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The accumulation of skatole in fat tissue is one of the predominant factors, causing boar taint. The present study was aimed to understand the mechanism whereby active immunization against GnRH (immunocastration) eliminates skatole in boars. Thirty-six boars were assigned within litter into three groups (n = 12): control, surgically castrated, or immunized against GnRH at 10 wk of age (with a booster 8 wk later). Faecal and blood samples (for skatole and skatole-regulatory hormone profiles) were collected at 4-wk intervals until boars were slaughtered (26 weeks). Immunocastration reduced (P < 0.05) serum levels of androstenone, 17β-estradiol and IGF1 especially after the booster immunization, and down-regulated (P < 0.05) mRNA expressions of both IGF1 and IGF1receptor (IGF1R) in mucosa of ileum as well as colon at slaughter. Compared to intact controls, immunocastration substantially decreased (P < 0.05) faecal skatole contents subsequent to the decrease of serum IGF1 levels, which persisted in boars after surgical castration. In parallel with the decreased formation of skatole in the intestine, levels of skatole in serum and then in fat tissue were also decreased (P < 0.05). On the other hand, deprivation of testicular steroids, especially androstenone and 17β-estradiol accelerated skatole degradation metabolism in the liver by increasing (P < 0.05) hepatic CYP2E1, CYP2A, CYP2C49 and CYB5A expressions. Collectively, our results suggested that immunocastration decreased skatole formation in the intestine and meanwhile accelerated skatole degradation metabolism in the liver, resultantly eliminating skatole accumulation in male pigs. Decreased intestinal skatole formation by immunocastration appeared to be associated with the attenuated actions of IGF1 on the turnover of both ileal and colon mucosa., (Copyright © 2019 Elsevier Inc. All rights reserved.)

A high performance liquid chromatographic (HPLC) method with fluorescence detection (FD) was established to simultaneously determine plasma indole and skatole. Plasma samples were pretreated by liquid-liquid extraction. Chromatographic separation was accomplished on a Shim-Pack VP-ODS column (150 mm×4. 6 mm, 4.6 μ m) using an isocratic mixture of 15 mmol/L sodium dihydrogen phosphate solution and methanol (40:60, v/v). The fluorescence excitation and emission wavelengths were 274 nm and 340 nm, respectively. The linear ranges were 2.22-88.89 μ g/L for indole and 1.11-44.44 μ g/L for skatole. The detection limits were 0.11 μ g/L and 0.06 μ g/L for indole and skatole, respectively. The recoveries were in the range of 95.5%-112.3% with the relative standard deviations less than 6.8%. The method was successfully applied to the analysis of plasma from healthy pregnant women ( n =46) and pregnant women with hepatitis B virus (HBV) infection ( n =29). The results showed that plasma indole and skatole levels were significantly different between two groups. In pregnant women with HBV infection, the concentrations of indolic compounds were positively associated with transaminase levels.

Skatole leads to off-odor in pork and is influenced by several factors such as sex and management conditions of pigs, but the causal relationships have not yet been clarified. In the present study, physiological skatole concentrations along the carcass were monitored and the transdermal diffusion of skatole was experimentally studied with skatole-spiked feces. Additionally, the impact of different biopsy techniques on skatole in fat and blood was studied. Monitoring of skatole along the carcass revealed higher skatole concentrations in the belly than in dorsal cuts. Topical application of spiked feces increased skatole in fat strictly at the application site. In contrast to punch biopsies, surgical biopsies significantly affected skatole and cortisol levels in blood, but not in fat. We conclude that biopsies for skatole measurements should be taken without anesthesia from the dorsal side of the animals. Fecal contaminations on the ventral side are not likely to influence overall concentrations., (Copyright © 2015. Published by Elsevier Ltd.)

