uses, both in ancient and in modern investigation. The methodologies developed in this study can in fact be applied for other ancient fish specimens as well as cooked or canned samples.

Geomorfologia del giacimento di orso delle caverne nella grotta di Govjestica, Valle di Praca, Bosnia-Ercegovina

In 2011 the GSB/USB caving group of Bologna has discovered, in the southern fossil branches of Govjestica cave (Valle di Praça, Bosnia) a fossil deposit of vertebrates containing bones of Ursus spelaeus, Capra ibex, Cricetulus migratorius and Microtus. On the basis of the U/Th ages of the bones, teeth and carbonate flowstone covering the fossils (60 ka), datings carried out in the laboratories of U-Series at Bologna, and on the disposition of the bones, a past connection between Govjestica and the nearby Banja Stjena cave is hypothesised. The closure of this passage has occurred suddenly through a collapse that has forced the last cave bears awakened from their winter sleep to stay blocked in Govjestica, and die. The connecting passage has later been covered with calcite flowstones and is no longer visible. This hypothesis is sustained by the rather scarce number of skeletons of cave bears found in Govjestica (a dozen of skulls against the often large amounts of cave bears found in similar caves): Govjestica cave, and especially the Room of the Bones in its southern part, has been used by cave bears only for a couple of centuries before these parts became inaccessible. Furthermore, the entrance of Banja Stjena cave was probably located close to or at the level of the Praça river, that has excavated its thalweg for around 20 metres in the last 60 ka.