2-Oxidation, 3-methyl hydroxylation, and 6-hydroxylation of skatole, a contributor to the odour of boar-tainted pork meat, mediated by porcine liver microsomal cytochromes P450 1A2, 2A19, 2E1, and 3A22.

The 2-oxidation, 3-methyl hydroxylation, and 6-hydroxylation of skatole (a contributor to boar taint) mediated by minipig liver microsomes and recombinant P450 enzymes expressed in bacterial membranes were investigated.At low substrate concentrations of 10 µM, the formation rates of indole-3-carbinol, 6-hydroxyskatole, and the sum of 3-methyloxindole, indole-3-carbinol, and 6-hydroxyskatole in male minipig liver microsomes were significantly lower than those in female minipig liver microsomes.Compensatory 3-methyloxindole and indole-3-carbinol formation in minipig liver microsomes, which lack 6-hydroxyskatole formation in males, was mediated partly by liver microsomal P450 1A2 and P450 1A2/2E1, respectively. These enzymes were suppressed by typical P450 inhibitors in female minipig liver microsomes.Among the 14 pig P450 forms evaluated, P450 2A19 was the dominant form mediating 3-methyloxindole, indole-3-carbinol, and 6-hydroxyskatole formation from skatole at substrate concentrations of 100 µM. Positive cooperativity was observed in 3-methyloxindole formation from skatole mediated by male minipig liver microsomes and by pig P450 3A22 with Hill coefficients of 1.2-1.5.These results suggest high skatole 2-oxidation, 3-methyl hydroxylation, and 6-hydroxylation activities of pig P450 2A19 and compensatory skatole oxidations mediated by pig P450 1A2, 2E1, or 3A22 in male minipig liver microsomes.

Development of an analytical method for the determination of sterol compounds in boars' saliva.

"Boar taint" compounds influence the sexual behavioral responses of sows and stimulate their reproduction. This paper reports a fast, easier, and a non-invasive analytical method for the analysis of three "boar taint" compounds in boar' saliva samples: androstenone, androsten-3α-ol, and androsten-3ß-ol. This method was developed and validated based on solid-phase extraction (SPE) and multidimensional gas chromatography-mass spectrometry (MDGC-MS). All the compounds were detected without derivatization. This method affords good reproducibility (4%-8%), accuracy (80%-105%), precision (5.5%-9.1%), linearity (R2 = 0.98-0.99), and lower limits of quantitation (LLOQ) (0.1-0.2 µg/L). Although the presence of these compounds in saliva has been known for a long time, no simple and easy analytical method has been developed.

Genomic background and genetic relationships between boar taint and fertility traits in German Landrace and Large White.

BACKGROUND: Due to ethical reasons, surgical castration of young male piglets in their first week of life without anesthesia will be banned in Germany from 2021. Breeding against boar taint is already implemented in sire breeds of breeding organizations but in recent years a low demand made this trait economically less important. The objective of this study was to estimate heritabilities and genetic relationships between boar taint compounds androstenone and skatole and maternal/paternal reproduction traits in 4'924 Landrace (LR) and 4'299 Large White (LW) animals from nucleus populations. Additionally, genome wide association analysis (GWAS) was performed per trait and breed to detect SNP marker with possible pleiotropic effects that are associated with boar taint and fertility. RESULTS: Estimated heritabilities (h2) were 0.48 (±0.08) for LR (0.39 ± 0.07 for LW) for androstenone and 0.52 (±0.08) for LR (0.32 ± 0.07 for LW) for skatole. Heritabilities for reproduction did not differ between breeds except age at first insemination (LR: h2 = 0.27 (±0.05), LW: h2 = 0.34 (±0.05)). Estimates of genetic correlation (rg) between boar taint and fertility were different in LR and LW breeds. In LR an unfavorable rg of 0.31 (±0.15) was observed between androstenone and number of piglets born alive, whereas this rg in LW (- 0.15 (±0.16)) had an opposite sign. A similar breed-specific difference is observed between skatole and sperm count. Within LR, the rg of 0.08 (±0.13) indicates no relationship between the traits, whereas the rg of - 0.37 (±0.14) in LW points to an unfavorable relationship. In LR GWAS identified QTL regions on SSC5 (21.1-22.3 Mb) for androstenone and on SSC6 (5.5-7.5 Mb) and SSC14 (141.1-141.6 Mb) for skatole. For LW, one marker was found on SSC17 at 48.1 Mb for androstenone and one QTL on SSC14 between 140.5 Mb and 141.6 Mb for skatole. CONCLUSION: Knowledge about such genetic correlations could help to balance conventional breeding programs with boar taint in maternal breeds. QTL regions with unfavorable pleiotropic effects on boar taint and fertility could have deleterious consequences in genomic selection programs. Constraining the weighting of these QTL in the genomic selection formulae may be a useful strategy to avoid physiological imbalances.

