Comparative Evaluation of Follicular Flushing Frequency and Scraping Time During Ovum Pick-Up in Mares: Effects on Oocyte Recovery Rate and Technical Considerations.
Abstract: Transvaginal ultrasound-guided follicular aspiration or ovum pick-up (OPU) has become the standard technique for oocyte collection in mares for intracytoplasmic sperm injection (ICSI). Although repeated follicular flushes and wall scraping are commonly used to improve oocyte recovery rate (ORR), the relative contribution of each remains unclear. This study aimed to compare the effects of multiple flushes versus controlled scraping time on ORR in mares. A controlled trial was conducted in two phases: (1) an ex vivo phase using slaughterhouse ovaries (n = 32), and (2) an in vivo phase in clinically healthy mares (n = 9). Follicles were assigned to two groups: multiple flushes (MF, 10 flushes with intermittent scraping, lasting 18 s in total) or single flush (SF, 18 s of continuous scraping followed by 1 flush). A total of 489 follicles were aspirated ex vivo and 143 in vivo. Results showed no significant difference in ORR between MF and SF groups in either phase (ex vivo: 63.8% vs. 59.5%; in vivo: 48.3% vs. 44.3%; p > 0.05). However, MF used significantly more flushing medium per follicle in the ex vivo model (p < 0.05). No significant differences were found in aspiration time or incidence of clots and blockages in the aspiration lines. A significant correlation was observed between clot number and aspiration system blockage (r = 0.497, p = 0.036). Continuous scraping without repeated flushing reduces medium usage but may increase operator fatigue and the risk of clot formation and system obstructions. Therefore, optimising scraping duration could improve the efficiency of oocyte recovery procedures while simplifying the technique and reducing costs. Further research is required to refine scraping protocols, reduce operator fatigue, and prevent complications related to clot formation.
© 2026 The Author(s). Reproduction in Domestic Animals published by Wiley‐VCH GmbH.
Publication Date: 2026-02-12 PubMed ID: 41674388PubMed Central: PMC12895297DOI: 10.1111/rda.70183Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Comparative Study
Cite This Article
APA
Márquez-Moya A, Carreras-Vico N, Sala-Ayala L, Martínez-Boví R, Cuervo-Arango J.
(2026).
Comparative Evaluation of Follicular Flushing Frequency and Scraping Time During Ovum Pick-Up in Mares: Effects on Oocyte Recovery Rate and Technical Considerations.
Reprod Domest Anim, 61(2), e70183.
https://doi.org/10.1111/rda.70183 Publication
Researcher Affiliations
- Equine Fertility Group, Facultad de Veterinaria, Universidad Cardenal Herrera-CEU, CEU Universities, Alfara del Patriarca, Spain.
- Equine Fertility Group, Facultad de Veterinaria, Universidad Cardenal Herrera-CEU, CEU Universities, Alfara del Patriarca, Spain.
- Equine Fertility Group, Facultad de Veterinaria, Universidad Cardenal Herrera-CEU, CEU Universities, Alfara del Patriarca, Spain.
- Equine Fertility Group, Facultad de Veterinaria, Universidad Cardenal Herrera-CEU, CEU Universities, Alfara del Patriarca, Spain.
- Equine Fertility Group, Facultad de Veterinaria, Universidad Cardenal Herrera-CEU, CEU Universities, Alfara del Patriarca, Spain.
MeSH Terms
- Animals
- Female
- Horses
- Oocyte Retrieval / veterinary
- Oocyte Retrieval / methods
- Ovarian Follicle
- Oocytes
Grant Funding
- GIR24-12 / Universidad San Pablo - CEU
Conflict of Interest Statement
: While preparing this work, ChatGPT was used as an AI‐assisted tool to refine the language and clarity of our written text. Following its use, we thoroughly reviewed and edited the content to ensure accuracy and appropriateness, taking full responsibility for the final published version of the article. The authors declare no conflicts of interest.
References
This article includes 22 references
- Alm, H. , Torner H., Kanitz W., Becker F., and Hinrichs K.. 1997. “Comparison of Different Methods for the Recovery of Horse Oocytes.” Equine Veterinary Journal 29: 47–50. 10.1111/j.2042-3306.1997.tb05099.x.
- Breton, A. L. , and Lewis N.. 2024. “Equine ART and Antral Follicle Count: Can We Deepen Our Understanding to Improve Outcomes?” Reproduction in Domestic Animals 59, no. S2: e14625. 10.1111/rda.14625.
- Brück, I. , Greve T., and Hyttel P.. 1999. “Morphology of the Oocyte‐Follicular Connection in the Mare.” Anatomy and Embryology 199, no. 1: 21–28. 10.1007/s004290050205.
- Brück, I. , Raun K., Synnestvedt B., and Greve T.. 1992. “Follicle Aspiration in the Mare Using a Transvaginal Ultrasound‐Guided Technique.” Equine Veterinary Journal 24, no. 1: 58–59. 10.1111/j.2042-3306.1992.tb02780.x.
- Claes, A. , and Stout T. A. E.. 2022. “Success Rate in a Clinical Equine In Vitro Embryo Production Program.” Theriogenology 187: 215–218. 10.1016/j.theriogenology.2022.04.019.
