…10.1016/j.meatsci.2012.05.009 [5] Doran E, Whittington F W, Wood J D and McGivan J D 2002 Cytochrome P450IIE1 (CYP2E1) is induced by skatole and this induction is…
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…10.1016/j.meatsci.2012.05.009 [5] Doran E, Whittington F W, Wood J D and McGivan J D 2002 Cytochrome P450IIE1 (CYP2E1) is induced by skatole and this induction is…
oler and centrifuged (2,000 × g for 15 min at 4°C) within eight hours of collection to separate serum. The serum was then aliquoted into 1.5ml Axygen® microcentrifuge tubes (Axygen Scientific, Corning, NY) at − 80°C. The assays for the biomarker analysis were completed six to nine months post-collection. PGE2 concentrations were measured using a commercial enzyme-linked immunosorbent assay (ELISA; catalog No. 514531; Cayman Chemical) following the method described by Giorgi et al., (2011). Briefly, serum samples were purified by adding ice-cold acetone (4x the serum volume), followed by incubation at − 20° C for 30 minutes and centrifugation (3,000 x g for 5 minutes). The supernatant was transferred to a 13 x 100-mm glass tube, evaporated using a CentriVap concentrator (Labconco), and reconstituted to the original serum volume with kit buffer. An aliquot of the reconstituted sample was derivatized with adjusted kit components and the manufacturer’s protocol were followed. Samples were analyzed in duplicate, and absorbance was measured at 405 nm following 60 minutes of development (SpectraMax i3; Molecular Devices). The mean PGE2 concentration of a reference sample used for repeatability assessment was 12.42 pg/ml (range: 9.90–15.60 pg/mL), with an inter-assay coefficient of variation of 8.61% (Merenda et al., 2022). Haptoglobin concentrations were determined using the Tridelta Phase® Haptoglobin Assay (catalog No. TP-801; Tridelta Development Ltd., Maynooth, Ireland). The assay utilized Haemoglobin (reagent 1), Chromogen (reagent 2), calibrator/sample diluent, and calibrator (microplate method). A standard curve was generated for each assay using haptoglobin standards at 2.5, 1.25, 0.625, 0.312, and 0 mg/mL. The Page 5/20 assay procedure involved
oler and centrifuged (2,000 × g for 15 min at 4°C) within eight hours of collection to separate serum. The serum was then aliquoted into 1.5ml Axygen® microcentrifuge tubes (Axygen Scientific, Corning, NY) at − 80°C. The assays for the biomarker analysis were completed six to nine months post-collection. PGE2 concentrations were measured using a commercial enzyme-linked immunosorbent assay (ELISA; catalog No. 514531; Cayman Chemical) following the method described by Giorgi et al., (2011). Briefly, serum samples were purified by adding ice-cold acetone (4x the serum volume), followed by incubation at − 20° C for 30 minutes and centrifugation (3,000 x g for 5 minutes). The supernatant was transferred to a 13 x 100-mm glass tube, evaporated using a CentriVap concentrator (Labconco), and reconstituted to the original serum volume with kit buffer. An aliquot of the reconstituted sample was derivatized with adjusted kit components and the manufacturer’s protocol were followed. Samples were analyzed in duplicate, and absorbance was measured at 405 nm following 60 minutes of development (SpectraMax i3; Molecular Devices). The mean PGE2 concentration of a reference sample used for repeatability assessment was 12.42 pg/ml (range: 9.90–15.60 pg/mL), with an inter-assay coefficient of variation of 8.61% (Merenda et al., 2022). Haptoglobin concentrations were determined using the Tridelta Phase® Haptoglobin Assay (catalog No. TP-801; Tridelta Development Ltd., Maynooth, Ireland). The assay utilized Haemoglobin (reagent 1), Chromogen (reagent 2), calibrator/sample diluent, and calibrator (microplate method). A standard curve was generated for each assay using haptoglobin standards at 2.5, 1.25, 0.625, 0.312, and 0 mg/mL. The Page 5/20 assay procedure involved