Characterization of eQTLs associated with androstenone by RNA sequencing in porcine testis.

Characterization of genetic variants affecting genome-wide gene expression levels (expression quantitative trait loci or eQTLs) in pig testes may improve our understanding of genetic architecture of boar taint (an animal welfare trait) and helps in genome-assisted or genomic selection programs. The aims of this study were to identify eQTLs associated with androstenone, to find candidate eQTLs for low androstenone, and to validate the top eQTL by reverse transcriptase quantitative PCR (RT-qPCR). Gene expression profiles were obtained by RNA sequencing in testis from Danish cross-bred pigs and genotype data by 80K single nucleotide polymorphism panel. A total of 262 eQTLs [false discovery rate (FDR) < 0.05] were identified by using two software packages: Matrix eQTL and Krux eQTL. Of these, 149 cis-acting eQTLs were significantly associated with androstenone concentrations and gene expression (FDR < 0.05). The eQTLs were associated with several genes of boar taint relevance including CYP1A2, CYB5D1, and SPHK2. One eQTL gene, AMPH, was differentially expressed (FDR < 0.05) and affected by chicory. Five candidate eQTLs associated with low androstenone concentrations were discovered, including the top eQTL associated with CYP1A2. RT-qPCR confirmed target gene expression to be significantly (P < 0.05) different based on eQTL genotypes. Furthermore, eQTLs were enriched as QTLs for 15 boar taint related traits from the PigQTLdb. This is the first study to report eQTLs in testes of commercial crossbred pigs used in pork production and to reveal genetic architecture of boar taint. Potential applications include development of a DNA test and in advanced genomic selection models for boar taint.

The Mode of Action of Chicory Roots on Skatole Production in Entire Male Pigs Is neither via Reducing the Population of Skatole-Producing Bacteria nor via Increased Butyrate Production in the Hindgut.

The effect of high levels of dietary chicory roots (25%) and intracecal exogenous butyrate infusion on skatole formation and gut microbiota was investigated in order to clarify the mechanisms underlying the known reducing effect of chicory roots on skatole production in entire male pigs. A Latin square design with 3 treatments (control, chicory, and butyrate), 3 periods, and 6 animals was carried out. Chicory roots showed the lowest numerical levels of skatole in both feces and plasma and butyrate infusion the highest. In the chicory group, an increased abundance of the skatole-producing bacterium Olsenella scatoligenes compared to the control group (P = 0.06), and a numerically higher relative abundance of Olsenella than for the control and butyrate groups, was observed. Regarding butyrate-producing bacteria, the chicory group had lower abundance of Roseburia but a numerically higher abundance of Megasphaera than the control group. Lower species richness was found in the chicory group than in the butyrate group. Moreover, beta diversity revealed that the chicory group formed a distinct cluster, whereas the control and butyrate groups clustered more closely to each other. The current data indicated that the skatole-reducing effect of chicory roots is neither via inhibition of cell apoptosis by butyrate nor via suppression of skatole-producing bacteria in the pig hindgut. Thus, the mode of action is most likely through increased microbial activity with a corresponding high incorporation of amino acids into bacterial biomass, and thereby suppressed conversion of tryptophan into skatole, as indicated in the literature.IMPORTANCE Castration is practiced to avoid the development of boar taint, which negatively affects the taste and odor of pork, and undesirable aggressive behavior. Due to animal welfare issues, alternatives to surgical castration are sought, though. Boar taint is a result of high concentrations of skatole and androstenone in back fat. Skatole is produced by microbial fermentation in the large intestine, and therefore, its production can be influenced by manipulation of the microbiota. Highly fermentable dietary fiber reduces skatole production. However, various theories have been proposed to explain the mode of action. In order to search for other alternatives, more efficient or less expensive, to reduce skatole via feeding, it is important to elucidate the mechanism behind the observed effect of highly fermentable dietary fiber on skatole. Our results indicate that highly fermentable dietary fiber does not affect skatole production by reducing the number of skatole-producing bacteria or stimulating butyrate production in the large intestine.

