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Barcoding and morphometry to identify and assess genetic population differentiation and size variability in loliginid squid paralarvae from NE Atlantic (Spain)

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Abstract

Accurate species classification is essential to understand complete life cycles of cephalopods. Identifying freshly caught or fixed loliginid paralarvae to species level with the traditional taxonomic guides is challenging. Therefore, the aim of this work was to identify genetically loliginid paralarvae captured in NW Spain (a region where at least three loliginid species are known to coexist) during 2012, 2013, and 2014, and to seek a means to distinguish the species from each other based on their morphometry. First, the barcoding region (COI gene) was amplified to identify each paralarva, and to obtain population molecular diversity indices and genetic structure for the different species. Afterwards, discriminant analysis (DA) was used to evaluate the performance of the selected morphometric measurements to distinguish among the species previously identified. Molecular analyses revealed three loliginid species (Alloteuthis media, A. subulata, and Loligo vulgaris), with different patterns of molecular diversity. DAs based on body morphometrics correctly categorised 75% of paralarvae to genus (Loligo and Alloteuthis) and 72% of Alloteuthis individuals to species level (A. media and A. subulata). When statolith measurements were included in the morphometric analysis, successful classification increased to 94 and 82%, respectively. The most useful variables for the discrimination of genus were hatching ring length and head width, while tentacle length helped to differentiate A. media from A. subulata. These discriminant functions should be tested with more paralarvae from different origins and seasons to account for body shape plasticity, but suggest a promising result to facilitate loliginid paralarvae identification for future research.

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Acknowledgements

We are indebted to the captain, crew, and technicians of R/V ‘Mytilus’ (IIM, CSIC Vigo) for their assistance in collecting the zooplankton samples. We are grateful to Mariana Cueto for assisting us with laboratory analyses and Lara García Alves for helping us to sort the paralarvae. This study was supported by the project LARECO (CTM-2011-25929) and CALECO (CTM2015-69519-R) funded by the Spanish Ministry of Economy and Competitiveness. Lorena Olmos-Pérez was supported with an FPI grant funded by the Spanish Ministry of Economy and Competitiveness (BES-2012-055651). We would like to thank the reviewers for their suggestions and comments that improved the quality of the manuscript.

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Correspondence to Lorena Olmos-Pérez.

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This study was performed in accordance with the existing Spanish guidelines and regulations on animal research (Ley 32/2007, November 7th), and was consequently exempt from an ethics review process.

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All the authors have revised the manuscript critically for important intellectual content and have approved the final version to be published. The authors declare that the research was conducted in the absence of any relationships that could be construed as a potential conflict of interest. We also declare that the manuscript has not been published previously or split up into several parts, neither the data have been fabricated or manipulated.

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Responsible Editor: R. Villanueva.

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Olmos-Pérez, L., Pierce, G.J., Roura, Á. et al. Barcoding and morphometry to identify and assess genetic population differentiation and size variability in loliginid squid paralarvae from NE Atlantic (Spain). Mar Biol 165, 136 (2018). https://doi.org/10.1007/s00227-018-3387-y

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  • DOI: https://doi.org/10.1007/s00227-018-3387-y

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