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.     

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.     

Reaction to boar odour by different South African consumer groups

A consumer sensory study of South African pork consumers (n=300) including equal numbers of males and females and consumers from three ethnic groupings (black, white and coloured consumers) was conducted to determine the effects of gender and ethnicity on consumer reactions towards boar odour compounds. Samples consisted of boar fat with varying concentrations of skatole and androstenone. Each consumer evaluated the odour of seven fat samples. Consumers’ liking of pork meat in general had a significant effect on the hedonic rating of the boar odour. The majority of consumers represented in this study would be dissatisfied with pork meat exhibiting detectable levels of skatole. In general, more females compared with males, will also respond more negatively towards samples with detectable levels of androstenone. An apparent liking for samples with medium levels of androstenone (0.5–1 μg/g) was found for some consumers, especially males, and can be partly attributed to the inability of some consumers to smell this compound at these levels or a genuine liking for the odour of androstenone. Significant differences in the sensitivity of consumers from different ethnic groups were found with white females responding more negatively than white males and blacks. Although it was not possible to compare responses directly with the white and black groups, it was found that a higher percentage of coloureds responded negatively to boar odour compounds. Coloured males responded particularly negatively towards samples with detectable skatole, while black males, in general, were found to be more critical than black females. Based on these results it is predicted that the majority of consumers would be less willing to consume pork meat exhibiting detectable levels of boar odour.     

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.     

Threshold Detection of Boar Taint Chemicals Using Parasitic Wasps

Abstract: Surgical castration has been long used to prevent consumers from experiencing taint in meat from male pigs, which is a large problem in the pig husbandry industry. Due to obvious animal welfare issues, the EU now wants an alternative for castration, suggesting an urgent need for novel methods of boar taint detection. As boar taint is only a problem when taint chemicals exceed a well‐defined threshold, detection methods should be concentration‐specific. The wasp, Microplitis croceipes’ ability to learn and respond to particular concentrations of the boar taint compounds, skatole, androstenone, and indole was tested. Also tested was the wasps’ ability to discriminate between known concentrations of indole, skatole, and androstenone in real boar fat samples at room temperature. Wasps were trained using associative learning by providing food‐deprived wasps with sucrose–water in the presence of specific odor concentrations. Trained wasps’ responses were tested to a range of concentrations of 3 compounds. Wasps showed unidirectional generalization of learned concentration responses, whereby the direction of concentration generalization was shown to be chemical‐dependent. Through both positive (sucrose) and negative feeding experiences (water only) with varying compound concentrations, the wasps can also be conditioned to respond to concentrations exceeding a defined threshold, and they were successful in reporting low, medium, and high concentrations of indole, skatole, and androstenone in boar fat at room temperature. The need for threshold detection rather than simple detection of absence/presence applies to many food quality issues, including the detection of spoilage or pest damage in crops or stored foods. Practical Application: An inexpensive and reliable means of detecting boar tainted pork at slaughter to avoid tainted meat on the market and dissatisfied consumers.     

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.     

Effect of sex grouping during growth on 5α-androstenone development in boars at three commercial slaughter weights

Groups of boars and gilts were grown in either single-sex or mixed groups to slaughter weights of 81, 93 or 110 kg liveweight to examine the effects of sex grouping on growth performance, animal behaviour and the development of androstenone-taint in the males. Results confirmed the well known growth advantages of entire males and showed that no husbandry problems of any significance were incurred with the boars and gilts raised together, particularly in those groups slaughtered before reaching 100 kg liveweight. In pigs taken to heavier weights, boars were successfully reared with gilts without incurring management problems, although growth performance was slightly poorer for the boar/gilt groups compared with mixed castrates and gilts, and some of the boars had high concentrations of androstenone in the fat. Thus, for pigs intended for slaughter at heavy commercial weight, the evidence suggests that split-sex rearing (and therefore feeding) should be practised. No useful correlations of androstenone concentration in fat were found with thickness of backfat, age, carcass weight or liveweight at slaughter.     