remaining samples, its level was undetectable. The highest average androsterone content was determined in the backfat of boars from group E1, with an undetectable content in one sample. The lowest average was found in group E2, with three undetectable samples. Based on the results obtained using the Heracles II electronic nose, various volatile compounds was identified in both meat and backfat, with differences observed between the experimental groups (Tables 8 and 9). Table 8. Volatile compound profile in meat. Possibly Matching Chemical Groups of Sensory Groups Compounds IR KMXT-5 * Compounds Descriptors C E1 E2 methyl formate 372 ester agreeable X X X ethanol 450 alcohol alcoholic X X X propanal 491 aldehyde earthy X X X 1-propanol 521 alcohol alcoholic X X X 2-mercaptoethanol 558 thiol sulphurous X X 2-methylfuran 584 furan burnt X but-(2)-enal 635 aldehyde green X X X 3-methylbutanal 652 aldehyde fatty X X methyl isobutyrate 665 ester fruity X 2-ethyl furan 704 furan acidic X X X methyl butanorate 734 ester ester X X propylenglycol 754 alcohol alcoholic X 2-methylpentanal 762 aldehyde earthy X X X 2,3-butanediol 790 diol bitter X octane 825 alkane alkane X e-2-hexen-1-ol 848 alcohol green X 1-hexanol 876 alcohol fatty X X pentanoic acid 905 acid rancid X X X 5-methylfurfural 956 furan acidic X 2-(2- 1001 ether mild X X ethoxyethoxy)ethanol pinene 1003 terpen hay X heptyl mercaptan 1026 thiol sulphurous X benzeneacetaldehyde 1028 aldehyde grassy X X 1,8-cineole 1046 terpene herbaceous X 2-propionylpyrrole 1049 pyrrole roast X undecane 1076 alkane alkane X gamma-terpinene 1078 terpen etheral X X p-menthatriene 1131 terpene woody X 2,6-dimethoxy-phenol 1203 phenol phenolic X (e, e)-2,4-nonadienal 1208 aldehyde cereal X decanal 1229 aldehyde fatty X X indole 1295
lable, pre-treatment data were included as a co-variate. Baseline data were defined as the latest measurement of a given variable before treatment was administered. The statistical model included treatment, sex (except for parameters only measured in one sex), and treatment-by-sex interactions as fixed effects, while the repeated measures ANOVA model included treatment, sex and time (and their 2-way and 3-way interac- tions) as fixed effects. Data were compared either within or across sex depending on observed interactions. Models were selected using backwards elimination where 3-way and 2-way interaction terms with the highest p-Value were sequentially dropped from the model. The process was performed hierarchically starting with the 3-way interaction terms using a threshold p-Value of 0.1, as required by regulatory guidelines. Model selection was stopped when any of the following occurred: (i) all interaction terms at the highest level had a p-Value < 0.1; (ii) no interaction terms remained in the model. The statistical model was therefore: Parameter = Baseline_variables + Treatment + Sex + Time + Treatment:Sex + Treat- ment:Time + Sex:Time + Treatment:Sex:Time + Residual_Error Least squares means were compared at a significance level of p < 0.1 in the first instance apart from treatment by sex by time point, which was compared at p < 0.05 in the first instance. For the repeated measures models, most residual error patterns were selected based on a comparison of Akaike Information Criteria (AIC) output for models containing an unstructured (UN), autoregressive (AR; 1), variance components (VC) and independent residual error pattern (with the lowest AIC preferred among the possible covariance matrices). The denominator degrees of freedom in the covariance pattern
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eting interests The authors declare no competing interests. Additional information Supplementary Information The online version contains supplementary material available at https://doi .org/1 0.10 38/s41598- 025-91551-6. Correspondence and requests for materials should be addressed to P.H.E.T. Reprints and permissions information is available at www.nature.com/reprints. Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http:/ /creativec ommo ns.org /licenses/
eting interests The authors declare no competing interests. Additional information Supplementary Information The online version contains supplementary material available at https://doi .org/1 0.10 38/s41598- 025-91551-6. Correspondence and requests for materials should be addressed to P.H.E.T. Reprints and permissions information is available at www.nature.com/reprints. Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http:/ /creativec ommo ns.org /licenses/