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.     

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.     

Marinating as a technology to shift sensory thresholds in ready-to-eat entire male pork meat

This study investigated the effect of marinades in improving the eating quality in ready-to-eat boar meat. Neck chops with fat content below 18.9%, skatole 1.1 ppm (range 0.03–1.1) and androstenone 5.6 ppm (range 0.01–5.6) were used. In a screening experiment different marinades were tested for their ability to mask boar taint (defined as manure and urine odour and flavour). Liquid smoke and oregano extracts appeared to have the best potential for masking, and were studied in detail. Results from the study indicated that marinated chops with skatole content of approximately 0.4 ppm appeared similar to castrates in boar taint. Chops with skatole contents above 0.7 ppm remained unmasked despite the use of strongly flavoured marinades. Unmarinated chops served at 60 °C were more tainted than those served at 15 °C, but scored lower for boar taint when reheated, although the concentrations of androstenone and skatole remained the same. The fat content of the chops was not well correlated to the perception of boar taint. The attributes manure and urine were correlated with the level of skatole, but urine attribute was not a good indicator of the androstenone level.     

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.     

Marinating as a technology to shift sensory thresholds in ready-to-eat entire male pork meat

This study investigated the effect of marinades in improving the eating quality in ready-to-eat boar meat. Neck chops with fat content below 18.9%, skatole ⩽1.1 ppm (range 0.03–1.1) and androstenone ⩽5.6 ppm (range 0.01–5.6) were used. In a screening experiment different marinades were tested for their ability to mask boar taint (defined as manure and urine odour and flavour). Liquid smoke and oregano extracts appeared to have the best potential for masking, and were studied in detail. Results from the study indicated that marinated chops with skatole content of approximately 0.4 ppm appeared similar to castrates in boar taint. Chops with skatole contents above 0.7 ppm remained unmasked despite the use of strongly flavoured marinades. Unmarinated chops served at 60 °C were more tainted than those served at 15 °C, but scored lower for boar taint when reheated, although the concentrations of androstenone and skatole remained the same. The fat content of the chops was not well correlated to the perception of boar taint. The attributes manure and urine were correlated with the level of skatole, but urine attribute was not a good indicator of the androstenone level.     

Effect of feeding fermentable fibre-rich feedstuffs on meat quality with emphasis on chemical and sensory boar taint in entire male and female pigs

Skatole, androstenone and other compounds such as indole cause boar taint in entire male pork. However, female pigs also produce skatole and indole. The purpose of this experiment was to minimise boar taint and increase overall impression of sensory quality by feeding entire male and female pigs with fibre-rich feedstuffs. The pigs have been fed three organic diets for either 1 or 2 weeks prior to slaughter of which two diets contained different fermentable fibre-rich feedstuffs – 10–13.3% dried chicory roots or 25% blue lupines. These two treatments were compared with pigs fed with an organic control diet for either 1 or 2 weeks prior to slaughter. Lupines significantly reduced skatole in blood and backfat for both genders after 1 week. Moreover, lupines showed negative impact on growth rate and feed conversion whilst chicory showed no significant differences in this respect. However, the indole concentration was significantly lower in chicory than lupine fed pigs. From a sensory perspective, chicory and lupine feeding reduced boar taint since odour and flavour of manure related to skatole and urine associated to androstenone were minimised. The level of boar taint in the entire male pigs was most effectively reduced after 14 days by both fibre-rich feeds while lupine had the largest influence on “boar” taint reduction in female pigs.     

Sensory characterization of meat from pigs vaccinated against gonadotropin releasing factor compared to meat from surgically castrated,entire male and female pigs

Boar taint is a sensory defect mainly due to androstenone and skatole. The most common method to control boar taint is surgical castration at an early age. Vaccination against gonadotropin releasing factor (also known as immunocastration) is an alternative to surgical castration to reduce androstenone content. In this experiment, loins from 24 female (FE), 24 entire male (EM), 24 vaccinated males (IM) and 23 surgically castrated males (CM) were evaluated by eight trained panellists in 24 sessions. Loins were cooked in an oven at 180 °C for 10 min. Furthermore loins were evaluated by consumers and its androstenone and skatole content were also chemically determined. Meat from EM had higher androstenone and skatole odour and flavour than meat from FE, IM and CM and lower sweetness odour scores. High correlations were found between androstenone and skatole levels assessed by trained panelists, chemical analysis and consumers’ acceptability. Moreover meat from EM is mainly related to androstenone and skatole attributes.     

