Regucalcin expression profiles in veal calf testis: validation of histological and molecular tests to detect sex steroids illicit administration.
Immunohistochemistry
FFPE tissues
Growth promoters
Real Time PCR
Regucalcin
Journal
PeerJ
ISSN: 2167-8359
Titre abrégé: PeerJ
Pays: United States
ID NLM: 101603425
Informations de publication
Date de publication:
2021
2021
Historique:
received:
27
05
2020
accepted:
13
01
2021
entrez:
1
3
2021
pubmed:
2
3
2021
medline:
2
3
2021
Statut:
epublish
Résumé
Sex steroids administration in meat producing animals is forbidden within the EU to preserve consumers' safety, but continuous monitoring to identify resurgence of their misuse is needed. Among biomarkers related to sex steroids abuse in veal calves the regucalcin (RGN) mRNA perturbations in testis have been described in RNA Formalin fixed paraffin embedded (FFPE) testis samples, chosen being the most effective preservation strategy adopted by histological NRCPs and allowing easier retrospective analysis if required by legal disputes, were analyzed from veal calves treated with nandrolone, 17β-estradiol and a cocktail of the two hormones. RGN levels were determined by quantitative Real Time PCR and Immunohistochemistry assays. Test performances were assessed and compared by multiple ROC curves. Both tests resulted sensitive and specific, allowing to enrich, in future field investigation, novel integrated diagnostic protocols needed to unveil sex steroid abuse. Developed RT-qPCR and IHC methods confirmed RGN as a useful and robust biomarker to detect illegal administration of sex steroid hormones in veal calves. The developed methods, successfully applied to ten years old FFPE blocks, could allow both retrospective analysis, when supplementary investigations are requested by authorities, and future implementation of current NRCPs.
Sections du résumé
BACKGROUND
BACKGROUND
Sex steroids administration in meat producing animals is forbidden within the EU to preserve consumers' safety, but continuous monitoring to identify resurgence of their misuse is needed. Among biomarkers related to sex steroids abuse in veal calves the regucalcin (RGN) mRNA perturbations in testis have been described in RNA
METHODS
METHODS
Formalin fixed paraffin embedded (FFPE) testis samples, chosen being the most effective preservation strategy adopted by histological NRCPs and allowing easier retrospective analysis if required by legal disputes, were analyzed from veal calves treated with nandrolone, 17β-estradiol and a cocktail of the two hormones. RGN levels were determined by quantitative Real Time PCR and Immunohistochemistry assays. Test performances were assessed and compared by multiple ROC curves.
RESULTS
RESULTS
Both tests resulted sensitive and specific, allowing to enrich, in future field investigation, novel integrated diagnostic protocols needed to unveil sex steroid abuse.
DISCUSSION
CONCLUSIONS
Developed RT-qPCR and IHC methods confirmed RGN as a useful and robust biomarker to detect illegal administration of sex steroid hormones in veal calves. The developed methods, successfully applied to ten years old FFPE blocks, could allow both retrospective analysis, when supplementary investigations are requested by authorities, and future implementation of current NRCPs.
Identifiants
pubmed: 33643712
doi: 10.7717/peerj.10894
pii: 10894
pmc: PMC7899017
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e10894Informations de copyright
©2021 Benedetto et al.
Déclaration de conflit d'intérêts
The authors declare there are no competing interests.
Références
BMC Bioinformatics. 2018 Oct 15;19(Suppl 10):357
pubmed: 30367588
Chem Pharm Bull (Tokyo). 1978 Jun;26(6):1915-8
pubmed: 699201
Inhal Toxicol. 2014 Nov;26(13):811-28
pubmed: 25264934
Nucleic Acids Res. 2012 Apr;40(7):e51
pubmed: 22228834
BMC Mol Biol. 2007 Dec 21;8:118
pubmed: 18154675
Eur J Intern Med. 2011 Jun;22(3):230-4
pubmed: 21570638
Food Chem. 2020 May 15;312:126061
pubmed: 31891886
Biomol Detect Quantif. 2015 Sep 26;6:4-12
pubmed: 27077042
Mol Hum Reprod. 2011 Nov;17(11):669-78
pubmed: 21593214
Sci Rep. 2018 Apr 20;8(1):6351
pubmed: 29679021
EFSA J. 2013 Jun 27;11(6):3265
pubmed: 32313569
Sci Rep. 2016 Feb 22;6:21418
pubmed: 26898768
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2016;33(1):60-5
pubmed: 26482144
Biopreserv Biobank. 2015 Jun;13(3):200-6
pubmed: 26035010
Nucleic Acids Res. 2003 Jul 1;31(13):3406-15
pubmed: 12824337
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2014;31(5):833-8
pubmed: 24588558
Stat Med. 2010 Dec 10;29(28):2905-19
pubmed: 20862668
PLoS One. 2014 Nov 21;9(11):e113950
pubmed: 25415588
Mol Hum Reprod. 2012 Apr;18(4):161-70
pubmed: 22121208
J Agric Food Chem. 2017 Jun 14;65(23):4866-4874
pubmed: 28525721