Boar taint is an off-odour occurring while heating meat or fat from boars. A method detecting the three compounds (androstenone, skatole and indole) simultaneously in blood would offer substantial advantages since it would allow monitoring the impact of rearing strategies. Therefore, a UHPLC-HR-Orbitrap-MS analysis method is optimized and validated for the quantification of these compounds in plasma or serum. Sample pre-treatment involved an extraction with diethylether followed by a centrifugal filtration (30 kDa). Limits of detection and quantification varied between 0.5 and 1 μg L(-1) and 2 and 3 μg L(-1) for the three compounds, respectively. Besides, an excellent repeatability (RSD < 7.6%), within-laboratory reproducibility (RSD<10.5%), recovery (87-97%) and linearity (R(2)>0.99) were recorded. Correlations between serum/plasma and fat levels of the boar taint compounds were positive for skatole (r(serum) = 0.39 and r(plasma) = 0.84) and androstenone (r(serum) = 0.73-0.78 and r(plasma) = 0.32-0.80)., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Societal pressure to ban surgical castration of male piglets is rising due to animal welfare concerns, thus other methods to prevent boar taint need to be explored. Genetic selection against boar taint appears to be a long-term sustainable alternative. However, as boar taint is linked to reproductive hormones, it is important to consider possible negative side effects such as delayed sexual maturity or changes in behaviour. We reported earlier that the melanocortin-4 receptor (MC4R) marker can be used to reduce boar taint levels in fat of boars. The objective of this study was to evaluate whether MC4R marker-assisted selection for lower boar taint prevalence affects plasma levels of boar taint compounds and testosterone; sexual maturity; behaviour; skin lesions; and lameness in boars and gilts. Using an intervention study with a 2×2 design, 264 boars and gilts differing on position 893 of the MC4R gene (AA v. GG) were compared. The MC4R polymorphism did not affect the plasma concentration of either androstenone or testosterone at different time points, whereas the concentration of skatole was significantly lower (P=0.003) and the concentration of indole tended to be lower (P=0.074) in GG compared with AA boars. A higher percentage of gilts of the GG genotype were in puberty at slaughter age compared with AA gilts (P<0.001). The age of the boars at sexual maturity (as indicated by the first positive preputial smear test) did not differ between AA and GG boars. In contrast, weight of GG boars at sexual maturity tended to be lower (P=0.065). During the period from 6 weeks of age to slaughter, boars and gilts of the GG genotype showed more playing behaviour (P=0.015) and less passive and feeding behaviour (P=0.003). They showed more skin lesions on their back and caudal area (P=0.022), and tended to show more skin lesions on their head and anterior area (P=0.093) compared with AA animals. In conclusion, the polymorphism in the MC4R gene can be used as a marker without negative effects on reproduction characteristics in boars and gilts. Genetic selection towards a lower prevalence of boar taint will lead to more active pigs with more skin lesions. Management strategies may therefore be necessary to reduce skin lesions in the selected animals.

Rearing entire pigs may lead to meat quality and welfare problems in relation to pubertal development. A better knowledge of the sources of variation of pubertal development, behaviour and boar taint is needed before generalizing entire male pigs. From 84 days of age, entire male pigs were reared in groups of 10 either in a conventional (C, 1 m²/animal, slatted floor) or an enriched (E, 2.5 m²/animal, straw bedding, outdoor run) housing during spring or autumn and fed ad libitum (n=10/housing/season). Mounting behaviour was observed for 3 h during the third (M3), fourth (M4) and fifth (M5) months of age. The total number of skin lesions was counted on both sides of the pigs 1 day before the behavioural recordings. The time spent in the outdoor run was also recorded during 3 days per month. The animals were slaughtered at 161±1 days of age (122±9 kg live weight). Blood samples were collected at 89 (M3), 119 (M4) and 152 (M5) days of age and at slaughter for the testosterone and oestradiol measurements. The testes were collected at slaughter, freed from the surrounding tissues and weighed. The fat samples were collected for the androstenone and skatole concentration measurement. Plasma testosterone and oestradiol-17β (oestradiol), fat androstenone and skatole and weight of the testes did not differ between the housing systems. Plasma testosterone (8.3 v. 3.9 nmol/l, P<0.05) and oestradiol (12.0 v. 9.2 pmol/l, P<0.1) at M3, fat skatole (0.124 v. 0.043, P<0.03) and weight of the testes (587 v. 512 g, P<0.05) were higher in the autumn than in the spring trial, suggesting that the pubertal development was accelerated. The number of received mounting behaviours was slightly higher in the autumn (P=0.08) trial and was markedly higher in the E than in the C environment (P<0.003). Skin lesions were more numerous in the C than in the E housing at M4 and M5 and in the spring than in the autumn trial at M3 and M4 (P<0.05). Fat androstenone and the number of performed mounting behaviours were significantly correlated between each other and with numerous indicators of the pubertal development (P<0.05). The number of skin lesions was correlated with plasma testosterone and live weight (P<0.05). Overall, this study suggests the effect of season on sexual development, the effect of the housing system on behaviour, and demonstrates the links between sexual hormones, behaviour and boar taint.

