# Assessing past fish distribution through collagen fingerprints: a study on archaeological remains of Salmonids

Ambra d'Aurelio, Lucía Agudo Pérez, Camille Riquier, Pablo Arias, José Luis Arribas, Michel Barbaza, Eduardo Berganza, Manuel R. González Morales, Arturo Morales Muñiz, Jérôme Primault, et al.

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## To cite this version:

Ambra d'Aurelio, Lucía Agudo Pérez, Camille Riquier, Pablo Arias, José Luis Arribas, et al.. Assessing past fish distribution through collagen fingerprints: a study on archaeological remains of Salmonids. ADVANCING FISH ECOLOGY, MANAGEMENT AND FORECASTING THROUGH OMICS, Jul 2024, Bilbao, Spain. . [hal-05056791](https://hal.inrae.fr/hal-05056791v1)

HAL Id: hal-05056791

https://hal.inrae.fr/hal-05056791v1

Submitted on 5 May 2025

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# Assessing past fish distribution through collagen fingerprints: a study on archaeological remains of Salmonids

**Ambra D'Aurelio** (1, 2), Lucía Agudo Pérez (3), Camille Riquier (1, 2), Pablo Arias (4), José Luis Arribas (5), Michel Barbaza (6), Eduardo Berganza (7), Manuel R. González Morales (4), Arturo Morales-Muñiz (8), Jérôme Primault (9), Aurélien Simonet (10), Lawrence G. Straus (3, 11), Stephane Glise (1), Jean-Christophe Aymes (1), Frederic Lange (1), Emmanuel Huchet (1), Joëlle Chat* (1), Françoise Daverat* (1) and Ana B. Marín-Arroyo* (3) *Senior authors

**Affiliations:** (1) INRAE UMR 1224 ECOBIOP, 64310, Saint Pée sur Nivelle, France (2) Université de Pau et des Pays de l'Adour (UPPA), France (3) Grupo I+D+I EvoAdapta, Depto. de Ciencias Históricas, Universidad de Cantabria, Santander, Spain (4) Instituto Internacional de Investigaciones Prehistóricas de Cantabria, Universidad de Cantabria, Gobierno de Cantabria, Santander (5) AOZTA, Bilbao, Spain (6) TRACES UMR 5608, CNRS, UT2J Université de Toulouse Jean-Jaures, France (7) Sociedad de Ciencias Aranzadi, Donostia - San Sebastián, Spain (8) Laboratorio de Arqueozoología (LAZ-UAM)-Universidad Autónoma de Madrid, Darwin, 2, Madrid. Spain (9) DRAC/SRA Poitou-Charentes, Ministry of Culture and Communications, Poitiers, France (10) Service de la conservation des musées & du patrimoine, Direction de la culture et du patrimoine, Mont-de-Marsan, France (11) Department of Anthropology, MSC01 1040, University of New Mexico, Albuquerque, NM 87131-0001, USA

## Introduction

* The **Pleistocene-Holocene transition**, marked by dramatic climate changes during the Last Glacial Maximum (LGM, 26-19k BP) and the subsequent interglacial period<fnref n="1" />, played a crucial role in shaping modern biodiversity<fnref n="2" />.

* During the LGM, many species survived in **ice-free regions**, which are considered key areas for post-glacial recolonisation<fnref n="2" />. Nevertheless, just a few studies focused on the **past fish populations** that lived in these regions (e.g., <fnref n="3" />).

* Studying the **phenotypic diversity** of past species and their spatial-temporal dynamics is crucial for understanding adaptation strategies to climate change. The **brown trout** (*Salmo trutta* L.), known for its high intraspecific diversity, serves as an excellent model for such studies.

<figure><img src="image_3.png" /><figcaption>Fig 1. Life-cycle of brown trout (© Jean-Christophe Aymes)</figcaption></figure>

## Objective

This study aims to determine the **ecotype** and **size distribution** of brown trout during the Pleistocene-Holocene transition in the Iberian glacial refugium. The data can enhance our understanding of aquatic ecosystem responses to current climate change and inform effective management strategies.

## Material and Methods

We combined molecular and morphometric analyses on over **200 Upper Paleolithic vertebrae** (fig. 5) of brown trout from Cantabrian Spain and SW France (fig. 2).

