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Species identification of ancient Lithuanian fish remains using collagen fingerprinting

ABSTRACT: Morphological identification of ancient bone is often problematic due to heavy fragmentation that generally influences zooarchaeological assemblages. Fish bones are more taphonomically sensitive than those of other vertebrates as they are typically smaller and less biomineralised. Thus, taxonomic identification based on the preservation of morphological features is often extremely limited and can reduce or eliminate the usefulness of an assemblage for inferring taxon information. Currently, one of the most time- and cost-efficient methods of achieving faunal identity from ancient bone is by the collagen fingerprinting technique known as ZooMS (Zooarchaeology by Mass Spectrometry). ZooMS harnesses the potential of preserved collagen, which is the most dominant and time-stable protein in bone. In this research, ZooMS is applied to ancient Baltic region fish assemblages that are between 500 and 6000 years old in order to define species identity and construct assemblage compositions. Alongside inferences into environmental and biological shifts from the Neolithic era to present day in the Baltic region, we demonstrate for the first time the ability to distinguish between recently diverged members of the Salmo (salmon) and Scophthalmus (turbot) genera. ZooMS analysis highlights 7% of the collagen-containing assemblage as having been morphologically identified incorrectly and has facilitated taxonomic refinement of a further 28% of samples, including some of the morphologically indeterminate bone fragments. This research emphasises the great potential of ZooMS in identifying ichthyoarchaeological bone remains to species-level, and provides a case for the use of collagen fingerprinting in contributing to baseline fisheries and ecological data, to inform modern management.

INSTRUMENT(S): nanoACQUITY UPLC, LTQ Orbitrap Elite

ORGANISM(S): Salmo Trutta Fario Carassius Carassius Esox Lucius Silurus Glanis Scophthalmus Rhombus Sander Lucioperca Salmo Trutta Trutta Salmo Salar (atlantic Salmon) Gadus Morhua (atlantic Cod) Scophthalmus Maximus

TISSUE(S): Bone Tissue, Bone

SUBMITTER: Virginia Harvey

LAB HEAD: Michael Buckley

PROVIDER: PXD010041 | Pride | 2018-08-29

REPOSITORIES: Pride

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Project description:Groupers (Epinephelidae) are ecologically, commercially, and culturally important predatory fishes throughout their global distribution range in tropical, subtropical and occasionally temperate regions. They are key species for modern and ancient fisheries in the Mediterranean which have been heavily overfished in the past century leading to smaller catch sizes, lower CPUE, and decreased biomass. There are four species of grouper native to the Mediterranean within the Epinephelus genus.The abundance and distribution of grouper species prior to the 20th century in the Mediterranean remains poorly known. Using peptide mass fingerprinting, also known as Zooarchaeology by Mass Spectrometry (ZooMS), we investigated if ZooMS is a viable method for identifying intra-genus grouper bones to species level. Due to the lack of publicly available genomic sequences and for validation of ZooMS markers, we reconstructed collagen type I amino acid sequences using LC-MS/MS for four Epinephelus spp. Adequate variation between collagen sequences enabled the production of the best supported phylogenetic tree for Mediterranean Epinephelus spp. to date. We identified 23 previously undescribed ZooMS biomarkers capable of distinguishing groupers to the species level. Our novel biomarkers were applied to a case study of 23 grouper/comber fish bones from the Middle to Late Holocene archaeological site of Kinet Höyük, located along the coast of Iskenderun Bay, Turkey. ZooMS markers enabled species level identification of 19 bones with 18 identified as Epinephelus aeneus and 1 identified as Epinephelus marginatus. Combining ZooMS identifications with catch size reconstructions has revealed that E. aeneus is capable of growing ca. 30 cm larger than previously reported. This abundance and dominance of E. aeneus locally at Kinet Höyük is consistent with E. aeneus being the most prevalent grouper species in Iskenderun Bay today, testifying to several millennia of this species local population persistence despite fishing pressure, habitat degradation, and climatic changes.

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Species identification of ancient Lithuanian fish remains using collagen fingerprinting

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