The aim of this study was to investigate the effect of a vaccine against gonadotropin-releasing hormone (GnRH), Improvac(®) (Pfizer Ltd), administered at a pre- or early pubertal stage on boar taint, hormonal status and reproductive organs. Crossbred male pigs (Swedish Yorkshire dams×Swedish Landrace sires or Swedish Yorkshire sires, n=192) were at birth randomly allocated to four groups: one group of pigs surgically castrated without anaesthesia before age 1 week, a second group of early vaccinated pigs given Improvac at ages 10 and 14 weeks, a third group of standard vaccinated pigs given Improvac at ages 16 and 20 weeks, and a fourth group of entire male pigs. Following the second vaccine injection, antibody titres increased rapidly, accompanied by a rapid decrease in testosterone and a slower decrease of skatole in plasma to the same low levels as for surgically castrated pigs. At slaughter, the levels of androstenone and skatole in adipose tissue were low in surgically castrated and vaccinated pigs, whereas entire male pigs had elevated levels (p<0.001). Similarly, oestradiol was at low levels for surgically castrated and vaccinated pigs, whereas entire male pigs had elevated levels (p<0.001). IGF-1 was lowest for surgically castrated pigs and highest for entire male pigs, with vaccinated pigs at an intermediate level (p<0.001). At slaughter, reproductive organs were small in pigs vaccinated with Improvac, and smaller in pigs vaccinated early (p<0.001). Under our experimental conditions, early vaccination with Improvac can be used as an alternative to the recommended schedule with maintained control of boar taint and testicular secretory activity., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

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., (Copyright © 2010 The American Meat Science Association. Published by Elsevier Ltd. All rights reserved.)

The concentrations of the boar taint compounds androstenone and skatole in plasma and fat, together with those of testosterone in plasma, were investigated in pubertal purebred Duroc and Landrace boars following stimulation with human chorionic gonadotrophin (hCG). Higher initial levels of androstenone and testosterone were found in Duroc than Landrace boars. Duroc boars, which were approximately ten days older than the Landrace boars, also showed a more advanced stage of spermatogenesis than Landrace boars. While Landrace boars had the highest skatole levels. Following stimulation with hCG the relative increases in testosterone, androstenone, and skatole concentrations were highest in Landrace boars. The level of androstenone in fat three days after hCG stimulation exceeded 1 microg/g fat in all stimulated boars. The decreases in plasma levels of androstenone and testosterone on Days 2 and 3 after hCG stimulation were more pronounced in Landrace than Duroc boars. However, unlike the plasma androstenone and testosterone levels, the plasma concentrations of skatole did not decrease on Days 2 and 3 following stimulation, but remained elevated on Day 3. These results indicate that the lower levels of testicular steroids in Landrace boars compared with Duroc boars was not due to a lower production capacity, but more likely to a faster disappearance of steroids in Landrace boars. In the present study, age, live weight, and testicular development did not significantly contribute to the variation in fat androstenone. The present data and previous reports on candidate genes related to androstenone biosynthesis and metabolism suggests that future selection against factors associated with boar taint remains a possible solution for the problem of boar taint in the swine industry., ((c) 2010 Elsevier Inc. All rights reserved.)

Background: Boar taint is an unpleasant odour and flavour of the meat from some uncastrated male pigs primarily caused by elevated levels of androstenone and skatole in adipose tissue. Androstenone is produced in the same biochemical pathway as testosterone and estrogens, which represents a particular challenge when selecting against high levels of androstenone in the breeding programme, without simultaneously decreasing levels of other steroids. Detection of single nucleotide polymorphisms (SNPs) associated with compounds affecting boar taint is important both for gaining a better understanding of the complex regulation of the trait and for the purpose of identifying markers that can be used to improve the gain of breeding. The beneficial SNPs to be used in breeding would have the combinational effects of reducing levels of boar taint without affecting fertility of the animals. The aim of this study was to detect SNPs in boar taint candidate genes and to perform association studies for both single SNPs and haplotypes with levels of boar taint compounds and phenotypes related to reproduction., Results: An association study involving 275 SNPs in 121 genes and compounds related to boar taint and reproduction were carried out in Duroc and Norwegian Landrace boars. Phenotypes investigated were levels of androstenone, skatole and indole in adipose tissue, levels of androstenone, testosterone, estrone sulphate and 17beta-estradiol in plasma, and length of bulbo urethralis gland. The SNPs were genotyped in more than 2800 individuals and several SNPs were found to be significantly (LRT > 5.4) associated with the different phenotypes. Genes with significant SNPs in either of the traits investigated include cytochrome P450 members CYP2E1, CYP21, CYP2D6 and CYP2C49, steroid 5alpha-reductase SRD5A2, nuclear receptor NGFIB, catenin CTNND1, BRCA1 associated protein BAP1 and hyaluronoglucosaminidase HYAL2. Haplotype analysis provided additional evidence for an effect of CYP2E1 on levels of skatole and indole, and for BAP1, HYAL2 and SRD5A2 on levels of androstenone., Conclusion: The findings in this study indicate that polymorphisms in CYP2E1, CYP21, CYP2D6, CYP2C49, NGFIB and CTNND1 might be used to reduce levels of boar taint without affecting levels of testosterone, estrone sulphate, 17beta-estradiol or length of bulbo urethralis gland.

