Integrated analytical approach in veal calves administered the anabolic androgenic steroids boldenone and boldione: urine and plasma kinetic profile and changes in plasma protein expression

Abstract

Surveillance of illegal use of steroids hormones in cattle breeding is a key issue to preserve human health. To this purpose, an integrated approach has been developed for the analysis of plasma and urine from calves treated orally with a single dose of a combination of the androgenic steroids boldenone and boldione. A quantitative estimation of steroid hormones was obtained by LC-APCI-Q-MS/MS analysis of plasma and urine samples obtained at various times up to 36 and 24 h after treatment, respectively. These experiments demonstrated that boldione was never found, while boldenone alpha- and beta-epimers were detected in plasma and urine only within 2 and 24 h after drug administration, respectively. Parallel proteomic analysis of plasma samples was obtained by combined 2-DE, MALDI-TOF-MS and muLC-ESI-IT-MS/MS procedures. A specific protein, poorly represented in normal plasma samples collected before treatment, was found upregulated even 36 h after hormone treatment. Extensive mass mapping experiments proved this component as an N-terminal truncated form of apolipoprotein A1 (ApoA1), a protein involved in cholesterol transport. The expression profile of ApoA1 analysed by Western blot analysis confirmed a significant and time dependent increase of this ApoA1 fragment. Then, provided that further experiments performed with a growth-promoting schedule will confirm these preliminary findings, truncated ApoA1 may be proposed as a candidate biomarker for steroid boldenone and possibly other anabolic androgens misuse in cattle veal calves, when no traces of hormones are detectable in plasma or urine.

Feeding milk replacer instead of whole milk affects blood plasma proteome and lipid profile in preruminant calves

“…The presence of these proteins in the blood plasma of cattle was previously shown in our study as well as in the works of other authors (Wait et al 2002, D'Ambrosio et al 2005, Draisci et al 2007, Yang et al 2012.…”

“…There have been numerous studies investigating the effects of different factors on plasma proteome changes in cattle; for example, Yang et al (2012) found significant changes in plasma acute phase proteins (APPs) expression, using two-dimensional electrophoresis (2-DE), in cows which had developed post-partum subclinical mastitis. An attempt has also been made to evaluate the functional molecular plasma biomarkers for the detection of growth-promoting substances which are illegally used in veal calves (Draisci et al 2007). In our previous study, the dynamic changes in plasma proteome expression pattern from birth to the seventh day of the life of calves which were fed dams colostrum and milk were observed (Herosimczyk et al 2013).…”

Blood plasma protein and lipid profile changes in calves during the first week of life

“…Proteomic tools have also been employed to reveal molecular serum/plasma biomarkers for the detection of growth promoting substances that are illegally used in the veal calves. Recently, two-dimensional electrophoresis (2-DE), matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF-MS) and liquid chromatography-hybrid ion trap time of flight mass spectrometry (μLC-IT-TOF/MS) have been successfully applied to the global analysis of plasma from calves treated orally with the androgenic steroids such as boldenone and boldione (Draisci et al 2007). Draisci et al (2007) showed that single dose of the boldenone resulted in the overexpression of the N-terminal truncated form of apolipoprotein A-I and thus may be considered as a candidate biomarker for the anabolic androgens treatment.…”

“…Recently, two-dimensional electrophoresis (2-DE), matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF-MS) and liquid chromatography-hybrid ion trap time of flight mass spectrometry (μLC-IT-TOF/MS) have been successfully applied to the global analysis of plasma from calves treated orally with the androgenic steroids such as boldenone and boldione (Draisci et al 2007). Draisci et al (2007) showed that single dose of the boldenone resulted in the overexpression of the N-terminal truncated form of apolipoprotein A-I and thus may be considered as a candidate biomarker for the anabolic androgens treatment. Nevertheless, in the available literature there is a lack of information concerning other age-dependent and diet-dependent plasma proteome changes that occur in response to dynamic adaptation processes, during the early postnatal period.…”

Fragmentation of Human Erythrocyte Actin following Exposure to Hypoxia

“…Isoelectrofocusing was conducted using a Multiphor system (GE Healthcare) for the first-dimension analysis, and an Ettan Dalt Six apparatus for SDS-PAGE (GE Healthcare) was used according to the manufacturer’s instructions. Focused strips were equilibrated according to a standard procedure and then applied to 12.5% SDS polyacrylamide gels [18,19]. After staining with colloidal Coomassie blue, the gels were scanned with a 2D image scanner (GE Healthcare).…”