Search Results (40)

The nuclear receptors pregnane X receptor (PXR), constitutive androstane receptor (CAR), and farnesoid X receptor (FXR) regulate the hepatic metabolism of androstenone, a testicular steroid that accumulates in the fat of intact male pigs and causes boar taint. This study evaluated natural product-derived compounds and conventional agonists targeting these nuclear receptors for their effects on androstenone metabolism in primary hepatocytes from slaughter-weight boars, to assess their potential as treatments for boar taint. Cells were incubated with natural products, conventional agonists, or dimethyl sulfoxide (DMSO; control), then being treated with androstenone. Culture media and cells were analyzed to assess changes in androstenone metabolism and gene expression. UGT1A6 was upregulated by treatments targeting both PXR and CAR and downregulated by FXR agonists. Additionally, PGC1α and NR2F1 were downregulated by compounds targeting PXR/CAR, while FXR and NR0B2 were upregulated and HNF4α downregulated by treatments acting on FXR. The natural products diallyl sulfide (DAS) and (Z)-guggulsterone (GUG) increased overall androstenone metabolism (DAS, GUG) and the production of Phase I androstenol metabolites (DAS), but only in hepatocyte culture replicates that responded positively to these treatments. Although gene expression was similar between positive-response and negative/non-responsive replicates following treatments, negative/non-responsive replicates for several treatments had higher basal expression of UGT2B31, UGT2A1, and SIRT1 and lower basal expression of FXR, PXR, and NR0B1 compared to positive-response replicates. These findings suggest that DAS and GUG may be promising treatments for boar taint, specifically in animals with lower basal rates of androstenone metabolism and higher expression of key nuclear receptors. Full article

The utility of four mineral adsorbents as potential feed additives to bind the boar taint compounds, androstenone and skatole, was assessed with an in vitro system. The adsorbents were bentonite (BNT), diatomaceous earth (DE), spent filter aid (SFA) and hydrated sodium–calcium aluminosilicate (HSCAS), with activated charcoal (AC) as a positive control. The binding capacity (B_max) and binding affinity (K) of androstenone (AND), estrone (E1), estrone sulfate (E1S), and skatole were estimated using the modified Michaelis–Menten kinetics. The Langmuir and Freundlich isotherm models were also used to assess the adsorption behaviour. The B_max values with AND were 77.7 ± 1.12%, 71.9 ± 1.93%, 55.0 ± 7.85%, and 69.5 ± 1.39% for BNT, DE, SFA, and HSCAS, respectively, with no differences in the binding affinity K (p > 0.05). All the mineral adsorbents had very low binding with E1S. SFA bound skatole with a B_max of 89.9 ± 1.09%, while the B_max values for skatole binding by BNT, DE and HCAS were approximately 15%. Most adsorbent–adsorbate complexes fit best with the Freundlich isotherm model. We conclude that all four mineral adsorbents bound androstenone, but not E1S, and only SFA effectively bound skatole. This suggests that SFA may act as a selective dietary binding agent to control boar taint, but further research using animal models is needed to explore the utility and selectivity of these adsorbents as feed additives to control boar taint. Full article

Olfactory receptors (ORs) are members of the transmembrane G protein-coupled receptor superfamily, playing a crucial role in odor recognition, which further mediates crucial biological processes in mammals. In sows, androstenone can trigger sexual behaviors through olfaction, but the underlying mechanism remains to be explored. To efficiently and accurately screen pig olfactory receptors responding to androstenone and the key structure determinant, we adapted the high-throughput RNA-seq strategy to screen the altered genes upon androstenone treatment in the olfactory epithelium of pigs, yielding 1397 downregulated genes. Of which, 15 OR genes and 49 OR-like genes were candidate androstenone-responsive genes, and 5 ORs (OR2D2, OR8D1, OR8D2, OR10Z1 and OR7D4) were proven as responsible for androstenone-mediated olfaction in vitro. Among the five ORs, pig OR7D4 has the highest level of androstenone response. To further find the structural determinant, we performed ligand-binding cavity analysis on pig OR7D4 with androstenone, predicted seven potential structural sites and further confirmed that F178 and T203 are the key sites for recognizing androstenone. Nevertheless, the natural non-synonymous mutation M133V (rs696400829) of pig OR7D4 was proven to significantly impair the respondence to androstenone. This is the first time the ORs responding to androstenone in pigs and the key structural determinant of pig OR7D4 were identified, which highlights the significance of investigating the role of OR7D4 in pig reproduction performance in the future. Full article