Meat quality of pork loins from Hereford×Berkshire female and intact male pigs reared in an alternative production system.

Objective: The objective of the present study was to investigate pork quality from Hereford×Berkshire (HB) female and intact male pigs reared outdoors in an alternative production system. Methods: Berkshire purebred sows were artificially inseminated, once in the fall and again in the spring of the following year, with semen from Hereford boars and managed free of antibiotics in an outdoor hoop structure until the last month of pregnancy, after which they were moved to a pasture-based unit of 0.8 hectares with individual lots with a farrowing hut, shade, and water ad libitum. Piglets were weaned at 4 weeks of age and housed in a deep-bedded hoop structure, grouped by sex. Animals were harvested at market weight of 125 kg, approximately 200 days of age. Hot carcass weight was collected at the time of the harvest. After 24 hours of refrigeration, carcass characteristics were measured. Longissimus dorsi samples collected from the right side loin. Loins were cut into 2.54-cm thick chops and were used to measure marbling score, color score, drip loss, and ultimate pH. Sensory panel tests were conducted as well at North Carolina State University. For pork characteristics and sensory panel data, trial and sex were included in the statistical model as fixed effects. Hot carcass weight was included in the model as a covariate for backfat thickness. Results: Neither the subjective nor the objective color scores displayed any differences between the boars and the gilts. No difference was found for pH and marbling score between trials or sexes. Gilts had a thicker backfat measurement at the last lumbar and a narrower longissimus muscle area measurement when compared to the boars. The only difference in the sensory characteristics was found between the trials for texture and moisture scores. Conclusion: Consumers were not able to detect boar taint under the condition of this study, which is that the intact males were reared outdoors. Additional trials would be necessary; however, based on the results of the present study, outdoor rearing can be suggested as a solution to the issue of boar taint.

Reduction of androstenone perception in pan-fried boar meat by different masking strategies.

BACKGROUND: Consumers highly sensitive to androstenone María (AND) will probably reject meat from entire male pigs, which tends to have high levels of this hormone. To avoid this, the effect of different masking strategies (sprinkling with mixed spices or fennel, marinating and breading with garlic-parsley or curry) on the sensory parameters of pork loin chops obtained from entire animals with high levels of AND (1.0-2.9 mg kg-1 AND in fat) and castrated animals (<0.4 mg kg-1 AND in fat), both with low levels of skatole (<0.1 mg kg-1 skatole in fat) was investigated. RESULTS: The garlic-parsley breadcrumbs led to the highest reduction in the perception of AND compared with the other masking strategies used, and preserved the juiciness of the product. There was a negative correlation between AND and fat content. CONCLUSION: AND odor and flavor can be reduced in meat from entire male pigs by using suitable strategies, the best strategy being the garlic-parsley breadcrumbs. © 2017 Society of Chemical Industry.

Lipolysis and aroma generation as mechanisms involved in masking boar taint in sodium reduced fermented sausages inoculated with Debaryomyces hansenii yeast.

BACKGROUND: The use of boar back fat for processing of fermented sausages may cause the presence of abnormal odours. In dry-cured products, ripening time is essential to develop the sensory characteristics. Yeast has been proposed as an alternative to mask boar taint odour through its metabolic activity but it is necessary to elucidate which mechanisms are involved. The aim is to study the effect of Debaryomyces hansenii inoculation on the lipolysis process and generation of aroma compounds in fermented sausages manufactured with boar back fat at two different ripening times. RESULTS: D. hansenii inoculated sausages had a higher degree of lipolysis as demonstrated by higher content of free fatty acids, ester compounds and branched aldehydes which contribute the fruity odour. The increase in lipolysis produced by D. hansenii inoculation was not followed by an increase in oxidation during processing possibly due to the metabolic activity of yeast. The effect of back fat type was scarcely appreciated whereas ripening time had a stronger effect on sausage. Boar sausages were characterised by a lower polyunsaturated fatty acid profile and lesser lipolysis than gilt sausages. CONCLUSION: Yeast inoculation with D. hansenii and long ripening time were appropriate strategies to limit the perception of boar taint in dry fermented sausages. © 2017 Society of Chemical Industry.

