Abstract: Background/objectives: DNA analysis is the most reliable method for the identification of human skeletal remains, especially the ones found in mass disasters or recovered from mass graves. To this aim, DNA was extracted from bones and teeth allegedly belonging to 27 Italian soldiers executed during the SecondWorldWar and exhumed from a mass grave in Ossero (Croatia). Methods: A selection of 131 different bone samples (petrous bones, femurs, metacarpal, and metatarsal bones) and 16 molar teeth were used for DNA extraction. Autosomal and Y-chromosome STR profiles were determined using a conventional CE approach, while a panel of 76 microhaplotypes was investigated through MPS. Results: Overall, 24 different autosomal consensus male profiles and six (unexpected) female profiles were identified; the male profiles were then compared with 21 alleged living relatives of the missing soldiers belonging to 14 unrelated Italian families. The DVI module of the Familias software was used for computing the LRs and the posterior probabilities (PP). The combination of autosomal STRs and microhaplotypes led to the identification of six victims and to a very likely identification of another one, supported by Y-haplotype sharing between victim and relative. Three distant victim–relative relationships resulting in low LR values for the autosomal markers showed Y-STR haplotype-sharing patterns, thus suggesting very strong support for a paternal relationship. Conclusions: The results of this study confirmed the effectiveness of the genetic approach and highlighted the presence of more individuals than expected in the mass grave, among which six were female subjects.
