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Genetic resistance to gastrointestinal worms is a complex trait of great importance in both livestock and humans. In order to gain insights into the genetic architecture of this trait, a mixed breed population of sheep was artificially infected with Trichostrongylus colubriformis (n=3326) and then Haemonchus contortus (n=2669) to measure faecal worm egg count (WEC). The population was genotyped with the Illumina OvineSNP50 BeadChip and 48 640 single nucleotide polymorphism (SNP) markers passed the quality controls. An independent population of 316 sires of mixed breeds with accurate estimated breeding values for WEC were genotyped for the same SNP to assess the results obtained from the first population. We used principal components from the genomic relationship matrix among genotyped individuals to account for population stratification, and a novel approach to directly account for the sampling error associated with each SNP marker regression. The largest marker effects were estimated to explain an average of 0·48% (T. colubriformis) or 0·08% (H. contortus) of the phenotypic variance in WEC. These effects are small but consistent with results from other complex traits. We also demonstrated that methods which use all markers simultaneously can successfully predict genetic merit for resistance to worms, despite the small effects of individual markers. Correlations of genomic predictions with breeding values of the industry sires reached a maximum of 0·32. We estimate that effective across-breed predictions of genetic merit with multi-breed populations will require an average marker spacing of approximately 10 kbp.
Gastrointestinal nematode parasitism is the most important disease affecting livestock production systems in developing countries, particularly small ruminant production systems. Of particular importance are infections with the strongyle Haemonchus contortus. Integrated disease control strategies are required, including improved management, nutrition and wise use of anthelmintic chemicals. Increasingly, selection of sheep or goats for improved nematode resistance is viewed as a valuable option to complement other control measures. Breeding for resistance is possible because of the existence of extensive genetic variation in resistance, both within and between breeds of sheep and goats. Such breeding schemes are most likely to be based on choice of appropriate breeds adapted to the local environmental conditions, followed by phenotypic selection for resistance. Goal and selection objective traits are likely to include performance (e.g. growth rate) under conditions of parasite challenge, faecal egg count (FEC) and measures of anaemia. With current technologies, genetic markers are likely to be too expensive and logistically difficult to incorporate into breeding schemes in tropical or developing countries. Genotype by environment interactions may be expected, particularly when comparing animals in environments that differ in the extent of parasite challenge or differ in the quality of available nutrition. However, there is no reason to expect antagonistic genetic relationships between performance and resistance, and selection indices should be readily constructed that improve both performance and resistance. If FEC is decreased, then pasture contamination should also decrease, leading to additional benefits for all sheep grazing the same pasture. Finally, breeding for nematode resistance should lead to lasting and sustained improvements in resistance or tolerance. There is no empirical evidence to suggest that nematodes will evolve rapidly in response to resistant hosts, and mathematical models based on genetic and biological principles also suggest that resistance should be sustainable.
Twelve male 8-month-old lambs were used in a 6-month feeding experiment to determine the effects of dietary Mo, Mo + S and Zn supplements on the body retention and tissue accumulation of dietary Cu, Zn and Fe. The lambs were divided into four groups of three lambs each and each group was fed ad libitum one of four diets. A control diet was based on palm kernel cake (PKC) and grass hay. Three additional diets were the control supplemented with either Mo or Mo + S or Zn. At 3 months of the experiment, feces and urine were collected and sampled for 6 days. At the end of the experiment (6 months), blood was sampled and then the sheep were slaughtered. The liver and kidney were removed and sampled for chemical analysis. In comparison with the control, each dietary supplement decreased (P < 0.05) the Cu concentration in the liver, but only the Mo + S supplement decreased it to a safe range of below 350 μg/g dry matter. This was accompanied by the body retention of dietary Cu of 24.6%, 6.7%, 2.5% and 6.5% for the control, Mo, Mo + S and Zn treatments, respectively. The blood plasma concentration of Cu was decreased (P < 0.05) by the Zn supplement, but was not affected by other supplements (P > 0.05). It was concluded that from the supplements tested, only Mo + S appeared to be effective in reducing the retention and liver accumulation of the dietary Cu to prevent chronic Cu toxicity in sheep fed PKC-based diets.