The objective of this study was to evaluate the long-term effect of a gonadotropin-releasing hormone (GnRH) vaccine, Improvac (Pfizer Ltd.), on the levels of GnRH antibodies, testosterone, estrone sulphate (E1S) and androstenone, as well as skatole and indole in male pigs. Additionally, the long-term effect of immunocastration on social and sexual behaviour was studied. Male pigs were assigned to two treatment groups: a treatment group given two doses of Improvac (n=12) and a control group of entire male pigs (n=12). The pigs were kept either 16 or 22 weeks after vaccination. Blood samples were collected five or six times; prior to both first and second vaccination, then three or four times during the 16 or 22 week period after second vaccination. Immunocastration significantly reduced levels of testosterone and E1S in plasma, and levels of androstenone in fat (P<0.001 for all). Skatole and indole levels in plasma and fat were also lower in immunocastrated pigs than in entire male pigs. These effects lasted up to 22 weeks after the second vaccination. Testis weight and bulbourethral gland length were lower in immunocastrated pigs at slaughter and these pigs showed less social, manipulating and aggressive behaviour than entire male pigs. The immunocastrated pigs remained sexually inactive throughout the study. Our study represents a further step in the evaluation of the effectiveness of Improvac as an alternative to surgical castration of entire male pigs. It shows that Improvac may have an extended effect compared with that currently implied by the directions for use.

The accumulation of skatole in boars to concentrations resulting in carcass taint has been associated with elevated concentrations of steroid hormones in plasma. Studying boar taint in vivo has been challenging because steroid hormones are highly variable between individual boars. However, a peak in steroid hormones occurs between 2 and 4 wk postpartum; therefore, skatole production was investigated in the prepubescent pig. Plasma concentrations of estrone sulphate, dehydroepiandrosterone sulphate, and testosterone peaked between 2 and 4 wk postpartum in intact male pigs, whereas plasma concentrations of these steroid hormones remained low or undetectable in gilts and barrows. However, plasma skatole concentration peaked in all 3 groups of animals between 2 and 3 wk postweaning. The effects of weaning time, intestinal cell turnover, and diet on tissue skatole concentrations were then investigated. Intact male piglets were weaned at 14, 21, 28, or 35 d of age. Plasma skatole concentrations were measured weekly for a period of 63 d and peaked at 17 +/- 1, 14 +/- 1, 13 +/- 1, and 10 +/- 2 d postweaning, respectively. Intestinal cell turnover, as evaluated by villous height:crypt depth ratio, was not correlated with skatole concentrations in cecal contents, suggesting that cellular debris did not constitute a gross source of tryptophan for hindgut fermentation. The inclusion of 10% chicory inulin to piglet diets suppressed the postweaning increase in plasma skatole. Cecal skatole concentrations were also 3.3-fold lower in inulin-supplemented piglets compared with controls. The rise in plasma skatole in the prepubescent intact male pig was not associated with increased steroidogenesis but is likely due to the postweaning adaptation of the intestinal flora to an abrupt dietary change.

