Equine fetal genotyping via aspiration of yolk-sac fluid at 22–28 days of gestation

A. M. Ripley, M. C.T. Penedo, R. A. Grahn, E. V. Martinez de Andino, S. R. Walbornn, R. Serafini, C. C. Love, K. Hinrichs

Research output: Contribution to journalArticle

Abstract

Fetal genotyping has important applications in the horse, but currently necessitates embryo recovery and biopsy. We investigated whether fetal genotyping could be performed on yolk-sac fluid recovered from pregnant mares via transvaginal aspiration. Fluid was collected before Day 30 to provide results before establishment of the endometrial cups (Day 37). Genotyping and assessment of maternal DNA contamination was performed by analyzing histograms of PCR results for 19 loci. In Exp. 1, mares underwent yolk-sac aspiration on Days 22–28 of gestation. Fluid (0.56–1.02 mL) was recovered from five of seven mares. Four of the five mares maintained pregnancy. One pregnancy was electively terminated at Day 75; the other three mares delivered healthy foals. Extraction of DNA from the fluid sample followed by direct PCR allowed the highest rate of determination of fetal alleles. Fetal genotype was correctly determined in three samples, and for 14/19 alleles in one sample. In Exp. 2, we evaluated whether recovery of more fluid (up to 1.6 mL), and fractionation of the sample, would minimize maternal DNA contamination. One of four mares maintained pregnancy. Evaluation at informative loci showed no difference in maternal contamination among fractions. We determined that mares can maintain pregnancy after aspiration of yolk-sac fluid, and that fetal genotype can be accurately determined from the sample obtained. Further work is needed on factors affecting maintenance of pregnancy after the procedure. The ability to access the yolk sac in early pregnancy opens the door to novel potential clinical and research applications.

Original languageEnglish (US)
Pages (from-to)34-40
Number of pages7
JournalTheriogenology
Volume142
DOIs
StatePublished - Jan 15 2020

Fingerprint

Yolk Sac
yolk sac
genotyping
mares
Horses
pregnancy
horses
Pregnancy
DNA Contamination
Mothers
Alleles
Genotype
Pregnancy Maintenance
DNA
sampling
Polymerase Chain Reaction
alleles
loci
genotype
fluids

Keywords

  • Allantocentesis
  • Amniocentesis
  • Equine
  • Genetic diagnosis

ASJC Scopus subject areas

  • Small Animals
  • Food Animals
  • Animal Science and Zoology
  • Equine

Cite this

Equine fetal genotyping via aspiration of yolk-sac fluid at 22–28 days of gestation. / Ripley, A. M.; Penedo, M. C.T.; Grahn, R. A.; Martinez de Andino, E. V.; Walbornn, S. R.; Serafini, R.; Love, C. C.; Hinrichs, K.

In: Theriogenology, Vol. 142, 15.01.2020, p. 34-40.

Research output: Contribution to journalArticle

Ripley, AM, Penedo, MCT, Grahn, RA, Martinez de Andino, EV, Walbornn, SR, Serafini, R, Love, CC & Hinrichs, K 2020, 'Equine fetal genotyping via aspiration of yolk-sac fluid at 22–28 days of gestation', Theriogenology, vol. 142, pp. 34-40. https://doi.org/10.1016/j.theriogenology.2019.09.012
Ripley, A. M. ; Penedo, M. C.T. ; Grahn, R. A. ; Martinez de Andino, E. V. ; Walbornn, S. R. ; Serafini, R. ; Love, C. C. ; Hinrichs, K. / Equine fetal genotyping via aspiration of yolk-sac fluid at 22–28 days of gestation. In: Theriogenology. 2020 ; Vol. 142. pp. 34-40.
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AB - Fetal genotyping has important applications in the horse, but currently necessitates embryo recovery and biopsy. We investigated whether fetal genotyping could be performed on yolk-sac fluid recovered from pregnant mares via transvaginal aspiration. Fluid was collected before Day 30 to provide results before establishment of the endometrial cups (Day 37). Genotyping and assessment of maternal DNA contamination was performed by analyzing histograms of PCR results for 19 loci. In Exp. 1, mares underwent yolk-sac aspiration on Days 22–28 of gestation. Fluid (0.56–1.02 mL) was recovered from five of seven mares. Four of the five mares maintained pregnancy. One pregnancy was electively terminated at Day 75; the other three mares delivered healthy foals. Extraction of DNA from the fluid sample followed by direct PCR allowed the highest rate of determination of fetal alleles. Fetal genotype was correctly determined in three samples, and for 14/19 alleles in one sample. In Exp. 2, we evaluated whether recovery of more fluid (up to 1.6 mL), and fractionation of the sample, would minimize maternal DNA contamination. One of four mares maintained pregnancy. Evaluation at informative loci showed no difference in maternal contamination among fractions. We determined that mares can maintain pregnancy after aspiration of yolk-sac fluid, and that fetal genotype can be accurately determined from the sample obtained. Further work is needed on factors affecting maintenance of pregnancy after the procedure. The ability to access the yolk sac in early pregnancy opens the door to novel potential clinical and research applications.

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