Early Diagnostics of Freemartinism in Polish Holstein-Friesian Female Calves

Abstract

Simple SummaryFreemartinism is the most common type of gender developmental disorder, resulting in infertility of heifers from multiple-sex twin pregnancies. The frequency of this syndrome is related to the frequency of multiple pregnancies, the number of which has significantly increased in dairy cattle populations (HF). Therefore, rapid diagnostics is necessary to enable early elimination of heifers with freemartinism from breeding. The aim of the study was to compare and identify the best method for early identification of freemartinism. The use of cytogenetic and molecular methods (PCR, short tandem repeats (STRs), real-time PCR) allowed us to conclude that molecular methods are more effective and guarantee fast and precise diagnosis. An additional advantage of molecular methods is the easy way to collect test material, which can be frozen, unlike blood samples for cytogenetic analysis, which must be fresh and delivered within 24 h to the laboratory, which generates further costs.AbstractFreemartinism in females born from heterosexual multiple pregnancies is characterized by the presence of XX/XY cell lines due to the formation of a shared blood system by anastomoses between fetal membranes of co–twins and leads to disturbed development of the reproductive system, including infertility. The aim of this study was to estimate the most precise and effective diagnostic method, especially useful for early identification of freemartinism in young female calves. The cytomolecular evaluation results of 24 Holstein-Friesian heifers from heterosexual twins was verified by molecular techniques: PCR, short tandem repeats (STRs), and relative quantitative PCR. The molecular analyses have been found to be a more efficient testing strategy, with a higher diagnostic success rate than karyotype evaluation. In 21 heifers, leucocyte chimerism determined by the 60, XX/60, XY karyotype was revealed—the proportion of the 60, XY male cell line in individual animals was in the range of 4–66%. In three cases, a normal karyotype 60, XX was identified, which indicates that anastomoses did not occur in 12.5% of studied twins and suggests that these potentially fertile heifers can be qualified for further breeding. The precise and early identification of freemartinism can be the basis for guidelines and selection recommendations concerning the reproductive performance of heifers born from heterosexual multiple pregnancies.

Disorders of sexual development in small ruminants

“…10 In cattle, freemartinism affects > 90% of heifers born from a heterosexual twin pregnancy 3 whereas in sheep, the incidence is as low as 1-5% in male-bearing multiple pregnancies. 6,10 The more common development of vascular placental anastomosis between fetuses is responsible for the high incidence of freemartin in cattle 11 but this phenomenon is less frequent in small ruminants. 2,3 It has been suggested that in highly prolific sheep breeds, male-bearing pregnancies with 4 or 5 lambs might present an increased risk for freemartinism, potentially due to increased placental contact between fetuses.…”

Assessment of the genomic stability of calves obtained from artificial insemination and OPU/IVP in vitro fertilization

“…Some researchers report that the in vitro method of acquiring calves entails complications, including heavier embryos resulting in more difficult deliveries (Behboodi et al, 1995;Breukelman et al, 2005;Kruip & den Haas, 1997;Numabe et al, 2000), slower development (Max, 2016), higher oxygen consumption, lipid accumulation, DNA methylation and mixoploidy (Hansen, 2020a(Hansen, , 2020bViuff et al, 1999), as well as chromosomal anomalies (Block et al, 2003;Bonilla et al, 2013;Iannuzzi et al, 2021;Kozubska-Sobocińska et al, 2019;van Wagtendonk-de Leeuw et al, 1998). Viuff et al (1999), Dieleman et al (2002), Tšuiko et al (2017), Catteeuw (2018) and Hansen (2020aHansen ( , 2020b found that chromosomal anomalies were more common in in vitro embryos than in those obtained in vivo.…”

A Disorder of Sex Development in a Holstein–Friesian Heifer with a Rare Mosaicism (60,XX/90,XXY): A Genetic, Anatomical, and Histological Study

“…The affected females are sterile due to their underdeveloped reproductive systems. In dairy cattle breeds, an increased trend of twin pregnancies is observed, and this type of DSD is thus considered a serious health and breeding problem [ 4 , 5 ].…”