A single nucleotide polymorphism in the CYP2E1 gene promoter affects skatole content in backfat of boars of two commercial Duroc-sired crossbred populations

The prevention of unpleasant boar taint is the main reason for castration of male piglets. This study aimed to investigate how the malodorous compound skatole is affected by a single nucleotide polymorphism (g.2412 C>T at − 586 ATG) in the porcine cytochrome p450 II E1 (CYP2E1) gene. 119 boars of two commercial Duroc-sired crossbred populations raised at different farms were investigated. Skatole and androstenone in backfat averaged 114 ± 125 ng/g and 1206 ± 895 ng/g melted fat, respectively. The frequency of the genotypes CC, CT, and TT was 25, 52, and 23%, respectively. CC boars had the highest average skatole levels (175 ng/g) compared to CT (92 ng/g) and TT (93 ng/g). Applying suggested sensory threshold levels for skatole (> 150 ng/g) and androstenone (> 2000 ng/g), 30% of the carcasses may be unacceptably tainted while the proportion of tainted carcasses is significantly higher within genotype CC (56.7%) compared to genotypes CT (24.3%) and TT (14.8%). Effective reduction of tainted carcasses appears feasible applying marker assisted selection.     

Different scalding techniques do not affect boar taint

The prevention of unpleasant boar taint is the main reason for castration of male piglets. For animal welfare reasons, castration is announced to be banned in the European Community. This study aimed to investigate whether androstenone, skatole and indole in backfat of boars may be reduced by different scalding technologies. To discriminate ante and post mortem effects, carcasses were sampled before and after scalding in two abattoirs using either horizontal (TANK) or vertical (TUNNEL) scalding. Backfat samples were analysed using gas chromatography (androstenone) and liquid chromatography (skatole, indole). Neither TANK nor TUNNEL scalding did significantly reduce malodorous compounds. Skatole and androstenone in backfat obtained after scalding averaged 112 ± 123 ng/g and 1196 ± 885 ng/g melted fat, respectively; significant differences between abattoirs were observed for skatole. Increased skatole levels were tentatively assigned to longer transport duration. Concluding from recent consumer research and subsequent application of suggested sensory rejection thresholds for androstenone (2000 ng/g) and skatole (150 ng/g), nearly 30% of the carcasses may be unacceptably tainted.     

Quantitative determination of the boar taint compounds androstenone,skatole, indole, 3α-androstenol and 3β-androstenol in wild boars (Sus scrofa) reveals extremely low levels of the tryptophan-related degradation products

The major boar taint compounds androstenone and skatole as well as the minor compounds indole, 3α-androstenol and 3β-androstenol were determined in back fat samples of 23 male wild boars by applying a recently published SIDA–HS-SPME–GC/MS method. The boar pheromones androstenone, 3α-androstenol and 3β-androstenol were found in extraordinary high concentrations, resulting in mean values of 3329 ng/g androstenone, 1273 ng/g 3α-androstenol and 545 ng/g 3β-androstenol. Interestingly, skatole was not detectable in about 50% of the boars and negligibly low in all other samples as expressed by a mean skatole value of only 14 ng/g. Indole was also found in every sample, but again in low concentrations with a mean value of 40 ng/g. Possible factors explaining this remarkably low skatole deposition in wild boars such as intestinal flora and anatomy, dietary composition, housing or genetic predisposition are discussed in this paper.     

The effect of age on distribution of skatole and indole levels in entire male pigs in four breeds: Yorkshire, Landrace, Hampshire and Duroc