Dietary approaches reducing boar taint-Importance of Lawsonia intracellularis colonisation for interpreting results.

In the fattening of male pigs, boar odour is a major problem with regard to the acceptance of the meat by consumers. Skatole can be one cause. Tryptophan from non-digested feed ingredients and intestinal cell debris can be the precursor in skatole formation. Lawsonia intracellularis, one of the most widespread pathogens in swine, promotes the epithelial cell turnover and might favour the tryptophan influx into the hindgut. Therefore, the question arises how far the severity of a Lawsonia intracellularis infection has an effect on results of dietary experiments with specific issues. Fifty finishing boars from a specific pathogen-free farm were randomly allotted to ten boxes in five feeding groups. Natural developing Lawsonia intracellularis colonisation was monitored serologically (twice individually) and molecular biologically (weekly individually). Over 4 weeks, animals were fed either a finely ground pelleted diet (FP), a coarsely ground meal diet (CM), a meal diet either with 22% cracked corn (CORN), 16.9% dried whey (WHEY) or 30% raw potato starch (RPS). Fifty % of animals showing lower differences in serological Lawsonia intracellularis values between the start and the end of the trial were characterised by a higher dry matter content in faeces (256 ± 29.4 vs. 239 ± 23.6 g/kg). Lawsonia intracellularis-negative caecal samples showed the highest butyrate concentrations (27.2 ± 7.53 mmol/kg). Lawsonia intracellularis-negative faecal samples of group FP showed the highest DM levels in faeces (neg: 290 ± 46.1/pos: 250 ± 52.2 g/kg); negative samples from group RPS had the lowest values (217 ± 24.4 g/kg). Lawsonia intracellularis-negative faecal samples from the group CM were lower in skatole than positive samples (82.8 ± 32.8 vs. 119 ± 29.3 µg/g DM). RPS group samples without pathogen detection had the lowest skatole concentrations (30.5 ± 36.3 µg/g DM). This study provides first evidence that clinically unremarkable colonisation with intestinal pathogens might influence the results of dietary approaches.

Fine mapping of a QTL affecting levels of skatole on pig chromosome 7.

BACKGROUND: Previous studies in the Norwegian pig breeds Landrace and Duroc have revealed a QTL for levels of skatole located in the region 74.7-80.5 Mb on SSC7. Skatole is one of the main components causing boar taint, which gives an undesirable smell and taste to the pig meat when heated. Surgical castration of boars is a common practice to reduce the risk of boar taint, however, a selection for boars genetically predisposed for low levels of taint would help eliminating the need for castration and be advantageous for both economic and welfare reasons. In order to identify the causal mutation(s) for the QTL and/or identify genetic markers for selection purposes we performed a fine mapping of the SSC7 skatole QTL region. RESULTS: A dense set of markers on SSC7 was obtained by whole genome re-sequencing of 24 Norwegian Landrace and 23 Duroc boars. Subsets of 126 and 157 SNPs were used for association analyses in Landrace and Duroc, respectively. Significant single markers associated with skatole spanned a large 4.4 Mb region from 75.9-80.3 Mb in Landrace, with the highest test scores found in a region between the genes NOVA1 and TGM1 (p < 0.001). The same QTL was obtained in Duroc and, although less significant, with associated SNPs spanning a 1.2 Mb region from 78.9-80.1 Mb (p < 0.01). The highest test scores in Duroc were found in genes of the granzyme family (GZMB and GZMH-like) and STXBP6. Haplotypes associated with levels of skatole were identified in Landrace but not in Duroc, and a haplotype block was found to explain 2.3% of the phenotypic variation for skatole. The SNPs in this region were not associated with levels of sex steroids. CONCLUSIONS: Fine mapping of a QTL for skatole on SSC7 confirmed associations of this region with skatole levels in pigs. The QTL region was narrowed down to 4.4 Mb in Landrace and haplotypes explaining 2.3% of the phenotypic variance for skatole levels were identified. Results confirmed that sex steroids are not affected by this QTL region, making these markers attractive for selection against boar taint.

