Analyze Diet
Reproduction in domestic animals = Zuchthygiene2025; 60(10); e70131; doi: 10.1111/rda.70131

Prediction Values for the Influence of Fetal Sex on Plasma Progesterone Concentration in Crioulo Breed Mares: A Preliminary Investigation.

Abstract: In equine reproduction, determining foetal sex is of economic and strategic importance, but currently available methods are often invasive, costly, or require specialised expertise. This study aimed to assess whether plasma progesterone concentrations could serve as a predictive tool for foetal sex determination in Criollo mares between the 4th and 8th months of gestation and to determine whether these measurements could identify foetal sex. Blood samples were collected at 30-day intervals from 17 Crioula mares between 114 days and 240 days of gestation. Maternal plasma progesterone concentrations were determined by radioimmunoassay and the sex of the foals was confirmed at birth. Analysis of Variance (ANOVA) was made to verify the variation in maternal progesterone concentrations according to foetal sex and month of gestation. In mares carrying male foetuses (n = 7), there was a significant difference in progesterone concentrations between months 4 and 8, as well as between months 7 and 8. In mares carrying male foetuses (n = 7), progesterone concentrations were higher (p = 0.028) during the 4th month of gestation and lower at the 8th month (p = 0.020), compared to the values in mares with female foetuses (n = 10). In the 8th month of pregnancy, the sensitivity and specificity of progesterone concentrations for the predicted sex were 80% and 100%, respectively. A limit value of progesterone (12.45 ng/mL) was established through the ROC (receiver operating characteristic) Curve. Prediction values were 78.8% and 100% for males and females, respectively. Detection rates were 100% and 80% for males and females, respectively. The diagnostic accuracy for both sexes was 88.2%. It is concluded that foetal sex influences plasma concentrations of progesterone in pregnant mares.
Publication Date: 2025-10-07 PubMed ID: 41055121PubMed Central: PMC12502008DOI: 10.1111/rda.70131Google 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

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.

Overview

  • This study investigated whether measuring plasma progesterone levels in pregnant Criollo mares can help predict the sex of the foal between 4 and 8 months of gestation.
  • The research found significant differences in progesterone concentrations depending on fetal sex, suggesting progesterone as a potential non-invasive tool for fetal sex determination in horses.

Background and Purpose

  • In equine reproduction, knowing the sex of the fetus beforehand has both economic and strategic value for breeders and trainers.
  • Existing methods to determine fetal sex are often invasive, expensive, or require specialized practitioners.
  • The study focused on Criollo mares, a specific horse breed, aiming to explore if plasma progesterone levels—a hormone involved in pregnancy maintenance—can reflect fetal sex during mid-gestation (4 to 8 months).

Study Design and Methods

  • A total of 17 Criollo mares were included in the study, all between 114 and 240 days (roughly 4 to 8 months) pregnant.
  • Blood samples were collected every 30 days during this period to measure maternal plasma progesterone levels using radioimmunoassay, a sensitive hormone detection technique.
  • The foetal sex was confirmed at birth to compare with the progesterone data.
  • Statistical analysis using Analysis of Variance (ANOVA) was conducted to assess differences in progesterone levels by fetal sex and month of gestation.

Key Findings

  • In mares carrying male fetuses (n=7):
    • Progesterone levels were significantly higher during the 4th month of gestation compared to mares with female fetuses.
    • Progesterone levels dropped significantly by the 8th month, being lower than in females at this time.
    • Significant differences were noted between months 4 and 8, and months 7 and 8 for males.
  • In mares carrying female fetuses (n=10), progesterone levels exhibited different patterns, generally lower at month 4 and higher by month 8 compared to males.
  • At the 8th month of pregnancy, plasma progesterone concentration could predict fetal sex with:
    • 80% sensitivity for males (true positive rate)
    • 100% specificity for males (true negative rate)
  • A progesterone cutoff value of 12.45 ng/mL was identified using ROC curve analysis to differentiate male and female fetuses.
  • Prediction accuracy was high:
    • 78.8% predictive value for males
    • 100% predictive value for females
    • Detection rates of 100% (males) and 80% (females)
    • Overall diagnostic accuracy was 88.2%

Conclusions and Implications

  • The study concludes that fetal sex influences maternal plasma progesterone concentrations during gestation in Criollo mares.
  • Plasma progesterone has potential as a non-invasive, cost-effective biomarker to predict fetal sex in mares during mid to late pregnancy.
  • This method could offer advantages over traditional fetal sexing approaches that are invasive or require specialized skills.
  • Since this is a preliminary investigation with a modest sample size, further studies are recommended to confirm these findings and refine prediction protocols.

