he Cut-Off Value for Classifying Active Italian Children Using the Corresponding National Version of the Physical Activity Questionnaire

The present study aimed to determine a cut-off value following the filling in of a questionnaire (PAQ-C-It) to identify active Italian children. One-hundred-twenty-nine primary school children (5 Piedmont schools; 47.3% female; mean age = 10 ± 1 years) wore an accelerometer (Actigraph wGT3X-BT) to objectively quantify individual moderate-to-vigorous physical activity during one week. Afterwards, the PAQ-C-It was filled in by participants. A ROC curve procedure was applied to obtain an active/non-active cut-off point. Spearman's correlation coefficient was also applied to establish the relationship between the two parameters. According to the ROC analysis, the PAQ-C-It cut-off point value is identifiable at >2.75 to indicate active children (area under the curve = 0.62; standard error = 0.05; p = 0.025; coefficient intervals = 0.518-0.716; sensitivity = 0.592, specificity = 0.382), determining that 65 participants (55%) were non-active (mean PAQ-C-It value = 2.3 ± 0.4; active mean PAQ-C-It value = 3.3 ± 0.4). Spearman's correlation coefficient results were significant but with a small effect size (rho = 0.214; p = 0.008). In conclusion, the present results suggest that the PAQ-C-It can be cautiously used as tool to practically classify active Italian children because of a non-solid relationship between respective accelerometer data and MVPA daily data

A RIKILT yeast estrogen bioassay (REA) for estrogen residue detection in urine of calves experimentally treated with 17ß-estradiol

17ß-Estradiol is one of the most powerful sex steroids illegally used in bovine production. The objective of this study was to evaluate the application and the specificity of the RIKILT yeast estrogen bioassay (REA) for the detection of molecules with estrogenic activities in the urine of calves experimentally treated with anabolics. Four groups of six calves each received an injection of 17ß-estradiol intramuscularly (group B), androsterone and gliburide (group A), and testosterone (group C) molecules at different dosage for 40 days. Group D was the control. The ability of the REA test to detect estrogenic activity in urine samples from all animals was assessed. All estrogen-treated animals (group B) showed as being positive up to 7 days after administration of the highest dosage of 17ß-estradiol, while the other three groups showed as being negative. The identity of estrogenic molecules in the urine of group B (17ß-estradiol, 17-estradiol) was confirmed by gas chromatography-mass spectrometry (GC/MS). This is the first time the REA test has been applied to detect 17ß-estradiol in the urine of calves treated with the hormone in vivo. The technique may offer an advantageous laboratory method for the veterinary surveillance of illegal steroid use

Application of absolute qPCR as a screening method to detect illicit 17β-oestradiol administration in male cattle

<div><p>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 <i>in vivo</i> 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 (<i>n</i> = 12), consisting of animals treated with four doses of 17β-oestradiol (5 mg week⁻¹ per animal); and group B (<i>n</i> = 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; <i>n</i> = 6) or remained untreated (group D; <i>n</i> = 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.</p> </div