Exploiting whole genome sequence data to fine map and characterize candidate genes within a quantitative trait loci region affecting androstenone on porcine chromosome 5.

Male piglets are routinely castrated to eliminate boar taint. However, this treatment is undesirable, and alternative approaches, including genetic strategies to reduce boar taint, are demanded. Androstenone is one of the causative agents of boar taint, and a QTL region affecting this pheromone has previously been reported on SSC5: 22.6-24.8 Mb in Duroc. The QTL region is one of the few reported for androstenone that does not simultaneously affect levels of other sex steroids. The main objective of this study was to fine map this QTL. Whole genome sequence data from 23 Norwegian Duroc boars were analyzed to detect new polymorphisms within the QTL region. A subset of 161 SNPs was genotyped in 834 Duroc sires and analyzed for association with androstenone in adipose tissue and testosterone, estrone sulphate and 17ß-estradiol in blood plasma. Our results revealed 100 SNPs significantly associated with androstenone levels in fat (P < 0.001) with 94 of the SNPs being in strong linkage disequilibrium in the region 23.03-24.27 Mb. This haplotype block contains at least four positional candidate genes (HSD17B6, SDR9C7, RDH16 and STAT6) involved in androstenone biosynthesis. No significant associations were found between any of the SNPs and levels of testosterone and estrogens, confirming previous findings. The amount of phenotypic variance explained by single SNPs within the haplotype block was as high as 5.4%. As the SNPs in this region significantly affect levels of androstenone without affecting levels of other sex steroids, they are especially interesting as genetic markers for selection against boar taint.

Development of recombinant antibody-based enzyme-linked immunosorbent assay (ELISA) for the detection of skatole.

The occurrence of boar taint and the European Commission recommendation to discontinue the surgical castration of pigs by the year 2018 creates an urgent need for new analytical methods that are simple, affordable, and suitable for field testing. We describe the generation and engineering of a skatole-specific antibody derived from a synthetic antibody library and the development of ELISA for its detection. The immunoassay is capable of detecting skatole with IC50 of 222 µg L(-1), which is within the analytical threshold level suggested for skatole, and with low cross-reactivity interference from other indolic compounds.

Simultaneous quantification of the boar-taint compounds skatole and androstenone by surface-enhanced Raman scattering (SERS) and multivariate data analysis.

This study investigates the feasibility of using surface-enhanced Raman scattering (SERS) for the quantification of absolute levels of the boar-taint compounds skatole and androstenone in porcine fat. By investigation of different types of nanoparticles, pH and aggregating agents, an optimized environment that promotes SERS of the analytes was developed and tested with different multivariate spectral pre-processing techniques, and this was combined with variable selection on a series of analytical standards. The resulting method exhibited prediction errors (root mean square error of cross validation, RMSECV) of 2.4 × 10(-6) M skatole and 1.2 × 10(-7) M androstenone, with a limit of detection corresponding to approximately 2.1 × 10(-11) M for skatole and approximately 1.8 × 10(-10) for androstenone. The method was subsequently tested on porcine fat extract, leading to prediction errors (RMSECV) of 0.17 µg/g for skatole and 1.5 µg/g for androstenone. It is clear that this optimized SERS method, when combined with multivariate analysis, shows great potential for optimization into an on-line application, which will be the first of its kind, and opens up possibilities for simultaneous detection of other meat-quality metabolites or pathogen markers. Graphical abstract Artistic rendering of a laser-illuminated gold colloid sphere with skatole and androstenone adsorbed on the surface.

Differential expression of CYB5A in Chinese and European pig breeds due to genetic variations in the promoter region.