Effect of feeding fermentable fibre-rich feedstuffs on meat quality with emphasis on chemical and sensory boar taint in entire male and female pigs

Skatole, androstenone and other compounds such as indole cause boar taint in entire male pork. However, female pigs also produce skatole and indole. The purpose of this experiment was to minimise boar taint and increase overall impression of sensory quality by feeding entire male and female pigs with fibre-rich feedstuffs. The pigs have been fed three organic diets for either 1 or 2 weeks prior to slaughter of which two diets contained different fermentable fibre-rich feedstuffs – 10–13.3% dried chicory roots or 25% blue lupines. These two treatments were compared with pigs fed with an organic control diet for either 1 or 2 weeks prior to slaughter. Lupines significantly reduced skatole in blood and backfat for both genders after 1 week. Moreover, lupines showed negative impact on growth rate and feed conversion whilst chicory showed no significant differences in this respect. However, the indole concentration was significantly lower in chicory than lupine fed pigs. From a sensory perspective, chicory and lupine feeding reduced boar taint since odour and flavour of manure related to skatole and urine associated to androstenone were minimised. The level of boar taint in the entire male pigs was most effectively reduced after 14 days by both fibre-rich feeds while lupine had the largest influence on “boar” taint reduction in female pigs.     

Influence of volatile compounds on the development of off-flavours in pig back fat samples classified with boar taint by a test panel

Dynamic Head Space methodology was applied to evaluate the possible contribution of some volatile compounds to the development of boar taint in pig backfat samples with low concentrations of skatole and androstenone, but which had previously been classified as tainted by a trained test panel. Volatile compounds were collected in a trap of graphited charcoal and analysed by GC–MS in Scan mode. Aldehydes and short chain fatty acids, compounds that play a significant role in the development of undesirable aromas in food products, were the main classes of compounds identified in this study, although the possible contribution of other compounds that were detected in a minor proportion – such as alcohols and ketones – was evaluated. Styrene and 1,4-dichlorobenzene, compounds that may have come from an external contamination, showed a high concentration in the samples classified with boar taint, so these compounds could have been responsible for the development of some off-flavours in the fat samples studied in this work. In the same study, skatole and androstenone were also determined by normal phase HPLC and GCMS, respectively.     

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.     

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.     

A human nose scoring system for boar taint and its relationship with androstenone and skatole

A system for sensory evaluation of boar taint was used to evaluate boar taint in fat samples from 6574 entire males. The term "human nose scoring" has been used to describe this system. The samples from each boar were heated with a hot iron and three panelists assigned scores of 0 to 4. The reproducibility of HNS ranged from 0.19 to 0.32 reflecting natural variation in the ability of human beings to detect different odors. The correlations of HNS with androstenone ranged from 0.22 to 0.52, while those with skatole ranged from 0.31 to 0.89, suggesting that skatole is a better predictor of boar taint. Considering (1) the relationship of HNS with the boar taint compounds, (2) the ability of HNS to capture variation not accounted for by the boar taint compounds, (3) low estimation costs and (4) low time requirements, HNS can be used in large scale evaluations of boar taint.     

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.     

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.     

The effects of castration on the eating quality of dry-cured ham

The influence of the castration of entire male pigs on the eating quality of dry-cured ham was evaluated. Forty-eight dry-cured hams (435-day aged) were studied from entire and castrated males of two different crossbreeds. The proximate composition of the meat and the androstenone, indole and skatole fat content were determined. The entire hams were classified according to the androstenone and skatole content. Sensory analysis was carried out by a trained panel, evaluating marbling, juiciness, saltiness, graininess, toughness, overall flavour, boar odour and boar flavour. Also carried out was a preference and acceptability paired test by consumers. Castration increased meat fattening and reduced the androstenone and skatole levels of the fat. Significant differences (P<0.05) were found between entires and castrates for the average values given in all the sensory attributes studied. The dry-cured ham from castrates was scored as more flavoured, more marbled and softer. It was also perceived as less grainy, less salty and having less boar odour and flavour. The sensory perception of boar odour was more intense than that of flavour in dry-cured ham and appears to be related to the level of androstenone and skatole in fat. Dry-cured ham from castrated males was also more accepted and more preferred by consumers, especially women and habitual consumers. Castration of male pigs contributes to improve the quality of dry-cured ham. The rejection caused by boar odour and flavour is reduced, improving the overall flavour, texture and juiciness. In addition, the saltiness is less pronounced in ham from castrates. For this reason, the production of high quality dry-cured ham will have to shoulder the extra costs associated with processing castrated pigs.     

Quantification of myosin heavy chain isoform in porcine muscle using an enzyme-linked immunosorbent assay

The aim of the present study (part of an EU AIR programme on boar taint) was to make objective the perception of boar taint in entire male pork, and to relate the perception to skatole and androstenone levels. Trained analytical sensory panels in seven European countries assessed pig meat with known levels of androstenone and skatole. The panels performed a sensory profiling using the attributes pig, urine, manure/stable, naphthalene/mothballs, rancid, sweet, sweat and abnormal, both for odour and flavour in separate sessions. It turned out to be difficult to harmonise sensory methodology for seven sensory panels throughout the EU, especially with respect to the exact level of training the panellists received. Sensory panels in general were able to differentiate between the two compounds and between different levels of the compounds, though substantial differences between the panels in the different countries existed. Androstenone was found to relate mostly to the urine attribute, while skatole related mostly to manure and, to a lesser extent, to naphthalene.     

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.     

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.     

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.