High skatole and to a lesser degree indole levels in some entire male pigs result in occurrence of off-flavor in meat, called boar taint. In this study, plasma samples from 117 Yorkshire, 134 Landrace, 184 Hampshire and 75 Duroc entire male pigs collected at different ages were analyzed for skatole and slightly fewer samples for indole concentrations. For each breed, a distinct age-related distribution of plasma skatole levels was observed. Skatole levels were increased at approximately 180-200 days of age, reaching very high levels is some individuals. Afterwards, the levels decreased; in Yorkshire and Landrace at approximately 240-260 and in Hampshire and Duroc at 310-360 days of age. The decrease of skatole levels was also observed in samples collected from 15 Landrace boars at two different ages: 16.8 μg/L (SD=17.36) vs. 5.9 μg/L (SD=3.48, p=0.016) in samples taken at the mean age of 210 and 314 days, respectively. Indole levels showed similar age- and breed-related variations to skatole. At the time of increased skatole concentrations (age from 180 to 360 days, depending on breed) 25.5% of Yorkshire, 31.6% of Landrace, 20.3% of Hampshire and 61.1% of Duroc entire male pigs had skatole levels above 12.6 μg/L of plasma, which corresponds to the threshold level of 0.20 μg/g of fat that is used for selecting tainted carcasses. The results indicate that increased levels of skatole in entire male pigs are related to puberty and taking skatole measurements at that age could be advantageous when considering genetic selection to reduce boar taint level in entire male pigs. Breed differences should be also taken into account.     

Effect of housing system, slaughter weight and slaughter strategy on carcass and meat quality, sex organ development and androstenone and skatole levels in Duroc finished entire male pigs

This study aimed at evaluating the effect of housing system (HS), slaughter weight (SW) and strategy (SS) on carcass a nd meat quality, sexual organ development and boar taint in entire males. Twelve pens of 10 pigs were used (two trials). Half of male pens were allowed visual contact with females (MF) and half with males (MM). Half MM or MF were slaughtered at 105 or 130 kg in trial 1, or penwise or by split marketing in trial 2 at 120 kg. Housing system showed no significant effect on carcass or meat quality. MF presented significantly longer testicles and heavier bulbourethral glands compared to MM. The distribution of androstenone and skatole levels was affected by SW but not by HS or SS, samples with androstenone >1 μg/g of the different groups falling within the range of 16 to 22%. All correlations between androstenone and sex organs were significant. Housing system and slaughter strategy did not reduce the risk of boar tainted carcasses.     

Boar taint detection using parasitoid biosensors

The off-flavor boar taint associated with the substances skatole, androstenone, and possibly indole represents a significant problem in the pig husbandry industry. Boar taint may occur in meat from uncastrated sexually mature male pigs; consumers commonly show a strong aversion to tainted meat. Consequently, there is a need for rapid methods to sort out and remove tainted carcasses at the slaughterline. We tested the ability of wasps, Microplitis croceipes to perceive and learn the 3 boar taint compounds both individually and in combination using classical conditioning paradigms. We also established the effectiveness and reliability of boar taint odor detection when wasps were used as biosensors in a contained system called the "wasp hound" using a cohort of trained wasps. We found that the wasps are able to successfully learn indole, skatole and to also detect them when presented a 1:1:1 mixture of all 3 compounds. This was shown for both a single hand-manipulated wasp bioassay and when using the "wasp hound" detector device. In contrast, the wasps showed a weak conditioned response to androstenone at the concentration tested. The estimated gas phase concentrations that the wasps perceived during training were in the range of 10 ± 0.4 pg/s for skatole and indole, and 2 ± 0.5 pg/s for androstenone. We conclude that use of these wasps as biosensors presents a promising method for boar taint detection and discuss future training paradigms that may improve their responses to compounds such as androstenone. Practical Application: The development of a perceptive, inexpensive, and reliable means of detecting boar taint before the product is presented to sensitive consumers.     

Temporal aspects related to the perception of skatole and androstenone, the major boar odour compounds

Pork fat samples from 50 boars slaughtered at a commercial abattoir were analysed for skatole and androstenone concentrations and grouped within a 3×3 matrix representing low, medium and high levels of the odour compounds. A 10 member sensory panel, screened and trained to recognise and quantify skatole and androstenone odour intensities, was used to verify the human perception of boar odour in these pork fat samples immediately after heating (±65°C) and following a cooling period of ten minutes (±25°C). Principal component analysis revealed that the sensory panel differentiated the pork fat samples first and foremost on the basis of the presence or absence of androstenone and/or skatole odours and secondly on the character of the androstenone or skatole odour. Evidence of adaptation towards the odour of skatole was shown by a decreased sensitivity over replicates. Sensory perception of boar odour seems to have a temporal character which can be explained by differences in volatilisation (involving both odourant release and retention) of skatole and androstenone, odour synergism and possibly differences in the properties of the fat matrix of different samples. The results suggest that factors influencing volatilisation of skatole and androstenone should also be considered when predicting sensory responses of boar odour.     