Colostrum intake from birth to 24 h after the onset of parturition (T24) was estimated for 526 piglets from 40 litters. Plasma concentrations of immunoglobulin G (IgG), lactate, glucose and cortisol were determined at T24 for six piglets per litter. Plasma IgG concentration was also assayed at weaning (28 days) on the same piglets. Rectal temperature was measured at T24 on all piglets. Mortality was recorded until weaning and comparisons were made between piglets that died before weaning and those that were still alive at weaning. The piglets that died before weaning had lower birth weight, lower colostrum intake, lower weight gain between birth and T24, and had a lower rectal temperature, higher plasma cortisol concentration and lower plasma IgG and glucose concentrations at T24 than piglets still alive at weaning. In addition, a higher proportion of piglets that died before weaning had difficulty taking their first breath after birth and were affected by splayleg. Considering all piglets, colostrum intake was positively related to rectal temperature and plasma glucose concentration and negatively related to plasma cortisol concentration at T24. Plasma IgG concentration at T24 was explained by colostrum intake, IgG concentration in the ingested colostrum, birth weight and birth rank (P < 0.0001). Plasma IgG concentration at weaning was related to plasma IgG concentration at T24 (r = 0.54; P < 0.0001) and to colostrum intake (r = 0.32; P < 0.0001). Finally, body weight was explained by colostrum intake, birth weight and age until 6 weeks of age (P < 0.0001). These results show that colostrum intake is the main determinant of piglet survival through provision of energy and immune protection and has potential long-term effects on piglet growth and immunity.
The study aimed at comparing three strategies of supplementing selenium (Se) during the finishing period of Charolais young bulls: (1) administration of sodium selenite throughout the finishing (NaSe); (2) administration of an Se-enriched yeast strain (Saccharomyces cerevisiae NCYC R397) throughout the finishing (Se-Y); (3) administration of sodium selenite for 140 days replaced by Se-enriched yeast during the last 70 days of finishing (Switch). Eighty-four young bulls (mean initial BW = 434.2 ± 31.9 kg; mean age = 382 ± 52 days) were stratified by live weight and equally assigned to one of three Se treatments. Experimental groups were fed the same diets and the inclusion rate of the different treatments was targeted to achieve 0.3 mg of Se/kg of dry matter (DM) in the complete feed. The average daily gain of bulls was 1.36 kg/d and no differences due to Se treatment were recorded. Dry matter intake and feed conversion ratio were not affected by Se treatment resulting in, on average, 10.3 kg/d and 7.65, respectively. Repeated blood samples were taken at days 0, 120, 180 and 210 of finishing to assess the Se status of the animals. As compared to NaSe, both organic Se treatments (Se-Y and Switch) increased plasma Se in the last two sampling sessions according to a significant treatment × time interaction (P < 0.001). A similar trend was observed for serum total antioxidant status of the young bulls, whereas there was only a significant time effect (P < 0.001) on glutathione peroxidase activity that was raised by all Se treatments. The finishing period lasted 210 days and at the abattoir there were no differences across Se treatments in carcass weight and dressing percentage. A higher Se content in the Longissimus thoracis (LT) muscle was instead observed in Se-Y samples as compared with NaSe (0.85 v. 0.47 mg/kg DM; P < 0.05). Meat quality evaluation was carried out on LT samples after 6 and 11 days of ageing under a vacuum package. Regardless of ageing time, meat from young bulls supplemented with Se yeast had higher colour lightness (L*) values than those receiving NaSe (38.1 v. 36.6; P < 0.01) and showed a significant decrease in shear force (3.69 v. 4.22 kg/cm2; P < 0.01). The outcomes of the study suggest that the provision of Se yeast throughout the finishing period is a strategy to increase the benefits of the replacement of sodium selenite with organic selenium in beef cattle.
