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Wendy Ángela González-López

Institute for Research and Technology in Food and Agriculture

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Neil Duncan

Institute for Research and Technology in Food and Agriculture

Sandra Ramos-Júdez

Institute for Research and Technology in Food and Agriculture

Juan F. Asturiano

Universitat Politècnica de València

Leonor Ferrão

Universitat Politècnica de València

Ignacio Giménez

Instituto Valenciano de Investigaciones Agrarias

Thales de Souza França

Universidade Federal do Rio Grande do Sul

Malbelys Padilla Sánchez

Universitat Politècnica de València

J. Beirão

Nord University

Alicia Felip

Instituto de Acuicultura Torre de la Sal

Danilo Pedro Streit

Universidade Federal do Rio Grande do Sul

Cooperative Institutions

Consejo Superior de Investigaciones Científicas

Universitat Politècnica de València

Institute for Research and Technology in Food and Agriculture

Instituto de Acuicultura Torre de la Sal

Centre National de la Recherche Scientifique

University of Algarve

Universitat Autònoma de Barcelona

Universidade Estadual Paulista (Unesp)

Instituto Nacional de Ciência e Tecnologia de Ciência Animal

Universitat de Barcelona

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Low sperm to egg ratio required for successful in vitro fertilisation in a pair-spawning teleost, Senegalese sole (Solea senegalensis)

bioRxiv (Cold Spring Harbor Laboratory) (2020)

Sandra Ramos-Júdez Wendy Ángela González-López Jhons Huayanay Ostos Noemí Cota Mamani Carlos Marrero Alemán

Abstract Cultured Senegalese sole ( Solea senegalensis ) breeders fail to spawn fertilised eggs and this bottleneck could be solved with the implementation of large-scale in vitro fertilisation protocols. However, low production of poor-quality sperm has frustrated the development of in vitro fertilisation protocols. Cultured females were induced to ovulate with a 5 µg kg -1 single injection of gonadotropin releasing hormone agonist (GnRHa) and good quality eggs (82.6 ± 9.2% fertilisation) were stripped 41:57 ± 1:46 h after the injection. Sperm was collected from cultured males, diluted in modified Leibovitz and used fresh to fertilise the eggs. A non-linear regression, an exponential rise to a maximum (R = 0.93, P < 0.0001) described the number of motile spermatozoa required to fertilise a viable egg and 1617 motile spermatozoa were sufficient to fertilise 99 ± 12% (± 95% CI) of viable eggs. Similar, spermatozoa egg -1 ratios of 592 ± 611 motile spermatozoa egg -1 were used in large-scale in vitro fertilisations with 190,512 ± 38,471 eggs. The sperm from a single male (145 ± 50 µL or 8.0 ± 6.8 × 10 8 spermatozoa) was used to fertilise the eggs. The mean hatching rate of the large-scale in vitro fertilisations was 70 ± 14 % to provide 131,540 ± 34,448 larvae per fertilisation. When unfertilised eggs were stored at room temperature the percentage of viable eggs decreased gradually and indicated the sooner eggs were fertilised after stripping the higher the viability of the eggs. The collection of sperm directly into a syringe containing modified Leibovitz significantly increased the percentage of motile spermatozoa (33.4 ± 12.2 %) compared to dilution in modified Leibovitz immediately after collection (6.6 ± 4.9 %). Senegalese sole have a pair-spawning reproductive behaviour characterised by external gamete fertilisation in close proximity with no sperm competition. The low spermatozoa egg -1 ratio required for maximum fertilisation was consistent with this reproductive behaviour and strategy. The provision of a large-scale in vitro fertilisation protocol (200 µL of sperm per 100 mL of eggs) will enable the industry to operate sustainably and implement breeding programs to improve production.

Fertilisation

Spawn (biology)

Milt

10.1101/2020.08.26.267740

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Citations (0)

Post-thawing dilution prolongs fish sperm use in aquaculture mediterranean species

Cryobiology (2023)

Thales de Souza França Wendy Ángela González-López Malbelys Padilla Sánchez Leonor Ferrão Fátima Fernández-García

Dilution

10.1016/j.cryobiol.2023.104715

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Citations (1)

Reproductive behaviour and fertilized spawns in cultured Solea senegalensis broodstock co-housed with wild breeders during their juvenile stages

General and Comparative Endocrinology (2024)

Wendy Ángela González-López Sandra Ramos-Júdez Neil Duncan

Broodstock

10.1016/j.ygcen.2024.114546

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Citations (4)

Automated Prediction of Spawning Nights Using Machine Learning Analysis of Flatfish Behaviour

Abdul Qadir Neil Duncan Wendy Ángela González-López Elvira Fatsini Francesc Serratosa