Relationship between free oestrone and boar taint compounds in adipose tissue were studied in two groups of entire male pigs of different breeds. Group A consisted of 33 entire crossbred male pigs (dam Yorkshire and sire backcross Yorkshire x Wild Boar, generation seven). Group B consisted of 194 entire male pigs of crossbreeds between Swedish Hampshire (H) and Finnish Landrace (L), LH x H, H x LH, LH x LH (dam x sire). The measurements of free oestrone in adipose tissue were performed with a new method developed and validated in our laboratories. The standard curve was linear for concentrations of free oestrone ranging from 0.13 to 5.10 ng/g. The method exhibited parallelism of results between serial dilutions and a mean recovery of 97 +/- 13.7%. Intra-assay variations for samples with concentrations of free oestrone from 0.67 to 2.08 ng/g were from 9.23 to 11.94%. Inter-assay variations for the samples with concentrations of free oestrone from 0.89 to 2.96 ng/g were from 3.78 to 10.11%. The levels of free oestrone in fat from group A were well correlated with fat levels of androstenone (r = 0.66; p < 0.001) and levels of oestrone sulphate in peripheral plasma collected at the same time as the fat (r = 0.74, p < 0.001). The levels of free oestrone in fat from group B were significantly correlated to fat levels of androstenone (r = 0.68, p < 0.001) and skatole (r = 0.29, p < 0.001). In group B, age-related differences in fat levels of free oestrone, androstenone and skatole were studied. Free oestrone and skatole levels increased simultaneously at the age of 22 week (p < 0.05 for both), and androstenone levels increased at the age of 26 week (p < 0.05). It was suggested that the levels of free oestrone in adipose tissue might be used for the evaluation of hormonal status of male pigs as an alternative to plasma levels of testicular hormones. The levels of free oestrone might be involved in the regulation of skatole levels in fat as indicated by both the simultaneous increases in skatole and free oestrone levels in fat and positive correlation between skatole and free oestrone.

This study describes the age-related variation in boar taint compounds, skatole and androstenone, and testosterone, oestradiol-17 beta (E17 beta), oestrone sulphate (ES), dehydroepiandrosterone sulphate (DHEAS), triiodothyronine (T(3)) and insulin-like growth factor-1 (IGF-1) in six boars. Three pairs of littermates of crossbred entire male pigs (from three Yorkshire x Duroc dams and one Hampshire sire) were included. Blood samples were taken at the age of 9-15 weeks and thereafter at weekly intervals from the age of 20-32 weeks. Plasma concentrations of skatole, androstenone, testosterone, E17 beta, ES, DHEAS, T(3) and IGF-1 were measured. We found that skatole levels in boars increased at the age around puberty after an increase in the levels of testicular steroids. Levels of skatole were not associated with the levels of sex steroids, T(3) and IGF-1. However, the increased level of testicular steroids is probably the underlying factor needed for high skatole levels to occur although the specific mechanism leading to increased skatole levels remains unknown.

Objective: To evaluate the effect of feeding aspirin and supplemental vitamin E on growth performance, lung lesions, plasma concentrations of 3-methylindole (3MI), and 3-methyleneindolenine (3MEIN)-adduct concentrations in blood and pulmonary tissues of feedlot cattle., Animals: 256 crossbred steers; 64 cattle were used in experiment 1 and 192 cattle were used in experiment 2., Procedures: A 2 X 2 factorial design was used for each experiment. Treatment factors were aspirin (0 or 3 g daily) and vitamin E (200 or 1,500 IU daily). Steers were housed in pens (8 steers/pen). Steers were slaughtered on days 59 and 138 for experiments 1 and 2, respectively. Lungs were grossly evaluated. 3MEIN-adduct concentrations were determined, and blood and pulmonary tissues., Results: Treatment was not associated with improvement or adverse effects on weight gain, dry-matter intake, or feed efficiency in experiment 2. In experiment 1, 36 of 63 (57.1%) steers had lung lesions. Lesions were not associated with treatment or concentrations of 3MI and 3MEIN-adduct. Plasma 3MI concentration and concentrations of 3MEIN-adduct in blood and pulmonary tissues were 3.11 microg/mL, 0.51 U/microg of protein, and 0.49 U/microg of protein, respectively. Aspirin was associated with increased blood concentrations of 3MEIN-adduct for diets that did not contain supplemental vitamin E., Conclusions and Clinical Relevance: Differences n performance of feedlot steers were not associated with treatment diet. It is possible that concurrent exposure of feedlot cattle to other factors typically associated with development of respiratory tract disease would affect these findings.

Objective: To describe time-dependent changes in plasma concentrations of 3-methylindole (3MI) and blood concentrations of 3-methyleneindolenine (3MEIN)-adduct in feedlot cattle., Animals: 64 yearling steers., Procedures: Steers were assigned to 2 groups (32 steers/group). During the first 8 weeks, blood samples were collected from group 1 before the morning ration was fed, whereas samples from group 2 were collected 2 to 3 hours after the ration was fed. Blood samples were collected from all steers approximately 4 times/wk for 3 weeks and 3 times/wk for the subsequent 5 weeks. Samples were collected at the same time for all steers for an additional 10 weeks. Plasma samples were analyzed for 3MI concentrations. Blood samples collected from cattle in group 2 during the first 8 weeks were analyzed for 3MEIN-adduct concentrations., Results: Mean blood concentration of 3MEIN-adduct increased to a maximum value on day 33 (0.80 U/microg of protein) and then decreased to a minimum on day 54 0.40 U/microg of protein). Plasma 3MI concentrations initially decreased and remained low until after day 54. Group-1 cattle had lower plasma 3MI concentrations, compared with concentrations for group-2 cattle. Blood 3MEIN-adduct concentrations and plasma 3MI concentrations were not associated with deleterious effects on weight gains., Conclusions and Clinical Relevance: Blood 3MEIN-adduct concentrations peaked during the period of greatest risk for development of bovine respiratory disease complex. Conversely, plasma 3MI concentrations decreased during the same period. Animal-to-animal variation in metabolic capacity to convert 3MI to 3MEIN may be of more importance than differences in plasma 3MI concentration.