Colostrum provides newborn piglets with energy, immunoglobulins and growth, thereby playing an essential role in piglet survival. However, colostrum yield and composition are highly variable among sows. Some of the factors involved in this variability have been identified. The aim of the study was to confirm previous findings on a large number of animals and to investigate other potential factors of variation, such as the process of farrowing and the morphological changes of the mammary epithelium that occur during the 24 h postpartum. The experiment was conducted on 16 Large White (LW) and 56 Landrace (LR) × Large White (LR × LW) crossbred sows of mixed parities and their litters. Most farrowings were induced at 113 days of gestation and all farrowings were attended. Each piglet was weighed at birth and 24 h after farrowing started (t24). Colostrum ingestion by individual piglets was estimated using piglet weight gains from birth to t24. Colostrum production by sows was estimated by summing up colostrum intakes by each piglet of the litter. Colostrum was collected at the onset of farrowing (t0) and at t24 to determine concentrations of immunoglobulins G (IgG), Na and K. Analyses of correlations and multiple regressions were performed to identify the variables involved in variation of colostrum yield and IgG concentrations. Colostrum yield was not related to litter size and weight (P > 0.1). It was negatively correlated with the number of stillborn piglets (r = −0.33, P = 0.005) and within-litter variation of piglet birth weight (r = −0.24, P = 0.04). It was not related to the Na/K ratio in the colostrum, which is an indicator of the integrity of the mammary epithelium. When sows were categorised according to their level of colostrum yield, sows that produced a low yield of colostrum had more stillborn piglets at birth than the other sows (P < 0.05) and tended to have a longer birth interval during the early process of parturition (P < 0.1). At t24, concentrations of IgG in the colostrum were positively correlated with the Na/K ratio in the colostrum (r = 0.53, P < 0.001), which indicates the concomitance of the cessation of IgG transfer to the colostrum and the changes in the morphology of the mammary epithelium. This study points out the need for future research on the role of the hormones involved in both the process of parturition and lactogenesis in the relationship between stillbirth, process of parturition and colostrum production.
The objective of this study was to evaluate the effects of vegetable oil supplementation of ewe diets on the performance and fatty acid (FA) composition of their suckling lambs. Forty-eight pregnant Churra ewes (mean BW 64.3 ± 0.92 kg) with their 72 newborn lambs (prolificacy = 1.5) were assigned to one of four experimental diets, supplemented with 3% of hydrogenated palm (PALM), olive (OLI), soya (SOY) or linseed (LIN) oil. Lambs were nourished exclusively by suckling from their respective mothers. Ewes were milked once daily, and milk samples were taken once a week. When lambs reached 11 kg, they were slaughtered and samples were taken from musculus longissimus dorsi (intramuscular fat) and subcutaneous fat tissue. No changes were observed in milk yield, proximal composition or lamb performance (P > 0.10). Milk and lamb subcutaneous and intramuscular fat samples from the PALM diet had the highest saturated fatty acid concentration, whereas those of the OLI, SOY and LIN diets had the lowest (P < 0.05). The greatest monounsaturated fatty acid concentration was observed in milk from ewes fed OLI, and the least in milk and in lamb subcutaneous and intramuscular fat samples from LIN and PALM diets. Milk and lamb fat from ewes fed PALM displayed the highest 16:0 proportion and the lowest 18:0 (P < 0.05). There were higher concentrations of cis-9 18:1 in OLI samples (P < 0.05), more 18:2n-6 in SOY lambs and milk fat (P < 0.001) and the highest levels of 18:3n-3 and 20:5n-3 in LIN samples (P < 0.01). Milk and lamb subcutaneous and intramuscular samples from SOY and LIN diets contained the most cis-9, trans-11 conjugated linoleic acid, whereas PALM samples had the least (P < 0.01). Sheep diet supplementation with different oils, constituting up to 3% of their diets, resulted in changes in the FA composition of milk and the subcutaneous and intramuscular fat of suckling lambs, but did not affect either milk production or lamb performance.
Crow et al. (1990) and Barton (1992) have examined the critical migration rate for swamping selection in the nuclear system. Here, I use the same methodology to examine the critical migration rate in the cytonuclear system for hermaphrodite plants with a mixed mating system. Two selection schemes for a nuclear gene (heterozygote disadvantage and directional selection) and the directional selection scheme for organelle genes are considered. Results show that under random mating, the previous results are applicable to plant species by appropriate re-parameterization of the migration rate for nuclear and paternal organelle genes. A simple complementary relationship exists between seed and pollen flow in contributing to the critical migration rate. Under the mixed mating system, the critical migration rate of seeds and pollen for nuclear and paternal organelle genes can be changed due to the effects of selection and the cytonuclear linkage disequilibrium generated by migration and inbreeding. A negative but not complementary relationship exists between seed and pollen flow in contributing to the critical migration rate, varying with the mating system. Partial selfing can also adjust the critical seed flow for the maternal organelle gene, with a small critical migration rate for species of a high selfing rate. Both concordance and discordance among cytonuclear genes can occur under certain conditions during the process of swamping selection. This theory predicts the presence of various contributions of seed versus pollen flow to genetic swamping for plants with diverse mating systems.
