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.     

Method for the determination of indole and skatole in pig fat

A quantitative method was developed to determine by gas-liquid chromatography the concentrations of indole and skatole in pig fat. Methanolic extracts of subcutaneous adipose tissue were de-fatted by cooling, concentrated using solid-phase extraction columns, and analysed in a gas chromatograph fitted with a polar capillary column and a flame ionisation detector. The method detects indole and skatole to below 0-002 mg kg^?1 fat.     

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.     

Evaluation of the contribution of skatole, indole, androstenone and androstenols to boar-taint in back fat of pigs by HPLC and capillary gas chromatography (CGC)

Fifteen samples of fat with boar taint were selected by means of a trained panel-test, from 200 carcasses of entire male pigs, and they were analyzed to determine skatole, indole, 5-Androst-16-en-3-one (5-An), 5-Androst-16-en-3-ol (5An3) and 5-Androst-16-en-3β-ol (5An3β).

The extraction was carried out with methanol, the extract was placed at −20°C during 10 min to eliminate the fat and after evaporation of methanol a Florisil clean up was applied to obtain skatole and indole in a separate fraction from 5An and androstenols.

Skatole and indole were determined by HPLC in normal phase with a Spherisorb-NH₂ column and 5An and androstenols were analyzed by capillary gas chromatography (CGC). Selective procedures were used for detection: ratio at 220 nm and 280 nm (skatole/indole) and CGC coupled to a mass spectrometer working in the SIM (selected ion monitoring) mode at m/z 272 (o5An) and m/z 274 (androstenols). The results obtained showed that skatole and 5An concentrations exceeded that typical thresholds values in more than 50% of samples analyzed, suggesting a similar influence on boar taint. 5An3β showed higher concentrations than 5An3, but lower than 5An, exceeding only 1·00 μg/g in three samples.     

An international study on the importance of androstenone and skatole for boar taint: III. Consumer survey in seven European countries

There has long been debate over the relative importance for consumer acceptability of androstenone and skatole. The objective of this study was to investigate consumer reaction to pig meat of varying levels of androstenone and skatole in seven member states of the European Union. Over 4000 entire male pigs (and 233 gilts as a control) were produced in six European countries, and a sub-sample of 420 were selected to represent a range of androstenone and skatole levels. Loin samples from these pigs were distributed to the seven participating countries. Samples were prepared and cooked according to standardised procedures in each of the countries and presented to pork consumers. Each consumer rated samples from five pigs for liking of flavour and liking of odour. Although there were significant differences between country panels in their liking of meat from entire males generally, there was a greater degree of dislike as both androstenone and skatole increased. This reaction was stronger for skatole than androstenone and consumers reacted more negatively in terms of odour than flavour.     

Effects of raw potato starch and live weight on fat and plasma skatole,indole and androstenone levels measured by different methods in entire male pigs

A total of 96 entire male pigs were used to evaluate the effects of feeding raw potato starch (RPS) and live weight (LW; 90, 100 or 115 kg) on skatole, indole and androstenone levels in fat and plasma. Different methods to analyse the compounds were compared. Fat levels of skatole analysed by HPLC and colorimetric methods, and androstenone analysed by HPLC and ELISA, were highly correlated. However, androstenone values obtained by ELISA were overestimated. Feeding RPS reduced skatole levels in plasma and fat but did not affect indole levels. Androstenone levels in plasma measured by direct ELISA were not altered by diet, whereas the levels measured by ELISA with extraction were lower in the pigs fed RPS. Feeding RPS did not reduce androstenone levels in fat. Androstenone levels in fat were higher at 115 than at 90 kg, whereas skatole and indole levels were not affected by LW.     