Senegalese sole (Solea senegalensis) broodstock exhibit distinct behaviours (Rest the Head, Guardian, Follow, and Locomotor activities) that are important for breeding success. Understanding and monitoring these behaviours are essential to understand successful breeding of Senegalese sole. However, manually analysing these behaviours represents a significant challenge for human observers and is a labour-intensive process. Therefore, to address these limitations, this study introduces a custom-designed framework based on computer vision and machine learning techniques. The model integrates object detection and tracking mechanisms to recognize and monitor reproductive behaviours of Senegalese sole within aquaculture environments. By combining convolutional neural networks (CNNs) with advanced tracking algorithms, our model effectively extracts and analyses behavioural patterns from video datasets. The results were also compared with manual analyses to identify key reproductive behaviours. The automated model demonstrated strong performance, with accuracy, precision, and specificity exceeding 87%, and a Pearson correlation of R = 0.99 between manual observation data and automated data. The model analysed videos to accurately identify behaviours with minimal human intervention, thereby saving a substantial number of hours and opened up the possibility to analyse behaviours over longer periods, generating more data. For the first time we analysed behaviours during the entire night and demonstrated how specific behaviours increased and decrease in relation to spawning. These changes in behaviour in relation to spawning enable the model to effectively correlate these behaviours with spawning and non-spawning nights to provide a predictive tool that identifies spawning with accuracies ranging from 70% to 100%.In conclusion, this study advances the capability of automated techniques in accurately analysing reproductive behaviour and predict spawning events in Senegalese sole. The model facilitate reproductive behavioural analyses and enable informed decisions for enhancing breeding success and promoting sustainable aquaculture practices.

Broodstock

Ethogram

10.2139/ssrn.5087039

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Successful cryopreservation in biodegradable containers of sperm from aquaculture Mediterranean fishes

Theriogenology (2023)

Thales de Souza França Wendy Ángela González-López Marivel Sánchez Leonor Ferrão F. Fernández-García

Cryoprotectant

Sea bass

10.1016/j.theriogenology.2023.12.016

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Citations (4)

Biodegradable capsules as an alternative container to sperm cryopreservation of aquaculture mediterranean fishes

Cryobiology (2023)

Thales de Souza França Wendy Ángela González-López Malbelys Padilla Sánchez Leonor Ferrão Álvaro Belenguer

10.1016/j.cryobiol.2023.104641

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Lipid and fatty acid composition of muscle, liver, ovary, and peritoneal fat in wild flathead grey mullet (Mugil cephalus) according to ovarian development

Theriogenology (2022)

Sandra Ramos-Júdez Alicia Estévez Wendy Ángela González-López Neil Duncan

Mugil

Flathead

Mullet

10.1016/j.theriogenology.2022.12.046

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Citations (8)

Sperm contamination by urine in Senegalese sole (Solea senegalensis) and the use of extender solutions for short-term chilled storage

Aquaculture (2019)

Wendy Ángela González-López Sandra Ramos-Júdez Ignacio Giménez Neil Duncan

Sperm Quality

10.1016/j.aquaculture.2019.734649

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Citations (16)

Providing recombinant gonadotropin-based therapies that induce complete oogenesis and produce viable larvae from an immature teleost, flathead grey mullet (Mugil cephalus)

bioRxiv (Cold Spring Harbor Laboratory) (2020)

Sandra Ramos-Júdez François Chauvigné Wendy Ángela González-López Hanna Rosenfeld Joan Cerdà

Abstract Under intensive captive conditions, wild-caught flathead grey mullet (Mugil cephalus) females remained arrested at early gonad development and no sperm could be obtained from males. With the aim to induce and complete oogenesis, induce the release of sperm and obtain fertilized eggs, adult female and male grey mullet were treated with M. cephalus single-chain recombinant gonadotropins (rGths), follicle-stimulating (rFsh) and luteinizing (rLh) hormones. In Experiment 1, fish were treated with a weekly dose of rFsh (15 μg kg−1), which in females significantly (P < 0.001) increased plasma concentration of 17β-estradiol and induced vitellogenic oocyte growth up to a maximum mean diameter of 425 ± 19 μm after 9 weeks of treatment. In Experiment 2, fish were treated with weekly injections of both rFsh and rLh at different doses (from 2.5 to 12 μg kg−1). Oocyte diameter reached 609 ± 5 μm, from which final oocyte maturation and ovulation was induced with 30 μg kg−1 of rLh and 40 mg kg−1 of progesterone. Good quality sperm (> 75% motile spermatozoa) was obtained from males in both experiments, and in Exp. 2 the addition of rLh induced the production of higher quantities of sperm that were used to fertilise the eggs. Although fertilisation was low (0.4 %), these fertilized eggs with embryo development produced viable larvae (71% hatching rate). This is the first report in a teleost species, to obtain larvae from eggs that were from immature females induced through to maturation with rGths. In comparison, control females remained arrested as immature fish and control males did not produce sperm. The study demonstrated that both rGths are effective to induce the entire process of oogenesis in sexually immature female grey mullet and to obtain flowing sperm from males, adding more data to confirm the roles of the Gths in teleost gametogenesis. This advance provides the bases of a therapy for the use in the aquaculture of teleost of commercial interest or the conservation of endangered species.

Mugil

Mullet

Flathead

Milt

Yolk

10.1101/2020.06.04.132175

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Gonadotropin induction of spermiation in Senegalese sole: Effect of temperature and stripping time

Aquaculture (2021)

François Chauvigné Jessica Lleberia Carlos Vilafranca Diogo Rosado M. de L. P. Martins

Efferent ducts

Gonadotropin

10.1016/j.aquaculture.2021.737844

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Citations (1)