The rate of insertion of transposable elements (TEs) is a fundamental parameter to understand both their dynamics and role in the evolution of the eukaryotic genome. Nonetheless, direct estimates of insertion rates are scarce because transposition is in general a rare phenomenon. A great deal of our previous work on transposition was based on a set of long-term mutation accumulation (MA) lines of Drosophila melanogaster started in 1987 (Oviedo lines), where roo was found highly active, with a rate of insertion of 7×10−4 insertions per element and generation, as compared with other 15 TE families that presented transposition rates around 10−5. Here, we study the evolution of the roo transposition rate, by in situ hybridization, after 60–75 additional generations of MA in two subsets of the Oviedo lines, O and O′, which had achieved average numbers of roo insertions of 77 and 84, respectively. In the O lines, insertions accumulated at a rate that remained constant (7×10−4 insertions per element and generation); however, the subset of lines O′ showed a lower accumulation rate of 4×10−4 insertions per element per generation, suggesting a regulation of transposition that depends on the number of elements. However, one of the O′ lines reached a number of 103 insertions, departing from the group mean by 4·6 sd, and showing that it escapes regulation. Hence, ‘de novo’ mutations affecting the regulation of transposition are relatively common. These results are discussed in relation to the possible mechanisms of containment of TEs.
Pannon White (n = 12) male rabbits (weight: 4050 to 4500 g, age: 9 months) received 2 ml of a suspension containing purified T-2 toxin by gavage for 3 days. The daily toxin intake was 4 mg/animal (0.78 to 0.99 mg/kg body weight (BW)). Control animals (n = 12) received toxin-free suspension for 3 days. Since a feed-refusal effect was observed on the second day after T-2 administration, a group of bucks (n = 10) were kept as controls (no toxin treatment) but on a restricted feeding schedule, that is, the same amount of feed was provided to them as was consumed by the exposed animals. On day 51 of the experiment (i.e. 48 days after the 3-day toxin treatment), semen was collected, and pH, concentration, motility and morphology of the spermatozoa, as well as concentration of citric acid, zinc and fructose in the seminal plasma, were measured. After gonadotropin-releasing hormone (GnRH) analogue treatment, the testosterone level was examined. One day of T-2 toxin treatment dramatically decreased voluntary feed intake (by 27% compared to control, P < 0.05) and remained lower (P < 0.05) during the first 2 weeks after the withdrawal of the toxin. BW of the contaminated rabbits decreased by 88% on days 17 and 29 compared to controls (P < 0.05). No effect of toxin treatment was detected on pH and quantity of the semen or concentration of spermatozoa. The ratio of spermatozoa showing progressive forward motility decreased from 65% to 53% in the semen samples of toxin-treated animals compared to controls (P > 0.05). The ratio of spermatozoa with abnormal morphology increased (P < 0.05) in the ejaculates collected from the toxin-treated animals. T-2 toxin applied in high doses decreased the concentration of citric acid in seminal plasma (P < 0.05). No effect of T-2 toxin on the concentrations of the other seminal plasma parameters (fructose and zinc) was observed. T-2 toxin decreased the basic testosterone level by 45% compared to control (P < 0.01) and resulted in lower (P < 0.05) GnRH-induced testosterone concentration. Feed restriction, that is, less nutrient intake, resulted in more morphologically abnormal spermatozoa in the semen, but it did not cause significant loss in BW, motility of the spermatozoa, composition of the seminal plasma or testosterone concentration – its effect needs further examination.
Two experiments were conducted to investigate the requirement for dietary crude protein (CP) in growing blue-breasted quail (BBQ). In Experiment 1, 300 1-day-old quails were randomly assigned to 10 groups according to a 2 × 5 factorial arrangement of treatments with two metabolisable energy (ME) levels (12.13 and 13.39 MJ/kg) and five CP concentrations (160, 190, 220, 250 and 280 g/kg) for 8 weeks. In Experiment 2, 300 1-day-old quails were subjected to a different factorial arrangement of treatments with two ME levels (11.51 and 12.13 MJ/kg) and five CP concentrations (210, 220, 230, 240 and 250 g/kg) for 28 days. Experiment 1 revealed that an interaction existed in weight gain between ME and CP levels in weeks 1 to 4. In both ME groups, quails receiving CP of 160 g/kg showed the least weight gains (P < 0.05). No differences (P > 0.05) existed in weight gain between the ME groups in which quails ingested CP of 250 and 280 g/kg, whereas quails consuming CP of 220 g/kg with an ME of 13.39 MJ/kg had smaller weight gain than did those ingesting higher CP concentrations (P < 0.05). Of main effects for weeks 1–4, quails treated with an ME of 12.13 MJ/kg consumed more feed than did those receiving another ME level, whereas quails in both ME treatments showed similar feed efficiencies. For weeks 5 to 8, no difference (P > 0.05) in weight gain, feed intake and feed efficiency was seen regardless of ME levels, and no interaction existed between ME and CP levels. In Experiment 2, the best weight gain and feed efficiency were achieved when the dietary CP concentration was more than 210 g/kg, and quails treated with 11.51 MJ/kg showed better weight gain and feed efficiency (P < 0.05) than did those that received 12.13 MJ/kg. Furthermore, the weight gains and protein intakes on the basis of per MJ from the two experiments were pooled together to estimate the protein intake necessary for the best growth performance by two mathematic models; they were then converted to dietary CP concentrations of 204 (minimum) and 233 g/kg (maximum) when ME was 11.51 MJ/kg. In conclusion, BBQ will achieve good growth performance with dietary CP of more than 204 g/kg on the basis of an ME of 11.51 MJ/kg in weeks 1 to 4.