Effects of dietary components including garlic on concentrations of skatole and indole in subcutaneous fat of female pigs

The results reported here showed that threshold concentrations of skatole and indole in rice-bran oil for Singaporean consumers were 0.028 μg/g and 0.051 μg/g, respectively, and that skatole and indole levels in subcutaneous fat of pigs can be affected by diet. In Experiment A, 31 female pigs were fed with diets based on plant products only (P) or plant plus animal by-products (AP), with added levels of garlic essential oil from zero to 2.15 g/kg feed. Concentrations of skatole and indole increased with increasing garlic concentration (P < 0.001). In Experiment B, P and AP diets were fed to 47 female pigs with different dietary lipid sources (fish oil, tallow, and a mix of linseed oil and soya oil). Skatole and indole concentrations were higher in backfat of pigs fed with the AP diet (P < 0.05), but were unaffected by the type of lipid.     

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.     

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.     

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.     

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.     

Physiological aspects of androstenone and skatole formation in the boar-A review with experimental data

The advantages of boars in fattening performance and carcass traits when compared with the other sexes are explained by testicular anabolic hormones. The steroid androstenone with a pronounced urine-like odour is the main objection against boar meat. More recently skatole (faecal odour) has been identified as another contributor to off-odour of pork. Androstenone is synthesized in the testes, secreted into the circulation and accumulated in adipose tissue due to its lipophilic property. Its biosynthesis is linked to the synthesis of anabolic testicular hormones. Therefore no practical method is available to maintain the sex-dependent anabolic potential of boars and to suppress androstenone selectively. Skatole is formed from tryptophan by specialized microbes in the colon when energy in the colon is limited. Gonadal hormones, but also growth hormone and IGF-1, favour its formation. Oestrogens, which are synthesized in high amounts in boar testes, decrease voluntary food intake, thus lowering the intestinal passage rate. Additionally oestrogens probably influence directly intestinal contractions via specific gut receptors. It appears, however, that glucocorticoids are more important. They counteract mitogenic hormones, such as IGF-1, ultimately leading to gut mucosal cell degradation. The resulting cell debris probably is the main source of tryptophan for microbial skatole formation. In contrast to androstenone, skatole formation can be easily suppressed by dietary means.     

Skatole and indole concentrations in Longissimus dorsi and fat samples of pigs

A HPLC-NP (normal phase-high performance liquid chromatography) method for determining the concentration of skatole and indole in Longissimus dorsi samples is described. Lipids containing skatole and indole were extracted in chloroform:methanol (2:1) at room temperature and dehydrated by liquid-liquid extraction with an aqueous solution saturated with 10% of sodium chloride. The organic phase was evaporated to dryness and redissolved in 10 ml of hexane:2-propanol (92:8). Indolic compounds were separated on a Hypersil aminopropylsilica column (5 μm) (250×4.6 mm i.d.). The mobile phase was hexane:2-propanol (92:8) and detection was by fluorescence (excitation at 280 nm and emission at 360 nm). Linearity was found in the range of 0.05-0.4 μg/g and the coefficient of correlation was above 0.99 for both compounds. The within day (n=5) variation was at 0.05, 0.2 and 0.4 μg/g and the CV (coefficient of variation) values for relative areas determined at these concentrations were less than 13%. This method was used to compare the concentrations of skatole and indole in different samples: L. dorsi muscle, the fat covering the L. dorsi and subcutaneous fat. A correlation was observed between the concentration of indole and skatole in the back fat and fat covering the L. dorsi samples (P<0.001, r=0.99). No significant correlation was obtained in L. dorsi samples, between skatole and indole levels. In spite of the correlation shown between skatole and indole concentrations in the back fat and L. dorsi samples, the mean concentrations of these compounds were significantly higher (P<0.05) in the back fat samples.     

Influence of fresh alfalfa supplementation on fat skatole and indole concentration and chop odour and flavour in lambs grazing a cocksfoot pasture

We investigated the influence of the level of fresh alfalfa supplementation on fat skatole and indole concentration and chop sensory attributes in grazing lambs. Four groups of nine male Romane lambs grazing a cocksfoot pasture were supplemented with various levels of alfalfa for at least 60 days before slaughter. Perirenal fat skatole concentration was higher for lambs that consumed alfalfa than for those that consumed only cocksfoot. The intensity of ‘animal’ odour in the lean part of the chop and of ‘animal’ flavour in both the lean and fat parts of the chop, evaluated by a trained sensory panel, increased from the lowest level of alfalfa supplementation onwards and did not increase further with increasing levels of alfalfa supplementation. The outcome of this study therefore suggests that these sensory attributes may reach a plateau when perirenal fat skatole concentration is in the range 0.16–0.24 μg/g of liquid fat.     