Field data were collected over 2 consecutive years to characterize acute interstitial pneumonia (AIP) in feedyard cattle. Thirty-eight cattle with clinical symptoms of AIP were examined following emergency slaughter; 31 (all heifers) were confirmed to have AIP on the basis of gross and histological lung pathology. The 7 without AIP, plus 17 asymptomatic penmates, were used as contemporary controls. Plasma concentrations of 3-methylindole (3MI) metabolites were higher (P < 0.001) in heifers afflicted with AIP than in the control animals, and concentrations of 3MI mercapturates in the urine were lower (P < 0.007) in affected heifers. Concentrations of 3MI adducts in lung tissue and in microsomal protein did not differ (P > 0.05) between the 2 groups, and 3MI was not detected in ruminal fluid from either group. Total ruminal bacterial numbers and populations of lactobacilli and protozoa were similar (P > 0.05) between the AIP-positive and unafflicted groups, but fewer (P < 0.05) cellulolytic bacteria were present in the positive group. Bovine respiratory syncytial virus antigen was not found in lung tissue from any of the heifers confirmed to have AIP. To our knowledge, this study is the first to implicate 3MI metabolites as having a role in feedyard AIP. Further research is required to determine the factors responsible for the elevation in 3MI adducts in plasma and urine of feedyard cattle afflicted with AIP.

Objective: To evaluate the potential synergy between bovine respiratory syncytial virus (BRSV) and 3-methylindole (3MI) in inducing respiratory disease in cattle., Animals: 20 mixed-breed beef calves., Procedure: A 2 X 2 factorial design was used, with random assignment to the following 4 treatment groups: unchallenged control, BRSV challenge exposure (5 X 10(4) TCID50 by aerosolization and 5.5 X 10(5) TCID50 by intratracheal inoculation), 3MI challenge exposure (0.1 g/kg of body weight, PO), and combined BRSV-3MI challenge exposure. Clinical examinations were performed daily. Serum 3MI concentrations, WBC counts, PCV, total plasma protein, and fibrinogen concentrations were determined throughout the experiment. Surviving cattle were euthanatized 7 days after challenge exposure. Pulmonary lesions were evaluated at postmortem examination., Results: Clinical respiratory disease was more acute and severe in cattle in the BRSV-3MI challenge-exposure group than in cattle in the other groups. All 5 cattle in this group and 3 of 5 cattle treated with 3MI alone died or were euthanatized prior to termination of the experiment. Mean lung displacement volume was greatest in the BRSV-3MI challenge-exposure group. Gross and histologic examination revealed that pulmonary lesions were also most severe for cattle in this group., Conclusions and Clinical Relevance: Feedlot cattle are commonly infected with BRSV, and 3MI is produced by microflora in the rumen of all cattle. Our results suggest that there is a synergy between BRSV and 3MI. Thus, controlling combined exposure may be important in preventing respiratory disease in feedlot cattle.

The relationships between levels of skatole and androstenone in fat, plasma levels of skatole metabolites, plasma testosterone and estrone sulfate, and levels of cytochrome P450IIE1 in liver were investigated in Swedish Yorkshire pigs and F4 Wild Pig crosses. Levels of skatole, the skatole metabolite MIII, and androstenone were higher in males from the Wild Pig crosses than in the Yorkshire pigs. Plasma levels of the skatole metabolite MII and the ratio between MII and MIII were higher in female Yorkshire pigs than in male pigs. Skatole was correlated with androstenone, cytochrome P450IIE1, and estrone sulfate in the Wild Pig crosses but not in Yorkshire pigs. However, plasma levels of skatole metabolites were not correlated to levels of cytochrome P450IIE1 in liver in either group on animals. Pigs with high levels of cytochrome P450IIE1 in liver had low levels of skatole in fat, and pigs with low levels of cytochrome P450IIE1 in liver had low and high skatole levels. Using stepwise regression analysis, only levels of cytochrome P450IIE1 in liver significantly explained the variation in skatole levels in the Wild Pig crosses. Our results support the hypothesis that low levels of cytochrome P450IIE1 in liver may result in high levels of skatole in backfat of uncastrated male pigs due to decreased metabolism and clearance of skatole. However, these pigs cannot be identified based on plasma levels of skatole metabolites MII and MIII.