Predicting the likelihood of developing boar taint: Early physical indicators in entire male pigs

Three potential early-age predictors of which boars are likely to develop boar taint (testes volume, skin lesions and dirtiness) were measured on 102 boars every fortnight from 10 weeks of age until slaughter. These predictors were correlated with the level of boar taint according to the hot iron method and the concentrations of skatole and androstenone as determined by chemical analysis. The chance of no/low boar taint according to the hot iron method decreased with higher testes volume (weeks 22 and 24) and increased with skin lesion score (weeks 12, 16 and 18). For the concentrations of androstenone and skatole, the strongest correlation was found with testes volume in week 12. Skin lesions in week 16 were negatively correlated with skatole levels. Dirtiness was negatively correlated with skatole concentrations (week 18) but positively correlated with androstenone concentrations (weeks 20 and 22). Testes volume has the greatest potential for predicting the likelihood of developing boar taint.     

Predicting the likelihood of developing boar taint: Early physical indicators in entire male pigs

Three potential early-age predictors of which boars are likely to develop boar taint (testes volume, skin lesions and dirtiness) were measured on 102 boars every fortnight from 10weeks of age until slaughter. These predictors were correlated with the level of boar taint according to the hot iron method and the concentrations of skatole and androstenone as determined by chemical analysis. The chance of no/low boar taint according to the hot iron method decreased with higher testes volume (weeks 22 and 24) and increased with skin lesion score (weeks 12, 16 and 18). For the concentrations of androstenone and skatole, the strongest correlation was found with testes volume in week 12. Skin lesions in week 16 were negatively correlated with skatole levels. Dirtiness was negatively correlated with skatole concentrations (week 18) but positively correlated with androstenone concentrations (weeks 20 and 22). Testes volume has the greatest potential for predicting the likelihood of developing boar taint.     

Supercritical Carbon Dioxide Extraction of Androstenone and Skatole from Pork Fat

Boar taint, an unpleasant odor observed in pork from some mature intact male pigs, is attributed primarily to the presence of two compounds in boar fat, androstenone (Sα-androst-16-ene-3-one) and skatole (3-methyl indole). A rapid extraction method for the preparation of fat samples for androstenone/skatole screening assays would lead to more efficient use of boar carcasses for pork production. Supercritical carbon dioxide removed 97 ± 2% of the androstenone (extraction at 40°C; 5 min) and 65 ± 3% of the skatole (extraction at 40°C; 20 min) from 0.5 g of boar backfat. Supercritical CO₂ provides the basis for a fairly rapid extraction method to remove compounds associated with boar taint.     

An international study on the importance of androstenone and skatole for boar taint: IV. Simulation studies on consumer dissatisfaction with entire male pork and the effect of sorting carcasses on the slaughter line, main conclusions and recommendations

