Non-invasive approaches to morphological and molecular identification of insects from museum, archaeo-funerary and forensic contexts

Species identification is the starting point for the formulation of any further ipothesis in insect sciences and they are particularly important in forensic entomology and funerary arcaheoentomology. Two main approaches are currently used for identifying insects, the morphological one, preferred by old-school taxonomists and the more recent molecular one, favoured by non-specialists. Both approaches have respectively some limitations: lack of morphological identification keys, especially for the immature stages and the destruction of the specimens during the DNA extraction, as examples. The development of easy-access identification keys and of non-invasive DNA extraction methods permitting the repeatability of the analyses have been the main target of this thesis. To achieve this aims morphological analysis and DNA based methods were integrated and used on samples belonging to different contexts –forensic, museum and archaeological –and different timeframe –0 to 1000 years old-. The attention was paid to adults and the poor investigate puparia of flies from each context and the different timeframes.

For the DNA analysis commercial kits and homemade digestion buffers were tested on modern and old specimens but applying operational procedure to reserve the morphology of the specimens.

More then 10 puparia belonging to species in the family Heleopmyzidae, Sphaeroceridae, Piophilidae, Muscidae and Calliphoridae have been described and illustrated. Forty new DNA sequences, mainly from poor investigated taxa, were generated and 37 deposited in GenBank despite some problems arised, mainly due to presence of PCR inibithors in and on the cuticle.

The results, reported in this work, will impact in the anlysis of forensic and archaeological material but as well in the study of museum material, that especialy for the species typusis particularly precius and can not be destroid or altered during the identification process.

In addition specific question as the separation and identification of Hydrotaea puparia belonging to different species, the identification of poorly investigated species, the systematic of Sarcophaga species in the subgenus Sarcophaga and the evaluation of pre-Columbian funerary cerimonies have been answered. In addition the pictorial material produced will help entomologists working with fragmented material especially from old cases or archaeological sites. Several questions are still open and they request further investigations to be done in the future.