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.     

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±125ng/g and 1206±895ng/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 (175ng/g) compared to CT (92ng/g) and TT (93ng/g). Applying suggested sensory threshold levels for skatole (>150ng/g) and androstenone (>2000ng/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.     

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.     

An international study on the importance of androstenone and skatole for boar taint: I. Presentation of the programme and measurement of boar taint compounds with different analytical procedures

An international study, involving 11 participants in 7 European countries, was conducted to provide scientific evidence for an objective measurement of boar taint in entire male pigs and its possible variation between countries. The specific objectives were to determine the respective contributions of androstenone and skatole to boar taint and their possible variations according to production systems and consumer populations. Over 4000 entire male pigs and 200 gilts were raised and slaughtered in 6 countries. Meat samples were taken from the loin and backfat samples were used for the rapid measurement of androstenone and skatole. A sub-population of 377 entire males and 42 gilts was then selected in such a way as to represent all combinations of skatole and androstenone levels. Androstenone and skatole levels in the selected samples were checked, using established reference methods. Meat samples from the selected animals were used for sensory evaluation by trained panels and for consumer surveys in 7 European countries. The present paper gives a general presentation of the programme and reports the main characteristics of the samples. Three companion papers present the results of the evaluation by trained sensory panels [Dijksterhuis, G., Engel, B., Walstra, P., Font i Furnols, M., Agerhem, H., Fisher, K., Oliver, M. A., Claudi-Magnussen, C., Siret, F., Béague, M. P., Homer, D. B., & Bonneau, M. (2000). An international study on the importance of androstenone and skatole for boar taint: II. Sensory evaluation by trained panels in seven European countries. Meat Science54, 261-269], the results of the consumer surveys [Matthews, K. R., Homer, D. B., Punter, P., Béague, M. P., Gispert, M., Kemspter, A. J., Agerhem, H., Claudi-Magnussen, C., Fischer, K., Siret, F., Leask, H., Font i Furnols, M., & Bonneau, M. (2000). An international study on the importance of androstenone and skatole for boar taint: III. Consumer survey in seven European countries. Meat Science, 54, 271-283] and the main conclusions and recommendations [Bonneau, M., Walstra, P., Claudi-Magnussen, C., Kempster, A. J., Tornberg, E., Fischer, K., Diestre, A., Siret, F., Chevillon, P., Claus, R., Dijksterhuis, G., Punter, P., Matthews, K. R., Agerhem, H., Béague, M. P., Oliver, M. A., Gispert, M., Weiler, U., von Seth, G., Leask, H., Font i Furnols, M., Homer, D. B., & Cook, G. L. (2000). 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 out carcasses on the slaughter line, main conclusions and recommendations. Meat Science, 54, 285-295]. Seasonal effects and differences between countries in skatole and androstenone levels are presented elsewhere [Walstra, P., Claudi-Magnussen, C., Chevillon, P., von-Seth, G., Diestre, A., Matthews, K. R., Homer, D. B., & Bonneau, M. (in press). An international study on the importance of androstenone and skatole for boar taint: Levels of androstenone and skatole according to country and season. Livestock Production Science]. A supplementary paper considers the effects of human sensitivity to androstenone and skatole on the consumer acceptance of entire male pork [Weiler, U., Font i Furnols, M., Fischer, K., Kemmer, H., Oliver, M. A., Gispert, M., Dobrowolski, A., & Claus, R. (in press). 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. Meat Science]. A study of possible other compounds contributing to boar taint was also carried out within this programme.     

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.     