An international study has been conducted in order to determine the respective contributions of androstenone and skatole to boar taint and their possible variations according to production systems and consumer populations. The presentation of the study and the main results concerning skatole and androstenone levels and data from sensory evaluation or consumer surveys are reported in companion papers. The present paper summarises the main conclusions of the study and gives tentative recommendations. A simulation study was conducted, based on the skatole and androstenone levels currently observed in European populations of entire male pigs and on the results of the consumer surveys. The first part of the simulation study demonstrated that, overall, 6.5% (odour) and 3.0% (flavour) more consumers were dissatisfied with entire male than with gilt pork. The differences were, however, very variable according to countries. Consumer dissatisfaction for the odour of entire male pork was mostly associated with high skatole levels, while androstenone had little influence on it. On the other hand, androstenone and skatole had similar contributions to the level of dissatisfaction for flavour. From the present study it is not possible to determine clear cut-off levels for androstenone/skatole. The regression equations presented in [Matthews, K. R., Homer, D. B., Punter, P., Béague, M. P., Gispert, M., Siret, F., Leask, H., Fonti i Furnols, M., & Bonneau, M. (2000). An international study on the importance of androstenone, skatole for boar taint: III. Consumer survey in seven European countries. Meat Science, 54, 271-283] provide a basis for decision making. However, due to methodological limitations, the results may underestimate consumer reaction to entire male pork. The second part of the simulation study demonstrated that sorting carcasses on the basis of androstenone/skatole would reduce, but not eliminate, differences in consumer dissatisfaction between entire male and gilt pork. For odour, taking androstenone into account did not improve the efficiency obtained from sorting using skatole only. For flavour, sorting using both compounds was more efficient than sorting using skatole only. Sorting out 15% of the entire males, on the basis of skatole only, would result in a difference in the proportion of dissatisfied consumers of 4.2% (odour) or 2.0% (flavour) between entire male and gilt pork. The results of the last part of the simulation study demonstrated that decreasing skatole in entire male pig populations, to levels as low as 0.10 ppm, would still result in a difference in the proportion of dissatisfied consumers of 3.2% (odour) or 1.6% (flavour). To reduce this difference further, the levels of both compounds would have to be reduced still further. The lowest difference that can be achieved is 2.3% (odour) or 0.4% (flavour). The conclusions of the present study may differ according to whether immediate commercial applications or long-term goals are considered. On the basis of the skatole and androstenone levels currently observed in entire male pig populations, sorting out procedures based on skatole is the easiest way to rapidly achieve a significant decrease in consumer dissatisfaction with entire male pork. In most countries, however, this will not be sufficient to obtain the same level of acceptability as with gilts. In the long term, a sharp reduction in both skatole and androstenone would have to be achieved in entire male pig populations to obtain low differences in acceptability between entire male and gilt pork.     

Boar taint in pigs selected for components of efficient lean growth rate

Genetic and nutritional effects on the boar taint traits of androstenone, skatole and indole and the eating quality traits associated with boar taint were examined by testing animals from four selection lines and a control line on isoenergetic diets, which differed in ileal digestible lysine: digestible energy (0.40, 0.76 and 1.12 g lysine/MJ DE). The selected lines resulted from seven generations of selection for high daily food intake, lean food conversion ratio (LFC) and lean growth rate on ad libitum (LGA) or restricted (LGS) feeding regimes in a Large White population. During performance test, from 30 to 90 kg, boars were fed on either ad libitum or restricted (0.75 g/g ad libitum daily food intake) feeding regimes. A sensory panel assessed heated fat samples for androstenone odour, skatole odour and abnormal odour. There were no significant differences between the selection and control lines or diets for log transformed fat content of androstenone, skatole, indole. The significant diet with feeding regime interaction for log transformed fat content of skatole and indole were essentially due to significantly higher log transformed fat contents with ad libitum feeding of the high lysine diet compared to restricted feeding (skatole: -1.94 vs -3.06, s.e.d. 0.43; indole: -3.44 vs -4.22, s.e.d. 0.28), as differences between feeding regimes on diets A and C were not significantly different from zero. There were no significant differences between selection and control lines for sensory panel score for abnormal odour or androstenone odour, but the LFC and LGA selection lines had a significantly higher skatole odour score than the LGS selection line. Neither diet nor feeding regime had any significant effect on sensory panel assessment of odour. Log transformed fat content of androstenone and skatole were significantly correlated with sensory panel score for skatole odour (0.37 and 0.46, s.e. 0.12), but not with sensory panel score for androstenone odour (0.06 and 0.09), such that they would not be useful predictors of androstenone odour.     

The problem of taint in pork-II. The influence of skatole, androstenone and indole, Presented individually and in combination in a model lipid base, on odour perception

Responses to varying concentrations of androstenone and skatole in a neutral lipid base were studied using a trained 10-member panel. Concentrations for skatole and androstenone ranged between 0-1.32 and 0-6.0 μg g(-1), respectively. Odour, androstenone and skatole intensities were assessed by the line scaling method. Triangular tests showed that the panel significantly distinguished indole from androstenone, p ≤ 0.01, but not from skatole nor mixtures of androstenone and skatole. Regression analyses showed linear relationships between skatole and androstenone concentrations and odour intensity but with a significant (p ≤ 0.01) androstenone-assessor interaction. Correlation coefficients with odour intensity were 0.64 and 0.63 for androstenone and skatole concentrations, respectively. Response surface curves showed that odour intensity depended on the linear terms of skatole, androstenone and on the quadratic term of androstenone. Androstenone and skatole intensity ratings revealed that the effect of skatole was enhanced by the presence of androstenone, but not the reverse. It was concluded that whilst both androstenone and skatole may be important for the sorting (classification) of pork, the importance of androstenone lies in its detection threshold whilst for skatole, the acceptability threshold may be a suitable criterion for quality control purposes.     