Testicular steroids can alter the activity and expression of enzymes within the liver and may influence the metabolism of skatole and androstenone, which are responsible for boar taint. Plasma levels of estrone sulfate (E₁S) are indicative of the steroidogenic capacity of the boar and are variable between animals of similar live weights at slaughter. This study aimed to characterize the relationship between steroidogenic capacity and the metabolism of boar taint compounds by relating plasma E₁S levels at slaughter weight to the expression levels of genes regulating the metabolism of androstenone and skatole, along with their respective metabolite profiles. RT-qPCR was used to evaluate gene expression in the liver. Hepatocytes were also isolated and treated with androstenone or skatole, with metabolite levels in the incubation media quantified by high-performance liquid chromatography. Plasma E₁S levels ranged from 2.2–108.5 ng/mL and were positively correlated with overall skatole metabolism (p = 0.038), the production of metabolites 3-methyloxindole (p = 0.026) and 3-hydroxy-3-methyloxindole (p = 0.036), and expression levels of key genes involved in skatole metabolism, specifically CYP2C33 (p = 0.0042), CYP2C49 (p = 0.022), and CYB5R1 (p = 0.017). There was no association between androstenone metabolism and plasma E₁S concentrations; however, there was evidence of possible co-regulation amongst genes involved in the metabolism of androstenone, skatole, and estrogens. These findings indicate that steroidogenic capacity is related to the rate of skatole, but not androstenone metabolism, in slaughter-weight boars. Full article

Boar taint, an unfavorable odor in the meat of intact male pigs, is caused primarily by the accumulation of two compounds: androstenone and skatole. This multifactorial trait is regulated by numerous dietary, management and genetic factors. At the mechanistic level, there are many genes known to be involved in boar taint metabolism. Cytochrome P450 2E1 (CYP2E1) impacts boar taint through the phase I metabolism of skatole. The aim of this study was to identify single-nucleotide polymorphisms (SNPs) within the CYP2E1 gene promoter and explore their relationship with the expression of CYP2E1 mRNA and protein. Sequencing of the promoter region using pools of genomic DNA identified seven promoter region SNPs at −159, −586, −1693, −1806, −2322, −2369 and −2514 bp upstream of the ATG start site. Genomic DNA was obtained from 65 boars from the three major swine breeds: Duroc, Landrace and Yorkshire, and individual animals were genotyped for the identified SNPs. RNA was isolated from liver tissue and quantitative PCR was performed to measure CYP2E1 gene expression, while levels of CYP2E1 protein in liver were measured by Western blotting. Significant within-breed variation in CYP2E1 protein and mRNA expression was observed, indicating significant differences in gene expression among individuals. However, levels of CYP2E1 mRNA and protein were not significantly correlated. Two SNPs within the promoter were significantly associated with CYP2E1 mRNA expression, but not with protein expression. This study provides evidence of additional mutations affecting the gene expression of CYP2E1 and suggests that factors that affect the differences in translation of CYP2E1 mRNA may also be important in affecting skatole metabolism. Full article

