Evaluation of methods to determine sperm density for the European eel, Anguilla anguilla

[EN] Contents European eel, Anguilla anguilla, is a target species for future captive breeding, yet best methodology to estimate sperm density for application in in vitro fertilization is not established. Thus, our objectives were to evaluate methods to estimate European eel sperm density including...

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Bibliographic Details
Published in:Reproduction in Domestic Animals
Main Authors: Sorensen, SR, Gallego Albiach, Victor, Pérez Igualada, Luz María, Butts, IAE, Tomkiewicz, J., Asturiano Nemesio, Juan Francisco
Other Authors: Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Universitat Politècnica de València, Ministerio de Ciencia e Innovación
Format: Article in Journal/Newspaper
Language:English
Published: Wiley 2013
Subjects:
Turbot scophthalmus-maximus
COD Gadus-morhua
Semen analysis
Hippoglossus-hippoglossus
Fertilization success
Spermatozoa density
Testis development
Gamete contact
Quality evaluation
Seminal plasma
PRODUCCION ANIMAL
Anguilla anguilla
European eel
Gadus morhua
Scophthalmus maximus
Turbot
Online Access:http://hdl.handle.net/10251/64788
https://doi.org/10.1111/rda.12189
Description
Summary:[EN] Contents European eel, Anguilla anguilla, is a target species for future captive breeding, yet best methodology to estimate sperm density for application in in vitro fertilization is not established. Thus, our objectives were to evaluate methods to estimate European eel sperm density including spermatocrit, computer-assisted sperm analysis (CASA) and flow cytometry (FCM), using Neubauer Improved haemocytometer as benchmark. Initially, relationships between spermatocrit, haemocytometer counts and sperm motility were analysed, as well as the effect of sperm dilution on haemocytometer counts. Furthermore, accuracy and precision of spermatocrit, applying a range of G-forces, were tested and the best G-force used in method comparisons. We found no effect of dilution on haemocytometer sperm density estimates, whereas motility associated positively with haemocytometer counts, but not with spermatocrit. Results from all techniques, spermatocrit, CASA and FCM, showed significant positive correlations with haemocytometer counts. The best correlation between spermatocrit and haemocytometer counts was obtained at 6000xg (r=0.68). Of two CASA variants, one or three photographic fields (CASA-1 and CASA-2), CASA-2 showed a very high accuracy to haemocytometer counts (r=0.93), but low precision (CV: CASA-2=28.4%). FCM was tested with and without microfluorospheres (FCM-1 and FCM-2), and relationships to haemocytometer counts were highly accurate (FCM-1: r=0.94; FCM-2: r=0.88) and precise (CV: FCM-1=2.5; FCM-2=2.7%). Overall, CASA-2 and FCM-1 feature reliable methods for quantification of European eel sperm, but FCM-1 has a clear advantage featuring highest precision and accuracy. Together, these results provide a useful basis for gamete management in fertilization protocols. This work was conducted as part of the project 'Reproduction of European Eel: towards a Self-sustained Aquaculture' (PRO-EEL) supported financially by the European Commission's 7th Framework Programme under the Theme 2 'Food, Agriculture and ...

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