Relationships between skatole and androstenone accumulation, and cytochrome P4502E1 expression in Meishan×Large White pigs

The effects of age, castration and diet on accumulation of skatole and androstenone in the backfat of 50% Meishan male pigs, was studied in relation to boar taint. Skatole and androstenone deposition in backfat of entire males was increased with age (114, 144 and 174 days). Castration significantly decreased skatole and androstenone levels in all age groups. The level of cytochrome P4502E1, the initial enzyme involved in hepatic skatole metabolism, was increased by castration at all ages. These results suggest that the decrease in androstenone levels following castration affects the regulation of cytochrome P4502E1 expression, and as a consequence, results in a decrease in skatole levels. The use of sugar beet feed in the diet decreased the level of skatole and increased that of cytochrome P4502E1 expression, but did not affect androstenone levels. It is suggested that skatole accumulation is reduced both by castration and by diet via induction of cytochrome P4502E1. However, the mechanism of induction of cytochrome P4502E1 by diet is different from that involved in its induction by castration.     

Acceptability of entire male pork with various levels of androstenone and skatole by consumers according to their sensitivity to androstenone

Consumer acceptability of entire male pork at eating was assessed in three experiments. The 140 consumers involved in each experiment were classified as insensitive (INSENS) to the odor of pure androstenone or sensitive perceiving it as pleasant (SENS-PLEA) or unpleasant (SENS-UNPL). Entire male pork with very low skatole and androstenone levels (LS-LA) was as well accepted as gilt pork, whatever the consumer category. Entire male pork with elevated levels in both skatole and androstenone (HS-HA) was clearly differentiated from LS-LA pork by SENS-UNPL, but not by SENS-PLEA or INSENS consumers. Whatever the consumer category, entire male pork with elevated levels of androstenone and very low levels of skatole (LS-HA and LS-HHA) were not significantly differentiated from LS-LA pork. The results suggest that, in the conditions of the present experiment, androstenone and skatole totally explain boar taint at eating and that the acceptability threshold for androstenone, in the absence of skatole, is in the range of 2-3 μg/g liquid fat.     

Influence of differences in sensitivity of Spanish and German consumers to perceive androstenone on the acceptance of boar meat differing in skatole and androstenone concentrations

The reactions of German (n=472) and Spanish (n=480) consumers of the European boar taint study to boar meat were evaluated according to their androstenone sensitivity. Consumers were checked with pure substance for their androstenone sensitivity. Eighteen per cent of the German and 31% of the Spanish participants were highly sensitive to androstenone, with a significantly higher proportion of women. For insensitive/mildly sensitive consumers, skatole level was the main determinant for the rating of flavour and odour of pork. For highly sensitive consumers, androstenone significantly influenced the rating of odour (Spain and Germany) and flavour (Germany) and is the predominant problem for that group, due to the high percentage of carcasses with high androstenone levels.     

Monitoring androstenone levels in boars by direct immunochemical analysis of serum samples

The possibility of using blood samples for screening high levels of boar taint steroid androstenone (5α-androst-16-en-3-one) was studied both in living animals at the farm and carcasses at the slaughterhouse. The steroid was measured from boar serum and fat samples with a simple time-resolved fluoroimmunoassay. Fat samples contained androstenone in the range of 90-7500 ng/g (n=214), and 74.8% of the samples exhibited fat androstenone levels above 500 ng/g. Androstenone concentrations in blood samples were measured by direct serum assay and ranged up to 215 ng/ml (n=214). The levels of androstenone were correlated (r=0.78-0.88, P<0.001) between the serum and fat samples obtained at slaughter and serum samples taken at the farm 7-11 days before slaughter. A direct serum analysis seems to give a reliable indication of the androstenone level in fat and it can also be used in the screening of living animals.