Controlling the formation of indole and skatole in in vitro rumen fermentations using condensed tannin

Indole and skatole are formed in the rumen from the fermentation of tryptophan and have been correlated to the presence of undesirable pastoral flavours in meat from grazing ruminants. A series of four in vitro rumen fermentation experiments was carried out to determine the effectiveness of condensed tannin (CT) for reducing the formation of indole and skatole. Experiment 1 utilised fresh white clover (WC; Trifolium repens) in ratios with the CT‐containing forage Lotus pedunculatus (LP; 97 g CT kg^?1 dry matter (DM)). Increasing the ratio of LP to WC decreased the formation of indole and skatole. Experiments 2 and 3 used extracts of CT from LP and grape seed added to incubations of fresh non‐CT‐containing WC or perennial ryegrass (PRG; Lolium perenne). Including the CT extracts in incubations at 40 and 80 g kg^?1 DM was more effective at reducing indole and skatole formation than lower concentrations of CT extract (P < 0.05). Including fresh LP in ratios with WC gave a linear decrease in indole and skatole concentration (indicating dilution), while including a CT extract in the incubations gave an exponential decrease in indole and skatole concentration (suggesting binding). Experiment 4 elucidated the mechanisms behind the action of CT by delayed addition of tryptophan and polyethylene glycol (PEG) to incubations of LP. Rumen microbes that were exposed to CT of LP in planta for up to 6 h and then provided with tryptophan were still able to convert tryptophan to indole and skatole. Adding PEG to incubations of LP after 6 h inhibited the activity of plant CT and increased the availability of substrate for indole and skatole formation. These studies have shown that a higher concentration of CT is more efficient for reducing indole and skatole formation and that CT contained within plants acts differently in in vitro rumen fermentations than additions of extracted CT. Under the conditions of these experiments, there was no evidence that CT contained in LP affected the protein present in WC in a mixed fermentation. Copyright © 2007 Society of Chemical Industry     

The diffusion of salt in pork muscle and fat tissue

The diffusion coefficients of salt in pork longissirnus dorsi muscle between –2 and 25° and subcutaneous back fat at ?2° have been determined and compared with coefficients calculated from reported results of other workers. The rate of salt diffusion did not depend on the muscle fibre direction. Freezing the muscle at ?20° had no effect on the diffusion coefficient subsequently determined at ?2°.     

肌内脂肪与背最长肌肉质、脂肪酸组成的关系

研究肌内脂肪（IMF）含量与肉质以及脂肪酸含量和组成比例之间的关系。以北京黑猪背最长肌（n=20）为实验材料,测定肌内脂肪含量、肉质以及脂肪酸含量和组成。结果显示,肌内脂肪含量与滴水损失呈显著正相关（p<0.05）,与蒸煮损失、剪切力、肉色的相关性不显著（p>0.05）。随着肌内脂肪含量升高,除长链多不饱和脂肪酸(C_20:3n6、C_20:4n6)以外的大多数脂肪酸含量增加,相对比例也上升;多不饱和脂肪酸的含量增加,但是相对比例下降;长链多不饱和脂肪酸的含量和相对比例都降低。随着肌内脂肪含量的升高,饱和脂肪酸（SFA）和单不饱和脂肪酸（MUFA）的含量升高,回归方程斜率分别为197.3和248.31,多不饱和脂肪酸（PUFA）增加的幅度不大,回归方程的斜率为11.43。结果提示:肌内脂肪含量较低时（本实验平均值2.86%）,对肉质的影响不显著。随着肌内脂肪含量升高,总脂肪酸含量（TFA）增加,主要归因于SFA和MUFA含量的增加。PUFA的含量相对稳定,受肌内脂肪含量变化的影响较小。