Surgical castration can effectively avoid boar taint and improve pork quality by removing the synthesis of androstenone in the testis, thereby reducing its deposition in adipose tissue. The expression of genes involved in testis-derived hormone metabolism was altered following surgical castration, but the upstream regulatory factors and underlying mechanism remain unclear. In this study, we systematically profiled chromatin accessibility and transcriptional dynamics in liver tissue of castrated and intact full-sibling Yorkshire pigs. First, we identified 897 differentially expressed genes and 6864 differential accessible regions (DARs) using RNA- and ATAC-seq. By integrating the RNA- and ATAC-seq results, 227 genes were identified, and a significant positive correlation was revealed between differential gene expression and the ATAC-seq signal. We constructed a transcription factor regulatory network after motif analysis of DARs and identified a candidate transcription factor (TF) SP1 that targeted the HSD3B1 gene, which was responsible for the metabolism of androstenone. Subsequently, we annotated DARs by incorporating H3K27ac ChIP-seq data, marking 2234 typical enhancers and 245 super enhancers involved in the regulation of all testis-derived hormones. Among these, four typical enhancers associated with HSD3B1 were identified. Furthermore, an in-depth investigation was conducted on the androstenone-related enhancers, and an androstenone-related mutation was identified in a newfound candidatetypical enhancer (andEN) with dual-luciferase assays. These findings provide further insights into how enhancers function as links between phenotypic and non-coding area variations. The discovery of upstream TF and enhancers of HSD3B1 contributes to understanding the regulatory networks of androstenone metabolism and provides an important foundation for improving pork quality. Full article

The extraction of boar taint compounds from pork fat samples was performed under various temperature (150, 300 and 450 °C) and atmosphere (air, nitrogen and reduced pressure) conditions. This aimed at understanding which conditions allow the greatest extractions of indole, skatole and androstenone (present in backfat in low concentrations) while limiting the presence of other VOCs in the headspace of heated fat (interfering with correct VOC-based detection of boar taint compounds). Indole and skatole were extracted in the greatest concentrations when heating backfat at 450 °C under reduced pressure, while androstenone was highest when heating at 300 °C under reduced pressure. Oxidation products were most abundant under air conditions, nitrogenated products appeared in the presence of a nitrogen-enriched atmosphere, and lastly, molecules intrinsic to boar fat saw their headspace concentration increase with reduced pressure. The combination of 450 °C and reduced pressure atmosphere was suggested for the heating of backfat prior to detection with analytical methods and to complement the current sensory analysis. Full article

With a perspective future ban on surgical castration in Europe, selecting pigs with reduced ability to accumulate boar taint (BT) compounds (androstenone, indole, skatole) in their tissues seems a promising strategy. BT compound concentrations were quantified in the adipose tissue of 1075 boars genotyped at 29,844 SNPs. Traditional and SNP-based breeding values were estimated using pedigree-based BLUP (PBLUP) and genomic BLUP (GBLUP), respectively. Heritabilities for BT compounds were moderate (0.30–0.52). The accuracies of GBLUP and PBLUP were significantly different for androstenone (0.58 and 0.36, respectively), but comparable for indole and skatole (~0.43 and ~0.47, respectively). Several SNP windows, each explaining a small percentage of the variance of BT compound concentrations, were identified in a genome-wide association study (GWAS). A total of 18 candidate genes previously associated with BT (MX1), reproduction traits (TCF21, NME5, PTGFR, KCNQ1, UMODL1), and fat metabolism (CTSD, SYT8, TNNI2, CD81, EGR1, GIPC2, MIGA1, NEGR1, CCSER1, MTMR2, LPL, ERFE) were identified in the post-GWAS analysis. The large number of genes related to fat metabolism might be explained by the relationship between sexual steroid levels and fat deposition and be partially ascribed to the pig line investigated, which is selected for ham quality and not for lean growth. Full article

Selecting pigs with reduced ability to accumulate boar taint (BT) compounds in their tissues is an alternative to male surgical castration. As the majority of slaughter pigs are crossbred, before selecting against BT in purebreds, it is essential to consider possible impacts on commercial traits in crossbreds. This study estimated the genetic correlations between BT compound levels measured in 1115 purebred pigs and carcass and ham quality traits collected in 26,577 crossbred Italian heavy pigs. Genetic correlations were estimated in bivariate Bayesian analyses including one BT trait and one production or ham quality trait at a time. Heritability of androstenone, skatole, and indole was 0.41, 0.49, and 0.37, respectively. A moderate negative correlation between skatole and carcass yield (−0.40), and between all BT compounds and backfat (from −0.26 to −0.55) was observed. Conversely, positive correlations (from 0.11 to 0.54) were found between skatole and ham fat thickness traits. Correlations between BT compounds and iodine number ranged from −0.07 (for androstenone) to −0.64 (for skatole), whereas those with PUFA ranged from −0.13 (for indole) to −0.33 (for skatole). Hence, reducing BT could decrease ham fat thickness and increase unsaturated fatty acids, with potential negative impacts on product quality. Full article