Cite This Article

APA
de Lara NSS, Weiss RR, Oba E, Kozicki LE, Souza FA, Bergstein-Galan TG, Muehlbauer E, Silvestri M, de Lima PHL, Vaz ES. (2025). Prediction Values for the Influence of Fetal Sex on Plasma Progesterone Concentration in Crioulo Breed Mares: A Preliminary Investigation. Reprod Domest Anim, 60(10), e70131. https://doi.org/10.1111/rda.70131

Publication

ISSN: 1439-0531
NlmUniqueID: 9015668
Country: Germany
Language: English
Volume: 60
Issue: 10
Pages: e70131
PII: e70131

Researcher Affiliations

de Lara, Natália Santana Siqueira
  • Universidade Federal Do Paraná - Curitiba, Paraná, Brazil.
Weiss, Romildo Romualdo
  • Universidade Federal Do Paraná - Curitiba, Paraná, Brazil.
Oba, Eunice
  • Universidade Estadual Julio Mesquita - Botucatu, São Paulo, Brazil.
Kozicki, Luiz Ernandes
  • Pontifícia Universidade Católica - Curitiba, Paraná, Brazil.
Souza, Fernando Andrade
  • Universidade Federal Do Paraná - Curitiba, Paraná, Brazil.
Bergstein-Galan, Tacia Gomes
  • Universidade Estadual de Ponta Grossa - Ponta Grossa, Paraná, Brazil.
Muehlbauer, Eloisa
  • Universidade Federal Do Paraná - Curitiba, Paraná, Brazil.
Silvestri, Mayara
  • Universidade Federal Do Paraná - Curitiba, Paraná, Brazil.
de Lima, Pedro Henrique Lomba
  • Pontifícia Universidade Católica - Curitiba, Paraná, Brazil.
Vaz, Eduarda Stankiwich
  • Universidade Federal Do Paraná - Curitiba, Paraná, Brazil.

MeSH Terms

  • Animals
  • Progesterone / blood
  • Horses / blood
  • Horses / physiology
  • Female
  • Pregnancy
  • Male
  • Sex Determination Analysis / veterinary
  • Sex Determination Analysis / methods
  • Sensitivity and Specificity
  • Gestational Age

Conflict of Interest Statement

The authors declare no conflicts of interest.