Effect of dietary chicory on boar taint

Following preliminary screening and feeding trials on farms supplying a commercial abattoir, 360 entire male pigs were used to evaluate the effects of different percentages of chicory (Cichorium intybus L.) on levels of boar taint compounds and sensory aspects in backfat. Pigs were fed 0, 3, 6 or 9% chicory in the diet, 30 pigs being sampled at 3 different times: initially to measure basal levels of skatole and androstenone and after 1 and 2 weeks on the test diets. Cooked samples of backfat were presented to a trained sensory panel for "sniff" tests. Chicory fed at 9% for 2 weeks reduced skatole levels significantly (P<0.001), with 0.55 of pigs below 0.05 μg/g, typical of levels in castrated males. Abnormal odour scores were significantly lower for pigs in this group compared with 0% pigs (P<0.001), however, androstenone concentration was significantly higher in this group after the 2 week feeding period (P<0.005). Thus, feeding 9% chicory for 2 weeks was effective in reducing backfat skatole concentrations and abnormal odour scores of cooked fat but not androstenone concentration.     

Influence of soiling on boar taint in boars

It has been suggested that skatole, one of the main compounds responsible for boar taint, can be lowered by keeping pigs clean, as skatole can be absorbed through skin and/or lungs (Hansen, Larsen, Jensen, HansenMoller & Bartongade, 1994). With this experiment, we further investigated this hypothesis by comparing extremely clean with extremely dirty animals with regard to the occurrence of boar taint. One group of boars was washed daily and pens were mucked on and littered down daily (CLEAN), a second group of boars was rubbed with faeces daily (DIRTY) and a third group of boars was kept in control conditions (CONTROL). The treatment was performed during the last four weeks before slaughter. According to the standardised consumer panel evaluations, boars subjected to extra soiling had a higher concentration of boar taint than boars that were kept extra clean. In contrast, expert panels judged general meat flavour to be inferior in CLEAN than CONTROL pigs. The home consumer panel, the hot iron method, and laboratory analyses, i.e., the presence of indole, skatole and androstenone in fat and serum, all showed no significant differences. So no clear indications towards skatole reduction by improving cleanliness of pigs were found.     

Comparison of heating methods and the use of different tissues for sensory assessment of abnormal odours (boar taint) in pig meat

Five heating methods (microwave, hotwire, boiling at 25 °C and 75 °C and melting) were used to generate cooking odours from backfat of entire male pigs and a 'composite' sample consisting of fat and muscle from the head along with salivary glands. The methods elicited significantly different scores for odours from 4 groups of 10 samples differing in their concentrations and ratios of skatole and androstenone. The odours (pork odour, abnormal odour, skatole odour and androstenone odour) were assessed by 3 experienced assessors. Correlations between skatole and androstenone concentrations and abnormal odour score in backfat were higher for skatole, suggesting it is the more important boar taint compound. In the composite sample, androstenone concentration was much higher than in backfat and androstenone was a more important contributor to boar taint. The microwave, hotwire and boiling (75 °C) methods produced the clearest separation between samples and the microwave method was considered the most suitable for on-line use.     

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.     

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.     

The measurement of the responses to different odour intensities of `boar taint' using a sensory panel and an electronic nose

This study explored the possibility of using an electronic nose (e-nose) with a 12-conducting-polymer sensor array combined with pattern recognition routines to discriminate between varying intensities of boar taint. A set of samples in a model system comprising a neutral lipid base containing varying combinations of androstenone and skatole were tested, as were pork fat samples. The e-nose responses for pork fat were calibrated against those given by a trained 10-member sensory panel for abnormal odour of the same samples from a total of 60 Large White cross-bred pigs. The e-nose responses related strongly to those of the sensory panel with a significant (p<0·01) canonical correlation of 0·78. The data set was used to develop a discriminant function for grouping pork samples into three `response classes': normal, doubtful and abnormal. Based on this, the e-nose identified all the abnormal samples correctly. However, 16% of the normal samples were also classified as abnormal. It was concluded that, in general, the electronic nose can discriminate between different levels of boar taint and that although a high specificity of sensors to androstenone and skatole may be desirable it may not be entirely important to the development and configuration of a boar taint sensor array.