Concentration of indoles and other rumen metabolites in sheep after a meal of fresh white clover,perennial ryegrass or Lotus corniculatus and the appearance of indoles in the blood

The objective of this study was to determine the effect of feeding three fresh forage diets, white clover (WC), perennial ryegrass (PRG) and Lotus corniculatus (LC), on the formation of indole and skatole in the rumen of sheep. The formation of indole and skatole in the rumen and their appearance in the blood were also compared. Peak rumen indole and skatole concentrations per kg crude protein intake (CPI) were significantly higher when feeding WC compared with PRG and LC (P < 0.05) and this was associated with a significantly higher rumen concentration per kg CPI of ammonia, branched chain volatile fatty acids, total nitrogen and soluble nitrogen (P < 0.05). Greater indole and skatole concentrations when feeding WC can be attributed to high solubility and rapid degradation of the forage protein. LC had a similar nutrient composition to WC, but the condensed tannins in LC slowed protein degradation and reduced indole and skatole formation. Indole and skatole concentrations peaked in the plasma 1–2 h after the end of feeding, indicating that skatole and indole are rapidly absorbed from the rumen into the blood. High indole and skatole formation with low intakes of WC indicates that the WC component of traditional New Zealand pastures may be the primary cause of undesirable pastoral flavours that result from the presence of indoles in meat. To ameliorate undesirable flavours, producers reliant on pastoral systems will need to consider using alternative forages such as LC to reduce protein solubility and degradation rate. Copyright © 2007 Society of Chemical Industry     

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     

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.     

Effect of different condensed tannin‐containing forages,forage maturity and nitrogen fertiliser application on the formation of indole and skatole in in vitro rumen fermentations

The objectives of this work were to establish the effect of seven different forages with a varied condensed tannin (CT) content, plant maturity and nitrogen fertiliser application on the ruminal formation of skatole and indole using an in vitro method designed to mimic rumen fermentation conditions. After 10 h of incubation, the concentration of indole and skatole was highest when incubating white clover (P < 0.05). Polyethylene glycol addition, to inhibit CT, showed that CT significantly reduced the formation of indole and skatole when incubating Lotus corniculatus, sulla, Dorynium rectum and Lotus pedunculatus (P < 0.01). Mature forage growth resulted in a significantly lower concentration of indole and skatole being formed in vitro compared to the incubation of new spring growth (P < 0.001). A higher application of nitrogen fertiliser to perennial ryegrass‐based pasture resulted in a higher crude protein concentration in the plant and a significantly higher concentration of skatole formed in vitro (P < 0.001). Forages containing CT reduced the conversion of endogenous protein to indole and skatole and plants containing a higher CT concentration tended to be more effective, but compositional differences of CT between forages may also have had an influence. New forage growth or swards that had a high application of nitrogen fertiliser promoted the formation of indole and skatole. Copyright © 2007 Society of Chemical Industry     

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.     

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.     

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.     

The effects of method of castration,rearing condition and diet on sensory quality of pork assessed by a trained panel

Beside surgical castration possible alternatives helping to reduce the incidence of boar taint in cooked pork are rearing conditions, immunocastration and feeding strategies for entire males known to lower the skatole levels. The goal of this study was to assess the effects of these alternatives on the sensory acceptability of pork. In experiment 1, carcasses from barrows, entire males (EM) and entire males fed raw potato starch (EM+) 7 d before slaughter were selected based on the androstenone (≤ 2 µg/g) and skatole (≤ 0.32 µg/g) levels. In experiment 2, carcasses from barrows, immunocastrates (IC), entire males either group-penned (EMG) or individually penned (EMP) were selected based on the aforementioned criteria. Boar odour and boar flavour intensities of longissimus dorsi (LD) and neck chops were evaluated by trained panellists. Boar odour and flavour scores were higher (P < 0.01) in neck than LD chops. Although skatole tissue levels in barrows and EM+ were similar (P > 0.05), boar odour and flavour scores were greater (P < 0.05) in EM+ than barrows. In experiment 2, scores for boar odour and flavour were lower (P < 0.05) in pork from barrows and IC than EMP, with intermediate values for EMG. In conclusion, we observed a discrepancy between the known boar taint compounds androstenone and skatole and sensory acceptability, which indicates that other factors influenced the perception of boar taint. Thus, surgical castration with or without anesthesia or immunocastration are still the safest methods to avoid boar taint.     

Nutritional and sensory aspects of light lamb meat enriched in n−3 fatty acids during refrigerated storage

Fatty acid content and sensory characteristics of meat from light lambs fed three diets supplemented with different sources of n−3 fatty acids (fish oil, extruded linseed and extruded linseed plus microalgae) and a control diet during refrigerated storage have been studied. The meat from lambs fed linseed diets had the highest levels of C18:3 n−3, while animals fed fish oil had the highest long-chain n−3 polyunsaturated fatty acids (PUFA). Thus, 100 g of meat from lamb fed the fish oil diet provided 183 mg of long-chain n−3 PUFA, representing 40% of the daily recommended intake. The levels of n−3, n−6 and long-chain n−3 PUFA decreased during a 7-day storage period. The meat from lambs fed the fish oil diet had high scores of fish odour and flavour and rancid odour and flavour and the lowest overall liking. Rancid odour and flavour increased after storage, mainly in supplemented groups.     

Effect of branched-chain fatty acids, 3-methylindole and 4-methylphenol on consumer sensory scores of grilled lamb meat

Tenderness, flavour, overall liking and odour are important components of sheepmeat eating quality. Consumer assessment of these attributes has been made for carcasses from the Information Nucleus Flock (INF) of the Cooperative Research Centre for Sheep Industry Innovation. The concentrations of three branched chain fatty acids, 4-methyloctanoic (MOA), 4-ethyloctanoic (EOA) and 4-methylnonanoic acids (compounds related to ‘mutton flavour’ in cooked sheepmeat) and 3-methylindole and 4-methylphenol (compounds related to ‘pastoral’ flavour) were determined for 178 fat samples taken from INF carcasses. Statistical modelling revealed that both MOA and EOA impacted on the ‘Like Smell’ consumer sensory score of the cooked meat product (P < 0.05), with increasing concentration causing lower consumer acceptance of the product. None of the compounds though had an effect on the liking of flavour. Obviously, reducing the effect of MOA and EOA on the odour of grilled lamb will improve consumer acceptance of the cooked product but other factors affecting the eating quality also need to be considered.     

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 pasture species on the flavour, odour and keeping quality of lamb and mutton

Lambs fed white clover species (Trifolium repens cv. Grasslands Huia or cv. Grasslands 4700) showed a stronger flavour in the fat and lean and a greater intensity of odour in the casseroled 12th rib chop than lambs fed on perennial ryegrass (Lolium perenne). The results for hoggets generally supported those for lambs. Differences in flavour of the lean of the lambs appeared within three weeks from the beginning of the experiment. Differences in intensity of flavour and odour between the shoulder, loin and leg of the hoggets were small. After storage for 8 months at —15°, the thiobarbituric acid (TBA) values of the lean and fat of the 12th rib chop were highly significantly greater in lambs fed white clover than in those fed perennial ryegrass. Similar differences were found in hoggets using carcasses that had been stored for one to two months. In the hoggets, significantly higher TBA values were found in the fatty tissues of the leg than in the loin and shoulder. Differences between the TBA values of the lean were not significant.     

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.     

Diet and growth effects in panel assessment of sheepmeat odour and flavour

The effects of sheep age and diet on several odours and flavours are described. Ram lambs raised on ewe's milk then a corn-based diet were compared with lambs raised on milk and a pasture of grass/clover, six treatments in all. A seventh treatment comprised very old ewes maintained on pasture. Fat and lean from forequarters was minced and cooked together. Cooked lean was assessed for intensity by a sensory panel for 10 flavour attributes. Four showed significant (P < 0.01) treatment effects: 'sheepmeat', 'animal', 'liver', and 'poultry'. Sheepmeat flavour was highest in the slow-grown pasture-fed lambs. Animal flavour-the flavour associated with the odour of confined livestock-showed a similar pattern with treatment. Liver flavour was highest in ewe meat, and the biochemical origin of this flavour is discussed. Eleven related odour attributes were assessed on the rendered fat with a novel olfactometer. Five attributes showed highly significant treatment effects for intensity (P < 0.001): animal and sheepmeat odours showed a similar distribution to the equivalent flavours; likewise cabbage and rancid odours were associated with the two slow-grown pasture treatments. A comparison of the odour and flavour statistics showed that the sense of smell was the more discriminating in sheepmeat assessment, and also confirmed that fat was the true source of sheepmeat odour/flavour. In respect of sheepmeat production for effective marketing, the data showed that at 90 days, a pastoral diet resulted in slightly enhanced odours when compared with a corn-based diet. By 215 days, however, many undesirable odours were exacerbated. Since these older rams were more sexually developed, a sex rather than an age effect could not be excluded. Rendered fat from this work was further used in a companion study (Yang et al., 1997. Meat Sci., 45, 183-200) in an attempt to link individual volatile compounds to odour attributes.     

Effect of concentrate feeding on instrumental meat quality and sensory characteristics of fallow deer venison

Venison from twenty four hybrid fallow deer does, 36 months old with an initial body condition score (BCS) of 2, was tested to determine the influence of feed type on meat quality. Feeding with concentrates increased BCS (P < 0.01) but did not affect ultimate pH (P > 0.05). BCS 4 animals had higher intra muscular fat (IMF) (P < 0.01), and more tender meat (P < 0.05). Venison from does fed over 24 weeks exhibited less redness (P < 0.01) than those fed for 19 weeks regardless of feed type.Panellists evaluated samples for colour, flavour, tenderness, juiciness and overall liking. They detected significantly (P < 0.05) stronger flavour in meat from animals fed concentrates. Male panellists detected flavour differences within meat from animals fed concentrates (P < 0.05), with longer feeding periods resulting in stronger flavour. There was no difference in overall liking, therefore finishing fallow deer on grain-based concentrate feeds prior to slaughter provided little commercial advantage.     

Influence of aromatic components from pig manure on odour and flavour of cooked chicken meat

An experiment with 96 broilers distributed across six treatments was carried out. Each treatment consisted of four cages each with four broilers. The six treatments were: the control (treatment 1), broilers placed in cages with a welded wire bottom hanging over smelling pig manure (treatment 2), or placed on a pig manure mat (treatment 3), combined with two intervals, i.e. 1 week (group a) and 2 weeks (group b) before slaughter. The content of skatole in the abdominal fat was measured, and the odour and flavour impressions after cooking were evaluated by a taste panel. The treatments were compared with two control treatments: broilers produced traditionally in cages either without (treatment 1a) or with (treatment 1b) contact with their own manure for a fortnight. A higher skatole level in abdominal fat of broilers in physical contact with pig manure (treatment 3) was measured, compared with broilers without physical contact with pig manure for at least 1 week (treatments 1 and 2) (p<0.05). The consumer taste panel evaluation was not affected by the fact that the broilers had been lying on pig manure. However, the evaluation of the odour of the cooked meat, when opening the cooking bag, was to some extent negatively affected by the experimental treatments of broilers, which had physical contact with the pig manure (p<0.05). Furthermore, physical contact with the manure, regardless of whether it was broiler or pig manure, proved to affect the odour of the meat (p<0.01). For broilers placed in cages without physical contact with the pig manure (treatments 2a and 2), the smell had no negative influence on either the odour or on the flavour of the meat, and the content of skatole was not different from that of the broilers on the control treatment. A taste panel evaluation performed on four broilers of the treatments 1a, 2b, and 3b showed no flavour differences among treatments. Whether the broilers had been exposed to a given treatment for 1 or 2 weeks made no difference, either to the skatole concentration or to the odour or flavour impression of the newly cooked chicken meat.     

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.     

Screening of indoles in cheese

 Indole and skatole (3-methylindole) are formed from tryptophan by microbial activity. Depending on their concentration, they may either contribute to an unpleasant odour, as described for pig meat, or alternatively to a positive aroma profile, as during ripening of cheese. In a screening study, the two indoles in various types of cheese were determined, by both HPLC and GC. It was found that the two compounds mainly occur in mould cheese, such as Camembert and are even more pronounced in blue-veined cheese. The concentrations in unripened cheese were up to 700 ng/g fat for indole and 50 ng/g fat for skatole. The formation of the two indoles increases when proteolysis during ripening provides more tryptophan. Skatole formation appears to be specifically favoured by a low pH and anaerobic conditions. Received: 14 May 1997