Five Hereford cows were given an intraruminal dose of L-tryptophan (0.35 g/kg of body weight), and 2 cows were used as controls. Of the 5 treated cows, 3 developed clinical signs of interstitial pul monary edema, and emphysema and severe pulmonary lesions were seen at necropsy after 96 hours. Another cow developed moderate clinical signs and pulmonary lesions, and the remaining cow had few clinical signs and mild pulmonary lesions. The severity of clinical signs in each cow was related to the severity of pulmonary lesions at necropsy. The 3-methylindole (3MI) was present in ruminal fluid and plasma within 6 hours after administration of tryptophan, and the concentrations increased to 3.0 and 9.0 mug/ml within 12 to 24 hours. Severity of pulmonary lesions was related to maximal concentration and duration of 3MI in the plasma. At necropsy, gross lesions were characterized by diffuse, pulmonary edema and interstital emphysema; and the lungs were dark red, firm, and heavier than normal. Predominant microscopic changes included accumulation of proteinaceous residue, hypertrophy and hyperplasia of alveolar lining epithelium, thickening of alveolar septums, and emphysematous thickening of interstitial tissues. These changes were similar to previously reported 3MI-induced pulmonary lesions. The presence of 3MI in ruminal fluid and plasma after administration of tryptophan and the relationship between concentration of 3MI and severity of clinical signs indicate that 3MI is the principal metabolite of ruminal fermentation which leads to the development of acute pulmonary edema and emphysema in cattle given tryptophan.

To study the role of cytochrome P-450-dependent mixed function oxidase reactions in equine 3-methylindole (3MI) toxicosis, ponies were given 20 mg of phenobarbital/kg of body weight at 72, 60, 48, 36, and 24 hours before 100 mg of oral 3MI/kg to induce cytochrome P-450 or no treatment (controls). Maximal 3MI plasma concentration was decreased and clearance was faster in phenobarbital-treated ponies. Plasma 3MI was still detectable 12 and 36 hours after dosing in phenobarbital-treated and control ponies, respectively. Phenobarbital treatment induced a distribution phase with transition from a 1-compartment to a 2-compartment extravascular model. Bronchiolitis occurred in all ponies 72 hours after 3MI, but was more severe in those treated with phenobarbital. Appearance of a distribution phase, increased total body clearance, and more severe bronchiolitis in phenobarbital-treated ponies indicated that mixed function oxidases are involved in metabolism and conversion of 3MI to a toxic metabolite.

Intraruminal and intravenous administration of 3-methylindole (3MI; skatole) caused interstitial pulmonary edema and emphysema in cattle. In 3 adult heifers given the intraruminal dose of 0.2 g of 3 MI per kilogram of body weight, clinical signs of respiratory disease appeared between 6 and 12 hours after dosing, and death due to pulmonary edema and emphysema occurred at 33, 69, and 72 hours. The mean plasma concentration of 3MI became maximal (18.5 mug/ml) at 3 hours and then decreased to low concentrations by 48 hours. In 2 heifers given an intraruminal dose of 0.1 g of 3MI/kg, clinical signs developed, but they did not die during the 96-hour experiment. The mean plasma concentration of 3 MI became maximal (16.8 mug/ml) at 3 hours and decreased to 1.6 and 0.4 mug/ml at 12 and 36 hours, respectively. At necropsy of the heifers, the lung were large, firm, dark red, and heavier than normal. Diffuse pulmonary edema was the predominant change in cattle which died early, and interstitial emphysema was more severe at later stages of the disease. During the early stages, alveoli were overdistended, and a few more ruptured. Most alveolar spaces were filled with proteinaceous residue, but the alveolar septums were smooth and of normal thickness. At later stages, proliferation of alveolar cells was observed, and alveolar septums were thickened. In 3 cows given 0.06 g of 3MI/kg by jugular infusion, clinical signs appeared in all cows, and 1 cow died of pulmonary edema and emphysema 56 hours after the infusion was started. Severe pulmonary lesions seen in all of the cows given a 3MI infusion were similar to those in the cows given an intraruminal dose of 3MI. The mean plasma concentration of 3MI increased to 10.7 mug/ml at 9 hours after starting the infusion and decreased to 0.5 mug/ml at 18 hours. The results indicate that 3MI, a product of ruminal tryptophan fermentation, can cause pulmonary edema and interstitial emphysema in cattle and support the hypothesis that 3MI is the causative agent in tryptophan-induced pulmonary disease.