<figure><img src="image_4.png" /><figcaption>Fig 2. Map of the 11 archaeological sites included in the study</figcaption></figure>

### 1) Analysis of collagen

* **ZooMS (Zooarchaeology by Mass Spectrometry):** species identification was performed by digesting extracted collagen type I with trypsin, producing species-specific peptides. These peptides were analyzed using a MALDI-TOF machine, returning the protein spectrum profile.

* **Stable isotope analysis (SIA):** The δ13C and δ15N were analysed. The δ13C was used to determine the ecotype (fig. 4), while δ15N helps study the trophic level and ecosystem structure.

### 2) Morphometric analysis

Three measurements (fig. 5) per vertebrae were taken using a calliper to reconstruct the size of the fish.

<figure><img src="image_5.png" /><figcaption>Fig 3. Collagen digestion through trypsin (adapted from <fnref n="4" />)</figcaption></figure>

<figure><img src="image_6.png" /><figcaption>Fig 4. Freshwater and seawater range of values for δ13C <fnref n="5" />, <fnref n="6" />, <fnref n="7" /></figcaption></figure>

<figure><img src="image_7.png" /><figcaption>Fig 5. Vertebra from Laminak II with the three measurements taken.</figcaption></figure>

## Results

### ZooMS for species identification

<figure><img src="image_8.png" /><figcaption>Reference peaks for *S. salar* and *S. trutta* from <fnref n="8" />, <fnref n="9" />, <fnref n="10" /></figcaption></figure>

### Size distribution of past brown trout

<figure><img src="image_9.png" /></figure>

### Life-history strategy through stable isotope of δ13C

<figure><img src="image_10.png" /></figure>

## Conclusions

1. ZooMS can successfully distinguish Paleolithic *S. trutta* and *S. salar* species.

2. Past brown trout size distribution: 30 – 60 cm

3. Both anadromous and resident ecotypes were present during the Pleistocene-Holocene transition

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<footnote n="1">¹ Clark, Peter U., et al. "The last glacial maximum." *science* 325.5941 (2009): 710-714.</footnote>

<footnote n="2">² Hewitt, Godfrey. "The genetic legacy of the Quaternary ice ages." *Nature* 405.6789 (2000): 907-913.</footnote>

<footnote n="3">³ Consuegra, S., et al. "Mitochondrial DNA variation in Pleistocene and modern Atlantic salmon from the Iberian glacial refugium." *Molecular ecology* 11.10 (2002): 2037-2048.</footnote>

<footnote n="4">⁴ Brown, Samantha, et al. "On the standardization of ZooMS nomenclature." *Journal of proteomics* 235 (2021): 104041.</footnote>

<footnote n="5">⁵ Almodóvar, Ana, et al. "Stable isotopes suggest the location of</footnote>

<footnote>marine feeding grounds of South European Atlantic salmon in Greenland," *ICES Journal of Marine Science* 77.2 (2020): 593-603.</footnote>

<footnote n="6">⁶ Cucherousset, Julien, et al. "Stable isotope evidence of trophic interactions between introduced brook trout *Salmo trutta* and native brown trout *Salmo trutta* in a mountain stream of south-west France." *Journal of Fish Biology* 71 (2007): 210-223.</footnote>

<footnote n="7">⁷ Musseau, C., et al. "Cannibalism in non-native brown trout *Salmo trutta* and rainbow trout *Oncorhynchus mykiss* stream-dwelling populations." *Journal of Fish Biology* 91.6 (2017): 1737-1744.</footnote>

<footnote n="8">⁸ Harvey, Virginia L., Linas Daugnora, and Michael Buckley. "Species identification of ancient Lithuanian fish remains using collagen fingerprinting." *Journal of Archaeological Science* 98 (2018): 102-111.</footnote>

<footnote n="9">⁹ Buckley, Michael, et al. "Medieval fish remains on the Newport ship identified by ZooMS collagen peptide mass fingerprinting." *Archaeological and Anthropological Sciences* 14.3 (2022): 41.</footnote>

<footnote n="10">¹⁰ Liz Quinlan, personal communication</footnote>