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RESEARCH ARTICLE (Open Access)

Assessment of loliolide extracted from Biserula pelecinus, present during in vitro oocyte maturation, on fertilisation and embryo development in sheep

A. A. Amir A # , A. A. Algreiby B # , J. M. Kelly C , D. O. Kleemann https://orcid.org/0000-0002-4346-7730 C , Z. Durmic https://orcid.org/0000-0001-7300-043X A , G. R. Flematti D , D. Blache A and G. B. Martin https://orcid.org/0000-0002-1905-7934 A *
+ Author Affiliations
- Author Affiliations

A UWA School of Agriculture and Environment, and UWA Institute of Agriculture, The University of Western Australia, Crawley, WA 6009, Australia. Email: zoey.durmic@uwa.edu.au, dominique.blache@uwa.edu.au, annaamir79@gmail.com

B Department of Chemistry, College of Science, Qassim University, Buraydah, Saudi Arabia. Email: grieby@qu.edu.sa

C South Australian Research and Development Institute, Turretfield Research Centre, Rosedale, SA 5350, Australia. Email: jenkelly@internode.on.net, dave.kleemann@sa.gov.au

D School of Molecular Science, The University of Western Australia, Crawley, WA 6009, Australia. Email: gavin.flematti@uwa.edu.au

* Correspondence to: graeme.martin@uwa.edu.au

# These authors contributed equally to this paper

Handling Editor: Joanna Souza-Fabjan

Animal Production Science 64, AN24255 https://doi.org/10.1071/AN24255
Submitted: 3 August 2024  Accepted: 1 October 2024  Published: 29 October 2024

© 2024 The Author(s) (or their employer(s)). Published by CSIRO Publishing. This is an open access article distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND)

Abstract

Context

As a ‘duty of care’, it is important to test whether new forage plants for ruminants contain secondary compounds (PSCs) that affect reproductive performance. We have previously observed, a posteriori, that the presence of a methanolic extract of Biserrula pelecinus during maturation of sheep oocytes increased fertilisation rate and blastocyst development. This result needed to be verified a priori and, if the outcome was repeated, we needed to identify the plant secondary metabolite responsible.

Aims

To test whether PSCs from B. pelecinus, when added to the oocyte maturation medium, improve fertilisation rate and blastocyst development; to test whether loliolide is the active molecule produced by B. pelecinus.

Methods

Methanol–chloroform extracts of B. pelecinus were fractionated using rapid silica filtration and solvents of increasing polarity. Fractions at final concentrations of 0, 100 or 200 μg mL−1 were added to the medium used to mature sheep cumulus–oocyte complexes (COCs) and effects were determined for maturation, subsequent cleavage rate, blastocyst rate, hatching rate, blastocyst efficiency and total blastocyst cell number (TCN).

Results

Fraction BP-6 at 100 μg mL−1 reduced blastocyst rate (P < 0.05), but had no effect when the dose was doubled to 200 μg mL−1. Further fractionation using semi-preparative high-performance liquid chromatography showed loliolide as the most abundant compound in BP-6. Supplementation of the in vitro maturation medium with loliolide (0, 2.5, 5, 10 and 25 μg mL−1) did not affect any measure of embryo development. All COCs treated with B. pelecinus fractions reached the final stage of embryo development, blastocyst hatching. Total blastocyst cell number was not affected.

Conclusion

The presence of fractions of B. pelecinus extract during in vitro oocyte maturation can reduce embryo development.

Implications

In vitro techniques can detect potential effects of forages on reproduction. Some fractions from an extract of B. pelecinus when present during oocyte maturation can reduce embryo development. The abundant PSC, loliolide, was not responsible. There was no indication that a PSC in B. pelecinus improves outcomes.

Keywords: Biserrula pelecinus, blastocyst, fertilisation, IVEP, IVMFC, loliolide, oocyte, sheep.

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