In situ sex identification in senegalese sole (Solea senegalensis) using loop-mediated isothermal amplification

The Senegalese sole (Solea senegalensis) is a promising candidate for aquaculture diversification in southern Europe due to its favorable traits and high market value. However, reproductive challenges and sexual dimorphism in growth necessitate efficient sex identification to optimize male-to-female...

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Main Authors: Emma Nicole Short, Laureana Rebordinos, Alejandro Centeno-Cuadros
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:Aquaculture Reports
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352513425003576
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author Emma Nicole Short
Laureana Rebordinos
Alejandro Centeno-Cuadros
author_facet Emma Nicole Short
Laureana Rebordinos
Alejandro Centeno-Cuadros
author_sort Emma Nicole Short
collection DOAJ
description The Senegalese sole (Solea senegalensis) is a promising candidate for aquaculture diversification in southern Europe due to its favorable traits and high market value. However, reproductive challenges and sexual dimorphism in growth necessitate efficient sex identification to optimize male-to-female ratios in farmed populations. Current sexing techniques are often invasive and time-consuming, prompting a need for non-invasive, reliable alternatives. To this end, this study presents the first loop-mediated isothermal amplification (LAMP) protocol for molecular sex identification in Senegalese sole. We designed novel primers targeting sex-linked SNPs in the fshr gene for both LAMP and PCR-based assays and extracted DNA from fresh tissue and non-invasively from skin mucus. An open-tube LAMP method was optimized for laboratory use, followed by a closed-tube LAMP method for in situ applications, incorporating the dye eriochrome black T to minimize contamination risks. Both LAMP approaches successfully identified male DNA with high specificity, demonstrating full concordance with PCR results. This innovative sexing technique for Senegalese sole enabled rapid, non-invasive sexing directly from mucus samples, making it suitable for applications outside of the modern laboratory environment, in both aquaculture facilities and natural environments.
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spelling doaj-art-cb5e45d722754f37a16d47adec5c7e6c2025-08-20T03:40:46ZengElsevierAquaculture Reports2352-51342025-09-014310297110.1016/j.aqrep.2025.102971In situ sex identification in senegalese sole (Solea senegalensis) using loop-mediated isothermal amplificationEmma Nicole Short0Laureana Rebordinos1Alejandro Centeno-Cuadros2Genetics Laboratory, Faculty of Marine and Environmental Sciences. University of Cádiz. C/ República Árabe Saharaui, s/n. 11510, Puerto Real, Cádiz, SpainGenetics Laboratory, Faculty of Marine and Environmental Sciences. University of Cádiz. C/ República Árabe Saharaui, s/n. 11510, Puerto Real, Cádiz, Spain; Instituto de Investigaciones Marinas (INMAR), Puerto Real, Cádiz, SpainGenetics Laboratory, Faculty of Marine and Environmental Sciences. University of Cádiz. C/ República Árabe Saharaui, s/n. 11510, Puerto Real, Cádiz, Spain; Instituto de Investigaciones Marinas (INMAR), Puerto Real, Cádiz, Spain; Corresponding author at: Genetics Laboratory, Faculty of Marine and Environmental Sciences. University of Cádiz. C/ República Árabe Saharaui, s/n. 11510, Puerto Real, Cádiz, Spain.The Senegalese sole (Solea senegalensis) is a promising candidate for aquaculture diversification in southern Europe due to its favorable traits and high market value. However, reproductive challenges and sexual dimorphism in growth necessitate efficient sex identification to optimize male-to-female ratios in farmed populations. Current sexing techniques are often invasive and time-consuming, prompting a need for non-invasive, reliable alternatives. To this end, this study presents the first loop-mediated isothermal amplification (LAMP) protocol for molecular sex identification in Senegalese sole. We designed novel primers targeting sex-linked SNPs in the fshr gene for both LAMP and PCR-based assays and extracted DNA from fresh tissue and non-invasively from skin mucus. An open-tube LAMP method was optimized for laboratory use, followed by a closed-tube LAMP method for in situ applications, incorporating the dye eriochrome black T to minimize contamination risks. Both LAMP approaches successfully identified male DNA with high specificity, demonstrating full concordance with PCR results. This innovative sexing technique for Senegalese sole enabled rapid, non-invasive sexing directly from mucus samples, making it suitable for applications outside of the modern laboratory environment, in both aquaculture facilities and natural environments.http://www.sciencedirect.com/science/article/pii/S2352513425003576LAMPfshrMolecular sexingEriochrome black T, non-invasive sampling
spellingShingle Emma Nicole Short
Laureana Rebordinos
Alejandro Centeno-Cuadros
In situ sex identification in senegalese sole (Solea senegalensis) using loop-mediated isothermal amplification
Aquaculture Reports
LAMP
fshr
Molecular sexing
Eriochrome black T, non-invasive sampling
title In situ sex identification in senegalese sole (Solea senegalensis) using loop-mediated isothermal amplification
title_full In situ sex identification in senegalese sole (Solea senegalensis) using loop-mediated isothermal amplification
title_fullStr In situ sex identification in senegalese sole (Solea senegalensis) using loop-mediated isothermal amplification
title_full_unstemmed In situ sex identification in senegalese sole (Solea senegalensis) using loop-mediated isothermal amplification
title_short In situ sex identification in senegalese sole (Solea senegalensis) using loop-mediated isothermal amplification
title_sort in situ sex identification in senegalese sole solea senegalensis using loop mediated isothermal amplification
topic LAMP
fshr
Molecular sexing
Eriochrome black T, non-invasive sampling
url http://www.sciencedirect.com/science/article/pii/S2352513425003576
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