References

This article includes 34 references
  1. Aurich C, Schneider J. Sex Determination in Horses‐Current Status and Future Perspectives. Animal Reproduction Science 146, no. 1–2: 34–41.
  2. Barnes RJ, Comline RS, Silver M. Effect of Cortisol on Liver Glycogen Concentrations and in Hypophysectomised Adrenalectomized and Normal Foetal Lambs During Late or Prolonged Gestation. Journal of Physiology 275: 567–579.
  3. Bucca S. Equine Fetal Gender Determination From Mid‐ To Advanced Gestation by Ultrasound. Theriogenology 64, no. 3: 568–571.
  4. Busato EM, Weiss RR, Abreu ACMR. Correlation of Maternal Concentrations of Plasma Testosterone With Fetal Sex in Horses. Ciência Rural 51: e20200237.
  5. Caixeta ES, Fagundes NS, Caixeta MS, Pyles ES. Desenvolvimento Embrionário Inicial Equino – Revisão. Revista Portuguesa de Ciências Veterinárias 103, no. n.1: 25–34.
  6. Chezum B, Wimmer B. Roses or Lemons: Adverse Selection in the Market for Thoroughbred Yearlings. Review of Economics and Statistics 79, no. 3: 521–526.
  7. Clifton VL. Sex and the Human Placenta: Mediating Differential Strategies of Fetal Growth and Survival. Placenta 31, no. Suppl: S33–S39.
    pubmed: 20004469
  8. Curran S. Fetal Sex Determination in Cattle and Horses by Ultrasonography. Theriogenology 37, no. 1: 17–21.
  9. Curran S, Ginther OJ. Ultrasonic Diagnosis of Equine Fetal Sex by Location of the Genital Tubercle. Journal of Equine Veterinary Science 9, no. 2: 77–83.
  10. Curran S, Ginther OJ. Ultrasonic Fetal Gender Diagnoses During Months 5 to 11 in Mares. Theriogenology 40: 1127–1135.
  11. De Leon PMM, Campos VF, Dellagostin OA. Equine Fetal Sex Determination Using Circulating Cell‐Free Fetal DNA (ccffDNA). Theriogenology 77, no. 3: 694–698.
    doi: 10.1016/0378-4320(89)90095-Xpubmed: 22000028google scholar: lookup
  12. Dhakal P, Tsunoda N, Nakai R. Annual Changes in Day Length, Temperature, and Circulating Reproductive Hormones in Thoroughbred Stallions and Geldings. Journal of Equine Science 22, no. 2: 29–36.
    doi: 10.1294/jes.22.29pmc: PMC4013971pubmed: 24833985google scholar: lookup
  13. Duer C, Carden M, Tomasi T. Detection of Fetal Gender Differences in Maternal Serum Progesterone Concentrations of Asian Elephants (). Animal Reproduction Science 97, no. 1–2: 278–283.
  14. England, G. 2005. Fertility and Obstetrics in the Horse. Third ed. John Wiley & Sons.
  15. Holder RD. Fetal Sex Determination. Current Therapy in Equine Reproduction 12, no. 3: 8741.
  16. Holtan DW, Nett TM, Estergreen VL. Plasma Progestagens in Pregnant Mares. Journal of Reproduction and Fertility Supplement 23: 419–424.
    pubmed: 1060818
  17. Holtan DW, Nett TM, Estergreen VL. Plasma Progestagens in Pregnant Mares. Journal of Reproduction and Fertility Supplement 23: 419–424.
    pubmed: 1060818
  18. Holtan DW, Stabenfeldt GH, Hughes JP, Neely DP, Wahle WF. Source of Oestrogen in the Mare. Endocrinology 96, no. 2: 522–528.
    doi: 10.1210/endo-96-2-522google scholar: lookup
  19. Kalisch‐Smith JI, Simmons DG, Dickinson H, Moritz KM. Sexual Dimorphism in the Formation, Function and Adaptation of the Placenta. Placenta 54: 10–16.
    pubmed: 27979377
  20. Kibushi M, Kawate N, Kaminogo Y. Fetal Gender Prediction Based on Maternal Plasma Testosterone and Insulin‐Like Peptide 3 Concentrations at Midgestation and Late Gestation in Cattle. Theriogenology 86, no. 7: 1764–1773.
  21. Legacki EL, Ball BA, Corbin CJ. Equine Fetal Adrenal, Gonadal and Placental Steroidogenesis. Reproduction 154, no. 4: 445–454.
    doi: 10.1530/REP-17-0239pubmed: 28878092google scholar: lookup
  22. Livini M. Determination of Fetal Gender by Transrectal Ultrasound Examination: Field's Experience. AAEP Proceedings 56: 323–327.
  23. Mari G, Castagnetti C, Belluzzi S. Equine Fetal Sex Determination Using a Single Ultrasonic Examination Under Farm Conditions. Theriogenology 58, no. 6: 1237–1243.
    doi: 10.1016/S0093-691X(02)00943-3pubmed: 12240926google scholar: lookup
  24. Mcgladdery A. Equine Fetal Sex Determination. In Proceedings of the Annual Meeting of the Italian Association of Equine Veterinarians, 113–115.
  25. Merkt H, Moura JC, Jochle W. Gender Determination in Equine Fetuses Between Days 50 and 90 of Pregnancy. Journal of Equine Veterinary Science 19, no. 2: 90–94.
  26. Nakai R, Weng Q, Tanaka Y. Change in Circulating Follicle‐Stimulating Hormone, Luteinizing Hormone, Immunoreactive Inhibin, Progesterone, Testosterone and Estradiol‐17β in Fillies From Birth to 6 Months of Age. Journal of Equine Science 18, no. 3: 85–91.
    doi: 10.1294/jes.18.85pmc: PMC4013995pubmed: 24833982google scholar: lookup
  27. Oliveira RA, Yamim RS, Pivato I. Sexagem Fetal Em Equinos. Revista Brasileira De Reprodução Animal 38: 37–42.
  28. Ousey JC. Endocrinology of Pregnancy. In Equine Reproduction, edited by Mckinnon AO, Varner DD, Squires EL, and Vaala WE, 2nd ed., 2222–2233.
  29. Panarace M, Pellegrini RO, Basualdo MO. First Field Results on the Use of Stallion Sex‐Sorted Semen in a Large‐Scale Embryo Transfer Program. Theriogenology 81, no. 4: 520–525.
  30. Pashen RL, Lascombes FA, Darrow MD. The Application of Embryo Transfer to Polo Ponies in Argentina. Equine Veterinary Journal 15: 119–121.
  31. Renaudin CD, Gillis CL, Tarantal AF. Transabdominal Ultrasonographic Determination of Fetal Gender in the Horse During Mid‐Gestation. Equine Veterinary Journal 31, no. 6: 483–487.
  32. Rosenfeld CS. Sex‐Specific Placental Responses in Fetal Development. Endocrinology 156, no. 10: 3422–3434.
    pmc: PMC4588817pubmed: 26241064
  33. Salles MGF, Araújo AA. Corpo Lúteo Cíclico e Gestacional: Revisão. Revista Brasileira de Reprodução Animal 34, no. n.3: 185–194.
  34. Taveiros AW, Freitas Neto LM, Aguiar Filho CR. Utilização do Ultrassom Para Sexar Fetos Equinos da Raça Mangalarga Marchador Pela Visualização do Tubérculo Genital e da Genitália. Medicina Veterinária 2, no. 4: 35–40.

Citations

This article has been cited 0 times.