This trial evaluated the effect of sex on the blood, growth, carcass, meat quality, and boar taint compounds in male Alentejano (AL) pigs (n = 30). From ~40 to 130 kg LW, castrated (C) and intact pigs (I and IExp groups) were fed commercial diets ad libitum. Between ~130 and 160 kg (slaughter), C and I pigs continued on commercial diets, while IExp were fed an experimental diet containing locally produced pulses and by-products aimed at reducing boar taint. At ~160 kg, blood urea levels were higher in IExp than C pigs, triacylglycerols were lower in both intact groups, and cortisol was lower in IExp. IExp pigs exhibited faster growth, improved feed conversion ratio, carcass higher commercial yield and leaner meat than C pigs. The loin intramuscular fat in intact pigs was lower, less saturated and more polyunsaturated, while total collagen was higher. Fat androstenone content was higher in intact pigs and skatole content was similar across treatments, although they were below threshold values for consumer detection. Finally, although boar taint compounds were low in intact AL pigs raised outdoors, adding pulses and by-products to the experimental diet did not result in a reduction in fat skatole content compared to pigs fed the commercial diet. Full article

Immunocastration and rearing of entire males (EMs) are sustainable alternatives to surgical castration. However, these animal carcasses have variable risk of boar taint and should be identified at the slaughter line. We aimed to identify a simple and reliable indicator of androstenone-related boar taint by evaluating pelvic urogenital tract weight as a marker of boar-taint animals at the slaughter line. The pelvic urogenital tract, testes, and accessory sex glands of EMs and immunocastrates (ICs) were collected, dissected, and weighed, before colorimetric measurements of testicular tissue. Additionally, GnRH antibody titers and testosterone, androstenone, and skatole levels were determined. Our results showed that 81.8% of EMs had androstenone levels above the risk threshold (>0.5 µg/g fat; EM/A^high subgroup), whereas in ICs, the C/A^high subgroup with androstenone >0.5 µg/g fat accounted for only 4.3%. Androstenone levels correlated negatively with GnRH antibody titers and positively with testosterone levels and reproductive organ weights. Identification of ICs with androstenone levels above the threshold (IC/A^high subgroup) may be achieved via testes or pelvic urogenital tract weight measurements. However, in EMs, the latter is a more reliable parameter. A principal component analysis based on these variables and hierarchical clustering also distinguished the A^high from the A^low subgroup, irrespective of IC/EM. The findings highlight the possible use of pelvic urogenital tract weight along with testes weight as a simple, reliable, and efficient morphometric indicator for identifying androstenone-positive carcasses of different sex categories. Full article

Reduced-fat cured sausages were evaluated as a strategy to reduce boar taint in entire male pork products with high levels of androstenone and skatole, both lipophilic. Three fuet-type sausages (two replicates each) were developed: the control (C) (60% lean, 33.69% fat), and two reduced-fat (60% lean 21.19% fat) products; R1, 6% inulin, 0.5% β-glucan and R2, 3% inulin, 0.5% grape skin and 1% β-glucan. All of them were prepared from entire male pork with an androstenone concentration of 6.887 µg/g and 0.520 µg/g of skatole. Significant differences (p ≤ 0.001) in the moisture content were described between the fuet R1 and the C and R2, which obtained the highest percentage. Regarding the CIELAB, the C samples had the highest L* value, while the R2 sausages were the darkest. Boar taint was reduced in both R1 and R2, with a greater reduction in R2 (p ≤ 0.000). The addition of inulin and β-glucan in fuet R1 provided a similar technological and sensory profile to C. However, both strategies provided a reduction of sexual odour, which was higher when grape skins were included. In addition, R2 gave a characteristic sausage with more odour and flavour, dark colour and overall rating than C and R1. Full article

