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Frontiers in veterinary science2025; 12; 1574351; doi: 10.3389/fvets.2025.1574351

Physiological and behavioral parameters of pain and stress in mares during and after transvaginal ultrasound-guided follicular aspiration.

Abstract: The use of transvaginal ultrasound-guided follicle aspiration (TVA) for oocyte collection has become a widely used procedure in horses for embryo production (IVEP). The TVA procedure is characterized by various manipulations, which are physical restraint, perineal preparation, transrectal palpation, insertion of an intravaginal device, and needle punctures. These repeated transvaginal and transovarian punctures have raised concerns about their potential effects on mare welfare. Our study aimed to investigate the effects of TVA manipulations, and especially puncturing, on pain and stress in mares in a commercial set-up. Unassigned: Therefore, eight mares were subjected to three TVA procedures: two with puncturing (P) and one without (P, control). Before, during and after all TVA procedures, blood was collected for serum glucose, lactate and cortisol levels, and facial pain scores were assessed. An electrocardiogram (ECG) was recorded for each mare before and during the procedure to measure heart rate (HR) and heart rate variability (HRV). Parameters in response to manipulations and puncturing were analyzed using linear mixed effect models for each outcome variable at different timepoints during the procedure. Unassigned: Results revealed that puncturing during TVA did not significantly influence serum metabolite levels, facial pain scores, HR, or HRV over the complete procedure ( > 0.05). Notably, HR did not increase at the moment of puncturing ( > 0.05), and no significant changes in HRV parameters between P and P were detected ( > 0.05). Both P and P procedures triggered significant increases in cortisol and lactate levels, facial pain scores, and HR during restraint in stocks and perineal preparation compared to the day before and the day after TVA ( < 0.05). Interestingly, even without puncturing, manipulations in the P procedure were sufficient to induce significant elevations in metabolite levels and facial pain scores compared to the day before and after ( < 0.05). Unassigned: These results indicate that the TVA procedure induces stress responses in mares, predominantly associated with all manipulations specific to TVA, while the effect of puncturing itself was minimal. All effects were acute, with parameters returning to baseline when measured 24 h later.
Copyright © 2025 Van den Branden, Salamone, Broothaers, Peere, Polfliet, Dewulf, Van Steenkiste, van Loon, Smits and Govaere.
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Publication Date: 2025-04-09 PubMed ID: 40271484PubMed Central: PMC12014748DOI: 10.3389/fvets.2025.1574351Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study investigates the effects of a common horse fertility procedure, transvaginal ultrasound-guided follicle aspiration (TVA), on mare welfare. It finds that while the procedure does cause some short-term stress, it doesn’t induce significant pain or long-term stress, regardless of whether the follicle aspiration is performed or not.

Objectives and Methodology

  • The primary goal of the research was to analyze the impact of TVA, specifically the needle punctures involved in the procedure, on the pain and stress levels in horses. The secondary purpose was to understand whether other aspects of the procedure cause stress or pain.
  • The study was carried out on eight mares. Each mare was subjected to three TVA procedures, two with puncturing and one without (which served as a control). All stages in the procedure were scrutinized, including physical restraint, perineal preparation, transrectal palpation, and the insertion of the intravaginal device.
  • Several parameters were measured before, during, and after the procedure, such as serum glucose, lactate and cortisol levels, facial pain scores, and heart-related parameters (HR and HRV) via an electrocardiogram. These varying parameters were analyzed using linear mixed effect models at different intervals during the procedure.

Results and Findings

  • The results showed that puncturing during TVA did not significantly affect serum metabolites, facial pain scores, heart rate, or heart rate variability.
  • Both TVA procedures with and without puncturing led to a momentary increase in cortisol and lactate levels, facial pain scores, and heart rate during the restraint and preparation stages compared to the levels observed the day before and after the procedure.
  • Surprisingly, even without puncturing, the TVA procedure’s manipulations led to a significant rise in metabolite levels and facial pain scores compared to the levels seen before and the day after the procedure.
  • It was underscored that the effects were acute and short-term, with all parameters returning to their initial state 24 hours post-procedure.

Conclusions

  • The study concluded that TVA does induce some short-term stress in mares, but these effects result mainly from the manipulations associated with the procedure, not the puncturing. Therefore, the welfare impact of TVA, as performed in this commercial setting, seems minimal.
  • It was highlighted that all the observed effects were temporary, suggesting that mares recover from the procedure within 24 hours.
  • This research can prove valuable for improving existing protocol for TVA and potentially mitigating the stress caused by the procedure.

Cite This Article

APA
Van den Branden E, Salamone M, Broothaers K, Peere S, Polfliet E, Dewulf M, Van Steenkiste G, van Loon G, Smits K, Govaere J. (2025). Physiological and behavioral parameters of pain and stress in mares during and after transvaginal ultrasound-guided follicular aspiration. Front Vet Sci, 12, 1574351. https://doi.org/10.3389/fvets.2025.1574351

Publication

Frontiers in veterinary science
ISSN: 2297-1769
NlmUniqueID: 101666658
Country: Switzerland
Language: English
Volume: 12
Pages: 1574351
PII: 1574351

Researcher Affiliations

Van den Branden, Emma
  • Department of Internal Medicine, Reproduction, and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Salamone, Matthieu
  • Department of Internal Medicine, Reproduction, and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
  • Division of Animal and Human Health Engineering, Department of Biosystems, KU Leuven, Leuven, Belgium.
Broothaers, Klaartje
  • Department of Internal Medicine, Reproduction, and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Peere, Sofie
  • Department of Internal Medicine, Reproduction, and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Polfliet, Ellen
  • Department of Internal Medicine, Reproduction, and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Dewulf, Manon
  • Department of Internal Medicine, Reproduction, and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Van Steenkiste, Glenn
  • Equine Cardioteam Ghent, Department of Internal Medicine, Reproduction, and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
van Loon, Gunther
  • Equine Cardioteam Ghent, Department of Internal Medicine, Reproduction, and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Smits, Katrien
  • Department of Internal Medicine, Reproduction, and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Govaere, Jan
  • Department of Internal Medicine, Reproduction, and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.

Conflict of Interest Statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Citations

This article has been cited 1 times.
  1. Márquez-Moya A, Carreras-Vico N, Sala-Ayala L, Martínez-Boví R, Cuervo-Arango J. 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 2026 Feb;61(2):e70183.
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