The effects of intraruminal administration of 3-methylindole (3MI; skatole) were determined in goats. The 3MI was given to 4 goats at the dose level of 0.3 g/kg of body weight, to 2 goats at 0.2 g/kg, and to 2 goats at 0.1 g/kg; 3 nontreated goats were used as controls. Clinical signs of acute progressive respiratory tract disease were seen in all treated goats. Goats given the largest dose of 3MI (0.3 g/kg) died between 5 and 11 hours after treatment; those given smaller doses (0.2 and 0.1 g/kg) died between 79 and 92 hours. Increased plasma concentrations of 3MI were detected in goats give 0.1 or 0.2 g/kg within 3 hours after administration. By 24 and 36 hours, the concentrations of 3MI in the plasma decreased to low or nondetectable amounts and remained low for the duration of the experiment. Clinical signs of respiratory distress in the goats progressed after 3MI had been cleared from the plasma. Diffuse pulmonary edema and hydrothorax were extensive in goats which died early in the course of the experimentally induced disease. In goats which died at later stages, the lungs were firm and had less watery transudate. Temporal variations in the nature of pulmonic changes were even more obvious by microscopic examination. Diffuse pulmonary edema was the predominant early change. Small foci of emphysema were apparently caused by overdistention of some clusters of alveoli. Marked septal thickening and proliferation of alveolar cells were the prominent changes in goats which died between 79 and 92 hours after treatment. Incubation of L-tryptophan with caprine ruminal fluid resulted in formation of indoleacetic acid, indole, and 3MI. Similar incubations did not convert indoleacetic acid to 3MI. Control incubations showed 3MI as a fermentation metabolite, indicating it exists in caprine ruminal fluid in vivo. Results demonstrated that goats are susceptible to intraruminal administration of 3MI. The transitory appearance of 3MI in the plasma associated with progressive respiratory tract disease was similar to observations in cattle give 3MI. Clinical signs and lesions seen at necropsy were qualitatively similar to those reported in cattle given tryptophan and indoleacetic acid.

A modified gas liquid chromatography method has been developed to analyse the concentrations of indole and 3-methylindole in samples of plasma and rumen fluid from four cows with fog fever together with four control animals and from two cattle given L-tryptophan orally. Satisfactory recoveries of the metabolites were obtained in test samples examined by the gas liquid chromatography procedure and increased levels were found in rumen fluid after tryptophan administration. Absence of abnormal concentrations of the toxic metabolite 3-methylindole in samples from field cases of fog fever was not considered to rule out the metabolite as a possible cause of the disease in view of the rapid restoration of 3-methylindole to normal levels by the second day after loading with L-tryptophan.

The pharmacokinetics of 3-methylindole (3MI) given orally in 2 doses (10 mg/kg and 20 mg/kg) to horses were determined. The pharmacokinetic plasma-concentration profiles for 3MI (10- and 20-mg/kg dosages) in horses were represented by a 2-compartment open model with first-order absorption, as determined by nonlinear least-squares regression analysis. Absorption of 3MI at both dosages was rapid. Comparisons of the peak plasma concentrations, the postdistribution half lives, total clearances, and areas under the curve of the plasma-concentration profiles between the 10- and the 20-mg/kg dosages may indicate the presence of nonlinear or dose-dependent kinetics for 3MI in horses.

Ruminal administration of 3-methylindole in goats severe pulmonary edema and respiratory distress. Electron microscopic studies of lungs reveal extensive degeneration and necrosis of alveolar membranous pneumocytes and bronchiolar epithelium. The necrosis of the pneumocytes is followed by proliferation of granular pneumocytes, which repopulate the alveolar basal lamina scaffold. 3-Methylindole may also induce proliferation of smooth endoplasmic reticulum in the remaining membranous pneumocytes and nonciliated columnar cells, indicating that these two cell types are involved in the xenobiotic function of the lung. The results suggest that 3-methylindole in cigarette smoke may play an important role in the pathogenesis of small airway disease and emphysema, and that patients with severe liver diseases or portocaval shunt may be predisposed to diffuse alveolar damage by 3-methylindole produced in the intestinal tract.