Evaluation of methods to determine sperm density for the european eel, anguilla anguilla
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,...
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ftdtupubl:oai:pure.atira.dk:publications/51205ca3-9eb8-4644-9a47-b6c1d33a456c 2023-10-01T03:50:17+02:00 Evaluation of methods to determine sperm density for the european eel, anguilla anguilla Sørensen, Sune Riis Gallego, V. Pérez, L. Butts, Ian Tomkiewicz, Jonna Asturiano, J. 2013 https://orbit.dtu.dk/en/publications/51205ca3-9eb8-4644-9a47-b6c1d33a456c https://doi.org/10.1111/rda.12189 eng eng info:eu-repo/semantics/closedAccess Sørensen , S R , Gallego , V , Pérez , L , Butts , I , Tomkiewicz , J & Asturiano , J 2013 , ' Evaluation of methods to determine sperm density for the european eel, anguilla anguilla ' , Reproduction in Domestic Animals , vol. 48 , pp. 936-944 . https://doi.org/10.1111/rda.12189 article 2013 ftdtupubl https://doi.org/10.1111/rda.12189 2023-09-06T22:59:00Z 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 6000 × g (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 Article in Journal/Newspaper Anguilla anguilla European eel Technical University of Denmark: DTU Orbit Reproduction in Domestic Animals 48 6 936 944 |
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Open Polar |
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Technical University of Denmark: DTU Orbit |
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ftdtupubl |
language |
English |
description |
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 6000 × g (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 |
format |
Article in Journal/Newspaper |
author |
Sørensen, Sune Riis Gallego, V. Pérez, L. Butts, Ian Tomkiewicz, Jonna Asturiano, J. |
spellingShingle |
Sørensen, Sune Riis Gallego, V. Pérez, L. Butts, Ian Tomkiewicz, Jonna Asturiano, J. Evaluation of methods to determine sperm density for the european eel, anguilla anguilla |
author_facet |
Sørensen, Sune Riis Gallego, V. Pérez, L. Butts, Ian Tomkiewicz, Jonna Asturiano, J. |
author_sort |
Sørensen, Sune Riis |
title |
Evaluation of methods to determine sperm density for the european eel, anguilla anguilla |
title_short |
Evaluation of methods to determine sperm density for the european eel, anguilla anguilla |
title_full |
Evaluation of methods to determine sperm density for the european eel, anguilla anguilla |
title_fullStr |
Evaluation of methods to determine sperm density for the european eel, anguilla anguilla |
title_full_unstemmed |
Evaluation of methods to determine sperm density for the european eel, anguilla anguilla |
title_sort |
evaluation of methods to determine sperm density for the european eel, anguilla anguilla |
publishDate |
2013 |
url |
https://orbit.dtu.dk/en/publications/51205ca3-9eb8-4644-9a47-b6c1d33a456c https://doi.org/10.1111/rda.12189 |
genre |
Anguilla anguilla European eel |
genre_facet |
Anguilla anguilla European eel |
op_source |
Sørensen , S R , Gallego , V , Pérez , L , Butts , I , Tomkiewicz , J & Asturiano , J 2013 , ' Evaluation of methods to determine sperm density for the european eel, anguilla anguilla ' , Reproduction in Domestic Animals , vol. 48 , pp. 936-944 . https://doi.org/10.1111/rda.12189 |
op_rights |
info:eu-repo/semantics/closedAccess |
op_doi |
https://doi.org/10.1111/rda.12189 |
container_title |
Reproduction in Domestic Animals |
container_volume |
48 |
container_issue |
6 |
container_start_page |
936 |
op_container_end_page |
944 |
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