Boar taint is an unpleasant odour and flavour released during heat treatment of pork from uncastrated male pigs. The two main compounds responsible for boar taint are androstenone and skatole. Androstenone is a steroid hormone formed in the testis during sexual maturity. Skatole is a product of microbial degradation of the amino acid tryptophan in the hindgut of pigs. Both of these compounds are lipophilic, which means that they can be deposited in adipose tissue. Several studies have reported heritability estimates for their deposition from medium (skatole) to high magnitudes (androstenone). In addition to efforts to influence boar taint through genetic selection, much attention has also been paid to reducing its incidence using various feeding strategies. From this point of view, research has focused especially on the reduction in skatole content by supplementation of feed additives into the nutrition of entire male pigs. Promising results have been achieved using hydrolysable tannins in the diet. To date, most studies have investigated the effects of tannins on the production and accumulation of skatole in adipose tissue, intestinal microbiota, growth rate, carcasses and pork quality. Thus, the objective of this study was, in addition to determining the effects of tannins on androstenone and skatole accumulation, to assess the effects of tannins on the sensory traits of meat from entire males. The experiment was performed on 80 young boars—progeny of several hybrid sire lines. Animals were randomly assigned to one control and four experimental groups (each numbering 16). The control group (T0) received a standard diet without any tannin supplementation. Experimental groups were supplemented with 1% (T1), 2% (T2), 3% (T3) or 4% (T4) SCWE (sweet chestnut wood extract) rich in hydrolysable tannins (Farmatan). Pigs received this supplement for 40 days prior to slaughter. Subsequently, the pigs were slaughtered, and sensory analysis was applied to evaluate the odour, flavour, tenderness and juiciness of the pork. The results showed a significant effect of tannins on skatole accumulation in adipose tissue (p = 0.052–0.055). The odour and flavour of the pork were not affected by tannins. However, juiciness and tenderness were reduced by higher tannin supplementation (T3–T4) compared to the controls (p < 0.05), but these results were sex-dependent (in favour of men compared to women). Generally, women rated tenderness and juiciness worse than men regardless of the type of diet. Full article

The heterogeneity of Italian manufacturing processes results in the production of a large variety of pork products. In Sardinia, boars are raised and butchered to produce charcuterie. These animals are castrated before slaughter as androstenone would otherwise taint the meat, rendering it unfit for human consumption. However, to date, the literature concerning surgical orchiectomy in adult boars is limited. The goal of this study is to assess whether a sutureless swine orchiectomy procedure is feasible. Additionally, this study aims to determine the appropriate traction force needed to tie knots in the deferens duct of pigs of different weights and ages. Two groups were created: the first (n = 91) underwent orchiectomy by suture ligation; the second (n = 20) was castrated using the sutureless technique. Deferens ducts of animals in the first group (n = 182) were collected following castration, and their tensile strength was measured. Pearson’s linear correlation was used to determine the relationship between the maximum traction force and weight and age groups. A correlation of 0.99 and 0.96 was shown between traction force and age and traction force and weight, respectively. In accordance with these results, sutureless castration was performed on 20 animals, calibrating the pulling force needed according to the age and weight of the boars. No complications were observed during the feasibility study, thus validating sutureless orchiectomy in adult boars. Full article

We discovered odorous 16-androstenes (Androstenone and Androstenol) in endangered mouse deer during a captive breeding program. This study examined the molecular characteristics, their synthesis pathway, and the possible functional role of these compounds in the reproduction of mouse deer. CYP17A1 and CYB5 genes were cloned and expressed in HEK-293, COS-7 cell lines, and gonads of mouse deer to investigate the CYP17A1 gene’s andien-β-synthase activity towards the synthesis of 16-androstenes in mouse deer. An enzyme immunoassay was further developed and standardized to measure fecal androstenone during the reproductive cycles of mouse deer. Results showed that the mouse deer CYP17A1 gene possesses andien-β-synthase activity and could transform pregnenolone into 5,16-androstadien-3β-ol. The expression of the CYP17A1 gene upregulated in the testis and ovary compared to other tissues in mouse deer. Significantly elevated androstenone and estrogens were recorded prior to delivery and postpartum estrus/mating in mouse deer. Further, there were weak correlations between fecal androstenone and estrogens/androgens in mouse deer during the breeding season. These findings suggest that androstenone probably plays a role in the reproductive activities of mouse deer. This knowledge can be used for captive breeding programs of mouse deer in India and elsewhere. Full article