Cytochrome b5 (CYB5A) is an important electron transfer protein with homologues in a number of different organisms. In pigs, CYB5A is related to boar taint because of its role in androstenone biosynthesis. To determine the variety of CYB5A expression in pig breeds, genetic variations in the porcine CYB5A promoter region in both Chinese and European pig breeds were examined. Three single nucleotide polymorphisms (NC_010443.4:g.165901487delG, g.165901767T>C and g.165902078C>T) were identified in the porcine CYB5A promoter region. These SNPs occurred in different frequencies in Chinese and European pigs. Chinese pigs were primarily haplotype B (denoted as delG-C-T: the position of nt 165901487 of the CYB5 gene is a G deletion, nt 165901767 is C and nt 165902078 is T), except for Licha black pigs, which were primarily haplotype A (denoted as G-T-C: nt 165901487 is G, nt 165901767 is T and nt 165902078 is C), similar to European pigs. Quantitative PCR data from liver tissues demonstrated that haplotype B individuals had higher CYB5A expression than did those with haplotype A. This was confirmed by in vitro cell transfection assays, in which haplotype B individuals had higher reporter activity than did those with haplotype A. In silico analysis predicted that Myc-associated zinc-finger protein (MAZ) is a potential transcription factor at position 165901767. Electrophoretic mobility shift assays showed this polymorphism affects the stable binding of transcription factors to the CYB5A promoter, which in turn affects the expression levels of this gene. Therefore, this variation of the porcine CYB5A promoter region may explain the differences in androstenone accumulation between Chinese and European pig breeds and may also prove useful as a genetic marker to distinguish the origin of different pig breeds.

Asian low-androstenone haplotype on pig chromosome 6 does not unfavorably affect production and reproduction traits.

European pigs that carry Asian haplotypes of a 1.94-Mbp region on pig chromosome 6 have lower levels of androstenone, one of the two main compounds causing boar taint. The objective of our study was to examine potential pleiotropic effects of the Asian low-androstenone haplotypes. A single nucleotide polymorphism marker, rs81308021, distinguishes the Asian from European haplotypes and was used to investigate possible associations of androstenone with production and reproduction traits. Eight traits were available from three European commercial breeds. For the two sow lines studied, a favorable effect on number of teats was detected for the low-androstenone haplotype. In one of these sow lines, a favorable effect on number of spermatozoa per ejaculation was detected for the low-androstenone haplotype. No unfavorable pleiotropic effects were found, which suggests that selection for low-androstenone haplotypes within the 1.94 Mbp would not unfavorably affect the other eight relevant traits.

Hepatic Gene Expression and Metabolite Profiles of Androstenone and Skatole Relative to Plasma Estrone Sulfate Levels in Boars.

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.

Enhanced HPLC Method for Boar Taint Quantification.

Boar taint is an unpleasant odour found in the carcasses of entire male pigs, resulting from androstenone and skatole accumulation during pubertal development, and impacting pork quality. This study proposes the validation of an adapted chromatographic method for quantifying skatole and androstenone in the pigs' liquid fat using fluorescence detection. A good chromatographic separation was achieved, with skatole (SKA) and androstenone (AND) elution at 4.4 and 9.9 min., respectively. An external calibration method was applied, with calibration curves correlation coefficient of 0.9999 for both analytes. Detection limit values were 1.53 and 16.02 ng/g for SKA and AND, respectively. SKA recovery was 99.72±2.34 % (2.34 % RSD) and 102.84±1.62 % (1.57 % RSD) for AND. Results showed good precision values (repeatability <2.46 % RSD for SKA, <6.85 % RSD for AND; intermediate precision <2.87 % RSD for SKA, <6.98 % RSD for AND). The method's robustness was tested and the values were within the reference ranges. The validation results proved that the adaptation of an existing method resulted in good assessments of robustness, reliability and accuracy.

Feeding strategy in organic pig farming as a lever to improve various quality dimensions of pork.

