title: Application of absolute qPCR as a screening method to detect illicit 17β-oestradiol administration in male cattle.
abstract: It has been previously demonstrated that the progesterone receptor gene is up-regulated in the sex accessory glands of pre-pubertal and adult male bovines after 17β-oestradiol treatment. In the present study, a qualitative screening method was optimised to detect 17β-oestradiol treatment using absolute quantification by qPCR of the progesterone receptor gene to determine the amount of gene expression in bulbo-urethral glands. An external standard curve was generated and developed with TaqMan® technology. Based on two in vivo experiments, the decision limit CCα, sensitivity and specificity of this screening method were established. Trial 1 consisted of 32 Friesian veal calves divided into two groups: group A (n = 12), consisting of animals treated with four doses of 17β-oestradiol (5 mg week(-1) per animal); and group B (n = 20), consisting of control animals. Trial 2 was performed on 26 Charolaise beef cattle that either received five doses of 17β-oestradiol (group C; 20 mg week(-1) per animal; n = 6) or remained untreated (group D; n = 20). Further, progesterone receptor gene expression was evaluated in beef and veal calves for human consumption. A specific CCα on 20 Piedmontese control beef cattle was calculated to include these animals in a field investigation. Five out of 190 beef cattle and 26 out of 177 calves tested expressed the progesterone receptor gene above their respective CCα and they were classified as being suspected of 17β-oestradiol treatment. Additionally, 58% of veal calves that tested suspect via qPCR exhibited histological lesions of the bulbo-urethral gland tissue, which are typical of oestrogen administration and are consistent with hyperplasia and metaplasia of the glandular epithelium.
authors.affiliation: Dipartimento di Patologia Animale, Facoltà di Medicina Veterinaria, Università degli Studi di Torino, Turin, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Uslenghi
authors.forename: S
authors.initials: S
authors.lastname: Divari
authors.forename: F T
authors.initials: FT
authors.lastname: Cannizzo
authors.forename: R
authors.initials: R
authors.lastname: De Maria
authors.forename: F
authors.initials: F
authors.lastname: Spada
authors.forename: C
authors.initials: C
authors.lastname: Mulasso
authors.forename: M
authors.initials: M
authors.lastname: Pezzolato
authors.forename: E
authors.initials: E
authors.lastname: Bozzetta
authors.forename: A
authors.initials: A
authors.lastname: Attucci
authors.forename: P
authors.initials: P
authors.lastname: Giorgi
authors.forename: B
authors.initials: B
authors.lastname: Biolatti
mesh_terms: D000818:Animals; D002030:Bulbourethral Glands / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002417:Cattle; D004350:Drug Residues / Q000032:analysis; D004958:Estradiol / Q000008:administration & dosage*; D005506:Food Contamination / Q000032:analysis*; D006801:Humans; D008297:Male; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011980:Receptors, Progesterone / Q000235:genetics*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D011980:Receptors, Progesterone; D004958:Estradiol