- Colleoni, S. , Barbacini S., Necchi D., Duchi R., Lazzari G., and Galli C.. 2007. “Application of Ovum Pick‐Up, Intracytoplasmic Sperm Injection and Embryo Culture in Equine Practice.” In Proceedings of the 53rd Annual Convention of the American Association of Equine Practitioners, 554–559. American Association of Equine Practitioners.
- Cuervo‐Arango, J. , Claes A. N., and Stout T. A.. 2019a. “A Retrospective Comparison of the Efficiency of Different Assisted Reproductive Techniques in the Horse, Emphasising the Impact of Maternal Age.” Theriogenology 132: 36–44. 10.1016/j.theriogenology.2019.04.010.
- Cuervo‐Arango, J. , Claes A. N., and Stout T. A. E.. 2019b. “Mare and Stallion Effects on Blastocyst Production in a Commercial Equine Ovum Pick‐Up–Intracytoplasmic Sperm Injection Program.” Reproduction, Fertility and Development 31, no. 12: 1894–1903. 10.1071/rd19201.
- Cuervo‐Arango, J. , Necchi D., Clutton‐Brock A., Profaska M., Crabtree J., and Claes A.. 2025. “Transvaginal Follicle Aspiration in Mares: A Description of Different Techniques and Comparison of Results Across Different OPU Clinics.” Reproduction in Domestic Animals 60, no. 3: e70043. 10.1111/rda.70043.
- Cuervo‐Arango, J. , Sala‐Ayala L., Márquez‐Moya A., and Martínez‐Boví R.. 2025. “The Influence of Aspiration Pressure, Follicle Flushing Method and Needle Rotation During Single‐Operator OPU Technique on Oocyte Recovery and Embryo Production in the Mare.” Animals 15, no. 6: 832. 10.3390/ani15060832.
- Felix, M. R. , Turner R. M., Dobbie T., and Hinrichs K.. 2022. “Successful In Vitro Fertilisation in the Horse: Production of Blastocysts and Birth of Foals After Prolonged Sperm Incubation for Capacitation.” Biology of Reproduction 107, no. 6: 1551–1564. 10.1093/biolre/ioac172.
- Fonte, J. S. , Alonso M. A., Junior M. P. M., et al. 2024. “Successful Equine In Vitro Embryo Production by ICSI – Effect of Season, Mares' Age, Breed, and Phase of the Estrous Cycle on Embryo Production.” Theriogenology 223: 47–52. 10.1016/j.theriogenology.2024.04.007.
- Gérard, N. , and Robin E.. 2019. “Cellular and Molecular Mechanisms of the Preovulatory Follicle Differenciation and Ovulation: What Do We Know in the Mare Relative to Other Species.” Theriogenology 130: 163–176. 10.1016/j.theriogenology.2019.03.007.
- Hawley, L. R. , Enders A. C., and Hinrichs K.. 1995. “Comparison of Equine and Bovine Oocyte‐Cumulus Morphology Within the Ovarian Follicle.” Biology of Reproduction 52: 243–252. 10.1093/biolreprod/52.monograph_series1.243.
- Hinrichs, K. , Schnobrich M., Fernandes C. B., Fleury P., Barillari V., and Bruggeworth S.. 2025. “Complications Associated With Equine Transvaginal Ultrasound‐Guided Follicle Aspiration: Practitioner Survey and Field Results.” Journal of Equine Veterinary Science 146: 105363. 10.1016/j.jevs.2025.105363.
- Lazzari, G. , Colleoni S., Crotti G., et al. 2020. “Laboratory Production of Equine Embryos.” Journal of Equine Veterinary Science 89: 103097. 10.1016/j.jevs.2020.103097.
- Márquez‐Moya, A. , Sala‐Ayala L., Carreras‐Vico N., Martínez‐Boví R., and Cuervo‐Arango J.. 2025. “Factors Influencing Oocyte Recovery During Ultrasound‐Guided Follicle Aspiration in Mares: A Postmortem Study.” Theriogenology 235: 39–45. 10.1016/j.theriogenology.2024.12.032.
- Rodriguez, J. , Maserati M., Robilotta T., et al. 2021. “Recovery of Equine Oocytes in Ambulatory Practice and Potential Complications.” Journal of Equine Veterinary Science 98: 103324. 10.1016/j.jevs.2020.103324.
- Russell, W. M. S. , and Burch R. L.. 1959. The Principles of Humane Experimental Technique. Methuen.
- Sala‐Ayala, L. , Pytel A. T., Stychno K., and Cuervo‐Arango J.. 2024. “Use of Excised Ovaries for Oocyte Recovery by Ultrasound Guided Follicular Aspiration – Validation of an Experimental Model for Research Purposes in Live Mares Ovum Pick Up.” Theriogenology 228: 9–16. 10.1016/j.theriogenology.2024.07.023.
- Scarlet, D. , Schuler G., Malama E., et al. 2025. “Endocrine Profile and OPU‐ICSI Outcomes in Mares: A Comparative Study.” Reproduction & Fertility 6, no. 3: e250027. 10.1530/raf-25-0027.
- Van den Branden, E. , Salamone M., Broothaers K., et al. 2025. “Physiological and Behavioral Parameters of Pain and Stress in Mares During and After Transvaginal Ultrasound‐Guided Follicular Aspiration.” Frontiers in Veterinary Science 12: 1574351. 10.3389/fvets.2025.1574351.
Citations
This article has been cited 0 times.Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists