Molecular cloning, expression and functional characterization of the cytochrome P450 2A6 gene in pig liver

Raising intact male pigs would have a significant economic impact on the pork industry because intact males have improved feed efficiency and a greater lean yield of the carcass compared with barrows. However, the presence of skatole, a major cause of boar taint, in meat from intact male pigs could be highly objectionable to consumers. It has been shown that CYP2A6 is a key enzyme in the hepatic metabolism of skatole and that the activity of CYP2A6 is negatively correlated with skatole accumulation in fat. The aim of this study was to isolate and characterize CYP2A6 from pig liver, as well as identify genetic polymorphisms in the CYP2A6 gene, and examine the association between these polymorphisms and skatole level. We identified a single base deletion in CYP2A6, resulting in a frame shift in the coding region that produces a non-functional enzyme, which was associated with high levels of skatole in fat tissue.     

Cloning and mapping of the porcine cytochrome-p450 2E1 gene and its association with skatole levels in the domestic pig

The porcine cytochrome-p450 2E1 (CYP2E1) gene was isolated by screening a pig BAC library and partially sequenced. This sequence information was used to identify six single nucleotide polymorphisms (SNPs) within the CYP2E1 gene and its promoter. In addition, a microsatellite marker tightly linked to the CYP2E1 gene was subcloned from the BAC. One of these markers was used to map the CYP2E1 gene distal of SWC27 on SSC14, well outside reported quantitative trait loci on SSC14 for skatole, indole and taste test measures of boar taint. However, in a population of commercial pigs scored for backfat skatole levels, there was evidence of association between a SNP in the CYP2E1 promoter and skatole deposition, although there was no significant association between this SNP and skatole levels in the experimental cross.     

Biochemical, nutritional and genetic effects on boar taint in entire male pigs

Pork odour is to a great extent affected by the presence of malodorous compounds, mainly androstenone and skatole. The present review outlines the current state of knowledge about factors involved in the regulation of androstenone and skatole in entire male pigs. Androstenone is a pheromonal steroid synthesised in the testes and metabolised in the liver. Part of androstenone accumulates in adipose tissue causing a urine-like odour. Skatole is produced in the large intestine by bacterial degradation of tryptophan and metabolised by hepatic cytochrome P450 enzymes and sulphotransferase. The un-metabolised part accumulates in adipose tissue, causing faecal-like odour. Androstenone levels are mostly determined by genetic factors and stage of puberty, whereas skatole levels in addition to genetic background and hormonal status of the pigs are also controlled by nutritional and environmental factors. To reduce the risk of tainted carcasses entering the market, male pigs are surgically castrated in many countries. However, entire males compared to castrates have superior production characteristics: higher growth rate, better feed efficiency and leaner carcasses. Additionally, animal welfare aspects are currently of particular importance in light of increasing consumers' concerns. Nutrition, hormonal status, genetic influence on boar taint compounds and the methods to develop genetic markers are discussed. Boar taint due to high levels of skatole and androstenone is moderately heritable and not all market weight entire males have boar taint; it should thus be possible to select for pigs that do not have boar taint. In these studies, it is critical to assess the steroidogenic potential of the pigs in order to separate late-maturing pigs from those with a low genetic potential for boar taint. A number of candidate genes for boar taint have been identified and work is continuing to develop genetic markers for low boar taint. More research is needed to clarify the factors involved in the development of boar taint and to develop additional methods to prevent the accumulation of high concentrations of skatole and androstenone in fat. This review proposes those areas requiring further research.     

Skatole metabolites in urine as a biological marker of pigs with enhanced hepatic metabolism

Boar taint is a quality defect in meat, related to accumulation of skatole and androstenone in male pigs. The levels of skatole and its main metabolites in plasma and urine samples were measured with a validated liquid chromatography-MS method and related to activity of hepatic cytochrome P450 (CYP450) in order to identify ‘fast metabolizing’ pigs. Urine (n=46), blood (n=12), liver (n=25) and adipose tissue (n=46) were sampled from a total of 46 entire male pigs. Skatole levels in fat were negatively correlated to CYP2E1 activity and positively to 3-hydroxy-3-methyloxindole (HMOI), indole-3-carboxylic acid (ICA) and 2-aminoacetophenone in urine. HMOI and ICA levels in urine were the best predictors of high skatole levels in fat. In summary, the present study provided further evidence for the key role of CYP2E1 in skatole metabolism and suggested that measurement of HMOI and/or ICA in urine might provide information about skatole levels in live pigs.     

Characterisation of androstenone metabolism in pig liver microsomes

Androstenone (5 alpha-androst-16-en-3-one) is a steroid pheromone produced in the testis. Excessive accumulation of androstenone together with skatole (3-methyl-indole) in the adipose tissue of some male pigs leads to "boar taint". In isolated pig hepatocytes androstenone represses the expression of cytochrome P450IIE1 (CYP2E1), the enzyme principally responsible for skatole metabolism. Androstenone can be metabolised in liver microsomes but the pathway has not been established. We have investigated androstenone metabolism in liver microsomes from two breeds of pigs exhibiting low and high levels of androstenone in adipose tissue-Large White (LW) and Meishan (M), respectively. Androstenone was reduced in isolated liver microsomes mainly to beta-androstenol using NADH as a co-factor. The rate of beta-androstenol formation in the presence of NADPH was very low. In microsomes from LW pigs the rate of beta-androstenol formation from androstenone was six times higher than in M pigs. 3beta-hydroxysteroid dehydrogenase (3beta-HSD) was investigated as a likely candidate for the enzyme catalysing androstenone reduction in pig liver. RT-PCR analysis showed that there was no sequence difference in the cDNA encoding 3beta-hydroxysteroid dehydrogenase from LW and M pigs. However, competitive RT-PCR analysis showed that the expression of 3beta-hydroxysteroid dehydrogenase mRNA was about 12 times higher in the case of LW compared to M pigs. It is concluded that the rate of androstenone metabolism in pig liver microsomes is determined by the level of expression of hepatic 3beta-hydroxysteroid dehydrogenase. The differential expression of this enzyme could be a factor affecting the rate of hepatic androstenone metabolism which in turn may influence the level of hepatic CYP2E1 expression and hence the rate of hepatic skatole metabolism.     

Analysis of the genetics of boar taint reveals both single SNPs and regional effects

Background

Boar taint is an offensive urine or faecal-like odour, affecting the smell and taste of cooked pork from some mature non-castrated male pigs. Androstenone and skatole in fat are the molecules responsible. In most pig production systems, males, which are not required for breeding, are castrated shortly after birth to reduce the risk of boar taint. There is evidence for genetic variation in the predisposition to boar taint.A genome-wide association study (GWAS) was performed to identify loci with effects on boar taint. Five hundred Danish Landrace boars with high levels of skatole in fat (>0.3 μg/g), were each matched with a litter mate with low levels of skatole and measured for androstenone. DNA from these 1,000 non-castrated boars was genotyped using the Illumina PorcineSNP60 Beadchip. After quality control, tests for SNPs associated with boar taint were performed on 938 phenotyped individuals and 44,648 SNPs. Empirical significance thresholds were set by permutation (100,000). For androstenone, a ‘regional heritability approach’ combining information from multiple SNPs was used to estimate the genetic variation attributable to individual autosomes.

Results

A highly significant association was found between variation in skatole levels and SNPs within the CYP2E1 gene on chromosome 14 (SSC14), which encodes an enzyme involved in degradation of skatole. Nominal significance was found for effects on skatole associated with 4 other SNPs including a region of SSC6 reported previously. Genome-wide significance was found for an association between SNPs on SSC5 and androstenone levels and nominal significance for associations with SNPs on SSC13 and SSC17. The regional analyses confirmed large effects on SSC5 for androstenone and suggest that SSC5 explains 23% of the genetic variation in androstenone. The autosomal heritability analyses also suggest that there is a large effect associated with androstenone on SSC2, not detected using GWAS.

Conclusions

Significant SNP associations were found for skatole on SSC14 and for androstenone on SSC5 in Landrace pigs. The study agrees with evidence that the CYP2E1 gene has effects on skatole breakdown in the liver. Autosomal heritability estimates can uncover clusters of smaller genetic effects that individually do not exceed the threshold for GWAS significance.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-424) contains supplementary material, which is available to authorized users.     

Gender-related differences in the formation of skatole metabolites by specific CYP450 in porcine hepatic S9 fractions

Higher accumulation of skatole in the fat of male pigs compared with female pigs might be due to gender-related differences in the rate of skatole degradation. In the present study, skatole metabolites and cytochrome P450 (CYP450) isoforms involved in skatole metabolism were for the first time investigated in hepatic S9 fractions from six male and four female pigs (crossbred Landrace×Yorkshire dams and Duroc boar). Surprisingly, the rates of production of major skatole metabolites were similar in male and female pigs. The most abundant metabolite of skatole was 3-hydroxy-3-methyloxindole (HMOI) followed by 3-methyloxindole and indole-3-carbinol in both male and female S9 fractions. Concentrations of formed HMOI and 3-methyloxindole did not differ between the genders (P=0.124 for HMOI, and P=0.575 for 3-methyloxindole). Indole-3-carbinol formation was higher in S9 fractions from the females compared with male pigs (P=0.0001). Enzyme kinetic parameters were similar for both genders (P>0.05). In both male and female pigs, ellipticine, diallyl sulphide (DAS) and quercetin inhibited HMOI formation, confirming the involvement of CYP1A1 and CYP2E1. The formation of 3-methyloxindole was reduced in the presence of the CYP2E1 inhibitor DAS, and formation of indole-3-carbinol was reduced in the presence of CYP1A1 and CYP2A19 inhibitors. We found only minor differences in skatole metabolism between male and female pigs, particularly the involvement of CYP2C and CYP3A in indole-3-carbinol formation in female but not in male pigs. This is a very essential finding, suggesting the involvement of larger number of CYP450 isoforms in female pigs. On the other hand, indole-3-carbinol is a minor skatole metabolite, and the physiological significance of CYP2C and CYP3A involvement in its formation in female pigs, but not in male pigs, needs to be elucidated. Our results, however, should be interpreted with caution because of the low number of animals and possibility of breed and age effects on skatole metabolism.     

Effect of faecal soiling on skatole and androstenone occurrence in organic entire male pigs

Production of entire male pigs could be a future strategy for organic pig production. However, production of entire males leads to increased risk of carcasses with elevated boar taint levels. It is hypothesized that skatole levels in pig meat are affected by faecal soiling and that organic housing facilities can increase the risk of pigs being heavily soiled. Therefore, the overall aim of this study was to investigate if increased pig and pen soiling increases skatole concentration in entire male pigs. In five herds, 1174 organic entire male pigs were reared in four batches across two seasons, summer and winter. Measurements of pig and pen soiling, as well as fat skatole and androstenone concentration and human nose sensory tests of fat odour, were performed. Skatole and androstenone concentrations varied greatly within and between herds with a 10% and 90% percentile for the overall population of 0.02 and 2.25 µg/g for skatole and 0.53 and 4.84 µg/g for androstenone. Human nose positive tests averaged 18.3% with great variation between herds and seasons. Pen soiling had significant effects on pig soiling. Moreover, outdoor pen soiling significantly affected skatole concentration in interactions with herd and season (P<0.001 and P=0.003) and affected human nose positive risk in interaction with herd (P=0.005). Soiling on indoor pen areas did not affect skatole levels and no effect on androstenone was found for any pen area. Soiling of pigs affected both skatole and androstenone levels, with the size of the head and abdomen body areas covered in manure showing significant positive effects on skatole concentration. No effect of density of the manure layer was found on either boar taint measure. Herd significantly affected both skatole and androstenone in fat as well as the human nose positive risk. The human nose test revealed no effect from pig soiling. A large variation in the different boar taint measures was found for both high and low scores of pen and pig soiling, and only a small difference in skatole and androstenone concentrations between the high and low soiling categories was found. Therefore, while increasing the hygiene management could be a strategy for reducing boar taint in production of organic entire male pigs, it should be emphasized that other factors would also need to be considered.     

Molecular cloning and functional analysis of porcine <Emphasis Type="Italic">SULT1A1</Emphasis> gene and its variant: a single mutation <Emphasis Type="Italic">SULT1A1</Emphasis> causes a significant decrease in sulfation activity

The characterization of the SULT1A1 gene and its variants should have an important impact on the efforts to develop genetic markers to select for low skatole in pigs. Raising intact male pigs would have a significant economic impact on the pork industry; however, the presence of skatole (a major cause of boar taint) in meat from male pigs would be highly objectionable to consumers. It has been shown that the phase II metabolism of skatole metabolites by phenol sulfotransferase is related to the clearance of skatole in the liver. The aim of this study was to isolate and characterize the SULT1A1 gene from pig liver, examine its expression, identify genetic polymorphisms, and study how a genetic variation in this enzyme translates into interindividual variation in skatole levels. We show here that a substitution of AG at base 546 of SULT1A1 causes a significant decrease in its sulfation activity and thus may be at least partially responsible for a higher level of skatole in pigs. Our findings provide an important basis toward the goal of making it possible to predict the sulfation status in pigs and the development of genetic markers for low skatole.     

Detection of quantitative trait loci for androstenone, skatole and boar taint in a cross between Large White and Meishan pigs

'Boar taint' is a strong perspiration-like, urine-like unpleasant odour given off upon heating or cooking of meat from some intact (uncastrated) male pigs. Data from the F(2) generation of a Large White (LW) x Meishan (MS) crossbred population were analysed to detect quantitative trait loci (QTL) for traits associated with boar taint. Fat samples from 178 intact male pigs slaughtered at 85 +/- 5 kg were analysed for the major contributors to boar taint (androstenone, indole and skatole). Fat and lean samples from cooked meat were scored for boar, abnormal and pork flavour and odour by a trained sensory panel (SP). A scan with 117 markers covering the whole genome was performed in the F(2) individuals, together with their F(1) parents and purebred grandparents. At the 5% chromosomal significance threshold (approximately equal to the genome-wide suggestive significance threshold), QTL were detected for the laboratory estimate of androstenone on chromosomes 2, 4, 6, 7 and 9. However, only on chromosome 6 were there QTL for boar flavour (BF) traits in the same or adjacent marker intervals as a QTL for the laboratory estimate of androstenone. On chromosome 14, QTL were detected for the laboratory estimates of indole and skatole, the SP score for skatole and the scores for BF in lean and BF in fat. In all five cases, the MS allele generally increased the estimate or score, compared with the LW allele, but it appeared that desirable and undesirable alleles were present in both breeds. This locus on chromosome 14 has considerable potential for use to reduce the incidence of boar taint, especially if further research can identify the causative polymorphism or strongly associated markers.     

Hydrolysable tannin-based diet rich in gallotannins has a minimal impact on pig performance but significantly reduces salivary and bulbourethral gland size

Tannins have long been considered ‘anti-nutritional’ factors in monogastric nutrition, shown to reduce feed intake and palatability. However, recent studies revealed that compared with condensed tannins, hydrolysable tannins (HT) appear to have far less impact on growth performance, but may be inhibitory to the total activity of caecal bacteria. This in turn could reduce microbial synthesis of skatole and indole in the hindgut of entire male pigs (EM). Thus, the objective of this study was to determine the impact of a group of dietary HT on growth performance, carcass traits and boar taint compounds of group housed EM. For the study, 36 Swiss Large White boars were assigned within litter to three treatment groups. Boars were offered ad libitum one of three finisher diets supplemented with 0 (C), 15 (T15) or 30 g/kg (T30) of HT from day 105 to 165 of age. Growth performance, carcass characteristics, boar taint compounds in the adipose tissue and cytochrome P450 (CYP) isoenzymes CYP2E1, CYP1A2 and CYP2A19 gene expression in the liver was assessed. Compared with C, feed efficiency but not daily gain and daily feed intake was lower (P<0.05) in T15 and T30 boars. Except for the percentage carcass weight loss during cooling, which tended (P<0.10) to be greater in T30 than C and T15, carcass characteristics were not affected by the diets. In line with the numerically lower androstenone level, bulbourethral and salivary glands of T30 boars were lighter (P<0.05) than of T15 with intermediate values for C. Indole level was lower (P<0.05) in the adipose tissue of T30 than C pigs with intermediate levels in T15. Skatole levels tended (P<0.10) to be lower in T30 and C than T15 pigs. Hepatic gene expression of CYP isoenzymes did not differ between-treatment groups, but was negatively correlated (P<0.05) with androstenone (CYP2E1 and CYP1A2), skatole (CYP2E1, CYP2A) and indole (CYP2A) level. In line with the numerically highest androstenone and skatole concentrations, boar taint odour but not flavour was detected by the panellists in loins from T15 compared with loins from C and T30 boars. These results provide evidence that HT affected metabolism of indolic compounds and androstenone and that they affected the development of accessory sex glands. However, the effects were too small to be detected by sensory evaluation.     

Olfactory evaluation of boar taint: effect of factors measured at slaughter and link with boar taint compounds

There is a commitment by the European pig sector to ban surgical castration of male piglets in the European Union in 2018. One alternative to castration is to raise entire male pigs, with an increased risk of boar taint. A field study was performed to: (1) evaluate inter- and intra-farm variation in boar taint prevalence, (2) investigate factors measured at slaughter influencing boar taint and (3) evaluate the relationship between sensorial scoring by a trained panel and the concentration of boar taint components. From 34 farms, neck fat samples were collected from all entire male pigs in at least two slaughter batches per farm (78 batches; 9167 animals). In addition to olfactory boar taint analysis, data were also collected on fresh skin lesions (score 0 to 3) at the slaughter line, slaughter weight, lean meat percentage, duration of transport, time spent in lairage, total delivery duration, day length, shortening of days and outdoor mean temperature. Using the hot iron method, neck fat samples were scored (eight-point scale) for boar taint. Average boar taint prevalence (score ≥3) was 5.6±2.5% and the mean difference between the maximum and minimum prevalence per farm was 4.3±3.2%. Androstenone (AND), skatole (SKA) and indole concentrations were measured for a subset (n=254) of the samples. According to binomial univariate mixed models, entire male pigs with a higher skin lesion score had higher odds of having boar taint (P=0.031), as did fatter entire male pigs (P<0.001). In the binomial multivariate mixed model lean meat percentage (P<0.001) and outdoor mean temperature (P=0.005) remained as only significant factors. Based on our results, we can conclude that these statistically significant at least partially influence the prevalence of boar taint. According to the binomial univariate mixed models SKA concentration in liquid fat seems a better predictor for boar taint than AND. There were no significant synergetic effects between boar taint compounds.     

Pig meat quality from entire males

This paper constitutes an updated review of the production and meat quality aspects of rearing entire male pigs. Since a major obstacle in rearing entire males is the incidence of boar taint, possible methods for detection are also summarised. Safe and fast methods for detection of boar taint would be valuable in avoiding complaints from consumers. Pig meat quality is determined by many aspects, among which odour and taste are the most important attributes. Odour may be negatively affected by the presence of a pheromonal steroid, androstenone, and a fermentation product of l-tryptophan, skatole. Male pigs are surgically castrated in many countries to minimise the risk of accumulation of high levels of androstenone and skatole. Raising entire male pigs is more profitable because they have superior production characteristics and improved meat quality due to leaner carcasses and higher protein content, as compared to castrated pigs. Furthermore, surgical castration is negative from an animal welfare point of view. In most studies, no differences in sensory quality have been found between lean meat from entire male pigs with low levels of androstenone and skatole and pork from castrates and females. The question that remains is: which substances are responsible for boar taint besides androstenone and skatole and whether they need to be considered? The threshold values used for androstenone and skatole might also be too high for highly sensitive persons. Recent research shows that a human odorant receptor, ORD7D4, is involved in sensitivity to androstenone. If the ORD7D4 genotypes of consumer and expert panels are known, this might facilitate consumer studies in the future. There is still a great need for rapid on/at-line detection methods in abattoirs for identifying carcasses with unacceptable levels of boar taint compounds. Several emerging rapid technologies with a potential for boar taint detection have been investigated. They represent various measurement principles such as chemical sensor arrays (electronic noses), mass-spectrometry fingerprinting, ultra-fast gas chromatography, gas-phase spectrometry and biosensors. An industrial detection method should allow 100% correct classification of both acceptable and not-acceptable samples with regard to boar taint sorting criteria. There are, however, still too high a percentage of false negatives ranging from 5% to 20%. In addition, these methods do not yet seem to fulfil the industrial specifications with regard to cost efficiency, simplicity and analysis time. There is still no dedicated measurement technology available for on/at-line detection of boar-tainted carcasses that measures both androstenone and skatole.     

Cytochrome P450IIE1 (CYP2E1) is induced by skatole and this induction is blocked by androstenone in isolated pig hepatocytes

Skatole, a derivative of tryptophan, is produced in the hind-gut of pigs and is metabolised via hepatic cytochrome P4502E1 (CYP2E1). Excessive accumulation of skatole together with androstenone, a metabolite of testosterone, in adipose tissue in some pigs is a major cause of 'boar taint' and is associated with defective expression of CYP2E1. This phenomenon is not understood because factors regulating CYP2E1 expression in pig liver have not yet been characterised. Therefore effects of skatole and androstenone on CYP2E1 expression were studied using isolated pig hepatocytes as a model system. Skatole induced CYP2E1 protein expression to the same degree as did acetone, a known CYP2E1 inducer. Induction by skatole was maximum between 20 and 28 h and a half-maximum effect was obtained at a skatole concentration of 0.2 mM. Induction of CYP2E1 by skatole was protein-synthesis dependent. Androstenone antagonised the effect of skatole on CYP2E1 expression but did not affect the CYP2E1 protein level when added alone. These results suggest that defective expression of CYP2E1 in some pigs is due to excessive concentrations of androstenone which prevent CYP2E1 induction by its substrate skatole. As a result, skatole metabolism is reduced and skatole is accumulated in adipose tissue.     

Weight and season affects androstenone and skatole occurrence in entire male pigs in organic pig production

To investigate the extent to which the level of androstenone and skatole decreases with a decrease in live weight and/or age at slaughter of entire male pigs produced under organic standards, 1174 entire male pigs were raised in parallel in five organic herds, distributed across four batches in summer and winter. The median androstenone level was high for organic entire male pigs (1.9 µg/g), but varied greatly both within and between herds. Median skatole level was 0.05 µg/g, also with a wide range both within and between herds. Decreasing live weight over the range of 110±15.6 kg s.d. was found to decrease androstenone as well as skatole concentration, however, with different patterns of association. Age did not have significant direct effect on either androstenone or skatole levels. Androstenone levels were higher during winter than summer (P<0.0001), but no difference in skatole was found between seasons. The study concludes that decreasing live weight at slaughter could be an applicable management tool to reduce risk of boar taint and the level of tainted carcasses for a future production of entire male pigs within the organic pig production system, although further studies are needed as great variation in boar taint was found also for low weight animals.     

A performance test for boar taint compounds in live boars

Genetically reducing boar taint using low-taint lines is considered the most sustainable and economic long-term alternative to surgical castration of male pigs. Owing to the high heritability of the main boar taint components (androstenone, skatole and indole), breeding is an excellent tool for reducing the number of tainted carcasses. To incorporate boar taint into breeding programmes, standardized performance testing is required. The objective of this study was to develop and formally present a performance test for the main boar taint compounds on live breeding candidates. First, a standardized performance test for boar taint was established. A biopsy device was developed to extract small tissue samples (200 to 300 mg) from breeding candidates. Quantification of boar taint components from these small samples using specialized chemical extraction methods proved accurate and repeatable (r = 0.938). Following establishment of the method, biopsy samples of 516 live boars (100 to 130 kg live weight) were collected in the second step. Various mixed linear models were tested for each boar taint compound; models were ranked in terms of their information content. Pedigree information of 2245 ancestors of biopsied animals was included, and genetic parameters were estimated using univariate and multivariate models. Androstenone (in μg/g liquid fat (LF): mean = 0.578, σ = 0.527), skatole (in μg/g LF: mean = 0.033, σ = 0.002) and indole (in μg/g LF: mean = 0.032, σ = 0.002) levels obtained by biopsy were plausible. Heritability estimates for androstenone calculated with univariate (0.453) and multivariate (0.452) analyses were comparable to those in the literature. Heritabilities for skatole (0.495) and indole (0.550) were higher than that for androstenone. Genetic and phenotypic correlations were similar to those published previously. Our results show that data on boar taint compounds from small adipose samples obtained by biopsy provide similar genetic parameters as that described in the literature for larger samples and are therefore a reliable performance test for boar taint in live breeding candidates.     

Efficacy of non-nutritive sorbent materials as intestinal-binding agents for the control of boar taint

In many countries, male pigs are castrated to prevent boar taint, but this practice raises concerns about animal welfare and reduces the production efficiency of pork. The objective of this study was to develop dietary manipulations to prevent boar taint. We evaluated the effectiveness of adding activated carbon (AC) or Tween-60 (Tween; polyoxyethylene sorbitan monostearate) to pig finishing diets to reduce levels of androstenone (AND) and skatole in plasma and fat of entire male pigs. Boars (159 ± 2 days of age at the start of the experiment) were fed diets supplemented with either 5% AC or 5% Tween for 28 days followed by either 14 or 28 days of recovery. Plasma samples were collected at experimental days 0, 7, 14, 21, 28, 42 and 56, and backfat biopsies were taken at experimental days 0, 28, 42 and 56. Feeding AC significantly (P < 0.05) reduced the levels of AND in plasma by day 28 compared to day 0 and by day 42 in fat compared to day 0. AC treatment also decreased levels of oestrone sulphate (E(1)S) in plasma by day 7 compared to day 0. Treatment with Tween significantly decreased (P < 0.05) the levels of plasma AND by day 28 from levels at day 0. Tween treatment did not significantly affect levels of fat AND or plasma E(1)S compared to day 0; however, fat AND levels decreased between days 28 and 42 following treatment with Tween (P < 0.05). Levels of plasma E(1)S, plasma AND and fat AND for control boars remained constant throughout the experiment. Skatole plasma concentrations were very low and did not vary significantly (P > 0.05) from day 0 for any treatment, but fat skatole levels decreased by day 42 in the Tween treatment group. Importantly, there was no difference in growth rate between the control and experimental groups. We conclude that adding AC or Tween to finishing diets for boars can reduce the levels of plasma and fat AND, but further work is needed to confirm the effects of these treatments on reducing fat skatole levels.     

The effect of a c.-8G>T polymorphism on the expression of cytochrome b5A and boar taint in pigs

The level of cytochrome b5A ( CYB5A ) in pig testis is correlated with boar taint from androstenone and an AF016388:c.-8G>T polymorphism in CYB5A has been linked with low androstenone levels in the fat of pigs. In this study, we developed a polymerase chain reaction-based assay to genotype 1242 boars from eight lines for the c.-8G>T SNP. The c.-8T allele was found in all eight lines at a frequency ranging from 1.8% to 20.3% with an overall frequency of 8.6%. Significant deviations from Hardy–Weinberg equilibrium were found in the Hampshire, Landrace and Yorkshire breeds. The homozygous mutant c.-8TT occurred infrequently and was not found in some lines, but was consistently associated with low androstenone levels in fat. Both CYB5A mRNA and CYB5A protein levels were decreased in the c.-8TT genotype in a subset of Yorkshire boars, suggesting that low levels of CYB5A protein in the c.-8TT mutant were not due to inefficient translation of CYB5A mRNA. There were significant but modest marker effects on fat androstenone levels in Landrace, Yorkshire and a Large White/Duroc cross and fat skatole in Duroc and Sire Line breeds. There was no effect of CYB5A genotype on bulbourethral gland length, suggesting that this SNP will not affect reproductive traits. We conclude that the c.-8G>T SNP in the CYB5A gene has a significant but modest effect on boar taint in male pigs and could be useful in some breeds as part of a panel of SNP markers in a marker-assisted selection programme to produce low boar taint pigs.