Since 2022, European specifications for organic pig farming have evolved to distribute 100 % organic feed and to reinforce the link to the soil with feed resources that should primarily be obtained from the farm or the same region. Feeding strategy acts as a lever to improve various quality dimensions of organic (as well as conventional) pork, including intrinsic dimensions (carcass composition, nutritional, organoleptic, technological, sanitary qualities) and extrinsic dimensions related to animal farming (image). Diet may also influence the risk of undesirable odours or flavours that may be found in pork from non-castrated male pigs. This study aimed at evaluating the effects of a specific feeding strategy on several quality dimensions of organic meat from non-castrated male pigs. The experiment was conducted with 77 organic non-castrated male pigs (Piétrain NN × Large White) reared according to organic specifications and distributed in two batches. Within litters, male littermates were allocated at around 33 kg of live weight to either a Control group which received a Control feed (C) corresponding to the organic specifications or in a Bio+ group which received an organic test feed based mainly on French raw materials and which contained more fibres (faba bean and access to forages) and omega-3 fatty acids (linseed, camelina). All pigs were reared in the same building on deep straw bedding (1.3 m2/pig) with free outdoor access (1.0 m2/pig) using one pen per experimental group. Pigs were fed ad libitum until slaughter at about 125 kg live weight. Average daily gain, carcass weight and lean meat content did not differ significantly between C and Bio+ pigs. Compared to C, Bio+ pigs had higher (P < 0.05) ultimate pH in the loin (Longissimus muscle) and ham (Gluteus medius, and Semimembranosus) muscles, associated with a lower Longissimus muscle glycolytic potential (P < 0.001). Loin and ham meat from Bio+ vs. C pigs was lighter (P < 0.05) and had a more intense red colour (P < 0.10). The Bio+ strategy led to a lower Longissimus muscle n-6:n-3 fatty acid ratio (P < 0.001), indicating an improvement in pork nutritional value. Backfat skatole concentration was lower in Bio+ than in C pigs whereas backfat androstenone was higher in Bio+ than in C pigs (P < 0.05). Altogether, we demonstrated that the Bio+ strategy had positive impacts on several qualities of organic pork from non-castrated male pigs.

The results of meat quality traits and sensory characteristics according to the concentration of androstenone in uncastrated pigs.

Pork quality is determined by several attributes, among which odor and taste are the utmost significant. Therefore, this study was aimed to assess the effects of boar odor hormone concentration on the quality traits and sensory acceptability of pork. A total twenty-six (26) non-castrated 3-way crossbred (Landrace × Yorkshire × Duroc) pigs were selected with an average body weight (ABW) 115.6 kg before to slaughter. The three treatment groups (low, medium and high) were divided according to the androstenone concentration. In experiment 1, for meat quality traits carcass was selected based on androstenone concentration: low (LC, 0.64-0.69 µg/g, n = 9), medium (MC, 0.70-0.99 µg/g, n = 7) and high (HC, 1.00-1.69 µg/g, n = 10). In experiment 2, for sensory evaluation carcasses were also selected based on the abovementioned conditions. Results revealed that androstenone concentration not effect on proximate components, meat quality traits and fatty acids except palmitoleic acid. Sensory evaluation data showed that boar taint and meat boar taint were significantly increased in a concentration-dependent manner from low to high, whereas, gravy and meat flavor preference were significantly increased in LC group than HC group. In addition, correlation analysis showed that boar taint and meat boar taint were positively, and gravy and meat flavor preference were negatively correlated with boar taint hormones. In essence, our findings indicate that androstenone concentration had no effect on meat qualities, but a high concentration of androstenone had a negative effect on the sensory characteristics in uncastrated pigs.

Development and validation of a classification model for boar taint detection in pork fat samples.

This study aims to characterize a complete volatile organic compound profile of pork neck fat for boar taint prediction. The objectives are to identify specific compounds related to boar taint and to develop a classification model. In addition to the well-known androstenone, skatole and indole, 10 other features were found to be discriminant according to untargeted volatolomic analyses were conducted on 129 samples using HS-SPME-GC×GC-TOFMS. To select the odor-positive samples among the 129 analyzed, the selection was made by combining human nose evaluations with the skatole and androstenone concentrations determined using UHPLC-MS/MS. A comparison of the data of the two populations was performed and a statistical model analysis was built on 70 samples out of the total of 129 samples fully positive or fully negative through these two orthogonal methods for tainted prediction. Then, the model was applied to the 59 remaining samples. Finally, 7 samples were classified as tainted.