The objectives of this study were to investigate the effects of different dietary sources of unsaturated fatty acids (fish oil (FO) and/or linseed oil (LO)) on laying performance, egg yolk fatty acid composition, ovarian follicular development, antioxidative properties, immune response and tibial bone characteristics in aged laying hens. A total of 100 Hisex Brown hens at 56 weeks of age were housed individually in laying cages in an open-sided building under a 16 h light:8 h dark lighting schedule. Hens were randomly divided into four experimental treatments (n = 25 each). Birds were fed ad libitum diets containing 2.5% vegetable oil (C, control), 2.5% FO, 2.5% LO and a mixture of 1.25% LO +1.25% FO (LO + FO) from 56 to 68 weeks of age. Egg production, egg quality characteristics and yolk lipid profile were analyzed. At 68 weeks of age, ovarian follicles were classified and tibial bone characteristics were determined. Serum thiobarbituric acid reactive substance (TBARS), glutathione peroxidase (GSH-Px) activity and total antioxidant capacity were measured. Incorporation of n-3 polyunsaturated fatty acids (n-3PUFA) into the egg yolks has been successful by using dietary FO and/or LO. There were no significant effects of treatments on hen-day egg production, feed intake, egg weight, egg shape index, albumen height, Haugh units and yolk height. However, dietary FO and/or LO supplementation had a significantly positive effect on eggshell percentage, eggshell thickness and yolk color. At 68 weeks of age, there was no significant difference among dietary treatments for tibial bone measurements. Also, no negative effects were detected in ovarian follicular development and weights of the ovary and oviduct, expressed in both absolute terms and relative to body weight. Dietary 2.5% LO, 2.5% FO and a mixture of 1.25% FO + 1.25% LO enhanced GSH-Px activity, total antioxidant capacity and antibody titers significantly in comparison with control. It could be concluded that inclusion of mixed sources of n-3PUFA in diets at moderate levels (2.5%) increased the n-3PUFA content and decreased the n-6/n-3 ratio content in the yolk, improved the antioxidative status, reduced lipid peroxidation, enhanced the antibody response and did not have any negative influence on ovarian follicular development and tibial bone characteristics in aged laying hens.
The aim of this study was to investigate protein requirements for the maintenance and growth of blue-breasted quail (Excalfactoria chinensis) from 7 to 21 days of age. A total of 180 quails, 7 days old, were randomly assigned to 36 cages and for 2 weeks were fed diets with a metabolisable energy concentration of 12.13 MJ/kg and a dietary CP concentration of 125, 150, 175, 200, 225 or 250 g/kg. The average BW per cage and the feed intake per cage were recorded daily. The results showed that quails fed 125 g/kg CP could not maintain their BW and had negative feed efficiency. There were linear and quadratic relationships between CP level and response criteria, including BW, weight gain, feed intake, feed efficiency, final body nitrogen mass and body nitrogen accretion (P < 0.05). The dietary CP requirements, as calculated using a one-slope quadratic broken-line model, were 211 and 202 g/kg according to weight gain and feed efficiency, respectively. The regression equations, on the basis of metabolic BW, of daily weight gain on daily protein intake according to the model were Y = 0.137−2.128(0.113−X) if X < 0.113 and Y = 0.137 if X ⩾ 0.113 (R2 = 0.96, P < 0.001), which meant that the protein requirement for maintenance was 0.049 times the metabolic BW and that to gain 1 g weight quails needed to ingest an extra 0.47 g protein after the maintenance requirement was satisfied. The regression equations, on the basis of metabolic BW, of daily body nitrogen accretion on daily protein intake according to the model were Y = 5.667−76.700(0.119−X) if X < 0.119 and Y = 5.667 if X ⩾ 0.119 (R2 = 0.95, P < 0.001), which meant that quails had to receive an amount of protein equal to their metabolic BW multiplied by 0.045 to satisfy the requirement for maintenance and then ingest an extra 13 g protein to accrete 1 g body nitrogen. In conclusion, growth or protein accretion rates should be regulated according to dietary CP for specific experimental purposes via apportioning protein requirements for maintenance v. growth.
Intensively finishing cattle on a high-grain diet is generally used to enhance marbling, whereas extensively finishing on grass is known to provide improved muscle fatty acid profiles. The objective of this study was to evaluate to what extent intensive concentrate finishing (0, 1 or 2 months) can be combined with forage feeding without negatively affecting the fatty acid profile of genetically lean animals. Bulls from the ‘Asturiana de los Valles’ breed were reared under grazing conditions with/without final finishing on a barley-based concentrate: 0 months (control; n = 7), 1 month (n = 10) and 2 months (n = 7). Yearling bulls were slaughtered commercially at an average live weight of 516 ± 9.8 kg. Increasing the finishing time on concentrate significantly increased the saturated and monounsaturated fatty acids, whereas polyunsaturated fatty acids (PUFAs) tended to decrease and it was not possible to increase the long-chain PUFA content in muscle tissue of this breed. An increase was observed for total trans-18:1 (average 5.5% with grain v. 3.7% for grass). The 11t-18:1/10t-18:1 ratio was significantly higher in grass-fed (average 8.1) compared with grain-finished animals (average 1.1). Grass or limited concentrate finishing reduced the n-6/n-3 ratio in muscle tissue (average 3.6 for 0 and 1 month, and 4.9 for 2 months on grain finishing). The beef was within or close to the recommended values for human consumption (i.e. polyunsaturated/saturated > 0.45, n-6/n-3 < 4.0), and total trans-FA content was low. However, finishing increased the content of undesirable trans-18:1 and conjugated linoleic acid isomers, particularly after 2 months, whereas grass finishing was judged to provide a healthier beef fatty acid profile.
Today, different analytical methods are used by different laboratories to quantify androstenone in fat tissue. This study shows the comparison of methods used routinely in different laboratories for androstenone quantification: Time-resolved fluoroimmunoassay in Norwegian School of Veterinary Science (NSVS; Norway), gas chromatography coupled to mass spectrometry in Co-operative Central Laboratory (CCL; The Netherlands) and in Institut de Recerca i Tecnologia Agroalimentàries (IRTA; Spain), and high-pressure liquid chromatography in Agroscope Liebefeld-Posieux Research Station (ALP; Switzerland). In a first trial, a set of adipose tissue (AT) samples from 53 entire males was sent to CCL, IRTA and NSVS for determination of androstenone concentration. The average androstenone concentration (s.d.) was 2.47 (2.10) μg/g at NSVS, 1.31 (0.98) μg/g at CCL and 0.62 (0.52) μg/g at IRTA. Despite the large differences in absolute values, inter-laboratory correlations were high, ranging from 0.82 to 0.92. A closer look showed differences in the preparation step. Indeed, different matrices were used for the analysis: pure fat at NSVS, melted fat at CCL and AT at IRTA. A second trial was organised in order to circumvent the differences in sample preparation. Back fat samples from 10 entire males were lyophilised at the ALP labortary in Switzerland and were sent to the other laboratories for androstenone concentration measurement. The average concentration (s.d.) of androstenone in the freeze-dried AT samples was 0.87 (0.52), 1.03 (0.55), 0.84 (0.46) and 0.99 (0.67) μg/g at NSVS, CCL, IRTA and ALP, respectively, and the pairwise correlations between laboratories ranged from 0.92 to 0.97. Thus, this study shows the influence of the different sample preparation protocols, leading to major differences in the results, although still allowing high inter-laboratory correlations. The results further highlight the need for method standardisation and inter-laboratory ring tests for the determination of androstenone. This standardisation is especially relevant when deriving thresholds of consumer acceptance, whereas the ranking of animals for breeding purposes will be less affected due to the high correlations between methods.